JPH0826067B2 - Peptide derivative and anti-dementia agent containing the same - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a peptide having a nootropic effect and therefore useful as a medicine, particularly an anti-dementia agent, and a peptide derivative.

[PRIOR ART] Vasopressin has been known for a long time as having a cognitive effect, but a peptide which can be considered as a fragment of vasopressin recently, for example, Also, It has been reported that the peptide represented by [6] has a nootropic effect similar to vasopressin [Science 2
21 , 1310-1312 (1983)] [Brain Research (Brain R
eseartch) 371 , 17 (1986)].

[Problems to be Solved by the Invention] It is an object of the present invention to provide a novel peptide and a peptide derivative having a nootropic effect superior to those of vasopressin and vasopressin fragment peptides. is there.

[Means for Solving Problems] The present invention provides the following general formula (I): [Wherein A is cyclopentylcarbonyl, Pro or pGl
u is, B is Gly or β-Ala, W is a hydrogen atom or the following general formula (II): (In the formula, Y ¹ is H or CO-T, and Y ² is OH or T (where T is the following general formula (III): (In the formula, R ¹ is a group selected from the group consisting of an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to 10 carbon atoms, and an alkylthio group having 1 to 6 carbon atoms) Or the following general formula (IV): (Wherein R ² is a group selected from the group consisting of a hydrogen atom, an alkylcarbonyl group having 2 to 7 carbon atoms, and an arylcarbonyl group having 7 to 10 carbon atoms), )) Is a group represented by] or a peptide derivative represented by the following general formula (V): [Wherein A and B are the same as defined above], a derivative in the functional group thereof, or a pharmacologically acceptable salt thereof.

Furthermore, the present invention provides an effective amount of the peptide derivative represented by the above general formula (I), a derivative in a functional group thereof, or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable carrier or The present invention relates to an anti-dementia drug containing a diluent.

The derivative in the functional group of the peptide derivative represented by the above general formula (I) means the following.

a) N-acyl derivatives derived from aliphatic carboxylic acids having 1 to 6 carbon atoms, b) amides or mono-alkyl or di-alkyl substituted amides having an alkyl group of 1 to 6 carbon atoms, And c) an ester derived from an alcohol having 1 to 18 carbon atoms, preferably an aliphatic alcohol having 1 to 6 carbon atoms.

Examples of the pharmacologically acceptable salt of the above peptide derivative or the derivative in the functional group thereof include an acid addition salt and a basic salt. Such acid addition salts include salts of inorganic acids (eg, hydrochloric acid, sulfuric acid, phosphoric acid) or organic acids (eg, acetic acid, propionic acid, citric acid, tartaric acid, malic acid, oxalic acid, methanesulfonic acid). Can be mentioned. Examples of the basic salt include a sodium salt, a potassium salt, a triethylamine salt and the like.

In the present specification, amino acids, peptides, protecting groups, solvents and the like are abbreviations commonly used in the art, or IU
The abbreviations adopted by the PAC-IUB naming committee are used. For example, the following abbreviations are used. Amino acids mean L-form.

β-Ala: β-alanine Arg: Arginine Cys: Cysteine Gly: Glycine pGlu: Pyroglutamic acid Pro: Proline Boc: t-Butoxycarbonyl Z: Benzyloxycarbonyl Mbs: p-Methoxybenzenesulfonyl MBzl: p-Methoxybenzyl Acm: Acetamide Methyl Scm: Carbomethoxysulfenyl OBzl: Benzyl ester OSu: N-Hydroxysuccinimide ester DCC: N, N'-Dicyclohexylcarbodiimide DCUrea: N, N'-Dicyclohexylurea HOBt: 1-Hydroxybenzotriazole Et ₃ N: Triethylamine NMM: N-Methylmorpholine TFA: Trifluoroacetic acid MSA: Methanesulfonic acid AcOEt: Ethyl acetate AcOH: Acetic acid THF: Tetrahydrofuran DMF: N, N-Dimethylformamide MeOH: Methanol The peptide derivative of the present invention is first usually used in peptide chemistry. Methods such as Schrder and Lbke
Author "The Peptides" Volume 1, Academic
Press, New York, USA (1965), Nobuo Izumiya et al. "Basics and experiments of peptide synthesis" Maruzen Co., Ltd. (1985) and other methods (liquid phase method and solid phase method) be able to.

As a condensation method for forming a peptide bond, an azide method, an acid chloride method, an acid anhydride method, a mixed acid anhydride method,
N, N'-dicyclohexylcarbodiimide method, N, N'-dicyclohexylcarbodiimide-additive method, active ester method, carbonyldiimidazole method, redox method, method using Woodward reagent K, and the like.

Before performing the condensation reaction, by a means known per se,
A carboxyl group or amino group that does not participate in the reaction may be protected, or a carboxyl group or amino group that participates in the reaction may be activated.

Examples of carboxyl protecting groups include, for example, methyl,
Ethyl, benzyl, p-nitrobenzyl, t-butyl,
Esters such as cyclohexyl may be mentioned.

Examples of the amino group-protecting group include a benzyloxycarbonyl group, a t-butoxycarbonyl group, an isobornyloxycarbonyl group and a 9-fluorenylmethoxycarbonyl group.

Examples of the guanidino group-protecting group include a nitro group and
Examples thereof include a benzyloxycarbonyl group, a tosyl group, a p-methoxybenzenesulfonyl group and a 4-methoxy-2,3,6-trimethylbenzenesulfonyl group.

Examples of the mercapto-protecting group include a trityl group, an acetamidomethyl group, a benzyl group, a p-methoxybenzyl group, and a 3-nitro-2-pyridinesulfenyl group.

Examples of activated carboxyl groups include corresponding acid anhydrides, azides, active esters [alcohols (eg, pentachlorophenol, 2,4-dinitrophenol, cyanomethyl alcohol, p-nitrophenol, N- Hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-
Hydroxyphthalimide, ester with 1-hydroxybenzotriazole)] and the like. Activated amino groups include, for example, the corresponding phosphoric amides.

The reaction is usually performed in a solvent, for example, a solvent such as chloroform, dichloromethane, ethyl acetate, N, N-dimethylformamide, dimethyl sulfoxide, pyridine, dioxane, tetrahydrofuran, water, methanol, or a mixture thereof. be able to.

The reaction can be carried out at a temperature generally ranging from about -30 ° C to about 50 ° C.

The protective group elimination reaction of the peptide of the present invention depends on the type of the protective group used, but it does not affect the peptide bond and it is necessary to remove the protective group.

Examples of the method of removing the protective group include, for example, hydrogen chloride, anhydrous hydrogen fluoride, methanesulfonic acid, trifluoromethanesulfonic acid, trifluoroacetic acid, or an acid treatment with a mixture thereof, or the like. Reduction in sodium in liquid ammonia, palladium carbon, and the like are also included. In the above deprotection reaction by acid treatment, it is effective to add a cation scavenger such as anisole, phenol or thioanisole.

After completion of the reaction, the peptide and peptide derivative of the present invention thus produced can be obtained by means of peptide separation means known per se, such as extraction, partitioning, reprecipitation, recrystallization, column chromatography and the like. it can.

Further, the peptide and peptide derivative of the present invention can be converted into a derivative in the functional group as described above or a pharmacologically acceptable salt thereof by a method known per se.

The peptide derivative of the present invention exhibits a strong nootropic action in a passive avoidance test in rats.

Examples of useful target disease names of the peptide derivative of the present invention include senile dementia (Alzheimer type dementia), cerebrovascular dementia, and Alzheimer's disease, Pick's disease, Huntington's chorea, Creutzfeldt-Jakob's disease, Parkinson's disease, Examples thereof include dementia based on cerebellar spinal degeneration, etc., and can be used for the prevention or treatment of these diseases.

The toxicity of the peptide derivative of the present invention is extremely low, and there is no death even at a dose far exceeding the pharmaceutically effective dose.

The peptide derivative of the present invention can be administered as a free form or as a derivative at a functional group thereof. Regarding the dose, whether it is the free form or a salt thereof, an appropriate amount of the free form is generally in the range of 0.1 ng to 1 mg / day.
Particularly, for parenteral administration and nasal administration, 0.1 ng to 100 μg / day is preferable, and for oral administration and rectal administration, parenteral administration is 10 to 10 μg / day.
It is preferable to administer 100 times. The peptide derivative of the present invention is mainly administered parenterally (eg, intravenous or subcutaneous injection, intracerebroventricular or intrathecal administration, nasal administration, rectal administration). Good.

Dosage forms include, for example, injections, suppositories, powders, nasal drops, pills, tablets and the like. Although the peptide derivative of the present invention can be stored as a solution of physiological saline,
Mannitol or sorbitol may be added to it to be dissolved as a freeze-dried ampoule.

 Examples will be shown below.

In each example, the developing solvent for thin layer chromatography was as follows, and TLC plate silica gel 60F ₂₅₄ manufactured by Merck was used.

R _f ¹ : chloroform-methanol-acetic acid-water (80: 20: 2.5: 5) lower layer R _f ² : chloroform-methanol-water (70: 30: 5) R _f ³ : n-butanol-acetic acid-water (2 Purification by high performance liquid chromatography was performed using a column: μBondapak C ₁₈ 1.9 × 15 cm mobile phase: A) 0.05% TFA, B) acetonitrile.

