JPH02273694A - Peptide derivative and antidemential agent containing the same - Google Patents

Abstract

NEW MATERIAL:The compound of formula I {A is cyclopentylcarbonyl; Pro is pGlu; B is GLy or B-Ala; W is H or group of formula II [Y<1> is H or CO-T; Y<2> is OH or T (T is group of formula III (R1 is 2 to 7C alkylcarbonyl, 7 to 10C arylcarbonyl or 1 to 6C alkylthio) or formula IV (R<2> is H, 2 to 7C alkylcarbonyl or 7 to 10C arylcarbonyl))]}. EXAMPLE:The compound of formula V. USE:It has nootropic action and is useful as a drug such as antidemential agent. PREPARATION:Condensation reaction is carried out by azide process, etc., in a solvent (e.g. chloroform) at -30-+50 deg.C while protecting a group not partici pating to in the reaction or activating a group participating in the reaction.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to peptides and peptide derivatives that have nootropic effects and are therefore useful as medicines, particularly as anti-dementia agents.

[Prior Art] It has been known for a long time that Pap pressin has a nootropic effect, but recently peptides that can be considered to be fragments of Pap pressin, such as pGfu-Asn-Cys-Pro-＾rg-Gly-
Ni12H-Cys-0)1.

Also, ■-^5n-Cys-Pro-^rg-OH1I-Cy
It has been reported that the peptide represented by s-OH also has a nootropic effect similar to pap pressin [Science
ce) 221.1310-1312 (1983)
] [Brain Research (Brain Re5ear
ch) 371.17 (198B)].

[Problems to be Solved by the Invention] The purpose of the present invention is to provide novel peptides and peptide derivatives that have even better nootropic effects than such Pappressin and Pappressin fragment peptides. be.

[Means for Solving the Problem] The present invention provides the following formula (■): A -Cys-Pro-8rg-B
(I) [wherein A is cyclopentylcarbonyl, Pr.

or pGlu, B is Gly or β-Ala, W is a hydrogen atom or the following formula (■): Y'-[:ys-Y" (II) (
In the formula, Yl is H or Co-T, Y2 is OH or T
(wherein, T is the following formula (■): (wherein, R1 is an alkylcarbonyl group having 2 to 7 carbon atoms, an arylcarbonyl group having 7 to 10 carbon atoms, and an arylcarbonyl group having 1 to 6 carbon atoms) (a group selected from the group consisting of alkylthio groups), or the following general formula (■): (wherein R2 is a hydrogen atom, an alkylcarbonyl group having 2 to 7 carbon atoms, or a group having 7 to 10 carbon atoms) A group selected from the group consisting of arylcarbonyl groups), a group represented by)) a peptide derivative represented by the following, or a peptide derivative represented by the following general formula (V): (A-Cys -Pro-＾rg-B)t
(V) The present invention relates to a peptide derivative represented by the formula [wherein A and B are the same as above], or a functional group derivative 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 functional group derivative thereof, or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable carrier or The present invention relates to an anti-dementia agent containing a diluent.

The functional group derivative 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 1,000-alkyl- or di-alkyl-substituted amides having alkyl groups of 1 to 6 carbon atoms; and c) esters derived from alcohols having 1 to 18 carbon atoms, preferably aliphatic alcohols having 1 to 6 carbon atoms.

Pharmaceutically acceptable salts of the above-mentioned peptide derivatives or their functional group derivatives include acid addition salts and basic salts. Such acid addition salts include salts of inorganic acids (e.g., hydrochloric acid, sulfuric acid, phosphoric acid) or organic acids (e.g., acetic acid, propionic acid, citric acid, tartaric acid, malic acid, oxalic acid, methanesulfonic acid), etc. Can be mentioned. Further, examples of the basic salt include sodium salt, potassium salt, triethylamine salt, and the like.

In this specification, amino acids, peptides, protecting groups, solvents, etc. are referred to by abbreviations commonly used in the art, or IU
Abbreviations adopted by the PAC-IUB nomenclature committee are used. For example, the following abbreviations are used: In addition, amino acids shall mean L-type amino acids.

β-Ala: β-alanine Arg: Arginine Cys Nijistine Guy Nigericin pGlu: Pyroglutamic acid Pro Nibroline Boc: t-Butoxycarbonyl Z: Benzyloxy carbonyl Mbs: P-methoxybenzenesulfonyl MBzl:
P-methoxybenzyl Acm: Acetamidomethyl Sctm: Carbomethoxysulfenyl 0Bzl: Benzyl ester O5u: N-hydroxysuccinimide ester DCC
: N,N'-dicyclohexylcarbodiimide DCU
rea: N,N'-dicyclohexylurea HOB
t: 1-Hydroxybenzotriazole εt3N Nitriethylamine NMM: N-methylmorpholine TFA: Trifluoroacetic acid MSA: Methanesulfonic acid Ac0Et: Ethyl acetate^cOH: Acetic acid T) IF: Tetrahydrofuran DMF: N,N-dimethylformamide MeOH: Methanol The peptide derivatives of the present invention can be prepared by methods commonly used in peptide chemistry, such as 5chr2ideran.
d Li1bke, “The Peptide”
ides) J Vol.-, Academic Press,
New York, tl, S, A, (1965), Nobuo Izumiya et al., “Basics and Experiments of Peptide Synthesis”, Maruzen @1 (1
985) (liquid phase method and solid phase method).

Condensation methods for forming peptide bonds include azide method, acid chloride method, acid anhydride method, 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 carrying out the condensation reaction, carboxyl groups, amino groups, etc. that do not participate in the reaction may be protected, or carboxyl groups, amino groups that participate in the reaction may be activated by means known per se.

Examples of the carboxyl protecting group include esters such as methyl, ethyl, benzyl, p-nitrobenzyl, t-butyl, and cyclohexyl.

Examples of protecting groups for amine groups include benzyloxycarbonyl group, t-butoxycarbonyl group, isobornyloxycarbonyl group, and 9-fluorenylmethoxycarbonyl group.

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

Examples of protecting groups for mercapto groups include trityl group,
Examples include acetamidomethyl group, benzyl group, p-methoxybenzyl group, and 3-nitro-2-pyridinesulfenyl group.

Examples of activated carboxyl groups include:
Corresponding acid anhydrides, azides, active esters [alcohols (e.g., pentachlorophenol, 2,4-dinitrophenol, cyanomethyl alcohol, p-nitrophenol, N-hydroxy-5-norbornene-2,3-dicarboximide) , N-hydroxysuccinimide, N-
esters with hydroxyphthalimide, 1-hydroxybenzotriazole), and the like. Examples of activated amino groups include the corresponding phosphoric acid amides.

The reaction is usually carried out in a solvent, such as chloroform, dichloromethane, ethyl acetate, N,N-dimethylformamide, dimethyl sulfoxide, pyridine, dioxane, tetrahydrofuran, water, methanol, etc.
Alternatively, it can be carried out in a mixture of these.

The reaction temperature can be within the commonly used range of about -30°C to about 50°C.

The protecting group removal reaction for the peptide of the present invention varies depending on the type of protecting group used, but it is necessary that the protecting group be removed without affecting the peptide bond.

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

After the reaction, the peptides and peptide derivatives of the present invention produced in this way can be obtained by known peptide separation methods such as extraction, distribution, reprecipitation, recrystallization, column chromatography, etc. can.

Furthermore, the peptides and peptide derivatives of the present invention can be made into functional group derivatives or pharmacologically acceptable salts thereof, as described above, by methods known per se.

The peptide derivatives of the present invention exhibit strong nootropic effects in passive avoidance tests in rats.

Useful target diseases for the peptide derivatives of the present invention include:
For example, senile dementia (Alzheimer's type dementia), cerebrovascular dementia, and dementia based on Alzheimer's disease, Pick's disease, Huntington's chorea, Kreuffelt-Jakob disease, Parkinson's disease, myelinopathy of the cerebellum, etc. 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 have been no cases of death even at doses far exceeding the therapeutically effective dose.

The peptide derivatives of the invention can be administered as free forms or as derivatives at their functional groups.

The appropriate dosage for administration is generally in the range of 0.1 ng to 1 mg/day in terms of the amount of the free form, whether it is the free form or its salt. In particular, parenteral and nasal administration
It is preferably 0.1 ng to 100 μg/day, and in oral administration or rectal administration, it is preferable to administer 10 to 100 times as much as parenteral administration.

The peptide derivative of the present invention is mainly administered parenterally (e.g., intravenous or subcutaneous injection, intracerebroventricular or intrathecal administration, nasal administration, rectal administration), but in some cases, it may be administered orally. Good too.

Examples of dosage forms include injections, suppositories, powders, nasal sprays,
Examples include pills and tablets. The peptide derivative of the present invention can be stored as a solution in physiological saline, but it can also be prepared into a lyophilized ampoule by adding mannitol or sorbitol, and then dissolved at the time of use.

Examples are shown below.

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

R, I, chloroform-methanol-acetic acid-water (80:
20:2.5:5) Lower layer R, 2, chloroform-methanol-water (70:30:
5) R,3:n-butanol-acetic acid-water (2:1:1) Also, special preparation by high performance liquid chromatography is as follows: Column: μBondapak C,! , 9 x 1
Performed using 5cm mobile phase: A) 0.05% TFA, B) Acetonitrile.

