JPS6334146B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the general formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
A mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and A is an alkyl group that may have a hydroxyl group, an alkoxy group, an amino group, a halogen atom, or a carbamoyl group. It represents a substituted amino group, hydrazino group, lower alkyl group or aryl substituted hydrazino group, hydroxyamino group, lower alkoxyamino group or aralkyloxyamino group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a group other than a hydrogen atom, a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. represents the above group. )
The present invention relates to an amino acid derivative represented by the above formula, a pharmacologically acceptable salt thereof, a method for producing the same, and an immunostimulant or anticancer agent containing the present compound as an active ingredient. In the general formula (), R ₁ is preferably a hydrogen atom; for example, methyl, ethyl, n-propyl,
Isopropyl, n-butyl, isobutyl, tert-
Straight chain or branched alkyl group having 1 to 4 carbon atoms such as butyl; halogenoalkyl having 1 to 2 carbon atoms such as trifluoromethyl, 2,2,2-trichloroethyl groups; e.g. halogen atoms such as chlorine, bromine, iodine, fluorine; mercapto groups; e.g. methylthio,
A linear or branched alkylmercapto group having 1 to 16 carbon atoms such as ethylthio, n-propylthio, isopropylthio, n-butylthio, tert-butylthio, and hexadecylthio;
Hydroxyl group; for example, a linear or branched alkoxy group having 1 to 16 carbon atoms such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, hexadecyloxy; for example, acetoxy , propanoyloxy, n-butyryloxy, isobutyryloxy,
A linear or branched alkanoyloxy group having 2 to 16 carbon atoms such as pivaloyloxy and palmitoyloxy, or a lower alkyl group such as methyl, ethyl, n-propyl, and isopropyl as a substituent on an aromatic ring, lower alkoxy groups such as methoxy, ethoxy, n-propoxy, and isopropoxy, halogen atoms such as chlorine, bromine, iodine, and fluorine, nitro groups, trifluoromethyl groups, cyano groups, hydroxyl groups, amino groups, or carboxy groups. Acyloxy groups, such as benzoyloxy groups with or without; carboxy groups; e.g. methoxycarbonyl, ethoxycarbonyl, n-
A linear or branched alkoxycarbonyl group having 2 to 17 carbon atoms such as propoxycarbonyl, isopropoxycarbonyl, n-butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, and hexadecyloxycarbonyl; an amino group; For example, lower alkyl-substituted amino groups such as methylamino, ethylamino, n-propylamino, isobutylamino, dimethylamino, diethylamino, and diisopropylamino; Alkyl groups, lower alkoxy groups such as methoxy, ethoxy, n-propoxy, and isopropoxy, halogen atoms such as chlorine, bromine, iodine, and fluorine, nitro groups, trifluoromethyl groups, hydroxyl groups, amino groups, or carboxy groups. Aryl-substituted amino groups, such as phenylamino, with or without; linear or branched carbon atoms 2 to 16, such as acetylamino, propionylamino, n-butyrylamino, isobutyrylamino, pivaloylamino, palmitoylamino; A lower alkyl group such as methyl, ethyl, n-propyl, isopropyl, a lower alkoxy group such as methoxy, ethoxy, n-propoxy, isopropoxy as a substituent on an alkanoylamino group or an aromatic ring,
Halogen atoms such as chlorine, bromine, iodine, and fluorine, nitro groups, trifluoromethyl groups, hydroxyl groups,
acylamino groups, such as benzoylamino groups with or without amino or carboxy groups;
For example, halogeno lower alkoxycarbonylamino groups such as 2,2,2-trichloroethoxycarbonylamino, 2,2,2-tribromoethoxycarbonylamino; carbon numbers such as methanesulfonyl, ethanesulfonyl, propanesulfonyl, butanesulfonyl Alkanesulfonyl group having 1 to 4 atoms; represents a nitro group or a cyano group, and R ₂ and R ₃ are the same or different and are hydrogen atoms or a linear or branched group such as methyl, ethyl, n-propyl, isopropyl. Represents a chain alkyl group having 1 to 3 carbon atoms, R ₄
