JPH0686378B2 - Immunomodulator - Google Patents

Description

TECHNICAL FIELD The present invention relates to immunomodulators.

(Prior Art) Conventionally, L-phenylalanyl-L-phenylalanyl-
As a peptide compound having an L-histidine methyl ester structure, for example, N-acetyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester is known to have an anti-ulcer effect (Japanese Patent Application Laid-Open No. 2004-242242). Sho 51
−136669).

(Problems to be Solved by the Invention) The present invention is different from the above-mentioned known compounds in which a terminal nitrogen atom is substituted with an acetyl group, on the nitrogen atom, a pivaloyl group
It is intended to provide an excellent immunomodulator containing a peptide into which a bulky group such as a tertiary butoxycarbonyl group and a benzyloxycarbonyl group is introduced as an active ingredient.

(Means for Solving the Problems) That is, the present invention relates to an immunomodulator comprising a novel imidazole-containing peptide represented by the following general formula or a pharmacologically acceptable salt thereof as an active ingredient.

(However, R ¹ is a branched alkyl group, a branched alkoxy group or an aryl group-substituted lower alkoxy group, R ² and R ⁴ are the same or different and are a hydrogen atom or a lower alkyl group, and R ³ and R ⁵ are phenyl group-substituted lower groups. An alkyl group, R ⁶ is a hydrogen atom or a lower alkoxycarbonyl group, R ⁷ is a hydrogen atom or a nitrogen-containing heterocyclic group-substituted lower alkylthio group, X ¹ and X ² are the same or different. A is a lower alkoxycarbonyl group, a hydroxymethyl group and a formula The lower alkylene group optionally having a substituent selected from the groups represented by, R ⁸ and R ⁹ are the same or different and represent a hydrogen atom or a lower alkyl group. The imidazole-containing peptide (I) and its pharmacologically acceptable salt, which are the active ingredients of the present invention, show remarkable macrophage migration promoting activity in the macrophage migration test used for measuring cellular immune activity. Also,
The imidazole-containing peptide (I) exerts an excellent immunomodulatory effect of suppressing delayed allergic reaction.
In addition, the imidazole-containing peptide (I) or a pharmacologically acceptable salt thereof has low toxicity and shows high safety as a medicine. For example, 4- [2- (3-tert-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanyl) aminoethyl] -5-methoxycarbonyl-2- (2-pyridylmethylthio) imidazole is orally administered to mice. After observation (dose of 300 mg / kg) for 3 days, no deaths were observed.

In the imidazole-containing peptide (I) which is the active ingredient of the present invention, specific examples of the compound in which R ¹ is an aryl group include compounds in which R ¹ is a phenyl group and R ⁷ is a nitrogen-containing heterocyclic group. , R ⁷ is a pyridyl group, respectively. Further, the peptide (I) is preferably one in which R ⁶ , R ⁷ and group A are substituted at the 5-position, 2-position and 4-position of the imidazolyl group, respectively, and further, carbon atoms of a branched alkyl group and a branched alkoxy group. It is preferable that the number is 3 to 6, and the lower alkyl group, the lower alkoxy group and the lower alkylene group have 1 to 4 carbon atoms.

A therapeutically more preferred compound is that R ¹ is a tertiary butyl group, a tertiary butoxy group or a benzyloxy group, and R ² and R ⁴
Is a hydrogen atom or a methyl group, R ³ and R ⁵ are benzyl groups or phenethyl groups, R ⁶ is a hydrogen atom or a methoxycarbonyl group, R ⁷ is a hydrogen atom or a 2-pyridylmethylthio group, A Is a methylene group, ethylene group, methoxycarbonylethylene group, hydroxymethylethylene group or carbamoylethylene group.

The imidazole-containing peptide (I) can have the tautomeric structure shown below in the imidazole moiety, but the active ingredient of the present invention includes both isomers.

(However, the symbols have the same meanings as described above.) In addition, the active ingredient compound (I) of the present invention includes both an optical isomer and a racemate when it has an asymmetric carbon atom in the molecule. Especially preferred are compounds in which all the asymmetric carbon atoms are in the S configuration.

The imidazole-containing peptide (I) which is the active ingredient of the present invention can be used for the purpose of the present invention in a free form or in the form of a pharmaceutically acceptable salt thereof. Such pharmacologically acceptable salts include, for example, inorganic acid addition salts such as hydrochloride, hydrobromide, phosphate and sulfate,
Alternatively, organic acid addition salts such as oxalate, acetate, lactate, citrate, tartrate, fumarate, maleate, aspartate, methanesulfonate and benzoate may be mentioned.

The immunomodulator of the present invention can be administered orally or parenterally, and can be used as an appropriate pharmaceutical preparation such as tablets, granules, capsules, powders and injections by a conventional method. You can

The dose of the immunomodulator of the present invention varies depending on the administration method, age, weight, condition of patient and type of disease to be treated, but the dose of the imidazole-containing peptide (I) or a salt thereof as an active ingredient is , Usually about 0 per day.
It is preferably administered at a dose of 01 to 100 mg / kg, especially 0.1 to 30 mg / kg.

The imidazole-containing peptide (I), which is the active ingredient of the present invention, can be prepared by (However, the symbols have the same meanings as described above.) The compound, salt or reactive derivative thereof and the general formula (However, the symbols have the same meanings as described above) or by subjecting the compound or a salt thereof to a condensation reaction, or (B) the general formula (However, the symbols have the same meanings as described above.) The compound, salt or reactive derivative thereof and the general formula (Where the symbols have the same meanings as described above), or a compound or salt thereof is subjected to a condensation reaction, or (C) the general formula R ¹ COOH (VI) (where the symbols have the same meanings as described above). A compound represented by the formula (1), a salt thereof or a reactive derivative thereof and a general formula (However, the symbols have the same meanings as described above.) The compound or salt thereof can be subjected to a condensation reaction to produce the compound.

Any of these condensation reactions can be carried out according to a conventional method for peptide synthesis. For example, compound (II),
When (IV) or (VI) is used in the form of its reactive derivative, the condensation reaction can be carried out in the presence or absence of a conventional deoxidizing agent (eg trialkylamine).

On the other hand, when the compound (II), (IV) or (VI) is used in the form of a free carboxylic acid or its salt, the condensation reaction can be carried out in the presence of a conventional condensing agent (eg dicyclohexylcarbodiimide). .

(Operation) Experimental Example 1 (Alveolar macrophage migration promoting activity) Japanese white rabbits (female, body weight 3 to 4 kg) were killed by bleeding under anesthesia, and lungs were washed with physiological saline to collect alveolar macrophages. The alveolar macrophages obtained were
The cells were allowed to migrate at 37 ° C for 24 hours in RPMI-1640 medium containing 5% rabbit serum containing 10 ^-7 M sample. The migration area was magnified and seen through, and the migration area was measured.

Also, RPM containing 5% rabbit serum containing 5 mM L-fucose
The migration area when allowed to migrate in the I-1640 medium was used as a control group. The alveolar macrophage migration promoting activity was determined using the migration index calculated by the following formula as an index.

As a result, all the compounds shown in Table 1 below have a migration index of 100.
The above is shown.

Experimental Example 2 (Picryl chloride delayed-type skin reaction inhibitory effect) 1 ml of an ethanol solution of picryl chloride (7w / v) was applied to the abdominal skin of BALB / c female mice (10 weeks old) for sensitization. The test compound was orally administered once daily for 7 days from the day of sensitization. Seven days after the sensitization, 10 μl of a 1 W / V% picryl chloride oriep oil solution (1 w / v%) was applied to both sides of the left ear skin, 5 μl each. Twenty-four hours after that, the mouse was bled to death and the thickness of both ears was measured.

The delayed skin reaction inhibitory effect was calculated as the ear swelling inhibitory rate (%) calculated by the following formula.

The results are as shown in Table 2 below.

Experimental Example 3 (Immunoregulatory action on spontaneously autoimmune disease animals) [Method] MRL / l mice (female, 5 weeks old, 8 mice per group) were subjected to 1 sample per day
Oral administration was performed once daily for 15 weeks. From the 7th week after the start of oral administration, the appearance of the mice was observed every week to count the number of swellings in the left and right dorsal lymph nodes. Also, urine was collected by bladder compression at 3, 5, 7, 9 and 11 to 13 weeks after the start of oral administration, and the protein amount was measured by Bio
-Quantified using RAD protein assay kit. further,
At 15 weeks after the start of oral administration, the animals were dissected and the weights of axillary lymph nodes and spleen were measured.

[Sample compound]

The samples used in this experiment are the compounds shown in Table 3 below.

[Results] 1. Survival rate The survival rate at 15 weeks (20 weeks of age) after the start of oral administration is
It is as shown in the table.

2. Inhibition rate of lymph node swelling The inhibition rate at 15 weeks (20 weeks of age) after oral administration is 5
It is as shown in the table.

3. Suppression rate of urinary protein concentration The suppression rate at 13 weeks (18 weeks old) after the start of oral administration was calculated by the following formula. The results are shown in Table 6 below.

For sample non-administration group 4. Inhibition rate of axillary lymph node weight increase The inhibition rate at 15 weeks (20 weeks old) after oral administration was calculated by the following formula. The results are shown in Table 7 below.

For sample non-administration group 5. Inhibition rate of increase in spleen weight The inhibition rate at 15 weeks (20 weeks of age) after the start of oral administration was calculated by the following formula. The results are shown in Table 8 below.

For sample non-administration group (Production Example) Production Example 1 L-phenylalanyl-L-histidine methyl ester dihydrobromide (1.75 g) was added to dimethylformamide (1
0 ml) and triethylamine (1.4 ml), N-te
Add rt-butoxycarbonyl-L-phenylalanine succinimide ester (1.34 g) and stir at room temperature overnight. Ethyl acetate was added to the reaction mixture, the insoluble material was filtered off, the filtrate was washed, and brine was added. The precipitated crystals are washed and dried to obtain N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester (1.5 g) as colorless crystals.

Yield: 71.8% mp.170-173 ° C (decomposition) Production Example 2 N-tert-butoxycarbonyl-L-2-amino-4-
After dissolving phenylbutyric acid / dicyclohexylamine salt (1.84 g) in ethyl acetate and 3% potassium hydrogen sulfate aqueous solution, the ethyl acetate layer is separated, washed with water and dried, and then the solvent is distilled off. The residue was dissolved in dimethylformamide (20 ml), L-phenylalanyl-L-histidine methyl ester dihydrobromide (1.91 g), N-hydroxysuccinimide (0.5 g) and dicyclohexylcarbodiimide (0.9 g). g), triethylamine (1.4 ml) is added while cooling, and the mixture is stirred at room temperature overnight. After the reaction, add ethyl acetate,
The insoluble material was removed by filtration, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform: methanol = 9: 1), crystallized from ethyl acetate and N-tert-
Butoxycarbonyl-L-2-amino-4-phenylbutyryl-L-phenylalanyl-L-histidine methyl ester (1.45 g) is obtained.

Yield: 61% mp.186-187 ° C (decomposition) Production Example 3 (1) N-tert-butoxycarbonyl-L-2-amino-4-phenylbutyric acid (2.79 g), L-histidine methyl ester dihydrochloride Salt (2.42g), 1-Hydroxybenzotriazole (1.35g) and triethylamine (2.8ml)
Is dissolved in dimethylformamide (20 ml), dicyclohexylcarbodiimide (2.1 g) is added under ice cooling, and the mixture is stirred at room temperature overnight. Ethyl acetate is added to the reaction solution, the insoluble matter is filtered off, the filtrate is washed and dried, and then the solvent is distilled off. The residue is crystallized with ether to give N- [N-tert-butoxycarbonyl-L-2-amino-4-phenylbutyryl] -L-histidine mer ester (3.3 g) as white crystals.

Yield: 76.6% mp.127-130 ° C (2) This product (2.15 g) is dissolved in 20 ml of 20% hydrobromide-acetic acid and stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, the residue was solidified with ether, and N- (L-2-amino-4-phenylbutyryl) -L-histidine methyl ester.
The hydrobromide salt is obtained. Use this product as dimethylformamide (20
ml), N-tert-butoxycarbonyl-L-phenylalanine succinimide ester (1.81 g) and triethylamine (2.1 ml) are added, and the mixture is stirred overnight. The reaction solution was treated in the same manner as in Production Example 2, crystallized with ether,
N- (N-tert-butoxycarbonyl-L-phenylcalanyl) -L-2-amino-4-phenylbutyryl-L
-Histidine methyl ester (3.3g) is obtained as white crystals.

Yield: 69.2% mp, 186 to 189 ° C Production Example 4 (1) N-benzyloxycarbonyl-L-phenylalanine (2.99g) and p-nitrophenol (1.39g) were dissolved in tetrahydrofuran (30ml) and cooled under ice. Add dicyclohexylcarbodiimide (2.1 g) and stir for 4 hours. The insoluble matter was removed by filtration, ethyl acetate was added to the filtrate, and the filtrate was washed,
After drying, the solvent is distilled off. Hexane is added to the residue to obtain N-benzyloxycarbonyl-L-phenylalanine p-nitrophenyl ester as crystals. This product is dissolved in tetrahydrofuran (20 ml), L-histidine methyl ester dihydrochloride (2.4 g) and triethylamine (2.8 ml) in water (15 ml) are added under ice-cooling, and the mixture is stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, the residue was purified by silica gel column chromatography (chloroform: methanol = 15: 1), and crystallized from ethyl acetate.
N-benzyloxycarbonyl-L-phenylalanyl-L-histidine methyl ester (2.8 g) is obtained as colorless crystals.

Yield: 64.1% mp.113-116 ° C (2) This product (2.49 g) is dissolved in 25% hydrogen bromide-acetic acid (30 ml) and stirred at room temperature for 1 hour. The reaction solution is concentrated under reduced pressure,
The residue is solidified with ether to give crude L-phenylalanyl-L-histidine methyl ester dihydrobromide. A mixture of this product (1.75 g), N-benzyloxycarbonyl-L-phenylalanine, p-nitrophenyl ester (1.55 g), triethylamine (1.4 ml) and dimethylformamide (10 ml) was stirred overnight at room temperature, and the reaction mixture was stirred. Concentrate under reduced pressure, add ethyl acetate and water to the residue,
The precipitated crystals were collected by filtration to give N-benzyloxycarbonyl-L.
-Phenylalanyl-L-phenylalanyl-L-histidine methyl ester (1.02g) is obtained.

Yield: 46.2% mp.204-206 ° C Production Example 5 4- [2- (N-benzyloxycarbonylamino) ethyl] -5-methoxycarbonylnimidazole (1.2 g)
Dissolve in 30% hydrogen bromide-acetic acid solution (20 ml) and stir at room temperature for 30 minutes. The reaction solution is concentrated under reduced pressure, and ether is added to the residue to solidify it to obtain 4- (2-aminoethyl) -5-methoxycarbonylimidazole dihydrobromide. Dimethylformamide (20 ml), N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanine (1.65 g) and 1-hydroxybezotriazole (0.54 g) were added to this product. Dicyclohexylcarbodiimide (0.83 g) was added under ice-cooling, the mixture was stirred for 10 minutes, triethylamine (1.4 ml) was added, and the mixture was stirred at room temperature overnight. Ethyl acetate was added to the reaction mixture, the insoluble material was filtered off, the filtrate was washed,
After drying, the solvent is distilled off. The residue was purified by silica gel column chromatography (chloroform: methanol = 12: 1) and crystallized from ether to give 4- {2- [N- (N
-Tert-butoxycarbonyl-L-phenylalanyl-
L-Phenylalanyl) amino] ethyl] -5-methoxycarbonylimidazole (1.4 g) is obtained as colorless crystals.

Yield: 62.2% mp.155-159 ° C Production Examples 6-8 N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanine or N-benzyloxycarbonyl-L-phenylalanyl-L-phenylalanine, 4-aminomethyl-5-methoxycarbonyl-2-
(2-pyridylmethylthio) imidazole or 4- (2
-Aminoethyl) -5-methoxycarbonyl-2- (2
-Pyridylmethylthio) imidazole is treated as in Preparation Example 5 to give the following compound:

(6) 4- [N- (N-tert-butoxycarbonyl-
L-phenylalanyl-L-phenylalanyl) aminomethyl] -5-methoxycarbonyl-2- (2-pyridylmethylthio) imidazole mp.147-150 ° C (7) 4- {2- [N- (N- tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl)
Amino] ethyl} -5-methoxycarbonyl-2- (2
-Pyridylmethylthio) imidazole mp.182-184 ° C (decomposition) (8) 4- [N- (N-benzyloxycarbonyl-
L-phenylalanyl-L-phenylalanyl) aminomethyl] -5-methoxycarbonyl-2- (2-pyridinemethylthio) imidazole mp.140-142 ° C Production Example 9 (1) Histamine dihydrochloride (1.84 g) ), N-tert-butoxycarbonyl-L-phenylalanine succinimide ester (3.62 g), dimethylformamide (20 ml)
A mixture of water and water (5 ml) under ice-cooling triethylamine (3.5
ml) and stirred at room temperature for 3 hours. The reaction mixture is concentrated under reduced pressure, ethyl acetate is added to the residue, and the insoluble material is filtered off.
After washing and drying the filtrate, the solvent is distilled off. The residue was crystallized from ether to give N- (N-tert-butoxycarbonyl-
L-phenylalanyl) histamine (2.5 g) is obtained. This product (1.79 g) is dissolved in a 15% hydrogen bromide-acetic acid solution and stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure to give N- (L-phenylalanyl) histamine dihydrobromide (2.1 g).
To get This product (2.1 g) is dissolved in dimethylformamide, N-tert-butoxycarbonyl-L-phenylalanine succinimide ester (1.81 g) and triethylamine (2.1 ml) are added, and the mixture is stirred overnight. Ethyl acetate is added to the reaction mixture, the insoluble matter is filtered off, the filtrate is washed and dried, and then the solvent is distilled off. The residue was purified by silica gel chromatography (chloroform: methanol = 9: 1) and crystallized from ether to give N- (N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl) histamine (1.88 g). ) As white crystals.

Yield: 74.3% mp.175-177 ° C Production Example 10 N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester (2.25 g) in 10% hydrogen chloride-dioxane solution (30 ml)
And stir at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, ether was added to the residue, and L-phenylalanyl-
L-Phenylalanyl-L-histidine methyl ester dihydrochloride (2.15 g) is obtained as a white powder. This product (2.1
g) is dissolved in dimethylformamide (15 ml), pivaloyl chloride (0.48 g) is added, triethylamine (2.24 ml) is added dropwise under ice cooling, and the mixture is stirred at room temperature overnight. Ethyl acetate is added to the reaction mixture, and the insoluble material is filtered off. The filtrate is washed, the dried liquid and the solvent are distilled off. The residue was purified by silica gel chromatography (chloroform: methanol = 12: 1) and then solidified with ether to give N-pivaloyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester (0.98 g). obtain.

Yield: 44.8% mp.128-131 ° C Production Example 11 N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester (2 g) was dissolved in ammonia saturated ethanol solution, Stir in a pressure bottle at room temperature for 3 days. The reaction mixture is concentrated under reduced pressure, and the residue is crystallized with ether to give N-tert-butoxycarbonyl-L-phenylalanyl-L-phenylalanyl-L-histidine amide (1.7 g) as white crystals.

Yield: 87.2% mp.169-171 ° C (decomposition) Production Example 12 N-pivaloyl-L-phenylalanyl-L-phenylalanyl-L-histidine methyl ester (1.90 g) was dissolved in tetrahydrofuran (12 ml), A solution of sodium borohydride (0.20 g) in methanol (2.5 ml) was added dropwise under ice cooling. After stirring at room temperature for 4 hours, 5% hydrochloric acid (4 ml) is added and the solvent is evaporated. The mixture is made alkaline with aqueous sodium hydrogen carbonate and extracted with ethyl acetate. After the extract is washed and dried, the solvent is distilled off. The residue was solidified with ethyl acetate and 4
-{2-Hydroxymethyl-2- [N- (N-pivaloyl-L-phenylalanyl-L-phenylalanyl) amino] ethyl} imidazole (1.24 g) is obtained as a white powder.

Yield 68.8% mp.128-130 ° C Production Example 13 (1) L-phenylalanine methyl ester hydrochloride (4.3g), triethylamine (6.4ml) was added to a solution of fosge (2.6g) in dichloromethane (25ml) at -30 ° C or lower. And a mixture of dichloromethane (25 ml) is added dropwise. After stirring at -30 ° C for 30 minutes, the reaction mixture is concentrated under reduced pressure. Dimethylformamide (30 ml), N-tert-butoxycarbonyl-N 'was added to the residue.
-Benzylhydrazine (4.44 g) and triethylamine (3.64 ml) are added, and the mixture is stirred at 50 ° C for 5 hours and further at room temperature overnight. The reaction mixture is concentrated under reduced pressure and ethyl acetate and water are added to the residue. The organic layer is separated, washed with water and dried, and then the solvent is distilled off. The residue was purified by silica gel chromatography (toluene: ethyl acetate = 8: 1) and solidified with hexane to give 3-tert-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanine methyl ester (4g).
As a white powder.

Yield: 93.5% mp.71-73 ℃ (2) Dissolve this product (1.71g) in methanol (5ml) and add 2N.
-Add aqueous sodium hydroxide (2.2 ml) and stir at room temperature for 3 hours. After the reaction, remove methanol under reduced pressure, acidify with 10% citric acid, and extract with ethyl acetate. The extract is washed and dried, and the solvent is evaporated to give 3-tet-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanine (1.65 g) as crude crystals. This product (1.65g) was dissolved in dimethylformamide (20ml) and histidine methyl ester dihydrochloride (1.07g), triethylamine (1.26m) was added.
l), 1-hydroxybenzotriazole (0.54 g) and dicyclohexylcarbodiimide (0.91 g) are added under ice cooling, and the mixture is stirred at room temperature overnight. Ethyl acetate is added to the reaction mixture, and the insoluble material is filtered off. After washing and drying the filtrate, the solvent is distilled off. The residue was purified by silica gel chromatography (chloroform: methanol = 12: 1) and then solidified with ether to give 3-tert-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanyl-L-histidine methyl ester (1.76). g) is obtained as a colorless powder.

Yield: 77.9% NMR (CDCl ₃ ) δ: 1.40 (9H, s) 2.9-3.3 (4H, m), 3.65
(3H, s), 4.4-5.0 (4H, m), 6.07 (1H, d), 6.65-6.9
(1H, m), 6.70 (1H, s), 7.0−7.5 (1H, m), 7.39 (1H, m
m) Production Examples 14-15 3-tert-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanine and histamine dihydrochloride or 4- (2-aminoethyl) -5-methoxycarbonyl-2- (2-pyridyl) Methylthio) imidazole trihydrobromide is treated in the same manner as in Production Example 13- (2) to give the following compound.

(14) N- (3-tert-butoxycarbonyl-2-benzylcarbazoyl-L-phenylalanyl) histamine NMR (CDCl ₃ ) δ: 1.35 (9H, s), 2.5-2.8 (2H, m), 2.9-
3.5 (4H, m), 4.3-4.8 (3H, m), 6.03 (1H, brd), 6.63
(1H, s), 6.9-7.4 (12H, m) (15) 4- [2- (3-tert-butoxycarbonyl-
2-benzylcarbazoyl-L-phenylalanyl) aminoethyl] -5-methoxycarbonyl-2- (2-pyridylmethylthio) imidazole mp.132-137 ° C Production Examples 16-18 L-phenylalanine methyl ester and N-tert. -Butoxycarbonyl-N'-phenethylhydrazine or N-
Pivaloyl-N′-bedylhydrazide hydrobromide,
The following compound is obtained by treating in the same manner as in Production Example 13- (1).

3-tert-butoxycarbonyl-2-phenethylcarbazoyl-L-phenylalanine methyl ester mp.97-99 ° C 3-pivaloyl-2-benzylcarbazoyl-L-phenylalanine methyl ester mp.141-143 ° C obtained above Compound and histamine dihydrochloride or 4- (2
-Aminoethyl) -5-methoxycarbonyl-2- (2
-Pyridylmethylthio) imidazole trihydrobromide is treated in the same manner as in Production Example 13- (2) to obtain the following compound.

(16) N- [3-tert-butoxycarbonyl-2-phenethylcarbazoyl-L-phenylalanyl] histamine mp.87-95 ° C (17) N- (3-pivaloyl-2-bedylcarbazoyl-L -Phenylalanyl) histamine mp.78-86 ° C (18) 5-methoxycarbonyl-4- [2- (3-pivaloyl-2-benzylcarbazoyl-L-phenylalanyl) aminoethyl] -2- (2 -Pyridylmethylthio) imidazole mp.171-172 ° C Production Example 19 (1) N- (benzyloxycarbonyl) -L-2-aminooxy-3-phenylpropionic acid (3.09g), L
-Histidine methyl ester dihydrochloride (2.42g), N
-Hydroxybenzotriazole (1.35g), triethylamine (2.8ml) and dimethylformamide (20ml)
Of ice-cooled sidiclohexyl cardiimide (2.1
g) and stir overnight at room temperature. The reaction mixture is concentrated under reduced pressure, ethyl acetate is added to the residue, the insoluble matter is filtered off, the filtrate is washed and dried, and then the solvent is distilled off. The residue was subjected to silica gel column chromatography (chloroform: ethyl acetate =
12: 1) and solidified with hexane to give N- [N- (benzyloxycarbonyl) -L-2-aminooxy-3
-Phenylpropionyl] -L-histidine methyl ester (2.1 g) is obtained as a white powder.

NMR (CDCl ₃ ) δ: 2.8-3.2 (4H, m), 3.62 (3H, s), 4.3-
4.9 (2H, m), 5.09 (2H, s), 6.66 (1H, s), 7.0-7.5 (10
H, m), 7.36 (1H, s), 7.95 (1H, d) (2) Dissolve this product (2.1g) in 33% hydrogen bromide-acetic acid (20ml), stir at room temperature for 1 hour, and then reduce the pressure. Concentrate. Ether was added to the residue to solidify it, and N- (L-2-aminooxy-
3-Phenylpropionyl) -L-histidine methyl ester dihydrobromide (2.32 g) is obtained. This product (2.32
g) was suspended in dimethylformamide (20 ml), N-t
ert-butoxycarbonyl-L-phenylalini (1.25
g), 1-hydroxybezotriazole (0.68 g) and triethylamine (1.3 ml) are added, and dicyclohexylcarbodiimide (1.1 g) is further added under ice cooling, and the mixture is stirred overnight at room temperature. The reaction solution is concentrated under reduced pressure, ethyl acetate is added to the residue, the insoluble matter is filtered off, the filtrate is washed and dried, and then the solvent is distilled off. The residue was purified by silica gel column chromatography (chloroform: methanol = 15: 1), solidified with isopropyl ether, and N- [N- (tert-butoxycarbonyl-L-phenylalanyl) -L-2-aminooxy was used. -3-Phenylpropionyl] -L-histidine methyl ester (1.95g) is obtained as a white powder.

Yield: 71.7% NMR (CDCl ₃ ) δ: 1.35 (9H, s), 2.7-3.4 (6H, m), 3.70
(3H, s), 4.0-5.0 (3H, m), 5.0-5.5 (1H, m, 6.74 (1
H, s), 7.0-8.3 (11H, d) Production Example 20 N- (benzyloxycarbonyl) -L-2-aminooxy-3-phenylpropionic acid (1.6 g), L-phenylalanyl-L-histidine Methyl ester dihydrobromide (2.39g), N-hydroxysuccinimide (0.6
A mixture of g), triethylamine (1.4 ml) and dimethylformamide (20 ml) was added with dicyclohexylcarbodiimide (1.1 g) under ice cooling, and the mixture was stirred at room temperature overnight. The reaction solution is concentrated under reduced pressure, ethyl acetate is added to the residue, the insoluble matter is filtered off, the filtrate is washed and dried, and then the solvent is distilled off. The residue was purified by silica gel column chromatography (chloroform: methanol = 9: 1), crystallized from ether, and N-
[N- (benzyloxycarbonyl) -L-2-aminooxy-3-phenylpropionyl] -L-phenylalanyl-L-histidine methyl ester (1.3 g) is obtained.

NMR (CDCl ₃ ) δ: 2.6-3.3 (6H, m), 3.62 (2H, s) 4.3-
4.9 (3H, m), 5.05 (2H, s), 6.63 (1H, s), 6.8−7.9 (17
H, m), Production Example 21 N-pivaloyl-L-phenylalanine and N-methyl-
L-Phenylalanyl-L-histidimethyl ester was treated as in Preparation Example 1 to give N-pivaloyl-L-phenylalanyl-L-N-methyl-phenylalanyl-L-.
Histidine methyl ester is obtained as a colorless powder.

Production Example 22 N-pivaloyl-L-N-methyl-phenylalanine and L-N-methyl-phenylalanyl-L-histidine methyl ester were treated in the same manner as in Production Example 1 to give N-pivaloyl-N-methyl-L-. Phenylalanyl-L-N-methyl-phenylalanyl-L-histidine methyl ester is obtained.

Production Example 23 N- (N-pivaloyl-L-phenylalanyl) -N '
-Benzylhydrazine and L-histidine methyl ester are treated in the same manner as in Production Example 13 to obtain N-pivaloyl-L-phenylalanyl-2-benzylcarbazoyl-L-histidine methyl ester as a colorless powder.

Production Example 24 N- (3-pivaloyl-2-benzylcarbazoyl)-
N'-Benzylhydrazine and L-histidine methyl ester are treated as in Preparation 13 to give 3- (3-pivaloyl-2-benzylcarbazoyl) -2-benzylcarbazoyl-L-histidine methyl ester.

Production Example 25 N-pivaloyl-L-phenylalanine and L-2-aminooxy-3-phenylpropionyl-L-histidine methyl ester were treated in the same manner as in Production Example 19 to give N- (N
-Pivaloyl-L-phenylalanyl) -L-2-aminooxy-3-phenylpropionyl-L-histidine ethyl ester is obtained.

Production Example 26 Production Example 19 of N-pivaloyl-L-2-aminooxy-3-phenylpropionic acid and L-2-aminooxy-3-phenylpropionyl-L-histidine methyl ester
Is treated in the same manner as in to obtain N- (N-pivaloyl-L-2-aminooxy-3-phenylpropionyl) -L-2-aminooxy-3-phenylpropionyl-L-histidimethyl ester.

