KR20210108684A - Novel Pim Kinase Inhibitor and Uses Thereof - Google Patents

Abstract

The present invention relates to a novel Pim kinase inhibitor and use thereof. Particularly, the present invention relates to a novel compound based on 1,3,4-oxadiazol-2(3H)-thione having an activity of inhibiting Pim kinase, and a composition for inhibiting Pim kinase or composition for preventing or treating cancer, including the compound. The compound based on 1,3,4-oxadiazol-2(3H)-thione according to the present invention is shown to have an activity of inhibiting Pim kinase highly effectively even at a low concentration. Therefore, the compound according to the present invention can be used advantageously for a composition for preventing or treating cancer by inhibiting Pim kinase.

Description

Novel Pim Kinase Inhibitor and Uses Thereof

The present invention relates to a novel Pim kinase inhibitor and uses thereof, and more particularly, to a novel 1,3,4-oxidazole-2(3H)-thione (1,3,4-Oxadiazole having Pim kinase inhibitory activity). -2(3H)-thione)-based compound and a composition for inhibiting Pim kinase comprising the same, or a composition for preventing or treating cancer.

Pim kinases regulate cellular signaling pathways involved in cancer development and progression (Amaravadi and Thompson, 2005). They are members of the serine/threonine kinase family, which consists of three isoforms, Pim-1, Pim-2 and Pim-3. Pim kinase is overexpressed in hematologic cancers and is known to phosphorylate downstream that contributes to tumor growth and survival. Inhibitors of these kinases are expected to treat cancers associated with high expression of Pim kinase, and several Pim kinase inhibitors are currently undergoing clinical trials (Anizon, F., et al. , Curr . Med . Chem ., 17 : 4114-33, 2010; Lee, J., et al. , Chem . Pharm . Bull., 62 : 906-914, 2014; Morwick, T. Expert Opin. Ther. Pat., 20:193-212, 2010) .

According to the crystal structure of Pim-1 and Pim-2 kinases, the Pim kinase family has a unique proline (Pro123 of Pim-1) residue in the hinge region of the enzyme, and is associated with ATP compared to other serine/threonine kinases. One hydrogen bond donor for hydrogen bond interaction is lacking (Kumar, A., et al. , J. Mol . Biol . , 348:183-193, 2005). Therefore, Pim kinase can make only one hydrogen bond to adenine of ATP, which is a unique property of Pim kinase, which can be used to discover inhibitors with high selectivity compared to other kinases.

Most of the reported Pim kinase inhibitors were designed to be selective inhibitors for Pim kinase by using the interaction with the amino residue of lysine (Lis67 of Pim-1) at the Pim kinase ATP binding site as an alternative driving force for binding (Bullock, AN). , et al. , J. Biol . Chem . , 280:41675-41682, 2005), the inventors of the present invention, in the process of developing a selective Pim kinase, 1,3,4-oxidazole-2(3H)-thione ( 1,3,4-Oxadiazole-2(3H)-thione) was identified as a novel inhibitor interacting with the α-amino residue of lysine (Lee, AY, et al ., QBS, 37:113-124, 2018) ).

Therefore, in the present invention, as a result of intensive efforts to develop a more effective Pim kinase inhibitor, 1,3,4-oxidazole-2(3H)-thione (1,3,4-Oxadiazole-2(3H) at the 3rd position of the indole )-thione) was synthesized as a new Pim kinase inhibitor introducing an alternative substance to the 6th position substituent of the introduced compound, and it was confirmed that these compounds had effective Pim kinase inhibitory activity even at low concentrations, and completed the present invention.

It is an object of the present invention to provide a 1,3,4-oxidazole-2(3H)-thione-based compound having Pim kinase inhibitory activity and a method for preparing the same.

Another object of the present invention is to provide a composition for inhibiting Pim kinase comprising the compound.

Another object of the present invention is to provide a composition for preventing or treating cancer comprising the compound.

In order to achieve the above purpose,

The present invention provides a compound represented by the following formula (1) having a Pim kinase inhibitory activity.

[Formula 1]

And in the formula 1 R is a halogen, R ₁ an optionally substituted aromatic or heteroaromatic group, wherein R ₁ is an alkyl amine, a cyclic amine or an alcohol side-substituted with halogen or R _2, wherein R ₂ is an alkyl amine group am.

In a preferred embodiment of the present invention, the compound may be represented by any one selected from the group consisting of the following Chemical Formulas 2 to 10.

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

In another preferred embodiment of the present invention, the compound may be specifically bound to the ATP binding site of Pim kinase.

In another preferred embodiment of the present invention, the Pim kinase may be Pim-1, Pim-2 or Pim-3.

In order to achieve another object, the present invention provides a Pim kinase inhibitory composition comprising the compound having the Pim kinase inhibitory activity.

In order to achieve another object, the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the compound having the Pim kinase inhibitory activity.

In addition, the present invention provides a pharmaceutical composition for the treatment of diseases or disorders mediated by Pim kinase, comprising the compound having the Pim kinase inhibitory activity.

According to a preferred embodiment of the present invention, the disease or disorder mediated by the Pim kinase may be selected from immune disorders, cardiovascular diseases, viral infections, inflammation, metabolic/endocrine dysfunction and neurological disorders.

It was confirmed that the 1,3,4-oxidazole-2(3H)-thione (1,3,4-Oxadiazole-2(3H)-thione)-based compound of the present invention has a very effective Pim kinase inhibitory activity even at a low concentration Therefore, the compound of the present invention can be usefully used as a composition for preventing or treating cancer through inhibition of Pim kinase.

1 is a schematic diagram showing the docking mode of the compound 3 of the present invention to the Pim-1 (PDB 4DTK) kinase ATP binding site. In compound 3, 4 hydrogen bonds between the Pim-1 residues (Lys67, Glu121, Aps128 and Glu171) are expected, which are indicated by green dotted lines.
2 is a schematic diagram showing Scheme 1 for synthesizing the compound of the present invention.
3 is a schematic diagram showing Scheme 2 for synthesizing the compound of the present invention.
4 is a schematic diagram showing Scheme 3 for synthesizing the compound of the present invention.
5 is a schematic diagram showing Scheme 4 for synthesizing the compound of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention, from one point of view, relates to a compound represented by the following formula (1) having a Pim kinase inhibitory activity.

[Formula 1]

And in the formula 1 R is a halogen, R ₁ an optionally substituted aromatic or heteroaromatic group, wherein R ₁ is an alkyl amine, a cyclic amine or an alcohol side-substituted with halogen or R _2, wherein R ₂ is an alkyl amine group am.

Preferably, the compound may be represented by any one selected from the group consisting of the following Chemical Formulas 2 to 10.

[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

In the present invention, the compound may be characterized in that it specifically binds to the ATP binding site of Pim kinase.

