JP2009191041A - Hydroxymethyl ketone derivative - Google Patents

Abstract

[式中、Ｒは、 無置換又は置換基を有していても良いフェニル基、無置換または置換基を有していても良いベンジル基、無置換または置換基を有していても良いフェネチル基を表し、ＸはＯ、ＣＨ_２又はＳを表し、ＹはＣＨ_２を表し、Ｘ−Ｙは二重結合又は三重結合でも良く、ＺはＮ又はＣＨを表し、Ｒ’は水素原子、低級アルキル基を表す］で表されるヒドロキシメチルケトン誘導体若しくはその薬理学上許容される塩又はそれらの水和物。
【選択図】図１The present invention provides a novel compound that selectively inhibits the activity of histone deacetylase 6 (HDAC6), and a medicament containing the compound.
The following general formula (1)

[Wherein, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. Represents a group, X represents O, CH ₂ or S, Y represents CH ₂ , XY may be a double bond or a triple bond, Z represents N or CH, R ′ represents a hydrogen atom, lower A hydroxymethyl ketone derivative represented by the formula: or a pharmacologically acceptable salt thereof, or a hydrate thereof.
[Selection] Figure 1

Description

  The present invention relates to a hydroxymethyl ketone derivative effective as a selective inhibitor of histone deacetylase 6 (hereinafter also referred to as HDAC6), an addition salt thereof or a hydrate thereof, and a pharmaceutical containing these compounds.

  It is known that the chromatin structure constructed with DNA and histone as main components plays an important role in the transcription and control of genes in the nucleus of eukaryotes. The basic structural unit of chromatin is a structure in which DNA of 146 base pairs is composed of approximately two turns of histone octamers (consisting of two molecules each of histones H2A, H2B, H3, and H4), and is called a nucleosome ( Non-patent document 1). In a normal nucleosome structure, gene transcription is in a repressed state, and the nucleosome structure needs to be relaxed in order to be activated. One of the important mechanisms for controlling transcriptional activity is histone acetylation and deacetylation (Non-patent Document 2).

  Histone acetylation and deacetylation are performed by histone acetyltransferase (histone acetylase, HAT) and histone deacetylase (histone deacetylase, hereinafter also referred to as HDAC), respectively. So far, 18 families of HDACs have been identified, and are further classified into zinc-dependent groups containing zinc at the active center (HDAC1-11) and NAD + -dependent groups (SIRT1-7) (non- Patent Document 3). Each isoform included in the zinc-dependent group forms and functions as a large complex by interacting with many other proteins. For example, HDAC1 and HDAC2 are present in the SIN3 complex and NURD / Mi2 complex, and HDAC3 forms a complex with NCoR and SMRT and is known to be involved in transcriptional repression of retinoid receptors and thyroid receptors. It has been.

  Among the groups containing zinc at the active center, HDAC6 is localized in the cytoplasm, unlike other HDACs, and deacetylates α-tubulin, non-histone proteins such as HSP90 and contactin to stabilize microtubules. It is unique in that it is suggested to be involved in molecular chaperone activity control and cell movement (Non-patent Document 3). And the inhibitor of HDAC6 causes the growth inhibition of myeloma cells without affecting non-cancerous cells (Non-patent Document 4), etc., suggesting the usefulness of HDAC6 inhibitors as anticancer agents or anticancer agents. Yes. Furthermore, inhibition of HDAC6 activity to promote tubulin acetylation improves the reduction in microtubule-dependent intracellular transport characteristic of Huntington's disease and the like (Non-patent Document 5). It is also expected as a therapeutic drug.

So far, many drugs such as butyric acid, trichostatin A, SAHA, MS-275 have been known as HDAC inhibitors, and cell cycle arrest, morphology normalization / differentiation inducing action of transformed cells (non-patent literature) 6) It is suggested that it is useful as an anticancer agent or anticancer agent from the effects of reducing the cell cycle, inducing differentiation and inducing apoptosis (Non-patent Documents 7 and 8) (Non-patent Document 10).
In recent years, it has become clear that each isoform belonging to the HDAC family may have a unique role. Among them, HDAC6 is localized in the cytoplasm and plays a different function from other isoforms as described above. It has been reported.
Therefore, the development of an inhibitor that selectively acts on each isoform is strongly desired in terms of reducing side effects, even if it is a drug that exhibits an inhibitory action on HDAC, rather than a conventional highly versatile drug. ing. So far, several drugs that selectively act on HDAC6 have been reported (Non-patent Document 3), but further development of drugs that exhibit superior effects in terms of selectivity, activity, and safety is required. Yes.

Wolfe et al., Cell, 84: 817-819, 1996. Hebbes et al., EMBO J, 7: 1395-1402, 1988 Itoh et al. Med. Chem. , 50: 5425-5438, 2007. Hideshima et al., Proc. Natl. Acad. Sci. U. S. A. , 102: 8567-8572, 2005. Dompierre et al., J. MoI. Neurosci. , 27: 3571-3583, 2007 Counsens et al. Biol. Chem., 254: 1716-1723, 1979. Yoshida et al., Cancer Res., 47: 3688-3691, 1987 Yoshida et al., Exp. Cell Res., 177: 122-131, 1988. Yoshida et al. Biol. Chem. , 265: 17174-17179, 1990. Marks et al. Natl. Cancer Inst. , 92: 1210-1216, 2000

The present invention selectively inhibits the activity of HDAC6 (compared to HDAC1 to 5), provides a novel compound having low toxicity to living bodies, a method for producing the same, and an HDAC6 inhibitor containing the novel compound as an active ingredient For the purpose of provision.
Another object of the present invention is to provide a medicament comprising the novel compound as an active ingredient and effective for treating a disease associated with enhanced HDAC6 activity.
Furthermore, an object of the present invention is to provide a method for treating a disease associated with enhanced HDAC6 activity using the above-mentioned medicament.

