JPH11335375A - Benzamide derivative having histone deacetylase inhibiting action - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new benzamide derivative having strong histone deacetylase (HDA) inhibiting action. SOLUTION: The objective compound is expressed by formula I [A is H, a (substituted)phenyl or the like; X is direct bond, a group of formula (CH2 )e ((e) is 0-4) or the like; (n) is 0-4; Q is OCNR7 (R7 is H or the like) or the like; R1 is H, a halogen, amino or the like; Y is a group of formula II (Het is a heterocyclic group; R2 is H, a halogen or the like); R3 is amino or OH of a site adjacent to the benzamide bond bonding to the heterocyclic group], e.g. the compound of formula III. The compound of formula I can be produced by the condensation reaction of a compound of formula IV with a compound of formula V (E is amino group protected with protecting group used in the peptide-forming reaction such as t-butoxycarbonyl or nitro or the like) preferably at -20 to +50 deg.C for 0.5-48 hr and the removal of the protecting group to convert to amino group. The objective compound is useful as an agent for the treatment of diseases causing cell proliferation, an agent for increasing the effect of gene therapy, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a benzamide derivative having an inhibitory activity on histone deacetylase. More specifically, the present invention relates to the use of a histone deacetylase inhibitory activity for an anticancer drug and other drugs.

[0002]

2. Description of the Related Art In the nucleus of cells, DNA forms a complex with histones, maintains a highly folded chromatin structure, and is kept in an inactive state (Knezetic et al., Ce.
11, 45: 95-104, 1986). In order for the gene to be transcribed in the nucleus, it is necessary to bring the structure into an unfolded state so that various transcription factors can come into contact with DNA (Felsenfeld et al., Ce.
11, 86: 13-19, 1996). Although the relationship between histone acetylation and transcriptional activation has been reported for a long time, one of the actions that induces structural changes leading to transcriptional activation is one.
One was found to be histone acetylation (Hong et al., J. Biol. Chem., 268:
305-314, 1993, etc.). The acetylation is controlled by histone acetyltransferase.
e) and histone deacetylase (histone d)
eAcetylase; HDA), and its importance has been recently recognized (A. Corders, Bioch).
em. J. 265: 23, 1990). Sodium butyrate, which has long been confirmed to induce cell cycle arrest and differentiation, is a representative HDA inhibitor (LS Cou).
sens et al. Biol. Chem. , 254: 17
16, 1979), and clinical use was also attempted (N
ovogrodsky et al., Cancer, 51: 9-1.
4, 1983 and Miller et al., Eur. J. Ca
ncer Clin. Oncl. , 23: 1283-1.
287, 1987). However, since its basic inhibitory activity is low and its persistence in vivo is short, a high dose was required to show the effect. Therefore, a prodrug of butyric acid has been improved in sustainability (Zi-Xing et al., Can.
cer Res. , 54: 3494-3499, 199.
4 and Kasukabe et al., British J. et al. C
ancer, 75 (6): 850-854, 1997, etc.). In addition, trichostatin A (TSA), which is a natural product, stops cell cycle (Yoshida et al., Exp. Cell Res., 1).
77: 122-131, 1988), growth arrest, induction of differentiation (Yoshida et al., Cancer Res., 47: 368).
8-3691, 1987), which lead to changes in cell morphology and induction of apoptosis. As a mechanism, it has been confirmed that TSA is a highly active HDA inhibitor in vitro (Yoshida et al., J. Biol. C).
hem. 265: 17174, 1990). In addition, research on other HDA inhibitors has been continued, and trapoxins (Itazaki et al., J. Antibiot., 43
(12): 1524-1534, 1990), phenylbutyric acid (Carducci et al., Clin. Cance.
r Res. , 2 (2): 379, 1996, etc.). Since these HDA inhibitors have cell cycle arrest and differentiation inducing effects,
First, application to anticancer drugs is expected.

[0003] In addition, HDA inhibitors are expected to be applied to various other drugs. That is, as a therapeutic or ameliorating drug for diseases related to cell proliferation, for example, autoimmune diseases,
Skin diseases, infectious diseases (Darkin-Rattray et al., P
rc. Natl. Acad. Sci. USA, 93:
13143-13147, 1996), and the efficiency of vector introduction in gene therapy (Dion et al., Virology, 231: 201-2).
09, 1997), enhanced transgene expression (Chen
Proc. Natl. Acad. Sci. USA,
94: 5798-5803, 1997). However, none of the conventional inhibitors has reached a sufficiently satisfactory level as a pharmaceutical product in consideration of stability, toxicity, pharmacokinetics and active strength. Therefore, there is a strong demand for the development of drugs that solve these problems.

[0004]

SUMMARY OF THE INVENTION An object of the present invention is to provide a therapeutic and / or ameliorating agent for diseases related to cell proliferation and a drug for enhancing the effect of gene therapy, which have solved the problems of the conventional HDA inhibitors. It is to provide a compound useful as a pharmaceutical.