[Reference Example 1] H-Cys (Scm) -T · hydrochloride (where T is a group represented by the above general formula (III) in which R ^{1 in} the formula is a benzoyl group) (1) Boc- Cys (Acm) -T Boc-Cys (Acm) -OH 1.0g, S-benzoyl thiamine
CH ₂ Cl ₂ 50 m of 1.3g and 4-dimethylaminopyridine 20mg
A solution of DCC 0.78 g in CH ₂ Cl ₂ 5 ml was added dropwise to the solution under ice cooling.

After stirring for 30 minutes under ice cooling and 1 hour at room temperature, DCUr
ea was filtered off and washed with saturated aqueous sodium hydrogen carbonate solution and water.

After drying over anhydrous sodium sulfate, the solvent was evaporated and the crystals were collected by filtration with ether to give the title compound.

Yield: 1.8 g Melting point: 71-75 ° C. R _f ¹ : 0.74 R _f ² : 0.82 [α] _D : −39.2 ° (c = 0.5, DMF) (2) Boc-Cys (Scm) -T Boc-Cys ( Acm) of _{-T 600mg MeOH-CH 2 Cl 2} (1: 1v / v)
Cl-Scm 0.14 ml was added to the 6 ml solution, and the mixture was stirred at room temperature for 20 minutes.

After the solvent was distilled off, the residue was purified by silica gel column with CHCl ₃ -MeOH to obtain the title compound as an oily substance.

Yield: 580 mg R _f ¹ : 0.82 R _f ² : 0.88 [α] _D : -44.0 ° (c = 0.5, DMF) (3) H-Cys (Scm) -T ・ hydrochloride Boc-Cys (Scm) -T 470 mg of 3 in 4 ml of 4N HCl-AcOEt.
After standing at room temperature for 0 minutes, the solvent was distilled off. The residue is converted into CHCl ₃ −
Purification on silica gel column with MeOH gave the title compound as an oil.

Yield: 250 mg R _f ¹ : 0.38 R _f ² : 0.54 [α] _D : + 38.4 ° (c = 0.5, DMF) [Example 1] (1) Z-Arg (Mbs) -Gly-NH ₂ Z-Arg (Mbs) -OH dicyclohexylamine salt 10 g
Was dissolved by stirring in 100 ml of AcOEt + 70 ml of 5% citric acid water, and then the AcOEt layer was washed with water and dried over anhydrous sodium sulfate.

The solvent was distilled off, the obtained residue was dissolved in 100 ml of DMF,
H-Gly-NH ₂ hydrochloride 1.7 g, NMM 1.7 ml, HOBt 2 g under ice cooling
And 3.4 g of DCC were added. After the mixture was stirred at room temperature for 18 hours, DCUrea was filtered off and DMF was distilled off.

The residue was dissolved in 2-butanol-CH ₂ Cl ₂ (5: 1 v / v), washed successively with saturated aqueous sodium hydrogen carbonate solution, saturated sodium chloride aqueous solution and saturated saline solution, and dried over anhydrous sodium sulfate.

After evaporating the solvent, the residue was crystallized from MeOH-ether and the title compound was collected by filtration.

Yield: 5.0g Melting point: 201-202 ℃ R _f ¹ : 0.26, R _f ² : 0.55 [α] _D : + 2.1 ° (c = 0.5, DMF) (2) Boc-Pro-Arg (Mbs) -Gly -NH ₂ and Z-Arg (Mbs) -Gly- NH 2 20.8g, 80% AcOH 200ml
In the presence of 10% palladium on carbon in a hydrogen stream for 6 hours.

After the palladium carbon was filtered off, the solvent was distilled off. The residue was dried under reduced pressure, dissolved in DMF 200 ml, NMM 4.3 ml,
Boc-Pro-OSu 12.1g was added and it stirred at room temperature for 18 hours.

DMF was distilled off, and the residue was treated with 2-butanol-CH ₂ Cl ₂ (5: 1
v / v), washed successively with saturated aqueous sodium hydrogen carbonate, saturated aqueous sodium chloride and saturated aqueous sodium chloride, and dried over anhydrous sodium sulfate.

After the solvent was distilled off, ether was added to the residue for crystallization, and the title compound was collected by filtration.

Yield: 21.5g Melting point: 120-126 ° C R _f ¹ : 0.31, R _f ² : 0.53 [α] _D : -26.5 ° (c = 1, DMF) (3) Boc-Cys (Acm) -Pro-Arg ( Mbs) -Gly-NH ₂ Boc-Pro-Arg (Mbs) -Gly-NH ₂ 9.8 g, THF 100 ml
After standing in a mixed solvent of 100 ml of 4N HCl-AcOEt for 30 minutes at room temperature, the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in 100 ml of DMF, and added with 3.6 ml of NMM, 5.2 g of Boc-Cys (Acm) -OH, 2.7 g of HOBt and 3.7 g of DCC under ice cooling, and stirred at room temperature for 18 hours.