In the present invention, the Pim kinase may be characterized as Pim-1, Pim-2 or Pim-3.

The heteroaromatic ring, 1,3,4-oxadiazole-2(3H)thione, has an acidic proton with a pKa value of 6.5 (ChemAxon) when it has a phenyl group at the 5-position, and is expected to be deprotonated under physiological conditions. As expected, the deprotonated form interacts with the e-aminoganji of lysine in the ATP binding pocket and may cause its derivatives to inhibit the catalytic activity of Pim kinase.

In the present invention, in order to develop a Pim kinase inhibitor, a derivative having 1,3,4-oxadiazole-2(3H)thione at the 3-position of the indole was prepared in Scheme 1 (FIG. 2), Scheme 2 (FIG. 3) and It was synthesized as in Scheme 3 (FIG. 4).

First, as shown in Scheme 1, compound 2 was synthesized from 5-bromo-1H-indole, and compound A was acylated with 2,2,2-trichloroacetyl chloride in methanol, and then the ester was converted to hydrazide. _{After cyclization with CS 2} under basic conditions, compound 2 was obtained.

Compounds 3 to 10 were synthesized as shown in Scheme 2 by replacing the bromine atom with arene.

Specifically, the tosyl group of Compound A was protected, and palladium-catalyzed borylation was performed with bis(pinacolato)diboron. Compound E was obtained by Suzuki coupling reaction between compound D and aryl halide, which was deprotected with a base to obtain compound F. After converting the ester to hydrazide, compounds 3 to 10 were synthesized by cyclization with _{CS 2 under basic conditions.}

In particular, compounds 3 to 8 are pyrazine and pyridine having an aminoalkyl substituent at the 5-position of the indole ring attached to the 5-position of the indole ring, Compound H-1 Substituents of compounds H-6 were synthesized by a coupling reaction of 2,6-dichloropyrene or 2,6-dithloropyridine with the corresponding amine or alcohol according to Scheme 4.

In addition, in the present invention, S-alkylated compounds Compound 11 and Compound 12 were synthesized from Compound 9 or Compound 10 as shown in Scheme 4, and benzyl bromide was added under basic conditions to form an S-benzyl compound. 11 and compound 12 were synthesized.

In a specific embodiment of the present invention, compounds 2 to 10, compounds 11 and 12 were synthesized as shown in Schemes 1 to 3, and Pim kinase inhibitory activity was measured for them.

As shown in Table 1, compound 2, which is a prototype of an indole and 1,3,4-oxadiazole-2(3H)-thione conjugate, has inhibitory activity at submicromolar concentrations against all three Pim kinases. , and it was confirmed that it showed a significant inhibitory effect on Pim-1 and Pim-3 rather than Pim-2.

Compound 9 and compound 10, which are phenyl or 2-fluorophenyl substituted compounds, showed reduced Pim kinase inhibitory activity compared to compound 2, indicating that a bulky hydrophobic group at the 5-position of the indole ring is not suitable. means that

In addition, it was confirmed that compounds 11 and 12 also had reduced Pim kinase inhibitory activity compared to other compounds.

In the case of compounds 3 to 8 in which pyrazine and pyridine having an aminoalkyl substituent are attached to the 5-position of the indole ring, Pim-1 and Pim- by introduction of a heteroaromatic ring It was found to be a very potent Pim kinase inhibitor with _{very low nanomolar IC 50 values for 3 kinases (Table 2).}

The type of linker between the aminoalkyl group and the pyrazine also affected the inhibitory activity of Pim kinase, and the amino linkage had twice the inhibitory activity on Pim-1 and Pim-3 kinases compared to the ether linkage. appeared to be

The compound in which the 5-position of the indole ring is substituted with phenyl induced a decrease in Pim kinase inhibitory activity, while both pyrazine and pyridine increased the Pim kinase inhibitory activity, indicating that pyrazine and pyridine are more likely to bind to Pim kinase. It means structure.

In another specific embodiment of the present invention, in order to analyze the molecular docking of compound 3 and Pim-1 (PDB 4DTK) kinase ATP binding site, the crystal structure of PCS-1 kinase (PDB ID: 4DTK) was analyzed from RCSB protein data. obtained from the bank. As shown in FIG. 1 , as a result of performing molecular docking on compound 3, it was confirmed that four hydrogen bond interactions and some hydrophobic interactions may exist between compound 3 and the Pim-1 protein.

In another specific embodiment of the present invention, the selectivity for the kinase of compound 4 was confirmed using the ^{Kinase Profiler TM project of Eurofins.} As shown in Table 3, compound 4 showed a weak activity inhibitory effect on Flt3, GSK3β and JNK3 kinase, but _{considering the very low IC 50} value of compound 4 for Pim-1 and Pim-3, compound 4 was It can be seen that it has very high selectivity for Pim-1 and Pim-3.

That is, the 1,3,4-oxidazole-2(3H)-thione (1,3,4-Oxadiazole-2(3H)-thione)-based compound of the present invention can be used as a Pim kinase activity inhibitor, and the Pim kinase Since modulates cellular signals related to cancer development and progression, the compound of the present invention can be usefully used as a composition for preventing or treating cancer by inhibiting Pim kinase.

Accordingly, the present invention relates to a Pim kinase inhibitory composition comprising the compound having the Pim kinase inhibitory activity from another aspect.

In another aspect, the present invention relates to a pharmaceutical composition for preventing or treating cancer comprising the compound having the Pim kinase inhibitory activity.

The composition may further include other anticancer agents or kinase inhibitors for more effective cancer prevention or treatment, and may be administered in parallel with other anticancer therapies.

The cancer is multiple myeloma, breast cancer, ovarian cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, skin cancer, keratoacanthoma, lung cancer, epidermal carcinoma, large cell carcinoma , non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, adenocarcinoma, thyroid cancer, follicular carcinoma, undifferentiated carcinoma, papillary carcinoma, seminoma, malignant melanoma, sarcoma, bladder carcinoma, liver and biliary tract carcinoma, renal carcinoma, pancreatic cancer, bone marrow disorder, lymphoma, hair cell cancer, buccal cancer, nasopharyngeal cancer, pharyngeal cancer, labial cancer, tongue cancer, oral cancer, small intestine cancer, colon-rectal cancer, colorectal cancer, rectal cancer, brain cancer and central nervous system cancer Tumor, Hodgkin's disease, hematologic cancer, leukemia, bronchial cancer, thyroid cancer, liver and bile duct cancer, hepatocellular carcinoma, gastric cancer, glioma/glioblastoma, endometrial cancer, malignant melanoma, kidney and renal pelvic cancer, bladder cancer, uterine fibroids, can be selected from the group consisting of cervical cancer, acute myeloid leukemia, chronic myelogenous leukemia, lymphocytic leukemia, myeloid leukemia, oral and pharyngeal cancer, non-Hodgkin's lymphoma, malignant melanoma, and chorionic colon adenoma, but inhibiting Pim kinase activity It can be used without limitation for cancers that have anticancer effects.