is a hydrogen atom; for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,
Straight chain or branched alkyl group having 1 to 4 carbon atoms such as tert-butyl; cyano group; cyano lower alkyl group such as cyanomethyl, 2-cyanoethyl, 3-cyanopropyl, 4-cyanobutyl ; Hydroxy lower alkyl groups such as hydroxymethyl, 2-hydroxyethyl, 4-hydroxybutyl; Lower alkoxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, n-butoxycarbonyl, isobutoxycarbonyl; Carboxy group; aromatic Methyl, ethyl, n- as a substituent on the ring
lower alkyl groups such as propyl and isopropyl, lower alkoxy groups such as methoxy, ethoxy, n-propoxy, and isopropoxy, trifluoromethyl groups, cyano groups, hydroxyl groups, nitro groups, amino groups, or fluorine, chlorine, and bromine. Aryl groups such as phenyl with or without halogen atoms; lower alkyl groups such as methyl, ethyl, n-propyl, isopropyl, lower alkyl groups such as methoxy, ethoxy, n-propoxy as substituents on the aromatic ring; Aralkyl groups such as benzyl, phenethyl, phenylpropyl with or without alkoxy, trifluoromethyl, cyano, hydroxyl, nitro, amino or halogen atoms such as fluorine, chlorine, bromine; mercaptomethyl , mercaptoethyl, mercaptopropyl,
a mercapto lower alkyl group such as mercaptobutyl; a lower alkylthio lower alkyl group such as methylthiomethyl, methylthioethyl, methylthiobutyl, propylthioethyl or, for example, when the compound () is of the formula (In the formula, R ₁ , R ₂ , R ₃ , A and n are the same as defined above.) represents a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and A is Hydroxyl group; for example, a linear or branched alkoxy group having 1 to 16 carbon atoms such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, hexyloxy, decyloxy, hexadecyloxy ; Amino group; For example, lower alkyl substituted amino group such as methylamino, dimethylamino, ethylamino, diethylamino, n-propylamino, isopropylamino, n-butylamino, isobutylamino; For example, 2-chloroethylamino, 2-bromo ethylamino, 3-
Halogeno lower alkylamino groups such as chloropropylamino; carbamoyl lower alkylamino groups such as 2-carbamoylethylamino and 3-carbamoylpropylamino; hydrazino groups such as methylhydrazino, N,N-dimethylhydrazino, Lower alkyl-substituted hydrazino groups such as ethylhydrazino and N,N-diethylhydrazino; for example, lower alkyl groups such as methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy as substituents on the aromatic ring , aryl-substituted hydrazino groups such as phenylhydrazino with or without lower alkoxy groups such as isopropoxy or halogen atoms such as fluorine, chlorine, bromine; hydroxyamino groups such as methoxyamino, ethoxyamino, n
- a lower alkoxyamino group such as propoxyamino, isopropoxyamino, or as a substituent on the aromatic ring, such as methyl, ethyl, n-propyl,
aralkyloxy such as benzyloxyamino with or without a lower alkyl group such as isopropyl, a lower alkoxy group such as methoxy, ethoxy, n-propoxy, isopropoxy or a halogen atom such as fluorine, chlorine, bromine; Indicates an amino group. In the general formula (), the carboxylic acid or hydroxamic acid compound in which the substituent A is a hydroxyl group or a hydroxyamino group can be in the form of a pharmacologically acceptable salt, if necessary. Such salts include lithium, sodium, potassium,
Examples include salts of inorganic metals such as calcium, magnesium and aluminum, and ammonium salts such as ammonium, cyclohexylammonium, diisopropylammonium and triethylammonium, preferably sodium salts, potassium salts and calcium salts. In addition, in the compound having the general formula (), optical isomers based on asymmetric carbon atoms exist,
Although all of these isomers are represented by a single formula, this is not intended to limit the scope of the present description. If necessary, optical isomers can be obtained by conventional methods, such as optical resolution of the target compound (2) or stereospecific synthesis using an optically active compound as a reaction raw material. The amino acid derivative having the general formula () obtained by the present invention is a novel compound, and is a compound useful as a medicine that exhibits excellent immunostimulatory and antitumor effects. The present inventors have repeatedly studied the synthesis and pharmacological activity of amino acid derivatives with the aim of developing host-mediated anticancer agents. As a result, the novel amino acid derivative represented by the above general formula () has immunostimulatory and antitumor effects. The present invention was completed by discovering that the compound is useful as a medicine. The novel compound according to the present invention can be produced by the method shown below. Method 1 Formula (In the formula, R ₂ , R ₃ and R ₄ have the same meanings as defined above, and M ₁ represents a linear or branched alkyl group having 1 to 16 carbon atoms.) the expression (In the formula, R ₁ and n have the same meanings as described above, and X represents a halogen atom such as chlorine or bromine.) The expression (wherein R ₁ , R ₂ , R ₃ , R ₄ , M ₁ and n have the same meanings as described above) can be obtained. The reaction proceeds equimolar stoichiometrically. This reaction is carried out in the presence of a base, and the bases used include organic bases such as tertiary amines such as triethylamine, dimethylaniline, diethylaniline, and pyridine, or inorganic bases such as alkali metal hydroxides such as potassium hydroxide; Alkali metal bicarbonates such as potassium bicarbonate and sodium bicarbonate, alkaline earth metal hydroxides such as calcium hydroxide and magnesium hydroxide, etc. can be used, but the reaction system in an organic solvent is the above-mentioned tertiary amine. In a reaction system using a mixed solvent with water, it is preferable to use an alkali metal bicarbonate such as sodium bicarbonate or potassium bicarbonate. In addition, the solvent used is not particularly limited, but methanol, ethanol,
n-propanol, isopropyl alcohol, n
-Alcohols such as butanol, benzene,
Aromatic hydrocarbons such as toluene and xylene,
Ketones such as acetone, methyl butyl ketone, methyl amyl ketone, halogenated hydrocarbons such as tetrachloroethane, chlorobenzene, dichlorobenzene, nitriles such as benzonitrile, acetonitrile, N,N-dimethylformamide, N, Amides such as N-dimethylacetamide and hexamethylphosphoric triamide, dimethyl sulfoxide, mixed solvents of these solvents, and mixed solvents of these solvents and water are used. There is no particular limitation on the reaction temperature, but -20℃ to
It is in the range of 100°C, preferably 0°C to 30°C. The reaction time varies depending on the type of raw material compound and the reaction temperature, but is usually 1 to 5 hours. After completion of the reaction, the target compound () is collected from the reaction mixture according to a conventional method. For example, if the target compound precipitates from the reaction system, it can be removed, or if it does not precipitate, it can be obtained as a residue by distilling off the solvent. Further, if necessary, it can be further purified by recrystallization, vacuum distillation, chromatography, etc. Amino acids (), which are the raw materials used to carry out this reaction, are Tetrahedron Letter No. 17, 1455-
1458 pages (1977) (Tetrahedron Letters No.17,
P. 1455-1458 (1977)) as follows. That is, the expression (In the formula, R ₄ and M ₁ have the same meanings as described above.) A strong base such as trimethylbenzylammonium hydroxide [(40% methanol solution) (Triton B)], 1, 5
-Diazabicyclo[4,3,0]-5-nonene (DBN), 1,8-diazabicyclo[5,4,0]
Formula using −7-undecene (DBU) etc. (In the formula, R ₂ and R ₃ have the same meanings as described above.) A formula obtained by reacting a compound having the following: (wherein R ₂ , R ₃ , R ₄ and M ₁ have the same meanings as described above) is hydrolyzed with an acid such as a mineral acid such as hydrochloric acid, and neutralized with a base. obtained by. In addition, the formula obtained by hydrolysis (In the formula, R ₂ , R ₃ , R ₄ and M ₁ have the same meanings as above, and X represents a halogen atom or a sulfuric acid group.)
It is also possible to use salts of amino acids having the following as intermediates in the reaction without isolation in a mixed solvent with water. Next, by hydrolyzing the compound () thus obtained with an alkali metal hydroxide in an aqueous solution or a mixed solvent with water, the formula (In the formula, M ₃ represents a metal such as sodium or potassium, and R ₁ , R ₂ , R ₃ , R ₄ and n have the same meanings as described above.) It will be done. The reaction proceeds stoichiometrically at equimolar, but
Preferably, 1.0 to 2.0 mol of caustic alkali is used. The mixed solvent used with water is not particularly limited as long as it is inert to this reaction, but examples include methanol, ethanol, n-propanol,
Alcohols such as isopropyl alcohol and n-butanol, ketones such as acetone and methyl butyl ketone, and the like are used. Further, the caustic alkali used is an alkali hydroxide such as sodium hydroxide or potassium hydroxide. The reaction temperature is preferably 0°C to 50°C. After the reaction is completed, the target product () is collected from the reaction mixture according to a conventional method. For example, if the target product precipitates from the reaction solution, it may be washed with an insoluble solvent, or if it precipitates, it may be added to the reaction solution using an organic solvent such as alcohols such as methanol or ethanol, or ketones such as acetone or methyl ethyl ketone. By adding the following, the desired product () can be precipitated. Next, the compound () obtained in this way is neutralized with an acid in an aqueous solution or a mixed solvent of water and an organic solvent according to a conventional method, so that the formula A carboxylic acid having the formula (wherein R ₁ , R ₂ , R ₃ , R ₄ and n have the same meanings as described above) is obtained. After the reaction is completed, the target product () is collected from the reaction mixture according to a conventional method. For example, if the target product precipitates from the reaction solution, it can be obtained by removing it, or if it does not precipitate, it can be obtained by distilling off the mixed organic solvent, extracting with an organic solvent that is difficult to mix with water, and then distilling off the solvent. can. The extraction organic solvent is not particularly limited as long as it is a solvent that is difficult to mix with water, but examples include ethers such as dimethyl ether and diethyl ether, halogenated hydrocarbons such as dichloromethane and chloroform, and aromatics such as benzene, toluene, and xylene. Group hydrocarbons, esters such as ethyl acetate are used. Further, if necessary, it can be further purified by recrystallization, chromatography, etc. Method 2 formula (In the formula, R ₁ , R ₄ , M ₁ and n have the same meanings as above.) (In the formula, R ₂ and R ₃ have the same meanings as described above.) By reacting a compound having the formula in the presence of a strong base, (wherein R ₁ , R ₂ , R ₃ , R ₄ , n and M ₁ have the same meanings as described above) can be obtained. Although the reaction proceeds stoichiometrically, the compound ()
1.0 to 1.5 mol of compound () per 1 mol
Preferably, 1.0 to 1.5 mol of base is used.
The base used in this reaction is 1,5-diazabicyclo[3,4,0]nonene-5 (DBN), 1,
8-Diazabicyclo[5,4,0]undecene-
7 (DBU) or a strong base such as a metal hydride such as sodium hydride, potassium hydride. The solvent used may be any solvent as long as it is inert to the reaction, such as chloroform, dichloromethane, halogenated hydrocarbons such as dichloroethane, dimethyl ether, diethyl ether,
Examples include ethers such as diisopropyl ether, dioxane, and tetrahydrofuran, and amides such as N,N-dimethylformamide, N,N-diethylacetamide, and hexamethylphosphoric triamide. The reaction temperature ranges from 0°C to 50°C, preferably from 20°C to 30°C. The reaction time varies depending on the type of raw material compound and the reaction temperature, but is usually 2 to 20 hours. After completion of the reaction, the desired product (2) can be obtained by washing with water and distilling off the solvent. If necessary, it can be further purified by recrystallization, vacuum distillation, chromatography, etc. Method 3 Formula (In the formula, R ₁ , R ₂ , R ₃ , R ₄ and n have the same meanings as described above.) A compound having the formula HM ₂ () (wherein M ₂ is Each represents an alkyl-substituted amino group, a hydrazino group, a lower alkyl- or aryl-substituted hydrazino group, a hydroxylamino group, a lower alkoxyamino group, or an aralkyloxyamino group which may have a corresponding amino group, halogen atom, or carbamoyl group. ) by reacting with an organic base having the formula (wherein R ₁ , R ₂ , R ₃ , R ₄ , n and M ₂ have the same meanings as described above) can be obtained. Although the reaction proceeds stoichiometrically, it is preferable to use 1.0 to 1.5 moles of amine. There is no particular problem with the solvent used as long as it is inert to the reaction, but examples include hydrocarbons such as benzene, toluene and xylene, ethers such as dioxane, tetrahydrofuran and diisopropyl ether, dichloroethane and chloroform. Examples include halogenated hydrocarbons. Reaction temperature is 0℃
It is carried out at a temperature ranging from 100℃ to 100℃, but usually from 20℃ to 50℃.
℃, and the time required for the reaction is 2 to 12 hours. After completion of the reaction, the residue can be obtained as the desired product (XI) by washing with water and distilling off the solvent. If necessary, it can be purified by recrystallization, chromatography, etc. The oxazolones having the general formula (), which are the raw material compounds for this method, are described in Journal of Practical Science, Vol. 82, p. 60 (1910) (J. Parkt.