DCUrea was filtered off, DMF was distilled off, the residue was dissolved in 2-butanol-CH ₂ Cl ₂ (5: 1 v / v), and saturated sodium hydrogen carbonate solution, saturated sodium chloride solution and saturated saline solution were used. After washing sequentially, it was dried over anhydrous sodium sulfate.

After the solvent was distilled off, ether was added to the residue for crystallization, and the title compound was collected by filtration.

Yield: 10.0 g Melting point: 110-116 ° C. R _f ¹ : 0.24, R _f ² : 0.50 [α] _D -58.2 ° (c = 0.5, DMF) (4) Z-pGlu-Cys (Acm) -Pro- Arg (Mbs) -Gly-
NH ₂ Boc-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH ₂ 1.6g
Was left in 10 ml of 4N HCl-AcOEt at room temperature for 30 minutes, and then the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in 20 ml of DMF, and NM under ice cooling.
M (0.22 ml) and Z-pGlu-OSu (0.86 g) were added, and the mixture was stirred at room temperature for 18 hours.

DMF was distilled off, and the residue was treated with 2-butanol-CH ₂ Cl ₂ (5: 1
v / v), washed successively with saturated aqueous sodium hydrogen carbonate, saturated aqueous sodium chloride and saturated aqueous sodium chloride, and dried over anhydrous sodium sulfate.

After the solvent was distilled off, ether was added to the residue for crystallization, and the title compound was collected by filtration.

Yield: 1.4g Melting point: 95-99 ° C R _f ¹ : 0.11, R _f ² : 0.40 [α] _D : -44.7 ° (c = 1.0, DMF) (5) Z-pGlu-Cys (Scm) -Pro- Arg (Mbs) -Gly-
_{NH 2 Z-pGlu-Cys (} Acm) -Pro-Arg (Mbs) -Gly-NH 2
A solution of 1.3 g of CH ₂ Cl ₂ -MeOH (1: 1 v / v) in 80 ml of Cl-Sc under ice cooling.
m (0.22 ml) was added, and the mixture was stirred for 20 minutes.

The solvent was evaporated, the residue was purified by silica gel column with CHCl ₃ -MeOH, crystallized with ether, and the title compound was collected by filtration.

Yield: 540mg Melting point: 185-190 ° C R _f ¹ : 0.19, R _f ² : 0.49 [α] _D : −64.0 ° (c = 1.0, DMF) (6) Z-pGlu-Cys (Scm) -Pro-Arg (Mbs) -Gly-NH 2
Add cysteine hydrochloride 210 mg to 500 mg DMF 10 ml solution,
It was stirred at room temperature for 1 hour.

The solvent was evaporated, the residue was purified by silica gel column with CHCl ₃ -MeOH, crystallized with ether, and the title compound was collected by filtration.

Yield: 400 mg mp: 145~151 ℃ ^{_{(decomposition) R f 2: 0.12 [α}} ] D: -87.0 ° (c = 1.0, DMF) (7) After stirring 150 mg in MSA 2 ml and anisole 0.2 ml at room temperature for 1 hour, ether was added and the supernatant was removed.

The precipitate was dissolved in water, and after treatment with Dowex 1 × 2 (acetate type), water was distilled off.

Dissolve the residue in 0.05% TFA, 12 ml / min (flow rate), 0 to 10% (B) 20 min linear gradient (A) (mobile phase)
After purification by high performance liquid chromatography at
It was treated with 2 (acetate type) and freeze-dried to obtain the title compound.

Yield: 40 mg R _f ³ : 0.11 [α] _D : -160.4 ° (c = 0.5, water) FAB mass spectrum (M + 1): 661 [Example 2] (1) Boc-Cys (MBzl ) The -Pro-Arg (Mbs) -Gly- NH 2 Boc-Pro-Arg (Mbs) -Gly-NH 2 3.7g, 4N HCl-A
After leaving it in 20 ml of cOEt at room temperature for 30 minutes, the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in 50 ml of DMF, and cooled under ice.
0.7 ml of NMM, 2.1 g of Boc-Cys (MBzl) -OH, 0.85 g of HOBt and 1.4 g of DCC were added, and the mixture was stirred at room temperature for 18 hours.

DCUrea was filtered off, DMF was distilled off, the residue was dissolved in CHCl ₃ , washed successively with saturated aqueous sodium hydrogen carbonate solution, diluted hydrochloric acid solution and saturated brine, and dried over anhydrous sodium sulfate.