[0005]

Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that a novel benzamide derivative has a strong HDA inhibitory action, and have completed the present invention.

That is, the present invention relates to [1] a compound represented by the following formula (1):

[0007]

Embedded image [In the formula, A represents a hydrogen atom, a phenyl group which may be substituted, or a heterocyclic ring (as a substituent, a halogen atom, a hydroxyl group, an amino group, a nitro group, a cyano group, an alkyl group having 1 to 4 carbon atoms, 1-4 alkoxy groups, 1-4 carbon atoms
An aminoalkyl group, a C1-4 alkylamino group, a C1-4 acyl group, a C1-4 acylamino group, a C1-4 alkylthio group, a C1-4
Having 1 to 4 groups selected from the group consisting of a perfluoroalkyl group, a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group, an alkoxycarbonyl group having 1 to 4 carbon atoms, a phenyl group, and a heterocyclic ring. ). X is a direct bond or a compound of the formula (2)

[0008]

Embedded image 中 In the formula, e, g and m each independently represent an integer of 0-4. R4 is a hydrogen atom, an optionally substituted alkyl group having 1 to 4 carbon atoms or a group represented by the formula (3):

[0009]

Embedded image (In the formula, R6 represents an optionally substituted alkyl group having 1 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, a phenyl group or a heterocyclic ring). R5 represents any of the structures represented by｝ representing a hydrogen atom or an optionally substituted alkyl group having 1 to 4 carbon atoms. n represents the integer of 0-4. Q is the formula (4)

[0010]

Embedded image (Wherein, R 7 and R 8 each independently represent a hydrogen atom or an optionally substituted alkyl group having 1 to 4 carbon atoms). R1 represents a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, and a C1-4
Alkyl group, alkoxy group having 1 to 4 carbon atoms, 1 carbon atom
1 to 4 aminoalkyl groups, C1 to C4 alkylamino groups, C1 to C4 acyl groups, C1 to C4 acylamino groups, C1 to C4 alkylthio groups, C1 to C1
4 perfluoroalkyl group, C 1-4 perfluoroalkyloxy group, carboxyl group or 1 carbon atom
Represents an alkoxycarbonyl group of 1 to 4. Y is the equation (5)
[Formula 14]

[0011]

Embedded image (In the formula, Het represents a heterocyclic ring, and R2 represents a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, and an aminoalkyl having 1 to 4 carbon atoms. Group, an alkylamino group having 1 to 4 carbon atoms, an acyl group having 1 to 4 carbon atoms, an acylamino group having 1 to 4 carbon atoms,
Represents an alkylthio group having 1 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group or an alkoxycarbonyl group having 1 to 4 carbon atoms; To a possible position. R3 represents an amino group or a hydroxyl group existing at a position adjacent to the benzamide bond bonded to the heterocyclic ring. A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to claim 1 and a pharmaceutically acceptable salt represented by the formula:
[2] Formula (6)

[0012]

Embedded image [Wherein R9 is a halogen atom, a hydroxyl group, an amino group, a hydroxyamino group, an alkyl group having 1 to 4 carbon atoms,
4, an alkoxy group, an aminoalkyl group having 1 to 4 carbon atoms,
C1-C4 alkylamino group, C1-C4 acyl group, C1-C4 acylamino group, C1-C4 alkylthio group, C1-C4 perfluoroalkyl group, C-carbon Represents a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group, or an alkoxycarbonyl group having 1 to 4 carbon atoms. R1 and Y are as defined above. [3] a histone deacetylase inhibitor comprising as an active ingredient the benzamide derivative according to [1] and a pharmaceutically acceptable salt thereof, and [3] wherein Y is 4-aminothiophene-
A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to [2], which is 3-yl, and a pharmaceutically acceptable salt, and [4] a formula (7)

[0013]

Embedded image [Wherein, A, Y and R1 are as defined above. Z is given by the equation (8)
[Formula 17]

[0014]

Embedded image Means (Wherein, R7 and R8 have the same meanings as defined above.)], A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to [1] and a pharmaceutically acceptable salt. And [5] a histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to [4], wherein A is a pyridyl group which may be substituted, and a pharmaceutically acceptable salt, [6] A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to [5], wherein Y is 4-aminothiophen-3-yl, and a pharmaceutically acceptable salt. 7] Equation (9)
[Formula 18]

[0015]

Embedded image A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to [6] and a pharmaceutically acceptable salt thereof, and [8].
[7] An anticancer agent containing at least one of the compounds according to any one of [7] and [9].
It is a drug containing at least one of the compounds according to any one of claims [1] to [7] as an active ingredient.

[0016]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail.
The term "C1-4" as used herein refers to the number of carbon atoms per unit substituent. That is, for example, in the case of dialkyl substitution, it means having 2 to 8 carbon atoms.

The heterocyclic ring in the compound represented by the formula (1) means a nitrogen atom, an oxygen atom or a sulfur atom of 1 to 4
A monocyclic heterocyclic ring or a bicyclic fused heterocyclic ring comprising a 5-membered ring or a 6-membered ring, including, for example, pyridine, pyrazine, pyrimidine, pyridazine, thiophene, furan, pyrrole, pyrazole, Isoxazole, isothiazole, imidazole, oxazole, thiazole, piperidine, piperazine, pyrrolidine, quinuclidine, tetrahydrofuran, morpholine, thiomorpholine, etc. Condensed pyridine rings such as oxazolopyridine, imidazolopyridine, thiazolopyridine,
Benzofuran, benzothiophene, benzimidazole and the like can be mentioned.

The halogen atom includes a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

The alkyl group having 1 to 4 carbon atoms includes, for example, methyl group, ethyl group, n-propyl group, isopropyl group,
n-butyl group, isobutyl group, sec-butyl group, te
An rt-butyl group and the like can be mentioned.

The alkoxy group having 1 to 4 carbon atoms includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, allyloxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy. And the like. Examples of the aminoalkyl group having 1 to 4 carbon atoms include an aminomethyl group, a 1-aminoethyl group, and a 2-aminopropyl group.

The alkylamino group having 1 to 4 carbon atoms includes, for example, N-methylamino group, N, N-dimethylamino group,
N, N-diethylamino group, N-methyl-N-ethylamino group, N, N-diisopropylamino group and the like can be mentioned.

Examples of the acyl group having 1 to 4 carbon atoms include an acetyl group, a propanoyl group and a butanoyl group.

The acylamino group having 1 to 4 carbon atoms includes, for example, an acetylamino group, a propanoylamino group and a butanoylamino group.

The alkylthio group having 1 to 4 carbon atoms includes a methylthio group, an ethylthio group and a propylthio group.

The perfluoroalkyl group having 1 to 4 carbon atoms includes, for example, a trifluoromethyl group and a pentafluoroethyl group.

The perfluoroalkyloxy group having 1 to 4 carbon atoms includes, for example, a trifluoromethoxy group and a pentafluoroethoxy group.

Examples of the alkoxycarbonyl group having 1 to 4 carbon atoms include a methoxycarbonyl group and an ethoxycarbonyl group.

The optionally substituted alkyl group having 1 to 4 carbon atoms includes, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, s
ec-butyl group, tert-butyl group and the like, or a substituent thereof, having 1 to 4 groups selected from the group consisting of a halogen atom, a hydroxyl group, an amino group, a nitro group, a cyano group, a phenyl group and a heterocyclic ring. Things can be mentioned.

The salts of pharmaceutically acceptable compounds include inorganic acids commonly used in this field, such as hydrochloric acid, hydrobromic acid, sulfuric acid and phosphoric acid, acetic acid, lactic acid, tartaric acid, malic acid, succinic acid, and the like.
Examples thereof include salts with organic acids such as fumaric acid, maleic acid, citric acid, benzoic acid, trifluoroacetic acid, p-toluenesulfonic acid, and methanesulfonic acid.

Pharmaceuticals include anticancer drugs, skin diseases, infectious diseases,
When a compound represented by the formula (1) having an asymmetric carbon has different asymmetrical isomerism, it represents a therapeutic and / or ameliorating agent or an enhancer of a gene therapy effect for an allergic disease, an autoimmune disease, a vascular disease or the like. It can exist in the form or in the form of a mixture of stereoisomeric forms, including racemic forms. That is, the present invention also includes various forms defined as described above, and these can be similarly used as the active ingredient compound.

The typical structures of the compounds represented by the formula (7) of the present invention are specifically shown in Table 1 (Table 1 to Table 5). Note that the present invention is not limited to these examples.

[0032]

[Table 1]

[0033]

[Table 2]

[0034]

[Table 3]

[0035]

[Table 4]

[0036]

[Table 5] The compound of the present invention can be produced, for example, by the following method. Formula (10)

[0037]

Embedded image [Wherein, A, Q, X, R1 and n are as defined above. And a compound represented by the formula (11):

[0038]

Embedded image Wherein R2 and Het are as defined above. E is tert
An amino group or a nitro group protected with a protecting group used in ordinary peptide formation reactions such as a butoxycarbonyl group, a functional group which can be easily converted to an amino group such as an azido group, or a normal peptide formation reaction such as a benzyl group; Represents a hydroxyl group protected by a protecting group, which is attached to a position adjacent to an amino group. [12] which is obtained by subjecting a compound represented by the formula

[0039]

Embedded image (Where A, Q, X, R1, R2, E, Het and n
Is as defined above. The compound of the present invention can be obtained by removing the protecting group of the compound represented by the formula or converting the compound to an amino group.

The compound represented by the formula (10) and the compound represented by the formula (11)
The compound represented by is commercially available, or a known compound can be used. In the case of a novel compound, it can be produced by applying a synthesis method of a known compound. The condensation reaction of the formulas (10) and (11) can be carried out by an amide bond forming reaction in a usual peptide, for example, a method of an active ester or a mixed acid anhydride or an acid chloride. For example, a carboxylic acid component and 2, 4,
A phenol such as 5-trichlorophenol, pentachlorophenol or 4-nitrophenol, or an N-hydroxy compound such as N-hydroxysuccinimide or N-hydroxybenztriazole is condensed in the presence of dicyclohexylcarbodiimide to form an active compound. After conversion into an ester form, the reaction can be carried out by condensing with an amine component.

The carboxylic acid component may be oxalyl chloride,
The reaction can be carried out by reacting with thionyl chloride, phosphorus oxychloride, or the like, converting the acid chloride into an acid chloride, and then condensing with an amine component. Alternatively, the reaction can be performed by reacting a carboxylic acid component with isobutyl chlorocarbonate or methanesulfonyl chloride to obtain a mixed acid anhydride, and then condensing the mixed acid anhydride with an amine component.

Further, the condensation reaction is carried out by peptide condensation of dicyclohexylcarbodiimide, N, N'-carbonyldiimidazole, diphenylphosphate azide, diethylphosphate cyanide, 2-chloro-1,3-dimethylimidazolonium chloride, etc. It can also be performed using the reagent alone.

The reaction is usually carried out at -20 to + 50 ° C. for 0.5 to
Perform for 48 hours. Examples of the solvent used include aromatic hydrocarbons such as benzene and toluene, ethers such as tetrahydrofuran, dioxane and diethyl ether, halogenated hydrocarbons such as methylene chloride and chloroform, and N, N-dimethylformamide. Examples thereof include alcohols such as methanol and ethanol, and mixtures thereof. If necessary, an organic base such as triethylamine or pyridine is added to react.

The benzamide derivative having an inhibitory effect on histone deacetylase of the present invention is useful as an agent for treating and / or ameliorating a disease relating to cell proliferation, an agent for enhancing the effect of gene therapy or an immunosuppressant.

Here, the diseases related to cell proliferation include malignant tumors, autoimmune diseases, skin diseases, infectious diseases, vascular diseases,
Allergic diseases, gastrointestinal tract injury, hormonal diseases, diabetes and the like.

Malignant tumors include hematopoietic tumors such as acute leukemia, chronic leukemia, malignant lymphoma, multiple myeloma, and macroglobulinemia, as well as colorectal cancer, brain tumor, head and neck cancer, breast cancer,
Lung, esophagus, gastric, liver, gallbladder, bile duct, pancreatic, islet cell, renal cell, adrenal cortical, bladder, prostate,
Solid tumors such as testicular tumor, ovarian cancer, uterine cancer, choriocarcinoma, thyroid cancer, malignant carcinoid tumor, skin cancer, malignant melanoma, osteosarcoma, soft tissue sarcoma, neuroblastoma, Wilms tumor, retinoblastoma No.

Autoimmune diseases include rheumatism, nephritis, diabetes, systemic lupus erythematosus, human autoimmune lymphoproliferative lymphadenopathy, immunoblastic lymphadenopathy, Crohn's disease, ulcerative colitis and the like. .

The skin diseases include psoriasis, acne, eczema, atopic dermatitis, parasitic skin diseases, alopecia, purulent skin diseases, skin sclerosis and the like.

[0049] Infectious disease means a disease caused by infection of various bacteria, viruses or parasites.

The vascular disease includes a drug for treating arteriosclerosis or restenosis.

The enhancement of the effect of the gene therapy includes, for example, increasing the efficiency of introduction of a gene vector and enhancing the expression of a transgene. The target disease of the present invention is not limited to these.

The active ingredient compounds of the present invention are useful as pharmaceuticals, and they are used in the form of general medical preparations. The formulation contains commonly used fillers, extenders, binders,
It is prepared using diluents or excipients such as humectants, disintegrants, surfactants and lubricants. Various forms of this pharmaceutical preparation can be selected according to the purpose of treatment, and typical examples are tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, injections (solutions, suspensions). And suppositories.

In molding into tablets, various carriers well known in the art can be widely used. Examples include lactose, glucose, starch, calcium carbonate, kaolin,
Excipients such as crystalline cellulose and silicic acid, water, ethanol,
Binders such as propyl alcohol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, polyvinylpyrrolidone, disintegrators such as dried starch, sodium alginate, agar powder, carmellose calcium, starch, lactose, etc. Disintegration inhibitors such as sucrose, cocoa butter, hydrogenated oils, etc., absorption promoters such as quaternary ammonium bases, sodium lauryl sulfate, humectants such as glycerin and starch, starch, lactose, kaolin, bentonite, colloidal silicic acid, etc. And a lubricant such as talc, stearate and polyethylene glycol. Furthermore, tablets can be made into tablets coated with a usual coating, if necessary, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets or two-layer tablets or multilayer tablets.

For molding into a pill form, a wide variety of carriers conventionally known in this field can be used. Examples include crystalline cellulose, lactose, starch,
Excipients such as hardened vegetable oil, kaolin, talc, etc., binders such as gum arabic powder, tragacanth powder, gelatin and the like, disintegrants such as carmellose calcium, agar and the like can be mentioned.

Capsules are prepared by mixing the active ingredient compound with the various carriers exemplified above and filling the resulting mixture into hard gelatin capsules, soft capsules, and the like, according to a conventional method.

When prepared as an injection, the liquids, emulsions and suspensions are preferably sterilized and isotonic with blood. In shaping these forms, they are commonly used as diluents in this field. Things, such as water,
Ethanol, macrogol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like can be used. In this case, an amount of salt, glucose or glycerin necessary for preparing an isotonic solution may be contained in the pharmaceutical preparation,
Further, ordinary solubilizers, buffers, soothing agents and the like may be added.

For shaping in the form of suppositories, conventionally known carriers can be widely used. Examples thereof include semi-synthetic glycerides, cocoa butter, higher alcohols, esters of higher alcohols, polyethylene glycol and the like.

Further, if necessary, a coloring agent, a preservative, a flavor, a flavoring agent, a sweetening agent, and other pharmaceuticals can be contained in the pharmaceutical preparation.

The amount of the active ingredient compound to be contained in these pharmaceutical preparations of the present invention is not particularly limited and may be appropriately selected from a wide range, but is usually about 1 to 70% by weight, preferably about 1 to 70% by weight in the preparation composition. Is preferably about 5 to 50% by weight.

The method of administering these pharmaceutical preparations of the present invention is not particularly limited, and the pharmaceutical preparations are administered according to various preparation forms, the age, sex, degree of disease and other conditions of the patient. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules are administered orally, and injections are
It is administered intravenously, alone or as a mixture with a normal replenisher such as glucose or amino acids, and if necessary, intramuscularly, subcutaneously or intraperitoneally. Suppositories are administered rectally.

The dose of these pharmaceutical preparations of the present invention is appropriately selected depending on the usage, age, sex, degree of disease and other conditions of the patient. Usually, the amount of the active ingredient compound is one per kg of body weight. It is preferable to use about 0.0001 to 100 mg per day. It is desirable that the active ingredient compound be contained in a dosage unit form in a range of about 0.001 to 1,000 mg.

The compounds of the present invention represented by the formula (1) and salts thereof do not show toxicity at doses that are pharmacologically effective.

[0063]

The present invention will be described below in detail with reference to examples.
The present invention is not limited to these. Example 1 Synthesis of N- (4-aminothiophen-3-yl) -4-methoxybenzamide (1-1) 2,5-dibromo-
3.0 g (9 mmol) of 3,4-dinitrothiophene was added to 60 ml of concentrated hydrochloric acid, and 6.5 g (54 mmol) of tin was added. After hydrogen evolution ceased, it was left at room temperature overnight. The precipitated solid was separated by filtration, and the filtrate was concentrated to about 20 ml. The crystals precipitated by ice cooling are collected by filtration, and the obtained crystals are washed with ether, 3,4-diaminothiophene tin chloride,
0.39 g (10%) of the dihydrochloride was obtained. 1H NMR (270 MHz, DMSO-d6) δ ppm: 5.9 (br.s) (1-2) 3,4-diaminothiophene tin chloride, 2
0.39 g (0.87 mmol) of hydrochloride and 74 mg (0.43 mmol) of p-anisoyl chloride were suspended in chloroform, and 1 ml of triethylamine was added under ice cooling.
It returned to room temperature and left overnight. Dip into 0.3N hydrochloric acid CHCl
Extracted in ₃ . The aqueous layer was neutralized with NaOH, and the crystals were collected by filtration. The crystals were dissolved in CHCl ₃ / MeOH and dried over MgSO ₄ . Then, the mixture was filtered and concentrated to obtain 40 mg (19%) of the target N- (4-aminothiophen-3-yl) -4-methoxybenzamide as crystals. mp 126 ° C (dec.) 1H NMR (270MHz, DMSO-d6) δppm: 4.9 (2H, br.s), 6.13
(1H, d, J = 3.7Hz), 7.05 (2H, d, J = 8.8Hz), 7.46 (1H,
t, J = 3.7Hz), 7.93 (2H, d, J = 8Hz), 9.53 (1H, s)

Example 2 4-amino-N- (4-aminothiophen-3-yl)
Synthesis of benzamide (2-1) 3,4-diaminothiophene tin chloride, 2
2.33 g (7 mmol) of hydrochloride and 648 mg (3.5 mmol) of p-nitrobenzoyl chloride were suspended in chloroform, and 5 ml of triethylamine was added under ice cooling.
It returned to room temperature and left overnight. The mixture was poured into diluted sulfuric acid and extracted with CHCl ₃ . The aqueous layer was neutralized with NaOH, and the crystals were collected by filtration. The crystals are dissolved in CHCl ₃ / MeOH and purified on a short column to give the desired N- (4-aminothiophen-3-yl) -4
40 mg (5%) of -nitrobenzamide as crystals were obtained. 1H NMR (270MHz, DMSO-d6) δppm: 4.9 (2H, br.s), 6.15
(1H, d, J = 3.7Hz), 7.54 (1H, d, J = 3.7Hz), 8.15 (2H,
d, J = 8Hz), 8.37 (2H, d, J = 8Hz), 9.97 (1H, s) (2-2) N- (4-aminothiophen-3-yl)-
0.186 g of 4-nitrobenzamide (0.71 mmol
l) was suspended in 1 ml of methanol and 1 ml of concentrated hydrochloric acid, 402 mg (2.1 mmol) of tin dichloride was added, and the mixture was stirred at room temperature and left overnight. Ice was added, neutralized with 3N NaOH, and extracted with CHCl ₃ . The organic layer and the solid were purified by a silica gel column, and 35 mg of the desired 4-amino-N- (4-aminothiophen-3-yl) benzamide was used as crystals.
(21%). mp 130 ° C (dec.) 1H NMR (270MHz, DMSO-d6) δppm: 4.8 (2H, br.s), 5.7 (2
H, br.s), 6.11 (1H, d, J = 3.7Hz), 6.59 (2H, d, J = 8H
z), 7.41 (1H, d, J = 3.7Hz), 7.68 (2H, d, J = 8Hz), 9.2
(1H, s) IR (KBr) cm-1; 3387,1603,1506,1405,1286,1185,845,772

Example 3 Synthesis of N- (4-aminothiophen-3-yl) -4-hydroxyaminobenzamide 0.04 g (0.152 mmol) of N- (4-aminothiophen-3-yl) -4-nitrobenzamide ) Was dissolved in 5 ml of methanol and hydrogenated with Pd / C. The catalyst was filtered off and the filtrate was concentrated to obtain crystals. After washing with chloroform and drying, 20 mg (53%) of the desired N- (4-aminothiophen-3-yl) -4-hydroxyaminobenzamide was obtained. mp150 ° C (dec.) 1H NMR (270MHz, DMSO-d6) δppm: 4.85 (2H, br.s), 6.1
1 (1H, d, J = 3.7Hz), 6.87 (2H, d, J = 8Hz), 7.44 (1H, d,
J = 3.7Hz), 7.80 (2H, d, J = 8Hz), 8.57 (1H, s), 8.78 (1H,
s), 9.4 (1H, s) IR (KBr) cm-1; 3281,1605,1507,1286,1182,847,767

Example 4 Synthesis of N- (3-aminopyridin-2-yl) -4-aminobenzamide (4-1) 1.33 g of 2-amino-3-nitropyridine
4-nitrobenzoyl chloride in dioxane solution of 1.
86 g was added and the mixture was heated to reflux. Then, dioxane was distilled off, and the precipitated crystal was recrystallized from ethyl acetate / methanol to give N-
(1.74 g (60%) of 3-nitro-2-pyridyl-4-nitrobenzamide was obtained.) Mp215-216 ° C. 1H NMR (270 MHz, DMSO-d6) δ ppm: 7.58 (1H, dd, J = 5, 8H)
z), 8.23 (2H, d, J = 9Hz), 8.38 (2H, d, J = 9Hz), 8.48 (1H, dd, J = 1.5, 8Hz), 8.79
(1H, dd, J = 1.5, 5Hz) (4-2) N- (3-nitropyridin-2-yl) -4
1.0 g of nitrobenzamide was suspended in a mixed solvent of 4 ml of DMF and 12 ml of methanol, and hydrogenation was performed using 10% palladium carbon as a catalyst. After filtering off the catalyst, the solvent was concentrated and crystallized from methanol to give N- (3-aminopyridin-2-yl) -4-aminobenzamide in 0.6 ml.
9 g (76%) were obtained. mp162-163 ° C 1H NMR (270MHz, DMSO-d6) δppm: 4.92 (2H, br.s), 5.75
(2H, s), 6.58 (2H, d, J = 9Hz), 7.04 (1H, dd, J = 5, 8H
z), 7.17 (1H, d, J = 6.6Hz), 7.73 (1H, d, J = 5Hz), 7.77
(2H, d, J = 9Hz), 9.96 (1H, s)

