JPH0672866A - Differentiation inducing agent - Google Patents

Abstract

PURPOSE: To provide a differentiation inducing agent having a specified compd. as an effective competent which is less toxic, excellent in safeness and effective in preventing growth of malignant tumor and curing malignant tumor. CONSTITUTION: This differentiation inducing agent contains a compd. (salt) expressed by formula I, such as 4-[(3,5-di-t-buty]-4-hydroxyphenyl)carboxyamide] benzoate as an effective component. In formula I, R<1> is H, 1 to 5C alkyl group, 2 to 5C alkanoyl group, X is vinylene, -COCH=CH-, -COO-, etc., and R<3> is expressed by formula II, wherein R<4> is H, halogen, cyano, carbamoyl, sulfoneamide group, etc., and R<5> is H, hydroxyl, nitro or amino group.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a differentiation inducer useful for the prevention and treatment of malignant tumor growth.

[0002]

2. Description of the Related Art Vitamin A or vitamin A acid has important physiological activity for maintaining normal cell growth and differentiation. Vitamin A acid and its analogs are used in the prevention and treatment of malignant tumor growth and in the treatment of acne, psoriasis and other skin disorders and can be used systemically or locally. German Patent Nos. 3434948 and 343494
2, European Patent No. 2210118 and H.M. K
agechika et al. Med. Chem, No. 31
Vol., 2182-2192 (1988), has two aromatic rings and is linked to an aromatic carboxylic acid or a carboxylic acid through a substituted vinylene group, an amide bond, an azo bond, a ketene bond or the like. It is described that the "bioelectron resonator" has the action of inducing cell differentiation and preventing and treating skin diseases. Also, European Patent No. 212
No. 848, No. 211548 and C. of Shudo et al.
hem. Pharm. Bull. , Vol. 34, 121
Page (1986) discloses a compound having a di-t-butylphenyl group, which is said to be effective for treating asthma, allergy, psoriasis and the like. However, the compounds that have been reported so far have not been clinically applied because they have insufficient effects such as induction of cell differentiation and are highly toxic such as vitamin A excess.

[0003]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a differentiation inducer having a superior action as compared with the conventional one.

[0004]

Means for Solving the Problems As a result of intensive studies in view of the above-mentioned objects, the present inventors have found that a series of known or newly synthesized compounds have an excellent differentiation-inducing action,
The present invention has been completed.

The present invention has the following formula [1]:

[0006]

[In the formula, R ¹ is a hydrogen atom or a carbon atom number 1 to
5 represents an alkyl group of 5 or an alkanoyl group having 2 to 5 carbon atoms, and X is a vinylene group or the following formula

[0008]

(Wherein R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms), and R ³ represents a group represented by the formula

[0010]

(In the formula, R ⁴ is a hydrogen atom, a halogen atom,
Cyano group, carbamoyl group, sulfonamide group, carboxyl group, alkoxycarbonyl group having 2 to 5 carbon atoms, alkyl group having 1 to 5 carbon atoms, 1 to 5 carbon atoms
Represents an alkoxy group, a trifluoromethyl group, an acetyl group or a tetrazolyl group, and R ⁵ is a hydrogen atom, a hydroxyl group,
Indicates a nitro group or an amino group. ] It is a differentiation inducer which uses the compound or its salt represented by these as an active ingredient.

Another aspect of the present invention is the following formula [2]:

[0013]

(In the formula, R ⁶ and R ⁷ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, Y is a single bond or a vinylene group, and Z is O or NH. ) Is a differentiation inducer containing a fused heterocyclic carboxylic acid derivative and a salt thereof represented by

In the present invention, the alkyl group is a linear or branched alkyl group such as a methyl group,
Examples thereof include ethyl group, propyl group, isopropyl group, butyl group, isobutyl group and pentyl group. The alkanoyl group is a linear or branched alkanoyl group, and examples thereof include an acetyl group, a propionyl group, an isopropionyl group and a butyryl group. The alkoxycarbonyl group includes a linear or branched alkyl group, and examples thereof include a methoxycarbonyl group, an ethoxycarbonyl group and a propoxycarbonyl group.

The salt of the compound according to the present invention is pharmacologically acceptable and is formed when R ⁴ is a carboxyl group in the formula [1] and when R is a hydrogen atom in the formula [2]. And an inorganic base such as potassium, sodium, magnesium, ammonia or a salt with an organic base such as triethylamine.

In the formula [1], when R ⁴ is a tetrazolyl group, there are two tautomers, and the compound according to the present invention also includes these tautomers.

Preferred compounds according to the present invention are 4-
[(3,5-Di-t-butyl-4-hydroxyphenyl) carboxamide] benzoic acid, 4-[(3,5-di-
t-Butyl-4-hydroxyphenyl) carboxamide] ethyl benzoate, 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxy] benzoic acid, 4
Ethyl-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxy] benzoate, 4- [3- (3,5
-Di-t-butyl-4-hydroxyphenyl) -3-oxo-1-propenyl] benzoic acid, 2- (3,5-di-
t-Butyl-4-hydroxyphenyl) benzimidazole-6-carboxylic acid, 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzimidazole-6-
Ethyl carboxylate, 2- (3,5-di-t-butyl-4)
-Methoxyphenyl) benzimidazole-6-carboxylic acid, ethyl 2- (3,5-di-t-butyl-4-methoxyphenyl) benzimidazole-6-carboxylate,
Ethyl 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzoxazole-5-carboxylate, 2-
(3,5-di-t-butyl-4-hydroxyphenyl)
Benzoxazole-5-carboxylic acid, 4-[(3,5
-Di-t-butyl-4-hydroxyphenyl) carboxamide] -3-nitrobenzoic acid, 4-[(3,5-di-
t-Butyl-4-hydroxyphenyl) carboxamide] -3-nitrobenzoic acid ethyl ester, 3-amino-4-
[(3,5-Di-t-butyl-4-hydroxyphenyl) carboxamide] methyl benzoate, 3-amino-
4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamide] benzoic acid.

The compounds according to the present invention include some known compounds, and each new compound can be produced by the following method.

(1-1): First, the compound of the formula [1] in which X is a group represented by the formula -CON (R ² )-(wherein R ² has the same meaning as described above) is , Formula [3]

[0021]

(Wherein R ¹ has the same meaning as defined above) and a carboxylic acid derivative of the formula [4]

[0023]

An aniline derivative represented by the formula (wherein R ⁸ is R ⁴ excluding a carboxyl group and R ⁹ is R ⁵ excluding an amino group) can be prepared as a starting material.

That is, the carboxylic acid derivative of the above formula [3] is converted into an acid halide with a halogenating agent, and then this is reacted with the aniline derivative of the formula [4] in the presence of a base to obtain the formula [5].

[0026]

A compound represented by the formula (wherein R ¹ , R ⁸ and R ⁹ are as defined above) can be obtained. Here, the halogenating agent is a customary one using an organic carboxylic acid as an acid halide, and examples thereof include thionyl chloride and phosphorus oxychloride. As the base, both an inorganic base and an organic base are used, and they are, for example, potassium carbonate, pyridine, triethylamine and the like.

(1-2): The compound of the formula [5] has the formula R ¹⁰
A halogenated alkyl represented by —X ′ (in the formula, R ⁶ is an alkyl group having 1 to 5 carbon atoms represented by R ² , and X ′ is a chlorine atom, a bromine atom or an iodine atom). By reacting with the formula [6]

[0029]

A compound represented by the formula (wherein R ¹ , R ⁸ , R ⁹ and R ¹⁰ are as defined above) can be obtained. In this reaction, as the base, both an inorganic base and an organic base are used, and they are, for example, sodium hydroxide,
Examples include potassium hydroxide, potassium carbonate, sodium hydride, pyridine and triethylamine.

