KR100562081B1 - A pharmaceutical composition containing Novel 2-oxo-pyrrol derivative compound I for treating cancer disease - Google Patents

Abstract

The present invention provides a pharmaceutical composition for the treatment of cancer diseases comprising the novel 2-oxo-pyrrole derivative (I) compound having excellent anticancer activity.

Since the compound according to the present invention shows a strong inhibitory activity against histone deacetylase (HDAC), and also shows a strong growth inhibitory action against cancer cell lines, the composition is a drug for the prevention and treatment of cancer diseases Available.

Anticancer, cancer disease, cell growth inhibition, histone deacetylase

Description

 A pharmaceutical composition containing Novel 2-oxo-pyrrol derivative compound (I) for treating cancer disease} including a novel 2-oxo-pyrrole derivative (I) compound having anticancer activity

The present invention relates to a composition for the treatment of cancer diseases comprising a novel 2-oxopyrrole derivative (I) compound having anticancer activity.

Cancer is characterized by "uncontrolled cell growth," which results in the formation of cell masses called tumors that infiltrate surrounding tissues and, in severe cases, metastasize to other organs of the body. Academia is also called neoplasia. Cancer is an intractable chronic disease that, even if treated with surgery, radiation, and chemotherapy, in many cases does not heal fundamentally, suffers the patient, and ultimately leads to death. More than 20 million people worldwide suffer from cancer, and more than 6 million people die of cancer every year, and 11 million people are expected to die by 2020, so cancer is an important disease that needs to be addressed as soon as possible. to be. Cancer varies from country to country, but in developed countries and Korea accounts for more than 20% of all deaths. However, despite many efforts, the exact cause or mechanism of cancer development is still unknown. There are many factors that cause cancer, but they can be divided into internal and external factors. It is not known exactly how normal cells are transformed into cancer cells, but at least 80-90% is known to be influenced by external factors such as environmental factors. Internal factors include genetic factors, immunological factors, and external factors include chemicals, radiation, and viruses. Genes involved in the development of cancer include oncogenes and tumor suppressor genes, which occur when the balance between them is broken down by internal or external factors described above.

Histone is a basic protein that binds to DNA in the nucleus of nucleated cells. Reversible acetylation occurs in the ε-amino group of lysine residues at specific positions in each molecule of histone. This histone acetylation reaction is associated with chromatin (chromatin) formation of higher structures, cell division cycles, and the like, and is known to be involved in the regulation of gene information expression with nonhistone proteins.

In vivo, the acetylation of histones is balanced by histone acetyltransferase and histone deacetylases (HDAC). have. To date, several histone deacetylase inhibitors have been published. Changes in histone acetylation levels can be modulated by compounds that inhibit histone deacetylase activity, which compounds known to date depend on their structure; 1) short chain fatty acids; Butyrate [Newmark et al., Cancer Lett . 78 , pp1-5, 1994], 2) hydroxamic acids structure; Trichostatin A, suberoylanilide hydroxamic acid (SAHA) and oxamflatin [Tsuji et al., J. Antibiot. (Tokyo) 29 , pp 1-6, 1976; Richon et al., Proc. Natl. Acad. Sci. USA 95 , pp 3003-3007, 1998; Kim et al., Oncogene 18 , pp2461-2470, 1999], 3) 2-amino-8-oxo-9,10-epoxydecanoyl (2-amino-8-oxo-9, 10-epoxy-decanoyl, AOE Cyclic tetrapeptide structure containing; Trapoxin A [Kijima et al., J. Biol. Chem. 268 , 22429-22435, 1993], 4) cyclic tetrapeptide structures without AOE; FR901228 and apicidin [Nakajima et al., Exp. Cell Res . 241 , pp 126-33,1998; Darkin-Rattray et al., Proc. Natl. Acad. Sci. USA 93 , pp13143-13147, 1996], 5) Benzamides structure; MS-27-275 by Saito et al., Proc. Natl. Acad. Sci. USA 96 , pp 4592-4597., 1999].

These compounds are known to inhibit human histone deacetylases to induce hyperacetylation of histone proteins, thereby overexpressing certain protein populations such as tumor suppressors and thereby inhibiting the growth of cancerous cells or killing cancer cells. In addition, it has been known that animal models suppress tumor growth. To date, the only example of such compound that has been used with clinical approval is butyrate. However, butyrate is not an ideal compound such as requiring a high concentration (millimolar unit, mM) to inhibit histone deacetylase and activating other enzymes in addition to histone deacetylase. Therefore, clinical studies of compounds with other structures capable of selectively inhibiting only histone deacetylases at low concentrations (micromolar units, μM) are actively underway, among which FR901228 is a phase 1 clinical trial. Cancer Institute). According to this fact, compounds capable of selectively inhibiting histone deacetylase are highly likely to be developed as breakthrough drugs that can induce growth inhibition and death of cancer cells.

Cancer is classified into blood cancer and solid cancer, and it occurs in almost every part of the body such as lung cancer, stomach cancer, breast cancer, oral cancer, liver cancer, uterine cancer, esophageal cancer, and skin cancer. Among the methods used to treat these malignancies, chemotherapy agents, except surgery or radiation therapy, are collectively called anticancer agents, and most of them show anticancer activity by inhibiting the synthesis of hexane. Chemotherapeutic agents are largely classified into antimetabolites, alkylating agents, antimitotic drugs, and hormones, and folates that inhibit the metabolic processes necessary for the proliferation of cancer cells. Derivatives (methotrexate), purine derivatives (6-mercaptopurine, 6-thioguanine), pyrimidine derivatives (5-fluorouracil, Cytarabine), etc.Introducing alkyl groups to guanine of DNA to modify the structure of DNA Alkylating agents that exert the effect include nitrogen mustard compounds (chlorambucil, cyclophosphamide), ethyleneimine compounds (thiotepa), alkylsulfonate compounds (busulfan), nitrosourea compounds (carmustine), triazene compounds (dacarbazine) There is. Mitosis inhibitors that inhibit mitosis by blocking mitosis as anti-mitotic drugs include plant drugs such as actinomycin D, doxorubicin, bleomycin, and anticancer drugs such as mitomycin, vincristine, and vinblastine. Alkaloids, taxoids including mitosis inhibitors including taxane rings, and the like. In addition, hormones such as corticosteroids and progesterone and platinum-containing compounds such as cisplatin are used as anticancer agents.

The biggest problem with chemotherapeutic agents is drug resistance, which, despite the initial successful response by anticancer agents, is a major factor that eventually causes treatment to fail. In addition to researching the cause of drug resistance, the development of anti-cancer drugs with new mechanisms is needed to treat cancers resistant to existing drugs. Current anticancer drugs include drug resistance blockers, angiogenesis inhibitors, tumor metastasis inhibitors, and drugs targeting gene expression. Among these, a study for developing a compound that inhibits the activity of histone deacetylase enzyme as an anticancer agent is currently in progress. Histone deacetylase is an enzyme involved in gene expression by cells. When histone deacetylase of cancer cells is inhibited, phenomena such as differentiation, cell death and cell cycle blocking occur.

The inventors of the present invention conducted a study to develop a substance showing a strong inhibitory activity against the growth of tumor cells using histone deacetylase as a functioning point, and the 2-oxo-pyrrole derivative compound (I) of the present invention was excellent in histone diacetylase. The present invention was found to exhibit acetylase inhibitory activity and growth inhibitory activity of tumor cells.

An object of the present invention is to provide a composition for the treatment of cancer diseases comprising a novel 2-oxo pyrrole derivative (I) compound exhibiting excellent anticancer effects.

In order to achieve the above object, the present invention contains a 2-oxo pyrrole derivative (I) compound having a structure of the general formula (I) or a pharmacologically acceptable salt thereof as an active ingredient, which is useful for the treatment of cancer diseases. Pharmaceutical compositions are provided:

(Ⅰ)

(Ⅰ)

Where

,

이고,X is -OH, -NHOH, -NHOCH ₂ Ph,

,

ego,

n is an integer from 1 to 5,

R is an optionally substituted substituent, optionally a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms substituted with R`, an alkenyl group, an alkynyl group or an allyl group,

R` is a substituent selected from a heterocyclic group or an aromatic aryl group, preferably a thiophenyl group, a naphthyl group, a pyrrole group, a furyl group and a biphenyl group,

)은 단일 결합 또는 이중 결합을 의미한다.dotted line(

) Means a single bond or a double bond.

General formula Particularly preferred groups of compounds of (I) include the following compounds and pharmaceutically acceptable salts thereof:

N-hydroxy-3- (1-naphthalen-2-yl-methyll-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (1e '),

N-hydroxy-3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (2h '),

3- (1-allyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -N-hydroxy-propionamide (3h '),

N-hydroxy-3- [1- (2-naphthalen-1-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (4n '),

N-hydroxy-3- [1- (2-naphthalen-2-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (5n '),

N-hydroxy-3- [2-oxo-1- (2-thiophen-2-yl-ethyl) -2,5-dihydro-1H-pyrrol-3-yl] -propionamide (6n '),

3- [1- (3-biphenyl-4-yl-propyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -N-hydroxy-propionamide (7w '),

N-hydroxy-3- [1- (3-naphthalen-2-yl-propyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (8w ') .

The present invention also provides a method for preparing the compound of Formula (I) and a pharmaceutical composition comprising the same as an active ingredient.

The compounds of the present invention represented by the general formula (I) may be prepared with pharmaceutically acceptable salts and solvates according to conventional methods in the art.

As salts are acid addition salts formed with pharmaceutically acceptable free acids. Acid addition salts are prepared by conventional methods, for example by dissolving a compound in an excess of aqueous acid solution and precipitating the salt using a water miscible organic solvent, such as methanol, ethanol, acetone or acetonitrile. Equal molar amounts of the compound and acid or alcohol (eg, glycol monomethylether) in water can be heated and the mixture can then be evaporated to dryness or the precipitated salts can be suction filtered.

In this case, organic acids and inorganic acids may be used as the free acid, hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid, etc. may be used as the inorganic acid, and methanesulfonic acid, p -toluenesulfonic acid, acetic acid, trifluoroacetic acid, etc. may be used as the organic acid. , Citric acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, manderic acid, propionic acid, citric acid, lactic acid, glycolic acid, gluconic acid (gluconic acid), galacturonic acid, glutamic acid, glutaric acid (glutaric acid), glucuronic acid (glucuronic acid), aspartic acid, ascorbic acid, carbonic acid, vanic acid, hydroiodic acid may be used.

Bases can also be used to make pharmaceutically acceptable metal salts. An alkali metal or alkaline earth metal salt is obtained by, for example, dissolving a compound in an excess of alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and then evaporating and drying the filtrate. At this time, as the metal salt, it is particularly suitable to prepare sodium, potassium or calcium salts, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (for example, silver nitrate).

Pharmaceutically acceptable salts of the above general formula (I) include salts of acidic or basic groups which may be present in compounds of general formula (I), unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulphate, phosphate, hydrogen phosphate, dihydrogen Phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p -toluenesulfonate (tosylate) salts. It can be prepared through the process.

Another object of the present invention is to provide a method for preparing the compound of Formula (I), which may be chemically synthesized by the method shown in the following schemes, but is not limited thereto.

The following reaction schemes represent the preparation steps of the representative compounds of the present invention. The various compounds of the present invention may be prepared by small changes such as changing the reagents, solvents, and reaction sequences used in the synthesis of Schemes 1 to 4. have. Some compounds of the present invention have been synthesized according to procedures not included in the scope of Schemes 1-4, and detailed synthesis procedures for these compounds are described in their respective examples.

Scheme (1) shows a four-step preparation process for preparing compound (e) from the commercially available amine compound (a) as a starting material.

In the first step, compound (b) is prepared by reacting compound (a) with allyl bromide in an organic solvent in the presence of a Hunig base. At this time, acetonitrile (MeCN), dichloromethane, and the like may be used as the organic solvent, and diethylisopropylamine, which is Hunig's base, may be used. Hunig base may be used in 2 to 3 equivalents relative to the starting compound (a), and their reaction may be carried out at temperatures ranging from room temperature to 0 ° C.

In the second step, compound (c) is reacted with monoacid in an organic solvent in the presence of compound (b) obtained in step 1 in the presence of 1- [3- (dimethylamino) propyl] -3-ethylcarbodiimide (EDC). Manufacture. In this case, methylene chloride, tetrahydrofuran, or the like may be used as the organic solvent. In this case, the mono acid is preferably 2-methylene-pentanedioic acid-5-methyl ester, and the amount thereof may be used in an amount of 1 to 1.2 equivalents based on compound (b), and their reaction may range from room temperature to 0 ° C. It can be carried out at a temperature of.

In a third step, compound (c) is then converted to compound (d) in an organic solvent in the presence of a groove (I) catalyst (Grubb's (I) catalysis) such as a ruthenium catalyst. In this case, the amount of the catalyst is preferably used in 0.02 to 0.1 equivalents based on the compound (c), these reactions can be carried out at a temperature in the range of room temperature to 0 ℃.

Subsequently, in the fourth step, the compound (d) may be reacted with an amine salt in an alcohol solvent to obtain a compound (e) in which the substituent X in the compound of the formula according to the present invention is -NHOH. The amine salt is preferably potassium hydroxyamide, the amount of which is preferably used in an amount of 2 to 3 equivalents based on compound (d), and the reaction thereof may be performed at a temperature ranging from room temperature to 0 ° C.

Scheme (2) shows a process for obtaining compound (n) from compound (d) in scheme (1).

Compound (d) in Scheme (1) is reacted with triethylsilane in trifluoroacetic acid to obtain compound (1), wherein triethylsilane is preferably used in a ratio of 1 to 1.5 equivalents relative to compound (d). . In addition, compound (l) in Scheme (5) is added with hexamethyldisilylazide sodium (NaHMDS) in a tetrahydrofuran solvent, and then allyl bromide or methyl iodine is added to obtain compound (m). In a second step, compound (m) is added to an amine salt in methanol solution to obtain compound (n). The amount of NaHMDS used in the reaction is preferably 1 to 1.5 equivalents based on compound (1), and 1 to 1.5 equivalents on allyl bromide.

Scheme (3) shows a five-step process for preparing compound (y) using a commercially available alcohol compound (t) as a starting material.

In the first step, p -toluenesulfonylchloride, diisopropylethylamine and 4- (dimethylamino) pyridine were injected into the solution dissolved in methylene chloride by argon substitution of alkyl-substituted ethyl alcohol (t) at 0 ° C. After stirring for an hour, the mixture was stirred for 12 hours at room temperature.

In the second step, the compound (u) obtained in step 1 is dissolved in acetonitrile, and then allylamine and diisopropylethylamine are injected, followed by stirring at 80 ° C. for 6 hours.

In the third step, 2-methylene-pentanedioic acid-5-methyl ester and 1- (3- (dimethylamino) propyl)-are dissolved in the methylene chloride solution dissolved by argon substitution of the compound (v) obtained in the second step. 3-ethylcarbodiimide and 4- (dimethylamino) pyridine were injected and stirred at room temperature for 10 hours.

In the fourth step, the compound (w) is converted into the compound (w) in the organic solvent in the presence of the groove (I) catalyst (Grubb's (I) catalysis) such as the compound (v) and the ruthenium catalyst obtained in the third step.

In the fifth step, the compound (x) obtained in the fourth step may be reacted with an amine salt in an alcohol solvent to obtain a compound (y) in which the substituent X in the compound of the formula of the present invention is -NHOH. The amine salt is preferably potassium hydroxyamide, the amount of which is preferably used in an amount of 2 to 3 equivalents based on compound (x), and the reaction thereof may be performed at a temperature ranging from room temperature to 0 ° C.

Scheme (4) shows an eight-step process for preparing compound (ah) using aldehyde (z), which is readily available commercially.

In the first step, compound (a) is prepared by reacting compound (z) with Wittig reagent in an organic solvent. In this case, dichloromethane (CH ₂ Cl ₂ ) may be used as the organic solvent. The Witig reagent may be used in 1.5 to 2 equivalents based on the starting compound (z), and the reaction thereof may be performed at a temperature in the range of 60 to 70 ° C.

In the second step, the compound (aa) obtained in step 1 is treated with Pd-C in an alcohol solvent to obtain compound (ab), wherein Pd-C is used in an amount of 0.1 to 0.2 equivalents based on compound (aa). desirable.

In the third step, compound (ab) is reacted with lithium aluminum hydride (LAH) to obtain compound (ac). The organic solvent may use THF and may be carried out at a temperature in the range of 0 ℃.

In the fourth step, the compound (ac) obtained in step 3 is argon-substituted and reacted with p -toluenesulfonylchloride, diisopropylethylamine and 4- (dimethylamino) pyridine in a solution dissolved in dichloromethane to give compound (ad). At this time, the reaction starts at 0 ℃ and after several hours can be reacted at room temperature for more than 12 hours.

In the fifth step, the compound (ad) obtained in step 4 was dissolved in acetonitrile (MeCN), and then allylamine and Hunig's diisopropylethylamine were injected, followed by stirring at 80 ° C. for 6 hours. Get

In the sixth step, 2-methylene-pentanedioic acid-5-methyl ester, 1- (3- (dimethylamino) propyl)-in a dichloromethane solution dissolved by argon substitution of the compound (ae) obtained in the fifth step. Injecting 3-ethylcarbodiimide and 4- (dimethylamino) pyridine and stirring at room temperature for 10 hours to obtain a reactant (af).

In the seventh step, the compound (af) obtained in the sixth step is converted to the compound (ag) in an organic solvent in the presence of a groove (I) catalyst (Grubb's (I) catalysis) such as a ruthenium catalyst.

In the eighth step, the compound (ag) obtained in the seventh step may be reacted with an amine salt in an alcohol solvent to obtain a compound (ah) in which the substituent X in the compound of the formula of the present invention is -NHOH. The amine salt is preferably potassium hydroxyamide, the amount of which is preferably used in an amount of 2 to 3 equivalents based on the compound (ag), and the reaction thereof may be performed at a temperature ranging from room temperature to 0 ° C.

Derivatives of the present invention from the above production method shows a strong inhibitory activity against histone deacetylase and also a strong growth inhibitory effect against cancer cell lines, showing an excellent anti-cancer effect, consequently to treat cancer-related diseases useful.

Accordingly, the present invention provides a pharmaceutical composition for treating cancer diseases comprising the compound of the general formula (I) as an active ingredient and a pharmaceutically acceptable carrier.

The cancer diseases include lung cancer, non-small cell lung cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, anal muscle cancer, colon cancer, breast cancer, and fallopian tube carcinoma , Endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penis cancer, prostate cancer , Chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brain stem glioma, pituitary adenoma The pharmaceutical composition of the present invention can be used for the treatment of cancer.

Compositions comprising a compound of the present invention may further comprise a suitable carrier, excipient or diluent according to conventional methods.

Carriers, excipients and diluents that may be included in the compositions of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The compositions comprising the compounds of the present invention are each formulated in the form of oral dosage forms, external preparations, suppositories, or sterile injectable solutions, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., in accordance with conventional methods. Can be used.

Specifically, when formulated, it may be prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrating agents, surfactants, and the like. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ), Lactose, gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc can also be used. Liquid preparations for oral use include suspensions, solvents, emulsions, and syrups.In addition to the commonly used simple diluents, water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. May be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

Preferred dosages of the compounds of the present invention depend on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, but may be appropriately selected by those skilled in the art. However, for the desired effect, the compound of the present invention may be administered in an amount of 0.0001 to 100 mg / kg, preferably in an amount of 0.001 to 100 mg / kg once to several times daily. The compound of the present invention in the composition should be present in an amount of 0.0001 to 10% by weight, preferably 0.001 to 1% by weight relative to the total weight of the total composition.

In addition, the pharmaceutical dosage forms of the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical composition of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Hereinafter, the present invention will be described in more detail based on the following Examples and Experimental Examples. However, the following Reference Examples and Examples are only for illustrating the present invention, but the scope of the present invention is not limited thereto.

Reference Example 1 Synthesis of 3- [1- (2,4-Dimethoxy-benzyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -N-hydroxypropionamide (1e )

Step 1. Synthesis of Allyl- (2,4-dimethoxybenzyl) amine (1b)

In a methylene chloride solution containing 2,4-dimethoxybenzylamine (500 mg, 3.33 mmol), allylbromide (0.320 mL, 3.66 mmol) and diisopropylethylamine (0.700 mL, 3.99 mmol) were stirred. After injection, the mixture was stirred at room temperature for 12 hours. The resulting mixture was neutralized with 10% sodium hydroxide, extracted with chloroform, washed with saturated brine solution, dried over magnesium sulfate and distilled under reduced pressure. The obtained primary compound was separated by column chromatography method (silica gel column, eluent methanol / chloroform = 1/9) to give the title compound in 40% yield (276 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.12 (d, J = 8.1 Hz, 1H), 6.44-6.39 (m, 2H), 5.99-5.86 (m, 1H), 5.21-5.09 (m, 2H ), 3.79 (d, J = 6.0 Hz, 6H), 3.74 (s, 2H), 3.23 (d, J = 6.0 Hz, 2H)

Step 2. Synthesis of 4- [allyl- (2,4-dimethoxy-benzyl) -carbamoyl] -pent-4-enoic acid methyl ester (1c)

Arylene was substituted with the compound obtained in the first step, 2-methylene-pentanedioic acid-5-methyl ester (253 mg, 1.6 mmol) and 1- (3- (dimethylamino) propyl in 0.5 M solution dissolved in argon. ) -3-ethylcarbodiimide (331 mg, 1.73 mmol) and 4- (dimethylamino) pyridine (48 mg, 0.39 mmol) were injected, followed by stirring at room temperature for 10 hours. The resulting mixture was washed with 5% hydrochloric acid solution (10 mL), extracted with ethyl acetate, washed with saturated brine solution, and washed again with saturated sodium carbonate solution (10 mL) and saturated brine. The organic layer was dried over magnesium sulfate, distilled under reduced pressure, and then the obtained primary compound was purified by column chromatography (silica gel column, eluent ethyl acetate / hexane = 1/2) to give the title compound in a yield of 70% (324 mg). Got it.

¹ H-NMR (300 MHz, CDCl ₃ ) δ 7.14 (s, 1H), 6.44 (d, 2H), 5.72 (s, 1H), 5.12 (s, 4H), 4.56-4.81 (m 2H), 3.91- 3.83 (m, 2H), 3.78 (d, J = 5.3 Hz, 6H), 3.65 (d, J = 1.4 Hz, 3H), 2.63 (t, J = 5.7 Hz, 2H), 2.54 (t, J = 5.4 Hz, 2H)

Step 3. Synthesis of 3- [1- (2,4-Dimethoxybenzyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionic acid methyl ester (1d)

Argon-substituted the compound (324 mg, 0.933 mmol) and the ruthenium catalyst (74 mg, 0.09 mmol) obtained in the second step were dissolved in methylene chloride solution (93 mL) and stirred at room temperature for 24 hours. The primary compound obtained by distillation of the solvent under reduced pressure was purified by column chromatography (silica gel column, eluent: ethyl acetate / hexane = 1/1) to obtain the title compound in 90% yield (268 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.11 (d, J = 9.0 Hz, 1H), 6.61 (br t, 1H), 6.43 (s, 1H), 6.40 (d, J = 2.7 Hz, 1H ), 4.56 (s, 2H), 3.78 (d, J = 5.4 Hz, 9H), 3.65 (s, 2H), 2.61 (s, 4H)

Step 4. Synthesis of 3- [1- (2,4-Dimethoxy-benzyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -N-hydroxypropionamide (1e)

NH ₂ OK (1.7 M suspension in methanol, 0.27 mL, 0.47 mmol) was added to a methanol solution of the compound (100 mg, 0.313 mmol) obtained in the third step at 0 ° C., and the mixture was stirred at room temperature for 4 hours. The mixture was neutralized with acetic acid (0.020 mL), distilled under reduced pressure, and then filtered through solid with 10% methanol / chloroform and distilled under reduced pressure. The obtained primary compound was purified by column chromatography method (silica gel column, eluent: methanol / chloroform = 1/9) to obtain the title compound in 50% yield (50 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.04 (d, J = 8.1 Hz, 1H), 6.83 (s, 1H), 6.53-6.44 (m, 2H), 4.54 (s, 2H), 3.81 ( t, J = 2.0 Hz, 6H), 2.56 (t, J = 7.2 Hz, 2H), 2.34 (t, J = 7.4 Hz, 2H), 1.9 (s, 3H)

Reference Example 2. 3- {1- [2- (2-Fluoro-phenyl) -ethyl] -2-oxo-2,5-dihydro-1 H-pyrrol-3-yl} -N-hydroxy-propionamide Synthesis of (28y)

Step 1. Synthesis of toluene-4-sulfonic acid-2- (2-fluoro-phenyl) -ethyl ester (28u)

P -toluenesulfonylchloride (1.02 g, 5.35 mmol), diisopropylethylamine (1.24 mL) in a solution of 2- (2-fluoro-phenyl) -ethanol (500 mg, 3.57 mmol) in argon substitution in methylene chloride , 7.13 mmol), 4- (dimethylamino) pyridine (86mg, 0.71 mmol) were injected at 0 ° C., stirred for 6 hours, and then stirred at room temperature for 12 hours. The reaction product is neutralized with ammonium chloride, extracted with ethyl acetate and washed with saturated brine. After drying with magnesium sulfate and distilling under reduced pressure, the obtained primary compound was purified by column chromatography (silica gel column, eluent: methanol / chloroform = 1/7) to give the title compound in a yield of 70% (740 mg).

Step 2. Synthesis of Allyl- [2- (2-Fluoro-phenyl) -ethyl] -amine (28v)

After dissolving the compound (350 mg, 1.19 mmol) obtained in the first step in acetonitrile, allylamine (0.89 mL, 11.89 mmol) and diisopropylethylamine (0.31 ml, 1.78 mmol) were injected, and the mixture was dried at 80 ° C. Stir for time. The resulting mixture was neutralized with 10% sodium hydroxide, extracted with chloroform, washed with saturated brine solution, dried over magnesium sulfate and distilled under reduced pressure. The obtained primary compound was separated by column chromatography method (silica gel column, eluent 10% methanol / chloroform) to give the title compound in 66% yield (141 mg, 0.79 mmol).

Step 3. Synthesis of 4- {allyl- [2- (3-fluoro-phenyl) -ethyl] -carbamoyl} -pent-4-enoic acid methyl ester (28w)

2-methylene-pentanedioic acid-5-methylester (106 mg, 0.67 mmol), 1- (3- in the methylene chloride solution dissolved by argon substitution of the compound (100 mg, 0.56 mmol) obtained in the second step. (Dimethylamino) propyl) -3-ethylcarbodiimide (139 mg, 0.73 mmol) and 4- (dimethylamino) pyridine (20 mg, 0.17 mmol) were injected and stirred at room temperature for 10 hours. The resulting mixture was washed with 5% hydrochloric acid solution (10 mL), extracted with ethyl acetate, washed with saturated brine solution, and washed again with saturated sodium carbonate solution (10 mL) and saturated salt solution. The organic layer was dried over magnesium sulfate, and distilled under reduced pressure, and then the obtained primary compound was purified by column chromatography (silica gel column, eluent ethyl acetate / hexane = 1/2) to give the title compound in 72% yield (0.40 mmol, 128). mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.19-7.09 (m, 1H), 7.04-6.94 (m, 3H), 5.84 -5.57 (m, 1H), 5.13 (t, J = 10.7 Hz, 4H ), 5.06-4.94 (m, 2H), 3.79 (s, 2H), 3.62 (s, 4H), 3.53 (d, J = 5.4 Hz, 3H), 2.89 (d, J = 6.0 Hz, 3H).

Step 4. Synthesis of 3- {1- [2- (2-Fluoro-phenyl) -ethyl] -2-oxo-2,5-dihydro-1H-pyrrol-3-yl} -propionic acid methyl ester (28x)

Argon-substituted compounds (100 mg, 0.31 mmol) and ruthenium catalyst (27 mg, 0.03 mmol) obtained in the third step were dissolved in methylene chloride solution (31.3 mL) and stirred at room temperature for 24 hours. The primary compound obtained by distillation of the solvent under reduced pressure was purified by column chromatography method (silica gel column, eluent: ethyl acetate / hexane = 1/1) to give the title compound in a yield of 75% (69 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.16-7.12 (m, 2H), 7.03-6.93 (m, 2H), 6.59 (br t, 1H), 3.67 -3.65 (m, 4H), 3.62 ( s, 3H), 2.89 (t, J = 7.3 Hz, 2H), 2.56 (s, 4H).

Step 5. 3- {1- [2- (2-Fluoro-phenyl) -ethyl] -2-oxo-2,5-dihydro-1 H-pyrrol-3-yl} -N-hydroxy-propionamide ( 28y) synthesis

NH ₂ OK (1.7 M suspension in methanol, 0.38 mL, 0.65 mmol) was added to a methanol solution of the compound (38 mg, 0.13 mmol) obtained in the fourth step at 0 ° C., and the mixture was stirred at room temperature for 8 hours. The mixture was neutralized with acetic acid (0.02 mL), distilled under reduced pressure, and then filtered through solid with 10% methanol / chloroform and distilled under reduced pressure. The obtained primary compound was purified by column chromatography method (silica gel column, eluent: methanol / chloroform = 1/9) to give the title compound in 65% yield (25 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.19-7.08 (m, 2H), 7.02-6.92 (m, 2H), 6.69 (br t, 1H), 3.69 (s, 2H), 3.63 (t, J = 7.0 Hz, 2H), 2.87 (t, J = 7.0 Hz, 2H), 2.51 (t, J = 7.0 Hz, 2H), 2.25 (t, J = 7.3 Hz, 2H).

Reference Example 3. N-hydroxy-3- {1- [3- (4-methoxy-phenyl) -propyl] -2-oxo-2,5-dihydro-1 H-pyrrol-3-yl} -propion Synthesis of Amide (40ah)

Step 1. 3- p -Tolyl-acrylic acid methyl ester (40aa)

p -tolualdehyde (1 g, 8.30 mmol) and triphenylphosphanilidene-acetic acid methyl ester (4.16 g, 12.45 mmol) were dissolved in methylene chloride solution and stirred at 90 ° C. overnight. The resulting mixture was distilled under reduced pressure, and then ethyl acetate / hexane = 1/7 solution was added thereto, followed by stirring for 1 hour. If you have white salt, filter it out. The remaining solution was distilled under reduced pressure to give the title compound in 95% yield (1.39 g).

Step 2. 3- p -Tolyl-propionic acid methyl ester (40ab)

The flask containing the compound (1.39 g, 7.90 mmol) obtained in the first step was nitrogen-substituted and dissolved in methanol. Add a palladium catalyst, remove nitrogen and add hydrogen gas. After reaction at room temperature for about 1 hour and a half, the palladium catalyst was filtered off and purified by column chromatography method (silica gel column, eluent: ethyl acetate / hexane = 1/10) to give the title compound in 95% yield (1.24 g). Got it.

Step 3. 3- p -Tolyl-propan-1-ol (40ac)

After argon substitution of the compound (1.24 g, 6.95 mmol) obtained in the second step, 100 ml of tetrahydrofuran is injected. Lithium aluminum hydride was stirred at 27 ml 0 ° C. and then stirred for 2 hours. 3 ml of water, 3 ml of 1N NaOH, and 9 ml of water were injected into the reaction product, followed by stirring for about 30 minutes. Filter using a glass filter in a glass filter. The filtered solution was distilled under reduced pressure to obtain a compound that was purified by column chromatography (silica gel column, eluent: methanol / chloroform = 1/2) to obtain the title compound in 93% yield (6.46 mmol, 971 mg).

Step 4. Toluene-4-sulfonic acid-3- p -Tolyl-propyl ester (40ad)

Tosyl chloride (2.46 g, 13.0 mmol), diisopropylethylamine (3.40 ml, 19.40 mmol), 4- in a solution of methylene chloride obtained by argon substitution of the compound (971 mg, 6.46 mmol) obtained in the third step was 4- (Dimethylamino) pyridine (158 mg, 1.29 mmol) was injected at 0 ° C., stirred for 6 hours, and then stirred at room temperature for 12 hours. The reaction product is neutralized with ammonium chloride, extracted with ethyl acetate and washed with saturated brine. After drying over magnesium sulfate and distillation under reduced pressure, the obtained primary compound was purified by column chromatography (silica gel column, eluent: methanol / chloroform = 1/7) to obtain the title compound in 70% yield (1.30 g).

Step 5. Allyl- (3- p Synthesis of -tolyl-propyl) -amine (40ae)

The compound obtained in step 4 (1.30 g, 4.27 mmol) was dissolved in acetonitrile, and then allylamine (1.60 ml, 21.40 mmol) and diisopropylethylamine (0.97 ml, 5.50 mmol) were injected. Stir for time. The resulting mixture was neutralized with 10% sodium hydroxide, extracted with chloroform, washed with saturated brine solution, dried over magnesium sulfate and distilled under reduced pressure. The obtained primary compound was separated by column chromatography method (silica gel column, eluent 10% methanol / chloroform) to give the title compound in 85% yield (3.62 mmol, 687 mg,).

Step 6. 4- [allyl- (3- p -Tolyl-propyl) -carbamoyl] -pent-4-enoic acid methyl ester (40af)

Arylene-substituted compound (687 mg, 3.62 mmol) obtained in the fifth step was dissolved in methylene chloride 0.5 M solution, 2-methylene-pentanedioic acid-5-methylester (683 mg, 4.30 mmol), 1- ( 3- (dimethylamino) propyl) -3-ethylcarbodiimide (902 mg, 4.70 mmol) and 4- (dimethylamino) pyridine (133 mg, 1.09 mmol) were injected and stirred at room temperature for 10 hours. The resulting mixture was washed with 5% hydrochloric acid solution (10 mL), extracted with ethyl acetate, washed with saturated brine solution, and washed again with saturated sodium carbonate solution (10 mL) and saturated brine. The organic layer was dried over magnesium sulfate, distilled under reduced pressure, and then the obtained primary compound was purified by column chromatography (silica gel column, eluent ethyl acetate / hexane = 1/2) to give the title compound in 73% yield (2.60 mmol, 797). mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.05 (s, 4H), 5.70 (s, 1H), 5.16-5.07 (m, 4H), 3.94 (s, 2H), 3.64 (t, J = 3.3 Hz, 3H), 3.36 (s, 2H), 2.67-2.51 (m, 6H), 2.28 (s, 3H), 1.83 (t, J = 7.7 Hz, 2H)

Step 7. 3- [2-oxo-1- (3- p -Tolyl-propyl) -2,5-dihydro-1H-pyrrol-3-yl] -propionic acid methyl ester (40ag)

The compound (797 mg, 2.60 mmol) and the ruthenium catalyst (180 mg, 0.10 mmol) obtained in the sixth step were substituted with argon, dissolved in a methylene chloride solution (200 mL), and stirred at room temperature for 24 hours. The primary compound obtained by distillation of the solvent under reduced pressure was purified by column chromatography (silica gel column, eluent: ethyl acetate / hexane = 1/1) to obtain the title compound in a yield of 50% (391 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 6.96 (s, 4H), 6.56 (s, 1H), 3.67 (s, 2H), 3.55 (s, 3H), 3.37 (t, J = 7.2 Hz, 2H), 2.48 (t, J = 8.2 Hz, 6H), 2.19 (s, 3H), 1.76 (t, J = 7.6 Hz, 2H)

Step 8. N-hydroxy-3- [2-oxo-1- (3- p -Tolyl-propyl)-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (40ah)

NH ₂ OK (1.7 M suspension in methanol, 0.82 mL, 5.0 mmol) was added to a methanol solution of the compound (100 mg, 0.33 mmol) obtained in the seventh step at 0 ° C., followed by stirring at room temperature for 8 hours. The mixture was neutralized with acetic acid (0.020 mL), distilled under reduced pressure, and then filtered through solid with 10% methanol / chloroform and distilled under reduced pressure. The obtained primary compound was purified by column chromatography method (silica gel column, eluent: methanol / chloroform = 1/9) to give the title compound in 50% yield (50 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 7.21 (s, 4H), 6.95 (s, 1H), 3.96 (s, 2H), 3.60 (s, 2H), 2.72 (s, 5H), 2.45 ( s, 3H), 1.99 (s, 2H)

Example 1 Synthesis of N-hydroxy-3- (1-naphthalen-2-ylmethyll-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (1e ')

Compounds of Example 1 having physical properties as shown in Table 1 were prepared by performing a preparation process similar to the preparation method described in Reference Example 1.

Example Chemical structure NMR Spectrum or LC-MS Data One

RT: 3.93-5.93 (Mass: 311.2)

Example 2. Synthesis of N-hydroxy-3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (2h ')

Step 1. Synthesis of 3- (2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionic acid methyl ester (2f)

Of 3- [1- (2,4-dimethoxybenzyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionic acid methyl ester compound (1d) (200 mg, 0.63 mmol) Triethylsilane (0.100 mL, 0.63 mmol) was added to the trifluoroacetic acid (0.7 mL) solution, followed by heating at 0 ° C. for 1 hour. The solution was distilled under reduced pressure to remove the solvent, and then dissolved in chloroform (20 mL), and then the organic layer was washed with saturated sodium carbonate solution (5 mL) and saturated brine (5 mL). The organic layer was then dried over magnesium sulfate and then distilled under reduced pressure. The obtained primary compound was purified by column chromatography (silica gel column, eluent: ethyl acetate / methanol = 19/1) to give 3- (2-oxo-2,5-dihydro-1H-pyrrol-3-yl)-. Propionic acid methyl ester was obtained (yield 47%, 50 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 6.76 (br t, 1H), 3.89 (d, J = 1.3 Hz, 2H), 3.63 (t, J = 1.9 Hz, 3H), 2.58 (s, 4H ).

Step 2. Synthesis of 3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionic acid methyl ester (2 g)

NaHMDS (1.0 M in THF, 0.330 mL, 0.330 mM) was slowly injected into THF (0.6 mL) solution of the compound (3f) (50 mg, 0.295 mM) obtained in the first step at -79 ° C, and stirred for 30 minutes. It was. Dimethyl sulfate (0.03 mL, 0.359 mmol) was added to the reaction mixture, which was then stirred at 0 ° C. for 4 hours. Then, saturated ammonia chloride solution (2 mL) was added to the reaction mixture, and the mixture was diluted with ethyl acetate (7 mL). The organic layer was washed with ammonia chloride solution (2 mL) and saturated brine (2 mL), and then the organic layer was dried over magnesium sulfate and distilled under reduced pressure. The obtained primary compound was purified by column chromatography (silica gel column, eluent: ethyl acetate) to obtain 3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -methyl propionate. Ester compound (3 g) was obtained (yield 33%, 18 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 6.65 (br t, 1H), 3.81 (s, 1H), 3.64 (s, 3H), 3.01 (s, 3H), 2.60 (t, 4H).

Step 3. Synthesis of N-hydroxy-3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (2h)

NH ₂ OK (1.7 M suspension in methanol, 0.09 mL, 0.15 mmol) was added to a methanol solution of the compound (18 mg, 0.10 mmol) obtained in the second step at 0 ° C., and the mixture was stirred at room temperature for 1 hour. The mixture was neutralized with acetic acid (0.03 mL), distilled under reduced pressure, and then filtered through solid with 10% methanol / chloroform and distilled under reduced pressure. The obtained primary compound was purified by column chromatography method (silica gel column, eluent: ethyl acetate / methanol = 5/2) to give the title compound in 59% yield (11 mg).

¹ H-NMR (300 MHz, CDCl ₃ ), δ 6.76 (br t, 1H), 3.84 (s, 2H), 3.00 (s, 3H), 2.59 (t, J = 7.2 Hz, 2H), 2.42 (t , J = 7.2 Hz, 2H).

Compounds of Example 3 having physical properties as shown in Table 2 were prepared by performing a preparation process similar to the preparation method described in Example 2.

Example 3. Synthesis of 3- (1-allyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -N-hydroxy-propionamide (3h ')

Example Chemical structure NMR spectral data 3

6.89 (br t, 1H), 5.98-5.67 (m, 2H), 5.10-5.08 (m, 1H), 3.36 (t, J = 1.8 Hz, 2H), 2.61 (s, 2H), 2.06 (s, 2H ), 1.87 (s, 2 H)

Compounds of Examples 4 to 6 having physical properties as shown in Table 3 were prepared by carrying out a manufacturing process similar to the preparation method described in Reference Example 2.

Example 4. Synthesis of N-hydroxy-3- [1- (2-naphthalen-1-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (4n ')

Example 5 Synthesis of N-hydroxy-3- [1- (2-naphthalen-2-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (5n ')

Example 6 N-hydroxy-3- [2-oxo-1- (2-thiophen-2-yl-ethyl) -2,5-dihydro-1 H-pyrrol-3-yl] -propionamide Synthetic (6n ')

Example Chemical structure NMR spectral data 4

8.01 (d, J = 8.1 Hz, 1H), 7.77 (d, J = 8.7 Hz, 1H), 7.66 (d, J = 8.1 Hz, 1H), 7.48-7.20 (m, 4H), 6.62 (br t, 1H), 3.56 (s, 2H), 3.30-3.25 (m, 2H), 2.51 (t, J = 7.3 Hz, 2H), 2.25 (t, J = 7.3 Hz, 2H) 5

7.73-7.66 (m, 3H), 7.53 (s, 1H), 7.40-7.32 (m, 2H), 7.22 (s, 1H), 6.61 (br t, 1H), 3.69 (t, J = 7.3 Hz, 2H ), 3.60 (s, 2H), 2.97 (t, J = 7.0 Hz, 2H), 2.49 (t, J = 7.2 Hz, 2H), 2.24 (t, J = 7.3 Hz, 2H) 6

7.08 (br t, 1H), 6.85 (t, J = 4.0 Hz, 1H), 6.74 (s, 1H), 6.68 (br t, 1H), 3.65 (s, 4H), 3.37-3.29 (m, 1H) , 3.05 (t, J = 6.1 Hz, 2H), 2.51 (d, J = 4.8 Hz, 2H), 2.28 (s, 1H)

Compounds of Examples 7 to 8 having physical properties as shown in Table 4 were prepared by carrying out a manufacturing process similar to the preparation method described in Reference Example 3.

Example 7 3- [1- (3-Biphenyl-4-yl-propyl) -2-oxo-2,5-dihydro-1 H-pyrrol-3-yl] -N-hydroxy-propionamide Synthesis (7w ')

Example 8. Synthesis of N-hydroxy-3- [1- (3-naphthalen-2-yl-propyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (8w ')

Example Chemical structure NMR spectral data 7

7.51 (dd, J = 8.0 Hz, 4H), 7.37 (t, J = 7.4 Hz, 2H), 7.26 (q, J = 7.2 Hz, 3H), 6.81 (s, 1H), 4.79 (s, 2H), 3.89 (s, 2H), 3.48 (t, J = 7.1 Hz, 2H), 2.64 (t, J = 7.7 Hz, 2H), 2.56 (t, J = 5.1 Hz, 2H), 2.32 (t, J = 6.9 Hz, 1H) 8

10.54 (s, 1H), 7.71 (dd, J = 7.9 Hz, 3H), 7.54 (s, 1H), 7.41-7.33 (m, 2H), 7.24 (d, J = 7.8 Hz, 1H), 6.64 (s , 1H), 3.67 (s, 2H), 3.40 (s, 2H), 2.69 (t, J = 6.7 Hz, 2H), 2.57 (s, 2H), 2.41 (s, 2H), 1.86 (s, 2H)

Experimental Example 1. Pharmacological Activity Experiment

In order to search for the efficacy of the compounds of Examples 1 to 8 were carried out the following experiment.

1-1. HDAC Inhibition Test

HeLa cell nuclear extracts were used as histone deacetylase enzyme sources and histone dee of the compounds of the present invention using the HDAC Fluorescent Activity Assay / Drug Discovery Kit, Biomol, USA. Acetylase inhibitory activity was examined. This method is simpler than the method using a histone substrate labeled with radioactivity and has a very good sensitivity. Fluorogenic Histone Deacetylase Lysyl Substrate is used as a substrate for histone deacetylase. When the acetyl group is removed by histone deacetylase activity, the substrate is 360 It uses the principle of emitting light at an emission wavelength of 460 nm at an excitation wavelength of nm.

After reacting the histone deacetylase activity at 25 ° C. for 20 minutes in the presence of the compound of the present invention in each concentration (0.01-10 μM), the same amount of developer was added and 460 at an excitation wavelength of 350 nm. Emission wavelength of nm was measured by fluorescence spectrometer. IC ₅₀ values were defined as the concentration of test compound in which the fluorescence maximum was reduced by half.

The experimental results are shown in Table 5 below (AA means IC ₅₀ 's <1μM, A means IC ₅₀ 's <5μM, B means IC ₅₀ 's <10μM and C means IC ₅₀ 's> 10μM , IC ₅₀ 's means plural of IC ₅₀ )

Searching for histone deacetylase inhibitory activity as shown in Table 5, it was confirmed that the compounds of the present invention has excellent histone deacetylase inhibitory activity.

1-2. Tumor Cell Growth Inhibition Test

Human prostate tumor cell lines PC-3 or MDA-MB-231 (ATCC, USA) were cultured using RPMI 1640 medium containing 10% Fetal Bovine Serum (FBS). To measure anticancer activity, cells of appropriate concentration (approximately 5 x 10 ⁴ cells / mL) in RPMI 1640 medium containing 5% fetal bovine serum were dispensed into 96-well plates and then treated at 5% CO ₂ , 37 ° C. Incubated. After dispensing the cells, fix the cells by adding 50 μl of 50% trichloroacetic acid per well in a time zero (T ₀ ) plate to determine the cell concentration after one day and immediately before processing the compounds. It was set as. For cells treated with the compound, 50 μl of 50% trichloroacetic acid was added to the wells after 48 hours to fix the cells. Final concentrations of compounds added to the cells were adjusted to 0.01, 0.03, 0.1, 0.3, 1 μg / ml. The fixed plate was washed with tap water, dried, and stained with 100 μl / well of 0.4% solution of sulforhodamine B (SRB) dissolved in 0.1% acetic acid. After standing for 30 minutes, the resultant was washed with 0.1% acetic acid and dried at room temperature again, and then 10 mM tris base (pH 10.5) was added to dissolve the dyeing reagent. After absorbance measured at 540 nm was converted into a percentage of the control group, the concentration of the compound (IC ₅₀ (µg / ml)) that inhibits the growth of cancer cells by 50% was calculated.

To the experimental results shown in Table 5 (AA of the first column in the table, IC ₅₀ 's <1μM, A is IC _50' s <5μM, B are IC ₅₀ 's <10μM, and C are IC _50' s> 10 μM, and the second column is cell growth inhibitory activity compared to adriamycin, AA is equivalent (1-2 times), A is slightly weak (3-5 times), B is weak (5-10 times) And C means very weak (10 times or more).

As shown in Table 5 below, the effects of the compounds on the direct inhibition of HDAC and the growth inhibition of prostate tumor cells (PC-3) were determined. As a result, the compounds of the present invention were confirmed to have excellent growth inhibitory activity of cancer cells. .

Example HDAC Inhibitory Activity Cell growth inhibition (PC-3 or MDA-MB-231) One AA A 2 A AA 3 A C 4 _ - 5 - AA 6 C A 7 AA AA 8 AA AAA

Experimental Example 2. Acute Toxicity Test

Novel derivative compound of the present invention divided into 10 groups of 25 ± 5g ICR mice (Korean experimental animals) and 235 ± 10g SPF Sprague Dawley (Biogenomics) rats each divided into four groups Toxicity was observed for 24 hours after intraperitoneal administration of 600mg / kg, 3000mg / kg, 100mg / kg doses respectively.

As a result, no deaths were observed in all four groups, and no significant symptoms were found in weight gain and feed intake. Therefore, it was confirmed that the composition of the present invention has little acute toxicity.

The composition of the present invention can be administered in the following formulations, the following formulation examples are merely to illustrate the invention, whereby the content of the present invention is not limited.

Formulation Example 1 Preparation of Powder

Example 1 300 mg of a compound

Lactose 100 mg

Talc 10 mg

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2 Preparation of Tablet

Example 2 50 mg of compound

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 3 Preparation of Capsule

Example 3 50 mg of compound

Corn starch 100 mg

Lactose 100 mg

2 mg magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Formulation Example 4 Preparation of Injection

Example 4 50 mg of compound

Appropriate sterile distilled water for injection

pH adjuster

According to the conventional method for preparing an injection, the amount of the above ingredient is prepared per ampoule (2 ml).

Formulation Example 5 Preparation of Liquid

Example 5 Compound 1000 mg

20 g of sugar

20 g of isomerized sugar

Lemon flavor

Purified water was added to adjust the total volume to 1000 ml. According to the conventional method for preparing a liquid, the above components were mixed, and then filled into a brown bottle and sterilized to prepare a liquid.

The compound according to the present invention exhibits a strong growth inhibitory activity against tumor cells by using histone deacetylase as a functioning point, and the composition comprising the same can be used as a medicament for treating cancer diseases.

Claims (3)

A pharmaceutical composition for the treatment of cancer diseases comprising the compound of formula (I) or a pharmacologically acceptable salt thereof as an active ingredient:

(Ⅰ) Where X is -NHOH, n is an integer of 2, R is a C1-C4 alkyl or allyl group substituted with a hydrogen atom, naphthyl, thiophenyl or biphenyl, dotted line(

) Means a single bond or a double bond. delete The compound of claim 1 wherein: N-hydroxy-3- (1-naphthalen-2-yl-methyll-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (1e '), N-hydroxy-3- (1-methyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -propionamide (2h '), 3- (1-allyl-2-oxo-2,5-dihydro-1H-pyrrol-3-yl) -N-hydroxy-propionamide (3h '), N-hydroxy-3- [1- (2-naphthalen-1-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (4n '), N-hydroxy-3- [1- (2-naphthalen-2-yl-ethyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (5n '), N-hydroxy-3- [2-oxo-1- (2-thiophen-2-yl-ethyl) -2,5-dihydro-1H-pyrrol-3-yl] -propionamide (6n '), 3- [1- (3-biphenyl-4-yl-propyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -N-hydroxy-propionamide (7w '), Pharmaceuticals that are N-hydroxy-3- [1- (3-naphthalen-2-yl-propyl) -2-oxo-2,5-dihydro-1H-pyrrol-3-yl] -propionamide (8w ') Composition.