Example 5 N- (4-aminothiophen-3-yl) -4- {N-
Synthesis of (pyridin-3-yl-methoxycarbonyl) aminomethyl {benzamide 4- (N- (pyridin-3-yl-methoxycarbonyl) aminomethyl} benzoic acid (208 mg) was suspended in methylene chloride (6.2 ml) and cooled under ice-cooling. Oxalyl chloride 0.
32 ml were added. After stirring at room temperature for 30 minutes, the mixture was dried under reduced pressure. The obtained 4- {N- (pyridin-3-yl-methoxycarbonyl) aminomethyl} benzoyl chloride was condensed in the same manner as in Example 2 to give N- (4-aminothiophen-3-yl) -4-. {N- (pyridin-3-yl-methoxycarbonyl) aminomethyl} benzamide was obtained in an amount of 55 mg (20%). mp. 170-174 ° C. 1H NMR (270MHz, DMSO-d6) δppm: 4.33 (2H, d, J = 5.9H)
z), 4.91 (2H, br.s), 5.15 (2H, s), 6.18 (1H, d, J = 3.
7), 7.44 (2H, d, J = 8.1), 7.55 (1H, d, J = 2.7), 7.83 (1
H, d, J = 8.1) 7.94 (2H, d, J = 8.1), 8.01 (1H, m), 8.58
(1H, d, J = 3.7), 8.64 (1H, s), 9.67 (1H, s) .IR (KBr) cm-1: 3222, 3029, 1713, 1651, 1555, 1481, 1
419, 1266, 1129, 1024,982, 856, 793, 708.

Example 6 N- (4-aminothiophen-3-yl) -4- {N-
Synthesis of (2-nitrobenzenesulfonyl) amino {benzamide 4- {N- (2-nitrobenzenesulfonyl) amino}
The reaction was carried out in the same manner as in Example 5 using benzoic acid, and N- (4-
Aminothiophen-3-yl) -4- {N- (2-nitrobenzenesulfonyl) amino} benzamide was obtained. 1H NMR (270MHz, DMSO-d6) δppm: 6.10 (1H, d), 7.22 (2
(H, d, J = 8Hz), 7.45 (1H, d), 7.8-7.9 (4H, m), 8.0-8.1
(2H, m), 9.54 (1H, s) IR (KBr) cm-1: 3100, 1609, 1541, 1507, 1347, 1164, 8
53, 778

Pharmacological Test Example 1 (Histone Deacetylase)
Inhibitory action) (1) [^ThreeH] Preparation of acetyl histone K562 cells (10⁸) To [^ThreeH] Sodium n-butyrate
And the method of Yoshida et al. (J. Biol. Chem.,
265: 17174, 1990)
Issued. (2) Partial purification of histone deacetylase K562 cells (2.5 × 10⁸Nucleus collected from
Tada et al.'S method (J. Biol. Chem., 265: 17).
174, 1990).
Using oQ HR5 / 5 (Pharmacia), 0-1M
Histone deacetylase by NaCl concentration gradient
Was partially purified. (3) Measurement of histone deacetylase inhibitory activity Prepared in (1) [^ThreeH] acetyl histone 100μ
g / ml and histone deacetylase prepared in (2)
Buffer A containing 2 μl of fraction [Composition: 5 mM potassium phosphate
(PH 7.5), 5% glycerol, 13 mM ED
TA] in 50 μl for 10 minutes at 37 ° C.
Was. After stopping the reaction by adding 2.5N hydrochloric acid, acetic acid was added.
Add 550 μl of ethyl, vortex and centrifuge.
Add 400 μl of ethyl acetate layer to scintillation via
And add 2 ml of scintillator to allow reaction.
Released^Three[H] acetic acid radioactivity was measured. Histo
For the measurement of deacetylase inhibitory activity, the test compound
After dissolving in MSO, appropriately dilute with buffer A and add to the reaction system
The concentration of the drug that causes 50% enzyme inhibition (I
C ₅₀: ΜM). The experimental results are shown in Table 2 below.
The results are shown in [Table 6].

[0070]

[Table 6]

[0071]

The benzamide derivative of the present invention having an inhibitory effect on histone deacetylase is useful as a therapeutic and / or ameliorating agent for diseases related to cell proliferation, an effect-enhancing agent for gene therapy or an immunosuppressant. In particular, it is highly effective as an anticancer agent and is effective against hematopoietic tumors and solid cancers.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Osamu Nakanishi 1900-1, Togo, Mobara City, Chiba Prefecture Inside Mitsui Pharmaceutical Industry Co., Ltd. (72) Inventor Yukiyasu Mariko 1900-1 Togo, Mobara-shi, Chiba Mitsui Pharmaceutical Co., Ltd.