(1-3): Among the compounds of the formulas [5] and [6], the compound in which R ⁸ is an alkoxycarbonyl group can be converted into a carboxyl group by hydrolyzing the alkoxycarbonyl group. The hydrolysis is easily carried out by the usual method of hydrolyzing an ester.
For example, there is a method using alkali hydroxide and hydrous alcohol.

(1-4): R ¹ has 2 to 2 carbon atoms.
The compound of the formula [1] which is the alkanoyl group of 5 is 3,5
-Di-t-butyl-4-hydroxybenzoic acid and the formula [4]
Of the aniline derivative compound obtained by reacting in the same manner as (1-1), wherein R ¹¹ ₂ O (wherein, R ¹¹ is an alkanoyl group having 2 to 5 carbon atoms represented by R ¹ It can also be obtained by reacting with an acid anhydride represented by

(1-5): The compound of the formula [1] in which R ⁴ is a carboxyl group is esterified with an alcohol having 1 to ⁴ carbon atoms so that R ⁴ has 2 to 5 carbon atoms. A compound according to the present invention which is an alkoxycarbonyl group of

(2) The compound of the formula [1] in which X is a group represented by the formula —COO— has the following formula [7] instead of the aniline derivative of the formula [4] in the above (1-1).

[0035]

(In the formula, R ⁸ and R ⁹ have the same meanings as above.) The phenol derivative can be obtained by reacting in the same manner as in (1-1). The hydrolysis is also the above (1-
It can be performed in the same manner as 3).

(3) X is of the formula --N (R ² ) CO--

The compound of the formula [1], which is a group represented by the formula (wherein R ² has the same meaning as described above), is a compound of the formula [8]

[0039]

(In the formula, R ¹ has the same meaning as defined above) and the aniline derivative represented by the formula [9].

[0041]

(In the formula, R ⁸ and R ⁹ have the same meanings as described above.) Using the carboxylic acid derivative represented by the above as a starting material, the reaction is carried out in the same manner as in each reaction described in the above (1) to give The compounds according to the invention can be prepared.

(4) The compound of the formula [1] in which X is a group represented by the formula -COCH = CH- is a compound of the formula [10]

[0044]

(Wherein R ¹ has the same meaning as described above) and an acetophenonic acid derivative represented by the following formula [11]:

[0046]

The compound can be produced by reacting a formyl derivative represented by the formula (wherein R ⁸ and R ⁹ are as defined above) in the presence of barium hydroxide.

(5) The compound of the formula [1] in which X is a group represented by the formula —CH═CHCON (R ² ) — (wherein R ² has the same meaning as described above) is a compound of the formula [1] 12]

[0049]

The cinnamic acid derivative represented by the formula (wherein R ¹ has the same meaning as described above) and the aniline derivative of the formula [4] can be used as starting materials.

That is, by activating the carboxyl group of the cinnamic acid derivative of the above formula [11] with oxalyl chloride and then reacting this with the aniline derivative of the formula [4] in the presence of a base, the formula [13]

[0052]

The compound according to the present invention represented by the formula (wherein R ¹ , R ⁸ and R ⁹ have the same meanings as described above) can be obtained. As the base, both an inorganic base and an organic base are used, and they are, for example, potassium carbonate, pyridine, triethylamine and the like.

The N-alkylation of the compound of formula [13] is
It can be performed in the same manner as the method described in (1-2) above. Hydrolysis can be performed in the same manner as in the above (1-3).

(6) The compound of the formula [1] in which X is a group of the formula —N (R ² ) COCH═CH— (wherein R ² has the same meaning as described above) is An aniline derivative of [8] and the following formula [14]

[0056]

(Wherein R ⁸ and R ⁹ have the same meanings as described above), and a cinnamic acid derivative represented by
The compound according to the present invention can be produced in the same manner as each reaction described in (1) above.

(7) The compound of the present invention represented by the formula [2] is
When Z is O, the following formula [15]

[0059]

(Wherein Y has the same meaning as defined above) and the following formula [16]:

[0061]

It can be produced by heating an aniline derivative represented by the formula (R ¹⁰ is R ⁷ excluding hydrogen atom) in an organic solvent in the presence or absence of boric acid. . The organic solvent used in this reaction preferably has a relatively high boiling point, such as toluene or xylene. The completion of the reaction can be determined by the disappearance of the raw materials by thin layer silica gel chromatography or the like.

(8) Further, the compound of the present invention of the formula [2] in which Z is NH is the carboxylic acid derivative of the formula [15] and the following formula [17]:

[0064]

An aniline derivative represented by the formula (wherein R ¹⁰ has the same meaning as described above) can be used as a starting material. That is, the carboxylic acid derivative of the formula [15] is converted into an acid halide with a halogenating agent such as thionyl chloride, and then the aniline derivative of the formula [17] is reacted to form an amide. Then, the nitro group of the amide is reduced by catalytic reduction or the like, and the ring is closed by heating to obtain the desired product.

(9) The compound of the present invention represented by the formula [2] in which R ⁷ is a hydrogen atom is obtained by hydrolyzing the compounds obtained in the above (7) and (8) in the same manner as in the above (1-3). It can be obtained by disassembling.

[0067]

INDUSTRIAL APPLICABILITY The compound according to the present invention has a differentiation inducing action and is therefore useful for the prevention and treatment of malignant tumor growth.

Test Example 1 The differentiation inducing activity of the compound of the present invention was evaluated by using the differentiation inducing test of HL-60 cells. Collins et al.'S method (C
anser. Res. , Volume 42, 445-449
P., 1982). HL-60 cells 1 ×
10 ⁵ cells / ml RPMI-1 supplemented with 10% fetal bovine serum
The cells were cultured in 640 medium, penicillin 100 U / ml, streptomycin 100 μg / ml, 37 ° C., 5% CO ₂ . Compound 4- [3- (3,5-di-t-] according to the present invention
Butyl-4-hydroxyphenyl) -3-oxo-1-
Propenyl] benzoic acid and all-trans vitamin A
Acids were 1% and 0.1% ethanol solutions, and 0.1% ethanol solution was used as a control. At the start of culturing, drugs were added to each concentration to cultivate the cells for 5 days, and the differentiation-inducing activity was measured using the NBT reducing ability as an index. That is, after the culture was completed, the cells were collected by centrifugation at 1000 rpm, 0.5 ml of 0.1% NBT solution and 200 ng of 12-O-tetradecanoylphorbol-13-acetate (TPA) were added, and the mixture was incubated at 37 ° C. The reaction was carried out for 60 minutes. The cells after the reaction were collected by centrifugation, a smear was prepared, and stained with Wright-Giemsa stain. Observe 200 cells per specimen under the microscope,
If there is blue-purple formazan precipitate in the cytoplasm, N
The cells positive for BT reducing ability were selected.

As a result, 100% of the cells in the negative control group were immature cells, and the differentiation rate at 1 × 10 ⁻⁶ M was 9 in the positive control vitamin A acid.
5 to 100%, and the 50% differentiation rate (EC ₅₀ ) of the compound of the present invention was 7.1 × 10 ⁻⁷ M.