Chem. [2], 82 , 60 (1910)). Method 4 Formula (In the formula, R ₁ , R ₂ , R ₃ , and n have the same meanings as described above.) By dissolving the amino acid having the formula in a solvent and heating it, the formula (wherein R ₁ , R ₂ , R ₃ and n have the same meanings as described above) can be obtained. Any solvent can be used as long as it does not affect the reaction, and examples include ethers such as diisopropyl ether, tetrahydrofuran, and dioxane, and hydrocarbons such as benzene, toluene, and xylene.
To facilitate the reaction, an organic base such as triethylamine, dimethylaniline, or pyridine can be used. Furthermore, compound (XII) can be dissolved alone in a tertiary organic base such as pyridine or diethylaniline, and then heated. Even if you twist it, you can still get the object (). The reaction temperature is carried out in the range of 30°C to 150°C, but usually it is 50°C to 120°C.
Reaction time is 1 to 8 hours. After the completion of the reaction, the target product () is collected according to a conventional method, but if a base such as those mentioned above is used in the reaction, the target product can be obtained by washing the reaction solution with water and distilling off the organic solvent after the reaction. You can get things (). If necessary, it can also be purified by recrystallization, vacuum distillation, chromatography, etc. Furthermore, compound () is esterified to form the formula A compound having the formula (wherein R ₁ , R ₂ , R ₃ , M ₁ and n have the same meanings as described above) is obtained. The esterification may be carried out according to a conventional method, but preferably an amide such as N,N-dimethylformamide, N,N-diethylacetamide, or hexamethylphosphoric acid triamide is used as a solvent, and a halogen is used as an alkylating agent. When carbonized hydrocarbons are used, alkali carbonates such as potassium carbonate and sodium carbonate are preferably used as the base. The amino acid derivatives of the present invention having the general formula () exhibit excellent immunostimulatory and antitumor effects, and representative examples will be described below regarding their pharmacological activities. 1 Immune recovery effect in tumor-bearing mice Ehrlich cancer cell-transplanted ICR/Jcl (female) mice were used as tumor-bearing mice, non-tumor-bearing mice were used as controls, and BCG was used as an antigen to induce delayed allergic reactions as mouse footpad reactions. Examined. That is, 9 days before immunization, Ehrlichi cancer cells 2
×10 ⁶ cells were subcutaneously transplanted into mice to make them tumor-bearing, and 500 μg of BCG was subcutaneously administered to 10 tumor-bearing mice per group. 14 days after immunization, BCG was reintroduced into the skin of the hind footpad. BCG the swelling of the footpad 24 hours after vaccination.
Measured by the difference from non-reinoculated footpads. A normal animal control was similarly immunized with BCG. The test results are shown in Table-1.

[Table] From the table, it can be seen that the test compound has the effect of restoring the immune response of tumor-bearing mice with a decreased immune response. 2. Antitumor effect against mouse metastatic tumor (Sarcoma-37) Sarcoma was administered to ICR/Jcl mice (female, 7 weeks old).
-37 cancer cells were transplanted subcutaneously into the axilla, 2 x 10 ⁶ cells,
After 1 to 4 days and 7 to 10 days, the specimen was intraperitoneally administered once a day. Tumor diameter was measured after 21 days. The test results are shown in Table-2.

[Table] As is clear from the above pharmacological test examples, the amino acid derivatives of the present invention are excellent as immunostimulants and host-mediated anticancer agents for controlling tumors, bacterial infections, and autoimmune diseases. it is obvious. The administration form includes parenteral administration using subcutaneous injection, intravenous injection, intramuscular injection, suppository, etc., and oral administration using tablets, capsules, powders, granules, etc. The dosage for adults varies depending on the target disease, administration route, and number of administrations, but the usual dosage is 5 to 300 mg per day.
Administer in one or several doses. The amino acid derivatives of the present invention can be prepared for administration in any conventional manner by analogy with other immunostimulants and anticancer agents. Therefore, the present invention also encompasses pharmaceutical compositions containing amino acid derivatives suitable as human medicines. Such compositions are provided for use in a conventional manner with any required pharmaceutical carrier. When these compositions consist of unit doses, each unit preferably contains 5 to 50 mg of active ingredient. Next, the present invention will be explained in more detail with reference to Examples and Formulation Examples. Example 1 N-(3,4-dihydroxybenzoyl)-2-
(2'-Cyanoethyl)alanine methyl ester 2-(2'- After adding 1.56 g of cyanoethyl)alanine methyl ester and 1.40 ml of triethylamine and further stirring for 1 hour at 0 to 5°C, the reaction solution was washed with water (30 ml), and the organic layer was dried over magnesium sulfate. The residue obtained by distilling off the solvent was dissolved in 10 ml of methanol, 8.3 ml of aqueous ammonia (28%) was added, and the mixture was further stirred at room temperature for 23 hours, and the reaction solution was heated to 5°C.