In another aspect, the present invention relates to a pharmaceutical composition for treating a disease or disorder mediated by Pim kinase.

The disease or disorder mediated by the Pim kinase may be selected from immune disorders, cardiovascular diseases, viral infections, inflammation, metabolic/endocrine dysfunction and neurological disorders.

The pharmaceutical composition of the present invention may be formulated in various forms according to a conventional method and used. For example, it may be formulated in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, and may be formulated in the form of external preparations, suppositories, and sterile injection solutions. It may further include a pharmaceutically acceptable carrier, excipient and diluent depending on each formulation. In addition, according to a conventional method, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc. and sterile injection solutions for external use.

The carrier, excipient and diluent include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, Microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, mineral oil, and the like. When formulating or formulating the pharmaceutical composition, it is usually prepared using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the composition, for example, starch, calcium carbonate, sucrose. , lactose, gelatin, etc. are mixed and prepared. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid preparations for oral use include suspensions, solutions, emulsions, syrups, etc., and various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included in addition to commonly used simple diluents such as water and liquid paraffin. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

As used herein, the term 'administration' means providing the pharmaceutical composition of the present invention to an individual by any suitable method. The pharmaceutical composition of the present invention provides an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human as thought by a researcher, veterinarian, doctor or other clinician, that is, alleviation of symptoms of a disease or disorder to be treated. It can be administered in a therapeutically effective amount, which is an amount that induces It is apparent to those skilled in the art that the therapeutically effective dosage and frequency of administration for the pharmaceutical composition of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, the age, weight, and general health of the patient , sex and diet, administration time, administration route and secretion rate of the composition, treatment period, and various factors including concurrently used drugs. The pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg/kg/day, may be administered once a day, or may be administered in divided doses.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

Synthesis of novel compounds having Pim kinase inhibitory activity according to Scheme 1

In the present invention, 1,3,4-oxidazole-2(3H)-thione-based compounds were synthesized, and 1H-NMR and 13C-NMR spectra of the synthesized compounds were obtained from Bruker Spectrospin 400 (400 MHz) or JEOL ECA 500 (500 MHz). Analyzes were performed using a spectrometer. Chemical shifts (δ was analyzed in ppm using tetramethylsilane as an internal standard. Mass spectra were analyzed using Waters ACQUITY UPLC and Micromass Quattro micro ^TM AP.

Microwave assisted reaction was performed using CEM Discover BenchMate. TLC was performed using E. Merck silica gel 60 F254 plates (0.25 mm), and silica gel column chromatography was performed using Merck silica gel 60 (230-400 mesh).

Unless otherwise stated, all starting materials were purchased from commercially available sources and used without further purification. In addition, all reactions were carried out under nitrogen conditions.

<Synthesis of Compound 2>

Compound A: methyl -5- Bromo -1H-Indole-3- carboxylate (Methyl 5- bromo -1H-indole-3-carboxylate)

According to Scheme 1, first 5-bromoindole (5-Bromoindole; 5.1 mmol, 1.0 g), 4-dimethylaminopyridine (4-dimethylaminopyridine; DMAP; 0.51 mmol, 0.062 g) and pyridine (pyridine; 17 mmol, 1.4 mL) ) was mixed, and then tetrahydrofuran (THF, 5.0 mL) dried at 0° C. was added. 2,2,2-trichloroacetyl chloride (2,2,2-trichloroacetyl chloride; 15 mmol, 1.7 mL) was slowly added to the mixture at 0°C. After stirring at room temperature overnight, 5.0 mL of methanol was slowly added to the reaction mixture at 0°C. After potassium carbonate (K ₂ CO ₃ ; 3.0 mmol, 4.2 g) was added, the mixture was refluxed for 5 hours. After removal of the solvent in vacuo, the residue was treated with 1M hydrochloric acid solution until pH 2 was reached.

The product was extracted with ethyl acetate (EA), and the extract was washed with water and brine. The organic layer was dried over anhydrous magnesium sulfate (MgSO ₄ ), and the solvent was removed in vacuo.

The residue was purified by silica gel chromatography using 1:4 EA/Hexane (n-Hex) and then repurified by silica gel chromatography using 1:1 EA/n-Hex to obtain 0.85 g of compound A (66 %) was obtained.

The analysis results of compound A are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.14 (s, 1H), 8.15 (s, 1H), 8.12 (d, J = 1.7 Hz, 1H), 7.47 (d, J = 8.6 Hz) , 1H), 7.34 (dd, J = 8.6, 2.3 Hz, 1H), 3.82 (s, 3H).

Compound B: 5- Bromo -1H-Indole-3- Carbohydrazine (5- Bromo -1H- indole -3-carbohydrazide)

A microwave vessel was charged with the compound A (1.2 mmol, 0.30 g) and ethanol (EtOH; 1.0 mL), hydrazine hydrate (12 mmol, 0.37 mL) was added, and then 100 W was applied to the reaction mixture to 120 Irradiated for 10 minutes at °C.

The solvent was removed in vacuo, and the residue was purified by silica gel chromatography using _{1:10:100 ammonium hydroxide (NH 4} OH)/methanol (MeOH)/chloroform (CHCl _{3 ).} , 1:10:80 NH ₄ OH/MeOH/CHCl ₃ was repurified by silica gel chromatography to obtain 0.23 g (76 %) of Compound B.

The analysis results of compound B are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.73 (s, 1H), 9.26 (s, 1H), 8.29 (d, J = 1.1 Hz, 1H), 8.01 (d, J = 2.9 Hz) , 1H), 7.41 (d, J = 8.6 Hz, 1H), 7.27 (dd, J = 8.6, 1.7 Hz, 1H), 4.35 (s, 2H).

compound 2:5 -(5- Bromo -1H-indole-3 yl)-3H-1,3,4- oxadiazole -2- thione (5-(5-Bromo-1H-indol-3-yl)-3H-1,3,4-oxadiazole-2-thion)

_{Potassium hydroxide (KOH; 0.91 mmol, 0.051 g) and carbon disulfide (CS 2} ; 1.4 mmol, 0.082 mL) were added to a solution of EtOH (5.0 mL) containing the compound B (0.91 mmol, 0.23 g) at 0 °C. was added and the reaction mixture was stirred at 0° C. for 30 min. After further stirring at room temperature for 30 min, the mixture was refluxed for 2 h. The solvent was removed in vacuo and the residue was treated with 2M HCl solution until the solution was pH 2.

