KR20170110017A - Benzenesulfonamide derivatives, preparation method thereof, and pharmaceutical composition for use in treating cancer and asthma - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for the treatment of cancer or asthma, comprising a benzenesulfonamide derivative represented by the general formula (1) as an active ingredient, wherein the compound represented by the general formula (1), its optical isomer or its pharmaceutically acceptable Possible salts are useful not only for the activity of Pin1 isomerase but also for inhibiting the cell proliferation of cancer cell lines, and thus can be useful as a pharmaceutical composition for preventing or treating cancer or chronic inflammatory diseases (such as asthma) associated therewith.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a benzenesulfonamide derivative, a preparation method thereof, and a pharmaceutical composition for treating cancer or asthma,

The present invention relates to a pharmaceutical composition for treating cancer or asthma comprising the benzenesulfonamide derivative represented by the formula (1) as an active ingredient.

Human cancer cells are diseases caused by abnormalities of various cancer signaling pathways such as AP1 (activator protein 1), NFkB, b-catenin, NFAT (nuclear factor of activated T-cells) The activation of the cancer-induced gene or the inactivation of the cancer-suppressing gene is provided as a means for treating cancer cells.

Recently, the discovery of Proryl isomerase, Pin1 (Protein Interacting with Nima), has been shown to play an important role in determining protein function following Pin1 mediated protein conformation following protein phosphorylation. It is known that phosphorylation of proteins serine, threonine and tyrosine residues occurs during the intracellular signal transduction. The Pin1 gene is the proline amino acid site behind the phosphorylated serine or threonine amino acid residue of a specific protein, ie, phosphorylated Ser / Thr-Pro To promote the cis / trans isomerization of proline residues, thereby changing the structure of the protein and regulating the functions of the intracellular proteins (Non-Patent Document 1).

Pin1 isozyme catalyzes the activation of 19 oncogenes including beta-catenin, AKT, cyclin D1, Plk, NF-kB, Stat3, Neu, Mcl1, c-Jun, c-Fos and c-Myb Raf- , And inhibits the activity of thirteen tumor suppressors such as p53, Bax, Bcl-1, and Fbw7 and plays an important role in cancer development as a whole. Thus, Pin1 protein has been used for the diagnosis and prognosis of cancer, As a target of the report.

The cancer treatment of the patient improves the survival rate of the patient through primary surgical operation and radiation therapy, but a new treatment alternative is needed to obtain an effective treatment for advanced cancer (Non-Patent Document 2) Chemotherapy and induction of apoptosis by low molecular weight substance transport have emerged as an alternative (Non-Patent Document 3).

In surgically resected specimens, Pin1 has been reported to be highly expressed in a variety of cancers including colon, cervical, breast, lung, and pancreatic cancer. Pin1 knockout mice exhibit early symptoms of senescence or some carcinogenic genes that inhibit cancer, while transgenic mice overexpressing Pin1 show a centromeric increase and a cancer-causing phenotype. In addition, specific inhibitor or siRNA can be used to induce mitotic arrest and apoptosis in Pin1 knockdown cancer cells. Thus, Pin1 isozyme inhibitors may be a novel therapeutic strategy and alternative, highlighted in the same Pin1-overexpressing carcinoma, such as cancer, and this type of inhibitor may be a realistic treatment alternative for advanced solid tumors.

Asthma is a type of chronic inflammatory disease that occurs in the airways. Mast cell, eosinophil, and T cell infiltration are involved and cause airway inflammation. Asthma has a high incidence rate in North and South America, South America, Europe and Oceania, and has been steadily increasing since the 1970s, causing 300 million patients every year and causing 250,000 deaths. Various antigens that enter the airway invade mast cells or Th2 cells, and cytokines such as Interleukin-4, GM-CSF, and Interleukin-5, which are expressed here, induce eosinophil from bone marrow, and eosinophil Thereby increasing the survival rate. This activates various cytokines and promotes the development of proteins such as granule protein, leukotriene, neurotoxin, peroxidase, and cationic protein and causes airway injury. Activated TGF-beta1 is also known to contribute to asthma by affecting airways by this mechanism.

Pin1 isozyme is activated in eosinophil in asthmatic patients and is known to stabilize mRNA of GM-CSF (Non-Patent Document 4) and regulate TGF-beta1 and Interleukin-5 (Non-Patent Document 5). Treatment of the Pin1 inhibitor resulted in decreased expression of GM-CSF and interleukin-5, reduced allergic inflammation induced by eosinophil, reduced eosinophil-induced pulmonary inflammation and expression of TGF-beta1 in rat- (Non-Patent Document 6). Thus, Pin1 isozyme inhibitors can be a new therapeutic strategy and alternative in asthma patients.