[Reference Example 1] H-Gys (Scts) -T hydrochloride (however, T
is the formula (I) in which l(+ is a benzoyl group).
II) (1) Bocl;y
s (AcIl) Boc-Cys (Acm) -OH
1, Og%S-benzoylthiamine 1.3g and 4
-dimethylaminopyridine 20mg (:82C125
Dtl under water cooling to 0 m 11 solution: (: 0.78 g C
A warm solution of 112CI2 5 m It was added dropwise.

After stirring for 30 minutes under water cooling and further stirring at room temperature for 1 hour, DC
Urea was filtered off and washed with saturated sodium bicarbonate water and water.

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

Yield: 1.8g Melting point = 71-75°C Re': 0. 74 Rt2: 0.82 [a
] o Knee 39.2° (cwo, 5. DMF) (
2) Boc-Gys (SC!I) -TBoc-
Cys(Acm)-T 600 mg MeOH-C
H2Cl2 (1: lv/v) 6ml! Cl in solution
-5 cm 0.14 m 11 was added and stirred at room temperature for 20 minutes.

After distilling off the solvent, the residue was purified using a silica gel column using (:HGI3-MeO)1 to obtain the title compound as an oil.

Yield: 580mg Rt': 0.82 Rr2: o, sa[a]
o Knee 44.0″″ (c-0,5,DMF) (3
) It-(:ys (Scm) -T Hydrochloride B
oc-Cys (Scm) -T 470m g 4
After standing in 2 ml of N HGI-AcOEt at room temperature for 30 minutes, the solvent was distilled off. The residue was converted into CHGI3-Me
Purification was performed on a silica gel column using ol() to obtain the title compound as an oil.

Yield: 250mg Ry': 0.38 Rr": o, 5
4 [alo: +38.4° (clIo, 5.
DMF) [Example 1] H-Cys-OH ■ pGIu-(:ys-Pro-^rg-Gly-Nu,
・Acetate (1) Z-Arg(Mbs)-Gly-N
After stirring and dissolving 1°g of H2Z-Arg(Mbs)-0H dicyclohexylamine salt in 100ml of AcoEt + 7°ml of 5% citric acid water, the AcoEt layer was washed with water and dried over anhydrous sodium sulfate.

The solvent was distilled off, and the resulting residue was dissolved in DMF 100 m
J! H-Gly-NI2 hydrochloride 1.
7g, NMM 1.7ml! , 2 g HOBt and 3.4 g DCG were added. Mixture at room temperature
After stirring for an hour, DCUrea was filtered off and DMF was distilled off.

The residue was dissolved in 2-butanol-CI2C12 (5:1 v
/v), washed successively with saturated aqueous sodium bicarbonate, saturated aqueous dilute hydrochloric acid, and saturated brine, and then 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: 1 it: 5. Og Melting point = 201-202°C Rr': o, 26. Rr2+ 0.55 [a]
, : +2. 1” (c-0,5,DMF)(
2) Boc-Pro-Arg(Mbs)-Gly-N
)12Z-Arg(Mbs)-Gly-NH,20,8
g was stirred in 80% AcOAcOH20O in the presence of 10% palladium on carbon for 6 hours under a stream of hydrogen.

After filtering off the palladium on carbon, the solvent was distilled off. After drying the residue under reduced pressure, it was dissolved in DMF 200 m It, NMM 4.3 ml, Boc-Pro-O5
12.1 g of u was added, and the mixture was stirred at room temperature for 18 hours.

DMF was distilled off, and the residue was dissolved in 2-butanol-CH2 (:
12 (5:1 v/v), washed successively with saturated sodium bicarbonate water, salt-saturated dilute hydrochloric acid water, and saturated salt water, and then dried over anhydrous sodium sulfate.

After evaporating the solvent, ether was added to the residue to crystallize it, and the title compound was collected by filtration.

Yield: 21.5g Melting point: 120-126°C Rf': o, 31, Rr2: o, 53[
(!lo knee 26.5″″ (c=1, DMF)
(3) Boc-(:ys(Acm)-Pro-Arg
(Mbs) -Gly-Nl(2Boc-Pro-Ar
g(Mbs)-Gly-NH, 9.8g, T)IF
100 mft and 4 N HCl-Ac0Et 100
m j! After leaving it for 30 minutes at room temperature in a mixed solvent with
The solvent was distilled off.

After drying the residue under reduced pressure, it was dissolved in 100 m 11 of DMF and diluted with NMM 3.6 m B, % Boc- under water cooling.
Cys(Act)-0H5, 2g, HOBt 2.
7 g and 3.7 g of DCC were added, and the mixture was stirred at room temperature for 18 hours.