The product was extracted with ethyl acetate, and the extract was washed with water and brine. The organic layer was dried over anhydrous magnesium sulfate and the solvent was removed in vacuo. Residue 1: 10: 100 NH ₄ OH / MeOH / CHCl ₃ was purified by silica gel chromatography to obtain 0.16 g (61%) of Compound 2 represented by the following Chemical Formula 2.

[Formula 2]

The analysis results of compound 2 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.57 (s, 1H), 8.04 (d, J = 1.7 Hz, 1H), 7.73 (d, J = 1.7 Hz, 1H), 7.27 (dt) , J = 8.6, 2.0 Hz, 1H), 7.21 (dt, J = 8.6, 2.0 Hz, 1H); ESI MS: m/z = 296 [M+H] ⁺ .

according to Scheme 2 Pim Synthesis of novel compounds with kinase inhibitory activity

2-1: Substituent Synthesis

Compounds 3 to 10 were prepared according to Scheme 2, and first, substituents of compounds H-1 to H-6 prepared according to Scheme 4 were synthesized for the synthesis of compounds 3 to 8.

Compound H-1: N -(6- chloropyrazine -2 days)- N', N' -Dimethyl-ethane-1,2- diamine ( N -(6-Chloropyrazin-2-yl)- N', N' -dimethyl-ethane-1,2-diamine)

N, N - dimethyl ethylene diamine was added (N, N -dimethylethylenediamine, 1.7 mmol , 0.25 g) and _{K 2 CO 3 (3.3 mmol,} 0.46 g) in 5.0 mL DMF, and the mixture was stirred at room temperature for 30 minutes . To the reaction mixture was added 2,6-dichloropyrazine (2,6-dichloropyrazine, 2.0 mmol, 0.25 g) and stirred overnight, then the solvent was removed in vacuo. The residue was treated with DCM and filtered through Celite. The solvent of the obtained filtrate was removed in vacuo, and the residue was purified by silica gel chromatography using 1:9 MeOH/DCM to obtain 0.27 g (75%) of compound H-1 represented by the following formula H-1. .

[Formula H-1]

The analysis results of compound H-1 are as follows:

¹ H-NMR (500 MHz, CDCl ₃ ): δ7.77 (s, 1H), 7.75 (s, 1H), 5.53 (s, 1H), 3.39 (q, J = 5.5 Hz, 2H), 2.55 (t) , J = 5.7 Hz, 2H), 2.26 (s, 6H).

Compound H-2: N -(6- chloropyrazine -2 days)- N', N' - diethyl -ethane-1,2- diamine ( N -(6-Chloropyrazin-2-yl)- N', N' -diethyl-ethane-1,2-diamine)

According to the synthesis process of the compound compound H-1, it was synthesized using N,N -diethylethylenediamine ( N,N- diethylethylenediamine). The product was purified by silica gel chromatography using 5:95 MeOH/DCM and 1:9 MeOH/DCM to obtain compound H-2 represented by the following formula H-2 in a yield of 87%.

[Compound H-2]

The analysis results of compound H-2 are as follows:

¹ H NMR (500 MHz, CDCl ₃ ): δ7.74 (d, J = 2.9 Hz, 2H), 3.36-3.30 (m, 2H), 2.66 (t, J = 6.3 Hz, 2H), 2.55 (q, J = 7.3 Hz, 4H), 1.01 (t, J = 7.2 Hz, 6H).

compound H- 3: 2 -(6- chloropyrazine -2 days) oxy - N, N - dimethylethanamine (2-(6-Chloropyrazin-2-yl)oxy- N, N -dimethylethanamine)

2,6-dichloropyrazine (2,6-dichloropyrazine, 2.0 mmol, 0.30 g), 2-(dimethylamino)ethanol, 2.2 mmol, 0.22 mL) and THF (2.0 mL) in a microwave vessel was added and stirred at room temperature for 10 minutes. NaH (4.0 mmol, 0.097 g) was further added to the reaction mixture, and then 100 W was applied to the mixture to irradiate the mixture at 50° C. for 10 minutes. After removal of the solvent in vacuo, the residue was treated with EA and the EA solution was washed with water and brine. The organic layer was dried over anhydrous MgSO _{4 ,} and then the solvent was removed in vacuo to obtain 0.35 g (87%) of compound H-3 represented by the following formula H-3.

[Formula H-3]

The analysis results of compound H-3 are as follows:

¹ H NMR (400 MHz, CDCl ₃ ): δ8.19 (s, 1H), 8.14 (s, 1H), 4.44 (t, J = 5.4 Hz, 2H), 2.74 (t, J = 5.4 Hz, 2H) , 2.34 (s, 6H).

compound H- 4:3 -(6- chloropyrazine -2 days) oxy - N, N -Dimethylpropane- 1 amine (3-(6-Chloropyrazin-2-yl)oxy- N, N -dimethylpropan-1-amine)

According to the above compound compound H-3 synthesis procedure, using 3-dimethylamino-1-propanol (3-dimethylamino-1-propanol), compound H-4 represented by the following formula H-4 was synthesized in 79% yield.

[Formula H-4]

The analysis results of compound H-4 are as follows:

¹ H NMR (400 MHz, CDCl ₃ ): δ8.13 (s, 1H), 8.12 (s, 1H), 4.39 (t, J = 6.4 Hz, 2H), 2.44 (t, J = 7.4 Hz, 2H) , 2.26 (s, 6H), 1.97 (sext, J = 5.5, 2H).

compound H- 5:2 - Chloro -6- (4-methylpiperazin-1-yl)pyrazine (2- Chloro -6-(4-methylpiperazin-1-yl)pyrazine)

According to the compound H-3 synthesis process, 1-methylpiperazine was used to synthesize compound H-5 represented by the following formula H-5 in a yield of 91%.

[Formula H-5]

The analysis results of compound H-5 are as follows:

¹ H NMR (400 MHz, CDCl ₃ ): δ7.98 (s, 1H), 7.80 (s, 1H), 3.63 (t, J = 5.1 Hz, 4H), 2.51 (t, J = 5.1 Hz, 4H) , 2.35 (s, 3H).

Compound H-6: N-(6- Chloro -2- pyridyl )- N', N' -Dimethylethane-1,2- diamine (N-(6-Chloro-2-pyridyl)- N', N' -dimethylethane-1,2-diamine)

According to the compound H-3 synthesis process, 2,6-dichloropyridine was used to synthesize compound H-6 represented by the following formula H-6 in 74% yield.

[Formula H-6]

The analysis results of compound H-6 are as follows:

¹ H NMR (500 MHz, CDCl ₃ ): δ7.31 (t, J = 7.7 Hz, 1H), 6.53 (d, J = 7.4 Hz, 1H), 6.26 (d, J = 8.0 Hz, 1H), 5.28 (s, 1H), 3.32 (dd, J = 11.7 Hz, 4.7 Hz, 2H), 2.53 (t, J = 6.0 Hz, 2H), 2.25 (s, 6H).