As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that a novel hydroxymethyl ketone derivative represented by the following general formula (1) has an HDAC6-selective inhibitory action. It came to complete.
So far, compounds having N-hydroxycarbamoyl group, thiol group, aminobenzamide group or axylanyl group are known as compounds having HDAC inhibitory activity, but any compound is stable and toxic in vivo. There is also concern about the possibility of causing side effects.
The compound in the present invention is a compound having an α-hydroxyketone at the zinc binding site in order to overcome the problems of existing HDAC inhibitors, and exhibits a selective inhibitory effect on HDAC6.

[式中、Ｒは、 無置換又は置換基を有していても良いフェニル基、無置換または置換基を有していても良いベンジル基、無置換または置換基を有していても良いフェネチル基を表し、ＸはＯ、ＣＨ_２又はＳを表し、ＹはＣＨ_２を表し、Ｘ−Ｙは二重結合又は三重結合でも良く、ＺはＮ又はＣＨを表し、Ｒ’は水素原子、低級アルキル基を表す］で表されるヒドロキシメチルケトン誘導体若しくはその薬理学上許容される塩又はそれらの水和物。 That is, the present invention
General formula (1)

[Wherein, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. Represents a group, X represents O, CH ₂ or S, Y represents CH ₂ , XY may be a double bond or a triple bond, Z represents N or CH, R ′ represents a hydrogen atom, lower A hydroxymethyl ketone derivative represented by the formula: or a pharmacologically acceptable salt thereof, or a hydrate thereof.

[式中、Ｒは、 無置換又は置換基を有していても良いフェニル基、無置換または置換基を有していても良いベンジル基、無置換または置換基を有していても良いフェネチル基を表し、ＸはＯ、ＣＨ_２又はＳを表し、ＹはＣＨ_２を表し、Ｘ−Ｙは二重結合又は三重結合でも良く、ＺはＮ又はＣＨを表し、Ｒ’は水素原子、低級アルキル基を表す］で表されるヒドロキシメチルケトン誘導体若しくはその薬理学上許容される塩又はそれらの水和物、並びに薬理学上許容される担体を含有するＨＤＡＣ６阻害剤又はＨＤＡＣ６活性の亢進に起因する疾患の治療のための医薬。 General formula (1)

[Wherein, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. Represents a group, X represents O, CH ₂ or S, Y represents CH ₂ , XY may be a double bond or a triple bond, Z represents N or CH, R ′ represents a hydrogen atom, lower Resulting from the enhancement of HDAC6 inhibitor or HDAC6 activity containing a hydroxymethyl ketone derivative represented by the above formula or a pharmacologically acceptable salt thereof or a hydrate thereof, and a pharmacologically acceptable carrier Medicine for the treatment of diseases.

  The compounds of the present invention are structurally novel and selectively inhibit the activity of HDAC6 among HDAC isoforms. As a result, it is useful for the development of a drug that selectively shows an effect on a disease caused by the enhanced HDAC6 activity.

  Since the compound of the present invention has high selectivity for HDAC6, it can be expected to have higher safety when taken for a long time than other HDAC-inhibiting compounds showing general-purpose effects.

  Since the compound of the present invention synergistically promotes the effect of an anticancer agent or anticancer agent, it is useful in the treatment of diseases associated with abnormal cell proliferation such as cancer.

  In the general formula (1), R is a phenyl group which may be unsubstituted or substituted, a benzyl group which may be unsubstituted or substituted, or a substituent which is unsubstituted or substituted. Any of the phenethyl groups may be used, but for example, 4-methylbenzyl group, 4-tert-butylbenzyl group, 4-methoxybenzyl group, 4-phenylbenzyl group, 4-phenoxybenzyl group and the like are preferable.

In the general formula (1), X represents O, CH ₂ or S, Y represents CH ₂ , XY may be a double bond or a triple bond, and Z represents N or CH. Is preferably O, Y is preferably CH ₂ and Z is preferably N.

  In the general formula (1), R ′ represents a hydrogen atom or a lower alkyl group, and for example, a hydrogen atom is preferable. Here, the “lower alkyl group” may be linear, branched, cyclic, or a combination thereof having 1 to 14 carbon atoms, preferably 1 to 10 carbon atoms.

The salts of the compound represented by the general formula (1) in the present invention are conventional, and include metal salts such as alkali metal salts (for example, sodium salts, potassium salts and lithium salts), alkaline earth metal salts (for example, calcium). And pharmaceutically acceptable salts such as aluminum salts and the like.
Moreover, unless otherwise indicated, the compound represented by General formula (1) in this invention also includes the geometric isomers (for example, E body, Z body, etc.) and optical isomers.

Although it does not limit as a compound and its salt shown by General formula (1), For example, the following are mentioned.
6- (3-hydroxy-2-oxopropoxy) -2- (4′-methylbenzyl) isoindoline-1-one and salts thereof,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-tert-butylbenzyl) isoindoline-1-one and salts thereof,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-methoxybenzyl) isoindoline-1-one and salts thereof,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-phenylbenzyl) isoindoline-1-one and salts thereof,
6- (3-hydroxy-2-oxopropoxy) -2- (4'-phenoxybenzyl) isoindoline-1-one and its salts.

は例えば以下の方法により製造することができる(スキーム 1)。 Among the compounds represented by the general formula (1) of the present invention, in the formula, R is an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group. A phenethyl group which may be unsubstituted or substituted, X is O, Y is CH ₂ , Z is N, and R ′ is a hydrogen atom (1a)

Can be produced, for example, by the following method (Scheme 1).