DCUrea was filtered off, DMF was distilled off, and the residue was
Butanol-(:I2(:12 (5:1 v/v)
The solution was washed successively with saturated aqueous sodium bicarbonate, dilute hydrochloric acid saturated with sodium chloride, and saturated brine, and then dried over anhydrous sodium sulfate.

After evaporating the solvent, ether was added to the residue to crystallize it, and the title compound was collected by filtration.

Yield: 10. Og Melting point: 110-116°C Re': 0. 24. R,2: 0.
50 [α]. Knee 58. 2° (c-0,5,D
MF) (4) Z-pGIu-(:ys(Ac11)
-Pro-Arg(Mbs)-Gly-NH2Boc-
(:ys(Ac+*)-Pro-Arg(Mbs)-G
ly-NI21.6 g to 4NIIC1-^cOεtl
After standing in omJZ for 30 minutes at room temperature, the solvent was distilled off.

The residue was dissolved in vacuum-dried iJDMF 20 m It and NMM O, 22 m J! was added under ice cooling. , Z-pGIu
-O5u O, 86 g was added and stirred at room temperature for 18 hours.

DMF was distilled off, and the residue was dissolved in 2-butanol-CH2 (:
12 (5:1 v/v), washed successively with saturated aqueous sodium bicarbonate, saturated dilute hydrochloric acid and saturated brine, and dried over anhydrous sodium sulfate.

After evaporating the solvent, ether was added to the residue to crystallize it, and the title compound was collected by filtration.

Yield: 1.4g Melting point = 95-99°C R,': 0.11. Rr2: 0.40 [α]. :
-44,7@(c-1,0,DMF) (5) Z-p
Glu-Cys(Scm)-Pro-Arg(Mbs)
-Gly-Nt-12Z-pGlu-Cys(Ac+s
)-Pro-Arg(Mbs)-Gly-NH,1,3
g of CH2CIz-MeO)1 (1:1 v/v)
80 m iL warm solution under ice cooling C1-5 cm O, 22
ml! was added and stirred for 20 minutes.

The solvent was distilled off, and the residue was purified on a silica gel column using C) lc13-MeOH, crystallized from ether, and the title compound was collected by filtration.

Yield + 540mg Melting point = 185~! 90°C R,': 0.19. R,': 0.49 [α] o
: -64,0" (c-1,,o, DMF) (6)
H-(:yS-OH Z-pGIu-11:ys-Pro-Arg(Mbs)
-Gly-NH2・hydrochloride 7, -pGlul:ys(S
cm)-Pro-Arg(Mbs)-Gly-NH25
210 mg of cysteine hydrochloride was added to a solution of 00 rng in 10 mffi of DMF, and the mixture was stirred at room temperature for 1 hour.

The solvent was distilled off, and the residue was purified on a silica gel column using CuCl2-MeOH, crystallized from ether, and the title compound was collected by filtration.

Yield: 400 mg Melting point = 145-151°C (decomposed) R,': 0.12 [α]o Knee 87. Oo (c-1,0,DMP)
)1-fl:ys-OH Z-pGlu-Cys-Pro-Arg(Mbs)-G
ly-N)lx ・150 mg of hydrochloride was added to 2 mI of MSA
After stirring for 1 hour at room temperature in L and 0.2 m It of anisole, ether was added and the supernatant was removed.

The precipitate was dissolved in water, and after treatment with DowexlX2 (acetate type), water was distilled off.

The residue was dissolved in 0.05% TFA and purified by high performance liquid chromatography in (A) (mobile phase) with a 20 min linear gradient from 0 to 10% (B) at i 2 ml 1/min (flow rate). , treated with DowexlX2 (acetate form) and lyophilized to give the title compound.

Yield: 40 mg Rr3: 0.11 [α]. knee 160.4° (c-0,5, water) FAB mass spectrum (M+1) +661 [Example 2 H-Cys-T pGIu-(:ys-Pro-Arg-Gly-NH2
-Acetate (1) Boc-Cys(MBzl)-Pro
-Arg(Mbs)-Gly-NH.

Boc-Pro-Arg(Mbs)-Gly-NH23
, 7 g, 20 m fl of 4N HCl-8 cOEt,
After being left in the solution for 30 minutes at room temperature, the solvent was distilled off.

After drying the residue under reduced pressure, it was dissolved in DMF 50 mJ2 and cooled with NMM 0.7 m 11 , Boc-Cys.
(MBzl)-0H2, 1g, HOBt O, 85g
and 1.4 g of DCC were added, and the mixture was stirred at room temperature for 18 hours.

D(: Urea was filtered off, DMF was distilled off, and the residue was
The solution was dissolved in 11 fl;

After evaporating the solvent, ether was added to the residue to crystallize it, and the title compound was collected by filtration.