After the solvent was distilled off, ether was added to the residue for crystallization, and the title compound was collected by filtration.

Yield: 3.2 g Melting point: 104-107 ° C R _f ¹ : 0.44, R _f ² : 0.63 [α] _D : -27.9 ° (c = 0.5, DMF) (2) Z-pGlu-Cys (MBzl) -Pro- Arg (Mbs) -Gly
_{-NH 2 Boc-Cys (MBzl)} -Pro-Arg (Mbs) -Gly-NH 2 2.5
g, 4N HCl-AcOEt 10 ml, NMM 0.4 ml and Z-pGlu-OSu
The title compound was obtained from 1.1 g in the same manner as in Example 1- (4).

Yield: 2.8g Melting point: 108-112 ° C R _f ¹ : 0.22, R _f ² : 0.52 [α] _D -36.0 ° (c = 1.0, DMF) (3) Z-pGlu-Cys (MBzl) -Pro-Arg (Mbs) -Gly-NH 2
140 mg was added to 0.2 ml of anisole and 2 ml of MSA, stirred at room temperature for 1 hour, and then ether was added.

After removing the supernatant and dissolving the precipitate in water, Dowexx1
It was treated with x2 (acetate type) and freeze-dried.

The lyophilized peptide was dissolved in 5 ml of 0.05% TFA, and 33 mg of H-Cys (Scm) -T.hydrochloride prepared in Reference Example 1 was added under ice cooling.

After stirring for 20 minutes, 12 ml / min (flow rate), 10 to 20%
(B) In (A) (mobile phase) of 20-minute linear gradient,
After purification by high performance liquid chromatography, it was treated with Dowex 1 × 2 (acetate type) and freeze-dried to obtain the title compound.

Yield: 32 mg R _f ³ : 0.10 [α] _D : -58.7 ° (c = 0.5, water) [Example 3] (1) cyPent-CO-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH
₂ Boc-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH ₂ 1.5g
Was left in 10 ml of 4N HCl-AcOEt at room temperature for 30 minutes, and then the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in DMF (15 ml), and cooled on ice.
0.32 ml of NMM and cyclopentanecarboxylic anhydride (prepared from 0.48 g of cyclopentanecarboxylic acid and 0.43 g of DCC)
A DMF 2 ml solution was added, and the mixture was stirred at room temperature for 4 hours, and then DMF was distilled off.

The residue was dissolved in 2-butanol-CH ₂ Cl ₂ (5: 1 v / v), washed successively with saturated aqueous sodium hydrogen carbonate solution, saturated sodium chloride aqueous solution and saturated saline solution, and dried over anhydrous sodium sulfate.

After the solvent was distilled off, the residue was purified by silica gel column using CHCl ₃ -MeOH, ether was added for crystallization, and the title compound was collected by filtration.

Yield: 750mg Melting point: 135-138 ° C R _f ¹ : 0.16, R _f ² : 0.45 [α] _D : -54.7 ° (c = 1.0, DMF) (2) cyPent-CO-Cys (Scm) -Pro-Arg (Mbs) -Gly-NH
₂ cyPent-CO-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH
_The title compound was obtained in the same manner as in Example 1- (5) from ₂ 700 mg and Cl-Scm 0.14 ml.

Yield: 640 mg Melting point: 130-133 ° C R _f ¹ : 0.28, R _f ² : 0.55 [α] _D : -65.2 ° (c = 1.0, DMF) (3) cyPent-CO-Cys (Scm) -Pro-Arg (Mbs) -Gly-NH
₂ 600 mg and cysteine hydrochloride 335 mg from Example 1
The title compound was obtained in the same manner as in (6).

Yield: 686mg Melting point: 142-145 ° C R _f ² : 0.18 [α] _D : -77.8 ° (c = 1.0, DMF) (4) 60 mg was treated with MSA-anisole as in Example 1- (7), then 12 ml / min (flow rate), 5 to 25%
(B) In (A) (mobile phase) of 20-minute linear gradient,
After purification by high performance liquid chromatography, it was treated with Dowex 1 × 2 (acetate type) and freeze-dried to obtain the title compound.

^{_{Yield: 29mg R f 3: 0.32 [}} α] D: -167.2 ° (c = 0.5, water) FAB mass spectrum (M + 1): 646 [Example 4] (1) Boc-Pro-Cys (Acm) -Pro-Arg (Mbs) -Gly-
NH ₂ Boc-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH ₂ 1.5g
Was left in 10 ml of 4N HCl-AcOET at room temperature for 30 minutes, and then the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in 20 ml of DMF, and cooled on ice.
Add 0.32 ml of NMM and 0.67 g of Boc-Pro-OSu, and add 1 at room temperature.
After stirring for 8 hours, DMF was distilled off.

The residue was dissolved in 2-butanol-CH ₂ Cl ₂ (5: 1 v / v), washed successively with saturated aqueous sodium hydrogen carbonate solution, saturated sodium chloride aqueous solution and saturated saline solution, and dried over anhydrous sodium sulfate.

After the solvent was distilled off, the residue was purified by silica gel column using CHCl ₃ -MeOH, ether was added for crystallization, and the title compound was collected by filtration.

Yield: 0.6g Melting point: 165-168 ° C R _f ¹ : 0.20, R _f ² : 0.49 [α] _D -83.0 ° (c = 1.0, DMF) (2) Boc-Pro-Cys (Scm) -Pro- Arg (Mbs) -Gly-
NH ₂ Boc-Pro-Cys (Acm) -Pro-Arg (Mbs) -Gly-NH ₂
The title compound was obtained from 450 mg and Cl-Scm of 0.08 ml in the same manner as in Example 1- (5).

Yield: 435 mg Melting point: 205-210 ° C R _f ¹ : 0.33, R _f ² : 0.57 [α] _D : -78.4 ° (c = 1.0, DMF) (3) Boc-Pro-Cys (Scm) -Pro-Arg (Mbs) -Gly-NH ₂
Example 1 from 400 mg and 197 mg of cis-line hydrochloride
The title compound was obtained in the same manner as in (6).

Yield: 416 mg Melting point: 172-180 ° C (decomposition) R _f ² : 0.23 [α] _D -82.1 ° (c = 1.0, DMF) (4) MSA-anisole treatment of 63 mg as in Example 1- (7) followed by 12 ml / min (flow rate), 0-10%
(B) In (A) (mobile phase) of 20-minute linear gradient,
After purification by high performance liquid chromatography, it was treated with Dowex 1 × 2 (acetate type) and freeze-dried to obtain the title compound.

Yield: 22 mg R _f ³ : 0.06 [α] _D : -140.6 ° (c = 0.5, water) FAB mass spectrum (M + 1): 647 [Example 5] (1) Boc-Arg (Mbs) -β-Ala-OBzl H-β-Ala-OBzl.p-toluenesulfonate 3.5 g
Solution of DMF in 50 ml of Et ₃ N 1.4 ml under ice cooling, Boc-Arg (Mbs)
-OH 3.0g, HOBt 1.3g and DCC 1.5g were added.

After stirring at room temperature for 18 hours, DCUrea was filtered off and DMF was distilled off.

The residue was dissolved in AcOEt, washed successively with saturated aqueous sodium hydrogen carbonate solution, diluted hydrochloric acid solution and water, and dried over anhydrous sodium sulfate.

 AcOEt was evaporated to give the title compound as an oil.

Yield: 3.8 g R _f ¹ : 0.55, R _f ² : 0.76 [α] _D : -0.5 ° (c = 1.0, DMF) (2) Boc-Pro-Arg (Mbs) -β-Ala-OBzl Boc-Arg 3.6 g of (Mbs) -β-Ala-OBzl was added to 4N HCl-A.
After standing in 15 ml of cOEt at room temperature for 30 minutes, the solvent was distilled off.

The residue was dried under reduced pressure, dissolved in DMF (50 ml), and NMM (1.0 ml) and Boc-Pro-OSu (2.0 g) were added under ice-cooling.
It was stirred for 8 hours.

DMF was evaporated, the residue was dissolved in AcOEt, washed successively with saturated aqueous sodium hydrogen carbonate solution, diluted hydrochloric acid solution and saturated brine, and dried over anhydrous sodium sulfate.

 AcOEt was evaporated to give the title compound as an oil.

Yield: 3.6 g R _f ¹ : 0.58, R _f ² : 0.75 [α] _D : −28.9 ° (c = 1.0, DMF) (3) Boc-Cys (Acm) -Pro-Arg (Mbs) -β-Ala -O
Bzl Boc-Pro-Arg (Mbs) -β-Ala-OBzl 3.5g, 4N HCl
-AcOEt 15 ml, NMM 0.82 ml and from Boc-Cys (Acm) -OH symmetrical acid anhydride (prepared from Boc-Cys (Acm) -OH 3.2 g and DCC 1.1 g) in Example 3- (1). The title compound was obtained in a similar manner.

Yield: 4.1 g Melting point: 79-83 ° C R _f ¹ : 0.49, R _f ² : 0.74 [α] _D : -27.8 ° (c = 1.0, DMF) (4) Z-pGlu-Cys (Acm) -Pro- Arg (Mbs) -β-Ala-
OBzl Boc-Cys (Acm) -Pro-Arg (Mbs) -β-Ala-OBzl
1.7g, 4N HCl-AcOEt 10ml, NMM 0.3ml and Z-pGlu-O
The title compound was obtained as an oil from Su 0.83 g in the same manner as in Example 1- (4).