［式中式中、Ｒは無置換又は置換基を有していても良いフェニル基、無置換または置換基を有していても良いベンジル基、無置換または置換基を有していても良いフェネチル基である］で示される化合物は公知化合物である５−ヒドロキシ−２−メチル安息香酸メチル（化合物２）の水酸基をｔ−ブチルジメチルシリル基（ＴＢＳ）で保護（第一工程）することにより合成される５−ｔ−ブチルジメチルシリルオキシ−２−メチル安息香酸メチル（化合物３）のメチル基を臭素化して（第二工程）得られる２−ブロモメチル−５−ｔ−ブチルジメチルシリルオキシ安息香酸メチル（化合物４）に一般式（１１）

［式中、Ｒは無置換又は置換基を有していても良いフェニル基、無置換または置換基を有していても良いベンジル基、無置換または置換基を有していても良いフェネチル基を表す］で表される化合物を反応させる（第三工程）ことにより得られる一般式（５）

［式中、Ｒは前述の通りである］で表される化合物を脱保護（第四工程）することにより得られる一般式（６）

［式中、Ｒは前述の通りである］で表される化合物にアリルブロマイドを反応させる（第五工程）ことにより得られる一般式（７）

［式中、Ｒは前述の通りである］で表される化合物をオスミウム酸化（第六工程）することにより得られる一般式（８）

［式中、Ｒは前述の通りである］で表される化合物の一級水酸基をＴＢＳで保護（第七工程）することにより得られる一般式（９）

［式中、Ｒは前述の通りである］で表される化合物の二級水酸基を酸化（第八工程）することによりことにより得られる一般式（１０）

［式中、Ｒは前述の通りである］で表される化合物のＴＢＳ基を脱保護（第九工程）することにより製造することができる。 That is, the general formula (1a)

[In the formula, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. Is a group obtained by protecting the hydroxyl group of methyl 5-hydroxy-2-methylbenzoate (Compound 2), which is a known compound, with a t-butyldimethylsilyl group (TBS) (first step). 2-bromomethyl-5-t-butyldimethylsilyloxybenzoate obtained by brominating the methyl group of methyl 5-t-butyldimethylsilyloxy-2-methylbenzoate (compound 3) (second step) (Compound 4) is represented by the general formula (11)

[Wherein, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. General formula (5) obtained by reacting the compound represented by

[Wherein R is as defined above] General formula (6) obtained by deprotection (fourth step)

[In the formula, R is as defined above] General formula (7) obtained by reacting allyl bromide with the compound represented by (5th step)

[Wherein R is as defined above] General formula (8) obtained by osmium oxidation (sixth step)

[Wherein R is as defined above] The primary hydroxyl group of the compound represented by the general formula (9) obtained by protecting with TBS (seventh step)

[Wherein, R is as defined above] General formula (10) obtained by oxidizing (eighth step) the secondary hydroxyl group of the compound represented by

[Wherein, R is as defined above] can be produced by deprotecting the TBS group (9th step).

The reaction in the first step can be carried out in a solvent such as N, N-dimethylformamide or dimethyl sulfoxide. The reaction can be carried out at -50 ° C to 200 ° C, preferably 0 ° C to 100 ° C.
The reaction in the second step can be carried out by heating using a brominating agent in a carbon tetrachloride solvent and under irradiation or non-irradiation of light from a tungsten lamp or a halogen lamp. The reaction can be carried out at 0 to 150 ° C., preferably 50 to 100 ° C. As the brominating agent, bromine, N-bromosuccinimide and the like can be used.
The reaction in the third step is performed in a solvent such as methanol, ethanol, toluene, methylene chloride, tetrahydrofuran, dioxane, N, N-dimethylformamide as a base, for example, an alkali metal hydride such as sodium hydride, or sodium hydroxide. It can be carried out in the presence or absence of an alkali metal carbonate, an alkali metal carbonate such as potassium carbonate, or an organic base such as pyridine or triethylamine. The reaction can be performed at -20 ° C to 150 ° C, preferably 0 ° C to 100 ° C.

The reaction in the fourth step can be carried out in a solvent such as tetrahydrofuran, toluene, dioxane, N, N-dimethylformamide and the like in the presence of tertbutylammonium fluoride. The reaction can be performed at -20 ° C to 150 ° C, preferably 0 ° C to 100 ° C.
The reaction in the fifth step is carried out in a solvent such as tetrahydrofuran, toluene, dioxane, N, N-dimethylformamide as a base, for example, an alkali metal hydride such as sodium hydride, an alkali metal hydroxide such as sodium hydroxide, carbonic acid. It can be carried out in the presence of an alkali metal carbonate such as cesium or potassium carbonate. The reaction temperature can be 0 to 200 ° C., preferably 0 to 100 ° C.
The reaction in the sixth step can be carried out in the presence of AD-mix-α in a mixed solvent of water and tert-butanol. The reaction can be carried out at -50 ° C to 200 ° C, preferably 0 ° C to 100 ° C.
The reaction in the seventh step can be performed in a solvent such as methylene chloride or chloroform in the presence of 4-dimethylaminopyridine, 4-pyrrolidinopyridine, diisopropylethylamine, or triethylamine as a base. The reaction can be carried out at -50 ° C to 200 ° C, preferably 0 ° C to 100 ° C.
The reaction in the eighth step can be carried out in a solvent such as methylene chloride and chloroform in the presence of desmartin periodate, pyridinium dichromate, and pyridinium chlorochromate as oxidizing agents. The reaction can be carried out at -50 ° C to 200 ° C, preferably 0 ° C to 100 ° C.
The reaction in the ninth step can be carried out using acetic acid, trifluoroacetic acid, trifluoromethanesulfonic acid, camphorsulfonic acid, etc. as an acid in tetrahydrofuran, ethanol, acetone, dioxane and mixed solvents of these organic solvents and water. . The reaction temperature can be 0 to 200 ° C., preferably room temperature to 100 ° C.

Since the compound represented by the general formula (1) of the present invention selectively inhibits HDAC6 activity, it can be used as a therapeutic agent for diseases caused by the enhancement of HDAC6 activity. Examples of diseases caused by increased HDAC6 activity include cancer and neurodegenerative diseases. Here, the cancer that can be mentioned as a treatment target of the compound represented by the general formula (1) of the present invention is not limited, but includes brain tumor, neuroblastoma, lung cancer, stomach cancer, colon cancer, rectal cancer, liver cancer, Pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, leukemia (especially acute promyelocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia, acute lymphoblastic leukemia, chronic Lymphoid leukemia), myeloma, osteosarcoma, myeloma and the like.
Further, the neurodegenerative diseases that can be treated with the compound represented by the general formula (1) of the present invention are not limited, but include bulbar spinal muscular atrophy, Huntington's disease, spinocerebellar degeneration type 1 (SCA1). ), Spinocerebellar degeneration type 2 (SCA2), Machado-Joseph disease (MJD or SCA3), spinocerebellar degeneration type 6 (SCA6), spinocerebellar degeneration type 7 (SCA7) or dentate red nucleus / pale Examples include Parkinson's disease, Alzheimer's disease, etc. in addition to polyglutamine diseases such as Ryukyu body atrophy (DRPLA).

The compound of the present invention can be used as a medicament in the form of a pharmaceutical composition that does not adversely affect the living body. Such compositions typically include a pharmacologically acceptable carrier in addition to the compound of the present invention.
“Pharmaceutically acceptable carrier” refers to those suitable for pharmaceutical administration, including solvents, dispersion media, coating agents, antibacterial and antifungal agents, agents that act isotonically to delay adsorption and the like. . Examples of such carriers and those preferred for diluting the carriers include, but are not limited to, water, saline, finger solutions, dextrose solutions, and human serum albumin. Non-aqueous media such as liposomes and non-volatile oils are also used. In addition, certain compounds that protect or promote the activity of the compounds of the present invention may be included in the composition.

The medicament of the present invention includes intravenous, intradermal, subcutaneous, oral (including inhalation, etc.), transdermal and transmucosal administration, and is formulated to be suitable for a therapeutically appropriate route of administration. . Solutions or suspensions used for parenteral, intradermal, or subcutaneous application include, but are not limited to, sterile diluents such as water for injection, saline solutions, non-volatile oils, polyethylene glycols, Glycerin, propylene glycol, or other synthetic solvents, benzyl alcohol or other preservatives such as methylparaben, antioxidants such as ascorbic acid or sodium bisulfite, soothing agents such as benzalkonium chloride, procaine hydrochloride, ethylenediaminetetraacetic acid Chelating agents such as (EDTA), buffering agents such as acetate, citrate, or phosphate, and agents for osmotic pressure adjustment such as sodium chloride or dextrose.
The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. Parenteral preparations are contained in ampoules, glass or plastic disposable syringes or multiple dose vials.

  Pharmaceutical compositions suitable for injection include sterile aqueous solutions (water soluble) or dispersion media and sterile powders for the preparation of sterile injectable solutions or dispersion media at the time of use. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, or phosphate buffered saline (PBS). When used as an injection, the composition must be sterile and must be fluid enough to be administered with a syringe. The composition must be stable to chemical changes, corrosion, and the like during formulation and storage, and must prevent contamination from microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures. For example, using a coating agent such as lectin, maintaining a required particle size in the dispersion medium, and maintaining a proper fluidity by using a surfactant. Various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, and thimerosal can be used to prevent microbial contamination. In addition, polyalcohols such as sugar, mannitol, sorbitol, and agents that maintain isotonicity such as sodium chloride may be included in the composition. Compositions that can delay adsorption include agents such as aluminum monostearate and gelatin.

  Sterile injectable solutions are prepared by adding the required ingredients alone or in combination with other ingredients to the required amount of the active compound in a suitable solvent and sterilizing. Generally, a dispersion medium is prepared by incorporating the active compound into a sterile medium that contains a basic dispersion medium and the other necessary ingredients described above. Methods for preparing a sterile powder for preparing a sterile injectable solution include vacuum drying and lyophilization to prepare a powder containing the active ingredient and any desired ingredients derived from the sterile solution. .

Oral compositions include inert diluents or carriers that are not harmful when incorporated into the body. Oral compositions are, for example, contained in gelatin capsules or compressed into tablets. For oral treatment, the active compound is incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a flowable carrier, and the composition in the flowable carrier is applied orally. In addition, pharmaceutically compatible binding agents, and / or adjuvant materials may be included.
Tablets, pills, capsules, troches and the like may contain any of the following components or compounds with similar properties: excipients such as microcrystalline cellulose, binding such as gum arabic, tragacanth or gelatin Agents; swelling agents such as alginic acid, PRIMOGEL, or corn starch such as starch or lactose; lubricants such as magnesium stearate or STRROTES; lubricants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or peppermint, methyl A fragrance additive such as salicylic acid or orange flavor.

  The compounds of the present invention can be prepared as sustained release formulations such as delivery systems encapsulated in implantable tablets and microcapsules using a carrier that can prevent immediate removal from the body. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Such materials can be readily prepared by those skilled in the art. Liposome suspensions can also be used as pharmaceutically acceptable carriers. Useful liposomes are prepared as a lipid composition comprising, but not limited to, phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanol (PEG-PE) through a filter of appropriate pore size so as to be of a size suitable for use. Purified by evaporation.