Yield: 3.2g Melting point = 104-107°C Rr': o, 44 Rf': 0.63 [α]
. , -27,9° (c-0,5,DMF) (2) Z
-pGlu-tl:yS(MBzl)-Pro-Arg
(Mbs) -Guy-NH2Boc-Cys (MB
zl)-Pro-8rg(Mbs)-Gly-NH22
,5g, 4NHCl-Ac0Et 10mIL,
NMIA O, 4 ml and Z-pGlu-O5u 1
．． 1 g, as in Example 1-(4),
The title compound was obtained.

Yield: 2.8g Melting point: 108-112°C Rr': o, 22. Re': 0. 52
[α]. : -36,0' (cd, o, DMF) (
3) H-11:ys-T ■ pGlu-Cys-Pro-Arg-Gly-Nil,
-acetate Z-pGlu- to ys(MBzl)-Pro-
Arg(Mbs)-Gly-NH, 140 mg, was added to 0.2 m fl of anisole and MSAZmIL, and after stirring at room temperature for 1 hour, ether was added.

After removing the supernatant and dissolving the precipitate in water, Dowex
lx2 (acetate type) treatment and lyophilization.

The lyophilized peptide was dissolved in 0.05% TFA5mJ2, and H-Cys (Scm) -T produced in Reference Example 1 was dissolved.
- 33 mg of hydrochloride was added under water cooling.

After stirring for 20 min, Dow
treatment with exlX2 (acetate form) and lyophilization afforded the title compound.

Yield: 32 mg Rf3: 0.10 [α]o: 5B, 7° (c-0,5, water) [Example 3] H-Cys-0)1 cyPent-fl:0-Cys-Pro-Arg- G
ly-N)+2・acetate (cyPenL dichloropentyl group) cyPent-GO-C:ys(Ac11)-Pro-
Arg(Mbs)-G13/-NH2Boc-(:ys
(Acm)-Pro-Arg(Mbs)-Gly-N
l21.5 g to 4N II[:1-AcOEt1
After standing for 30 minutes at room temperature in 0 ml, the solvent was distilled off.

After drying the residue under reduced pressure, it was dissolved in DMF 15mJ2, and under water cooling, NMM 0.32mJ2 and cyclopentanecarboxylic anhydride (cyclopentanecarboxylic acid 0.48g
A solution of 2mu of DMF (prepared from 43 g of DCG O and 43 g of DCG O) was added thereto, and after stirring at room temperature for 4 hours, DMF was distilled off.

The residue was dissolved in 2-butanol-(:l12C12(5:1
(v/v), washed successively with saturated aqueous sodium bicarbonate, saturated aqueous dilute hydrochloric acid, and saturated brine, and then dried over anhydrous sodium sulfate.

After evaporating the solvent, the residue was subjected to a special silica gel column using CHCI 3-MeOll, and ether was added to crystallize it, and the title compound was collected by filtration.

Yield: 50 mg Melting point: 135-138°C R,': 0.16. R,': 0.45[α]
o Knee 54.7° (C ~ 1.0.DMF) cyPe
nt-fl:0-Cys(Scm)-Pro-Arg
(Mbs) -G Iy-N[1゜cyPenL-GO
-Cys(8cm)-Pro-Arg(Mbs)-Gl
y-882700mg and Cl-5cm 0.14 m
The title compound was obtained in the same manner as in Example 1-(5), yield: 640 mg, melting point = 130-133°C, R,': 0. 28. Rf2:0. 55[α
]. Knee 65.2° (c-1,0, DMF) (3
) To(:ys-01-1 cyPent-GO-Cys-Pro-Arg(Mbs
)-Gly-Nl2-hydrochloride cyl'en L-GO-
Cys(Scm)-Pro-Arg(Mbs)-Gl
y-N82600mg and cysteine hydrochloride 3:15m
The title compound was obtained in the same manner as in Example 1-(6).

Yield: 686 mg Melting point = 142-145°C R,2:0.18 [α]o Knee 77.8° (c-1,0,DMF)
(4) It-Cys-OH cyPent-GO-Cys-Pro-^rg-Gly
-NH2 Acetate II-Cys-OH cyPent-(:O-Cys-Pro-Arg(Mb
s)-G! Example 1 - 60 mg of y-Nll□ hydrochloride
After MS 8-anisole treatment as in (7), 12 mfl 1 min (flow rate), 5 to 25% (B)2
After high performance liquid chromatography using (A) (mobile phase) of a 0 minute linear gradient, DowexlX2 (
acetate form) and lyophilization to give the title compound.

Yield: 29 mg R, 3:0.32 [α]. Knee 167.2° (c-0,5, water) FAB mass spectrum (M+1): 646 [Example 4] (1) Boc-Pro-Gys (Acm)-Pro
-8rg(Mbs)-Gly-NH□Boc-Cys
(8cm)-Pro-Arg(Mbs)-Gly-Nt
12 1.5 g to 4 N tLcl-AcOEt
After standing in 10 m fl for 30 minutes at room temperature, the solvent was evaporated.

After drying the residue under reduced pressure, it was dissolved in DMF20mj2 and mixed with NMM 0.32m fl and Boc-Pro-
After adding 67 g of O5uO and stirring at room temperature for 18 hours, DMF was distilled off.

The residue was dissolved in 2-butanol-CH□Cl2 (5:1 v
/v), washed successively with saturated aqueous sodium bicarbonate, saturated aqueous dilute hydrochloric acid, and saturated brine, and then dried over anhydrous sodium sulfate.

After the solvent was distilled off, the residue was purified through a special silica gel column using CHC13-MeOH, and ether was added to crystallize it, and the title compound was collected by filtration.

Yield: 0.6g Melting point: 165-168°C R11,0,20,R,2:0.49 [a]o: 83.0° (c=1.o, DMF)
(2) Boc-Pro-Cys(Scm)-Pro-
Arg(Mbs)-Gly-NH.

Boc-Pro-11:ys (8cm)-Pro-A
rg(Mbs)-Gly-NH2450mg and Cl-
From 5crtp O, 08m fl, Example 1-(5
) The title compound was obtained in the same manner as in ).

Yield: 435 mg Melting point: 205-210°C R, I: 0. 33. R, 2:0. 57[α
] o Knee 78. 4° (c-1,0,DMF) [
]]oc-Pro-C:ysScffl)-Pro-A
The title compound was obtained from 400 mg of rg(Mbs)-Gly-NH□ and 197 mg of cysteine hydrochloride in the same manner as in Example 1-(6).

Yield: 416 mg Melting point = 172-180°C (decomposed) Ry': 0.23 [α. Knee 82. 1° (c-1,0,DMF
)(4) H-Cys-OH H-Pro-C:ys-Pro-Arg-Gly-NH
2.Acetate)1-Cys-0)1 ~ Boc-Pro-Gys-Pro-Arg(Mbs)-
Example 1-(7
After MSA-anisole treatment as in ), high-performance liquid chromatography loading was performed with (A) (mobile phase) at 12 mfl/min (flow rate) and a 20 min linear gradient of 0 to 610% (B). Thereafter, it was treated with DowexlX2 (acetate type) and lyophilized to obtain the title compound.

Several groups: 22 mg R, 3: 0.06 [α co. Knee 140, 6° (c-
0,5, water) FAB mass spectrum (M-11): 64
7 [Example 5 (1) Boc-Arg(Mbs)-β-Ala-O
Ei was added to a solution of 3.5 g of Bz11t β-Ala-OBzl-P-toluenesulfonate in 50 mJZ of DMF under water cooling.
3N 1.4ml1. , Boc-Arg(Mbs)-O
ff 3.0 g, tlOBt 1.3 g and DCC
1.5 g was added.

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

The residue was dissolved in Ac0Et, washed successively with saturated aqueous sodium bicarbonate, diluted hydrochloric acid, and water, and then dried over anhydrous sodium sulfate.

8 cOEt was distilled off to give the title compound as an oil.

Yield: 3.8g Rr': 0. 55. Ry': 0. 76 [
α】o: 0.5° (c-1,0,DMF) (2)
Boc-Pro-Arg(Mbs)-β-Ala-OB
zlBoc-Arg(Mbs)-β-Ala-OBzl
3.6 g of 4NHCI-AcOEt 15 m
After standing for 30 minutes at room temperature in 100 ml of water, the solvent was distilled off.

After drying the residue under reduced pressure, it was dissolved in DMF50rnIL and diluted with NMM 1. under water cooling. Om It and Boc-Pro
-O5u 2. Og was added and stirred at room temperature for 18 hours.

DMF was distilled off, and the residue was dissolved in AcOEt, washed successively with saturated aqueous sodium bicarbonate, diluted hydrochloric acid, and saturated brine, and then dried over anhydrous sodium sulfate.

Ac0Et was distilled off to obtain the title compound as an oil.

Collection jl: 3.6g Fly': 0.5B, Re2: 0.75[α
]. Knee 28.9° (c-1,0,DMF) (3)
Boc-Cys(Acm)-Pro-Arg(Mbs)
-β-Ala-OBzlBoc-Pro-Arg (M
bs)-β-^1a-OBzl 3.5 g, 4NHC
I-^cOEt 15 ml, NMM 0.82
ml and Boc-Cys (3cm)-OH symmetric acid anhydride (Boc-Cys (Acm)-0H3,2g and D
C (prepared from: 1.1 g), Example 3(1)
The title compound was obtained in the same manner as in .