Cool, adjust the pH to 2.0 with concentrated hydrochloric acid, and add 50ml of water.
After adding and extracting with dichloromethane (50ml x 2),
Dry the organic layer over magnesium sulfate. The residue obtained by distilling off the solvent was chromatographed on silica gel (developing agent: cyclohexane: ethyl acetate =
When purified with 2:1), a colorless caramel-like target product is obtained.
Obtain 1.60g. Elemental analysis value as C ₁₄ H ₁₆ N ₂ O ₅ Calculated value: C, 57.53; H, 5.52; N, 9.58 Actual value: C, 57.76; H, 5.88; N, 9.09 Illustrated below using the same method. Amino acid derivatives are obtained.

【table】

[Table] Example 2 2-(p-chlorobenzoylamino)-2-
(2'-cyanoethyl)disodium malonate 2
Water salt (a) and 2-(p-chlorobenzoylamino)-2-(2'-cyanoethyl)malonic acid (b) Diethyl N-(p-chlorobenzoyl)-2-(2'-cyanoethyl)aminomalonate ester 7.3
g of methanol (60 ml) at 0-5°C.
After adding caustic soda (80 ml) dropwise and stirring at room temperature for 16 hours, the reaction solution was concentrated under reduced pressure to about 80 ml, 60 ml of ethanol was added, the precipitated solid was collected, washed with 50 ml of ethanol, and dried in vacuum for 16 hours. , 6.0 g of the desired product (a) is obtained as a colorless powder having a melting point (decomposition point) of 254-255°C. Elemental analysis value C ₁₃ H ₉ N ₂ O ₅ ClNa ₂・2H ₂ O Calculated value: C, 39.97; H, 3.35; N, 7.19; Cl, 9.07 Actual value: C, 40.43; H, 3.43; N, 7.14 ;Cl, 9.21 and 2-(p-chlorobenzoylamino)-2
-(2′-cyanoethyl)disodium malonate 2
Add 50 ml of acetic acid ethyl ester to a solution of 4.0 g of aqueous salt in water (50 ml), and add 1N hydrochloric acid to PH = 2.0 at 0 to 5°C.
and dry the organic layer over magnesium sulfate. The residue obtained by distilling off the solvent is recrystallized from ethyl ether to obtain 3.2 g of the desired product (b) as a colorless powder having a melting point (decomposition point) of 103 to 105°C. Elemental analysis value C ₁₃ H ₁₁ N ₂ O ₅ As Cl Calculated value: C, 50.26; H, 3.57; N, 9.02; Cl, 11.41 Actual value: C, 49.84; H, 3.64; N, 9.03; Cl, 11.68 Same Amino acid derivatives exemplified below can be obtained by the method.

[Table] Formulation Example 1 Injection 2-(p-chlorobenzoylamino)-2-
10 mg of disodium (2'-cyanoethyl)malonate dihydrate was dissolved in 5 ml of physiological saline and sterilized by heat according to a conventional method to prepare an injection. Formulation Example 2 Oral capsule 2-(p-chlorobenzoylamino)-2-
(2'-cyanoethyl)malonate disodium dihydrate 50mg Lactose 100mg Corn starch 148.5mg Magnesium stearate 1.5mg Total 300mg After mixing the powder of the above formulation and passing it through a 30-mesh sieve, 300mg of this powder was It was put into gelatin capsules and made into capsules.

Claims (1)

[Claims] 1 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
A mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and A is an alkyl group that may have a hydroxyl group, an alkoxy group, an amino group, a halogen atom, or a carbamoyl group. Substituted amino group, hydrazino group, lower alkyl or aryl substituted hydrazino group,
Indicates a hydroxyamino group, lower alkoxyamino group, or aralkyloxyamino group. however,
When R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is any of the above other than a hydrogen atom, a lower alkyl group, a cyano group, a cyano-lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. Indicates the group of ) and its pharmacologically acceptable salts. 2 formulas (In the formula, R ₄ is a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group, mercapto lower alkyl group, alkylthio lower alkyl group) or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group,
M ₁ represents an alkyl group, and R ₂ and R ₃ are the same or different and represent a hydrogen atom or a lower alkyl group. ) is a compound with the formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or X represents a halogen atom. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ in the above formula represents a hydrogen atom, a lower alkyl group, a cyano group, or a cyano lower A formula characterized by reacting a compound having an alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or any of the above groups other than an aryl group in the presence of a base. A method for producing a compound represented by the formula (wherein R ₁ , R ₂ , R ₃ , R ₄ , n and M ₁ are the same as defined above). 3 formulas (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
It represents a mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and M ₁ represents an alkyl group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a group other than a hydrogen atom, a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. represents the above group. ) is characterized in that it hydrolyzes a compound having (In the formula, R ₁ , R ₂ , R ₃ , R ₄ and n are the same as defined above.) A method for producing a compound and a metal salt thereof. 4 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or _R4 is a hydrogen atom, a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, a carboxy group, an aryl group, an aralkyl group, a mercapto lower alkyl group,
It represents a disulfide group having a symmetrical structure corresponding to the alkylthio lower alkyl group or the mercapto lower alkyl group, and M ₁ represents an alkyl group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a hydrogen atom,
Indicates the above groups other than a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. ) in the presence of a strong base to form a compound with the formula (In the formula, R ₂ and R ₃ are the same or different and represent a hydrogen atom or a lower alkyl group.) A method for producing a compound represented by the formula (wherein R ₁ , R ₂ , R ₃ , R ₄ , n and M ₁ are the same as defined above). 5 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
It represents a mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a hydrogen atom,
Indicates the above groups other than a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. ) and a compound having the formula HM ₂ (wherein M ₂ is an amino group, an alkyl-substituted amino group optionally having a halogen atom or a carbamoyl group, a hydrazino group, a lower alkyl- or aryl-substituted hydrazino group, a hydroxyamino group, A formula characterized by reacting a compound having a lower alkoxyamino group or an aralkyloxyamino group. A method for producing a compound represented by the formula (wherein R ₁ , R ₂ , R ₃ , R ₄ , n and M ₂ are the same as defined above). 6 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group. ) is heated in the presence of a base. A method for producing a compound represented by the formula (wherein R ₁ , R ₂ , R ₃ and n are the same as defined above). 7 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
A mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and A is an alkyl group that may have a hydroxyl group, an alkoxy group, an amino group, a halogen atom, or a carbamoyl group. It represents a substituted amino group, hydrazino group, lower alkyl group or aryl substituted hydrazino group, hydroxyamino group, lower alkoxyamino group or aralkyloxyamino group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a group other than a hydrogen atom, a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. represents the above group. )
An immunostimulant comprising a compound represented by and a pharmacologically acceptable salt thereof as an active ingredient. 8 formula (In the formula, R ₁ is a hydrogen atom, a lower alkyl group, a halogeno-lower alkyl group, a halogen atom, a mercapto group, an alkylmercapto group, a hydroxyl group, an alkoxy group, an acyloxy group, a carboxy group, an alkoxycarbonyl group, an amino group, an alkyl-substituted amino group, aryl-substituted amino group, acylamino group, halogeno lower alkoxycarbonylamino group, alkanesulfonyl group, nitro group or cyano group, n represents 1 to 5, and when n represents 2 to 5, R ₁ is the same or May be different. _{R 2} and
R ₃ represents the same or different hydrogen atom or lower alkyl group, R ₄ represents a hydrogen atom, lower alkyl group, cyano group, cyano lower alkyl group, hydroxy lower alkyl group, lower alkoxycarbonyl group, carboxy group, aryl group, aralkyl group,
A mercapto lower alkyl group, an alkylthio lower alkyl group, or a disulfide group having a symmetrical structure corresponding to the mercapto lower alkyl group, and A is an alkyl group that may have a hydroxyl group, an alkoxy group, an amino group, a halogen atom, or a carbamoyl group. It represents a substituted amino group, hydrazino group, lower alkyl group or aryl substituted hydrazino group, hydroxyamino group, lower alkoxyamino group or aralkyloxyamino group. However, when R ₁ is a hydrogen atom, a lower alkyl group, or a halogen atom, R ₄ is a group other than a hydrogen atom, a lower alkyl group, a cyano group, a cyano lower alkyl group, a hydroxy lower alkyl group, a lower alkoxycarbonyl group, or an aryl group. represents the above group. )
An anticancer agent containing a compound represented by the formula and a pharmacologically acceptable salt thereof as an active ingredient.