In the treatment or prevention of a specific disease with the compound of the present invention, the appropriate dosage level depends on the condition of the patient to be administered, the administration method, etc., but can be easily optimized by those skilled in the art. It is.
In the case of injection administration, for example, it is preferable to administer about 0.1 μg / kg to about 500 mg / kg of the patient's body weight per day, and it will generally be possible to administer a single dose or divided into multiple doses. Preferably, the dosage level is about 0.1 μg / kg to about 250 mg / kg per day, more preferably about 0.5 to about 100 mg / kg per day.
For oral administration, the composition is preferably provided in the form of a tablet containing 1.0 to 1000 mg of active ingredient, preferably 1.0, 5.0, 10.0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750. 0, 800.0, 900.0 and 1000.0 mg. The compounds are administered on a regimen of 1 to 4 times daily, preferably once or twice daily.

  A pharmaceutical composition or formulation must consist of uniform unit doses to ensure a constant dose. A unit dose is a unit formulated with a pharmaceutically acceptable carrier, including a single dose effective for treating a patient. When determining the unit dosage of the present invention, physical and chemical characteristics of the compound to be formulated, expected therapeutic effects, considerations specific to the compound, and the like are considered.

    Furthermore, when the compound of the present invention is administered in combination with a drug exhibiting anticancer activity, a remarkable anticancer effect or anticancer effect is expected. Therefore, when the medicament of the present invention is used for the purpose of anticancer or anticancer, one or more of other anticancer or anticancer agents may be further added. Here, the anticancer agent or anticancer agent to be used is not limited, but for example, differentiation induction therapy such as taxane-based docetaxel, paclitaxel or leukemia (especially acute promyelocytic leukemia) acting on microtubules. All-trans retinoic acid (ATRA) used can be used.

  The pharmaceutical composition of the present invention can be included in the form of a kit in a container or pack together with instructions for administration. When the pharmaceutical composition according to the present invention is supplied as a kit, different constituents of the pharmaceutical composition are packaged in separate containers and mixed immediately before use. The reason why the components are packaged separately in this way is to enable long-term storage without losing the function of the active component.

  Reagents contained in the kit are supplied in any type of container in which the components remain active for an extended period of time, are not adsorbed by the container material, and are not subject to alteration. For example, a sealed glass ampoule includes a buffer packaged under a neutral and non-reactive gas such as nitrogen gas. Ampoules are composed of glass, polycarbonate, organic polymers such as polystyrene, ceramics, metals, or any other suitable material commonly used to hold reagents. Examples of other suitable containers include simple bottles made from similar materials such as ampoules, and packaging materials that are internally lined with foil such as aluminum or an alloy. Other containers include test tubes, vials, flasks, bottles, syringes, or the like. The container has a sterile access port such as a bottle having a stopper that can be penetrated by a hypodermic needle.

  The kit also includes instructions for use. Instructions for using the kit comprising the pharmaceutical composition are printed on paper or other material and / or floppy disk, CD-ROM, DVD-ROM, Zip disk, video tape, audio tape, etc. It may be supplied as an electrically or electromagnetically readable medium. Detailed instructions for use may be actually attached to the kit, or may be posted on a website designated by the manufacturer or distributor of the kit or notified by e-mail or the like.

  Furthermore, the present invention includes a method for treating a mammal suffering from a disease that develops due to enhanced HDAC6 activity. Here, “treatment” means to prevent or alleviate the progression and worsening of the disease state in a mammal that is likely to suffer from or suffers from the disease. Used to mean therapeutic treatment aimed at preventing or alleviating progression and worsening.

The term “disease” means any disease that develops due to increased HDAC6 activity, and is not limited to, for example, brain tumor, neuroblastoma, lung cancer, stomach cancer, colon cancer, rectal cancer, Liver cancer, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, leukemia (especially acute promyelocytic leukemia, acute myeloid leukemia, chronic myelogenous leukemia, acute lymphocytic leukemia) , Chronic lymphocytic leukemia, etc.), myeloma, osteosarcoma, myeloma and other cancers, or bulbar spinal muscular atrophy, Huntington's disease, spinocerebellar degeneration type 1 (SCA1), spinocerebellar degeneration type 2 (SCA2), Polyglucose such as Machado-Joseph disease (MJD or SCA3), spinocerebellar degeneration type 6 (SCA6), spinocerebellar degeneration type 7 (SCA7), or dentate nucleus red nucleus and pallidal louis atrophy (DRPLA) Other Min Disease, Parkinson's disease, such as Alzheimer's disease and the like neurodegenerative diseases.
Particularly in the treatment of cancer, the compound of the present invention shows a remarkable effect when used in combination with other anticancer agents or anticancer agents. Therefore, the compound of the present invention is used in combination with other anticancer agents or anticancer agents. It is desirable. Here, other anticancer agents or anticancer agents are not limited. For example, they are used for differentiation induction therapy such as taxane-based docetaxel, paclitaxel, or leukemia (especially acute promyelocytic leukemia) acting on microtubules. All-trans retinoic acid (ATRA) that can be used can be used.

  Next, the present invention will be described with reference to specific examples, but the present invention is not limited to these examples.