Yield: 4.1g Melting point near 9-83°C Rf': 0. 49. RF2: 0. 7
4[αlo knee 27.8° (c-1,0,DMF
)Z-pGIu-Cys (Acm)-Pro-Ar
g (Mbs)-β-Ala-OBzlBoc-11:
ys (Acm) -Pro-Arg (Mbs)-β
-Ala-OBzl 1.7g, 4N HCl-^
cOEtlOmj! , NMM 0.3 m Il and Z
Example 1-(4
) The title compound was obtained as an oil.

Collection: t, sg R,': 0.43. R,2: 0.67[α
] B: -42,2" (c-1,0,D
MF) Z-pGlu-Cys (Sew)-Pro-A
rg(Mbs)-β-Ala-OBzlZ-pGlu-
Cys (3cm)-Pro-Arg(Mbs)-β-
Ala-OBz11.9 g and Cl-5cm 0.2
9 m It, the title compound was obtained as an oil in the same manner as in Example 1-(5).

Yield 1i: 1.2g Rf': 0.47. Rf2: 0.73 [α]
o knee 64.3° (c-1,0,DMF)H-(
:ys-OH Z-pGIu-Cys-Pro-Arg(Mbs)-β
-Ala-OBzl Hydrochloride Z-pGlu-Cys
(Scm)-Pro-Arg(Mbs)-β-Ala
Example from -OBzlt, o g and cysteine hydrochloride 0.4 g! - The title compound was obtained in the same manner as in (6).

Yield: 980 mg Melting point: 133-137°C R, 0.45
70 mg of salt was added to M as in Example 1-(7).
After SA-anisole treatment, 12 m 11 minutes (flow 1 t)
, 10% (B) from O to 20 min linear gradient (A)
After purification by high performance liquid chromatography using (mobile phase), the product was treated with DowexlX2 (acetate type) and freeze-dried to obtain the title compound.

Yield 11: 27 mg R, 3: 0.14 [α]. : -154,0" (c=0.5.Water) FA
B mass spectrum (M+1): 676 [Example 6] 11-Pro-Cys-Pro-Arg-Gly-Nl
2-Acetate (1) Z-Pro-Cys(MBzl)-
Pro-Arg (Mbs)-Gly-Nl20oc-
C;ys(MBzl)-Pro-Arg(Mbs)-G
ly-NH□ 2.4g, 4NHCl-Ac0E
t 10mf1%NMM O, 6mfl and Z-Pro
From 1.3 g of -O5u, the title compound was obtained in the same manner as in Example 1-(4).

Yield: 2.3g Melting point: 101-104°C Rf': o, 41, Re': 0. 61
[α]o Knee 56.4° (c=1.0. DMF)
(2) it-Pro-Cys-Pro-Arg-Gl
y-Nl2・acetate Z-Pro-(:ys(MBzl)
-Pro-Arg(Mbs)-Gly-Nl-1215
0 mg of MSA in the same manner as in Example 1-(7).
- After anisole treatment, 12 ml 1 min (flow rate), 0
After high performance liquid chromatography purification in (A) (mobile phase) of 510% (B) 20 minute linear gradient, D
owexlx2 (acetate form) and freeze-drying to give the title compound.

Yield: 64 mg R13 (containing 0.1% ethanediol): 0.12 [αl
o knee 92.7° (c-0,5, water) FAB mass spectrum (M+1): 528 [Example 7] (H-Pro-Cys-Pro-^rg-Gly-NH
z )2 ・Acetate H-Pro-(:ys-Pro-A
30 mg of rg-Gly-Nl2-acetate was dissolved in 2 ml of water, and the pH was adjusted to 7 with dilute aqueous ammonia. After stirring at room temperature for 7 days, the mixture was acidified with acetic acid and freeze-dried.

Yield: 2Bmg R, 3:0.02 [α]o: 142,9” (c-0,5, water) FAB
Mass spectrum (M+1): 1054 Next, pharmacological test examples showing the effectiveness of the peptide derivative of the present invention will be shown.

[Pharmacological test example] The effect on memory consolidation was determined using Wistar male rats by Burbach et al.
Science), 2nd, 1310-1312 (1
This study was conducted using a trial passive avoidance experiment based on the method of [983]. The experimental setup consisted of a light room and a dark room, and the floor was made of stainless steel grids. Rats placed in the light room can freely move to the dark room. Using this device, rats are given a single electric shock when they enter a dark room.