Production Example 27 N-pivaloyl-L-N-methyl-phenylalanine and L-phenylalanyl-L-histidine methyl ester were treated in the same manner as in Production Example 1 to give N-pivaloyl-L-N.
-Methyl-phenylalanyl-L-phenylalanyl-
L-histidine methyl ester is obtained as a colorless powder.

Production Example 28 N-pivaloyl-N'-benzylhydrazine and N- (L
2-Aminooxy-3-phenylpropionyl) histamine was treated in the same manner as in Production Example 13 to give N- (3-pivaloyl-2-benzylcarbazoyl-L-2-aminooxy-3-phenylpropionylhistamine as colorless Obtained as a powder.

Production Example 29 N-pivaloyl-L-N-methyl-phenylalanyl-
L-Phenylalanyl-L-histidine methyl ester was treated as in Preparation 11 to give N-pivaloyl-LN.
-Methyl-phenylalanyl-L-phenylalanyl-
L-histidine amide is obtained as a colorless powder.

Production Examples 30 to 35 The compounds obtained in Production Examples 21 to 26 are amidated in the same manner as in Production Example 11 to obtain the compounds shown in Table 9 below.

(Effects of the Invention) The imidazole-containing peptide (I) and its pharmacologically acceptable salt, which are the active ingredients of the present invention, have a macrophage migration promoting activity, a delayed allergic reaction inhibitory effect, and a prolongation of life in animals with autoimmune disease. Since it has the effect of suppressing swelling of lymph nodes, suppressing protein in urine, and suppressing increase in weight of axillary lymph nodes and spleen, it enhances immune activity when immune is lowered and suppresses it when immune response is increased It has an excellent immunomodulatory effect of recovering the level. Based on such an immunomodulatory action, the immunomodulator of the present invention is a cellular arterial such as lithium arthritis, multiple sclerosis, systemic lupus erythematosus, glomerular sickness, rheumatic fever, type I diabetes and atopic allergies. It can be used as a therapeutic and / or preventive agent for immune diseases and the like.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display location A61K 37/02 ACV 8314-4C ADP 8314-4C // C07D 233/64 106 233/66 233/84 C07K 5/06 5/08 (56) Reference JP-A-2-262561 (JP, A)

Claims (5)

[Claims] 1. A general formula (However, R ¹ is a branched alkyl group, a branched alkoxy group or an aryl group-substituted lower alkoxy group, R ² and R ⁴ are the same or different and are a hydrogen atom or a lower alkyl group, and R ³ and R ⁵ are phenyl group-substituted lower groups. An alkyl group, R ⁶ is a hydrogen atom or a lower alkoxycarbonyl group, R ⁷ is a hydrogen atom or a nitrogen-containing heterocyclic group-substituted lower alkylthio group, X ¹ and X ² are the same or different. A is a lower alkoxycarbonyl group, a hydroxymethyl group and a formula The lower alkylene group optionally having a substituent selected from the groups represented by, R ⁸ and R ⁹ are the same or different and represent a hydrogen atom or a lower alkyl group. ) An immunomodulator comprising the imidazole-containing peptide represented by or a pharmacologically acceptable salt thereof as an active ingredient. 2. R ¹ is a branched alkyl (C _3-6 ) group, a branched alkoxy (C _3-6 ) group or a phenyl group-substituted alkoxy (C
The immunomodulator according to claim 1, which is a _1-4 ) group, and R ⁷ is a hydrogen atom or a pyridyl group-substituted alkyl (C _1-4 ) thio group. (3) The immunomodulator according to (1) or (2), wherein the group A is bonded to the 4-position of the imidazolyl group. Wherein R ¹ is tertiary butyl, tertiary butoxy or benzyloxy group, R ² and R ⁴ is a hydrogen atom or a methyl group, R ³ and R ⁵ are benzyl or phenethyl group, R ⁶ Is a hydrogen atom or a methoxycarbonyl group, R ⁷ is a hydrogen atom or 2-
Pyridylmethylthio group, A is methylene group, ethylene group,
A methoxycarbonylethylene group, a hydroxymethylethylene group or a carbamoylethylene group (3).
The described immunomodulator. 5. Lithium arthritis, multiple sclerosis, systemic lupus erythematosus, glomerular inflammation, rheumatic fever, type I diabetes,
The immunomodulator according to any one of claims (1) to (4), which is a therapeutic and / or preventive agent for atopic allergy.