KR101958107B1 - Novel compounds, preparation method thereof and pharmaceutical composition for use in preventing or treating abnormal cell growth diseases containing the same as an active ingredient - Google Patents

Abstract

The present invention relates to a novel Lysine-specific demethylase 4A (KDM4A) inhibitory compound, a method for manufacturing the same, and a pharmaceutical composition for preventing or treating abnormal cell growth diseases containing the same as an active component. The novel KDM4A inhibitory compound according to the present invention can excellently inhibit the activity of KDM4A and proliferative activity of cancer cells, thereby being usefully used as the pharmaceutical composition containing the same as the active component for preventing or treating KDM4A-related diseases, for example, abnormal cell growth diseases, such as cancer.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel compound, a method for producing the same, and a pharmaceutical composition for preventing or treating abnormal cell growth diseases containing the same as an active ingredient. active ingredient}

The present invention relates to a novel compound, a process for producing the same, and a pharmaceutical composition for preventing or treating abnormal cell growth diseases containing the same as an active ingredient.

Lysine (K) -specific dimethylazine 4A (also known as KDM4A, JMJD2A) is an enzyme that performs histone modification, a key mechanism of post-hoc regulation. Histone methylation and dimerization (demethylation) occur in the lysine residues of histone proteins. Lysine residues can be single-, double- or tri- methylated and the reverse reaction is facilitated by a family of proteins called lysine (K) -specific dimethyla lase (KDMs).

Among these, KDM4A is classified as one of the Fe (II) oxygenase group because the enzyme reaction catalyzed by KDM4A uses divalent oxygen and Fe (II) ion. KDM4A can demethylate (demethylate) bi- and tri-methylated residues at lysine 9 and 36 positions of histone 3 in vitro (H3K9me3 / 2 and H3K36me3 / 2). However, in vivo in vivo, only the dimerization of the triplet methylated residue was observed. KDM4A has a greater affinity for H3K9me3 than H3K36me3. Although the H3K9 dimerization by KDM4A causes transcriptional activation of the promoter region, the functional effects of H3K36 dimetylation are unclear to date.

On the other hand, KDM4A appears to be associated with tumorigenesis in a variety of cancer types, and overexpression of KDM4A has been observed in prostate cancer, colon cancer, non-small cell lung cancer, triple negative breast cancer, and head and neck squamous cell carcinoma. In addition, overexpression and deficiency expression of KDM4A in bladder cancer have been reported to be related. The amplification and overexpression of KDM4A affects the number of replication times. The number of clones is one of the characteristics of cancer, and overexpression of KDM4A leads to increased replication of specific chromosomal regions. From the relationship between the carcinoma and KDM4A as described above, efforts for identifying a novel KDM4A inhibitor in cancer diseases have been continuously carried out.

On the other hand, KDM4A inhibitors can be divided into three groups according to the mode of action. First, the first group is a competitive inhibitor for α-KG (α-ketoglutarate), which is a cofactor for the KDM4A reaction that binds to Fe (II) molecules. All of the KDM4 subfamily enzymes require α-KG in the demethylation (demethylation) process, while 2,4-pyridinecarboxylic acid (PCDA) is a typical α-KG analog having KDM4A inhibitory activity.

The second group is a blocker of the metal cofactor (Zn ²⁺ ) required for catalysis. Disulfiram and ebselen inhibit KDM4A activity by releasing Zn ²⁺ ions from the Zn ²⁺ binding site of KDM4A (Non-Patent Document 2).

The third group consists of competitive inhibitors of the histone substrate. Analogs of methylated histone indicate the characteristics of KDM4A inhibitors.

However, in spite of efforts to continuously develop new KDM4A inhibitors as described above, there has not yet been reported development of such an inhibitor, and particularly, development of abnormal cell growth diseases, for example, cancer diseases, has not been significantly developed.

Accordingly, the present inventors have tried to develop a novel inhibitory compound that exhibits excellent inhibitory activity against KDM4A and at the same time has a significant effect on abnormal cell growth disease. The novel KDM4A inhibitory compound according to the present invention exhibits excellent inhibition of KDM4A activity, Cell proliferation activity can be excellently inhibited, and thus the present invention has been completed.

PloS one. 2010 Vol 5, Issue 11, p. e15535 R Sekirnik et al. Chem Commun (Camb) (42), 6376-6378. 2009 Sep 28.

It is an object of the present invention to provide a novel KDM4A (Lysine-specific demethylase 4A) inhibitory compound.

Another object of the present invention is to provide a method for producing the KDM4A inhibitory compound.

Still another object of the present invention is to provide a pharmaceutical composition for preventing or treating KDM4A (Lysine-specific demethylase 4A) -related disease containing the KDM4A inhibitory compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient .

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating abnormal cell growth diseases containing the KDM4A inhibitory compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a health functional food composition for preventing or ameliorating abnormal cell growth diseases containing the KDM4A inhibitory compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object,

The present invention provides a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

또는 치환 또는 비치환된 페닐이되,A is

Or substituted or unsubstituted phenyl,

Wherein said substituted phenyl is optionally substituted with one or more substituents independently selected from the group consisting of hydroxy, halogen, C _1-10 linear or branched alkoxy, and - (SO ₂ ) -C _1-10 straight or branched chain alkyl; May be substituted with more than one kind of substituent;

R ¹ , R ² , and R ³ are each independently H, hydroxy, C _1-3 straight chain or branched alkoxy.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound represented by the formula (2) with a compound represented by the formula (3) to prepare a compound represented by the formula (1).

[Reaction Scheme 1]

(In the above Reaction Scheme 1,

A, R ¹ , R ² , and R ³ are the same as defined in Formula 1,

Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating KDM4A (Lysine-specific demethylase 4A) -related disease containing the compound represented by the above-mentioned formula (1), a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient to provide.

The present invention also provides a pharmaceutical composition for preventing or treating abnormal cell growth diseases, which comprises the compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

Furthermore, the present invention provides a health functional food composition for preventing or ameliorating abnormal cell growth diseases, which comprises a compound represented by the above-mentioned formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

The novel KDM4A inhibitory compound according to the present invention can not only inhibit the activity of KDM4A (Lysine-specific demethylase 4A) but also can inhibit the proliferative activity of cancer cells to an excellent extent. Can be usefully used as a pharmaceutical composition for preventing or treating diseases related to KDM4A, for example, abnormal cell growth diseases, for example, cancer diseases.

1 is a graph showing the inhibitory activity (%) of KDM4A dimyrylase (demethylating enzyme) according to the treatment concentration of the compound of Example 11 of the present invention.
FIG. 2 is a diagram showing the compound of Example 11 of the present invention and the molecular docking at the KDM4A active site as an X-ray co-crystal structure using the Maestro program. FIG.
FIG. 3 shows the results of immunoblotting (protein analysis technique) showing changes in KDM4A, H3K9me1, H3K9me2, H3K9me3 and H3 in HCT-116 cells according to the treatment concentration of the compound of Example 11 of the present invention.
4 is a graph showing the proliferation activity (%) of HCT-116 cells according to compound treatment concentration at 2 days (48 hours) and 3 days (72 hours) after the treatment of the compound of Example 11 of the present invention.
FIG. 5 is a photograph showing the cell mobility of the compound of Example 11 of the present invention after scratching in HCT-116 cells, either untreated or treated (after 22 hours).
FIG. 6 shows the results of an experiment in which HCT-116 cells were grown in 10-cm plates (1 × 10 ⁶ cells / well) and treated with the compound of Example 11 at different concentrations or untreated (DMSO control group) (BD Biosciences, San Jose, CA, USA) after staining with Cell Apoptosis Kit and Alexa Flour 488 Annexin V (Life Technologies, Carlsbad, CA, USA) 116 cells of the present invention.
FIG. 7 is a graph showing the induction of PARP (Poly ADP ribose polymerase) degradation in HCT-116 cells by immunoblotting (protein analysis technique) after treating the compound of Example 11 with concentration or after treatment.

Hereinafter, the present invention will be described in detail as follows. It is to be understood, however, that the following description is presented to assist the understanding of the present invention, and the scope and spirit of the present invention is not limited to the following description.

The present invention provides a compound represented by the following formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof.

[Chemical Formula 1]

In Formula 1,

또는 치환 또는 비치환된 페닐이되,A is

Or substituted or unsubstituted phenyl,

Wherein said substituted phenyl is optionally substituted with one or more substituents independently selected from the group consisting of hydroxy, halogen, C _1-10 linear or branched alkoxy, and - (SO ₂ ) -C _1-10 straight or branched chain alkyl; May be substituted with more than one kind of substituent;

R ¹ , R ² , and R ³ are each independently H, hydroxy, C _1-3 straight chain or branched alkoxy.

In one aspect of the invention,

Wherein A is substituted or unsubstituted phenyl,

Wherein said substituted phenyl is selected from the group consisting of hydroxy, halogen, C _1-3 linear or branched alkoxy, and - (SO ₂ ) -C _1-3 linear or branched alkyl; And may be substituted with more than one kind of substituent.

In another aspect of the present invention,

Wherein A is substituted phenyl,

Wherein said substituted phenyl is optionally substituted with one or more substituents selected from the group consisting of hydroxy, halogen, C _1-3 linear or branched alkoxy, or - (SO ₂ ) -C _1-3 straight or branched chain alkyl Position) may be substituted phenyl.

In another aspect of the invention,

,

,

,

,

,

, 또는

일 수 있다.A is

,

,

,

,

,

, or

Lt; / RTI >

On the other hand, in the compound represented by the general formula (1)

In one aspect of the invention,

R ¹ , R ² , and R ³ are each independently H or hydroxy.

In another aspect of the present invention,

Wherein R ¹ , R ² , and R ³ are each independently H or hydroxy,

Wherein at least one of R ¹ , R ² , and R ³ is OH.

In another aspect of the invention,

^{Two of} R ¹ , R ² , and R ³ are hydroxy and the other is H,

Wherein the two hydroxides may be substituted at the 2 < 5 >

On the other hand, in the compound represented by the general formula (1)

The compound represented by the formula (1) is any one selected from the following group of compounds.

(1) (E) -tert-butyl 4- (2- (2,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(2) (E) -tert-butyl-2,2-dimethyl-4- (2- (2,3,4-trihydroxybenzylidene) hydrazinecarbonyl) -oxazolidin-3-carboxylate;

(3) (E) -tert-butyl 4- (2- (2,3-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(4) (E) -tert-butyl 4- (2- (2,4-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(5) (E) -tert-butyl 4- (2- (3,4-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(6) (E) -tert-butyl 4- (2- (3,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(7) (E) -tert-butyl 4- (2- (2-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(8) (E) -tert-butyl 4- (2- (3-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(9) (E) -tert-butyl 4- (2- (4-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(10) (E) -tert-butyl 4- (2-benzylidenehydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;

(11) (E) -N '- (2,5-dihydroxybenzylidene) benzohydrazide;

(12) (E) -N '- (2,5-dihydroxybenzylidene) -4-fluorobenzohydrazide;

(13) (E) -4-Chloro-N '- (2,5-dihydroxybenzylidene) benzohydrazide;

(14) (E) -4-Bromo-N '- (2,5-dihydroxybenzylidene) benzohydrazide;

(15) (E) -N '- (2,5-dihydroxybenzylidene) -4-hydroxybenzo hydrazide;

(16) (E) -N '- (2,5-dihydroxybenzylidene) -4-methoxybenzohydrazide; And

(17) (E) -N '- (2,5-dihydroxybenzylidene) -4- (methylsulfonyl) benzohydrazide.

In another aspect of the present invention,

The compound represented by the general formula (1)

(E) -N '- (2,5-dihydroxybenzylidene) -4-fluorobenzohydrazide;

(E) -4-chloro-N '- (2,5-dihydroxybenzylidene) benzo hydrazide;

(E) -4-bromo-N '- (2,5-dihydroxybenzylidene) benzo hydrazide;

(E) -N '- (2,5-dihydroxybenzylidene) -4-hydroxybenzo hydrazide;

(E) -N '- (2,5-dihydroxybenzylidene) -4-methoxybenzohydrazide; or

(E) -N '- (2,5-dihydroxybenzylidene) -4- (methylsulfonyl) benzohydrazide.

On the other hand, in one aspect of the present invention, the compound represented by Formula 1 may be used in the form of a pharmaceutically acceptable salt. Examples of the salt include pharmaceutically acceptable acid addition salts formed by free acid Salts are useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid and the like, aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, Derived from organic acids such as acetic acid, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid and the like. Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, But are not limited to, but are not limited to, but are not limited to, but are not limited to, but are not limited to, halides, halides, halides, halides, halides, halides, But are not limited to, lactose, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene Sulfonates, methanesulfonates, propanesulfonates, naphthalene-1-sulfonates, and the like, as well as sulfonates such as benzyl sulfonate, sulfonate, xylene sulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, -Sulfonate, naphthalene-2-sulfonate, mandelate, and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, by dissolving a derivative of the formula (1) in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile and the like, Followed by filtration and drying. Alternatively, the solvent and excess acid may be distilled off under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. In addition, the corresponding salt is obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

Furthermore, the present invention includes all the solvates, stereoisomers, hydrates, and the like, which can be prepared therefrom, as well as the compound represented by Formula 1 and pharmaceutically acceptable salts thereof.

The present invention also relates to a process for producing a compound represented by the formula (1)

Reacting a compound represented by the formula (2) with a compound represented by the formula (3) to prepare a compound represented by the formula (1).

[Reaction Scheme 1]

(In the above Reaction Scheme 1,

A, R ¹ , R ² , and R ³ are the same as defined in Formula 1,

Hereinafter, the method for preparing the compound represented by the formula (1) according to the present invention will be described in detail.

In one aspect of the method for preparing the compound represented by Chemical Formula 1 of Reaction Scheme 1 according to the present invention, the method may further include a step of preparing a compound represented by Chemical Formula 2 before the step represented by Reaction Scheme 1.

Here, the method of preparing the compound represented by Formula 3 may be understood as a step represented by the following Reaction Formula 2, for example.

[Reaction Scheme 2]

(2) is a step of reacting a compound represented by the formula (4) with hydrazine hydrate to prepare a compound represented by the formula (3).

Here, the hydrazine hydrate may be used in an excessive amount.

The solvent usable in Step 2 of the above reaction scheme is not particularly limited. Examples of the solvent include dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydrofuran (THF), methylene chloride, toluene, acetonitrile And a mixed solution thereof can be used, and methanol can be preferably used.

On the other hand, the reaction temperature in step 2 of the above reaction scheme is not limited as long as the product of formula (3) can be prepared, and can be carried out in consideration of the state of the solvent at, for example, -70 to -30 ° C , As another example, at -40 < 0 > C.

On the other hand, the reaction time in the second step of the above reaction scheme is included in the present invention without limitation as long as the product of formula (3) can be prepared. For example, the reaction time may be 5 to 24 hours, and in another example, 12 hours.

In one aspect of the present invention, a compound represented by formula (1) is reacted with a compound represented by formula (2) to obtain a compound represented by formula Is a step for preparing a compound to be displayed.

Here, the step represented by the above-mentioned Reaction Scheme 1 can be easily carried out by a person skilled in the art, which is included in the present invention without limitation, if the compound represented by Formula 1 can be prepared, Any method that can be changed or modified is included in the present invention without limitation.

Examples of the solvent that can be used in step 1 of the above reaction scheme include, but are not limited to, dimethylformamide (DMF), H ₂ O, methanol, ethanol, tetrahydrofuran (THF), methylene chloride, toluene, acetonitrile And a mixed solution thereof can be used, and anhydrous ethanol can be preferably used.

On the other hand, the reaction temperature in step 1 of the reaction scheme is included in the present invention without limitation as long as the product of formula (1) can be produced. For example, the reaction can be carried out at room temperature to 70 ° C, In another example, it may be carried out at 40 占 폚.

On the other hand, the reaction time in the above reaction scheme 1 is included in the present invention without limitation as long as it is the time for which the formula (1), which is the product, can be prepared, and is included in the present invention for 5 to 24 hours, for example, 12 hours, .

In one aspect of the present invention, the method for preparing the compound represented by the formula (1) can be carried out with reference to what has been done in the embodiment 1-17 of the present invention, Method. For example, the conditions such as the yield of the product, the use of the catalyst, the reaction temperature, and the time may be appropriately changed, or the order of the step may be changed, further steps may be further included, It is to be understood that the invention is not limited thereto.

Furthermore, the present invention relates to a pharmaceutical composition for preventing or treating KDM4A (Lysine-specific demethylase 4A) -related disease containing the compound represented by the above-mentioned formula (1), a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient to provide.

In one aspect of the invention,

Such KDM4A-related diseases may include all or some of the diseases caused directly or indirectly from abnormal conditions or symptoms such as abnormal activity, hyperactivity, overexpression, deficient activity, and deficient expression of KDM4A.

In another aspect of the present invention,

Such KDM4A-related diseases may be understood as referring to diseases that have been anticipated or identified as being related to KDM4A to date, for example, KDM4A-related diseases include abnormal cell growth diseases, Other examples include diseases related to KDM4A-related tumorigenesis, prostate cancer, colon cancer, non-small cell lung cancer, triple negative breast cancer and head and neck squamous cell carcinoma, . ≪ / RTI >

In another aspect of the invention,

The KDM4A-related disease is selected from the group consisting of lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, perianal cancer, gastric cancer, colon cancer, breast cancer, uterine cancer, Cancer of the prostate, chronic or acute leukemia, lymphocytic lymphoma, bladder cancer, cancer of the cervix, vaginal cancer, vulvar cancer, pozicin, esophagus cancer, small bowel cancer, endocrine cancer, thyroid cancer, pituitary cancer, adrenal cancer, soft tissue sarcoma, It is understood that the disease is one or more diseases selected from the group consisting of renal or ureter cancer, renal cell cancer, renal cancer, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, brain glioma glioma and pituitary adenoma,

In another aspect of the present invention, the abnormal cell growth disorder may be understood as being one or more diseases selected from the group consisting of psoriasis, benign prostatic hypertrophy, and retinopathy,

In another aspect of the present invention, the abnormal cell growth disorder may be understood to be a benign proliferative disease. Examples of the benign proliferative diseases include fibroadenoma, cirrhosis-related diseases, papilloma, and the like.

Meanwhile, the disease related to KDM4A (Lysine-specific demethylase 4A) may be understood to be a carcinoma or a malignant tumor, and the cancer is not particularly limited. Examples of the cancer include caustic myxoma, intrahepatic bile duct cancer, hepatoblastoma, liver cancer, Cancer, testicular cancer, myelodysplastic syndrome, glioblastoma, oral cancer, oral cancer, bacterial sarcoma, acute myelogenous leukemia, acute lymphocytic leukemia, basal cell carcinoma, ovarian epithelial cancer, ovarian germ cell cancer, male breast cancer, brain cancer, pituitary adenoma, multiple Cholangiocarcinoma, diffuse large B cell lymphoma, bladder cancer, bladder cancer, peritoneal cancer, pituitary cancer, adenocarcinoma, non-sinus carcinoma, cholangiocarcinoma, cholangiocarcinoma, cholangiocarcinoma, myeloma, gallbladder carcinoma, cholangiocarcinoma, colon cancer, chronic myelogenous leukemia, chronic lymphocytic leukemia, retinoblastoma, Small cell lung cancer, pediatric brain cancer, pediatric lymphoma, childhood leukemia, small bowel cancer, meningioma, esophageal cancer, glioma, neuroblastoma, renal cancer, kidney cancer, cardiac cancer, non-small cell lung cancer, non-Hodgkin's lymphoma, Cancer, duodenal cancer, malignant soft tissue tumor, malignant bone cancer, malignant lymphoma, malignant mesothelioma, malignant melanoma, ovary cancer, vulvar cancer, ureter cancer, urethral cancer, primary site unidentified cancer, gastric lymphoma, stomach cancer, gastric carcinoma, gastrointestinal stromal cancer Uterine cancer, uterine sarcoma, prostate cancer, metastatic cancer, metastatic brain cancer, mediastinal cancer, rectal cancer, rectal carcinoid, vaginal cancer, spinal cord cancer, breast cancer, sarcoma, Pancreatic cancer, salivary gland carcinoma, Kaposi sarcoma, Paget's disease, tonsil cancer, squamous cell carcinoma, lung cancer, lung cancer, lung squamous cell carcinoma, skin cancer, anal cancer, rhabdomyosarcoma, laryngeal cancer, pleural cancer, And the like.

Furthermore, preferably said cancer is acute myelogenous leukemia (AML); Chronic myelogenous leukemia (CML); Acute lymphocytic leukemia (ALL); Chronic lymphocytic leukemia (CLL); Hodgkin's disease (HD); Non-Hodgkin's lymphoma (NHL); B-cell lymphoma; T-cell lymphoma; Multiple myeloma (MM); Amyloidosis; Valgendrogmaglobulinemia; Myelodysplastic syndrome (MDS); Small lymphocytic lymphoma (SLL); Marginal lymphoma; Asymptomatic multiple myeloma; And bone marrow proliferative syndrome.

As used herein, the term "cancer" is used to establish unregulated or aberrantly regulated cell proliferation, reduced cellular differentiation, impaired ability to invade surrounding tissues, and / ≪ RTI ID = 0.0 > and / or < / RTI > The term "cancer" includes, but is not limited to, solid tumors and blood mediated tumors (hematologic malignant tumors). The term "cancer" includes diseases of the skin, tissues, organs, bone, cartilage, blood, and blood vessels. The term "cancer" further includes primary and metastatic cancers.

The solid tumor may be pancreatic cancer; Invasive bladder cancer containing bladder cancer; Colorectal cancer; Breast cancer including thyroid cancer, stomach cancer, metastatic breast cancer; Prostate cancer, including androgen-dependent and androgen-independent prostate cancer; Kidney cancer, including, for example, metastatic renal cell carcinoma; Liver cancer including hepatocellular carcinoma and intrahepatic bile duct; Lung and bronchial carcinoma, including non-small cell lung cancer (NSCLC), squamous cell lung cancer, bronchiolar lung carcinoma (BAC), adenocarcinoma of the lung, and small cell lung cancer (SCLC); Ovarian cancer, including, for example, advanced epithelial or primary peritoneal cancer; Cervical cancer; For example, uterine cancer including uterine body and cervix; Endometrial cancer; Gastric cancer; Esophagus cancer; For example, head and neck cancer including squamous cell carcinoma of the head and neck, nasopharyngeal cancer, oral cavity and pharynx; Melanoma; Neuroendocrine carcinoma, including metastatic neuroendocrine tumors; For example, brain tumors including glioma / glioblastomas, inverse dysplastic glomerulonephrosis, adult glioblastoma multiforme, and adult retroformed astrocytoma; Metastatic neuroendocrine tumors; Bone cancer; And neuroendocrine involvement including soft tissue sarcoma.

The hematological malignancies include acute myelogenous leukemia (AML); Chronic myelogenous leukemia (CML), including accelerated CML and CML aspirate (CML-BP); Acute lymphocytic leukemia (ALL); Chronic lymphocytic leukemia (CLL); Hodgkin's disease (HD); Non-Hodgkin's lymphoma (NHL), including follicular lymphoma and mantle cell lymphoma; B-cell lymphoma (including diffuse large B-cell lymphoma (DLBCL)); T-cell lymphoma; Multiple myeloma (MM); Amyloidosis; Valgendrogmaglobulinemia; (Including anemia of intractable anemia (RAE), hyperammonemia (RAEB), and transformation with accompanying RAEB (RAEB-T)) with mastitis formation syndrome (MDS) ; Small lymphocytic lymphoma (SLL); Marginal lymphoma; Asymptomatic multiple myeloma; And myeloproliferative syndrome.

In one aspect of the present invention, in the above-described KDM4A (Lysine-specific demethylase 4A) disease, the compounds of the present invention, its stereoisomers and acceptable salts thereof are useful for inhibiting the activity or expression of KDM4A (Lysine-specific demethylase 4A) And can be effectively used for the improvement, improvement, prevention or treatment of the above-mentioned diseases.

As used herein, the term "abnormal cell growth disorder" refers to a disorder characterized by uncontrolled or aberrant cell proliferation, reduced cellular differentiation, inappropriate ability to invade surrounding tissues, and / Which is characterized by the ability to induce cell proliferation. The term "abnormal cell growth disorder" includes, but is not limited to, solid tumors and blood mediated tumors (hematologic malignancies). It also includes abnormal cell growth disorders of the skin, tissues, organs, bone, cartilage, blood, and blood vessels. In addition, it includes primary and metastatic cancers.

In another aspect of the present invention, the compound represented by the formula (1), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof contained in the active ingredient of the pharmaceutical composition may be administered orally or parenterally When formulated, it can be prepared using a diluent or excipient such as a commonly used filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant and the like.

Examples of formulations for oral administration include tablets, pills, light / soft capsules, liquids, suspensions, emulsions, syrups, granules, elixirs and troches, , Dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine), lubricants (such as silica, talc, stearic acid and its magnesium or calcium salts and / or polyethylene glycols). The tablets may contain binders such as magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidine and may optionally contain binders such as starch, agar, alginic acid or sodium salts thereof Release or boiling mixture and / or absorbent, colorant, flavor, and sweetening agent.

The pharmaceutical composition containing the compound represented by Formula 1 as an active ingredient can be administered parenterally, and parenteral administration is performed by subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

In this case, in order to formulate the composition for parenteral administration, the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof may be mixed with water or a stabilizer or a buffer to prepare a solution or suspension, . The compositions may contain sterilized and / or preservatives, stabilizers, wettable or emulsifying accelerators, adjuvants such as salts and / or buffers for the control of osmotic pressure, and other therapeutically useful substances, Or may be formulated according to the coating method.

Further, the dose of the compound of the present invention represented by the formula (1) or a pharmaceutically acceptable salt thereof of the present invention to the human body may be varied depending on the age, weight, sex, dosage form, health condition and disease severity of the subject, Day is generally 0.1-1000 mg / day, preferably 1-500 mg / day, based on an adult subject of 70 kg, and may be divided once or several times a day at regular intervals according to the judgment of a doctor or pharmacist .

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, hormone therapy, chemotherapy and biological response modifiers for the prevention or treatment of diseases.

Further, the present invention relates to a health functional food composition for preventing or ameliorating a disease associated with KDM4A (Lysine-specific demethylase 4A) containing the compound represented by the above-mentioned formula (1), a stereoisomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient .

In one aspect of the present invention, it is understood that the KDM4A (Lysine-specific demethylase 4A) -related disease is a disease caused directly or indirectly from a state or phenomenon such as abnormal activity, hyperactivity, overexpression, etc. of KDM4A, As in the background of the present invention, can be understood as referring to diseases that have been anticipated or identified to be related to KDM4A to date, for example, KDM4A-related diseases can be understood as abnormal cell growth disorders, cancer diseases .

Meanwhile, KDM4A-related diseases of the health functional food composition are as described in the above pharmaceutical composition.

The present invention also relates to a method for the treatment and / or prophylaxis of KDM4A (Lysine) comprising administering to a subject in need thereof a therapeutically effective amount of a compound represented by Formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, -specific demethylase 4A) related diseases.

At this time, the disease related to KDM4A (Lysine-specific demethylase 4A) is as described in detail herein.

The therapeutically effective amount refers to a minimum amount that can improve symptoms or conditions of a subject when administered into the body, depending on the administration method.

Herein, the therapeutically effective amount may be different depending on the body weight, age, sex, condition, and family history of the subject to be administered. In the present invention, the treatment method can set different doses depending on different conditions .

In one aspect of the present invention, the dose of the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof to the human body may be varied depending on the age, weight, sex, dosage form, , Generally 0.1-1000 mg / day, preferably 1-500 mg / day, based on an adult subject having a body weight of 70 kg, and may be administered once a day at a predetermined time interval according to the judgment of a doctor or pharmacist It may be divided into several doses.

In another aspect of the invention, the "effective amount" may be an amount effective to treat proliferative, inflammatory, infectious, neurological, or cardiovascular disorders, or an amount effective to treat cancer.

In another aspect of the present invention, a compound according to the above method, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition or health functional food composition according to the present invention may further comprise any amount And any route of administration. The exact amount required will vary by subject depending upon the species, age, and general condition of the subject, the severity of the infection, the particular agent, its mode of administration, and the like. The compounds of the present invention are frequently formulated in dosage unit form for ease of administration and uniformity of dosage. The expression "dosage unit form" means a physically discrete unit of formulation suitable for the subject to be treated, as used herein. However, it will be understood that the total daily usage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific effective dosage level for any particular subject or organism will depend upon a variety of factors including:

In one example, the various factors include the severity of the disease and disorder to be treated; The activity of the specific compound employed; Specific composition; Age, weight, general health, gender and diet of the subject; The time of administration, the route of administration, and the rate of excretion of the particular compound employed; Duration of treatment; Drugs used in conjunction with or concurrently with the particular compound employed, and factors well known in the medical arts, for example.

On the other hand, the term "subject" as used herein can be understood to mean an organism, such as a mammal, a non-limiting animal such as a livestock, in one aspect, for example a human.

In one aspect of the invention, the pharmaceutical composition is administered orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (including powders, ointments, Lotion, lozenges, or drops), as an oral or nasal spray, and the like. In one particular embodiment of the invention, the compound of formula 1, its stereoisomer, or a pharmaceutically acceptable salt thereof is administered at a dose of about 0.01 mg / kg to about 50 mg / kg, for example, For example, from about 1 mg / kg to about 25 mg / kg of body weight per day, more than once per day, orally, parenterally.

Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents of the following examples commonly used in the art: water or other solvents, solubilizing agents, and emulsifying agents such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate , Benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (for example cottonseed, peanut, corn, germ, olive, castor, and pine oil), glycerol, Fatty acid esters of furfuryl alcohol, polyethylene glycol and sorbitan, and mixtures thereof. In addition to inert diluents, the oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and perfuming agents.

Injectable preparations, for example, sterile injectable aqueous or lipid productive suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. And isotonic sodium chloride solution. In addition, sterilized, fixed oils are conventionally used as a solvent or dispersion medium. For this purpose, any blend stationary oil may be utilized, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by incorporation of the active agent in the form of sterile solid compositions that are filtered or dissolved prior to use in a sterile water or other sterile injectable medium prior to use, have.

In order to sustain the effect of the compounds of the invention, slow absorption of the compound from subcutaneous or intramuscular injection is often desired. This can be achieved by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The absorption rate of the compound depends on the crystal size and its dissolution rate which may depend on the crystal form. Alternatively, delayed absorption of the parenterally administered compound form is achieved by dissolving or suspending the compound in an oil vehicle. The injectable depot form is made by forming a microencapsule matrix of the compound in a biodegradable polymer such as polylactide-polyglycolide. Depending on the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly (orthoesters) and poly (anhydrides). Depot injectable formulations are also prepared by entrapping the compound in liposomes or microemulsions compatible with body tissues.

Compositions for rectal or vaginal administration are, for example, suppositories which may be prepared by admixing the compounds of the present invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycols or suppository waxes, But is liquid at body temperature and therefore melts in the rectum or in the vagina to release the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such a solid dosage form, the active compound is mixed with: at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and / or a) filler or extenders such as starch, lactose C) humectant, such as glycerol, d) disintegrating agents, such as agar-agar-agar, for example, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; E) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate Sites, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid compositions of a similar type may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or lactose as well as high molecular weight polyethylene glycols and the like. Tablets, dragees, capsules, pellets, and granules in solid dosage form can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation arts. May optionally contain opacifying agents and may also be a composition that releases only the active ingredient (s), e.g., in a particular part of the intestinal tract, optionally in a delayed manner. Examples of embolic compositions that may be used include polymeric substrons and waxes. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or lactose as well as high molecular weight polyethylene glycols and the like.

In one aspect of the invention, the compound of formula 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof may also be in micro-encapsulated form with one or more excipients and one or more excipients as described above. have. Tablets, dragees, capsules, pellets, and granules in solid dosage forms may be prepared with coatings and shells such as enteric coatings, release modifying coatings and other coatings well known in the art of pharmaceutical formulation. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain additional substances other than inert diluents, such as tabletting lubricants and other tableting aids such as magnesium stearate and microcrystalline cellulose, as in normal practice. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents. May optionally contain opacifying agents and may also be a composition that releases only the active ingredient (s), e.g., in a particular part of the intestinal tract, optionally in a delayed manner. Examples of embolic compositions that may be used include polymeric substrons and waxes.

In another aspect of the invention, the topical or transdermal dosage form comprises ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed with the pharmaceutically acceptable carrier and any necessary preservatives or buffers under sterile conditions, as required. Ophthalmic formulations, ear drops, and eye drops are also contemplated to be within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches with the added benefit of providing controlled cleavage of the compound to the body. Such dosage forms can be made by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound across the skin. The speed can be controlled by providing a rate controlling membrane or by dispersing the compound in a polymer matrix or gel.

In some embodiments, a compound of the invention or a pharmaceutical composition thereof is administered with an anti-cancer agent. As used herein, the term "anti-cancer agent" refers to any agent that is administered to a subject having cancer to treat cancer. Combination therapy includes administration of a therapeutic agent either concurrently or sequentially. Alternatively, the therapeutic agent can be combined into one composition to be administered to a subject.

In one embodiment, the compounds of the invention may be used in combination with other therapeutic agents. In another embodiment, a compound of the invention is administered with a therapeutic agent selected from the group consisting of a cytotoxic drug, radiation therapy, and immunotherapy. In another embodiment, the compounds of the invention may be used in conjunction with chemotherapeutic regimens for the treatment of relapsed / intractable non-Hodgkin lymphoma, including DLBCL and CLL. Chemotherapeutic regimens include, but are not limited to, R-ICE (rituximab, ifosfamide, carboplatin and etoposide), R-DHAP (rituximab, dexamethasone, And cisplatin), and R-GDP (rituximab, gemcitabine, cisplatin and dexamethasone). It is to be understood that other combinations can also be embodied within the scope of the present invention.

Additional agents may be administered separately from the concomitant therapy provided as part of a multiple dose regimen. Alternatively, formulations may be part of a single dosage form mixed with a compound of the present invention. If administered as part of a combination therapy, the two therapeutic agents may be administered simultaneously, sequentially, or intermittently. Combination therapies may be used for any of the therapeutic indications described herein. In one embodiment, the combination therapy is for treating a proliferative disorder in a subject. In another embodiment, the proliferative disorder is breast cancer, lung cancer, ovarian cancer, multiple myeloma, acute myelogenous leukemia or acute lymphoblastic leukemia.

Another aspect of the invention relates to inhibiting KDM4A (Lysine-specific demethylase 4A) activity in a biological sample or a subject, said method comprising administering to a subject in need thereof a compound represented by formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof A salt, or a composition comprising said compound, or contacting said biological sample with said composition.

On the other hand, the term "biological sample " generally includes in vivo, in vitro, and in vitro materials, and also includes, but is not limited to, cell cultures or extracts thereof; A biopsied material obtained from a mammal or an extract thereof; And includes blood, saliva, urine, feces, semen, tears, or other body fluids or extracts thereof.

Another aspect of the present invention is to provide a kit comprising a separate container in a single package, wherein the compound or pharmaceutical composition and / or its salts disclosed herein act as a lysine-specific demethylase 4A (KDM4A) And one or more pharmaceutically acceptable carriers for use in the treatment of one or more disorders, symptoms and disorders.

Furthermore, the compound according to the present invention, its stereoisomer, or a pharmaceutically acceptable salt thereof, is effective for inhibiting the activity of KDM4A (Lysine-specific demethylase 4A) and thus improving or preventing abnormal cell growth diseases Was confirmed by the following experiment.

Specifically, as one specific example of the compound represented by the formula (1) of the present invention, the compound of Example 1-17 was prepared, and the inhibitory activity of KDM4A (Lysine-specific demethylase 4A) was tested on the enzyme unit to show excellent inhibitory activity (See Experimental Example 1 below), and further, the cancer cell line was observed to inhibit proliferation of the subject (see Experimental Example 2).

Particularly, from the experimental data of KDM4A inhibitory activity of Example 1-10, among the compounds of the present invention, the compound of Example 1, which is substituted with hydroxy at positions 2 and 5 of the left phenyl moiety of formula 1, Activity was observed. Further, in the case where 2,5-dihydroxyphenyl is used as in Example 1 and the A portion of Formula 1 is substituted or unsubstituted phenyl, that is, KDM4A which is more prominent in the case of Examples 11-17 Inhibitory activity was confirmed. Furthermore, in the above Examples 11-17, the substituent which facilitates hydrogen bonding, such as the case where the A is substituted phenyl and the 4-position of the phenyl is substituted by fluoro, hydroxy, methoxy, methylsulfonyl When it came, more excellent KDM4A inhibitory activity was confirmed.

Meanwhile, in order to confirm the effect of the compound represented by the formula (1) on the KDM4A activity in cells, the dimerization inhibitory activity was evaluated in HCT-116 human colorectal cancer cells using the compound of Example 11.

Invention Example 11 The compound is treated for 4 hours in HCT-116 cells and the methylation (methylation) level of H3K9 or H3K36 of KDM4A is then measured using antibodies against triplet methylation, double methylation, single methylation and total histone H3 Respectively.

As a result, it was confirmed that the single methylated histone H3 (H3K9me1) was dose-dependently reduced after the treatment with the compound of Example 11, and the level of triplomethylated H3K36 (H3K36me3) was ascertained (see FIG.

Accordingly, the compound represented by the formula (I) inhibits the activity of KDM4A, and thus the dimerization (demethylation) of the histone residue is excellently inhibited, and the pharmaceutical composition of the abnormal cell growth disease or the health functional food It could be used as an active ingredient of the composition.

On the other hand, as a result of testing on the effect of the compound of Example 11 on the HCT-116 cell proliferation, the number of living HCT-116 colon cancer cells after 48 hours and 72 hours of the compound treatment of Example 11 of the present invention, 11 compound. In particular, it was confirmed that when Example 11 was treated at a concentration of 50 μM, cell proliferation was completely inhibited (see FIG. 4).

On the other hand, it was confirmed that HCT-116 cell colony formation was also reduced in a dose-dependent manner in Example 11 compound treatment.

On the other hand, it was confirmed that the migration of HCT-116 cells was also inhibited by the compound treatment of Example 11.

Specifically, HCT-116 cells on the plate were subjected to scratching, and 24 hours of the change in the area of scratch reduction over time was examined. As a result, , It was confirmed that scratches disappeared, and under the treatment of Example 11, cell migration was inhibited in a dose-dependent manner.

On the other hand, the effect of Example 11 on apoptosis was examined in order to confirm the mechanism of cell proliferation inhibition. HCT-116 cells treated with Example 11 were stained with Annexin V and propidium iodide and flow cytometry was performed.

As a result, it was confirmed that the initial population of apoptosis was increased from 4.7% (DMSO control) to 22.3% (Example 11, 100 μM) by the treatment of Example 11. It was also confirmed that PARP cleavage was induced by treatment of Example 11 in HCT-116 cells.

In one aspect of the invention, epigenetic regulatory steps such as histone methylation / demethylation have been studied as therapeutic targets in the epigenetic approach to cancer therapy. Histone dimethylanase (demethylase) is a typical therapeutic target for epigenetic genetics, and KDM4A is prominent in oncology.

In recent years, the development of KDM4A inhibitors has become more demanded as the role of KDM4A in cancer development is clarified, as described above. The cell-based inhibitory activity, cancer cell proliferation or growth inhibition and cancer cell killing effect, which have been experimentally demonstrated in the present invention, are very important results from the above-mentioned viewpoints. In particular, the compound represented by the formula 1, , Or a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the same, or a health functional food composition will be useful for abnormal cell growth diseases, for example, cancer diseases.

In one aspect of the present invention, the introduction of 2,5-dihydroxy-substituted phenyl of the compound represented by Formula 1 and A substituted with hydroxy, halogen, methylsulfonyl or the like at the 4-position is more excellent in KDM4A May be considered essential for inhibitory activity.

In support of this, in silico molecular docking studies on KDM4A, the 2,5-dihydroxy-substituted moiety by the hydrogen bond and the π-cation interactions is the binding site amino acid (Asp135, Asp191, Lys241) and And the interaction was confirmed.

In another aspect of the present invention, the compound represented by formula (I) of the present invention has inhibitory activity on enzyme activity, inhibitory activity in cell-based system, and anti-tumor activity such as induction of cancer cell apoptosis.

Hereinafter, the present invention has been described in detail by way of examples and experimental examples.

However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the present invention is not limited to the following Examples and Experimental Examples.

Example 1 Preparation of (E) -tert-butyl 4- (2- (2,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidine-3-carboxylate

Step 1: Preparation of tert-butyl 4- (hydrazinecarbonyl) -2,2-dimethyl-oxazolidin-3-carboxylate

Dicarboxylate (500 mg) and hydrazine hydrate (excess) in methanol (30 mL) at -40 < 0 > C for 12 hours Lt; / RTI > The methanol was then removed under reduced pressure, the product was extracted with dichloromethane (2 X 500 mL), and the solution was dried over anhydrous sodium sulfate. The solvent was removed and the product was purified by flash chromatography column on silica gel (chloroform / MeOH = 20/1) (eluent containing 3% methanol in chloroform). The final product was obtained as an oil solution (442 mg, 85% yield).

^{1 H NMR (400 MHz, DMSO} -d 6) δ ppm 1.20 - 1.63 (m, 15 H) 3.63 - 3.91 (m, 1 H) 3.91 - 4.19 (m, 2 H) 4.19 - 4.34 (m, 2 H) 8.96 - 9.24 (m, 1H) ESI [M + H] = 260.2

Step 2: Preparation of (E) -tert-butyl 4- (2- (2,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

(1 eq., 260 mg) and 2,5-dihydroxy-benzaldehyde (1.5 eq., 205 mg) were added to anhydrous ethanol (10 mL) . After confirming the completion of the reaction by TLC, the ethanol was removed under reduced pressure, and the product was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to give the final product as a yellow solid (88%).

¹ H NMR (400 MHz, DMSO-d ₆ )? 1.24-1.48 (m, 12 H) 1.53 (s, 3 H) 3.68-3.95 (m, 1 H) 4.03-4.19 (m, 1H) 6.50-6.74 (m, 2H) 6.85-6.98 (m, 1H) 7.93-8.22 , 1 H) 11.62 (s, 1H) ESI [M + H] = 380.2

Example 2 Synthesis of (E) -tert-butyl-2,2-dimethyl-4- (2- (2,3,4-trihydroxybenzylidene) hydrazinecarbonyl) -oxazolidin- Manufacturing

The procedure of Example 1 was repeated except that 2,3,4-trihydroxy-benzaldehyde was used in place of the 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1, (Yield: 82%).

¹ H NMR (400 MHz, CHLOROFORM-d ₃ )? 1.45-1.75 (m, 15 H) 4.17 (s, 1H) (S, 1H), 6.50 (d, J = 8.24 Hz, 1H) 6.65 (d, J = 8.24 Hz, 1H) ] = 396.2

Example 3 Preparation of (E) -tert-butyl 4- (2- (2,3-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidine-3-carboxylate

The procedure of Example 1 was repeated except that 2,3-dihydroxy-benzaldehyde was used in place of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound (Yield: 85%).

¹ H NMR (400 MHz, DMSO- d ₆ )? 1.20-1.41 (m, 9 H) 1.44 (s, 3 H) 1.47-1.68 (m, 3 H) 3.77-3.95 (m, 1H) 4.20-4.39 (m, 1H) 6.57-6.87 (m, 2H) 6.93 (d, J = 7.79 Hz, 1H) 7.00-7.21 , 1 H) 9.23 (br s, 1H) 11.61-11.85 (m, 1H) ESI [M + H] = 380.2

Example 4 Preparation of (E) -tert-butyl 4- (2- (2,4-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidine-3-carboxylate

The procedure of Example 1 was repeated except that 2,4-dihydroxy-benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound (Yield: 89%).

¹ H NMR (400 MHz, DMSO- d ₆ )? 1.19-1.49 (m, 9 H) 1.50-1.57 (m, 3 H) 3.76-3.92 (m, 3 H) 4.05-4.18 1H), 7.93-8.16 (m, 1H), 9.85 (br s, 1H), 7.93-8.16 (m, 2H), 7.27 (dd, J = 8.47, 2.75 Hz, 1H) ESI [M + H] = 380.2 (m, < RTI ID = 0.0 >

Example 5 Preparation of (E) -tert-butyl 4- (2- (3,4-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

The procedure of Example 1 was repeated except that 3,4-dihydroxy-benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound (Yield: 92%).

¹ H NMR (400 MHz, DMSO- d ₆ )? 1.23-1.47 (m, 12 H) 1.53 (s, 3 H) 3.76 -3.91 , 7.21, 3.49 Hz, 1H) 6.58-7.07 (m, 3H) 7.61-7.85 (m, 1H) 9.12-9.41 [M + H] < / RTI > = 380.2

Example 6 Preparation of (E) -tert-butyl 4- (2- (3,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidine-3-carboxylate

Except that 3,5-dihydroxy-benzaldehyde was used in place of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound (Yield: 90%).

¹ H NMR (400 MHz, DMSO-d ₆ )? 1.23-1.47 (m, 12 H) 1.53 (s, 3 H) 3.76 - 3.91 (m, 1 H) 4.17 - (M, 1H) 7.07 (s, 1H) 7.61-7.85 (m, 1H) 9.12-9.41 (m, 1H) , 2 H) 11.20-11.39 (s, br, 1 H) ESI [M + H] = 380.2

EXAMPLE 7 Preparation of (E) -tert-butyl 4- (2- (2-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

The procedure of Example 1 was repeated except that 2-hydroxy-benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound Yield: 90%).

^{1 H NMR (300 MHz, CHLOROFORM-} d 3) δ 1.34-1.71 (m, 15 H) 3.98-4.22 (m, 2 H) 4.50 (d, J = 4.62 Hz, 1 H) 6.72-7.43 (m, 4 H) 8.25 (br s, 1H) 10.91 (s, 1H) ESI [M + H] = 364.2

Example 8 Preparation of (E) -tert-butyl 4- (2- (3-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

The procedure of Example 1 was repeated except that 3-hydroxy-benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to prepare the final desired compound Yield: 93%).

@ ¹ H NMR (300 MHz, CHLOROFORM-d ₃ ) .delta.1.27-1.45 (m, 3 H) 1.45-1.65 (m, 9H) 1.65-1.79 5.50 (m, 1H) 6.72-7.20 (m, 4H) ESI [M + H] = 364.2

Example 9 Preparation of (E) -tert-butyl 4- (2- (4-hydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

The procedure of Example 1 was repeated except that 4-hydroxy-benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound Yield: 88%).

¹ H NMR (300 MHz, CHLOROFORM-d ₃ )? 1.28-1.45 (m, 3 H) 1.45-1.65 (m, 19 H) 1.65-1.79 (m, 3 H) 4.05 (dd, J = 9.31, 2.56 Hz (M, 1H), 4.18-4.41 (m, 1H) 5.29 (dd, J = 7.09,2.64 Hz, 1H) 6.68-6.99 (m, 2H) 7.01-7.31 ] = 364.2

Example 10 Preparation of (E) -tert-butyl 4- (2-benzylidenehydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate

The procedure of Example 1 was repeated, except that benzaldehyde was used instead of 2,5-dihydroxy-benzaldehyde used in Step 2 of Example 1 to obtain the final desired compound (yield: 87%). .

¹ H NMR (300 MHz, CHLOROFORM- d ₃ )? 1.36-1.70 (m, 13 H) 1.75 (d, J = 9.23 Hz, 2 H) 3.94-4.19 (m, 1H) 4.27-4.42 H NMR (CDCl3) [delta] 7.38-7.51 (m, 3H) 7.51-7.68

Example 11 Preparation of (E) -N '- (2,5-dihydroxybenzylidene) benzo hydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and benzohydrazide (272.3 mg, 2.0 mmol) were stirred at 40 占 폚 overnight in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 85%).

^{1 H NMR (400 MHz, DMSO} -d 6) δ ppm 6.56 - 6.83 (m, 2 H) 6.98 (d, J = 2.44 Hz, 1 H) 7.46 - 7.76 (m, 3 H) 7.80 - 8.06 (m, 2 H) 8.57 (s, 1 H) 8.83 - 9.09 (m, 1 H) 10.24 - 10.52 (m, 1 H) 11.86 - 12.16 (m, 1H); MS (ESI): m / z = 431.43 [M + H].

< Example  12> Preparation of (E) -N '- (2,5-dihydroxybenzylidene) -4-fluorobenzohydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4-fluorobenzohydrazide (308.2 mg, 2.0 mmol) were stirred overnight at 40 ° C in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 85%).

¹ H NMR (400 MHz, DMSO- d ₆ )? Ppm 6.59-6.81 (m, 2 H) 6.99 (s, 1H) 7.29-7.55 s, 1H) 8.95 (s, 1H) 10.35 (s, 1H) 11.99 (s, 1H); MS (ESI): m / z = 275.13 [M + H].

Example 13 Preparation of (E) -4-chloro-N '- (2,5-dihydroxybenzylidene) benzohydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4-chlorobenzohydrazide (340.3 mg, 2.0 mmol) were stirred at 40 占 폚 overnight in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid, which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 80%).

^{1 H NMR (400 MHz, DMSO} -d 6) δ ppm 6.63 - 6.86 (m, 2 H) 7.00 (d, J = 2.14 Hz, 1 H) 7.76 (m, J = 8.55 Hz, 2 H) 7.88 (m , J = 8.24 Hz, 2 H) 8.57 (s, 1H) 8.98 (s, 1H) 10.31 (s, 1H) 12.04 (s, 1H); MS (ESI): m / z = 335.14 [M + H].

Example 14 Preparation of (E) -4-bromo-N '- (2,5-dihydroxybenzylidene) benzohydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4-bromobenzohydrazide (428.1 mg, 2.0 mmol) were stirred overnight at 40 ° C in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 83%).

¹ H NMR (400 MHz, DMSO-d ₆ )? Ppm 6.54 - 6.80 (m, 2 H) 6.99 (s, 1 H) 7.51 - 7.79 s, 1H) 8.97 (s, 1H) 10.24-10.40 (m, 1H) 12.02 (br s, 1H); MS (ESI): m / z = 291.12 [M + H].

Example 15 Preparation of (E) -N '- (2,5-dihydroxybenzylidene) -4-hydroxybenzo hydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4-hydroxybenzohydrazide (304.0 mg, 2.0 mmol) were stirred overnight at 40 ° C in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 85%).

¹ H NMR (400 MHz, DMSO- d ₆ )? Ppm 6.61-6.80 (m, 2 H) 6.80-6.97 (m, 2 H) 7.82 (s, 2 H) 8.52 (s, s, 1H) 10.15 (br s, 1H) 10.49 (br s, 1H) 11.80 (br s, 1H); MS (ESI): m / z = 273.23 [M + H].

Example 16 Preparation of (E) -N '- (2,5-dihydroxybenzylidene) -4-methoxybenzohydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4-methoxybenzohydrazide (332.1 mg, 2.0 mmol) were stirred overnight at 40 ° C in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 82%).

^{1 H NMR (400 MHz, DMSO} -d 6) δ ppm 3.84 (s, 3 H) 6.65 - 6.83 (m, 2 H) 6.95 (d, J = 2.44 Hz, 1 H) 7.07 (m, J = 8.85 Hz , 1H) 7.93 (m, J = 8.85 Hz, 2H) 8.54 (s, 1H) 8.97 (s, 1H) 10.46 (s, 1H) 11.88 (s, 1H); MS (ESI): m / z = 287.02 [M + H].

Example 17 Preparation of (E) -N '- (2,5-dihydroxybenzylidene) -4- (methylsulfonyl) benzohydrazide

2,5-dihydroxybenzaldehyde (462.3 mg, 3.0 mmol) and 4- (methylsulfonyl) benzohydrazide (428.3 mg, 2.0 mmol) were stirred overnight at 40 ° C in 5 ml of absolute ethanol. The solvent was then removed in vacuo and the crude product was obtained as a yellow solid, which was filtered and the residual benzaldehyde was washed with ethyl ether to give the final desired compound (yield: 80%).

The chemical structures of the final compounds prepared in Examples 1 to 17 are summarized in Table 1 below.

Example constitutional formula Example constitutional formula One

10

2

11

3

12

4

13

5

14

6

15

7

16

8

17

9

< Experimental Example  1> Evaluation of KDM4A inhibitory activity

In order to evaluate the inhibitory activity against the compound of the present invention against KDM4A, the following experiment was conducted.

Specifically, a homogeneous, time-resolved fluorescence (HTRF) assay was used to evaluate the KDM4A dimyrylase inhibitory activity of Example 1-17, and a recombinant protein containing KDM4A dimyrylase domain was analyzed by BPS Bioscience (San Diego, CA, USA). The optimum enzyme, Fe (NH ₄ ) ₂ (SO ₄ ) ₂ , L-ascorbic acid, α-ketoglutaric acid and substrate concentrations were determined by HTRF JMJD2A Histone H3K36 demethylation assay (Cisbio, France) Respectively.

(50 mM Tris-HCl, pH 7.5), 100 μM Fe (NH ₄ ) ₂ (SO ₄ ) ₂ , 200 μM L-ascorbic acid, 80 μM α-keto glutaric acid, 0.01% BSA To the reaction buffer containing 0.01% Tween 20, the KDM4A dimethylazaase was mixed with the compound of Example 1-17 and the peptide substrate diluted to 10 μM. After addition of the reagents for detection, the TR-FRET signal was measured with a Victor multilabel reader (Perkin Elmer, Waltham, MA, USA) to assess inhibitory activity. IC ₅₀ was calculated by nonlinear regression using Prism software (GraphPad, La Jolla, Calif., USA).

The data obtained from the above experiment are shown in Table 2 below.

Example KDMA4A inhibitory activity at 10 [mu] M
(%) KDMA4A IC ₅₀
(μM) One 57.4 31.4 2 36.3 - 3 8.75 - 4 -5.1 - 5 28.0 - 6 39.5 - 7 22.2 - 8 5.67 - 9 16.8 - 10 12.0 - 11 67.1 6.56 12 61.1 2.25 13 13.2 - 14 19.9 - 15 84.0 1.63 16 84.9 3.49 17 54.6 2.67

As shown in Table 2, the compounds of Examples 1-17 of the present invention were found to exhibit KDMA4A inhibitory activity at a concentration of 10 μM, and in particular, 2,5-dihydroxy substituted compounds of Example 1 and Examples 10-17 And exhibited remarkably excellent KDMA4A inhibitory activity even in the IC ₅₀ data value.

Therefore, the novel KDM4A (Lysine-specific demethylase 4A) inhibitor of the present invention, its stereoisomer, or its pharmaceutically acceptable salt can excellently inhibit KDMA4A activity, and KDM4A (Lysine -specific demethylase 4A) related diseases, for example, a pharmaceutical composition for preventing or treating abnormal cell growth diseases, or a health functional food composition.

&Lt; Experimental Example 2 > Evaluation of proliferation inhibitory activity in cancer cell lines

In order to evaluate the proliferation inhibitory activity of the compound of the present invention in the HCT-116 human colon cancer cell line, the following experiment was conducted.

Specifically, HCT-116 human colon cancer cells were purchased from ATCC (Rockville, MD, USA) and cell viability was evaluated by tetrazolium-based assay using EZ-Cytox Cell Viability Assay kit (DaeilLab, Korea) . Briefly, HCT-116 2000 cells were plated in 96-well plates in 100 [mu] l of medium. The following day, cells were treated with the compound of the invention example with DMSO (Fisher, Waltham, MA, USA) and used as a negative control. At 3 days (72 hours) after addition of the drug, 15 E of EZ-Cytox kit reagent was added to each well of a 96-well plate and incubated at 37 캜 for 4 hours in a humidified CO ₂ incubator. After incubation, the optical density (OD) was measured at 450 nm using a Victor multilabel reader (Perkin Elmer, Waltham, Mass., USA). GI ₅₀ was calculated by nonlinear regression using Prism version 5.01 (GraphPad, La Jolla, CA, USA).

Example HCT-116 human colon cancer cells GI ₅₀
(μM) 11 54.27 12 39.07 13 - 14 - 15 > 100 16 44.01 17 56.4

As shown in Table 3, the compounds of Examples 11-17 of the present invention exhibit excellent cancer cell line proliferation inhibitory activity against HCT-116 human colon cancer cells, and in particular, the compound of Example 12 exhibits the best cancer cell proliferation inhibitory activity can confirm.

Therefore, the novel KDM4A (Lysine-specific demethylase 4A) inhibitory compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof according to the present invention can excellently inhibit abnormal cell growth, for example, cancer cell proliferation, For example, a pharmaceutical composition for preventing or treating cancer, or a health functional food composition.

Claims (10)

Prostate cancer, non-small cell lung cancer, triple negative breast cancer, head and neck squamous cell carcinoma, or bladder cancer, comprising a compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof as an active ingredient: Therapeutic pharmaceutical compounds:
(1) (E) -tert-butyl 4- (2- (2,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;
(11) (E) -N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(12) (E) -N '- (2,5-dihydroxybenzylidene) -4-fluorobenzohydrazide;
(13) (E) -4-Chloro-N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(14) (E) -4-Bromo-N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(15) (E) -N '- (2,5-dihydroxybenzylidene) -4-hydroxybenzo hydrazide;
(16) (E) -N '- (2,5-dihydroxybenzylidene) -4-methoxybenzohydrazide; And
(17) (E) -N '- (2,5-dihydroxybenzylidene) -4- (methylsulfonyl) benzohydrazide.
delete delete delete delete delete The method according to claim 1,
Wherein said compound inhibits KDM4A (Lysine-specific demethylase 4A) activity.
The method according to claim 1,
Wherein said cancer is a cancer associated with KDM4A (Lysine-specific demethylase 4A).
delete Prostate cancer, non-small cell lung cancer, triple negative breast cancer, head and neck squamous cell carcinoma, or bladder cancer, comprising a compound selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof as an active ingredient: Health functional foods composition for improvement:
(1) (E) -tert-butyl 4- (2- (2,5-dihydroxybenzylidene) hydrazinecarbonyl) -2,2-dimethyloxazolidin-3-carboxylate;
(11) (E) -N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(12) (E) -N '- (2,5-dihydroxybenzylidene) -4-fluorobenzohydrazide;
(13) (E) -4-Chloro-N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(14) (E) -4-Bromo-N '- (2,5-dihydroxybenzylidene) benzohydrazide;
(15) (E) -N '- (2,5-dihydroxybenzylidene) -4-hydroxybenzo hydrazide;
(16) (E) -N '- (2,5-dihydroxybenzylidene) -4-methoxybenzohydrazide; And
(17) (E) -N '- (2,5-dihydroxybenzylidene) -4- (methylsulfonyl) benzohydrazide.

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