Accordingly, the inventors of the present invention have been studying a compound that inhibits the Pin1 isomerase, suggesting that the compound represented by the formula (1), its optical isomer or a pharmaceutically acceptable salt thereof according to the present invention has a markedly inhibitory effect on the activity of the Pin1 isomerase And thus can be useful as a preventive or therapeutic agent for cancer and asthma, and completed the present invention.

Lu PJ et al. Science. 1999, Feb 26; 283 (5406): 1325-8 Pikarsky E et al. Nature. 2004, 431: 461-466 Llovet JM et al. Hepatology. 2003, 37: 429-442 Shen ZJ et al. Nature Immunology. 2005, Dec 6 (12): 1280-1287 Esnault S et al. J Allergy Clin Immunol. 2007, Nov; 120 (5): 1082-1088 Shen ZJ et al. J Clin Invest. 2008, Feb; 118 (2): 470-490

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cancer or chronic inflammatory diseases comprising a compound represented by the formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for preventing or treating a cancer or chronic inflammatory disease comprising a compound represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

[Chemical Formula 1]

In Formula 1,

R ¹ is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, or C _1-10 straight or branched alkylaminocarbonyl;

R ² is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, C _1-10 straight or branched chain haloalkoxy, or C _1-10 straight or branched alkylthio ;

R ³ and R ⁴ are independently hydrogen, halogen, C _1-10 straight or branched chain alkyl, or C _1-10 straight or branched alkoxy;

Wherein R ¹ and R ² together with the carbon atoms to which they are attached may form a 5-7 membered heteroaryl comprising at least one heteroatom selected from the group consisting of N, O and S, Aryl may be substituted with at least one member selected from the group consisting of C _1-10 linear or branched alkyl, oxo, and C _1-10 straight chain or branched alkylcarbonyl;

R ³ and R ⁴ together with the carbon atoms to which they are attached may form a C _6-12 aryl.

Herein, the cancer may be cervical cancer, colon cancer, pancreatic cancer, ovarian cancer, cervical cancer, and the like. The chronic inflammatory disease may be asthma or the like.

The compound represented by the formula (I) according to the present invention, its optical isomer or pharmaceutically acceptable salt thereof is excellent in the activity to inhibit the cell proliferation of the cancer cell line as well as the activity of the Pin1 isomerase, (Asthma and the like) of the present invention.

Fig. 1 (a) is a graph showing the inhibition rate of cell growth after HeLa cells treated with the compound of Example 1 at different concentrations.
Fig. 1 (b) shows the expression of Pin1 protein in HeLa cells (normal Pin1 expression (shCon) or low expression (shPin1)).
1 (C) is a graph showing the apoptosis of HeLa cells (normal Pin1 expression (shCon) or low expression (shPin1)) treated with the compound of Example 1 at different concentrations.
FIG. 2 is a graph showing Pin1 protein expression level in pancreatic cancer cell line and cell growth inhibition rate after treatment of pancreatic cancer cell line according to concentration of compound of Example 1. FIG.
FIG. 3 is a graph showing the selective expression of Pin1 target protein after treating the cervical cancer cell line HeLa (normal Pin1 expression (shCon) or low expression (shPin1)) according to the concentration of the compound of Example 1.
FIG. 4 is a graph showing the binding of the compound of Example 6 to Pin1 protein by treating the compound according to concentration. FIG.
FIG. 5 is a graph showing the cell migration after treating the compound of Example 6 with the ATRA compound as a control group and BxPC3 as a pancreatic cancer cell line. FIG.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing or treating a cancer or chronic inflammatory disease comprising a compound represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient.

In Formula 1,

R ¹ is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, or C _1-10 straight or branched alkylaminocarbonyl;

R ² is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, C _1-10 straight or branched chain haloalkoxy, or C _1-10 straight or branched alkylthio ;

R ³ and R ⁴ are independently hydrogen, halogen, C _1-10 straight or branched chain alkyl, or C _1-10 straight or branched alkoxy;

Wherein R ¹ and R ² together with the carbon atoms to which they are attached may form a 5-7 membered heteroaryl comprising at least one heteroatom selected from the group consisting of N, O and S, Aryl may be substituted with at least one member selected from the group consisting of C _1-10 linear or branched alkyl, oxo, and C _1-10 straight chain or branched alkylcarbonyl;

R ³ and R ⁴ together with the carbon atoms to which they are attached may form a C _6-12 aryl.

Preferably,

Wherein R ¹ is hydrogen, halogen, C _1-4 straight or branched alkyl, C _1-4 straight or branched alkoxy, or C _1-4 straight or branched alkylaminocarbonyl;

R ² is hydrogen, halogen, C _1-4 straight or branched chain alkyl, C _1-4 straight or branched alkoxy, C _1-4 straight or branched chain haloalkoxy, or C _1-4 straight or branched alkylthio ;

R ³ and R ⁴ are independently hydrogen, halogen, straight or branched C _1-4 alkyl, or C _1- 4 straight or branched chain alkoxy of;

Wherein R ¹ and R ² together with the carbon atoms to which they are attached may form a 5-6 membered heteroaryl comprising at least one heteroatom selected from the group consisting of N, O and S, Aryl may be substituted with at least one member selected from the group consisting of C _1-4 straight or branched chain alkyl, oxo, and straight or branched C _1-4 alkylcarbonyl;

Wherein R ³ and R ⁴ may form a C _6-8 aryl group with the carbon atom to which they are connected.

