KR20220105425A - Thiazolopyridine or pharmaceutically acceptable salts thereof, and uses thereof - Google Patents

Abstract

The present invention relates to a thiazolopyridine compound, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof, and uses thereof. The thiazolopyridine compound, the stereoisomer, the derivative, or the pharmaceutically acceptable salt thereof can directly interact with lysyl-tRNA synthase (KRS), thereby being able to effectively inhibit the activity of KPS which induces cell migration or cancer cell metastasis without cytotoxicity. Therefore, the thiazolopyridine compound, the stereoisomer, the derivative, or the pharmaceutically acceptable salt thereof can be effectively used in anticancer drugs or cancer metastasis inhibitors.

Description

Thiazolopyridine or pharmaceutically acceptable salts thereof, and uses thereof

It relates to a thiazolopyridine compound, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof, and a use thereof.

Cancer refers to a group of abnormal cells generated by continuous division and proliferation by disrupting the balance between cell division and death due to various causes, and is also called malignant tumor. In general, cancer develops in more than 100 different parts of the body, including organs, white blood cells, bones, lymph nodes, etc. do.

Currently, as a method of treating cancer, surgery, chemotherapy, radiation therapy, etc. are used in the case of cancer detected at an early stage, but the main cause of death is induced by metastasis of cancer cells. Therefore, the identification of mechanisms that inhibit the migration or invasion of cancer cells or the development of effective therapeutic drugs are the main targets of many researchers.

Meanwhile, recently, the role of Lysyl-tRNA Synthetase (KRS) related to cell migration and especially cancer metastasis has been studied. Accordingly, KRS is emerging as a promising target for the treatment of cell migration-related diseases, particularly cancer metastasis, and efforts are being made to find a chemical inhibitor that effectively inhibits cancer cell metastasis without being toxic to target KRS.

One aspect is to provide a thiazolopyridine compound, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof.

Another aspect is to provide a pharmaceutical composition comprising the compound, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof.

Another aspect is to provide a method for treating cancer, comprising administering the compound, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof to a subject other than a human.

Hereinafter, the present invention will be described in more detail.

All technical terms used herein, unless otherwise defined, have the meanings commonly understood by one of ordinary skill in the art of the present invention. In addition, the numerical values described herein are considered to include the meaning of “about” even if not specified.

In the present specification, the term “including” is used to indicate that other components may be added and/or interposed, rather than excluding other components, unless otherwise stated.

As used herein, the term “combination thereof” means mixing or combining with one or more of the described components.

As used herein, the term “interaction” may be direct or indirect, may include a direct bond or may bind indirectly, and the binding may be mediated by another molecule.

The term "stereoisomer" refers to a compound that has the same molecular formula and the same method for linking members, but differs in spatial arrangement between atoms. The stereoisomer may be a diasteromer or an enantiomer. Enantiomers refer to isomers that do not overlap their mirror images, such as the relationship between the left and right hands, and are also called optical isomers. Enantiomers are divided into R (Rectus: clockwise) and S (sinister: counterclockwise) when 4 or more substituents differ from each other at the chiral central carbon. Diastereomers refer to stereoisomers that are not in a mirror image relationship, and can be divided into cis-trans isomers resulting from a different spatial arrangement of atoms.

The term "derivative" refers to a compound obtained by substituting a part of the structure of the compound with another atom or group of atoms.

In the present specification, a substituent is derived by exchanging one or more hydrogens with another atom or a functional group in an unsubstituted mother group. Unless otherwise stated, when a functional group is considered to be “substituted,” it means that the functional group is a halogen group, an alkyl group having 1 to 40 carbon atoms, an alkenyl group having 2 to 40 carbon atoms, or an alkynyl group having 2 to 40 carbon atoms. , means substituted with one or more substituents selected from a cycloalkyl group having 3 to 40 carbon atoms, a cycloalkenyl group having 3 to 40 carbon atoms, and an aryl group having 7 to 40 carbon atoms.

When a functional group is described as being “optionally substituted”, it is meant that the functional group may be substituted with the aforementioned substituents.

As used herein, the term "pharmaceutically acceptable salt" refers to a concentration having an effective action that is relatively non-toxic and harmless to a patient, and the side effects caused by the salt do not decrease the beneficial efficacy of the compound represented by Formula 1, etc. It means any organic or inorganic addition salt of the compound represented by . For these salts, inorganic acids and organic acids can be used as free acids, and hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, perchloric acid, phosphoric acid, etc. can be used as inorganic acids, and citric acid, acetic acid, lactic acid, maleic acid, fumarin, etc. can be used as organic acids. acid, gluconic acid, methanesulfonic acid, glycolic acid, succinic acid, tartaric acid, galacturonic acid, embonic acid, glutamic acid, aspartic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid phonic acid, 4-toluenesulfonic acid, salicylic acid, citric acid, benzoic acid or malonic acid can be used. Further, these salts include alkali metal salts (sodium salt, potassium salt, etc.) and alkaline earth metal salt (calcium salt, magnesium salt, etc.) and the like. For example, acid addition salts include acetate, aspartate, benzate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, edisylate, esylate, formate, fumarate, Gluceptate, gluconate, glucuronate, hexafluorophosphate, bibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, malate ate, malonate, mesylate, methylsulfate, naphthylate, 2-naphsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, saccharate Late, stearate, succinate, tartrate, tosylate, trifluoroacetate, aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, Potassium, sodium, tromethamine, zinc salt, etc. may be included, and among these, hydrochloride or trifluoroacetate may be included.

As used herein, the term "alkyl" means a branched or unbranched aliphatic hydrocarbon. In one embodiment, the alkyl group may be substituted or unsubstituted. The alkyl group includes a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, an isobutyl group, a tert-butyl group, a pentyl group, a hexyl group, a cyclopropyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, etc. It is not necessarily limited thereto, and each of them may be optionally substituted or unsubstituted. In one embodiment, the alkyl group may have 1 to 6 carbon atoms. For example, an alkyl group having 1 to 6 carbon atoms may be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, hexyl, etc., but is not necessarily limited thereto.

One aspect provides a thiazolopyridine compound, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof.

The thiazolopyridine may be a compound represented by Formula 1 below.

<Formula 1>

In Formula 1, R ₁ may be one selected from the group consisting of the following formula:

,

,

,

,

,

,

,

, 및

.H, CH ₃ ,

,

,

,

,

,

,

,

, and

.

In Formula 1, R ₂ is H, Me, C ₂ CH ₄ Ph(4-OMe), C ₂ CH ