JPH0616625A - Amino acid derivative - Google Patents

Abstract

PURPOSE:To provide a new amino acid derivative which is useful as an immunoactivating agent against tumors, bacterium infections or autoimmune diseases and as a host-controlled carcinostatic agent. CONSTITUTION:The compound of formula I [R1 is benzyl, phenyl which may be substituted (may be substituted with 1 to 3 halogen atoms), X is -CO-, -SO2-; Z is methylene, S; R<3> is H; R<4> is H, 1 to 4C alkyl; R<5> is 1 to 4C alkyl; R<6> is phenyl], for example, a compound of formula I where R<1>, R<6> is phenyl, X is CO, Z is CH2, R<3> is H, R<4> and R<5> are methyl. The compound of formula I is obtained by allowing a formula of formula II to react with a compound of formula III in the presence of a condensing agent, or its reactive derivative followed by reaction of the product of formula IV with a compound of formula V.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an amino acid derivative useful as an immunostimulant and a carcinostatic agent.

[0002]

2. Description of the Related Art Although synthetic examples of pyrrolidine or thiazolidine derivatives have been known, little is known about the pharmacological action of these compounds.

[0003]

The present inventors have found that various pyrrolidine or thiazolidine derivatives have excellent immunostimulating action and antitumor action, and completed the present invention.

[0004]

The present invention has the general formula

[0005]

[Chemical 2]

[In the formula, R ¹ is a benzyl group, a phenyl group which may have a substituent (the substituent represents a halogen atom (fluorine, chlorine, bromine)), and these substituents are 1 to 3
You may have one. ), X is a -CO- group or -S
O ₂ - represents a group, Z is represents a methylene group or a sulfur atom, R ³ represents a hydrogen atom, R ⁴ is a hydrogen atom or an alkyl group (methyl having 1 to 4 carbon atoms, ethyl, propyl,
Isopropyl or butyl, etc., and R ⁵ is an alkyl group having 1 to 4 carbon atoms (methyl, ethyl, propyl,
Isopropyl, butyl, etc.) and R ⁶ represents a phenyl group. ] It is a compound having.

The compound having the general formula (I) can be obtained by the following two methods (A) and (B).

Method A General formula

[0009]

Embedded image A compound having R ¹ XOH (II) (wherein R ¹ and X have the same meanings as defined above) in the presence of a condensing agent or a reactive derivative thereof and the general formula

[0010]

[Chemical 4]

A compound having the general formula (wherein Z has the same meaning as described above) is reacted.

[0012]

[Chemical 5]

(Wherein R ¹ , X and Z have the same meanings as defined above), the compound of the general formula

[0014]

Embedded image A compound having HR ³ NCR ⁴ R ⁵ R ⁶ (V) (wherein R ³ , R ⁴ , R ⁵ and R ⁶ have the same meanings as described above) in the presence of a condensing agent. General formula when reacted

[0015]

[Chemical 7]

(Wherein R ¹ , X, Z, R ³ , R ⁴ , R ⁵
And R ⁶ have the same meaning as described above. ) Are obtained.

Method B General formula

[0018]

[Chemical 8]

(Wherein Z has the same meaning as defined above) and a general formula

[0020]

[Image Omitted] A compound having HR ³ NCR ⁴ R ⁵ R ⁶ (V) (wherein R ³ , R ⁴ , R ⁵ and R ⁶ have the same meanings as described above) is condensed to give a compound of the general formula

[0021]

[Chemical 10]

(Wherein Z, R ³ , R ⁴ , R ⁵ and R ⁶
Has the same meaning as described above. ) To a compound having the general formula

[0023]

Embedded image When a compound having R ¹ XOH (II) (wherein R ¹ and X have the same meanings as described above) or a reactive derivative thereof is reacted,

[0024]

[Chemical 12]

(Wherein R ¹ , X, Z, R ³ , R ⁴ , R ⁵
And R ⁶ have the same meaning as described above. ) Are obtained.

In Method A and Method B, the reactive derivative of the general formula (II) is an acid halide or a sulfonyl halide (wherein halogen is a chlorine or bromine atom) or an acid anhydride. .

When the acid halide or sulfonyl halide of (II) is used in the reaction of (II) and (III) in Method A, a base (sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, triethylamine, pyridine, or The reaction is performed in the presence of (for example, picoline).

The solvent used in the reaction is water, methanol, dioxane, methyl cellosolve, methylene chloride, chloroform, benzene, toluene, dimethylformamide or dimethylacetamide.

The reaction temperature is 0 ° C to 100 ° C, and the reaction time is 30 minutes to 2 hours.

The compound having the general formula (IV) obtained by this reaction is purified by a conventional method.

The condensation reaction between (IV) and (V) is carried out by using a condensing agent (dicyclohexylcarbodiimide, carbonylimidazo-
Or diethyl cyanophosphate, diphenylphosphoryl azide, 2,2'-dipyridyl disulfide, or methylpyridinium halide), or treating (IV) with the above-mentioned condensing agent to give azlactone or a base. (Triethylamine, pyridine or DB
U) in the presence of alkoxycarbonyl chloride (ethoxycarbonyl chloride, isobutyloxycarbonyl chloride or benzyloxycarbonyl chloride)
By reacting (V) with (V) as a mixed acid anhydride of (IV).

Solvents used in this reaction are reaction (II) and (III)
The solvent used for can be used. The reaction temperature is −
It is 20 ° C to 150 ° C.

The desired compound (I) thus obtained can be isolated by a conventional separation / purification means such as washing, extraction, concentration, recrystallization and column chromatography.

The compound represented by the formula (I) has a plurality of stereoisomers when it has an asymmetric carbon atom in the molecule, and any one of these stereoisomers or a mixture thereof is the present invention. Included in.

On the other hand, according to the present synthesis method, optical isomers can be separately synthesized if necessary. That is, in that case, the corresponding optical isomer of the formula (I) can be obtained by carrying out the above reaction using a raw material compound that has been optically resolved in advance.

In the condensation reaction of (III) and (V) of Method B, the amino group of (III) is usually protected (the protecting group is isobutyloxycarbonyl or benzyloxycarbonyl). The condensation reaction of (III) and (V) is carried out under the same reaction conditions as the condensation reaction of (IV) and (V) described in Method A. The compound (VI) obtained by the reaction of (III) and (V) can be purified by a conventional method. Got here (V
After removing the amino-group-protecting group of I), the obtained compound is reacted with the compound (II). This reaction is carried out in the same manner as the reaction of (II) and (III) described in Method A to obtain the compound having the general formula (I). Table 1 lists specific compounds having the general formula (I) obtained by the present invention.

[0037]

[Table 1]

[0038]

[Chemical 13]

─────────────────────────────── Compound No. R ¹ X Z R ³ R ⁴ R ⁵ R ⁶ ── ───────────────────────────── 1 Ph CO CH ₂ H CH ₃ CH ₃ Ph 2 Ph CO SH CH ₃ CH ₃ Ph 3 Ph CO CH ₂ HH CH ₃ Ph 4 Ph CO SHH CH ₃ Ph 5 Bz SO ₂ SHH CH ₃ Ph 6 DiFPh CO SHH CH ₃ Ph ───────────────────── ─────────── In the above table, Ph represents a phenyl group, Bz represents a benzyl group, and DiFPh represents a 2,6-difluorophenyl group.

In the table, when the formula (I) has an incorrect carbon, it means that it is one of stereoisomers or a mixture thereof.

The compound represented by the general formula (I) is a novel compound and has an excellent immunostimulating action and antitumor action, and is useful as a therapeutic or preventive agent for various infectious diseases and malignant tumors. Next, its pharmacological activity will be described with reference to typical examples.

1) Immune recovery effect of tumor-bearing mice Erlich-cancer cell transplanted ICR / JCL (female) mice were used as tumor-bearing mice, and non-tumor-bearing mice were used as controls
The delayed allergic reaction was examined as a mouse footpad reaction using BCG as an antigen.

That is, 9 days before immunization, 2 × 10 ⁶ Ehrlich's cancer cells were subcutaneously transplanted into the axilla of a mouse, and the mice were placed in a tumor-bearing state. After administration, immunization, 14 days after immunization, BCG was re-inoculated into the hind footpads, and swelling of the footpads was observed 24 hours later.
G Measured by difference from non-reinoculated footpad. Normal animal controls were similarly immunized with BCG.

The test compound was suspended or dissolved in a physiological saline containing 0.25% CMC, and the suspension was dissolved in 2 days 4 days and 2 days before immunization.
A single dose of 1 mg or 10 mg / kg was orally or intraperitoneally administered.

For comparison, the previously clinically applied antitumor immunotherapeutic agents bestatin and levamisole were administered in the same manner as the test compound.

The recovery of immune function of tumor-bearing animals was calculated by calculating the recovery rate according to the following formula.

Recovery rate = 100-[[(Paw swelling of normal control animal-footpad swelling of test compound-administered animal) / (footpad swelling of normal control animal-footpad swelling of cancer-bearing control animal)] × 10
0] Regarding the immunorestorative effect of the test compound, when the recovery rate is 80% or more at any of the doses, the footpad reaction is the footpad reaction of the normal animal control, based on the results of statistical studies on the control of normal animals. No significant difference was found between the reaction and the response, so it was judged that there was a significant immune recovery effect (+), and the recovery rate was 70.
The case of -80% was judged to have an immune recovery tendency (±). The test results are shown in Table 2.

[0048]

[Table 2] The compound numbers in the table are the compound numbers in Table 4.

From these results, it is recognized that the test compound has an action of restoring the immune response of cancer-bearing mice, which reduces the immune response, to the same level as in normal animals.

Effect on Transplanted Tumor A mouse transplantable experimental tumor, which is a syngeneic tumor, frequently invades or metastasizes into normal tissue from the site of transplantation,
MH-134 mouse liver cancer with high malignancy was selected, and the antitumor effect of the combination with an antitumor chemotherapeutic agent was examined.

That is, 2 × 1 syngeneic MH-134 tumor cells were subcutaneously subcutaneously in the axilla of a C ₃ H / HeN-Crj (female) mouse.
0 ⁶ cells were transplanted, and on the 7th day after the transplantation when the tumor diameter reached about 10 mm, 2 mg / kg of the antitumor chemotherapeutic agent Carbocon was intravenously administered, and at the same time, 0.1 mg / kg and 1.0 mg / kg of the test compound were administered. 1
0 mg / kg was orally administered from the 7th day of tumor implantation for 5 consecutive days.

The test compound is a trace amount of the surfactant Twee.
After dissolving in n 80, physiological saline was added and the solution was administered as a uniform emulsion.

For comparison, the already clinically applied antitumor immunotherapeutic agent bestatin was similarly administered.

The experimental period was up to 60 days after tumor cell transplantation, the average survival time until tumor death was calculated, and the antitumor chemotherapeutic agent alone administration group and the antitumor chemotherapeutic agent were compared with the average survival time of the untreated control group. The average survival time increase rate (life extension rate) of the test compound combination group was calculated. Furthermore, within the experimental period, the response rate was determined from the number of animals in which solid tumors were completely or completely disappeared from the transplanted tumor site.

The antitumor effect of the test compound is the survival rate of the group administered with the antitumor chemotherapeutic agent alone and an increase in the survival rate of the combination group of the antitumor chemotherapeutic agent and the test compound with respect to the response rate (increased survival rate). Based on an increase in response rate (increased response rate) as an index, which is adopted by many tumor research institutes as a criterion for determining the efficacy of antitumor chemotherapeutic agents, the survival rate is 25% or more In either of the above cases, either the case of showing an increased survival rate of 25% or more, or the case of showing an increased response rate of 20% or more from the point that the response rate of the antitumor chemotherapeutic agent single administration group is 0% is satisfied. The case was judged as (+). The test results are shown in Table 3.

[0056]

[Table 3] The compound numbers in the table are the compound numbers in Table 4.

From these results, it is confirmed that the test compound has an excellent antitumor effect when used in combination with an antitumor chemotherapeutic agent.

As is clear from the results of the pharmacological tests in Tables 2 and 3, the amino acid derivative of the present invention acts on the immune system,
It was found that the acquired immunodeficiency state was improved, and that it was excellent as a therapeutic agent for malignant tumors when used in combination with a chemotherapeutic agent or the like or alone.

The amino acid derivative of the present invention is: It is clear that it is excellent as an immunostimulant and a host-mediated anticancer agent for controlling tumors, bacterial infections or autoimmune diseases. The dosage forms include subcutaneous injection, intravenous injection,
Examples include parenteral administration methods such as intramuscular injection and suppositories, and oral administration methods such as tablets, capsules, powders and granules. The dose for an adult varies depending on the disease to be treated, the route of administration and the number of administrations, but usually 5 to 300 mg is administered once or in several divided doses per day.

The amino acid derivative of the present invention can be prepared for administration by any conventional method by analogy with other immunostimulants and anticancer agents. Therefore, the present invention also includes a pharmaceutical composition containing an amino acid derivative suitable as a pharmaceutical for human body. Such compositions are used in conventional manner with any desired pharmaceutical carrier. When these compositions consist of unit doses, each unit contains from 5 to 5
It preferably contains 0 mg of active ingredient.