More preferably,

Wherein R ¹ is hydrogen, F, Cl, methyl, ethyl, methoxy, ethoxy, or isopropylaminocarbonyl;

R ² is hydrogen, F, Cl, methyl, methoxy, ethoxy, trifluoromethoxy, or methylthio;

R ³ is hydrogen;

R ⁴ is hydrogen, Cl, methyl, methoxy or ethoxy, and;

Said R < ¹ > and R < ² > together with the carbon atoms to which they are attached may form a pyrrolidine substituted with morpholine or methylcarbonyl substituted with methyl and oxo groups;

Wherein R < ³ > and R < ⁴ > together with the carbon atoms to which they are attached may form a phenyl.

Preferred examples of the compound represented by the formula (1) according to the present invention include the following compounds, and the following compounds were purchased from ChemBridge Corporation (16981 Via Tazon, Suite G, San Diego, CA 92127, USA).

1) N- (3- (1H-imidazol-1-yl) propyl) -4-chloronaphthalene-1-sulfonamide;

2) N- (3- (1H-imidazol-1-yl) propyl) -4-methoxy-3-methylbenzenesulfonamide;

3) N- (3- (1H-imidazol-1-yl) propyl) -4,5-dichloro-2-methylbenzenesulfonamide;

4) N- (3- (1H-imidazol-1-yl) propyl) -4-fluoro-3-methylbenzenesulfonamide;

5) N- (3- (1H-imidazol-1-yl) propyl) -2,5-dichlorobenzenesulfonamide;

6) N- (3- (1H-imidazol-1-yl) propyl) -4-butoxybenzenesulfonamide;

7) N- (3- (1H-imidazol-1-yl) propyl) -5-fluoro-2-methoxybenzenesulfonamide;

8) N- (3- (1H-imidazol-1-yl) propyl) -2,5-diethoxy-4-methylbenzenesulfonamide;

9) N- (3- (1H-imidazol-1-yl) propyl) -2-methoxy-4,5-dimethylbenzenesulfonamide;

10) N- (3- (1H-imidazol-1-yl) propyl) -4- (trifluoromethoxy) benzenesulfonamide;

11) N- (3- (1H-imidazol-1-yl) propyl) -4-ethoxybenzenesulfonamide;

2-methyl-3-oxo-3,4-dihydro-2H-benzo [b] [1,4] oxazine- - sulfonamide;

13) N- (3- (1H-imidazol-1-yl) propyl) -3-fluoro-4-methoxybenzenesulfonamide;

14) N- (3- (1H-imidazol-1-yl) propyl) -3-ethyl-4-methoxybenzenesulfonamide;

15) N- (3- (1H-imidazol-1-yl) propyl) -5-chloro-2-methoxybenzenesulfonamide;

16) N- (3- (1H-imidazol-1-yl) propyl) -4-methoxynaphthalene-1-sulfonamide;

17) N- (3- (1H-imidazol-1-yl) propyl) -1-acetylindoline-5-sulfonamide;

18) 5- (N- (3- (1H-Imidazol-1-yl) propyl) sulfamoyl) -N-isopropyl-2-methylbenzamide;

19) N- (3- (1H-imidazol-1-yl) propyl) -4- (methylthio) benzenesulfonamide;

20) N- (3- (1H-imidazol-1-yl) propyl) -3-chloro-4-methoxybenzenesulfonamide;

21) N- (3- (1H-imidazol-1-yl) propyl) -3-ethoxy-4-fluorobenzenesulfonamide;

22) N- (3- (1H-imidazol-1-yl) propyl) -4-chloro-3-methoxybenzenesulfonamide;

23) N- (3- (1H-imidazol-1-yl) propyl) -4-methylbenzenesulfonamide; And

24) N- (3- (1H-Imidazol-1-yl) propyl) -3,4-dimethylbenzenesulfonamide.

The compound represented by the formula (1) of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is 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, or by distillation of the solvent and excess acid 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).

Further, a pharmaceutically acceptable salt can be prepared using an amino acid having an amino group attached to an organic acid. Examples of the amino acid salt include natural amino acids such as glycine, alanine, phenylalanine, valine, lysine and glutamic acid And most preferably L-lysine. ≪ Desc / Clms Page number 2 >

In addition, the present invention encompasses not only the compound represented by the formula (1) and pharmaceutically acceptable salts thereof, but also solvates, optical isomers and hydrates thereof which can be prepared therefrom.

In the pharmaceutical composition for preventing or treating cancer or chronic inflammatory disease of the present invention, the cancer may be cervical cancer, colon cancer, pancreatic cancer, ovarian cancer, cervical cancer, etc., and the chronic inflammatory disease may be asthma and the like.

The pharmaceutical composition according to the present invention is characterized by inhibiting the Pin1 isomerization enzyme.

The compound of formula (I) according to the present invention may be administered orally or parenterally in a variety of formulations at the time of clinical administration. In the case of formulation, the compound of the present invention may be used as a filler, an extender, a binder, a wetting agent, a disintegrant, Diluents or excipients.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules, troches, and the like, which may contain one or more excipients such as starch, calcium carbonate, Sucrose, lactose, gelatin or the like. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like are included in addition to commonly used simple diluents such as water and liquid paraffin. .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the non-aqueous solvent and suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

The effective dose of the compound of the present invention on the human body may vary depending on the age, weight, sex, dosage form, health condition and disease severity of the patient, and is generally about 0.001-100 mg / kg / 0.0 > mg / kg / day. ≪ / RTI > It is generally from 0.07 to 7000 mg / day, preferably from 0.7 to 2500 mg / day, based on adult patients weighing 70 kg, and may be administered once a day It may be divided into several doses.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

The compounds of Examples 1-24 below were purchased from ChemBridge Corporation (16981 Via Tazon, Suite G, San Diego, CA 92127, USA).

< Example  1> N- (3- (1H-imidazol-1-yl) propyl) -4-chloronaphthalen- Sulfonamide

< Example  2> N- (3- (1H-imidazol-1-yl) propyl) -4- Methoxy -3- Methylbenzenesulfonamide

< Example  3> N- (3- (1H-Imidazol-1-yl) propyl) -4,5- Dichloro -2- Methylbenzenesulfonamide

< Example  4> N- (3- (1H-Imidazol-1-yl) propyl) -4- Fluoro -3- Methylbenzenesulfonamide

Example 5 Synthesis of N- (3- (1H-imidazol-1-yl) propyl) -2,5-dichlorobenzenesulfonamide

Example 6 Synthesis of N- (3- (1H-imidazol-1-yl) propyl) -4-butoxybenzenesulfonamide

< Example  7> N- (3- (1H-Imidazol-1-yl) propyl) -5- Fluoro -2- Methoxybenzenesulfonamide

< Example  8> N- (3- (1H-imidazol-1-yl) propyl) Diethoxy -4- Methylbenzenesulfonamide

< Example  9> N- (3- (1H-Imidazol-1-yl) propyl) -2- Methoxy -4,5- Dimethylbenzenesulfonamide

< Example  10> N- (3- (1H-imidazol-1-yl) propyl) -4- ( Trifluoromethoxy ) Benzenesulfonamide

Example 11 Synthesis of N- (3- (1H-imidazol-1-yl) propyl) -4-ethoxybenzenesulfonamide

< Example  12> N- (3- (1H-Imidazol-1-yl) propyl) -2- methyl -3-oxo-3,4- Dihydro -2H-benzo [b] [1,4] oxazine-6-sulfonamide

< Example  13> N- (3- (1H-Imidazol-1-yl) propyl) -3- Fluoro -4- Methoxybenzenesulfonamide

< Example  14> N- (3- (1H-Imidazol-1-yl) propyl) Methoxybenzenesulfonamide

< Example  15> N- (3- (1H-Imidazol-1-yl) propyl) -5- Chloro -2- Methoxybenzenesulfonamide

< Example  16> N- (3- (1H-imidazol-1-yl) propyl) -4- Methoxynaphthalene -One- Sulfonamide

< Example  17> N- (3- (1H-Imidazol-1-yl) propyl) Acetylindoline -5- Sulfonamide

< Example  18> 5- (N- (3- (1H-imidazol-1-yl) propyl) Sulfamoyl ) -N-isopropyl-2-methylbenzamide

Example 19 Synthesis of N- (3- (1H-imidazol-1-yl) propyl) -4- (methylthio) benzenesulfonamide

< Example  20> N- (3- (1H-Imidazol-1-yl) propyl) -3- Chloro -4- Methoxybenzenesulfonamide

< Example  21> N- (3- (1H-Imidazol-1-yl) propyl) -3- Ethoxy -4- Fluorobenzenesulfonamide

< Example  22> N- (3- (1H-Imidazol-1-yl) propyl) -4- Chloro -3- Methoxybenzenesulfonamide

Example 23 Synthesis of N- (3- (1H-imidazol-1-yl) propyl) -4-methylbenzenesulfonamide

< Example  24> N- (3- (1H-Imidazol-1-yl) propyl) -3,4- Dimethylbenzenesulfonamide

The chemical structures of the compounds according to Examples 1-24 are shown in Table 1 below.

Chemical structural formula Chemical structural formula One

13

2

14

3

15

4

16

5

17

6

18

7

19

8

20

9

21

10

22

11

23

12

24

< Experimental Example  1 > AP1- luciferase  reporter, NFkB - luciferase  Activity evaluation utilized

In order to evaluate the inhibitory activity of the compound of formula (1) according to the present invention on Pin1 isomerase, the following experiment was carried out using AP1-luciferase and NFkB-luciferase reporter.

Of NFkB and AP1 transcription factor that increases the activity by Pin1 luciferase reporter vector 2.2㎍, pcDNA- Pin1 DNA 1㎍, Renilla luciferase DNA 300ng the ^{10㎕ FuGENE ® HD (Promega, WI} , USA), was placed a 1ml DMEM 20 Minute, and then 10 [mu] l of liposome-DNA mixture per well was dispensed into a 96-well plate. Here, 293T cells were dispensed at 50,000 / 100 μl per well, and cultured in a 37 ° C incubator for 24 hours. Then, the compounds of Example 1-7 were treated with cells at a concentration of 1 μM. Juglone (5-hydroxy-1,4-naphthalenedione) was used as a control inhibitor. After incubation for 48 hours in an incubator, the activity of the reporter was measured. Forty-eight hours after transplantation, 40 μl of Luciferase substrate solution was added to 40 μl of DMEM (10% serum + 1% antibiotics) medium, and left for 3 minutes. 50 μl of the reaction solution was transferred to a 96-well plate (Corning, CA, USA) and luciferase activity was measured using a microplate reader (Infinite M200 pro, TECAN, Switzerland).

Table 2 shows the results of analysis of AP1 transcription factor activity, and Table 3 shows the results of NF-kB transcription factor activity analysis.

Example activation (%) Control 100.0 Juglone 62.4 One 66.3 2 45.8 3 51.9 4 51.9 5 66.1 6 58.7 7 65.7 8 55.9 9 64.5 10 56.5 11 64.8 12 52.6 13 49.0 14 57.2 15 66.3 16 80.3 17 74.7 18 76.2 19 68.1 20 51.2 21 58.8 22 56.2 23 50.9 24 56.6

Example activation (%) Control 100.0 Juglone 51.6 One 54.6 2 29.6 3 55.7 4 65.9 5 71.5 6 21.5 7 27.1 8 31.1 9 26.1 10 22.0 11 26.8 12 33.6 13 36.9 14 59.3 15 66.8 16 34.2 17 38.4 18 33.7 19 36.8 20 39.1 21 35.9 22 28.7 23 27.8 24 49.0

As shown in Table 2, the compounds of Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 19, 20, 21 , 22, 23, and 24 were found to be superior in inhibiting the activity of AP1 transcription factor activated by Pin1 isomerase. Particularly, Examples 2, 3, 4, 6, 8, 10, 12, 13, 14, 20, 21, 22, 23 and 24 showed a better effect than the positive control group Juglone Induced a decrease in AP1 activity by more than 50% at a concentration of 1 [mu] M.

In the case of Table 3, the compounds of Examples 1, 2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17, 18, 19, 20, 21, 22, 23, 24 It was shown that the inhibitory effect of NF-kB transcription factor, which is activated by Pin1 isomerase, by more than 40%. Particularly, Examples 2, 6, 7, 8, 9, 10, 11, 12, 13, 16, 17, 18, 19, 20, 21, 22, 23 and 24 showed a better effect than the positive control group Juglone . In the case of 2, 6, 7, 9, 10, 11, 22, and 23, it induced more than 70% reduction of NF-kB activity at a concentration of 1 μM.

<Experimental Example 2> Evaluation of in vitro enzyme activity using Pin1 enzyme

In order to evaluate the inhibition of the activity of the compound represented by the formula (1) according to the present invention on the Pin1 isomerase, the following experiment was carried out using the Pin1 enzyme. Dissolved in purified (Pierce Thermo scientific inc) Pin1 protein and Trypsin is 35mM Hepes pH 7.5 buffer, substrate (H-Trp-Phe-Tyr -Ser (PO 3 H 2) -Pro-Arg-pNA) is 0.47M LiCl / It is dissolved in TFE (Trifluoroethanol) (Bachem L-2075). Pin1 inhibitor is diluted 3-fold and diluted to 10 different concentrations (200 μM-0.01 μM) and added to 50 μl of each well in a 96-well plate. Subsequently, add 16 μl of substrate (1 mg / ml) and add 5 μl of purified Pin1 protein (10 μM). Finally, 5 μl of Trypsin (0.1 mg / ml) is added and measured at 390 nm and 510 nm wavelength using a microplate reader. It was obtained for fitting, IC ₅₀ IC ₅₀ value is the measured value using Graphpad Prism program of each of Pin1 inhibitor, was used as Control Juglone inhibitors.

Example IC ₅₀ (uM) Juglone 2.178 One 16.21 2 31.15 6 2.733

In Table 4, the inhibitory effect of Pin1 isozinase activity on the inhibition of NF-kB transcription factor activity and the inhibition of the activity of AP1 transcription factor by selecting Example Compounds 1, 2, and 6 was examined. As a result, The compound showed an IC ₅₀ = 2.733 μM similar effect to the positive control group Juglone.

Accordingly, the compound represented by the formula (I) according to the present invention, an optical isomer thereof or a pharmaceutically acceptable salt thereof is excellent in the effect of inhibiting the activity of Pin1 isomerase, and therefore, And may be usefully used as an emulsion composition.

< Experimental Example  3> Evaluation of cell proliferation inhibitory activity and protein expression level of cervical cancer and pancreatic cancer cell line

The following experiments were carried out to evaluate the inhibitory activity of the compound represented by the formula (1) according to the present invention on normal Pin1 expression or knock-down cervical cancer and pancreatic cancer cell lines.

HeLa cells (normal expression of Pin1 (shCon) or low expression (shPin1), manufacturer of ATCC) and pancreatic cancer cell line were added to a 96-well plate at a concentration of 10,000 cells / well and cultured for 24 hours, Lt; / RTI &gt; The compounds of Example 1 were dissolved in the medium at concentrations of 0.032 μM, 0.16 μM, 0.8 μM, 4 μM and 20 μM, respectively. The cells were then replaced with well plates containing cells and further incubated for 72 hours in an incubator. Promega, WI, USA). The results are shown in FIG. 1 (a).

In addition, the expression levels of proteins were analyzed by using Pin1, PKM2, Cyclin D1, and GAPDH antibodies before and after Pin1 normal or knock-down cervical cancer and pancreatic cancer cell lines.

Fig. 1 (a) is a graph showing the inhibition rate of cell growth after HeLa cells treated with the compound of Example 1 at different concentrations.

Fig. 1 (b) shows the expression of Pin1 protein in HeLa cells (normal Pin1 expression (shCon) or low expression (shPin1)).

1 (C) is a graph showing the apoptosis of HeLa cells (normal Pin1 expression (shCon) or low expression (shPin1)) treated with the compound of Example 1 at different concentrations.

As shown in Fig. 1, the compound of Example 1 according to the present invention showed a concentration-dependent inhibition of (shCon) cell proliferation of HeLa, a normal cervical cancer cell line with Pin1. However, in the cell line with low Pin1 expression (shPin1), it was not significantly affected by cell proliferation even when the concentration of the drug increased. These results show that the drug acts selectively only in a normal cervical cancer cell line with Pin1 (HeLa shCon).

FIG. 2 is a graph showing Pin1 protein expression level in pancreatic cancer cell line and cell growth inhibition rate after treatment of pancreatic cancer cell line according to concentration of compound of Example 1. FIG.

FIG. 3 is a graph showing the selective expression of Pin1 target protein after treating the cervical cancer cell line HeLa (normal Pin1 expression (shCon) or low expression (shPin1)) according to the concentration of the compound of Example 1.

As shown in Fig. 2, the expression of Pin1 protein was observed in pancreatic cancer cell line. As a result, Pin1 expression was found to be high in six of 8 cell lines including Capan1, MIA Paca2, Panc1, HPAC, SU86.86 and BXPC3 , And the compound of Example 1 according to the present invention was found to inhibit cell proliferation in a concentration-dependent manner in most pancreatic cancer cell lines, except for Capan2 cell line with low expression of Pin1. Compared with the control group, Juglone, the inhibitory effect was superior to that of ATRA in most cell lines such as Mia Paca2, Panc1, BxPC3, ASPC1 and SU86.86, except for some cells.

FIG. 3 is a graph showing the selective expression of the Pin1 target protein after treating the compound of Example 1 with the cervical cancer cell line HeLa (normal Pin1 expression (shCon) or low expression (shPin1)) at different concentrations. (PKM2, CyclinD1) expression of the Pin1 target protein was decreased.

<Experimental Example 4> Confirmation of compound and Pin1 protein binding

In order to confirm the physical binding between the compound represented by the formula (1) according to the present invention and the Pin1 protein and analyze their binding ability, the following experiment was carried out using Biacore T200.

Specifically, the Pin1 recombinant protein with GST was expressed using E. coli, and the protein was purified through a GST column. The Pin1 protein was attached to the S-series CM5 chip using the amine coupling technique. The compound of Example 6 was diluted in a mobile phase buffer to prepare a mobile phase buffer. The mobile phase buffer used was 2% DMSO, 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA and 0.05% Was run at 25 [deg.] C at 30 [mu] l / min. The signal changes due to the binding of the compound of Example 6 with the Pin1 protein were confirmed by concentration as shown in FIG. 4, and the approximate binding force was analyzed using the Biacore T200 evaluation software.

FIG. 4 is a graph showing the binding of Pin1 protein to the compound of Example 6 treated at different concentrations. As a result, it was confirmed that the binding to the protein was increased by increasing the concentration of the compound.

&Lt; Experimental Example 5 > Measurement of the ability of compounds to inhibit the migration of pancreatic cancer cell lines

The following experiments were conducted to evaluate the ability of the compounds of formula (I) according to the present invention to inhibit the migration of pancreatic cancer cell lines.

Specifically, the pancreatic cancer cell line BxPC3 (manufacturer ATCC) cell line was put into a 96-well image-lock plate (essenbioscience) at a concentration of 70,000 cells / well and cultured for 24 hours to attach the cells to the plate. The compound of Example 6 and the control ATRA compound were dissolved at a concentration of 10 μM in the above medium. The wells containing the cells were wounded using the scratch wound assay kit, and the diluted compound medium Was added. Thereafter, the plate was mounted on Incucyte equipment inside the cell incubator, and the cells were further cultured every hour while images were taken. Cell migration was measured using Incucyte's analysis program, and the result after 12 hours was shown in FIG.

FIG. 5 is a graph showing the cell migration after treating the compound of Example 6 with the ATRA compound as a control group and BxPC3 as a pancreatic cancer cell line. FIG.

As shown in FIG. 5, in the pancreatic cancer cell line BxPC3, the migration ability of the cell line according to the treatment of the compound of Example 6 was evaluated. As a result, it was found that after 12 hours of the compound treatment, the DMSO and ATRA- In contrast, the compound-treated group of Example 6 showed a marked inhibition of the migration ability of BxPC3.

Accordingly, the compound represented by formula (I) according to the present invention, its optical isomer or its pharmaceutically acceptable salt has excellent effect of inhibiting the cell proliferation of cervical cancer and pancreatic cancer cell as well as activity of Pin1 isomerase May be useful as a pharmaceutical composition for preventing or treating cervical cancer or pancreatic cancer.

Meanwhile, the compound represented by Formula 1 according to the present invention can be formulated into various forms according to the purpose. Hereinafter, some formulation methods in which the compound of Formula 1 according to the present invention is contained as an active ingredient are exemplified, and the present invention is not limited thereto.

&Lt; Formulation Example 1 > Preparation of pharmaceutical preparation

1-1. Manufacture of Powder

500 mg of the compound of formula (1)

Lactose 100 mg

10 mg of talc

The above components are mixed and filled in airtight bags to prepare powders.

1-2. Manufacture of tablets

500 mg of the compound of formula (1)

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

1-3. Manufacture of capsules

500 mg of the compound of formula (1)

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

1-4. Injection preparation

500 mg of the compound of formula (1)

Sterile sterilized water for injection

pH adjuster

(2 ml) per ampoule in accordance with the usual injection method.

1-5. Manufacture of liquid agent

100 mg of the compound of formula (1)

10 g per isomer

5 g mannitol

Purified water quantity

Each component was added to purified water in accordance with the usual liquid preparation method and dissolved, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 ml with purified water, The liquid is prepared by sterilization.

Claims (7)

A pharmaceutical composition for preventing or treating a cancer or chronic inflammatory disease comprising a compound represented by the following formula (1), an optical isomer thereof or a pharmaceutically acceptable salt thereof as an active ingredient:
[Chemical Formula 1]

(In the formula 1,
R ¹ is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, or C _1-10 straight or branched alkylaminocarbonyl;
R ² is hydrogen, halogen, C _1-10 linear or branched alkyl, C _1-10 straight or branched alkoxy, C _1-10 straight or branched chain haloalkoxy, or C _1-10 straight or branched alkylthio ;
R ³ and R ⁴ are independently hydrogen, halogen, C _1-10 straight or branched chain alkyl, or C _1-10 straight or branched alkoxy;
Wherein R ¹ and R ² together with the carbon atoms to which they are attached may form a 5-7 membered heteroaryl comprising at least one heteroatom selected from the group consisting of N, O and S, Aryl may be substituted with at least one member selected from the group consisting of C _1-10 linear or branched alkyl, oxo, and C _1-10 straight chain or branched alkylcarbonyl;
Wherein R &lt; ³ &gt; and R &lt; ⁴ &gt; together with the carbon atoms to which they are attached may form a C _6-12 aryl.
The method according to claim 1,
R ¹ is hydrogen, halogen, C _1-4 straight or branched alkyl, C _1-4 straight or branched alkoxy, or C _1-4 straight or branched alkylaminocarbonyl;
R ² is hydrogen, halogen, C _1-4 straight or branched chain alkyl, C _1-4 straight or branched alkoxy, C _1-4 straight or branched chain haloalkoxy, or C _1-4 straight or branched alkylthio ;
R ³ and R ⁴ are independently hydrogen, halogen, C _1-4 straight or branched chain alkyl, or C _1-4 straight or branched alkoxy;
Wherein R ¹ and R ² together with the carbon atoms to which they are attached may form a 5-6 membered heteroaryl comprising at least one heteroatom selected from the group consisting of N, O and S, Aryl may be substituted with at least one member selected from the group consisting of C _1-4 straight or branched chain alkyl, oxo, and straight or branched C _1-4 alkylcarbonyl;
Wherein R &lt; ³ &gt; and R &lt; ⁴ &gt; together with the carbon atoms to which they are attached can form a C _6-8 aryl.
The method according to claim 1,
R ¹ is hydrogen, F, Cl, methyl, ethyl, methoxy, ethoxy, or isopropylaminocarbonyl;
R ² is hydrogen, F, Cl, methyl, methoxy, ethoxy, trifluoromethoxy, or methylthio;
R ³ is hydrogen;
R ⁴ is hydrogen, Cl, methyl, methoxy or ethoxy, and;
Said R &lt; ¹ &gt; and R &lt; ² &gt; together with the carbon atoms to which they are attached may form a pyrrolidine substituted with morpholine or methylcarbonyl substituted with methyl and oxo groups;
Wherein R &lt; ³ &gt; and R &lt; ⁴ &gt; together with the carbon atoms to which they are attached can form a phenyl.
The method according to claim 1,
The pharmaceutical composition according to claim 1, wherein the compound represented by the formula (1) is any one selected from the group consisting of the following compounds:
1) N- (3- (1H-imidazol-1-yl) propyl) -4-chloronaphthalene-1-sulfonamide;
2) N- (3- (1H-imidazol-1-yl) propyl) -4-methoxy-3-methylbenzenesulfonamide;
3) N- (3- (1H-imidazol-1-yl) propyl) -4,5-dichloro-2-methylbenzenesulfonamide;
4) N- (3- (1H-imidazol-1-yl) propyl) -4-fluoro-3-methylbenzenesulfonamide;
5) N- (3- (1H-imidazol-1-yl) propyl) -2,5-dichlorobenzenesulfonamide;
6) N- (3- (1H-imidazol-1-yl) propyl) -4-butoxybenzenesulfonamide;
7) N- (3- (1H-imidazol-1-yl) propyl) -5-fluoro-2-methoxybenzenesulfonamide;
8) N- (3- (1H-imidazol-1-yl) propyl) -2,5-diethoxy-4-methylbenzenesulfonamide;
9) N- (3- (1H-imidazol-1-yl) propyl) -2-methoxy-4,5-dimethylbenzenesulfonamide;
10) N- (3- (1H-imidazol-1-yl) propyl) -4- (trifluoromethoxy) benzenesulfonamide;
11) N- (3- (1H-imidazol-1-yl) propyl) -4-ethoxybenzenesulfonamide;
2-methyl-3-oxo-3,4-dihydro-2H-benzo [b] [1,4] oxazine- - sulfonamide;
13) N- (3- (1H-imidazol-1-yl) propyl) -3-fluoro-4-methoxybenzenesulfonamide;
14) N- (3- (1H-imidazol-1-yl) propyl) -3-ethyl-4-methoxybenzenesulfonamide;
15) N- (3- (1H-imidazol-1-yl) propyl) -5-chloro-2-methoxybenzenesulfonamide;
16) N- (3- (1H-imidazol-1-yl) propyl) -4-methoxynaphthalene-1-sulfonamide;
17) N- (3- (1H-imidazol-1-yl) propyl) -1-acetylindoline-5-sulfonamide;
18) 5- (N- (3- (1H-Imidazol-1-yl) propyl) sulfamoyl) -N-isopropyl-2-methylbenzamide;
19) N- (3- (1H-imidazol-1-yl) propyl) -4- (methylthio) benzenesulfonamide;
20) N- (3- (1H-imidazol-1-yl) propyl) -3-chloro-4-methoxybenzenesulfonamide;
21) N- (3- (1H-imidazol-1-yl) propyl) -3-ethoxy-4-fluorobenzenesulfonamide;
22) N- (3- (1H-imidazol-1-yl) propyl) -4-chloro-3-methoxybenzenesulfonamide;
23) N- (3- (1H-imidazol-1-yl) propyl) -4-methylbenzenesulfonamide; And
24) N- (3- (1H-Imidazol-1-yl) propyl) -3,4-dimethylbenzenesulfonamide.
The method according to claim 1,
Wherein said cancer is one selected from the group consisting of cervical cancer, colon cancer, pancreatic cancer, ovarian cancer, and cervical cancer.
The method according to claim 1,
Wherein the chronic inflammatory disease is asthma.
The method according to claim 1,
Wherein said pharmaceutical composition inhibits the Pin1 isomerization enzyme.

Images