2-2: Synthesis of compounds 3 to 8

<Synthesis of compound 3>

Compound C: methyl -5- Bromo -One-( p- tolylsulfonyl ) indol- carboxylate (Methyl 5-bromo-1-( p -tolylsulfonyl)indole-3-carboxylate)

Sodium hydride (NaH, 2.0 mmol, 0.047 g) was added to 5.0 mL of DMF (dimethylformamide) solution containing the compound A (0.98 mmol, 0.25 g), and the reaction mixture was stirred at room temperature for 30 minutes. . 4-toluenesulfonyl chloride (TsCl, 1.1 mmol, 0.21 g) was added to the mixture at 0° C., and the reaction mixture was stirred at 0° C. for 3 hours, and then the solvent was removed in vacuo, The residue was treated with 2M HCl solution until the solution reached pH 2.

The product was extracted with ethyl acetate, and the extract was washed with water and brine. The organic layer was dried over anhydrous magnesium sulfate, and the solvent was removed in vacuo to obtain 0.37 g (92%) of compound C.

The analysis results of compound C are as follows:

¹ H-NMR (500 MHz, CDCl ₃ ): δ8.27 (d, J = 1.7 Hz, 1H), 8.25 (s, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.80 (d, J) = 8.6 Hz, 2H), 7.46 (dd, J = 8.6, 1.7 Hz, 1H), 7.28 (d, J = 8.6 Hz, 2H), 3.93 (s, 3H), 2.38 (s, 3H).

Compound D: methyl One-( p- tolylsulfonyl )-5-(4,4,5,5- tetramethyl -1,3,2- dioxabolol Lan-2-yl) indole-3-carboxylate (Methyl 1-( p - tolylsulfonyl )-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)indole-3-carboxylate)

In a microwave vessel, the above compound C (0.91 mmol, 0.37 g), 1,4-dioxane (1,4-dioxane, 2.0 mL), and EtOH (0.40 mL) were added, and then bis(pinacolato)diboron ( bis(pinacolato)diboron, 0.98 mmol, 0.25 g) and potassium acetate (KOAc, 2.7 mmol, 0.27 g) were further added, followed by _{purging with N 2} gas for 5 minutes. [1,1'-bis(diphenylphosphofino)ferrocene]dichloropalladium(II)([1,1′PdCl2(dppf), 0.027 mmol, 0.020 g) was added to the reaction mixture under N _{2 gas conditions,} 100 W was applied to the reaction mixture and irradiated at 110° C. for 10 minutes.

The solvent was removed in vacuo, and the residue was purified by dry column vacuum chromatography (DCVC) on silica gel containing DCM (dichloromethane) to obtain 0.40 g (97%) of compound D.

The analysis results of compound D are as follows:

¹ H-NMR (500 MHz, CDCl ₃ ): δ8.57 (s, 1H), 8.26 (s, 1H), 7.95 (d, J = 9.2 Hz, 1H), 7.82-7.78 (m, 4H), 7.28 (d, J = 8.0 Hz, 1H), 7.24 (d, J = 8.0 Hz, 2H), 3.94 (s, 3H), 2.35 (s, 3H), 1.34 (s, 12H).

Compound E: methyl 5-[6-[2-(dimethylamino) ethylamino ] pyrazine -2-yl]-1-( p - tolylsulfonyl ) Indole-3-carboxylate (Methyl 5-[6-[2-(dimethylamino)ethylamino]pyrazin-2-yl]-1-(p-tolylsulfonyl)indole-3-carboxylate)

In a microwave vessel, the compound D (0.66 mmol, 0.30 g), the compound H-1 (0.75 mmol, 0.15 g), 1,4-dioxane (1,4-dioxane, 2.0 mL), EtOH (0.40 mL) and 2M K ₂ CO ₃ (2.2 mmol, 1.1 mL) was added and then tetrakis(triphenylphosphine)palladium(0)(tetrakis(triphenylphosphine)palladium(0); Pd(PPh ₃ ) ₄ , 0.020 under nitrogen condition. mmol, 0.023 g), the mixture was irradiated at 110° C. for 10 minutes by applying 100 W.

The solvent was removed in vacuo and the residue was treated with a mixture of methanol and DCM (1:9) and then filtered through celite. After obtaining a filtrate and removing the solvent in vacuo, the residue was purified by two successive silica gel chromatography of 7:93 MeOH/DCM and 1:9 MeOH/DCM to give 0.17 g (52%) of compound E obtained.

The analysis results of compound E are as follows:

¹ H-NMR (500 MHz, CDCl ₃ ): δ8.70 (s, 1H), 8.31 (s, 1H), 8.25 (s, 1H), 8.06-8.00 (m, 2H), 7.86 (s, 1H) , 7.84 (d, J = 4.6 Hz, 2H), 7.30 (d, J = 8.0 Hz, 2H), 3.95 (s, 3H), 3.64 (t, J = 6.0 Hz, 2H), 2.75 (s, 2H) , 2.42 (s, 6H), 2.38 (s, 3H).

Compound F: methyl 5-[6-[2-(dimethylamino) ethylamino ] pyrazine -2-yl]-1 H -Indole-3-carboxylate (Methyl 5-[6-[2-( dimethylamino ) ethylamino ] pyrazin -2- yl ]-One H -indole-3-carboxylate)

The compound E (0.34 mmol, 0.17 g), 2-methoxyethanol (2-methoxyethanol; 2.0 mL) and K ₂ CO ₃ (0.69 mmol, 0.095 g) were added to a microwave vessel, and then 100 W was applied to the mixture to Irradiated at 70° C. for 10 minutes.

The solvent was removed in vacuo _{and purified by silica gel chromatography using 1:10:100 NH 4} OH/MeOH/CHCl ₃ to obtain 0.076 g (65 %) of compound F.

The analysis results of compound F are as follows:

¹ H-NMR (500 MHz, CDCl ₃ ): δ9.27 (s, 1H), 8.77 (s, 1H), 8.36 (s, 1H), 7.94 (d, J = 3.2 Hz, 1H), 7.92 (dd , J = 8.6, 1.7 Hz, 1H), 7.80 (s, 1H), 7.45 (d, J = 8.6 Hz, 1H), 5.27 (t, J = 4.6 Hz, 1H), 3.93 (s, 3H), 3.53 (q, J = 5.7 Hz, 2H), 2.59 (t, J = 5.7 Hz, 2H), 2.28 (s, 6H).