６−（２−ヒドロキシ−３−ｔｅｒｔ−ブチルジメチルシリロキシ）−２−（４’−メチルベンジル）−イソインドリン−１−オン（５８ｍｇ，０．１３２ｍｍｏｌ）のＣＨ_２Ｃｌ_２（２ｍＬ）溶液にＤＭＰ（１４０ｍｇ，０．３３０ｍｍｏｌ）を加え、室温で一晩撹拌した。ＡｃＯＥｔで抽出、Ｈ_２Ｏ・飽和食塩水で洗浄、乾燥（ＭｇＳＯ_４）した。溶媒を留去し、ＣＨ_３ＣＯＯＨ／Ｈ_２Ｏ／ＴＨＦ（３ｍＬ／１ｍＬ／１ｍＬ）に溶かし、一晩撹拌した。ＡｃＯＥｔで抽出、Ｈ_２Ｏ・飽和食塩水で洗浄、乾燥（ＭｇＳＯ_４）した。溶媒を留去し、残留物をシリカゲルクロマトグラフィー（Ｈ：Ａ ＝１：２→Ａ）で精製し、標題化合物を白色固体として得た。
収量：４２ｍｇ（９８％）。
NMR(CDCl₃) ppm:δ7.33(d, J = 2.6 Hz, 1H), 7.31(d, J = 8.1 Hz, 1H), 7.19(d, J = 7.7 Hz, 2H), 7.14(d, J = 7.7 Hz, 2H), 7.13(dd, J = 2.6, 8.1 Hz, 1H), 4.79(s, 2H), 4.76(s, 2H), 4.59(s, 2H), 4.21(s, 2H), 2.33(s, 3H).
FAB-MS:C₁₉H₁₉NO₄, exact mass:325.13, found:326(M+H)⁺. Example 1 6- (3-Hydroxy-2-oxopropoxy) -2- (4′-methylbenzyl) isoindoline-1-one

To a solution of 6- (2-hydroxy-3-tert-butyldimethylsilyloxy) -2- (4′-methylbenzyl) -isoindoline-1-one (58 mg, 0.132 mmol) in CH ₂ Cl ₂ (2 mL). DMP (140 mg, 0.330 mmol) was added and stirred overnight at room temperature. Extracted with AcOEt, washed with H ₂ O.saturated brine, and dried (MgSO ₄ ). The solvent was distilled off and dissolved in CH ₃ COOH / H ₂ O / THF (3 mL / 1 mL / 1 mL) and stirred overnight. Extracted with AcOEt, washed with H ₂ O.saturated brine, and dried (MgSO ₄ ). The solvent was distilled off, and the residue was purified by silica gel chromatography (H: A = 1: 2 → A) to give the title compound as a white solid.
Yield: 42 mg (98%).
NMR (CDCl ₃ ) ppm: δ 7.33 (d, J = 2.6 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 7.19 (d, J = 7.7 Hz, 2H), 7.14 (d, J = 7.7 Hz, 2H), 7.13 (dd, J = 2.6, 8.1 Hz, 1H), 4.79 (s, 2H), 4.76 (s, 2H), 4.59 (s, 2H), 4.21 (s, 2H), 2.33 (s, 3H).
FAB-MS: C ₁₉ H ₁₉ NO ₄ , exact mass: 325.13, found: 326 (M + H) ⁺ .

６−（２−ヒドロキシ−３−ｔｅｒｔ−ブチルジメチルシリロキシ）−２−（４’−ｔｅｒｔブチルベンジル）イソインドリン−１−オン（７５ｍｇ，０．１５５ｍｍｏｌ）を原料に、実施例１と同様の方法で標題化合物を白色固体として得た。
収量：４８ｍｇ（８４％）．
NMR（CDCl₃) ppm：δ7.35-7.31（m, 4H), 7.23（d, J = 8.5 Hz, 2H), 7.13（dd, J = 2.6, 8.5 Hz, 1H), 4.80（s, 2H), 4.76（s, 2H), 4.59（s, 2H), 4.23（s, 2H), 1.30（s, 9H).
FAB-MS：C₂₂H₂₅NO₄, exact mass：367.18, found：368（M+H)⁺. Example 2 6- (3-Hydroxy-2-oxopropoxy) -2- (4′-tertbutylbenzyl) isoindoline-1-one

6- (2-Hydroxy-3-tert-butyldimethylsilyloxy) -2- (4′-tertbutylbenzyl) isoindolin-1-one (75 mg, 0.155 mmol) was used as a starting material in the same manner as in Example 1. The method gave the title compound as a white solid.
Yield: 48 mg (84%).
NMR (CDCl ₃ ) ppm: δ 7.35-7.31 (m, 4H), 7.23 (d, J = 8.5 Hz, 2H), 7.13 (dd, J = 2.6, 8.5 Hz, 1H), 4.80 (s, 2H) , 4.76 (s, 2H), 4.59 (s, 2H), 4.23 (s, 2H), 1.30 (s, 9H).
FAB-MS: C ₂₂ H ₂₅ NO ₄ , exact mass: 367.18, found: 368 (M + H) ⁺ .

６−（２−ヒドロキシ−３−ｔｅｒｔ−ブチルジメチルシリロキシ）−２−（４’−メトキシベンジル）イソインドリン−１−オン（１２０ｍｇ，０．２６３ ｍｍｏｌ）を原料に、実施例１と同様の方法で標題化合物を白色固体として得た。
収量：７３ ｍｇ（８１％）．
NMR（CDCl₃) ppm：δ7.32（d, J = 2.6 Hz, 1H), 7.31（d, J = 8.6 Hz, 1H), 7.22（d, J = 8.5 Hz, 2H), 7.13（dd, J = 2.6, 8.6 Hz, 1H), 6.86（d, J = 8.5 Hz, 2H), 4.80（s, 2H), 4.73（s, 2H), 4.58（s, 2H), 4.20（s, 2H), 3.79（s, 3H).
FAB-MS：C₁₉H₁₉NO₅, exact mass：341.13, found：342（M+H)⁺. Example 3 6- (3-Hydroxy-2-oxopropoxy) -2- (4′-methoxybenzyl) isoindoline-1-one

Using 6- (2-hydroxy-3-tert-butyldimethylsilyloxy) -2- (4′-methoxybenzyl) isoindoline-1-one (120 mg, 0.263 mmol) as a starting material, the same as in Example 1 The method gave the title compound as a white solid.
Yield: 73 mg (81%).
NMR (CDCl ₃ ) ppm: δ 7.32 (d, J = 2.6 Hz, 1H), 7.31 (d, J = 8.6 Hz, 1H), 7.22 (d, J = 8.5 Hz, 2H), 7.13 (dd, J = 2.6, 8.6 Hz, 1H), 6.86 (d, J = 8.5 Hz, 2H), 4.80 (s, 2H), 4.73 (s, 2H), 4.58 (s, 2H), 4.20 (s, 2H), 3.79 (S, 3H).
FAB-MS: C ₁₉ H ₁₉ NO ₅ , exact mass: 341.13, found: 342 (M + H) ⁺ .

６−（２−ヒドロキシ−３−ｔｅｒｔ−ブチルジメチルシリロキシ）−２−（４’−フェニルベンジル）イソインドリン−１−オン（２８ｍｇ，０．０５４０ｍｍｏｌ）を原料に、実施例１と同様の方法で標題化合物を無色油状物として得た。
収量：１５ｍｇ（７２％）．
NMR（CDCl₃) ppm：δ7.56（d, J = 7.7 Hz, 2H), 7.56 （d, J = 7.7 Hz, 2H), 7.43（t, J = 7.7 Hz, 2H), 7.37-7.32（m, 5H), 7.15（dd, J = 2.1, 8.1 Hz, 1H), 4.84（s, 2H), 4.81（s, 2H), 4.59（s, 2H), 4.27（s, 2H).
FAB-MS：C₂₄H₂₁NO₄, exact mass：387.15, found：388（M+H)⁺. Example 4 6- (3-Hydroxy-2-oxopropoxy) -2- (4′-phenylbenzyl) isoindoline-1-one

6- (2-Hydroxy-3-tert-butyldimethylsilyloxy) -2- (4′-phenylbenzyl) isoindoline-1-one (28 mg, 0.0540 mmol) as a starting material, the same method as in Example 1 Gave the title compound as a colorless oil.
Yield: 15 mg (72%).
NMR (CDCl ₃ ) ppm: δ 7.56 (d, J = 7.7 Hz, 2H), 7.56 (d, J = 7.7 Hz, 2H), 7.43 (t, J = 7.7 Hz, 2H), 7.37-7.32 (m , 5H), 7.15 (dd, J = 2.1, 8.1 Hz, 1H), 4.84 (s, 2H), 4.81 (s, 2H), 4.59 (s, 2H), 4.27 (s, 2H).
FAB-MS: C ₂₄ H ₂₁ NO ₄ , exact mass: 387.15, found: 388 (M + H) ⁺ .

６−（２−ヒドロキシ−３−ｔｅｒｔブチルジメチルシリロキシ）−２−（４’−フェノキシベンジル）イソインドリン−１−オン（１４７ｍｇ，０．２８３ｍｍｏｌ）を原料に、実施例１と同様の方法で標題化合物を白色固体として得た。
収量：８４ ｍｇ（７４％）．
NMR(CDCl₃) ppm:δ7.34-7.31(m, 3H), 7.27?7.25(m, 3H), 7.14(dd, J = 2.6, 8.1 Hz, 1H), 7.10(t, J = 7.5 Hz, 1H), 6.99(d, J = 7.5 Hz, 2H), 6.97(d, J = 7.5 Hz, 2H), 4.79(s, 2H), 4.77 (s, 2H), 4.59(s, 2H), 4.24(s, 2H).
FAB-MS:C24H21NO5, exact mass:403.14, found:404(M+H)⁺. Example 5 6- (3-Hydroxy-2-oxopropoxy) -2- (4′-phenoxybenzyl) isoindoline-1-one

In the same manner as in Example 1, using 6- (2-hydroxy-3-tertbutyldimethylsilyloxy) -2- (4′-phenoxybenzyl) isoindoline-1-one (147 mg, 0.283 mmol) as a raw material. The title compound was obtained as a white solid.
Yield: 84 mg (74%).
_{NMR (CDCl 3) ppm:?} Δ7.34-7.31 (m, 3H), 7.27 7.25 (m, 3H), 7.14 (dd, J = 2.6, 8.1 Hz, 1H), 7.10 (t, J = 7.5 Hz, 1H), 6.99 (d, J = 7.5 Hz, 2H), 6.97 (d, J = 7.5 Hz, 2H), 4.79 (s, 2H), 4.77 (s, 2H), 4.59 (s, 2H), 4.24 ( s, 2H).
FAB-MS: C24H21NO5, exact mass: 403.14, found: 404 (M + H) ⁺ .