Yield: 1.8 g R _f ¹ : 0.43, R _f ² : 0.67 [α] _D : -42.2 ° (c = 1.0, DMF) (5) Z-pGlu-Cys (Scm) -Pro-Arg (Mbs) -β −Ala−
OBzl Z-pGlu-Cys (Acm) -Pro-Arg (Mbs) -β-Ala-
From 1.9 g of OBzl and 0.29 ml of Cl-Scm, Example 1-
The title compound was obtained as an oil in the same manner as in (5).

Yield: 1.2g R _f ¹ : 0.47, R _f ² : 0.73 [α] _D : −64.3 ° (c = 1.0, DMF) (6) Z-pGlu-Cys (Scm) -Pro-Arg (Mbs) -β-Ala-
Example 1 from OBzl 1.0 g and cysteine hydrochloride 0.4 g
The title compound was obtained in the same manner as in (6).

Yield: 980 mg ^{_{mp: 133~137 ℃ R f 2: 0.45}} [α] D: -71.9 ° (c = 1.0, DMF) (7) 70 mg was treated with MSA-anisole as in Example 1- (7), then 12 ml / min (flow rate), 0 to 10%
(B) In (A) (mobile phase) of 20-minute linear gradient,
After purification by high performance liquid chromatography, it was treated with Dowex 1 × 2 (acetate type) and freeze-dried to obtain the title compound.

^{_{Yield: 27mg R f 3: 0.14 [}} α] D: -154.0 ° (c = 0.5, water) FAB mass spectrum (M + 1): 676 [Example 6] H-Pro-Cys- Pro-Arg-Gly-NH 2 -Acetate (1) Z-Pro-Cys (MBzl) -Pro-Arg (Mbs) -Gly-
NH ₂ Boc-Cys (MBzl) -Pro-Arg (Mbs) -Gly-NH ₂ 2.4
g, 4N HCl-AcOEt 10 ml, NMM 0.6 ml and Z-Pro-OSu
The title compound was obtained from 1.3 g in the same manner as in Example 1- (4).

Yield: 2.3 g Melting point: 101-104 ° C. R _f ¹ : 0.41, R _f ² : 0.61 [α] _D -56.4 ° (c = 1.0, DMF) (2) H-Pro-Cys-Pro-Arg-Gly -NH ₂ · acetic acid salt Z-Pro-Cys (MBzl) -Pro-Arg (Mbs) -Gly-NH 2
After treating 150 mg with MSA-anisole as in Example 1- (7), 12 ml / min (flow rate), 0-10%
(B) In (A) (mobile phase) of 20-minute linear gradient,
After purification by high performance liquid chromatography, it was treated with Dowex 1 × 2 (acetate type) and freeze-dried to obtain the title compound.

Yield: 64 mg R _f ³ (containing 0.1% ethanediol): 0.12 [α] _D : -92.7 ° (c = 0.5, water) FAB mass spectrum (M + 1): 528 [Example 7] H-Pro-Cys-Pro-Arg-Gly-NH ₂ · acetic acid salt 30 mg in water 2
It was dissolved in ml and adjusted to pH 7 with dilute aqueous ammonia. The mixture was stirred at room temperature for 7 days, acidified with acetic acid, and lyophilized.

Yield: 28 mg R _f ³ : 0.02 [α] _D : -142.9 ° (c = 0.5, water) FAB mass spectrum (M + 1): 1054 Next, a pharmacological test example showing the effectiveness of the peptide derivative of the present invention. Show.

[Pharmacological test example] The effect on memory fixation was examined by using male Wistar rats, [Burbach] et al. [Science (Scienc
e), 221 , 1310-1312 (1983)]. The experimental apparatus consists of a light room and a dark room, and the floor is made of a stainless steel grid. Rats placed in the lightroom are free to move to the darkroom. Using this device, rats undergo a single electrical shock when they enter the darkroom. The retention of the passive avoidance behavior to the electric shock was determined by the time (reaction latency) until a rat placed again in the light room after a certain period of time entered the dark room.

Examination of Experimental Improvement Effect of Cycloheximide on Experimental Retrograde Amnesia After the peptide derivative of the present invention or physiological saline was subcutaneously administered, an electric shock (0.5 mA) was caused 1 hour later, and immediately after that, cycloheximide 2.7 to 3.0 mg / kg or physiological saline was subcutaneously administered, and a memory retention test was performed 48 hours later.
Rats to which only saline was administered generally exhibited a response latency of about 300 seconds, and rats of the control group to which cycloheximide alone was administered exhibited a response latency of about 50 seconds, and developed retrograde amnesia.