Compound G: 5-[6-[2-(dimethylamino)) ethylamino ] pyrazine -2-yl]- 1 H -Indole-3-carbohydrazide (5-[6-[2-(Dimethylamino)ethylamino]pyrazin-2-yl]-1H-indole-3-carbohydrazide)

The compound F (0.19 mmol, 0.076 g), EtOH (1.0 mL) and hydrazine hydrate (2.2 mmol, 0.067 mL) were added to a microwave vessel, and then 100 W was applied to the mixture and irradiated at 120° C. for 10 minutes. did..

The solvent was removed in vacuo _{and purified by silica gel chromatography using 1.5:15:80 NH 4} OH/MeOH/CHCl ₃ to give 0.066 g (87 %) of compound G.

compound 3: 5 -[5-[6-[2-(dimethylamino) ethylamino ] pyrazine -2-yl]-1 H -indol-3-yl]- 3 H -1,3,4- oxadiazole -2- thione (5-[5-[6-[2-( Dimethylamino ) ethylamino ] pyrazin -2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

_{Potassium hydroxide (KOH; 0.20 mmol, 0.011 g) and carbon disulfide (CS 2} , 0.41 mmol, 0.025 mL) were added to a solution of EtOH (5.0 mL) containing the compound G (0.17 mmol, 0.066 g) at 0 ° C. After addition, the reaction mixture was stirred at 0 °C for 30 min. The mixture was stirred at room temperature for an additional 30 min, then refluxed for 2 h, the solvent was removed in vacuo, and the residue was treated with 2M HCl solution until the solution was pH 2. Then, the solution was treated with 2M sodium hydroxide solution until the pH of the solution was 14, and after removing water from the solution, the residue was treated with DCM and filtered using Celite.

After obtaining a filtrate, the solvent was removed in vacuo, and the residue _{was purified by silica gel chromatography using 1.5:15:80 NH 4} OH/MeOH/CHCl ₃ to obtain 0.015 g of compound 3 represented by the following formula 3 ( 24%) was obtained.

[Formula 3]

The analysis results of compound 3 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.79 (s, 1H), 8.72 (s, 1H), 8.21 (s, 1H), 7.88 (d, J = 7.4 Hz, 2H), 7.81 (s, 1H), 7.53 (d, J = 8.6 Hz, 1H), 6.99 (t, J = 5.2 Hz, 1H), 5.46 (s, 1H), 3.50 (q, J = 6.1 Hz, 2H), 2.51 (m, 2H), 2.23 (s, 6H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ176.56, 159.34, 153.63, 149.57, 137.75, 130.31, 130.26, 130.03, 129.89, 124.54, 122.44, 119.07, 113.49, 99.97, 52.55, 43.22, 42.29; ESI MS: m/z = 382 [M+H] ⁺ .

<Synthesis of compound 4>

Compound 4: 5-[5-[6-[2-(diethylamino)ethylamino]pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione (5-[5-[6-[2-(Diethylamino)ethylamino]pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 4 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using compound H-2 instead of compound H-1 in the synthesis of compound E.

The product is 1.5:15:80 NH ₄ OH/MeOH/CHCl ₃ And 2.5:25:80 NH ₄ OH / MeOH / CHCl ₃ was purified by silica gel chromatography to obtain a compound 4 represented by the following formula (4) in 49% yield.

[Formula 4]

The analysis results of compound 4 are as follows:

¹ H-NMR (500 MHz, CD ₃ OD): δ8.70 (s, 1H), 8.22 (s, 1H), 7.88 (s, 1H), 7.84 (s, 1H), 7.75 (dd, J = 8.6 , 1.7 Hz, 1H), 7.51 (d, J = 8.6 Hz, 1H), 3.91 (t, J = 6.3 Hz, 2H), 3.50 (t, J = 6.3 Hz, 2H), 3.28 (q, J = 1.5) Hz, 4H), 1.26 (t, J = 7.4 Hz, 7H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ159.40, 154.25, 149.68, 137.65, 131.97, 130.74, 130.15, 128.88, 124.49, 122.36, 118.93, 113.37, 99.77, 49.67, 47.15, 35.58, 8.99; ESI MS: m/z = 410 [M+H] ⁺ .

<Synthesis of compound 5>

Compound 5: 5-[5-[6-[2-(diethylamino)ethoxy]pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione (5-[5-[6-[2-(Dimethylamino)ethoxy]pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 5 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using compound H-3 instead of compound H-1 in the synthesis of compound E.

The product was purified by silica gel chromatography using 1.5:15:80 NH ₄ OH/MeOH/CHCl ₃ to obtain compound 5 represented by the following Chemical Formula 5 in 56% yield.

[Formula 5]

The analysis results of compound 5 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.97 (s, 1H), 8.76 (s, 1H), 8.72 (d, J = 1.7 Hz, 1H), 8.19 (s, 1H), 8.00 (d, J = 2.9 Hz, 1H), 7.95 (dd, J = 8.6, 1.7 Hz, 1H), 7.58 (d, J = 8.6 Hz, 1H), 4.67 (t, J = 5.4 Hz, 2H), 3.28 (t, J = 5.4 Hz, 2H), 2.65 (s, 6H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ177.17, 159.20, 158.90, 149.86, 137.85, 133.96, 132.81, 128.85, 128.58, 124.80, 122.14, 119.84, 113.34, 101.76, 61.18, 56.51, 44.18; ESI MS: m/z = 383 [M+H] ⁺ .

<Synthesis of compound 6>

compound 6: 5 -[5-[6-[3-(dimethylamino) propoxy ] pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione (5-[5-[6-[3-(Dimethylamino)propoxy]pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 6 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using compound H-4 instead of compound H-1 in the synthesis of compound E.

The product was _{purified by silica gel chromatography using 1:10:80 NH 4} OH/MeOH/CHCl ₃ to obtain compound 6 represented by the following Chemical Formula 6 in 79% yield.

[Formula 6]

The analysis results of compound 6 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.86 (s, 1H), 8.82 (d, J = 1.1 Hz, 1H), 8.76 (s, 1H), 8.15 (s, 1H), 7.93 (dd, J = 8.6, 2.3 Hz, 2H), 7.55 (d, J = 8.6 Hz, 1H), 4.50 (t, J = 6.3 Hz, 2H), 3.15 (t, J = 7.7 Hz, 2H), 2.70 (s, 6H), 2.21-2.14 (m, 2H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ177.45, 159.27, 159.24, 149.83, 137.78, 133.54, 132.66, 128.62, 127.71, 124.98, 121.78, 120.13, 113.13, 102.51, 63.19, 54.76, 43.19, 54.76, 43. 24.70; ESI MS: m/z = 397 [M+H] ⁺ .

<Synthesis of compound 7>

Compound 7: 5-[5-[6-(4-methylpiperazin-1-yl)pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione (5-[5-[6-(4-Methylpiperazin-1-yl)pyrazin-2-yl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 7 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using compound H-5 instead of compound H-1 in the synthesis of compound E.

The product was purified by silica gel chromatography using 1.5:15:80 NH ₄ OH/MeOH/CHCl ₃ to obtain compound 7 represented by the following formula 7 in 46% yield.

[Formula 7]

The analysis results of compound 7 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.25 (d, J = 1.7 Hz, 1H), 8.60 (d, J = 1.1 Hz, 1H), 8.47 (s, 1H), 8.31 (s) , 1H), 8.18 (d, J = 2.9 Hz, 1H), 7.96 (dd, J = 8.6, 1.7 Hz, 1H), 7.60 (d, J = 8.6 Hz, 1H), 3.91 (s, 4H), 3.13 (s, 5H), 2.66 (s, 3H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ176.49, 160.21, 159.34, 153.55, 149.55, 137.79, 130.43, 130.27, 130.19, 124.52, 122.45, 119.04, 113.53, 99.78, 52.21, 42.79, 42.00; ESI MS: m/z = 394 [M+H] ⁺ .

<Synthesis of compound 8>

compound 8: 5 -[5-[6-[2-(dimethylamino) ethylamino ]-2-pyradyl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione (5-[5-[6-[2-(Dimethylamino)ethylamino]-2-pyridyl]-1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 8 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using compound H-6 instead of compound H-1 in the synthesis of compound E.

The product was purified by silica gel chromatography using 1.5:15:80 NH ₄ OH/MeOH/CHCl ₃ to obtain compound 8 represented by the following formula 8 in 71% yield.

[Formula 8]

The analysis results of compound 8 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ11.83 (s, 1H), 8.70 (s, 1H), 7.94 (d, J = 2.9 Hz, 1H), 7.82 (dd, J = 8.6, 1.7 Hz, 1H), 7.51-7.45 (m, 2H), 7.09 (d, J = 7.4 Hz, 1H), 6.80 (t, J = 5.4 Hz, 1H), 6.44 (d, J = 8.0 Hz, 1H) , 3.67 (q, J = 5.9 Hz, 2H), 3.31 (t, J = 6.3 Hz, 2H), 2.81 (s, 6H); ¹³ C-NMR (125 MHz, DMSO- d ₆ ): δ177.04, 159.47, 158.55, 155.61, 138.40, 137.21, 132.89, 127.91, 124.69, 121.95, 119.14, 112.66, 108.97, 107.46, 101.43, 56.94 36.96; ESI MS: m/z = 381 [M+H] ⁺ .

<Synthesis of compound 9>

compound 9:5 -(5-phenyl--1 H -indol-3-yl) 3 H -1,3,4- oxadiazole -2- thione (5-(5-Phenyl-1 H -indol-3-yl)-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 9 was synthesized through a process similar to the synthesis method of compound 3, and was synthesized using bromobenzene instead of compound H-1 during the synthesis of compound E.

The product is 1:10:80 NH ₄ OH/MeOH/CHCl ₃ And 1.5:15:80 NH ₄ OH/MeOH/CHCl ₃ Purification by silica gel chromatography using the compound 9 represented by the following formula 9 was obtained in 55% yield.

[Formula 9]

The analysis results of compound 9 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.18 (s, 1H), 8.24 (d, J = 2.9 Hz, 1H), 8.14 (s, 1H), 7.67 (d, J = 7.4 Hz) , 2H), 7.63 (d, J = 8.6 Hz, 1H), 7.58 (dd, J = 8.3, 1.4 Hz, 1H), 7.49 (t, J = 7.7 Hz, 2H), 7.36 (t, J = 7.4 Hz) , 1H); ESI MS: m/z = 294 [M+H] ⁺ .

<Synthesis of Compound 10>

compound 10: 5 -(5-(2- Fluorophenyl )-One H -indol-3-yl) 3 H -1,3,4- oxadiazole -2-thione (5- [5- (2-Fluorophenyl) -1 H -indol-3-yl]-3 H -1,3,4-oxadiazole-2-thione)

The preparation of compound 10 was synthesized through a process similar to the synthesis method of compound 3, and in the synthesis of compound E, 2-bromofluorobenzene was used instead of compound H-1.

The product is 1:10:80 NH ₄ OH/MeOH/CHCl ₃ And 1.5:15:80 NH ₄ OH / MeOH / CHCl ₃ was purified by silica gel chromatography to obtain a compound 10 represented by the following formula (10) in 60% yield.

[Formula 10]

The analysis results of compound 10 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.29 (s, 1H), 8.33 (d, J = 2.9 Hz, 1H), 8.14 (s, 1H), 7.70 (d, J = 8.6 Hz) , 1H), 7.63 (td, J = 8.0, 1.7 Hz, 1H), 7.52 (dt, J = 8.4, 1.6 Hz, 1H), 7.50-7.46 (m, 1H), 7.40 (dd, J = 4.3, 3.2) Hz, 1H), 7.38 (d, J = 7.4 Hz, 1H); ESI MS: m/z = 312 [M+H] ⁺ .

according to Scheme 3 Pim Synthesis of novel compounds with kinase inhibitory activity

Compound 11 and Compound 12, which are S-benzyl compounds, were synthesized from Compound 2 or Compound 9 as shown in Scheme 3.

<Synthesis of compound 11>

compound 11:2 - benzylsulfanyl -5-(5- Bromo -One H -Indol-3-yl)-1,3,4- oxadiazole (2-Benzylsulfanyl-5-(5-bromo-1 H -indol-3-yl)-1,3,4-oxadiazole)

N,N-diisopropylethylamine (N,N- diisopropylethylamine; DIEA, 0.34 mmol, 0.059 mL) was added to a solution of EtOH (5.0 mL) containing the compound 2 (0.34 mmol, 0.10 g) at 0 ° C. Then, the mixture was stirred at 0° C. for 10 minutes. Benzyl bromide (benzyl bromide, 0.34 mmol, 0.037 mL) was added to the reaction mixture at room temperature, stirred for 1 hour, and the solvent was removed in vacuo. The residue _{was purified by silica gel chromatography using 1:10:80 NH 4} OH/MeOH/CHCl ₃ to obtain 0.083 g (64%) of compound 11 represented by the following Chemical Formula 11.

[Formula 11]

The analysis results of compound 11 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.22 (s, 1H), 8.19 (d, J = 2.9 Hz, 1H), 8.11 (d, J = 1.7 Hz, 1H), 7.47 (d) , J = 9.2 Hz, 1H), 7.46-7.42 (m, 2H), 7.36 (dd, J = 8.9, 2.0 Hz, 1H), 7.31 (t, J = 7.4 Hz, 2H), 7.24 (t, J = 7.4 Hz, 1H), 4.53 (s, 2H); ESI MS: m/z = 386 [M+H] ⁺ .

<Synthesis of compound 12>

compound 12:2 - benzylsulfanyl -5-(5-phenyl-1 H -Indol-3-yl)-1,3,4- oxadiazole (2-Benzylsulfanyl-5-(5-phenyl-1 H -indol-3-yl)-1,3,4-oxadiazole)

Compound 12 was prepared in the same manner as in the synthesis of Compound 11 by using Compound 9 instead of Compound 2 to obtain Compound 12 represented by the following Chemical Formula 12 in a yield of 93%.

[Formula 12]

The analysis results of compound 12 are as follows:

¹ H-NMR (500 MHz, DMSO- d ₆ ): δ12.09 (s, 1H), 8.23 (s, 1H), 8.18 (d, J = 2.9 Hz, 1H), 7.65 (d, J = 6.9 Hz) , 2H), 7.58 (d, J = 8.0 Hz, 1H), 7.53 (dd, J = 8.3, 1.4 Hz, 1H), 7.47-7.43 (m, 4H), 7.34-7.31 (m, 1H), 7.29 ( d, J = 8.0 Hz, 2H), 7.23 (t, J = 7.4 Hz, 1H), 4.53 (s, 2H); ESI MS: m/z = 384 [M+H] ⁺ .

Pim Kinase inhibitory activity measurement

The inhibitory activity of compounds against Pim-1, Pim-2 and Pim-3 kinases was measured using fluorescence polarization assay with reference to previously known methods (Lee, J., et al. , Chem. Pharm. Bull. , 62:906-914, 2014).

The IC ₅₀ value was determined by fitting data to a 4-parameter logistic function of Prism (GraphPad), and the ATP concentration was set as the Km value of Pim kinase.

_{IC 50} values for Pim kinase of compound 2, compound 9 to compound 12 compound Pim-1
IC ₅₀ (μM) Pim-2
IC ₅₀ (μM) Pim-3
IC ₅₀ (μM) compound 2 0.079 0.67 0.063 compound 9 0.26 1.24 0.46 compound 10 0.55 5.1 1.3 compound 11 >10 >10 4.7 compound 12 2.4 >10 4.9

As shown in Table 1, compound 2, a prototype of indole and 1,3,4-oxadiazole-2(3H)-thione conjugate, has inhibitory activity at submicromolar concentrations against all three Pim kinases. , and it was confirmed that it showed higher binding ability to Pim-1 and Pim-3 than to Pim-2.

Compound 9 and compound 10, which are phenyl or 2-fluorophenyl substituted compounds, showed reduced Pim kinase binding ability compared to compound 2, indicating that a bulky hydrophobic group at the 5-position of the indole ring is not suitable. means that

In addition, compounds 11 and 12 also significantly reduced Pim kinase inhibitory activity, confirming that the hydrogen bond donor of the sulfur atom is important.

_{IC 50} Values for Pim Kinase of Compounds 3 to 8 compound Pim-1
IC ₅₀ (μM) Pim-2
IC ₅₀ (μM) Pim-3
IC ₅₀ (μM) compound 3 0.006 0.23 0.007 compound 4 0.005 0.23 0.008 compound 5 0.013 0.18 0.016 compound 6 0.021 0.41 0.038 compound 7 0.033 0.45 0.024 compound 8 0.009 0.32 0.020

As shown in Table 2, in the case of Compounds 3 to 8, in which pyrazine and pyridine having an aminoalkyl substituent are attached to the 5-position of the indole ring, introduction of a heteroaromatic ring As a result, it was confirmed that it is a very potent Pim kinase inhibitor with _{a very low nanomolar IC 50} value (Pim kinase binding activity) for Pim-1 and Pim-3 kinases.

The type of linker between the aminoalkyl group and the pyrazine also affected the Pim kinase inhibitory activity, and the amino linkage had twice the binding activity to Pim-1 and Pim-3 kinases compared to the ether linkage. appeared to be

of compound 3 Pim -One Molecular docking analysis for kinases

In the present invention, in order to analyze the molecular docking of compound 3 of the present invention and Pim-1 (PDB 4DTK) kinase ATP binding site, the crystal structure of PCS-1 kinase (PDB ID: 4DTK) was obtained from the RCSB protein data bank. , Molecular docking analysis was performed using the AutoDock vina program for the structure of the compound of the present invention and the crystal structure of Pim-1 kinase.

As shown in FIG. 1 , as a result of performing molecular docking on compound 3, it was confirmed that four hydrogen bond interactions and some hydrophobic interactions may exist between compound 3 and the Pim-1 protein.

Kinase selective profile analysis of compound 4

^{In the present invention, the kinase profiler (KinaseProfiler TM} ) project of Eurofins was used to confirm the selectivity for the kinase of compound 4.

Kinase Selective Profile of Compound 4 Kinase % activity Kinase % activity Aurora-A(h) 120 JNK3(h) 74 CDK2/cyclinA (h) 89 KDR(h) 98 CK2(h) 89 MAPK2(h) 102 cSRC(h) 102 Met(h) 103 Flt3(h) 67 Plk1(h) 92 GSK3β(h) 70 SAPK2a(h) 91 IRAK4(h) 94 TAK1(h) 110 JAK2(h) 104

As shown in Table 3, Compound 4 showed a weak inhibitory effect on Flt3, GSK3β and JNK3 kinase, but _{considering the very low IC 50} value of Compound 4 for Pim-1 and Pim-3, Compound 4 was It can be seen that it has very high selectivity for Pim-1 and Pim-3.

Claims (7)

A compound represented by the following formula (1) having Pim kinase inhibitory activity:
[Formula 1]

And in the formula 1 R is a halogen, R ₁ an optionally substituted aromatic or heteroaromatic group, wherein R ₁ is an alkyl amine, a cyclic amine or an alcohol side-substituted with halogen or R _2, wherein R ₂ is an alkyl amine group am.
The compound according to claim 1, wherein the compound is represented by any one selected from the group consisting of the following formulas 2 to 10:
[Formula 2]

[Formula 3]

[Formula 4]

[Formula 5]

[Formula 6]

[Formula 7]

[Formula 8]

[Formula 9]

[Formula 10]

.
The compound according to claim 1, wherein the compound binds specifically to the ATP binding site of Pim kinase.
The compound according to claim 1, wherein the Pim kinase is Pim-1, Pim-2 or Pim-3.
A Pim kinase inhibitory composition comprising the compound having the Pim kinase inhibitory activity of any one of claims 1 to 5 as an active ingredient.
A pharmaceutical composition for preventing or treating cancer comprising the compound having the Pim kinase inhibitory activity of any one of claims 1 to 5 as an active ingredient.
A pharmaceutical composition for the treatment of diseases or disorders mediated by Pim kinase, comprising the compound having the Pim kinase inhibitory activity of any one of claims 1 to 5 as an active ingredient.

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