Test Example 2 The effect of the compound of the present invention on the expression of oncogene was
It was measured by the n-vitro method. Known method (Y. Lu
et al, J .; Biol. Chem. , Volume 263,
Pp. 4891-4894, 1988) under sterile conditions RPMI-1640 medium supplemented with 10% fetal bovine serum, penicillin 100 U / ml, streptomycin 1
HL- cultured at 37 ° C. at 00 μg / ml, 5% CO ₂ .
Different concentrations of compound 4- [3-
(3,5-di-t-butyl-4-hydroxyphenyl)
Treated with -3-oxo-1-propenyl] benzoic acid.
Cells were collected after 2, 6 and 12 hours, and the cells were collected in a guanidine isothiocyanate (GIT) solution [6
M guanidine isothiocyanate, 10 mM sodium citrate (pH 7), 0.5% sarcosyl, 0.1M2-
Mercaptoethanol] and shaken to disrupt cells. 5.7 M Cesium chloride-0.1 MEDTA-N
a (pH 7.5) was overlaid with the cell lysate, and
Density gradient centrifugation was performed at 4000 G and 20 ° C. for 16 hours. The colorless and transparent precipitate was used as total RNA, which was dissolved in water treated with DEPC, denatured at 65 ° C, and then Oligo-d.
OD (260n in buffer solution through T-cellulose column)
After washing until m) became 0, mRNA was eluted.
It was precipitated with ethanol and then denatured with DMSO,
1% agarose gel electrophoresis was performed and mRNA was immobilized on the nylon membrane by Northern transfer method. The DNA probe was labeled by nick translation. ^{A 32} P-labeled probe was bound to mRNA, and the oncogene expression state was measured by autoradiography.

The results are shown in FIG. It was found that the compound according to the present invention remarkably suppresses oncogene expression as compared with the control.

Test Example 3 The antimutagenic effect of the compound of the present invention was evaluated by a micronucleus formation test using mouse bone marrow cells. Micronucleus formation was induced by cyclophosphamide and DMBA, and the antagonism of the compound according to the present invention was observed. A group of 5 mice having a body weight of 18 to 22 g was prepared, and the compound 4- [3-
(3,5-di-t-butyl-4-hydroxyphenyl)
-3-Oxo-1-propenyl] benzoic acid 50 mg / k
g or 100 mg / kg was orally administered once a day for 4 consecutive days. After the completion of the administration on the 4th day, 100 mg / kg of cyclophosphamide or 25 mg / kg of DMBA was intraperitoneally injected to induce micronucleus formation, and 24 hours later, a micronucleus test was conducted by a conventional method. As shown in Tables 1 and 2, the compound according to the present invention significantly suppressed micronucleus formation as compared with the positive control group.

[Table 1]

[0073]

[0074]

[Table 2]

[0075]

Test Example 4 Method of Hecker et al. (Naturemissensa)
Feten, 54, 282, 1967).

The antipromoter activity of the compounds of the present invention was evaluated using a mouse ear swelling model with croton oil. Mice having a body weight of 18 to 22 g were randomly divided into groups, and each group contained 7 mice. Compound 4- [3- according to the present invention
(3,5-di-t-butyl-4-hydroxyphenyl)
-3-Oxo-1-propenyl] benzoic acid 50 mg / k
Oral oral administration of g or 100 mg / kg once a day for 4 days was performed, and after applying croton oil, holes were opened in both ears. The difference in the swelling between the applied ear and the unapplied ear was used as an index.

As a result, the difference in swelling between the ears of the control group was 2
5.3 ± 3.4 mg (P <0.05), 20.8 mg ± 4.8 mg (50 mg in the case of the compound group according to the present invention)
/ Kg) and 19.9 ± 3.9 mg (100 mg / kg)
(P <0.05). Therefore, it was determined that the compound according to the present invention has an inhibitory effect on ear swelling in mice.

Test Example 5 The effect of the compound of the present invention on tumor growth was evaluated using a rat chondrosarcoma model.

Chondrosarcoma (1: 3 dilution, 0.5 ml /
Animal) is subcutaneously inoculated into a rat, and then the compound 4- [3- (3,5-di-t-butyl-4-hydroxyphenyl) -3-oxo-invention is directly administered to the stomach every two days. 1-propenyl] benzoic acid 5 mg / kg or 10 mg / kg
Was cast continuously for 9 weeks. As a result of measuring the tumor weight of the rat at 7th week, the tumor suppression rates of 5 mg / kg and 10 mg / kg were 98.7% and 100%, respectively, relative to the control group. This result showed that the compound according to the present invention has a remarkable inhibitory effect on rat transplantable chondrosarcoma.

Test Example 6 The dermal antagonist of the compound of the present invention against chemical carcinogens was evaluated by using mouse skin papilloma caused by DMBA and croton oil as a model.

Mice having a body weight of 18 to 22 g were randomly divided into 30 groups of 30 mice, and their backs were hair-removed with an 8% aqueous sodium sulfide solution. After breeding for 1 week, hair was removed 1 time at a time
0.2 ml of a dimethylbenzanthracene-acetone solution containing 50 ng DMBA was applied and administered twice a week for 2 weeks. From the 3rd week, a 0.25% croton oil-acetone solution was applied in an amount of 0.2 ml / dose and administered twice a week for 12 weeks. 50 mg / k from the first week when DMBA is applied
g, 25 mg / kg Compound 4- [3-
(3,5-di-t-butyl-4-hydroxyphenyl)
-3-Oxo-1-propenyl] benzoic acid was administered directly to the back of the mice. Compounds according to the invention 1 every 2 days
It was administered once every 12 weeks, and the number of animals in which papilloma developed and the number of tumors in each animal were recorded for each week.

As a result, the compound of the present invention had a remarkable inhibitory action as compared with the control, and its activity was dose-dependent (FIG. 2).

[0084]

EXAMPLES The present invention will be described in more detail with reference to Production Examples and Examples of the compounds according to the present invention.

Production Example 1 3,5-di-t-butyl-4-hydroxybenzoic acid 50
g (O.2mol) and thionyl chloride 50g (0.42m
ol) was heated under reflux for 30 minutes with stirring to give a clear pale yellow solution. Remove the acidic gas generated under reduced pressure,
Petroleum ether was added, and vacuum distillation was performed again. This 2
By repeating this, pale yellow crystals were obtained. p-
Ethyl aminobenzoate 33.04 g (O.2 mol) was dissolved in 500 ml of anhydrous ether to give anhydrous pyridine 16
g (0.2 mol) was added to make a colorless homogeneous solution, and the compound obtained by the above reaction was added rapidly with vigorous stirring, and the mixture was stirred until it became homogeneous, and a white precipitate soon precipitated. The mixture was further heated under reflux for 4 hours with stirring, and left overnight at room temperature.

The ether was distilled off under reduced pressure, petroleum ether was added to the residual white solid, the mixture was filtered through a Nutsche and washed with petroleum ether. After drying, wash the pyridine hydrochloride off with water,
It is dried again and the yield of 100% is 4-[(3,5-di-t-
Butyl-4-hydroxyphenyl) carboxamide] ethyl benzoate was obtained. mp 274-277 ° C (recrystallized with ethanol)

Production Example 2 The crude product of the compound produced in Production Example 1 was mixed with 400 ml of 95% ethanol and 20 g of sodium hydroxide (0.5 mol).
It was suspended in a mixed solution with 100 ml of an aqueous solution containing 1) and stirred at room temperature for 8 to 10 hours, and when the solution became a uniform transparent solution, the mixture was left overnight at room temperature. At room temperature, excess ethanol was distilled off under reduced pressure, 200 ml of water was added to the remaining liquid, and the filtrate was filtered under reduced pressure. It was It is dried and recrystallized from dilute ethanol to give 4-[(3,5-di-t
-Butyl-4-hydroxyphenyl) carboxamide]
66.1 g of benzoic acid was obtained. mp 260-263 ° C

Production Example 3 Ethyl 4-hydroxybenzoate 16.6 g (0.1 mo)
4-[(3,5-di-t-butyl-4) in the same manner as in Production Example 1 and then Production Example 2 using 1) as a raw material.
-Hydroxyphenyl) carboxy] benzoic acid 28.0
g was obtained. mp 218-220 ° C

Production Example 4 3,5-di-t-butyl-4-hydroxybenzoic acid 2.
50 g (10 mmol) and aniline 0.93 g (10 m
mol) as a raw material and by the production method of Production Example 1, 2.0 g of N-phenyl-3,5-di-t-butyl-4-hydroxybenzamide was obtained. mp 218-220 ° C

Production Example 5 3,5-di-t-butyl-4-hydroxybenzoic acid
50 g (10 mmol) and 4-chloroaniline 1.27
3.50 g of N- (4-chlorophenyl) -3,5-di-t-butyl-4-hydroxybenzamide by the production method of Production Example 1 using g (10 mmol) as a raw material.
Got mp 264-226 ° C

Production Example 6 3,5-di-t-butyl-4-hydroxybenzoic acid
Using 50 g (10 mmol) and 4-aminobenzenesulfonamide 1.72 g (10 mmol) as raw materials,
By the production method of Production Example 1, 2.45 g of N- (4-sulfonamidophenyl) -3,5-di-t-butyl-4-hydroxybenzamide was obtained. mp 272-274 ° C

Production Example 7 3,5-di-t-butyl-4-hydroxybenzoic acid 2.
50 g (10 mmol) and 4-methoxyaniline 1.2
By using 3 g (10 mmol) as a raw material and the production method of Production Example 1, N- (4-methoxyphenyl) -3,5
-Di-t-butyl-4-hydroxybenzamide 2.5
6 g was obtained. mp 224-226 ° C

Production Example 8 3,5-di-t-butyl-4-hydroxybenzoic acid
50 g (10 mmol) and P-methylaniline 1.07
2.94 g of N- (4-methylphenyl) -3,5-di-t-butyl-4-hydroxybenzamide was produced by the production method of Production Example 1 using g (10 mmol) as a raw material.
Got mp 228-230 ° C

Production Example 9 3,5-di-t-butyl-4-hydroxybenzoic acid
Using 50 g (10 mmol) and 1.36 g (10 mmol) of 4-aminobenzoylamide as raw materials, N- (4-carbamoylphenyl) -3,5-di-t-butyl-4- was prepared according to the production method of Production Example 1. 3.10 g of hydroxybenzamide was obtained. mp 307-309 ° C

Production Example 10 3,5-di-t-butyl-4-hydroxybenzoic acid 2.
50 g (10 mmol) and 4-acetylaniline 1.3
N- (4-acetylphenyl) -3,5 was obtained by the production method of Production Example 1 using 5 g (10 mmol) as a raw material.
-Di-t-butyl-4-hydroxybenzamide 3.0
0 g was obtained. mp 265-268 ° C

Production Example 11 3,5-di-t-butyl-4-hydroxybenzoic acid 2.
50 g (10 mmol) and 3-aminobenzoic acid 2.34
3-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamido] benzoic acid 4.80 was obtained by the production method of Production Example 1 using g (10 mmol) as a raw material.
g was obtained. mp 273-275 ° C

Production Example 12 3,5-di-t-butyl-4-hydroxybenzoic acid 10
g [40 mmol] and 6.65 g (40 mmol) of ethyl 3-hydroxybenzoate as raw materials were used to prepare 3-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxy by the production method of Production Example 1. ] 14.20 g of ethyl benzoate was obtained. mp 110-112 ° C

Production Example 13 12 g of the compound obtained in Production Example 12 by the method of Production Example 2
Hydrolyze (30 mmol) to give 3-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxy]
10.50 g of benzoic acid was obtained. mp 211-213 ° C

Production Example 14 3,5-di-t-butyl-4-hydroxybenzoic acid 1.
Using 02 g (6.3 mmol) and 4-trifluoromethylbenzoic acid as raw materials, according to the production method of Production Example 1,
1.85 g of N- (4-trifluoromethylphenyl) -3,5-di-t-butyl-4-hydroxybenzamide
Got mp 234-236 ° C

Production Example 15 3,5-di-t-butyl-4-hydroxybenzoic acid 2.
50 g (10 mmol) and 4-aminosalicylic acid 1.5
4-[(3,5-di-t-butyl-4-] was obtained by the production method of Production Example 1 using 3 g (10 mmol) as a raw material.
Hydroxyphenyl) carboxamide] -2-hydroxybenzoic acid (2.80 g) was obtained. mp 246-248 ° C

Production Example 16 According to the production method of Production Example 1, 3,5-di-t-butyl-
2.50 g (10 mmol) of 4-hydroxybenzoic acid and 1.96 g of methyl 3-nitro-4-aminobenzoate (1
(0 mmol) was reacted and recrystallized from ethanol to obtain 1.04 g of methyl 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamido] -3-nitrobenzoate as yellow crystals. It was mp 166-168 ° C

Production Example 17 By the method of Production Example 2, the compound 0.8 obtained in Production Example 16 was used.
Hydrolyze g (1.87 mmol) to give 4-[(3,
0.63 g of 5-di-t-butyl-4-hydroxyphenyl) carboxamide] -3-nitrobenzoic acid was obtained. mp 266-268 ° C

Production Example 18 1.0 g (2.3 mmol) of methyl 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamide] -3-nitrobenzoate was suspended in 30 ml of 95% ethanol. Make it turbid, add a small amount of Raney nickel, and add Parr
Reduced in hydrogenator. When the generation of hydrogen stopped, nickel was removed by filtration, the filtrate was evaporated to dryness under reduced pressure, and the obtained white product was recrystallized from ethanol to give 3-amino-4-[(3,5-di-t). 0.7 g of methyl-butyl-4-hydroxyphenyl) carboxamide] nitrobenzoate was obtained. mp 242-244 ° C

Production Example 19 The compound 0.8 obtained in Production Example 18 was prepared by the method of Production Example 2.
g (1.87 mmol) was hydrolyzed to give 3-amino-
0.63 g of 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamide] nitrobenzoic acid was obtained. mp 309-311 ° C

Production Example 20 0.2 g of the compound obtained in Production Example 18 and 0.02 g of p-toluenesulfonic acid were dissolved in 20 ml of xylene and heated under reflux for 20 hours. Xylene was distilled under reduced pressure, ethyl acetate was added to the residue, and the mixture was washed successively with a dilute aqueous sodium carbonate solution and water. After drying over anhydrous sodium sulfate, ethyl acetate was distilled off under reduced pressure to obtain 0.14 g of methyl 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzimidazole-6-carboxylate. mp 242-244 ° C

Production Example 21 0.8 g (1.87 mmol) of the compound obtained in Production Example 20
Was dissolved in 12 ml of ethanol, 3 ml of an aqueous solution containing 630 mg of sodium hydroxide was added, and the mixture was left overnight with stirring at room temperature. The reaction solution was neutralized to pH 7 with hydrochloric acid, most of ethanol was distilled off under reduced pressure, the pH was adjusted to 3 with hydrochloric acid, and 20 ml of water was added. The white solid thus formed was collected by filtration, crystallized from ethanol to give 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzimidazole-
0.63 g of 6-carboxylic acid was obtained. mp 302-305 ° C

Production Example 22 3,5-di-t-butyl-4-hydroxybenzoic acid 5.
35 g (21.6 mmol), ethyl 3-amino-4-hydroxybenzoate (3.9 g, 21.6 mmol) and boric acid (1.33 g, 21.6 mmol) were added to xylene 2.
It was added to 20 ml and heated to reflux with stirring for 120 hours (check occasionally by thin-layer silica gel chromatography until no more raw material). Xylene was distilled off under reduced pressure,
Add 10 parts each of saturated aqueous sodium carbonate and water to the black residue.
Add 0 ml and extract with methylene chloride (4 x 150 m
l). After combining the organic layers and washing with water until neutral,
It was dried over anhydrous magnesium sulfate, and methylene chloride was distilled off under reduced pressure to obtain a black residue. Silica gel column chromatography, eluting with methylene chloride, 2-
(3,5-di-t-butyl-4-hydroxyphenyl)
5.0 g of ethyl benzoxazole-5-carboxylate was obtained.

Mp 156 to 157 ° C. Elemental analysis: C ₂₄ H ₂₉ NO ₄ = 395.48 Calculated value (%): C 72.87, H 7.39, N
3.55 Found (%): C 72.99, H 7.39, N
3.49 ¹ H-NMR (CDCl ₃ ) δppm: 1.39 (5,
_{3H, -CH 3), 1.45 (} s, 18H, t-B
u), 4.38 (q, 2H , -CH 2 -), 7.43-
8.38 (m, 5H, Ar-H) MS m / e: 395 (M ⁺ , 93), 380 (M ⁺ -C).
H ₃ , 100) IR (KBr) ν (cm ^-1 ): 3600, 2980,
2940, 1715, 1620, 1550, 1440,
1420, 1370, 1280, 1250, 810, 7
Fifty

Production Example 23 In the same manner as in Production Example 21, the compound 34 obtained in Production Example 22 was used.
Hydrolyze 0 mg (0.86 mmol) to give 2-
(3,5-di-t-butyl-4-hydroxyphenyl)
260 mg of benzoxazole-5-carboxylic acid was obtained.

Mp 305 to 307 ° C. (recrystallized from ethanol) Elemental analysis: C ₂₂ H ₂₅ NO ₄ = 367.44 Calculated value (%): C 71.91, H 6.86, N
3.81 Found (%): C 71.46, H 6.81, N
^{4.35 1 H-NMR (DMSO-} d 6) δppm: 1.43
(S, 18H, t-Bu), 7.77-8.19 (m,
5H, Ar-H) MS m / e: 367 (M ⁺ , 56), 352 (MC)
H ₃ , 100) IR (KBr) ν (cm ⁻¹ ): 3620, 2980,
1680, 1620, 1550, 1500, 1420,
1360, 1300, 1240, 830, 770, 75
0

Production Example 24 As in Production Example 21, 3.80 g (15.2 mmol)
2- (3,5-di-t-butyl-4-hydroxyphenyl) from 3,5-di-t-butyl-4-hydroxybenzoic acid and 2.50 g of ethyl 3-hydroxy-4-aminobenzoate. ) 1.20 g of ethyl benzoxazole-6-carboxylate was obtained.

Mp 138-139 ° C. ¹ H-NMR (CDCl ₃ ) δppm: 1.40 (t,
_{3H, -CH 3), 1.48 (} s, 18H, t-B
u), 4.40 (q, 2H , -OCH 2 -), 7.22
-8.22 (m, 6H, Ar-H) MS m / e: 395 (M ⁺ , 87), 380 (M ⁺ -C).
H ₃ , 100)

Production Example 25 In the same manner as in Production Example 22, the compound 1.
Hydrolyze 0 g (2.53 mmol) to give 2- (3,
0.42 g of 5-di-t-butyl-4-hydroxyphenyl) benzoxazole-6-carboxylic acid was obtained.

Mp 308 to 310 ° C. (recrystallized from ethanol) Elemental analysis: C ₂₂ H ₂₅ NO ₄ = 367.44 Calculated value (%): C 71.91, H 6.86, N
3.81 Found (%): C 71.85, H 6.87, N
4.10 ¹ H-NMR (DMSO-d ₆ ) δppm: 1.44
(S, 18H, t-Bu), 7.76-8.22 (m,
5H, Ar-H) MS m / e: 367 (M ⁺ , 70), 352 (M ⁺ -C).
H ₃ , 100) IR (KBr) ν (cm ^-1 ): 3620, 2960,
1680, 1610, 1550, 1420, 1360,
1300, 1240, 1150, 890, 770

Production Example 26 In the same manner as in Production Example 21, 1.22 g (4.41 mmol)
3,5-di-t-butyl-4-hydroxycinnamic acid and 0.80 g (4.41 mmol) 3-amino-4
From ethyl -hydroxybenzoate, 2- [2- (3,5
Ethyl] -di-t-butyl-4-hydroxyphenyl) ethenyl] benzoxazole-5-carboxylate 0.8
0 g was obtained.

Mp 117 to 118 ° C. (recrystallized from isopropyl ether) Elemental analysis: C ₂₆ H ₃₁ NO ₄ = 421.52 Calculated value (%): C 74.08, H 7.41, N
3.34 Found (%): C 74.27, H 7.44, N
_{^{3.44 1 H-NMR (CDCl 3}} ) δppm: 1.39 (t,
_{3H, -CH 3), 1.43 (} s, 18H, t-B
u), 4.18 (q, 2H , -OCH 2 -), 7.38
-8.30 (m, 5H, Ar-H) MS m / e: 421 (M ⁺ , 100), 406 (M-
CH ₃ , 59) IR (KBr) ν (cm ^-1 ): 3600, 2980,
1720, 1630, 1600, 1530, 1290,
840

Production Example 27 The compound 30 obtained in Production Example 26 was prepared in the same manner as in Production Example 22.
Hydrolyze 0 mg (0.71 mmol) to give 2- [2
-(3,5-Di-t-butyl-4-hydroxyphenyl) ethenyl] benzoxazole-5-carboxylic acid 2
Obtained 00 mg.

[0118]

Mp 176 to 177 ° C. (recrystallized from ethanol) Elemental analysis: C ₂₄ H ₂₇ NO ₄ = 393.48 Calculated value (%): C 73.25, H 6.92, N
3.57 Found (%): C 73.20, H 7.08, N
3.41 ¹ H-NMR (DMSO-d ₆ ) δppm: 1.40
(S, 18H, t-Bu), 7.40-8.04 (m,
5H, Ar-H) MS m / e: 393 (M ⁺ , 13), 219 (10) IR (KBr) ν (cm ^-1 ): 3610, 3420,
3030, 2980, 1700, 1640, 1600,
1530, 1430, 1360, 1260, 1200,
1100,890,750

Production Example 28 3,5-di-t-butyl-4-methoxybenzoic acid 1.2
0 g (4.54 mmol) was dissolved in 15 ml of benzene, 5 ml of thionyl chloride was added, and the mixture was heated under reflux for 3 hours.
Benzene and excess thionyl chloride were distilled off under reduced pressure, and the residue was dissolved in 30 ml of anhydrous ether. To this, 0.75 g (4.54 mmol) of ethyl 4-aminobenzoate and 10 ml of anhydrous pyridine were added, and the mixture was stirred at 40 ° C. for 3 hours. The ether was removed under reduced pressure, 30 ml of water was added to the residue, the pH was adjusted to 4 with hydrochloric acid, the mixture was extracted 4 times with 40 ml of ether, and the ether layers were combined and washed successively with saturated saline solution and water, and anhydrous sodium sulfate. After drying with, the ether was distilled off to obtain 1.80 g of a pale yellow solid. This was recrystallized from ethanol and 4-[(3,5-di-t-
Butyl-4-methoxyphenyl) carboxamido] ethyl benzoate (1.5 g) was obtained.

Mp 171-172 ° C. Elemental analysis: C ₂₅ H ₃₃ NO ₄ = 411.52 Calculated value (%): C 72.96, H 8.08, N
3.40 Found (%): C 72.90, H 7.97, N
3.28 ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.43
(S, 18H, t-Bu), 1.40 (t, 3H, -C)
_{H 3), 3.72 (s,} 3H, -OCH 3), 4.38
_{(Q, 2H, -OCH 2 -} ), 7.60-8.08
(M, 6H, Ar-H) MS m / e: 412 (M ⁺ +1,6), 247 (10
0) IR ν (cm ^-1 ): 3220, 3100, 2950,
1715, 1645, 1595, 1530, 1265,
1220,770

Production Example 29 0.79 g (1.92 mmo) of the compound obtained in Production Example 28
l) was dissolved in 12 ml of ethanol, 3 ml of an aqueous solution containing 640 mg of sodium hydroxide was added, and the mixture was left overnight with stirring at room temperature. The reaction solution was neutralized to pH 7 with hydrochloric acid, most of ethanol was distilled off under reduced pressure, and the pH was further adjusted with hydrochloric acid.
Adjusted to 3 and added 20 ml of water. The produced white solid was collected by filtration and crystallized from ethanol to obtain 0.53 g of 4-[(3,5-di-t-butyl-4-methoxyphenyl) carboxamido] benzoic acid.

Mp 285-286 ° C. Elemental analysis: C ₂₃ H ₂₉ NO ₄ = 383.47 Calculated value (%): C 72.04, H 7.62, N
3.65 Found (%): C 72.22, H 7.65, N
4.01 ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.44
(S, 18H, t-Bu), 3.69 (s, 3H, -O)
_{CH 3), 7.80-7.90 (m,} 6H, Ar-
H), 10.38 (s, 1H , -CO 2 H) MS m / e: 383 (M +, 16), 366 (M-O
H, 7), 247 (100) IR (KBr) ν (cm ^-1 ): 3200, 2960,
1685, 1645, 1600, 1525, 1400,
1220,780

Production Example 30 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamido] benzoic acid 1.0 g (2.71 mmol) produced in Production Example 2 was mixed with 3 ml of acetic anhydride. Then, the mixture was heated with stirring and dissolved until it became transparent, whereby a white solid was deposited. After cooling, ice-cold water was added to the reaction solution, the solid was collected by filtration, washed with water to be neutral, dried and crystallized with ethyl acetate, and then 4-[(4-acetoxy-3,
5-di-t-butylphenyl) carboxamide] benzoic acid 1.05 g was obtained. mp 223-226 ° C MS m / e: 412 (M ⁺ +1,10), 351 (M ⁺
-60, 6), 233 (100)

Production Example 31 2 g of powdery potassium hydroxide was added to dimethyl sulfoxide 2
It was mixed with 0 ml and stirred at room temperature for 5 minutes. In addition to this, 4-[(3,5-di-t-butyl-4-
Hydroxyphenyl) carboxamide] benzoic acid 2.0
g (5.4 mmol) and 1 ml of methyl iodide (16
mmol) was added and the mixture was stirred at room temperature for 1 hour. 6 reaction liquids
Pour into 0 ml water, add salt to saturate, and add hydrochloric acid to adjust pH.
The mixture was adjusted to 3 and extracted 3 times with 50 ml of ether, and the ether layers were combined and washed with a saturated sodium chloride solution until neutral and then with water. The ether layer was dried over magnesium sulfate, and the ether was distilled off under reduced pressure to obtain 2.0 g of a yellow solid. This was crystallized with ethanol, decolorized with activated carbon, and
Methyl-[N-methyl (3,5-di-t-butyl-4-methoxyphenyl) carboxamido] benzoate was 1.7.
g was obtained.

Mp 126-127 ° C. Elemental analysis: C ₂₅ H ₃₃ NO ₄ = 411.52 Calculated value (%): C 72.96, H 8.08, N
3.40 Found (%): C 72.63, H 7.87, N
3.47 ¹ H-NMR (CDCl ₃ ) δ (ppm): 1.25
(S, 18H, t-Bu), 3.52 (s, 3H, -N)
_{CH 3), 3.59 (s,} 3H, -OCH 3), 3.88
_{(S, 3H, -CO 2 CH} 3), 7.00-7.92
(M, 6H, Ar-H) MS m / e: 411 (M ⁺ , 6.7), 247 (10
0) IR (KBr) ν (cm ⁻¹ ): 2950, 1720,
1600, 1570, 1430, 1330, 1270,
1220, 1170, 1110, 860, 770

Production Example 32 According to the method of Production Example 29, the compound 1.
0 g was hydrolyzed to give 4- [N-methyl (3,5-di-t
0.8 g of -butyl-4-methoxyphenyl) carboxamide] benzoic acid was obtained.

Mp 173-174 ° C. (recrystallized from ethanol) Elemental analysis: C ₂₄ H ₃₁ NO ₄ = 397.50 Calculated value (%): C 72.52, H 7.86, N
3.54 Found (%): C 72.48, H 7.98, N
^{3.76 1 H-NMR (DMSO-} d 6) δ (ppm): 1.2
0 (s, 18H, t-Bu), 3.40 (s, 3H, N
_{-CH 3), 3.56 (s,} 3H, -OCH 3), 7.0
8-7.80 (m, 6H, Ar-H) MS m / e: 397 (M ⁺ ), 247 (100) IR (KBr) ν (cm- ¹ ): 2980, 1720,
1620, 1590, 1370, 1220, 1110,
870,780

Production Example 33 0.91 g (5 mmol) of monomethyl terephthalate was added to 2 parts.
It was dissolved in 5 ml of tetrahydrofuran, 5 ml of thionyl chloride was added, and the mixture was refluxed for 3 hours while stirring. After cooling the reaction solution, tetrahydrofuran and thionyl chloride were distilled off under reduced pressure, the residue was dissolved in 25 ml of anhydrous ether, and 3,5-di-t-butyl-4-hydroxyaniline 1.
11 g (5 mmol) and 5 ml of anhydrous pyridine were added, and the mixture was stirred at room temperature for 3 hours. The same post-treatment as in Production Example 1 was performed below, and the residue was sequentially eluted with petroleum ether, dichloromethane, and ether by silica gel column chromatography to give 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carbamoyl. ] 1.3 g of methyl benzoate
Got mp 220-222 ° C

Production Example 34 According to the method of Production Example 29, 1.0 g (2.6 mmol) of the compound produced in Production Example 33 was hydrolyzed to give 4-.
0.50 g of [(3,5-di-t-butyl-4-hydroxyphenyl) carbamoyl] benzoic acid was obtained. mp 285-286 ° C (recrystallized from ethanol-petroleum ether)

Production Example 35 In accordance with the method of Production Example 31, 200 mg (0.54 mmol) of the compound produced in Production Example 34 was used to prepare 4- [N-methyl (3,5-di-t-butyl-4-methoxyphenyl) carbamoyl. ] 180 mg of methyl benzoate was obtained. mp 108-109 ° C MS m / e: 411 (M ⁺ , 69), 369 (M ⁺ -C).
H ₃ , 16), 163 (100)

Production Example 36 180 mg (0.44 mm) of the compound produced in Production Example 35
ol) was hydrolyzed in the same manner as in Production Example 2 to give 4- [N-methyl (3,5-di-t-butyl-4-methoxyphenyl).
80 mg of carbamoyl] benzoic acid was obtained.

Mp 218 to 219 ° C. (recrystallized from ethanol) Elemental analysis: C ₂₄ H ₃₁ NO ₄ = 397.50 Calculated value (%): C 72.51, H 7.86, N
3.54 Found (%): C 72.67, H 7.87, N
3.78 MS m / e: 397 (M ⁺ , 47), 382 (MC)
H ₃ , 27), 149 (100), 162 (30)

Production Example 37 4.5 g (18.3 mmol) of 3,5-di-t-butyl-4-hydroxyacetophenone and methyl 4-formylbenzoate were dissolved in 100 ml of absolute ethanol,
Add 6 g of dry barium hydroxide and stir 10
Heated to reflux for hours. After cooling the reaction solution, a solution prepared by dissolving 2 g of sodium hydroxide in 20 ml of water was added, and the mixture was heated under reflux for 2 hours while stirring. After cooling the reaction mixture, pH is adjusted to 5 with hydrochloric acid.
It was adjusted to ~ 6 and ethanol was distilled off under reduced pressure. 1 for residue
00 ml of water was added and the mixture was sufficiently stirred, the solid was collected by filtration, dried and crystallized with ethanol to give 4- [3-] as pale yellow crystals.
(3,5-di-t-butyl-4-hydroxyphenyl)
4.5 g of -3-oxo-1-propenyl] benzoic acid was obtained.

Mp 222-224 ° C. Elemental analysis: C ₂₄ H ₂₈ O ₄ = 380.46 Calculated value (%): C 75.75, H 7.42 Measured value (%): C 75.73, H 7. 38 ¹ H-NMR (DMSO-d ₆ ) δppm: 1.42
(S, 18H, t-Bu), 7.52-8.00 (m,
8H, Ar-H) MS m / e: 380 (M ⁺ , 30), 365 (M ⁺ -C).
H ₃ , 100), 233 (6) IR (KBr) ν (cm ^-1 ): 3620, 2960,
1780, 1755, 1605, 1570, 1420,
1320, 1290, 1210, 845, 780

Preparation Example 38 3,5-Di-t-butyl-4-methoxyacetophenone (3.0 g, 11 mmol) and methyl 4-formylbenzoate (1.88 g, 11 mmol) were added to 40 ml of anhydrous methanol and 5 ml of 20% sodium salt. The mixture was added to the methoxide solution, stirred at room temperature for 4 hours, and then left overnight. 10 ml of water was added to the reaction solution, and the mixture was stirred at room temperature overnight. PH with hydrochloric acid
Then, most of the methanol was distilled off under reduced pressure, the pH was adjusted to 2-3 with hydrochloric acid again, and the precipitated solid was collected by filtration and recrystallized with ethanol to give 4- [3- (3,5-di-). 3.8 g of t-butyl-4-methoxyphenyl) -3-oxo-1-propenyl] benzoic acid were obtained.

Mp 192-194 ° C. Elemental analysis: C ₂₅ H ₃₀ NO ₄ = 394.49 Calculated value (%): C 76.11, H 7.67 Measured value (%): C 75.75, H 7. 59 MS m / e: 394 (M ⁺ , 10), 378 (1
2), 261 (50), 247 (100), 149 (9
9)

Production Example 39 In the same manner as in Production Example 37, 0.65 g (2.62 mmo) of 3,5-di-t-butyl-4-hydroxyacetophenone
l) and 0.43 g of methyl 2-formylbenzoate
(2.62 mmol) to 2- [3- (3,5-di-t
-Butyl-4-hydroxyphenyl) -3-oxo-1
0.89 g of crude product of -propenyl] benzoic acid was obtained, which was crystallized with ethanol to obtain 0.72 g of yellow crystals.

Mp 105-106 ° C. Elemental analysis: C ₂₄ H ₂₈ O ₄ = 380.46 Calculated value (%): C 75.75, H 7.42 Measured value (%): C 75.60, H 6. 93 ¹ H-NMR (DMSO-d ₆ ) δppm: 1.40
(S, 18H, t-Bu), 5.38, 6.06 (m,
2H, CH = CH), 7.68 (m, 6H, Ar-H) MS m / e: 380 (M ⁺ , 67), 233 (10
0) IR (KBr) ν (cm ⁻¹ ): 3590, 2960,
2910, 1760, 1660, 1585, 1420,
1350, 1230, 880, 780, 745

Production Example 40 3,5-di-t-butyl-4-hydroxycinnamic acid 1.
38 g (5 mmol) was dissolved in 15 ml of anhydrous ether, 2 ml of oxalyl chloride was added, and the mixture was stirred at room temperature for 2 hours. The ether and excess oxalyl chloride were distilled off under reduced pressure, the residue was dissolved in 20 ml of anhydrous ether, 0.83 g (5 mmol) of ethyl 4-aminobenzoate and 5 ml of anhydrous pyridine were added, and the mixture was stirred at room temperature for 3 hours. After the reaction, ether was distilled off, and the residue was isopropyl ether-
Recrystallization from petroleum ether gave 1.6 g of ethyl 4- (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoate.

Mp 174 to 175 ° C. Elemental analysis: C ₂₆ H ₃₃ NO ₄ = 423.53 Calculated value (%): C 73.72, H 7.85, N
3.32 Found (%): C 74.23, H 7.60, N
_{^{3.30 1 H-NMR (CDCl 3}} ) δppm: 1.40 (s,
18H, t-Bu), 1.38 (t, 3H, -C)
_{H 3), 4.32 (q,} 2H, -O-CH 2 -), 6.2
9-7.84 (q, 2H, CH = CH), 7.30-
8.00 (m, 6H, Ar- H) MS m / e: 424 (M + + H, 94), 374 (M +
+ 1-OC ₂ H ₅ , 13), 259 (100) IR (KBr) ν (cm ⁻¹ ): 3610, 3300,
2960, 1710, 1660, 1620, 1590,
1520, 1425, 1380, 1270, 1170,
860,770

Production Example 41 In the same manner as in Production Example 40, 0.83 g (3 mmol) of 3,5-di-t-butyl-4-hydroxycinnamic acid and 4-
Aminobenzoic acid 0.41 g (3 mmol) to 4-
1.0 g of (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoic acid was obtained.

Mp 165 to 166 ° C. (crystal from ethanol) Elemental analysis: C ₂₄ H ₂₉ NO ₄ = 395.48 Calculated value (%): C 72.89, H 7.39, N
3.54 Found (%): C 72.81, H 7.50, N
^{3.83 1 H-NMR (DMSO-} d 6) δppm: 1.40
(S, 18H, t-Bu), 6.53, 7.50 (q,
2H, CH = CH), 7.36-7.91 (m, 6H,
Ar-H), 10.08 (s , 1H, -CO 2 H) MS m / e: 395 (M +, 13), 259 (10
0) IR (KBr) ν (cm ⁻¹ ): 3610, 3260,
2950, 1680, 1650, 1620, 1580,
1520, 1420, 1170, 850, 770

Production Example 42 In the same manner as in Production Example 40, 1.46 g (5.29 mmol) of 3,5-di-t-butyl-4-hydroxycinnamic acid and 0.80 g of methyl 2-aminobenzoate (5. 29 m
mol) to obtain 0.9 g of methyl 2- (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoate.

Mp 166 to 167 ° C. (crystal from ethanol) Elemental analysis: C ₂₅ H ₃₁ NO ₄ = 409.51 Calculated value (%): C 73.32, H 7.63, N
3.42 Found (%): C 72.75, H 7.67, N
3.37 MS m / e: 409 (M ⁺ , 30), 259 (10
0), 215 (10), 151 (12)

Production Example 43 1.0 g of methyl 2- (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoate (2.44 mm)
ol) was dissolved in 15 ml of ethanol, 3 ml of an aqueous solution containing 0.7 g of sodium hydroxide was added, and the mixture was left overnight with stirring at room temperature. The reaction solution was neutralized to pH 7 with hydrochloric acid, most of ethanol was distilled off under reduced pressure, and the pH was further adjusted with hydrochloric acid.
Adjusted to 3 and added 20 ml of water. The white solid thus formed was collected by filtration and crystallized from ethanol to obtain 0.72 g of 2- (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoic acid.

Mp 217 to 218 ° C. Elemental analysis: C ₂₄ H ₂₉ NO ₄ = 395.48 Calculated value (%): C 72.88, H 7.39, N
3.57 Found (%): C 72.86, H 7.25, N
3.64 MS m / e: 395 (M ⁺ , 20), 259 (10
0)

Production Example 44 0.3 g of powdery potassium hydroxide was mixed with 5 ml of dimethyl sulfoxide, and the mixture was stirred at room temperature for 5 minutes. 4-
270 mg (0.68 mmol) of (3,5-di-t-butyl-4-hydroxycinnamoylamino) benzoic acid and 0.7 ml of methyl iodide were added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was poured into 30 ml of water, saturated with sodium chloride, adjusted to pH 3 with hydrochloric acid, and adjusted to 3 with 20 ml of ether.
The extract was extracted twice, the ether layers were combined, washed with a saturated sodium chloride solution until neutral, and then washed with water. The ether layer was dried over magnesium sulfate, and the ether was distilled off under reduced pressure to obtain 2.0 g of a yellow solid. This was crystallized with ethanol, decolorized with activated carbon and treated with 4- [N-methyl- (3,5-di-t-
280 mg of methyl butyl-4-methoxycinnamoylamino) benzoate was obtained.

