JPH0356460A - Proline derivative - Google Patents

Abstract

NEW MATERIAL:Compounds of formula I (X is H or OH; R is five-membered saturated heterocycle which may contain S) and salts thereof. EXAMPLE:(S)-N-[(R)-(-)-Thiazolidine-4-carbonyl]-proline benzyl ester. USE:A safe and hypotoxic remedy for amnesia having a prolyl-end peptidase inhibition activity. PREPARATION:Either a carboxylic acid of formula R-COOH or a reactive and functional derivative (e.g. acid halide or acid anhydride) thereof is reacted with a compound of formula II to obtain a compound of formula I. The reaction is carried out in the presence of a condensation agent (e.g. N,N'- dicyclohexylcarbodimide) and a base (e.g. triethylamine).

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to proline derivatives useful as pharmaceuticals.

More specifically, the present invention relates to prolyl endopeptidase (Prolyl Bndopeptidase).
General formula (in the formula, The present invention provides a proline derivative represented by the following cyclic group and a pharmacologically acceptable salt thereof.

[Conventional technology]

As the population ages, issues of geriatric medical care are becoming more important. Among these, senile dementia is a serious social problem, so there is a desire for the prompt development of effective therapeutic agents.

So far, as therapeutic agents for amnesia, dementia, and the like, agents for improving cerebral circulation through cerebral vasodilation and the like, and agents for activating cerebral metabolism through activation of brain cell oxygen consumption, etc., have been used. However, although these drugs are effective for dementia caused by cerebrovascular disorders, the drawback is that they are not certain to be effective against dementia caused by other causes.

PEP is known as an enzyme that acts on bioactive peptides and synthetic substrates containing proline and specifically cleaves the carboxyl side of proline. This enzyme is known as vasopressin, which is said to be associated with memory.
) and thyrotropin-releasing hormone (Thyrotrop
in Releasing }Iormone, TR
Various studies have been conducted on the relationship between the inhibitory activity of this enzyme and its anti-amnestic effect, and the results suggest that PEP inhibitors can be used as therapeutic agents for dementia and amnesia ( Biochemistry, Vol. 55, No. 8, Page 831, 1983).

[Problem to be solved by the invention]

Since the cerebral circulation improving agents and cerebral metabolism activating agents conventionally used as therapeutic agents for amnesia and dementia are not very effective, there has been a desire to develop amnesic therapeutic agents with new effects.

The present inventors conducted an investigation to find a therapeutic agent for amnesia that has an action different from that of conventional therapeutic agents, and found that certain proline derivatives exhibit strong PEP inhibitory activity and that the objective can be achieved.

The present invention is based on these findings.

[Means to solve the problem]

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

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

The compound of the general formula (1) of the present invention can be produced as follows. For example, a carboxylic acid represented by the general formula R-COOH (n) (R in the formula has the same meaning as above) or a reactive functional derivative thereof, and a carboxylic acid represented by the general formula (X in the formula has the same meaning as above), It can be produced by reacting a compound represented by

Here, in the compound represented by general formula (n), if it is necessary to protect the amine 7 group, protect the amino group with a suitable amino protecting group, for example, t-butoxycarbonyl group, according to a conventional method. After that, reaction is performed, and then the amino protecting group is removed to obtain the target compound (1).

In the production method of the present invention, the compounds of general formulas (II) and (II[) used as starting materials are available as commercial products or can be easily produced by methods described in literature.

When producing the compound of the general formula (I) of the present invention, when the carboxylic acid of the one-shell formula (It) is reacted with the compound represented by the general formula (I[), in the presence of a condensing agent and a base. The condensing agent used at this time is a condensing agent commonly used in peptide aggregation, such as N, N'-dicyclohexylcarbodiimide, N-ethoxycarbonyl-2-ethoxy-1,2- Examples include dihydroquinoline, and examples of the base include triethylamine.

In the method for producing the compound of general formula (I) of the present invention, when a reactive functional derivative of a carboxylic acid represented by general formula (I[) and a compound represented by general formula (I) are reacted, , the reaction is carried out in the presence of a base, and the reactive functional derivatives of the compound of general formula (II) include acid halides,
Acid anhydrides, mixed acid anhydrides, active esters, etc. can be mentioned, and the bases used at this time include pyridine,
Bases such as triethylamine can be mentioned.

In order to suitably carry out the method for producing the compound of the general formula (I) of the present invention, for example, the carboxylic acid represented by the general formula (II) or, if necessary, its N-protected form and an equimolar amount thereof. A compound represented by the general formula (nl) is dissolved in an inert organic solvent such as methylene chloride, ethanol, etc., a necessary amount of base and a condensing agent are added, and the mixture is heated under ice-cooling to room temperature for 10 minutes.
Stir for ~20 hours, process and purify according to conventional methods to obtain the desired product.

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

The compound of formula (I) of the present invention contains a proline moiety of 1
Although it has ~2 asymmetric carbon atoms, in the present invention, the substituent on each asymmetric carbon may be either R or S, or a mixture thereof. Each optically active compound uses an optically active compound as a starting material,
It can be obtained by steric condensation.

The compound of general formula (I) of the present invention can be made into various pharmaceutical preparations according to conventional methods. That is, by adding pharmaceutical additives such as excipients, disintegrants, condensing agents, and lubricants as necessary, and preparing according to conventional methods, various preparations such as tablets, powders, granules, capsules, etc. can be prepared. It can be done.

When the compound of general formula (1) of the present invention is used as a therapeutic agent for amnesia, the dosage is appropriately determined depending on the patient's age, weight, sex, degree of symptoms, etc.; For administration, it is used in the range of 50 to 1000 mgs, and in the case of parenteral administration, it is used in the range of 1 to 500 mgs.

〔Effect of the invention〕

The compound of the general formula (I) of the present invention is N-carbobenzooxy-glycyl-L-brolyl-β-naphthylamide (hereinafter referred to as Z-Gly-Pro-β-NA).
In an inhibitory activity measurement test against bovine brain-derived prolyl endopeptidase using the substrate, approximately 1 x 10 to 7
It shows 50% inhibitory activity at IO-' molar concentration.

Preferably, it is (S) -N- {(R) - (-) monothiazolidine-4-carbonyl}proline pendyl ester hydrochloride, and its IC5. value is? ．． 2X10-
'It's a mole.

As described above, the compound of the general formula (I) of the present invention exhibits strong PEP inhibitory activity and has low toxicity, so it is a safe and useful compound as an excellent amnesia therapeutic agent.

〔Example〕

The following Reference Examples and Examples are given to explain the present invention in more detail. Note that the melting points of the compounds in each Reference Example and Examples are all uncorrected.

Reference Example 1 (R)-(-)monothiazolidine-4-carboxylic acid 13.
3 g and triethylamine l4- in dioxane 50 m
Dissolved in a mixed solvent of 1 and 50 mj' of water, added 24 g of Di-1-Ptyrouge carbonate under water cooling, and dissolved at room temperature.
Stirred for 0 hours. Add 100 ml of water to the reaction solution! Add. Wash with ethyl acetate and cool with water until the aqueous layer reaches pH 2.
A 0% aqueous citric acid solution was added. Extraction was performed with ethyl acetate, and the ethyl acetate layer was washed with saturated brine and dried over anhydrous magnesium sulfate. Distill the solvent under reduced pressure to obtain 21.6 g of the target product.
(93%) was obtained.

IR (KBr): vco 1745.
1630 cmNMR (CDCI3) δ: 1.4g (s, 9M), 3JO (s. 2H),
4J5~4.95 (m, 3H), 10. 10
(br-s, 1}1) Reference Example 2 The following compound was produced in the same manner as in Reference Example 1, using hydroxy-proline instead of (R)-(-)monothiazolidine-4-carboxylic acid. did.

Hydroxy-(S)-Nt-butoxyluponylbroline IR (KBr): vco 1725. 167
0 cm-NMR (CDCI.) δ: 1.34. 1.39 (S. 8. 9H,
isomer), 1y80-1.95 (m. IH). 2
．． 05 ~2.20 (m, ltl), 3.20 ~3
．． 45 (m, 2H). 4.11 (t. IH).
4.24 (s. LH). 5.04 (s. LH).
12.47 (s.LH) Reference Example 3 Hydroxy-broline benzine ester/hydrochloride 2.0 g of hydroxy-proline was dissolved in N,N-dimethylformamide IO-, and benzyl bromide 1.
14mj2 and 0.81 g of sodium hydrogen carbonate were added and stirred at 40°C for 14 hours. The reaction solution was distilled off under reduced pressure, and ethyl acetate was added to prepare a saturated aqueous sodium hydrogen carbonate solution.
It was washed successively with an aqueous citric acid solution, water, and saturated brine, and dried over anhydrous magnesium sulfate.

The ethyl acetate layer was distilled off under reduced pressure to obtain 2.4 g of hydroxy-(S)-Nt-phthoxycarbonyl-proline pendyl ester.

NMR (CDCI3) δ: 1.34. 1.45 (s.s. 9H,
isomer), 2.00-2.40 (m. 2H). 3
．． 40-3,65 (m, 2}1), 4. 35 ~4
．． 55 (m. 2H), 5.00 ~ 5.30
(m, 28), 7. 20 ~7. 40 (m,
5H) The above benzyl ester 2. 38 g was dissolved in 20 parts of ethyl acetate, hydrogen chloride gas was blown into the solution for 10 minutes under water cooling, and the mixture was stirred at room temperature for 1 hour. The reaction solution was distilled off under reduced pressure to obtain 5 g of IJ.

IR (KBr): L'CO 1745 c
m-'NMR (DMSO) δ: 1.95 to 2.30 (m, 2H). 3.0
8 (d. IH), 3.2 (1-3.50 (m. IH)
). 4.42 (s. IH), 4.45 (dd.
IH), 5.24 (dd. 2H). 5.58 (b
r-s. Iff), 7.30-7.55 (m, 5
H). 8.90-9.70 (br. IH). 9.
80-10.70 (br, 1}1) Example l L-7” loline benzyl ester hydrochloride 0.47 g,
(R)-(-)-Nt-butoxycarbonylthiazolidine-4-carboxylic acid 0. 44 g and 0.28 g of triethylamine were added to 5 g of dry methylene chloride, and the mixture was stirred under water cooling. Furthermore, 1-hydroxybenzotriazole l hydrate 0. 46 g and 0.45 g of N,N"-dicyclohexylcarbodiimide were added, and the mixture was stirred at room temperature for 14 hours. After the reaction was completed, ethyl acetate was added and cooled with ice. The precipitated crystals were filtered off, and the mixture was stirred under reduced pressure. The solvent was distilled off.

Ethyl acetate was added to the residue, which was washed successively with a 10% aqueous citric acid solution, a saturated aqueous sodium bicarbonate solution, water, and saturated brine, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The residue was purified by silica gel column chromatography (elution solvent: chloroform) to give a 0. 4
9 g of (S)-N-((S)-Nt-butoxycarbonyl-thiaso'lysine-4-carbonyl}proline benzyl ester was obtained.

NMR (CDCI3) δ: 1.45 (S, 9ft), 1.80-2.3
0 (m, 4H). 3. 05-3. 35 (m.
2H), 3. 55 ~3. 90 (m,2}1
). 4.55-5.00 (m. 4H). 5.14
(dd,2}1). 7.20 ~7. 40 (m
．． 5}1) The above benzyl ester 0. 49 g of the solution was dissolved in 50 parts of ethyl acetate, hydrogen chloride gas was blown into the solution under water cooling until it reached saturation, and the mixture was stirred at room temperature for 5 hours. After the reaction, the solvent was distilled off under reduced pressure and recrystallized with a mixed solvent of methanol, methylene chloride and ether to give 0.
27 g of the target product was obtained.

IR (KBr): νco1740. 165
5 cm-MS: MH" 321 NMR (DMSO) δ: 1.80-2.05 (m, 2H). 2.10
~2J5 (m, IH). 2.80 (dd, IH).
3.20-3.80 (m, 4H). 4.28 (dd
, 2H), 4.49 (dd, LH). 4.67 (
t, LH). 5.13(d, 2H). 7.30
~7.45 (m, 5H), 8.20~11.10 (
br. 2N)\Example 2 (S)-Nt-
The following compound was produced in the same manner as in Example 1 using butoxyluponylproline.

IR (KBr): I”Go 1745.
1650 cm-'MS: MH"
303NMR (DMSG) δ: 1.65-2.70 (m, 8H). 3
．． 20-3.90 (m, 4H). 4.50 ~4,
70 (m, 2H). 5. 20 ~5. 3
0 (ill. 2H), 7.40-7.65 (m
, 511), 8.60-10.00 (br,
2}1) Example 3 The following compound was produced in the same manner as in Example 1, using hydroxyl-proline benzyl ester/hydrochloride instead of L-proline benzyl ester/hydrochloride.

IR (KBr): l'co 1740.
1650 cm-'NMR (DMSO) δ: 1,90-2.05 (m. 1}1). ．． 2.
15~2JO (m, LH), 2.81 (dd,
11{), 3.50-3.75 (m, 3H),
4.28 (dd, 211), 4.40 (s,
Ift), 4.51 (t. IH). 4.
77 (t, IH). 5.13 (d. 2}1
). 7JO ~7.45 (m. 5H). 9.2
0 to 11.10 (br-s, 2H) Elemental analysis value' (as Cl8H21N204SC1) C% H% N% Calculated value 51.53 5.67 7.51 Actual value 50.86 5.71 7.51 Example 4 L-florin benzyl ester salt [salt 1.0 g, pyroglutamic acid 0.52 g and triethylamine 0.5
Dissolve 8 g in ethanol 14-, N-ethoxycarbonyl-2-ethoxy-1,2-dino\hydroquinoline 0
．． 99 g was added and stirred at room temperature for 20 hours. The solvent was distilled off under reduced pressure, ethyl acetate was added, and insoluble materials were filtered off. The filtrate was washed with a saturated aqueous sodium bicarbonate solution and water, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography (elution solvent: chloroform/ethanol = 30/
1') to yield 192 mg of the target product (diastereomer A, 73; diastereomer B, 119
mg) was obtained.

Diastereomer A Rf value: 0.53 (Developing solvent: chloroform/
Ethanol = 7/1) IR (KBr): L'CO 1740, 1
695. 1650 cm-'MS: MH"
317 NMR (CDCI3) δ: l. 85-2.60 (m, IE). 3.45
~3,80(m,2}1). 4. 40 ~4. 6
0 (m. 2tl). 5. 17 (dd, 2H)
, 5.90 (s. IH), 7JO ~7.45 (
m. 5H) Diastereomer B Rf value: 0.58 (Developing solvent: chloroform/
Ethanol = 7/1) IR (κBr): vco 1735.
1640 cmMS: MH" 3
17 NMR (CDCI3) δ: 1.90 to 2.50 (m, 8}1).
3.57 (t, 2H), 4.30 ~ 4.40 (
m, ltl), 4.61 (dd. lft)
, 5.15 (dd, 2H). 6.16(s,
IH), 7.30 ~ 7.45 (+++. 5
N) Example 5 PEP inhibitory activity measurement experiment 2 - Using Gly-Pro-β-N^ as a substrate, the inhibitory activity against bovine brain-derived PEP was measured.

(Measurement method) 20 mM Lis-HCl buffer containing 10 mM BDTA and 10 mM 2-mercaftethanol (20 d
-Tris HCI Buffer, pH=7.0)
PPP in 0.7d (approximately 0.14 u/rnl)10
0 d and each concentration (0, 10-9 to 10-' M)
Add 100 d of a solution of the test compound adjusted to
Preincubate for 5 minutes at °C.
ation) Shita.

Then, to this, each concentration (5.0, 2.5, 1.25, 0.625
, 0.3125 mM) was added and incubated again at 37°C for 1
Incubation was performed for 5 minutes to allow the enzymatic reaction to proceed. The reaction was stopped with 25% trichloroacetic acid and
r. p. Centrifuge for 10 minutes at 0.5 m,
0.5 ml of 0.1% nitrous acid was added thereto, and after 3 minutes, a 0.05% ethanol solution of N-(1-naphthyl)ethylenediamine dihydrochloride was added. After leaving the mixture at 37°C for 25 minutes, 570
Measure the absorbance at nm, calculate the oxygen activity at each concentration using the following formula, and calculate the 50% inhibition concentration (
I.C. . value〉 was calculated.

Enzyme activity unit (μmo17min/11112)
=ΔOn X0. 42X dilution rate (results) Compound Compound A Compound C IC,. Value 720 μM 1,0 mM

Claims (1)

[Claims] General formula▲ Numerical formula, chemical formula, table, etc.▼ (In the formula, A proline derivative represented by (which is a cyclic group) and a pharmacologically acceptable salt thereof.