Next, the present invention will be explained with reference to Reference Examples and Examples, but the scope of the present invention is not limited to these Examples.

[0062]

[Reference example]

Reference example 1. N- (3-trifluoromethylbenzoyl) aminoacetic acid Glycine (2.0 g) and sodium hydroxide (2.1 g) in water (15 ml) with vigorous stirring under ice cooling, 3-trifluoromethylbenzoyl chloride (5.5 g). ) Is slowly added dropwise, and after completion of the addition, the mixture is heated with stirring at 70 ° C. for 2 hours. After allowing to cool, the reaction solution was washed with ethyl acetate and then the aqueous layer was washed with 8
Neutralize with N hydrochloric acid, collect the precipitated crystals by filtration, and dry 4.
6 g of the desired product was obtained.

Similarly, a raw material intermediate represented by the formula (IV) could be synthesized.

Reference Example 2. N-α, α-dimethylbenzyl-3- (benzyloxy
Carbonylamino) propionamide N-benzyloxycarbonyl-β-alanine (6.6 g)
And α, α-dimethylbenzylamine (4.0g) and triethylamine (7.4g) were dissolved in dry methylene chloride (400ml) and 2-chloro-1-methylpyridinium iodide (8.8g) was added slowly at room temperature. After the addition, the mixture is heated under reflux for another 2 hours. The reaction solution is a 10% sodium hydroxide solution,
It is washed with 1N-hydrochloric acid water and then with brine, and anhydrous sodium sulfate is added and dried. Crude crystals (10.7 g) obtained by concentrating the reaction solution under reduced pressure were recrystallized from n-hexane-benzene to obtain 5.6 g of the desired product having a melting point of 96 to 97 ° C.
(Yield 56%) Reference Example 3. N-α, α-dimethylbenzyl-3-aminopropion
Amide Reference Example 2. Of the compound (4.51 g) obtained by the reaction of 1. with palladium-carbon (1.5 g) in methanol-acetic acid-water (8:
2: 1) mixed gas (110 ml) with hydrogen gas at room temperature and atmospheric pressure.
Introduce time. Then, the insoluble matter is filtered off, adjusted to pH 10 with 10% sodium hydroxide solution, and extracted with ethyl acetate (150 ml). The extract was washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the target compound (2.3 g) as an oily matter. Similarly, the starting material intermediate represented by the formula (VI) could be synthesized.

Reference Example 4. N-α, α-dimethylbenzyl-3-benzyloxy-
2- (tert-butoxycarbonylamino) propiona
Amido α, α-dimethylbenzylamine (5.0g) and triphenylphosphine (10.0g) in methylene chloride (200ml) solution 10
N-tert-butoxycarbonyl-O-benzyl (L) serine (11.0 g) was added as a powder at -15 ° C, then 2,
2'-pyridyl disulfide (8.2g) in methylene chloride (20m
l) The solution was added dropwise. After stirring at room temperature for 1 hour, the solvent was evaporated and the residue was subjected to silica gel column chromatography. The target compound was obtained as an oily substance from the elution with ethyl acetate-hexane (2: 1). Yield 9.2g Reference example 5. N-α, α-dimethylbenzyl-3-benzyloxy-
2-Aminopropionamide trifluoroacetic acid salt Anisole (2.4 g) of the compound (9.0 g) obtained in Reference Example 4
− Add trifluoroacetic acid (15.0 ml) to a solution of methylene chloride (100 ml).
g) was added and the mixture was stirred at room temperature for 1 hour. After completion of the reaction, trifluoroacetic acid was distilled off, and ether was added to the residue to obtain 8.8 g of the target compound as needle crystals. Melting point 148-150 ° C.