The average value of the reaction latency of the peptide derivative administration group of the present invention was compared with that of the control group. The number of rats used in the tests in each group is 6-8. The maximum measurement time was 600 seconds.

Table 1 shows the doses and effects of the peptide derivatives obtained in each example (the ratio of the reaction latency of each example to the reaction latency of the control group is shown in%).

From the above test results, it is clear that the peptide derivative of the present invention has an excellent effect of improving retrograde amnesia.

Next, formulation examples of drugs containing the peptide derivative of the present invention are shown.

[Formulation Example 1] (Injection) In 100 ml of distilled water for injection, 0.1 mg of the peptide derivative obtained in Example 1 and 0.9 g of sodium chloride were added, and p
An aqueous solution in which H was adjusted to 6.0 to 8.0 with sodium hydroxide was prepared. This was filtered through bacteria, filled in a 1 ml ampoule, sealed, and sterilized by heating to produce an injection.

[Formulation Example 2] (Freeze-dried preparation) 5 ml of the peptide derivative obtained in Example 1 and 5 g of D-mannite were added to 100 ml of distilled water for injection, and the pH was adjusted to 6.0 to 8.0 with a phosphate buffer. A conditioned aqueous solution was prepared.
This was subjected to bacterial filtration, dispensed in a vial (1 ml), and lyophilized to produce a lyophilized injection.

[Formulation Example 3] (Nasal drops) 10 mg of the peptide derivative obtained in Example 1 was added to 100 ml of physiological saline, and the pH was adjusted to 3.0 to 6.0 with a citrate buffer.
To 50 μg in a single dose of 0.5 ml to prepare a nasal drop.

[Formulation Example 4] (suppository) Hard fat (triglyceride of saturated fatty acid) 9
After adding 0.5 g of egg yolk lecithin to 8.5 g and melting at 40 to 45 ° C., 5 mg of the peptide derivative obtained in Example 1 was added to PEG400.
After adding 1g of the liquid dissolved in this and stirring and dispersing,
1 g thereof was poured into a suppository mold, and after solidification, it was separated from the mold to produce a suppository.

[Effects of the Invention] The peptide derivative of the present invention is a novel compound, has an excellent nootropic action, and is useful as a medicine.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical display location C07K 11/00 (72) Inventor Yoshiharu Nakajima 4-6-7 Shimochiai, Shinjuku-ku, Tokyo Fujirebio shares In-house (72) Inventor Mitsuo Masaki 5-11-6 Masago, Chiba-shi, Chiba Prefecture (72) Masaki Uehara 1340-2 Hiranuma, Yoshikawa-cho, Kitakatsushika-gun, Saitama Kenji Hirate Masuda Nitta, Kasukabe-shi, Saitama Prefecture 407-9

Claims (2)

[Claims] 1. The following general formula (I): [Wherein A is cyclopentylcarbonyl, Pro or pGlu
And B is Gly or β-Ala, W is a hydrogen atom or the following general formula (II): (In the formula, Y ¹ is H or CO-T, and Y ² is OH or T (where T is the following general formula (III): (In the formula, R ¹ is a group selected from the group consisting of an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to 10 carbon atoms, and an alkylthio group having 1 to 6 carbon atoms) Or the following general formula (IV): (Wherein R ² is a group selected from the group consisting of a hydrogen atom, an alkylcarbonyl group having 2 to 7 carbon atoms, and an arylcarbonyl group having 7 to 10 carbon atoms), )) Is a group represented by] or a peptide derivative represented by: or a general formula (V): [Wherein A and B are the same as defined above], or a derivative in their functional group, or a pharmacologically acceptable salt thereof. 2. The following general formula (I): [Wherein A is cyclopentylcarbonyl, Pro or pGlu
And B is Gly or β-Ala, W is a hydrogen atom or the following general formula (II): (In the formula, Y ¹ is H or CO-T, and Y ² is OH or T (where T is the following general formula (III): (In the formula, R ¹ is a group selected from the group consisting of an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to 10 carbon atoms, and an alkylthio group having 1 to 6 carbon atoms) Or the following general formula (IV): (In the formula, R ² is a group selected from the group consisting of a hydrogen atom, an alkylcarbonyl group having 2 to 7 carbon atoms and an arylcarbonyl group having 7 to 10 carbon atoms), )) Is a group represented by] or a peptide derivative represented by: or a general formula (V): [Wherein A and B are the same as above], or an effective amount of a peptide derivative represented by a functional group, or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable salt. An anti-dementia agent comprising the obtained carrier or diluent.