Retention of passive avoidance behavior in response to electric shock was determined by the time it took for the rat, placed in the light room again after a certain period of time, to enter the dark room (response latency).

Examination of the effect of improving experimental retrograde amnesia by cycloheximide The peptide derivative of the present invention or physiological saline was administered subcutaneously, and 1 hour later, an electric shock (0.5 mA) was experienced, and immediately thereafter, cycloheximide was administered subcutaneously. A dose of 2.7 to 3.0 H/kg or physiological saline was administered subcutaneously, and a memory retention test was conducted 48 hours later. Rats to which only physiological saline was administered generally exhibited a response latency of around 300 seconds, whereas rats in the control group to which only cycloheximide was administered exhibited a response latency of approximately 50 seconds and developed retrograde amnesia.

The average response latency of the group administered with the peptide derivative of the present invention was compared with that of the control group. The number of rats used for testing each group is 6-8. The maximum measurement time was 600 seconds.

For the peptide derivatives obtained in each example, the dosage and effects (ratio of response latency on each side to response latency of the control group is shown in %) are shown in Table 1.

From the above test results in Table 1, it is clear that the peptide derivative of the present invention exhibits an excellent improvement effect on retrograde amnesia.

Next, examples of formulations of drugs containing the peptide derivatives of the present invention will be 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.
and adjust the pH to 6.0 to 8.0 with sodium hydroxide.
An aqueous solution was prepared. This was filtered for 1 m after bacterial filtration.
l1. The mixture was filled into ampoules, sealed and heat sterilized to produce an injection.

[Formulation Example 2] (Freeze-dried preparation) 5 mg of the peptide derivative obtained in Example 1 and 5 g of D-mannitol were contained in 100+nJZ of distilled water for injection, and the pH was adjusted to 6.0 to 6.0 with a phosphate buffer. An aqueous solution adjusted to 8.0 was prepared. This was subjected to bacterial filtration, dispensed in 1 ml portions into vials, and then freeze-dried to produce a freeze-dried injection.

[Formulation Example 3] (Nasal Drop) 10 mg of the peptide derivative obtained in Example 1 was contained in 100 mL of physiological saline, and the pH was adjusted to 3.0 to 6.0 with a citric acid buffer. A nasal spray containing 50 μg per 0.5 mL dose was produced.

[Formulation example 4 (planing) Hard fat (triglyceride of saturated fatty acids) 98
．． After adding 0.5 g of egg yolk lecithin to 5 g and melting at 40 to 45°C, the peptide derivative 5 obtained in Example 1 was added.
A solution obtained by dissolving 1 mg of PEG400 in 1 g of PEG400 was added thereto, stirred and dispersed, and then 1 g of the solution was poured into a planing mold, and after solidification, it was separated from the mold to produce a planing.

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

Patent applicant: Nippon Chemifa Co., Ltd. Patent applicant: Fujirebio Co., Ltd. Representative Patent Attorney Yasuo Yanagawa

Claims (1)

[Claims] 1. The following general formula (I): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, A is cyclopentylcarbonyl, Pro or p
Glu, B is Gly or β-Ala, W is a hydrogen atom or the following general formula (II): Y^1-Cys-Y^2 (II) (wherein Y^1 is H or CO- T and Y^2 is OH
or T (However, T is the following general formula (III): ▲ Numerical formula, chemical formula, table, etc. ▼ (III) (In the formula, R^1 is an alkylcarbonyl group having 2 to 7 carbon atoms, carbon A group selected from the group consisting of an arylcarbonyl group having 7 to 10 atoms and an alkylthio group having 1 to 6 carbon atoms), or the following general formula (IV): ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (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), or the following general formula (V): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(V) [In the formula, A and B are the above-mentioned ] A peptide derivative represented by the following, or a derivative in a functional group thereof, or a pharmacologically acceptable salt thereof. 2. The following general formula (I): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) [In the formula, A is cyclopentylcarbonyl, Pro or p
Glu, B is Gly or β-Ala, W is a hydrogen atom or the following general formula (II): Y^1-Cys-Y^2 (II) (wherein Y^1 is H or CO- T and Y^2 is OH
or T (However, T is the following general formula (III): ▲ Numerical formula, chemical formula, table, etc. ▼ (III) (In the formula, R^1 is an alkylcarbonyl group having 2 to 7 carbon atoms, carbon A group selected from the group consisting of an arylcarbonyl group having 7 to 10 atoms and an alkylthio group having 1 to 6 carbon atoms), or the following general formula (IV): ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (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), or the following general formula (V): ▲There are mathematical formulas, chemical formulas, tables, etc.▼(V) [In the formula, A and B are the above-mentioned or a functional group derivative thereof, or a pharmacologically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. An anti-dementia drug.