Reference Example 6. N-α, α-dimethylbenzyl-2- (tert-butoxy
Carbonylamino) -3- (p-methoxybenzylthio)
E ) Propionamide α, α-dimethylbenzylamine (4.95 g) and triphenylphosphine (9.69 g) in methylene chloride solution (150
ml) to N-tert-butoxycarbonyl-s- (p-methoxybenzyl) cysteine ((α) D = −2.3 ° (C =
1, CHCl ₃ ), 12.51 g) and then 2,2′-dipyridine disulfide (8.57 g) were sequentially added, and the mixture was heated under reflux for 2 hours and 15 minutes under a nitrogen stream. After completion of the reaction, methylene chloride was added and the methylene chloride layer was washed with cold dilute hydrochloric acid water, then quickly washed with 3% caustic soda water (cooled with ice), and then washed with cold dilute hydrochloric acid water and brine. The methylene chloride layer is dried with anhydrous sodium sulfate and the solvent is distilled off. The resulting residue (about 30
g) was purified by column chromatography using silica gel (500 g). 15-20 g of hexane containing 15 to 20% ethyl acetate
A crystalline residue of was obtained. Recrystallization from hexane containing ethyl acetate gave 13.3 g of the desired compound. Melting point 81
℃, (α) _D ²⁵ = +6.7 ° (C = 1, CHCl ₃ ) Infrared absorption spectrum ν _max (CHCl ₃ ) cm ^-1 ; 1585,1610,16
80,3430 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm; 1.47 (9H, s), 1.69
(6H, s), 2.75 (2H, dd, J = 5.8Hz, 9Hz), 3.71 (2H, s), 3.78 (3
H, s), 4.15 (1H, t, J = 5.8Hz), 6.8 to 7.5 (9H, m).

Reference Example 7. N-α, α-dimethylbenzyl-2-amino-3- (p
-Methoxybenzylthio) propionamide triflu
Oro acetate N-alpha, alpha-dimethylbenzyl -2- (tert- butoxycarbonylamino)-3-(p-methoxybenzylthio) in chloroform propionamide (6.093g) (20ml)
Thiophenol (1.6 g) was added to the solution, and trifluoroacetic acid (15 ml) was added under ice cooling and the mixture was stirred for 20 minutes. After stirring at room temperature for 1.5 hours, the solvent and reagents were distilled off under reduced pressure and the residue obtained was dissolved in ether (7 ml) and dissolved in hexane (50 ml).
ml) was added to obtain the target compound.

Reference Example 8. N-α, α-dimethylbenzyl-2- (tert-butoxy
Carbonylamino) -6-benzyloxycarbonyla
The amino hexanoate amide N-tert-butoxycarbonyl -N- dicyclohexylamine salt benzyloxycarbonyl-lysine (517 mg) was dissolved in a small amount of water and ethyl acetate, and washed twice with 10% citric acid aqueous solution. The ethyl acetate layer was washed with water and saturated brine and dried over anhydrous magnesium sulfate. After distilling off the solvent, the residue was dissolved in methylene chloride (9 ml), and a solution of dicyclohexylcarbodiimide (209 mg) and α, α-dimethylbenzylamine (145 mg) in methylene chloride (6 ml) was added to this solution, which was stirred at room temperature for 2. Stir for 5 hours. After filtering the precipitate, the filtrate was concentrated under reduced pressure, the residue was dissolved in ethyl acetate, and washed successively with 10% citric acid, 5% aqueous sodium hydrogen carbonate and saturated brine. After drying over anhydrous magnesium sulfate, the solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography with ethyl acetate-hexane (1: 1) to obtain 297 mg of the target compound as an amorphous substance. Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.05-1.86 (7H, m),
1.43 (9H, s), 1.62,1.64 (6H, s x 2), 3.11 (2H, br.d., J = 6H
z), 3.72-4.28 (1H, br.), 4.68-5.29 (3H, m), 6.40-6.65 (1H,
br.), 6.97-7.38 (10H, m).

Reference Example 9. N-α, α-dimethylbenzyl-2-amino-6- (beta
Benzyloxycarbonylamino) hexanoamide N-α, α-dimethylbenzyl-2- (tert-butoxycarbonylamino-6-benzyloxycarbonylaminohexanoamide (2.18 g) with anisole (0.47 g) and trifluoroacetic acid (10 ml) was added and the mixture was stirred at room temperature for 30 minutes.
After completion of the reaction, trifluoroacetic acid was distilled off under reduced pressure, and the residue was mixed with 5
% Sodium hydrogen carbonate solution is added to neutralize and the mixture is extracted with ethyl acetate. The organic layer is extracted with 10% aqueous citric acid solution, the aqueous layer is neutralized with 5% aqueous sodium hydrogen carbonate and then extracted again with ethyl acetate. The extract was dried over anhydrous magnesium sulfate and the solvent was distilled off under reduced pressure to obtain 1.01 g of the objective compound as an amorphous substance.

Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.10
-2.08 (7H, m), 1.61,1.64 (6H, s × 2), 3.11 (2H, br.d., J = 5.8
Hz), 4.26-4.74 (1H, br.), 4.97-5.34 (3H, m), 6.93-7.15 (1
H, br.s), 7.15-7.56 (10H, m), 7.77-8.12 (1H, m).

Reference Example 10. N-α, α-dimethylbenzyl-2-benzoylamino
Acetamide Dry methylene chloride containing α, α-dimethylbenzylamine (1.0 g) and triphenylphosphine (1.95 g)
Hippuric acid (1.33 g) and 2,2'-dipyridyl disulfide (1.63 g) were sequentially added to (100 ml), and the mixture was heated under reflux for 2 hours. After completion of the reaction, the reaction solution is washed with a 10% sodium hydroxide aqueous solution, then with a 1N hydrochloric acid aqueous solution, and then dried over anhydrous sodium sulfate. The reaction mixture was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography for separation and purification (hexane-ethyl acetate mixed solvent) to give 1.97 g of the desired product. Yield 90%, melting point: 156-157 ° C Elemental analysis (C ₁₈ H ₂₀ N ₂ O ₂ ) Calculated value: C, 72.95; H, 6.80; N, 9.45 Analytical value: C, 72.99; H, 6.89; N, 9.45 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3380,3280,
1680,1645 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.72 (6H, s), 4.21
(2H, d, J = 5Hz), 7.2-7.7 (10H, m), 7.8-8.0 (2H, m).

Reference Example 11. N- (1,2,2-trimethylpropyl) -2-benzo
Ilaminoacetamide 3-amino-2,2-dimethylbutane (1.0 g) and hippuric acid
Dicyclohexylcarbodiimide (2.2 g) was added to an ethyl acetate solution (30 ml) of (1.7 g) under ice-cooling, the mixture was stirred at room temperature for 2 hours, and left to stand overnight. After the precipitate is filtered off, the filtrate is washed with 10% sodium hydroxide solution, then with 1N hydrochloric acid, and then with brine, and the reaction filtrate is dried over anhydrous sodium sulfate. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 1.65 g of the desired product. Yield 64%, melting point: 124-126 ° C Elemental analysis (C ₁₅ H ₂₂ N ₂ O ₂ ) Calculated value: C, 68.67; H, 8.45; N, 10.68 Analytical value: C, 68.65; H, 8.60; N, 10.65 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3300,1680,
1640 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 0.90 (9H, s), 1.09
(3H, d, J = 7Hz), 3.80 (1H, qd, J = 7Hz, 3Hz), 4.18 (2H, d, J = 7H
z), 6.9-7.1 (1H, br.s), 7.3-7.55 (3H, m), 7.65-8.0 (3H,
m).

Reference Example 12 N-α-methylbenzyl-2- (2,6-dichloroben
Zoylamino) acetamide In a nitrogen stream, a solution of N- (2,6-dichlorobenzoyl) glycine (1.74g) and triethylamine (0.71g) in dry tetrahydrofuran (30ml) was slowly added to isobutyl chloroformate at -20 ° C. (1.05 g) is added dropwise, and the mixture is stirred for 20 minutes. Then, α-phenethylamine (0.84 g) was added, the mixture was stirred at -10 to -20 ° C for 30 minutes, and the reaction solution was extracted with ethyl acetate (60 ml). After washing the extract with 10% sodium hydroxide solution and 1N hydrochloric acid,
Further, it is washed with brine and dried over anhydrous sodium sulfate. The reaction solution was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to obtain 1.58 g of the desired product. Yield 67%, melting point 155 ° C. Elemental analysis (C ₁₇ H ₁₆ N ₂ O ₂ Cl ₂ ) Calculated value: C, 58.13; H, 4.59; N, 7.98; Cl, 20.19 Analysis value: C, 57.98; H, 4.68 ; N, 8.02; Cl, 20.34 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3340,3230,
1645 Nuclear magnetic resonance spectrum (DMSO-d ₆ ) δppm: 1.40 (3H, d, J = 7H
z), 3.87 (2H, d, J = 6Hz), 4.90 (1H, qd, J = 7Hz, 6Hz), 7.07-8.
87 (10H, m).

Reference Example 13. N-α, α-dimethylbenzyl-3- (benzoylami)
B) Benzoyl chloride (0.3 g) was slowly added dropwise to a toluene solution (10 ml) of N-α, α-dimethylbenzyl-3-aminopropionamide (0.4 g) and triethylamine (0.22 g), and the mixture was further added for 2 hours. Stir. Then ethyl ether (20m
After diluting with l), wash with water and dry over anhydrous sodium sulfate. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography to obtain 0.52 g of the desired product.

Yield 87%, melting point 147-148 ° C. Elemental analysis (C ₁₉ H ₂₂ N ₂ O ₂ ): Calculated value; C, 73.52; H, 7.14; N,
9.02 analytical value; C, 72.92; H, 7.15; N,
9.02 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3300,1640 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.69 (6H, s), 2.5
4 (2H, t, J = 6Hz), 3.71 (2H, td, J = 6Hz, 6Hz), 7.2-7.6 (10H,
m), 7.7-8.0 (2H, m).

Reference Example 14. N-α, α-dimethylbenzyl-2-benzoylamino
-3-Benzyloxypropionamide An oil (3.2 g) obtained by treating the compound obtained in Reference Example 5 with triethylamine and triethylamine (1.5 g) were dissolved in methylene chloride (150 ml), and ice-cooled to this solution. Benzoyl chloride (1.5 g) was added dropwise under cooling. After stirring the mixed solution for 30 minutes, the methylene chloride layer was washed with water and dried over anhydrous sodium sulfate. The solvent was distilled off and the residue was recrystallized from ether-hexane to obtain 4.1 g of the objective compound. Melting point 88-90
Elemental analysis (C ₂₆ H ₂₈ N ₂ O ₃ ): Calculated value; C, 74.97; H, 6.78; N, 6.73 Analytical value; C, 74.74; H, 6.85; N, 6.68 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3250,166
0,1630 Reference example 15. N-α, α-dimethylbenzyl-3-hydroxy-2-
10% of (benzoylamino) propionamide N-α, α-dimethylbenzyl-2-benzoylamino-3-benzyloxypropionamide (2.0 g)
Acetic acid (4 ml) -ethanol (60 m) in the presence of Pd / C (0.4 g)
catalytic reduction in l). After completion of the reaction, the catalyst was filtered off and the solvent was distilled off. The obtained residue was recrystallized from acetic acid-hexane to obtain 1.4 g of the target compound.

Melting point 152-153 ° C. Elemental analysis (C ₁₉ H ₂₂ N ₂ O ₃ ): Calculated value; C, 69.92; H, 6.79; N, 8.58 Analytical value; C, 70.03; H, 6.87; N,
8.66 Infrared absorption spectrum ν _max (nujol) cm
⁻¹ : 3400, 3350, 1660, 1630 Reference Example 16. N-α, α-dimethylbenzyl-2-benzoylamino
-3- (p-Methoxybenzylthio) propionamide The compound obtained in Reference Example 6 was dissolved in toluene (50 ml), triethylamine (4.70 ml) was added, and benzoyl chloride (2.23 g) was gradually added under ice cooling. . After stirring for 70 minutes, ethyl acetate was added to the reaction solution, washed with brine, and dried over anhydrous sodium sulfate. The solvent is distilled off and the obtained residue is purified by column chromatography using silica gel (210 g). From the hexane outlet containing 30-40% ethyl acetate, 4.31 g of an oily substance was obtained. Ethyl acetate-
Crystals that dissolve and precipitate in a hexane mixture (about 250 mg, (α) _D
(0, mp 189 ° C.) and the mother liquor was concentrated to give 3.61 g of the glassy target compound. (Α) _D ²⁵ = −25.3
° (C = 1, CHCl ₃ ) infrared absorption spectrum ν _max (CHCl ₃ ) cm ^-1 : 1581,161
1,1650,1680,3330,3420 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.68 (6H, s), 2.6
-3.2 (2H, m) 3.77 (2H, s), 3.81 (3H, s), 4.75 (1H, m), 6.7-7.9
(16H, m) MS m / e: 462 (M ⁺ ), C ₂₇ H ₃₀ O ₃ N ₂ S Reference example 17. N-α, α-dimethylbenzyl-2-benzoylamino
-3-Mercaptopropionamide N-α, α-dimethylbenzyl-2-benzoylamino-3- (p-methoxybenzylthio) propionamide
(1.010 g) was dissolved in trifluoroacetic acid (10 ml), anisole (1.0 ml) was added, and Hg (OAc) ₂ (793
mg) was added and stirred. After stirring for 1.5 hours, the solvent was distilled off at room temperature, ether was added to the residue, and the precipitate was collected by filtration. The obtained precipitate (mercury salt) was dissolved in DMF (50 ml) and hydrogen sulfide gas was passed under ice cooling. After 1.5 hours, the precipitated HgS was removed by filtration through Celite, and the filtrate was concentrated under reduced pressure at room temperature to obtain 702 mg of the desired product as crystals. Melting point 169-173 ℃ Infrared absorption spectrum ν _max (CHCl ₃ ) cm ^-1 : 1585,160
7,1650,1680,3330,3430 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 1.68 (6H, s), 2.7
-3.3 (2H, m) 4.90 ( 1H, m), 7.1-8.0 (10H, m) MS m / e: 342 (M +), 343 (M + +1), C 19 H 22 O 2
N ₂ S Reference Example 18. N-α, α-dimethylbenzyl-2-benzoylamino
-6-Benzyloxycarbonylaminohexanoamide N-α, α-dimethylbenzyl-2-amino-6-benzyloxycarbonylaminohexanoamide (2.14 g) and pyridine (0.51 g) in methylene chloride (80 ml) solution. Benzoyl chloride (0.76g) in methylene chloride (30ml) with stirring under ice cooling
The solution is added dropwise over 10 minutes. The mixture is stirred at room temperature for 1.5 hours, the solvent is evaporated under reduced pressure, and ethyl acetate and a small amount of water are added to the residue to dissolve it. After adding a 10% aqueous citric acid solution, the mixture is extracted with ethyl acetate. The extract was washed with 5% aqueous sodium hydrogen carbonate and saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. When the residue was subjected to silica gel column chromatography, 2.06 g of the desired compound was obtained in an amorphous form from the portion which was eluted with ethyl acetate-hexane (1: 1). Mp 53 to 58 ° C. (decomposition) Elemental analysis _{(C 30 H 35 N 3 O} 4): Calculated; C, 71.83; H, 7.03 ; N, 8.38 Analytical values; C, 71.28; H, 6.83 ; N, 8.29 IR Absorption spectrum ν _max (nujol) cm ^-1 : 3280,172
0,1700,1655,1635 Nuclear magnetic resonance spectrum (CDCl ₃ ) δppm: 0.95-2.10 (6H,
m), 1.60 (6H, br.s), 3.12 (2H, br.d, J = 5.8Hz), 4.53-5.30 (4
H, m), 6.91-7.93 (17H, m).

Reference Example 19. N-α, α-dimethylbenzyl-6-amino-2-ben
Zoylaminohexanoamide N-α, α-dimethylbenzyl-2-benzoylamino-6-benzyloxycarbonylhexanoamide (1.73
g) 10% Pd in ethanol (80 ml) -1N hydrochloric acid (4 ml)
/ C (0.33 g), catalytic reduction was performed at room temperature. After the catalyst was filtered off, the filtrate was concentrated under reduced pressure to obtain 1.35 g of the desired compound in an amorphous form. Melting point 94-101 ° C (decomposition) Elemental analysis (C ₂₂ H ₂₉ N ₃ O ₂ · HCl · 1/3 H ₂
O): Calculated value; C, 64.46; H, 7.54; N,
10.25 Analytical value; C, 64.11; H, 7.70; N,
9.92 Infrared absorption spectrum ν _max (nujol) cm ^-1 : 3250,165
5,1635

[0079]

EXAMPLES The compounds listed in Table 4 could be obtained according to the methods of Reference Examples 1 to 19.

[0080]

[Table 4]

[0081]

[Chemical 14]

────────────────────────────── Compound No. R ¹ X Z R ³ R ⁴ R ⁵ R ⁶ Melting point ° C. ───────────────────────────── 1 Ph CO CH ₂ H CH ₃ CH ₃ Ph 119 to 120 2 Ph CO SH CH ₃ CH ₃ Ph Gum Shape 3 Ph CO CH ₂ HH CH ₃ Ph Gum Shape 4 Ph CO SHH CH ₃ Ph Gum Shape ─────────────────────────── —————————————————————————————————————————————————————————————————————————————————————————————————————————————————————————————— from (S) (1) to (+) are in the table above, then (s) are S-coordinates.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical indication C07D 333/38 // A61K 31/165 ADU 8413-4C 31/38 ABD 9360-4C 31/40 9360 −4C 31/425 9360−4C 31/455 9360−4C (72) Inventor Somei Shimizu 1-258 Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd.

Claims (1)

[Claims] 1. A general formula: [Wherein R ¹ is a benzyl group, a phenyl group which may have a substituent (the substituent is a halogen atom (fluorine, chlorine, bromine).), And 1 to 3 of these substituents are included. X) represents a —CO— group or a —SO ₂ — group, Z represents a methylene group or a sulfur atom, R ³ represents a hydrogen atom, and R ⁴ represents a hydrogen atom or a carbon number of 1 to 1. 4 represents an alkyl group, R ⁵ represents an alkyl group having 1 to 4 carbon atoms, and R ⁶ represents a phenyl group. ] The compound which has.