Claims (9)

[Claims] 1. A compound represented by the formula (1): [In the formula, A represents a hydrogen atom, a phenyl group which may be substituted, or a heterocyclic ring (as a substituent, a halogen atom, a hydroxyl group, an amino group, a nitro group, a cyano group, an alkyl group having 1 to 4 carbon atoms, 1-4 alkoxy groups, 1-4 carbon atoms
An aminoalkyl group, a C1-4 alkylamino group, a C1-4 acyl group, a C1-4 acylamino group, a C1-4 alkylthio group, a C1-4
Having 1 to 4 groups selected from the group consisting of a perfluoroalkyl group, a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group, an alkoxycarbonyl group having 1 to 4 carbon atoms, a phenyl group, and a heterocyclic ring. ). X is a direct bond or a compound of the formula (2) 中 In the formula, e, g and m each independently represent an integer of 0-4. R4 is a hydrogen atom, an optionally substituted alkyl group having 1 to 4 carbon atoms or a group represented by the formula (3): (In the formula, R6 represents an optionally substituted alkyl group having 1 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, a phenyl group or a heterocyclic ring). R5 represents any of the structures represented by｝ representing a hydrogen atom or an optionally substituted alkyl group having 1 to 4 carbon atoms. n represents the integer of 0-4. Q is the formula (4) [Formula 4] (Wherein, R 7 and R 8 each independently represent a hydrogen atom or an optionally substituted alkyl group having 1 to 4 carbon atoms). R1 represents a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, and a C1-4
Alkyl group, alkoxy group having 1 to 4 carbon atoms, 1 carbon atom
1 to 4 aminoalkyl groups, C1 to C4 alkylamino groups, C1 to C4 acyl groups, C1 to C4 acylamino groups, C1 to C4 alkylthio groups, C1 to C1
4 perfluoroalkyl group, C 1-4 perfluoroalkyloxy group, carboxyl group or 1 carbon atom
Represents an alkoxycarbonyl group of 1 to 4. Y is the equation (5)
[Chemical Formula 5] (In the formula, Het represents a heterocyclic ring, and R2 represents a hydrogen atom, a halogen atom, a hydroxyl group, an amino group, an alkyl group having 1 to 4 carbon atoms, an alkoxy group having 1 to 4 carbon atoms, and an aminoalkyl having 1 to 4 carbon atoms. Group, an alkylamino group having 1 to 4 carbon atoms, an acyl group having 1 to 4 carbon atoms, an acylamino group having 1 to 4 carbon atoms,
Represents an alkylthio group having 1 to 4 carbon atoms, a perfluoroalkyl group having 1 to 4 carbon atoms, a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group or an alkoxycarbonyl group having 1 to 4 carbon atoms; To a possible position. R3 represents an amino group or a hydroxyl group existing at a position adjacent to the benzamide bond bonded to the heterocyclic ring. And a pharmaceutically acceptable salt thereof as an active ingredient. 2. The compound of the formula (6) [Wherein R9 is a halogen atom, a hydroxyl group, an amino group, a hydroxyamino group, an alkyl group having 1 to 4 carbon atoms,
4, an alkoxy group, an aminoalkyl group having 1 to 4 carbon atoms,
C1-C4 alkylamino group, C1-C4 acyl group, C1-C4 acylamino group, C1-C4 alkylthio group, C1-C4 perfluoroalkyl group, C-carbon Represents a perfluoroalkyloxy group having 1 to 4 carbon atoms, a carboxyl group, or an alkoxycarbonyl group having 1 to 4 carbon atoms. R1 and Y are as defined above. ] A histone deacetylase inhibitor comprising the benzamide derivative according to claim 1 and a pharmaceutically acceptable salt as an active ingredient. 3. A histone deacetylase inhibitor comprising, as an active ingredient, the benzamide derivative according to claim 2, wherein Y is 4-aminothiophen-3-yl. 4. A compound of the formula (7) [Wherein, A, Y and R1 are as defined above. Z is given by the equation (8)
[Formula 8] [Formula 8] Means (Wherein R7 and R8 have the same meanings as defined above.)], Wherein the benzamide derivative according to claim 1 and a pharmaceutically acceptable salt thereof are a histone deacetylase inhibitor comprising an active ingredient. . 5. The histone deacetylase inhibitor comprising as an active ingredient the benzamide derivative according to claim 4, wherein A is a pyridyl group which may be substituted, and a pharmaceutically acceptable salt. 6. A histone deacetylase inhibitor comprising as an active ingredient a benzamide derivative according to claim 5, wherein Y is 4-aminothiophen-3-yl, and a pharmaceutically acceptable salt. 7. A compound represented by the formula (9): A histone deacetylase inhibitor comprising, as an active ingredient, a benzamide derivative represented by the formula and a pharmaceutically acceptable salt. 8. An anticancer agent comprising at least one of the compounds according to claim 1 as an active ingredient. 9. A pharmaceutical comprising at least one of the compounds according to claim 1 as an active ingredient.