Mp 187-188 ° C. ¹ H-NMR (CDCl ₃ ) δppm: 1.38 (s,
18H, t-Bu), 3.41 (s, 3H, NC)
_{H 3), 3.62 (s,} 3H, O-CH 3), 3.96
(S, 3H, COOCH ₃ ), 6.10, 7.61
(Q, 2H, AB type, CH = CH), 7.19-8.1
0 (m, 6H, Ar-H) MS m / e: 438 (M ^{++ 1,} 16), 273
(100)

Production Example 45 In the same manner as in Production Example 43, 4- [N- was produced in Production Example 44.
Methyl methyl (3,5-di-t-butyl-4-methoxycinnamoylamino) benzoate 260 mg (0.59
mmol) to 4- [N-methyl- (3,5-di-t-
Butyl-4-methoxycinnamoylamino) benzoic acid 1
80 mg was obtained.

Mp 253-254 ° C. Elemental analysis: C ₂₆ H ₃₃ NO ₄ = 423.53 Calculated value (%): C 73.72, H 7.85, N
3.32 Found (%): C 73.46, H 7.75, N
^{3.43 1 H-NMR (DMSO-} d 6) δppm: 1.32
(S, 18H, t-Bu), 3.36 (s, 3H, N-
_{CH 3), 3.60 (s,} 3H, -OCH 3), 6.3
0, 7.50 (q, 2H, CH = CH), 7.22-
8.04 (m, 6H, Ar-H) MS m / e: 423 (M ⁺ , 7), 273 (10
0), 57 (t-Bu ⁺ , 54) IR (KBr) ν (cm ⁻¹ ): 2960, 1710,
1650, 1580, 1410, 1380, 1230,
1120, 850, 780

Production Example 46 4-methoxycarbonylcinnamic acid 1.52 g (7.37)
(mmol) was dissolved in 70 ml of anhydrous tetrahydrofuran, 10 ml of thionyl chloride was added, and the mixture was refluxed for 3 hours. Tetrahydrofuran and excess thionyl chloride were distilled off under reduced pressure, and the residue was dissolved in 70 ml of anhydrous ether.
5-di-t-butyl-4-hydroxyaniline 1.93
g (8.72 mmol) and 10 ml of anhydrous pyridine were added, and the mixture was stirred at room temperature for 3 hours. 50 ml of water was added to the reaction solution, the pH was adjusted to 3 with hydrochloric acid, the mixture was extracted 3 times with 30 ml of ether, the ether layers were combined, and washed with saturated saline and water. After drying over anhydrous magnesium sulfate, the ether was distilled off,
The residue was subjected to silica gel column chromatography, and eluted successively with petroleum ether, methylene chloride and ether,
The eluate was recrystallized from ethanol-petroleum ether to obtain 1.5 g of yellow crystals of 4- {2-[(3,5-di-t-butyl-4-hydroxyphenyl) carbamoyl) ethenyl} benzoate.

Mp 182 to 184 ° C. Elemental analysis: C ₂₅ H ₃₁ NO ₄ = 409.51 Calculated value (%): C 73.31, H 7.63, N
3.43 Found (%): C 73.19, H 7.82, N
3.40 ¹ H-NMR (CDCl ₃ ) δppm: 1.40 (s,
18H, t-Bu), 3.90 (s, 3H, -OC
H ₃ ), 6.50, 6.68 (q, 2H, CH = C
H), 7.40-8.04 (m, 6H, Ar-H) MS m / e: 409 (M ⁺ , 40), 221 (10).
0), 189 (14) IR (KBr) ν (cm ⁻¹ ): 3620,3380,
2980, 1710, 1670, 1600, 1550,
1440, 1370, 1280, 1180, 1110,
850, 780, 720

Production Example 48 In the same manner as in Production Example 43, 4- {2- produced in Production Example 47 was used.
Methyl [(3,5-di-t-butyl-4-hydroxyphenyl) carbamoyl) ethenyl} benzoate 0.8 g
0.3 g of 4- {2-[(3,5-di-t-butyl-4-hydroxyphenyl) carbamoyl) ethenyl} benzoic acid was obtained from (2 mmol).

Mp 328 to 332 ° C. Elemental analysis: C ₂₄ H ₂₉ NO ₄ = 395.49 Calculated value (%): C 72.89, H 7.39, N
3.54 Found (%): C 72.99, H 7.29, N
^{3.57 1 H-NMR (DMSO-} d 6) δppm: 1.40
(S, 18H, t-Bu), 6.70, 7.42 (q,
2H, CH = CH), 7.20-8.08 (m, 6H,
Ar-H) MS m / e: 396 (M ⁺ +1,25), 191 (8
0), 190 (100)

Example 1 4-[(3,5-di-t-butyl-4-hydroxyphenyl) carboxamide] Benzoic acid 1000 g, hydroxypropyl cellulose 800 g, crystalline cellulose 500
g, lactose 500 g, light anhydrous silicic acid 200 g and talc 200 g were formed into tablets having a diameter of 9 mm and a weight of 200 mg by a conventional method.

Example 2 100 g of 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzimidazole-6-carboxylic acid, 100 g of crystalline cellulose, 150 g of lactose and 20 g of light anhydrous silicic acid were encapsulated by a conventional method. I used it as an agent.

Example 3 Granules were produced by a conventional method according to the following formulation. (Formulation) 2- (3,5-di-t-butyl-4-hydroxyphenyl) benzoxazole-5-carboxylic acid 20
0 g, lactose 200 g, hydroxypropyl cellulose 2
0 g and talc 10 g were made into granules by a conventional method.

[Brief description of drawings]

FIG. 1 shows the expression state of oncogenes by autoradiography in Test Example 2.

FIG. 2 shows test results in Test Example 6.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI Technical display location A61K 31/19 8413-4C 31/235 8413-4C 31/275 8413-4C 31/415 9360-4C 31/42 9360-4C // C07D 235/12 257/00 7433-4C 263/56 (72) Inventor, Liu Hongyan, No. 1 Agricultural Research Center, Xuanwu District, Beijing, China (72) Inventor Kazuya Kamio 3-24-1 Takada, Toshima-ku, Tokyo Taisho Pharmaceutical Co., Ltd. (72) Inventor Shiro Nakaike 3-24-1, Takada, Toshima-ku, Tokyo Taisho Pharmaceutical Co., Ltd.

Claims (2)

[Claims] 1. A formula [Wherein, R ¹ represents a hydrogen atom, an alkyl group having 1 to 5 carbon atoms or an alkanoyl group having 2 to 5 carbon atoms, and X 1
Is vinylene group or the following formula (In the formula, R ² represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms.), R ³
Is an expression (In the formula, R ⁴ is a hydrogen atom, a halogen atom, a cyano group, a carbamoyl group, a sulfonamide group, a carboxyl group, an alkoxycarbonyl group having 2 to 5 carbon atoms, an alkyl group having 1 to 5 carbon atoms, or a carbon atom. A compound represented by the formulas 1 to 5 represents an alkoxy group, a trifluoromethyl group, an acetyl group or a tetrazolyl group, and R ⁵ represents a hydrogen atom, a hydroxyl group, a nitro group or an amino group, or a salt thereof as an active ingredient. Differentiation inducer. 2. A formula (In the formula, R ⁶ and R ⁷ are the same or different and each represents a hydrogen atom or an alkyl group having 1 to 5 carbon atoms, Y is a single bond or a vinylene group, and Z is O or NH.)
A differentiation-inducing agent containing, as an active ingredient, a fused heterocyclic carboxylic acid derivative represented by: