KR20240012846A - Novel HDAC6-selective inhibitors and uses thereof - Google Patents

Abstract

The present invention relates to a novel HDAC6 selective inhibitor and its use. A compound capable of selectively inhibiting HDAC6 has been optimized to present a new HDAC6 inhibitor base structure, which not only has excellent HDAC6 inhibitory activity effect, but also has other downstream effects. Since it can act selectively and effectively on HDAC6 compared to HDAC, it can be useful as a composition for preventing or treating HDAC6-mediated diseases.

Description

Novel HDAC6-selective inhibitors and uses thereof}

The present invention relates to novel HDAC6 selective inhibitors and uses thereof.

Histone deacetylase (HDAC) is an enzyme that removes acetyl groups from specific histone protein residues in promoter and enhancer regions, an essential part of the regulation of cellular gene transcription, causing cell-cycle arrest in the G1 and/or G2 phases. It performs the function of controlling gene expression. In addition, it also regulates gene expression indirectly by mediating acetylation of non-histone proteins such as DNA-binding proteins, transcription factors, signaling factors, DNA repair and chaperone proteins.

HDAC inhibitors exhibit anticancer efficacy by selectively inducing the expression of epigenetically inhibited tumor suppressor genes through transcription-dependent/independent mechanisms involved in acetylation of histone and non-histone proteins. It plays a role. It can cause growth arrest with subsequent differentiation or apoptosis of tumor cells, without any effect on normal cells.

The growth inhibitory effect of HDAC inhibitors has been confirmed in vitro in virtually all transformed cell types, including cell lines originating from both hematological and epithelial tumors, breast, prostate, lung and gastric cancer, and neuroblastoma. Vorinostat (suberoylanilide hydroxamic acid (SAHA)), which was the first to receive clinical approval in addition to being proven in animal models of various cancer types such as leukemia, showed positive effects in patients with some solid or hematologic malignancies.

However, there are limitations such as the anti-cancer efficacy of HDAC inhibitors not being significantly superior to that of other anti-cancer drugs, side effects being observed, resistance to drugs, and low utilization of HDAC inhibitors other than anti-cancer effects. In addition, most HDAC inhibitors approved for clinical use are pan-HDAC inhibitors with no selectivity between subtypes, and the utilization and application of subtype-specific HDAC inhibitors is lacking.

Meanwhile, 12 subtypes of HDAC have been reported, and each subtype has different functions and roles, and is known to exhibit differences in associated diseases. While most HDACs exist inside the nucleus and are involved in histone and intranuclear deacetylation, HDAC6 moves between the nucleus and the cytoplasm and is mainly involved in the deacetylation of proteins present in the cytoplasm. Inhibition of HDAC6 activity has been reported to have anti-cancer effects, immune response suppression effects, and therapeutic effects on neurodegenerative diseases. Therefore, HDAC6 selective inhibitors are used in the development of new mechanisms for treating immune diseases and neurodegenerative diseases, as well as in the application and development of subtype-selective HDAC inhibitors. You can contribute.

To date, HDAC6-selective inhibitors are mainly undergoing clinical trials as anticancer drugs, and as it has recently been reported that inhibiting HDAC6 has therapeutic effects on immune response suppression and neurodegenerative diseases, the need to develop effective HDAC6-selective inhibitors has increased. Research is being actively conducted to develop HDAC6 selective inhibitors for indications.

Republic of Korea Patent Publication No. 10-2016-0043534

In order to solve the above problems, the present invention sought to develop an HDAC6 inhibitor that can selectively and effectively inhibit HDAC6 among HDAC subtypes.

Accordingly, one object of the present invention is to provide a compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In the above formula, R is,

, , , , , , , , , , and is any one selected from the group consisting of, wherein X ₁ is any one selected from the group consisting of H, F, Br and OMe, where X ₂ is H or F, and where n is 0, 2, or 3 It is an integer.

In addition, another object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of HDAC6-mediated diseases comprising the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In addition, another object of the present invention is to provide a method for preventing or treating HDAC6-mediated diseases, including the step of administering the composition.

However, the technical problem to be achieved by the present invention is not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below. There will be.

In order to achieve the above object, the present invention provides a compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In the above formula, R is,

, , , , , , , , , , and is any one selected from the group consisting of, wherein X ₁ is any one selected from the group consisting of H, F, Br and OMe, where X ₂ is H or F, and where n is 0, 2, or 3 It is an integer.

In another embodiment of the present invention, the compound is N-hydroxy-3-(2-(naphthalen-1-yl)thiazol-4-yl)propanamide (20), N-hydroxy-2- (naphthalen-2-yl)thiazol-4-carboxamide (21), N-hydroxy-3-(2-(naphthalen-2-yl)thiazol-4-yl)propanamide (22), N -Hydroxy-4-(2-(naphthalen-2-yl)thiazol-4-yl)butanamide (23), N-hydroxy-3-(2-(6-fluoronaphthalen-2-yl) Thiazol-4-yl)-propanamide (24), N-hydroxy-3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)-propanamide (25), N -Hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide (26), N-hydroxy-3-(2-(quinolin-2-yl) Thiazol-4-yl)propanamide (27), N-hydroxy-3-(2-(quinolin-3-yl)thiazol-4-yl)propanamide (28), N-hydroxy-3 - (2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide (29), N-hydroxy-3-(2-(4'-fluoro-[ 1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide (30), N-hydroxy-3-(2-(4-phenoxyphenyl)thiazol-4-yl ) Propanamide (31), 3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide (32), (E)-N-hydride Roxy-3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanamide (33), (E)-N-hydroxy-3-(2-( 2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide (34), N-hydroxy-3-(2-(2-phenylcyclopropyl)thiazol-4-yl ) Propanamide (35), 3-(2-(2-(4-fluorophenyl)cyclopropyl)thiazol-4-yl)-N-hydroxypropanamide (36), N-hydroxy-3- (2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide (37), N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benz amide (38); and 4-fluoro-N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (39). , but is not limited to this.

The present invention provides a pharmaceutical composition for the prevention or treatment of HDAC6-mediated diseases comprising the above compound, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In another embodiment of the present invention, the composition may be characterized as selectively inhibiting HDAC6, but is not limited thereto.

In another embodiment of the present invention, the HDAC6-mediated disease includes organ transplant rejection, disease related to abnormal function of lymphocytes, cell proliferative disease, inflammatory disease, autosomal dominant disease, genetically related metabolic disease, autoimmune disease, It may be one or more selected from the group consisting of cancer disease, neurological disease, hypertrophy, heart failure, eye disease, or neurodegenerative disease, but is not limited thereto.

Additionally, the present invention provides a method for preventing or treating HDAC6-mediated diseases, comprising administering the composition.

Additionally, the present invention provides the use of a composition containing Compound 1 as an active ingredient for preventing or treating HDAC6-mediated diseases.

In addition, the present invention provides the use of a composition containing Compound 1 as an active ingredient for the production of a drug for treating HDAC6-mediated diseases.

According to the novel HDAC6 selective inhibitor and its use, the compound capable of selectively inhibiting HDAC6 has been optimized to present a new HDAC6 inhibitor mother structure, and not only has an excellent HAC6 inhibitory activity effect, but also inhibits HDAC6 compared to other lower HDACs. Since it can act selectively and effectively, it can be useful as a composition for preventing or treating HDAC6-mediated diseases.

Figure 1 is a diagram showing an in silico docking model of compound 26 with the HDAC6 X-ray crystal structure.
Figure 2 is a graph showing the results of molecular dynamics simulation analysis based on a docking model for compound 26.
Figure 3 is a graph showing the western blot results of compound 26 on HDAC6 inhibitory activity.
Figure 4 is a graph showing the western blot results of the inhibitory activity of Compound 26 on HDAC6 compared to SAHA.

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the following examples.

The compound of the present invention contains a structure capped with an ester group or acetamide group on one side of the thiazole group and a naphthalene cap group on the other side, and the double bond of the cap structure is expanded to form HDAC6. It produces a selective inhibitory effect. That is, according to the present invention, the double bond structure fixes the cap structure at a position showing residue differences between HDAC subtypes, and through this, it is confirmed that the double bond plays an important role in selectivity, resulting in a new core structure of the HDAC6 selective inhibitor. is presented. In addition, the compound of the present invention has excellent HDAC6 inhibitory activity and can be usefully used as a composition for preventing or treating HDAC6-mediated diseases.

The compound of the present invention is represented by the following formula (1) and includes isomers thereof or pharmaceutically acceptable salts thereof.

[Formula 1]

In the above formula, R is,

, , , , , , , , , , and is any one selected from the group consisting of, wherein X ₁ is any one selected from the group consisting of H, F, Br and OMe, where X ₂ is H or F, and where n is 0, 2, or 3 It is an integer.

In one embodiment of the present invention, the compound is N-hydroxy-3-(2-(naphthalen-1-yl)thiazol-4-yl)propanamide (20)(N-Hydroxy-3-( 2-(naphthalen-1-yl)thiazol-4-yl)propanamide (20)), N-hydroxy-2-(naphthalen-2-yl)thiazol-4-carboxamide (21)(N-Hydroxy -2-(naphthalen-2-yl)thiazole-4-carboxamide (21)), N-hydroxy-2-(2-(naphthalen-2-yl)thiazol-4-yl)acetamide (22)( N-Hydroxy-2-(2-(naphthalen-2-yl)thiazol-4-yl)acetamide(22)), N-hydroxy-4-(2-(naphthalen-2-yl)thiazol-4- 1) Butanamide (23) (N-Hydroxy-4-(2-(naphthalen-2-yl)thiazol-4-yl)butanamide (23)), N-hydroxy-3-(2-(6-fluo) Lonaphthalen-2-yl)thiazol-4-yl)-propanamide (24)(N-Hydroxy-3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)-propanamide (24) ), N-Hydroxy-3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)-propanamide (25) (N-Hydroxy-3-(2-(6-bromonaphthalen -2-yl)thiazol-4-yl)- propanamide (25)), N-hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide (26) )(N-Hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide (26)), N-hydroxy-3-(2-(quinolin-2-yl)thiazol Zol-4-yl)propanamide (27) (N-Hydroxy-3-(2-(quinolin-2-yl)thiazol-4-yl)propanamide (27)), N-hydroxy-3-(2- (quinolin-3-yl)thiazol-4-yl)propanamide (28) (N-hydroxy-3-(2-(quinolin-3-yl)thiazol-4-yl)propanamide (28)), N- Hydroxy-3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide (29)(N-Hydroxy-3-(2-([1, 1'-biphenyl]-4-yl)thiazol-4-yl)- propanamide (29)), N-hydroxy-3-(2-(4'-fluoro-[1,1'-biphenyl]- 4-yl)thiazol-4-yl)-propanamide (30)(N-Hydroxy-3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4- yl)- propanamide (30)), N-hydroxy-3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanamide (31)(N-Hydroxy-3-(2-(4) -phenoxyphenyl)thiazol-4-yl)propanamide (31)), 3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide (32) (3-(2-(4-(4-Fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide(32)), (E)-N-hydroxy-3-(2-(1-phenylpropanamide) Ph-1-en-2-yl)thiazol-4-yl)propanamide (33)((E)-N-Hydroxy-3-(2-(1-phenylprop-1-en-2-yl)thiazol -4-yl)propanamide (33)), (E)-N-hydroxy-3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide ( 34)((E)-N-Hydroxy-3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide(34)), N-hydroxy-3-(2 -(2-phenylcyclopropyl)thiazol-4-yl)propanamide (35)(N-Hydroxy-3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanamide (35)), 3-( 2-(2-(4-Fluorophenyl)cyclopropyl)thiazol-4-yl)-N-hydroxypropanamide (36)(3-(2-(2-(4-Fluorophenyl)cyclopropyl)thiazol- 4-yl)-N-hydroxypropanamide (36)), N-hydroxy-3-(2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide (37)(N-Hydroxy-3-( 2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide (37)), N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (38) (N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (38)); And 4-Fluoro-N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (39)(4-Fluoro-N-(4-(3- It may be any one selected from the group consisting of (hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (39)), but is not limited thereto.

The above compounds 20 to 39 can be synthesized from compounds 1 to 17, and the compounds 1 to 17 are methyl 3-(2-(naphthalen-1-yl)thiazol-4-yl)propanoate (1)( Methyl 3-(2-(naphthalen-1-yl)thiazol-4-yl)propanoate(1)), Ethyl 2-(naphthalen-2-yl)thiazole-4-carboxylate (2)(Ethyl 2- (naphthalen-2-yl)thiazole-4-carboxylate(2)), Methyl 2-(2-(naphthalen-2-yl)thiazol-4-yl)acetate (3)(Methyl 2-(2-(naphthalen -2-yl)thiazol-4-yl)acetate (3)), methyl 3-(2-(naphthalen-2-yl)thiazol-4-yl)propanoate (4)(Methyl 3-(2- (naphthalen-2-yl)thiazol-4-yl)propanoate (4)), methyl 4-(2-(naphthalen-2-yl)thiazol-4-yl)butanoate (5) (Methyl 4-( 2-(naphthalen-2-yl)thiazol-4-yl)butanoate (5)), methyl 3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)propanoate (6) )(Methyl 3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)propanoate(6)), Methyl 3-(2-(6-bromonaphthalen-2-yl)thiazol-4 -yl)propanoate (7) (Methyl 3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)propanoate(7)), methyl 3-(2-(6-methoxynaphthalene- 2-yl)thiazol-4-yl)propanoate (8) (Methyl 3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanoate(8)), Methyl 3-(2 -(Quinolin-2-yl)thiazol-4-yl)propanoate (9) (Methyl 3-(2-(quinolin-2-yl)thiazol-4-yl)propanoate (9)), Methyl 3- (2-(quinolin-3-yl)thiazol-4-yl)propanoate (10) (Methyl 3-(2-(quinolin-3-yl)thiazol-4-yl)propanoate (10)), methyl 3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate (11)(Methyl 3-(2-([1,1'-biphenyl]- 4-yl)thiazol-4-yl)propanoate(11)), methyl 3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl) Propanoate (12) (Methyl 3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate (12)), Methyl 3-(2- (4-phenoxyphenyl)thiazol-4-yl)propanoate (13) (Methyl 3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanoate (13)), Methyl 3-(2- (4-(4-fluorophenoxy)phenyl)thiazol-4-yl)propanoate (14)(Methyl 3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)propanoate (14)), (E)-methyl 3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanoate (15)((E)-Methyl 3 -(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanoate(15)), (E)-methyl 3-(2-(2-phenylprop-1-en- 1-yl)thiazol-4-yl)propanoate (16)((E)-Methyl 3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanoate(16) )), Methyl 3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanoate (17)(Methyl 3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanoate(17) )), but is not limited to this.

Compounds 1 to 17 are thiazole esters and can be prepared by methods common to those skilled in the art. In the examples of the present invention, thioamide was used, but is not limited thereto.

Compounds 20 to 39 of the present invention can be prepared from compounds 18 and 19, and compounds 18 and 19 are methyl 3- (2-benzamidothiazol-4-yl) propanoate (18) (Methyl 3- (2-benzamidothiazol-4-yl)propanoate(18)); And Methyl 3-(2-(4-fluorobenzamido)thiazol-4-yl)propanoate (19) (Methyl 3-(2-(4-fluorobenzamido)thiazol-4-yl)propanoate (19) ))am.

The compounds 18 and 19 may be prepared by amidating compounds 1 to 17, but are not limited thereto.

The process for amidating the compounds 1 to 17 can be applied without restrictions as long as it is a method common to those skilled in the art. In the examples of the present invention, hydrochloric acid and the like were used, but the process is not limited thereto.

Compounds 20 to 39 of the present invention may be hydroxamates prepared from compounds 18 and 19, but are not limited thereto.

The process for producing the hydroxamate can be applied without restrictions as long as it is a method common to those skilled in the art. In the examples of the present invention, methanol, thiazole ester, etc. were used, but it is not limited thereto.

The present invention provides a pharmaceutical composition for preventing or treating HDAC6-mediated diseases, comprising Compound 1 or a pharmaceutically acceptable salt thereof as an active ingredient.

In one embodiment of the present invention, the composition may selectively inhibit HDAC6, but is not limited thereto.

In one embodiment of the present invention, the HDAC6-mediated disease includes organ transplant rejection, disease related to abnormal function of lymphocytes, cell proliferative disease, inflammatory disease, autosomal dominant disease, genetically related metabolic disease, autoimmune disease, and cancer. It may be one or more selected from the group consisting of disease, neurological disease, hypertrophy, heart failure, eye disease, or neurodegenerative disease, but is not limited thereto.

The disease according to the present invention is a disease related to HDAC6, meaning that it is mediated by HDAC6, for example, Huntington's disease (HD), Alzheimer's disease (AD), Parkinson's disease (PD), and amyotrophic axon. ALS may be present, but is not limited thereto.

The pharmaceutical composition for prevention or treatment according to the present invention may further include appropriate carriers, excipients, and diluents commonly used in the preparation of pharmaceutical compositions. The excipient may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, humectants, film-coating materials, and controlled-release additives.

The pharmaceutical composition according to the present invention can be prepared as powder, granules, sustained-release granules, enteric-coated granules, solutions, eye drops, ellipsis, emulsions, suspensions, spirits, troches, perfumes, and limonadese according to conventional methods. , tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric-coated capsules, pills, tinctures, soft extracts, dry extracts, liquid extracts, injections, capsules, perfusate, It can be formulated and used in the form of external preparations such as warning agents, lotions, pasta preparations, sprays, inhalants, patches, sterilized injection solutions, or aerosols, and the external preparations include creams, gels, patches, sprays, ointments, and warning agents. , it may have a dosage form such as lotion, liniment, pasta, or cataplasma.

Carriers, excipients, and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, and calcium. These include phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Additives to tablets, powders, granules, capsules, pills, and troches according to the present invention include corn starch, potato starch, wheat starch, lactose, white sugar, glucose, fructose, di-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, and phosphoric acid. Calcium monohydrogen, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropylmethyl. Excipients such as cellulose (HPMC), HPMC 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethyl cellulose, calcium carboxymethyl cellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethyl cellulose, sodium methyl cellulose, methyl cellulose, microcrystalline cellulose, dextrin. , hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, etc. binders can be used, Hydroxypropyl methyl cellulose, corn starch, agar powder, methyl cellulose, bentonite, hydroxypropyl starch, sodium carboxymethyl cellulose, sodium alginate, calcium carboxymethyl cellulose, calcium citrate, sodium lauryl sulfate, silicic acid anhydride, 1-hydroxy Propylcellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, sodium bicarbonate, polyvinylpyrrolidone, calcium phosphate, gelled starch, gum arabic, Disintegrants such as amylopectin, pectin, sodium polyphosphate, ethyl cellulose, white sugar, magnesium aluminum silicate, di-sorbitol solution, light anhydrous silicic acid; Calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium, kaolin, petrolatum, sodium stearate, cacao fat, sodium salicylate, magnesium salicylate, polyethylene glycol (PEG) 4000, PEG 6000, liquid paraffin, hydrogen. Added soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, Macrogol, synthetic aluminum silicate, silicic anhydride, higher fatty acids, higher alcohol, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, Lubricants such as starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid may be used.

Additives for the liquid according to the present invention include water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, sucrose monostearate, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, Lanolin esters, acetic acid, hydrochloric acid, aqueous ammonia, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

A solution of white sugar, other sugars, or sweeteners, etc. may be used in the syrup according to the present invention, and if necessary, flavoring agents, colorants, preservatives, stabilizers, suspending agents, emulsifiers, thickening agents, etc. may be used.

Purified water can be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. can be used as needed.

Suspensions according to the present invention include acacia, tragacantha, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, HPMC 2910, etc. Topics may be used, and surfactants, preservatives, stabilizers, colorants, and fragrances may be used as needed.

Injections according to the present invention include distilled water for injection, 0.9% sodium chloride injection, IV solution, dextrose injection, dextrose + sodium chloride injection, PEG, lactated IV solution, ethanol, propylene glycol, non-volatile oil - sesame oil. , solvents such as cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristic acid, and benzene benzoate; Solubilizers such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, Tween, nicotinic acid amide, hexamine, and dimethylacetamide; Weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and buffering agents such as gums; Isotonic agents such as sodium chloride; Stabilizers such as sodium bisulfite (NaHSO3) carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; Sulfurizing agents such as sodium bisulfide 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetate, and acetone sodium bisulfite; Analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; It may contain suspending agents such as CM sodium, sodium alginate, Tween 80, and aluminum monostearate.

Suppositories according to the present invention include cacao oil, lanolin, witepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cacao butter + Cholesterol, lecithin, Lanet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego -G), Cebes Pharma 16, Hexalide Base 95, Cotomar, Hydrocote SP, S-70-XXA, S-70-XX75(S-70-XX95), Hydro Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Massaupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), suppositories type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecobi (W, R, S, M, Fs), Tegestor triglyceride base (TG-95, MA, 57) and The same mechanism can be used.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the extract with at least one excipient, such as starch, calcium carbonate, and sucrose. ) or prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium styrate talc are also used.

Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate.

The present invention provides a method for preventing or treating HDAC6-mediated diseases, comprising administering the composition.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat the disease with a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is determined by the type, severity, drug activity, and It can be determined based on factors including sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other factors well known in the medical field.

The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiple times. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art to which the present invention pertains.

The pharmaceutical composition of the present invention can be administered to an individual through various routes. All modes of administration are contemplated, including oral administration, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, paraspinal space (intrathecal) injection, sublingual administration, buccal administration, intrarectal injection, vaginal injection. It can be administered by internal insertion, ocular administration, ear administration, nasal administration, inhalation, spraying through the mouth or nose, dermal administration, transdermal administration, etc.

The pharmaceutical composition of the present invention is determined depending on the type of drug as the active ingredient along with various related factors such as the disease to be treated, the route of administration, the patient's age, gender, weight, and severity of the disease.

In the present invention, “individual” refers to a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, rats, dogs, cats, horses, cows, etc. refers to mammals of

In the present invention, “administration” means providing a given composition of the present invention to an individual by any suitable method.

In the present invention, “prevention” refers to any action that suppresses or delays the onset of the desired disease, and “treatment” refers to the improvement or improvement of the desired disease and its associated metabolic abnormalities by administration of the pharmaceutical composition according to the present invention. It refers to all actions that are beneficially changed, and “improvement” refers to all actions that reduce parameters related to the desired disease, such as the degree of symptoms, by administering the composition according to the present invention.

“Histone deacetylases (HDACs)” according to the present invention can be attached to gene promoters by corepressors or multi-protein transcriptional complexes, where they do not bind directly to DNA but There are 18 encoded human HDACs, which are enzymes that function to regulate transcription through chromatin modification, divided into class I (HDACs 1, 2, 3, and 8) and class II (HDACs 4, 5, and 6). , 7, 9, and 10), class III (SIRT 1-7), and class IV (HDAC11) enzymes.

“HDAC6” according to the present invention is a member of the class IIb family of HDACs and acts as a cytoplasmic deacetylase associated with microtubules (MTs).

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by the following examples.

[Example]

Example 1. Preparation of compounds 1 to 17

To prepare compounds 1 to 17, thiazole esters were prepared according to general manufacturing procedures. Specifically, bromo ester (1.2 equiv, 1.2 equi) was added to a solution of the appropriate thioamide (1 equi, 1 equiv) in methanol or ethanol (0.16 mmol mL ^-1 ) and refluxed for 16 hours. After completion of the reaction, the solvent was removed under reduced pressure, the residue was diluted with ethyl acetate and washed with water and brine, and the organic layer was dried over MgSO ₄ and concentrated. The crude product was purified by flash chromatography on silica gel (hexane-ethyl acetate) to obtain the thiazole ester (Table 1).

division designation structure Compound 1 Methyl 3-(2-(naphthalen-1-yl)thiazol-4-yl)propanoate compound 2 Ethyl 2-(naphthalen-2-yl)thiazole-4-carboxylate Compound 3 Methyl 2-(2-(naphthalen-2-yl)thiazol-4-yl)acetate Compound 4 Methyl 3-(2-(naphthalen-2-yl)thiazol-4-yl)propanoate Compound 5 Methyl 4-(2-(naphthalen-2-yl)thiazol-4-yl)butanoate Compound 6 Methyl 3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)propanoate Compound 7 Methyl 3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)propanoate Compound 8 Methyl 3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanoate Compound 9 Methyl 3-(2-(quinolin-2-yl)thiazol-4-yl)propanoate Compound 10 Methyl 3-(2-(quinolin-3-yl)thiazol-4-yl)propanoate Compound 11 Methyl 3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate Compound 12 Methyl 3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate Compound 13 Methyl 3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanoate Compound 14 Methyl 3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)propanoate Compound 15 (E)-Methyl 3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanoate Compound 16 (E)-Methyl 3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanoate Compound 17 Methyl 3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanoate

Compound 1:Methyl 3-(2-(naphthalen-1-yl)thiazol-4-yl)propanoate

White solid in 49% yield. 1H NMR (400 MHz, CDCl3) δ 8.74-8.76 (d, J = 8.4 Hz, 1H), 7.87 -7.93 (m, 2H), 7.77-7.79 (dd, J = 7.2, 0.8 Hz, 1H), 7.48- 7.59 (m, 3H), 7.07 (s, 1H), 3.70 (s, 3H), 3.22-3.26 (t, J = 7.6 Hz, 2H), 2.86-2.90 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.4, 167.2, 156.1, 134.0, 130.9, 130.5, 130.3, 128.4, 128.3, 127.3, 126.3, 125.9, 125.0, 114.5, 51.7, 33.6, 26.9.

Compound 2: Ethyl 2-(naphthalen-2-yl)thiazole-4-carboxylate

In this reaction, ethyl 3-bromo-2-oxopropanoate was used as a starting material and ethanol as a solvent. Colorless solid in 18% yield. 1H NMR (400 MHz, CDCl3) δ 8.50 (s, 1H), 8.18 (s, 1H), 8.08-8.11 (dd, J = 2.0, 8.4 Hz, 1H), 7.84-7.94 (m, 3H), 7.51- 7.54 (m, 2H), 4.44-4.49 (q, J = 7.2 Hz, 2H), 1.43-1.46 (t, J = 7.0 Hz, 3H); 13C NMR (100 MHz, CDCl3) δ: 168.9, 161.4, 148.2, 134.3, 133.1, 130.1, 128.8, 128.7, 127.8, 127.3, 127.0, 126.9, 126.7, 124.1, 61.5, 14.3.

Compound 3: Methyl 2-(2-(naphthalen-2-yl)thiazol-4-yl)acetate

White solid in 34% yield. 1H NMR (400 MHz, CDCl3) δ 8.41 (s, 1H), 8.01-8.04 (dd, J = 1.8, 8.6 Hz, 1H), 7.82-7.91 (m, 3H), 7.48-7.52 (m, 2H), 7.23 (s, 1H), 3.94 (s, 2H), 3.76 (s, 3H); 13C NMR (100 MHz, CDCl3) δ 170.8, 168.0, 149.8, 134.1, 133.2, 130.8, 128.7, 128.6, 127.8, 126.9, 126.7, 125.9, 124.0, 116.2, 52.2, 37.0.

Compound 4: Methyl 3-(2-(naphthalen-2-yl)thiazol-4-yl)propanoate

Yellow oil in 76% yield. 1H NMR (400 MHz, CDCl3) δ 8.40 (s, 1H), 8.01-8.04 (dd, J = 2.0, 8.4 Hz, 1H), 7.82-7.91 (m, 3H), 7.48-7.52 (m, 2H), 6.98 (s, 1H), 3.70 (s, 3H), 3.16-3.20 (t, J = 7.2 Hz, 2H), 2.83-2.87 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.2, 167.8, 156.4, 133.9, 133.2, 131.0, 128.6, 128.5, 127.7, 126.8, 126.6, 125.7, 123.9, 113.7, 51.6, 33.4, 26.8.

Compound 5: Methyl 4-(2-(naphthalen-2-yl)thiazol-4-yl)butanoate

White solid in 22% yield. 1H NMR (400 MHz, CDCl3) δ 8.40 (s, 1H), 8.02-8.04 (dd, J = 1.8, 8.6 Hz, 1H), 7.81-7.91 (m, 3H), 7.46-7.51 (m, 2H), 6.92 (s, 1H), 3.67 (s, 3H), 2.87-2.91 (t, J = 7.2 Hz, 2H), 2.41-2.45 (t, J = 7.6 Hz, 2H), 2.10-2.17 (m, 2H) ; 13C NMR (100 MHz, CDCl3) δ 173.8, 167.8, 157.5, 134.0, 133.3, 128.6, 128.6, 127.8, 126.8, 126.7, 125.7, 124.1, 113.5, 51.5, 33.4, 3 0.9, 24.4.

Compound 6: Methyl 3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)propanoate

White solid in 74% yield. 1H NMR (400 MHz, CDCl3) δ 8.38 (s, 1H), 8.02-8.05 (d, J = 8.4 Hz, 1H), 7.87-7.90 (dd, J = 5.6, 8.8 Hz, 1H), 7.79-7.81 ( d, J = 8.4 Hz, 1H), 7.43-7.46 (dd, J = 2.4, 9.6 Hz, 1H), 7.25-7.30 (m, 1H), 6.98 (s, 1H), 3.70 (s, 3H), 3.16 -3.19 (t, J = 7.6 Hz, 2H), 2.83-2.86 (t, J = 7.2 Hz, 2H) ); 13C NMR (100 MHz, CDCl3) δ 173.3, 167.5, 162.4, 159.9, 156.5, 134.8, 134.7, 131.0, 130.9, 130.6, 130.5, 130.2, 125.6, 125.1, 117.3, 117.0, 113.8, 111.1, 110.9, 51.7, 33.5 , 26.8.

Compound 7: Methyl 3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)propanoate

White solid in 22% yield. 1H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.99-8.02 (dd, J = 1.6 Hz, 1H), 7.95 (d, J = 1.6 Hz, 1H), 7.73-7.74 (d, J = 4.4 Hz, 1H), 7.70-7.72 (d, J = 4.8 Hz, 1H), 7.52-7.55 (dd, J = 2.0, 8.4 Hz, 1H), 6.97 (s, 1H), 3.70 (s, 3H), 3.15-3.18 (t, J = 7.6 Hz, 2H), 2.81-2.85 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.2, 167.2, 156.7, 134.9, 131.6, 131.52, 130.1, 129.9, 127.7, 125.5, 125.1, 120.9, 114.0, 51.7, 33.4, 26.8.

Compound 8: Methyl 3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanoate

White solid in 52% yield. 1H NMR (400 MHz, CDCl3) δ 8.33 (s, 1H), 7.98-8.00 (dd, J = 1.6, 8.4 Hz, 1H), 7.79-7.82 (d, J = 8.8 Hz, 1H), 7.76-7.78 ( d, J = 8,8 Hz, 1H), 7.14-7.26 (m, 2H), 6.95 (s, 1H), 3.94 (s, 3H), 3.71 (s, 3H), 3.15-3.19 (t, J = 7.6 Hz, 2H), 2.83-2.86 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ: 173.3, 168.1, 158.5, 156.3, 135.4, 130.1, 129.1, 128.6, 127.4, 125.6, 124.6, 119.6, 113.2, 105.8, 55.3, 51.6, 33.5, 26.9.

Compound 9: Methyl 3-(2-(quinolin-2-yl)thiazol-4-yl)propanoate

White solid in 42% yield. 1H NMR (400 MHz, CDCl3) δ 8.28-8.30 (d, J = 8.8 Hz, 1H), 8.21-8.23 (d, J = 8.4 Hz, 1H), 8.10-8.13 (d, J = 8.8 Hz, 1H) , 7.81-7.83 (dd, J = 1.2, 8.4 Hz, 1H), 7.70-7.74 (m, 1H), 7.52-7.56 (m, 1H), 7.12 (s, 1H), 3.70 (s, 3H), 3.17 -3.21 (t, J = 7.6 Hz, 2H), 2.83-2.87 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDClc) δ 173.32, 169.01, 156.82, 151.34, 147.86, 136.90, 129.92, 129.50, 128.59, 127.68, 127.01, 117.88, 117.00, 51.70, 33.54, 26.94.

Compound 10: Methyl 3-(2-(quinolin-3-yl)thiazol-4-yl)propanoate

Colorless oil in 44% yield. 1H NMR (400 MHz, CDCl3) δ 9.44-9.45 (d, J = 2.0 Hz, 1H), 8.64 (d, J = 2 Hz, 1H), 8.12-8.14 (d, J = 8.0 Hz, 1H), 7.89 -7.91 (dd, J = 1.2, 8.4 Hz, 1H), 7.73-7.77 (m, 1H), 7.57-7.61 (m, 1H), 7.06 (s, 1H), 3.71 (s, 3H), 3.18-3.22 (t, J = 7.6 Hz, 2H), 2.84-2.88 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.2, 164.5, 157.0, 148.5, 132.9, 130.3, 129.4, 128.4, 127.5, 127.4, 126.8, 114.5, 51.7, 33.4, 26.7.

Compound 11: Methyl 3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate

White solid in 30% yield. 1H NMR (400 MHz, CDCl3) δ 7.98-8.01 (m, 2H), 7.62-7.67 (m, 2H), 7.44-7.48 (m, 2H), 7.35-7.39 (m, 2H), 6.96 (s, 1H) ), 3.70 (s, 3H), 3.14-3.18 (t, J = 7.6 Hz, 2H), 2.81-2.85 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.3, 167.5, 156.4, 142.6, 140.2, 132.6, 128.8, 127.7, 127.5, 127.0, 126.9, 113.6, 51.7, 33.5, 26.8.

Compound 12: Methyl 3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)propanoate

White solid in 38% yield. 1H NMR (400 MHz, CDCl3) δ 7.97-7.99 (d, J = 8.4 Hz, 2H), 7.56-7.60 (m, 4H), 7.12-7.16 (t, J = 8.8 Hz, 2H), 6.96 (s, 1H), 3.70 (s, 3H), 3.14-3.18 (t, J = 7.2 Hz, 2H), 2.81-2.85 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.3, 167.3, 163.9, 161.4, 156.5, 141.5, 136.3, 132.7, 128.6, 128.6, 127.3, 126.9, 115.9, 115.6, 113.6, 51.7, 33.5, 26.8.

Compound 13: Methyl 3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanoate

White solid in 29% yield. 1H NMR (400 MHz, CDCl3) δ 7.81 - 7.94 (m, 2H), 7.34 - 7.42 (m, 2H), 7.12 - 7.20 (m, 1H), 6.98 - 7.10 (m, 4H), 6.92 (s, 1H) ), 3.70 (s, 3H), 3.14 (t, J = 7.46 Hz, 2H), 2.82 (t, J = 7.46 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.3, 167.2, 159.0, 156.4, 156.2, 129.9, 128.8, 128.1, 123.8, 119.4, 118.7, 113.2, 51.7, 33.5, 29.7, 26 .8.

Compound 14: Methyl 3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)propanoate

Yellow oil in 77% yield; 1H NMR (400 MHz, CDCl3) δ 7.88 (s, 1H), 7.86 (s, 1H), 7.06-7.01 (m, 4H), 7.00 (s, 1H), 6.97 (s, 1H), 6.91 (s, 1H), 3.69 (s, 3H), 3.15-3.11 (t, J = 7.6 Hz, 2H), 2.82-2.79 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.3, 167.1, 159.2, 156.2, 140.1, 131.0, 128.8, 128.1, 121.0, 118.1, 116.6, 116.3, 113.2, 51.7, 33.4, 2 6.8.

Compound 15: (E)-Methyl 3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanoate

Colorless oil in 23% yield. 1H NMR (400 MHz, CDCl3) δ 7.47 (s, 1H), 7.37 - 7.45 (m, 4H), 7.30 (d, J = 6.72 Hz, 1H), 6.89 (s, 1H), 3.71 (s, 3H) , 3.13 (t, J = 7.46 Hz, 2H), 2.80 (t, J = 7.52 Hz, 2H), 2.34 - 2.43 (m, 3H).

Compound 16: (E)-Methyl 3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanoate

Colorless oil in 46% yield. 1H NMR (400 MHz, DMSO-d6) δ 7.59 - 7.63 (m, 2H), 7.34 - 7.44 (m, 4H), 7.04 (dd, J = 0.49, 1.22 Hz, 1H), 3.61 (s, 3H), 3.01 (t, J = 7.46 Hz, 2H), 2.75 (t, J = 7.34 Hz, 2H), 2.55 (d, J = 1.22 Hz, 3H); 13C NMR (100 MHz, DMSO-d6) δ 172.6, 164.4, 155.6, 142.2, 141.4, 128.5, 128.1, 126.0, 119.5, 114.4, 51.4, 32.7, 26.2, 18.2.

Compound 17: Methyl 3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanoate

Colorless oil in 64% yield. 1H NMR (400 MHz, CDCl3) δ 7.30 (t, J = 7.46 Hz, 2H), 7.21 (t, J = 7.21 Hz, 1H), 7.15 (d, J = 7.82 Hz, 2H), 6.72 (s, 1H) ), 3.70 (s, 3H), 3.05 (t, J = 7.52 Hz, 2H), 2.75 (t, J = 7.52 Hz, 2H), 2.48 - 2.58 (m, 2H), 1.70 - 1.79 (m, 1H) , 1.53 - 1.61 (m, 1H)

Example 2. Preparation of Compounds 18 and 19

To prepare compounds 18 and 19, amidation was performed according to general manufacturing procedures. Specifically, acid (2.68 mmol), EDCI·HCl (5.36 mmol), and HOBt hydrate (5.36 mmol) were dissolved in dichloromethane (5.36 mL). Afterwards, diisopropylethylamine (8.04 mmol) and amine (3.22 mmol) were added to the mixture and stirred at room temperature overnight. The reaction was quenched with 1N HCl and extracted with dichloromethane, and the extract was washed with water, brine, dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by flash silica gel column chromatography to obtain compounds 18 and 19 (Table 2).

division designation structure Compound 18 Methyl 3-(2-benzamidotiazol-4-yl)propanoate Compound 19 Methyl 3-(2-(4-fluorobenzamido)thiazol-4-yl)propanoate

Compound 18: Methyl 3-(2-benzamidotiazol-4-yl)propanoate

Yellow-white solid in 94% yield. 1H NMR (400 MHz, CDCl3) δ 7.90-7.92 (m, 2H), 7.55-7.59 (m, 1H), 7.25-7.29 (m, 2H), 6.60 (s, 1H), 3.63 (s, 3H), 2.76-2.79 (t, J = 7.6 Hz, 2H), 2.57-2.60 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.0, 164.9, 158.6, 149.8, 132.8, 132.2, 128.9, 127.5, 108.4, 51.6, 33.1, 26.4.

Compound 19: Methyl 3-(2-(4-fluorobenzamido)thiazol-4-yl)propanoate

Yellow-white solid in 92% yield. 1H NMR (400 MHz, CDCl3) δ 7.92-7.95 (m, 2H), 7.14-7.18 (t, J = 8.4 Hz, 2H), 6.63 (s, 1H), 3.64 (s, 3H), 2.74-2.78 ( t, J = 7.6 Hz, 2H), 2.57-2.60 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, CDCl3) δ 173.0, 166.7, 164.2, 163.9, 158.7, 149.7, 130.1, 130.1, 128.4, 128.5, 116.2, 116.0, 108.6, 51.7, 33.1, 2 6.3.

Example 3. Preparation of compounds 20 to 39

To prepare compounds 20 to 39, hydroxamates were prepared according to general manufacturing procedures. Specifically, hydroxylamine chloride (2equ) and 8N sodium hydroxide (0.5 mmol mL ^-1 ) were added to a solution of an appropriate thiazole ester (1equ) in methanol (0.1 mmol mL ^-1 ) at 0°C. The reaction mixture was stirred at room temperature for 1.5 hours to complete the reaction, and then methanol was removed under reduced pressure. The residue was diluted with water and extracted with ethyl acetate. The complex extract was washed with water and brine, dried over MgSO ₄ and concentrated. The crude product was recrystallized from ethyl acetate or methylene chloride to give compounds 20 to 39 (Table 3).

division designation structure Compound 20 N-Hydroxy-3-(2-(naphthalen-1-yl)thiazol-4-yl)propanamide Compound 21 N-Hydroxy-2-(naphthalen-2-yl)thiazole-4-carboxamide Compound 22 N-Hydroxy-3-(2-(naphthalen-2-yl)thiazol-4-yl)propanamide Compound 23 N-Hydroxy-4-(2-(naphthalen-2-yl)thiazol-4-yl)butanamide Compound 24 N-Hydroxy-3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)-propanamide Compound 25 N-Hydroxy-3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)-propanamide Compound 26 N-Hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide Compound 27 N-Hydroxy-3-(2-(quinolin-2-yl)thiazol-4-yl)propanamide Compound 28 N-hydroxy-3-(2-(quinolin-3-yl)thiazol-4-yl)propanamide Compound 29 N-Hydroxy-3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)- propanamide Compound 30 N-Hydroxy-3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide Compound 31 N-Hydroxy-3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanamide Compound 32 3-(2-(4-(4-Fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide Compound 33 (E)-N-Hydroxy-3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanamide Compound 34 (E)-N-Hydroxy-3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide Compound 35 N-Hydroxy-3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanamide Compound 36 3-(2-(2-(4-Fluorophenyl)cyclopropyl)thiazol-4-yl)-N-hydroxypropanamide Compound 37 N-Hydroxy-3-(2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide Compound 38 N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide Compound 39 4-Fluoro-N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide

Compound 20: N-Hydroxy-3-(2-(naphthalen-1-yl)thiazol-4-yl)propanamide

White solid in 19% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.51 (br. s., 1H), 8.84 (d, J = 8.07 Hz, 1H), 8.79 (br. s., 1H), 8.00 - 8.10 (m, 2H) ), 7.87 (d, J = 7.09 Hz, 1H), 7.58 - 7.67 (m, 3H), 7.48 (s, 1H), 3.09 (t, J = 7.58 Hz, 2H), 2.47 (m, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.7, 166.4, 157.1, 134.1, 130.8, 130.6, 130.1, 128.9, 128.8, 127.9, 127.0, 126.1, 125.9, 115.7, 32.3 , 27.5.

Compound 21: N-Hydroxy-2-(naphthalen-2-yl)thiazole-4-carboxamide

White solid in 20% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.58 (s, 1H), 8.55 (s, 1H), 8.04-8.11 (m. 3H), 7.96-7.98 (m, 1H), 7.58-7.60 (m, 2H) ); 13C NMR (100 MHz, DMSO-d6) δ 167.93, 162.57, 148.73, 134.28, 133.28, 130.41, 129.47, 129.44, 129.17, 128.25, 127.99, 127.58, 126.53 , 124.02.

Compound 22: N-Hydroxy-3-(2-(naphthalen-2-yl)thiazol-4-yl)propanamide

White solid in 11% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.00-8.10 (m, 3H), 7.95-7.96 (m, 1H), 7.56-7.58 (m, 2H), 7.40 (s, 1H) ), 3.03-3.04 (t, J = 7.2 Hz, 2H), 2.43-2.47 (t, J = 7.2 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.6, 166.9, 157.8, 134.0, 133.3, 131.0, 129.2, 129.0, 128.2, 127.6, 127.4, 125.6, 12.0, 115.2, 32.3, 27.4.

Compound 23: N-Hydroxy-4-(2-(naphthalen-2-yl)thiazol-4-yl)butanamide

White solid in 11% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.49 (s, 1H), 7.93-8.09 (m, 4H), 7.55-7.59 (m, 2H), 7.39 (s, 1H), 2.76-2.80 (t, J = 7.6 Hz, 2H), 2.05-2.09 (t, J = 7.2 Hz, 2H), 1.92-2.00 (m, 2H); 13C NMR (100 MHz, DMSO-d6) δ 169.3, 166.8, 158.0, 134.0, 133.3, 131.1, 129.2, 129.0, 128.2, 127.5, 127.4, 125.6, 124.1, 115.1, 32.3 , 30.9, 25.3.

Compound 24: N-Hydroxy-3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)-propanamide

White solid in 33% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.48 (s, 1H), 8.77 (s, 1H), 8.54 (s, 1H), 8.16-8.19 (m, 1H), 8.08-8.10 (d, J = 8.4 Hz, 1H) 7.99-8.01 (d, J = 8.0 Hz, 1H), 7.75-7.77 (d, J = 10.0 Hz, 1H), 7.47-7.51 (m, 1H), 7.39 (s, 1H), 3.01- 3.04 (t, J = 6.8 Hz, 2H), 2.43-2.46 (t, J = 6.4 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.6, 166.6, 162.3, 159.9, 157.3, 134.9, 134.8, 132.1, 132.0, 130.6, 128.7, 128.7, 125.7, 125.1, 117. 7, 117.5, 115.2, 111.5, 111.3, 32.2 , 27.4.

Compound 25: N-Hydroxy-3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)-propanamide

White solid in 10% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.48 (br. s., 1H), 8.77 (br. s, 1H), 8.53 (s, 1H), 8.27 (s, 1H), 8.09 - 8.13 (m, 1H), 8.06 (d, J = 8.80 Hz, 1H), 7.98 - 8.04 (m, 1H), 7.70 (dd, J = 1.77, 8.74 Hz, 1H), 7.42 (s, 1H), 3.02 (t, J = 7.58 Hz, 2H), 2.44 (t, J = 7.64 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.2, 166.0, 157.0, 134.6, 131.5, 131.1, 130.8, 130.0, 129.7, 128.1, 125.2, 124.8, 120.4, 115.1, 31.7 , 26.9.

Compound 26: N-Hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide

White solid in 18% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.41 (s, 1H), 7.97-8.01 (m, 2H), 7.88-7.91 (d, J = 8.8 Hz, 1H), 7.37 (d, J = 2 Hz, 1H), 7.32 (d, J = 2.4 Hz, 1H), 7.20-7.23 (dd, J = 2.4, 8.8 Hz, 1H), 3.90 (s, 1H), 2.97-3.01 (t, J = 7.6 Hz, 2H) ), 2.37-2.41 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.1, 167.0, 158.6, 157.7, 135.5, 130.6, 129.0, 128.7, 128.0, 125.5, 124.5, 119.9, 114.3, 106.5, 55.7 , 32.6, 27.9.

Compound 27: N-Hydroxy-3-(2-(quinolin-2-yl)thiazol-4-yl)propanamide

White solid in 10% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.49 (br. s., 1H), 8.79 (s, 1H), 8.52-8.54 (d, J = 8.8 Hz, 1H), 8.26-8.28 (d, J = 8.4 Hz, 1H), 8.02-8.06 (t, J = 7.6 Hz, 2H), 7.79-7.83 (m, 1H), 7.63-7.66 (t, J = 7.6 Hz, 1H), 7.52 (s, 1H), 3.03-3.07 (t, J = 7.6 Hz, 2H), 2.44-2.48 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, DMSO-D6) δ 168.5, 168.2, 157.8, 151.1, 147.6, 145.5, 138.1, 131.1, 129.1, 128.7, 128.6, 127.7, 117.9, 31.9, 27.4.

Compound 28: N-hydroxy-3-(2-(quinolin-3-yl)thiazol-4-yl)propanamide

White solid in 19% yield. 1H NMR (400 MHz, DMSO-d6) 1H NMR (400 MHz, DMSO-d6) δ 10.49 (s, 1H), 9.44 (d, J = 2.08 Hz, 1H), 8.88 (d, J = 1.96 Hz, 1H ), 8.78 (s, 1H), 8.15 (d, J = 8.07 Hz, 1H), 8.07 (d, J = 8.44 Hz, 1H), 7.83 (dt, J = 1.28, 7.67 Hz, 1H), 7.64 - 7.72 (m, 1H), 7.49 (s, 1H), 3.05 (t, J = 7.52 Hz, 2H), 2.39 - 2.48 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.1, 163.6, 157.1, 147.9, 147.8, 132.8, 130.6, 128.8, 127.6, 127.3, 126.2, 115.7, 31.7, 26.8

Compound 29: N-Hydroxy-3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide

White solid in 18% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.47 (s, 1H), 8.76 (s, 1H), 8.00-8.01 (d, J = 7.6 Hz, 2H), 7.79-7.81 (d, J = 8.0 Hz, 2H), 7.73-7.75 (d, J = 7.6 Hz, 2H), 7.48-7.52 (t, J = 7.6 Hz, 2H), 7.41-7.43 (d, J = 7.2 Hz, 1H), 7.36 (s, 1H) ), 2.99-3.03 (t, J = 7.6 Hz, 2H), 2.41-2.45 (t, J = 7.6 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.6, 166.4, 157.3, 142.0, 139.5, 132.6, 129.5, 128.4, 127.8, 127.1, 127.0, 115.0, 32.1, 27.3.

Compound 30: N-Hydroxy-3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide

White solid in 18% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.51 (br. s., 1H), 8.80 (br. s., 1H), 7.98 (d, J = 7.70 Hz, 2H), 7.76 (d, J = 6.60) Hz, 4H), 7.21 - 7.42 (m, 3H), 3.01 (t, J = 6.85 Hz, 2H), 2.44 (t, J = 7.03 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.2, 165.9, 163.4, 160.9, 156.9, 140.5, 135.6, 135.6, 132.2, 128.7, 128.7, 127.3, 126.6, 116.0, 115. 8, 114.6, 31.7, 26.9.

Compound 31: N-Hydroxy-3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanamide

White solid in 18% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.45 (s, 1H), 8.74 (s, 1H), 7.91-7.93 (d, J = 8.4 Hz, 2H), 7.42-7.46 (t, J = 7.6 Hz, 2H), 7.29 (s, 1H), 7.19-7.23 (t, J = 7.6 Hz, 1H), 7.07-7.11 (m, 4H), 2.95-2.99 (t, J = 7.6 Hz, 2H), 2.38-2.42 (t, J = 7.2 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.6, 166.2, 158.9, 157.0, 156.2, 130.7, 128.8, 128.4, 124.7, 119.8, 118.9, 114.5, 32.1, 27.3.

Compound 32: 3-(2-(4-(4-Fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide

White solid in 7% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.51 (br. s., 1H), 8.97 (br. s., 1H), 7.89 - 7.93 (m, 2H), 7.25 - 7.31 (m, 3H), 7.14 - 7.18 (m, 2H), 7.03 - 7.07 (m, 2H), 2.97 (t, J = 7.64 Hz, 2H), 2.40 (t, J = 7.64 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.1, 165.8, 159.9, 158.8, 157.5, 156.6, 151.7, 151.7, 128.2, 128.0, 121.6, 121.5, 118.0, 116.9, 116. 7, 114.1, 31.7, 26.9

Compound 33: (E)-N-Hydroxy-3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanamide

White solid in 43% yield. 1H NMR (500 MHz, DMSO-d6) δ 7.49 (d, J = 7.33 Hz, 2H), 7.38 - 7.46 (m, 3H), 7.30 - 7.35 (m, 1H), 7.28 (s, 1H), 2.96 ( t, J = 7.48 Hz, 2H), 2.64 (t, J = 7.48 Hz, 2H), 2.34 (d, J = 1.17 Hz, 3H); 13C NMR (125 MHz, DMSO-d6) δ 173.7, 169.8, 156.0, 136.1, 130.8, 129.7, 129.4, 128.4, 127.6, 113.8, 33.2, 26.5, 16.4.

Compound 34: (E)-N-Hydroxy-3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide

White solid in 26% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.45 (s, 1H), 8.74 (s, 1H), 7.62 (d, J = 7.21 Hz, 2H), 7.39 - 7.44 (m, 2H), 7.36 (d, J = 7.09 Hz, 1H), 7.33 (s, 1H), 7.05 (s, 1H), 2.97 (t, J = 7.64 Hz, 2H), 2.56 (d, J = 0.98 Hz, 3H), 2.35 - 2.43 ( m, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.2, 164.3, 156.1, 142.2, 141.4, 128.6, 128.1, 126.0, 119.7, 114.4, 31.8, 26.9, 18.3.

Compound 35: N-Hydroxy-3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanamide

Yellow solid in 23% yield. 1H NMR (400 MHz, DMSO-d6) δ 10.44 (br. s., 1H), 8.76 (br. s., 1H), 7.25 - 7.32 (m, 2H), 7.17 - 7.22 (m, 3H), 7.04 (s, 1H), 2.86 (t, J = 7.58 Hz, 2H), 2.64 - 2.70 (m, 1H), 2.44 - 2.50 (m, 1H), 2.32 (t, J = 7.52 Hz, 2H), 1.55 - 1.68 (m, 2H); 13C NMR (100 MHz, DMSO-d6) δ 170.1, 168.2, 162.0, 159.6, 155.1, 137.0, 137.0, 127.6, 127.5, 115.2, 115.0, 111.8, 31.7, 28.0, 26.9, 25.3, 19.8.

Compound 36: 3-(2-(2-(4-Fluorophenyl)cyclopropyl)thiazol-4-yl)-N-hydroxypropanamide

Yellow solid in 19% yield. 1H NMR (500 MHz, DMSO-d6) δ 10.43 (br. s., 1H), 8.76 (br. s., 1H), 7.21 - 7.29 (m, 2H), 7.07 - 7.16 (m, 2H), 7.04 (s, 1H), 2.87 (t, J = 7.63 Hz, 2H), 2.61 - 2.69 (m, 1H), 2.53-2.47 (m, 1H), 2.33 (t, J = 7.77 Hz, 2H), 1.61 - 1.67 (m, 1H), 1.56 (ddd, J = 4.69, 6.23, 8.73 Hz, 1H); 13C NMR (125 MHz, DMSO-d6) δ 170.1, 168.1, 161.7, 159.8, 155.0, 137.0, 136.9, 127.6, 127.5, 115.1, 115.0, 111.8, 31.6, 27.9, 26.9, 25.3, 19.7

Compound 37: N-Hydroxy-3-(2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide

White solid in 39% yield. 1H NMR (500 MHz, DMSO-d6) δ 10.39 (br. s., 1H), 8.80 (br. s., 1H), 7.44 - 7.47 (m, 2H), 7.38 - 7.41 (m, 2H), 7.33 - 7.35 (m, 1H), 6.97 (s, 1H), 2.84 (t, J = 7.63 Hz, 2H), 2.26 - 2.34 (m, 2H), 1.59 - 1.62 (m, 2H), 1.36 - 1.39 (m , 2H); 13C NMR (125 MHz, DMSO-d6) δ 175.3, 168.1, 155.0, 141.9, 130.2, 128.6, 125.5, 112.9, 31.6, 29.4, 26.9, 18.3, 17.9.

Compound 38: N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide

White solid in 12% yield. 1H NMR (400 MHz, DMSO-d6) δ 8.07-8.09 (d, J = 7.2 Hz, 2H), 7.60-7.63 (t, J = 7.2 Hz, 1H), 7.51-7.55 (t, J = 7.6 Hz, 2H), 6.84 (s, 1H), 2.85-2.89 (t, J = 7.6 Hz, 2H), 2.33-2.37 (t, J = 7.2 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.7, 165.4, 158.7, 150.6, 132.9, 132.6, 129.0, 128.5, 108.2, 32.0, 27.4.

Compound 39: 4-Fluoro-N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide

White solid in 19% yield. 1H NMR (400 MHz, DMSO-d6) δ 12.64 (br. s., 1H), 10.49 (br. s., 1H), 8.81 (br. s., 1H), 8.17 (dd, J = 5.56, 8.38 Hz, 2H), 7.35 (t, J = 8.74 Hz, 2H), 6.84 (s, 1H), 2.88 (t, J = 7.58 Hz, 2H), 2.36 (t, J = 7.70 Hz, 2H); 13C NMR (100 MHz, DMSO-d6) δ 168.4, 165.9, 164.1, 163.4, 158.4, 150.1, 131.1, 131.0, 128.8, 115.8, 115.6, 107.9, 31.7, 27.0.

Experimental Example 1. Confirmation of HDAC6 inhibitory activity of compounds 20 to 39

In order to confirm the inhibitory activity of compounds 20 to 39 according to Example 3 on HDAC1 and HDAC6, BPS bioscience assay kit (Fluorogenic HDAC1 assay kit, Fluorogenic HDAC6 assay, BPS number 50061, 50076) was used. Assay followed the manufacturer's protocol. The final reactant volume of 100㎕ was divided into 50㎕ each and distributed to a 96 well plate and measured using Glomax multi plus (Promega). All enzyme reactions were carried out in a 37℃ water bath for 30 minutes, and DMSO in the final reactant was carried out according to the following procedure. The concentration was diluted to 0.1%. The inhibitory activity test was repeated at least three times, and IC ₅₀ was calculated using GraphPad Prism 5 (Table 4).

Name R X n HDAC1 IC ₅₀ (nM) HDAC6 IC ₅₀ (nM) Selectivity Index
(HDAC1 IC ₅₀ /HDAC6 IC ₅₀ ) 20 - 2 2235±549.0 31.45±19.48 71.07 21 H 0 N.D. 44974±24500 - 22 H 2 9013±3153 63.84±24.17 141.18 23 H 3 4879±717.6 1121±534.5 4.35 24 F 2 4283±623.0 57.15±28.49 75.12 25 Br 2 21355±9352 247.8±106.8 86.18 26 OMe 2 12219±6526 25.56±11.47 478.05 27 - 2 6819±1475 405.3±164.0 16.82 28 - 2 3801±1267 86.42±19.04 43.98 29 H 2 14762±6422 91.2±28.97 161.9 30 F 2 10190±2677 69.92±19.21 145.7 31 H 2 2427±1399 36.65±21.41 66.22 32 F 2 10982±2175 211.5±82.84 51.92 33 - 2 N.D. 15705±7386 - 34 - 2 5168±387.1 165.2±28.32 31.28 35 H 2 1984±730.6 55.24±18.50 35.92 36 F 2 1480±159.1 79.41±22.29 18.64 37 2 1341±208.6 67.01±13.80 20.01 38 H 2 11408±1599 592.8±112.8 19.24 39 F 2 11467±3162 423.3±223.0 27.09

As a result, it was confirmed that compounds 22 and 26 had excellent selectivity for HDAC6 in that the IC ₅₀ value for HDAC6 was significantly lower than that for HDAC1. In particular, compound 26 showed the lowest IC ₅₀ value for HDAC6 (25.56 ± 11.47) among the tested compounds, but the highest IC ₅₀ value for HDAC1 (12219 ± 6526), making it the most effective as a selective inhibitor of HDAC6. It was found that

Experimental Example 2. Evaluation of inhibitory activity of compounds 22 and 26 by HDAC subtype

According to the results of Experimental Example 1, the inhibitory activity for each HDAC subtype of compounds 22 and 26, which were confirmed to have the best selective inhibitory activity, was confirmed. Specifically, Reaction Biology was commissioned to investigate the inhibitory activity of compounds 22 and 26 against HDAC1 to 11 (Table 5), and the control group was Pan-HDAC inhibitor, which is known to inhibit class I and II HDAC subtypes. Trichostatin A (TSA ^a ) was used. Compounds were tested in singlet, IO-dose and three-fold serial dilutions, blank spaces indicate inhibition or compound activity that did not fit the IC ₅₀ curve.

target Compound IC ₅₀ (M) TSA ^a 9a Compound 32 Compound 22 Compound 26 HDAC1 7.65E-09 >1.00E-05 4.65E-06 8.87E-06 >1.00E-5 HDAC2 2.82E-08 >1.00E-05 6.42E-06 - - HDAC3 1.50E-08 - 4.50E-06 7.70E-06 >1.00E-5 HDAC4 Not tested - 8.86E-05 - - HDAC5 Not tested - 3.34E-05 - - HDAC6 2.63E-09 5.67E-07 3.67E-08 5.34E-08 4.34E-08 HDAC7 Not tested - 2.04E-05 - - HDAC8 5.50E-07 1.54E-06 3.64E-06 4.46E-06 5.79E-06 HDAC9 Not tested - 1.26E-05 - - HDAC10 Not tested - 3.45E-06 Not tested Not tested HDAC11 3.35E-06 - 7.15E-06 2.48E-06 1.08E-06 * - means no activity

As a result, compounds 22 and 26 were confirmed to exhibit selective activity against HDAC subtypes. In particular, compound 26 showed selective inhibitory activity on HDAC1, 3, 6, 8, and 11. The control group, TSA ^a , is an inhibitor known to chelate zinc ions at the active site of HDAC, thus preventing histone unpacking, thereby strongly inhibiting HDAC, and to have multiple antitumor effects. Compound 26 is TSA and IC ₅₀ The values were confirmed to be close, and the inhibitory activity against HDAC6 was found to be similar, so it was confirmed to have an excellent HDAC inhibitory effect.

In addition, the selectivity for HDAC6 of compounds 22 and 26 compared to the base materials HDAC inhibitor 9a and compound 32 was confirmed, as shown in Table 6 below.

division HDAC1 IC ₅₀ (M) HDAC6 IC ₅₀ (M) Selectivity Index
(HDAC1 IC ₅₀ /HDAC6 IC ₅₀ ) 9a >1.00E-05 5.67E-07 127 Compound 32 4.65E-06 3.67E-08 117 Compound 22 8.87E-06 5.34E-08 168 Compound 26 >1.00E-05 4.34E-08 230

As a result, it was found that Compound 22 and Compound 26 had superior selectivity for HDAC6 compared to Compound 9a and Compound 32. In particular, in the case of compound 26, the selectivity for HDAC6 compared to 9a and compound 32 was found to be at least 1.8 to 1.96 times, and the activity of compound 26 as a selective inhibitor of HDAC6 was confirmed to be about twice as effective as that of 9a and compound 32. It has been done.

Experimental Example 3. Confirmation of in silico docking model and molecular dynamics simulation of compound 26

Experimental Example 3-1. Confirmation of in silico docking model of compound 26

In order to analyze the structure-activity correlation of Compound 26, which was confirmed to have the best selective inhibitory effect on HDAC6 in Experimental Examples 1 and 2, an in silico docking model with the HDAC6 X-ray crystal structure was confirmed as a result of the HDAC enzymatic assay. Specifically, the HDAC6 crystal structure was confirmed using PDB code 5EDU, the key residues of HDAC6 and the overall structure are shown in white, the crystal structure ligand trichostatin A is shown in yellow, and compound 26 is shown in pink, and docking was performed using Schrodinger XP-Glide. Model analysis was performed (Figure 1).

As a result, the cap group of Compound 26, that is, the thiazole group of Compound 26, consists of a structure capped with an ester group or acetamide group on one side and a naphthalene cap group on the other side, and the existing ring linker It was confirmed that HDAC6 selectivity further increased when the connection between the double bond C chain of the trans structure was extended to the naphthalene cap, and in silico for HDAC1 (PDB code: 4BKX) and HDAC6 (PDB code: 5EDU) This was reconfirmed by performing docking analysis. In the docking model, it was shown that the trans double bond structure anchors the cap structure of the inhibitor to a position that shows residue differences between subtypes (L3 pocket, Tyr204 in HDAC1, Phe679 in HDAC6). For this reason, the double bond structure is important for HDAC6 selectivity. It was confirmed to play a role.

Experimental Example 3-2. Molecular dynamics simulation analysis of compound 26

Molecular dynamics simulation analysis was performed based on the docking model for compound 26-HDAC6 obtained in Experimental Example 3-1. Specifically, a 500ns simulation was performed using GROMACS, and the interacting key residues and energy contribution were calculated using PyContact and g_mmpbsa software in the 50-500ns period (Figure 2).

As a result, the interaction between the naphthalene cap group and the HDAC6 L3 pocket was reconfirmed. In addition, it was confirmed that the cap group of compound 26 was strongly bound to the L3 pocket during a 500 ns simulation, and through comparison with a derivative with a flexible cap group, it was confirmed that the rigid cap group plays an important role in determining HDAC6 selectivity. In addition, it was confirmed that the methoxy substituent of Compound 26 is capable of additional binding through electrostatic interaction with Arg673, but since there is no residue in HDAC1 that can bind through the above interaction, but it exists in HDAC6, Compound 26's ability to bind to HDAC6 It was confirmed that selectivity was excellent.

Experimental Example 4. Confirmation of inhibitory activity of compound 26 against HDAC

To confirm the inhibitory activity of compound 26 against HDAC, a western blot experiment was performed. Specifically, SH-SY5Y cells were distributed in a 6 well plate and treated with compound 26 at concentrations of 0.03, 0.1, 0.3, 1, and 3 μM. After 24 hours, proteins were obtained from cells and western blot was performed using SDS-PAGE. In order to confirm changes in the activity of HDAC1, which mainly exists inside the nucleus, changes in acetylation of histone proteins were confirmed using Anti-histone H3 antibody (Abcam ab1791) and Anti-acetylated histone H3 antibody (Milipore 06-599). In addition, changes in the activity of HDAC6, which mainly exists in the cytoplasm, were confirmed by acetylation of tubulin protein using Anti-α tubulin (Abcam ab52866) and Anti-acetylated α tubulin (Abcam ab179484) (Figure 3).

As a result, Compound 26 was found to have a higher inhibitory activity against HDAC6 than HDAC1, showing excellent effectiveness as a selective inhibitor of HDAC6.

Experimental Example 5. Confirmation of superior inhibitory activity of Compound 26 on HDAC6 compared to SAHA

To confirm the superior inhibitory activity of compound 26 on HDAC6 compared to SAHA, a western blot experiment was performed. Specifically, HeLa cells were distributed in a 6 well plate and treated with compound 26 at concentrations of 0.3, 1, and 3 μM, and SAHA at concentrations of 0.3 and 3 μM was used as a control. After 24 hours, proteins were obtained from cells and western blot was performed using SDS-PAGE. Changes in the activity of HDAC6, which mainly exists in the cytoplasm, were confirmed by acetylation of tubulin protein using Anti-α tubulin (Abcam ab52866) and Anti-acetylated α tubulin (Abcam ab179484).

As a result, SAHA, a pan-HDAC subtype inhibitor, increased the acetylation of both tubulin and histone proteins, while compound 26 increased only the acetylation of tubulin proteins, showing that compound 26 was selective for HDAC6 compared to SAHA. It was confirmed that the inhibitory activity was excellent (Figure 4).

Overall, the derivative according to the present invention was found to have inhibitory activity against HDAC subtypes, and in particular, superior inhibitory activity against HDAC6 compared to other subtypes. This selective inhibitory activity is achieved by binding the cap group of compound 26 to a specific part of HDAC6. Since it was confirmed that this is because it forms a strong bond with , the correlation between the structure and activity of the compound of the present invention was identified. According to these results, the compound according to the present invention can be presented as a novel HDAC6 selective inhibitor with a new core structure, and is expected to be useful in preventing or treating HDAC6-mediated diseases.

The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims (6)

A compound represented by the following formula (1), an isomer thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

In the above formula, R is,
, , , , , , , , , , and Any one selected from the group consisting of,
X ₁ is any one selected from the group consisting of H, F, Br and OMe,
The X ₂ is H or F,
The n is an integer of 0, 2, or 3.
The method of claim 1, wherein the compound is:
N-hydroxy-3-(2-(naphthalen-1-yl)thiazol-4-yl)propanamide (20),
N-hydroxy-2-(naphthalen-2-yl)thiazole-4-carboxamide (21),
N-hydroxy-3-(2-(naphthalen-2-yl)thiazol-4-yl)propanamide (22),
N-hydroxy-4-(2-(naphthalen-2-yl)thiazol-4-yl)butanamide (23),
N-hydroxy-3-(2-(6-fluoronaphthalen-2-yl)thiazol-4-yl)-propanamide (24),
N-hydroxy-3-(2-(6-bromonaphthalen-2-yl)thiazol-4-yl)-propanamide (25),
N-hydroxy-3-(2-(6-methoxynaphthalen-2-yl)thiazol-4-yl)propanamide (26),
N-hydroxy-3-(2-(quinolin-2-yl)thiazol-4-yl)propanamide (27),
N-hydroxy-3-(2-(quinolin-3-yl)thiazol-4-yl)propanamide (28),
N-hydroxy-3-(2-([1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide (29),
N-hydroxy-3-(2-(4'-fluoro-[1,1'-biphenyl]-4-yl)thiazol-4-yl)-propanamide (30),
N-hydroxy-3-(2-(4-phenoxyphenyl)thiazol-4-yl)propanamide (31),
3-(2-(4-(4-fluorophenoxy)phenyl)thiazol-4-yl)-N-hydroxypropanamide (32),
(E)-N-hydroxy-3-(2-(1-phenylprop-1-en-2-yl)thiazol-4-yl)propanamide (33),
(E)-N-hydroxy-3-(2-(2-phenylprop-1-en-1-yl)thiazol-4-yl)propanamide (34),
N-hydroxy-3-(2-(2-phenylcyclopropyl)thiazol-4-yl)propanamide (35),
3-(2-(2-(4-fluorophenyl)cyclopropyl)thiazol-4-yl)-N-hydroxypropanamide (36),
N-hydroxy-3-(2-(1-phenylcyclopropyl)thiazol-4-yl)propanamide (37),
N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (38); and
4-Fluoro-N-(4-(3-(hydroxyamino)-3-oxopropyl)thiazol-2-yl)benzamide (39)
A compound represented by Formula 1, which is any one compound selected from the group consisting of, an isomer thereof, or a pharmaceutically acceptable salt thereof.
A pharmaceutical composition for preventing or treating HDAC6-mediated diseases, comprising the compound of claim 1, an isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.
According to paragraph 3,
The composition is a pharmaceutical composition for preventing or treating HDAC6-mediated diseases, characterized in that it selectively inhibits HDAC6.
According to paragraph 3,
The HDAC6-mediated diseases include organ transplant rejection, diseases related to abnormal function of lymphocytes, cell proliferative diseases, inflammatory diseases, autosomal dominant diseases, genetically related metabolic diseases, autoimmune diseases, cancer diseases, neurological diseases, hypertrophy, heart failure, A pharmaceutical composition for preventing or treating HDAC6-mediated diseases, which is at least one selected from the group consisting of eye diseases and neurodegenerative diseases.
A method for preventing or treating HDAC6-mediated diseases, comprising administering the composition of claim 3.