[Experimental Example 1] HDAC Inhibitory Activity A HDAC reaction solution used for measurement of HDAC inhibitory activity was prepared as follows.
A 293T cell (1 × 10 ⁷ cells) was seeded in a 100 mm dish, and 24 hours later, a vector (1 μg) expressing human HDAC1, 4 or mouse HDAC6 was transfected using LipofectAmine 2000 reagent (Life Technologies, Inc.). Here, the above HDAC1 expression vector is pcDNA3-HD1 (Yang et al., J. Biol. Chem., 272: 2801-28007, 1997), and the human HDAC4 expression vector is pcDNA3.1 (+)-HD4 (Fischle et al., J. Biol. Chem., 274: 11713-11720, 1999), pcDNA-mHDA2 / HDAC6 (Verdel et al., J. Biol. Chem., 274: 2440-2445, 1999) was used as the mouse HDAC6 expression vector.
The cells were collected by incubation in Dulbecco's modified Eagles's medium (DMEM) for 24 hours, washed with PBS, and then lys buffer (50 mM Tris-HCl (pH 7.5), 120 mM NaCl, 5 mM EDTA, 0. 5% Nonidet P-40) and sonicated. Supernatants were collected by centrifugation and non-specific proteins were removed using protein A / G plus agarose beads (Santa Cruz Biotechnologies, Inc.). Then, anti-FLAG M2 antibody (Sigma-Aldrich Inc.) was added and reacted at 4 ° C. for 2 hours. Agarose beads (protein A / G plus agarose beads (Santa Cruz Biotechnologies, Inc.)) were added to this reaction solution, and the mixture was further reacted at 4 ° C. for 3 hours, and then the agarose beads were washed three times with a lysis buffer. Washed once with buffer (20 mM Tris-HCl (pH 8.0), 150 mM NaCl, 10% glycerol). Incubate with FLAG peptide (40 μg) (Sigma-Aldrich Inc.) in HD buffer (200 μl) at 4 ° C. for 1 hour to recover the protein bound from the agarose beads, and use it as an HDAC reaction solution for the following HDAC inhibitory activity measurement. It was.

The inhibitory activity of HDAC1, HDAC4 and HDAC6 was evaluated as follows. The representative compounds shown in Examples 1 to 5 were dissolved in DMSO to prepare a 10 mM concentration stock solution, which was used as the inhibitor stock solution. The assay was performed by incubating an HDAC solution and an acetylated histone peptide solution labeled with coumarin at 37 ° C. for 30 minutes in the presence of a test compound (reaction solution 20 μl). 30 μl trypsin was added to the reaction solution, and aminomethylcoumarin cleaved by the enzyme reaction was measured with a fluorescent plate reader. As a negative control, the same operation was performed without adding an inhibitor to the reaction system. The inhibitory activity was expressed as a 50% inhibitory concentration (“IC ₅₀ (μM)”) of HDAC activity in the negative control (Table 1). From this result, it was revealed that the compound of the present invention exhibits selective inhibitory activity against HDAC6.

[Experimental Example 2] Differentiation Induction Promoting Activity All-trans retinoic acid (ATRA) is an active vitamin A metabolite that induces cell differentiation and is actually clinically applied as a therapeutic agent for leukemia. On the other hand, HDAC inhibitors are known to have cell differentiation-inducing activity, and when used in combination with ATRA, the amount of ATRA used can be reduced and side effects are expected to be reduced. Therefore, human promyelocytic leukemia cells (HL-60) were used and the differentiation induction promoting activity of the compound shown in Example 1 of the present invention was evaluated in the presence of ATRA 0.6 nM. As a result, it was clarified that the compound shown in Example 1 of the present invention exhibits cell differentiation-inducing activity, similar to Zolinza ^™ (Merck) known as an HDAC inhibitor (FIG. 1). ).

  Since the compound of the present invention selectively inhibits HDAC6 activity, it can be used for the treatment of HDAC6-related diseases and the development of therapeutic agents therefor.

FIG. 5 shows the cell differentiation induction promoting activity in the presence of ATRA by the compound shown in Example 1 of the present invention. FIG. The control indicates the differentiation induction rate of cells when only DMSO used as a solvent for the compound is added in the presence of 0.6 nM ATRA and DMSO in the presence of 0.6 nM ATRA.

Claims (10)

General formula (1)

[Wherein, R represents an unsubstituted or substituted phenyl group, an unsubstituted or substituted benzyl group, an unsubstituted or substituted phenethyl group. Represents a group, X represents O, CH ₂ or S, Y represents CH ₂ , XY may be a double bond or a triple bond, Z represents N or CH, R ′ represents a hydrogen atom, lower A hydroxymethyl ketone derivative represented by the formula: or a pharmacologically acceptable salt thereof, or a hydrate thereof. X is O, Y is CH _{2, Z} is hydroxymethyl ketone derivative or a pharmacologically acceptable salt thereof or a hydrate thereof according to claim 1 N and R 'is a hydrogen atom. The hydroxymethyl ketone derivative or a pharmacologically acceptable salt thereof or a hydrate thereof according to claim 2, wherein R is a substituent represented by any of the following formulas (101) to (105).

The hydroxymethyl ketone derivative is
6- (3-hydroxy-2-oxopropoxy) -2- (4′-methylbenzyl) isoindoline-1-one,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-tert-butylbenzyl) isoindoline-1-one,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-methoxybenzyl) isoindoline-1-one,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-phenylbenzyl) isoindoline-1-one,
6- (3-hydroxy-2-oxopropoxy) -2- (4′-phenoxybenzyl) isoindoline-1-one,
The hydroxymethyl ketone derivative according to claim 1 or a pharmacologically acceptable salt thereof or a hydrate thereof selected from the group consisting of:   An HDAC6 inhibitor comprising the hydroxymethyl ketone derivative according to any one of claims 1 to 4 or a pharmacologically acceptable salt thereof or a hydrate thereof, and a pharmacologically acceptable carrier.   5. Enhancement of HDAC6 activity comprising the hydroxymethyl ketone derivative or pharmacologically acceptable salt thereof or hydrate thereof according to claim 1, and a pharmacologically acceptable carrier. A medicament for the treatment of the disease caused.   The medicament according to claim 6, wherein the disease is cancer.   The medicament according to claim 7, further comprising an effective amount of an anticancer agent or an anticancer agent.   The medicament according to claim 6, wherein the disease is a neurodegenerative disease.   The medicament according to claim 9, wherein the disease is polyglutamine disease.

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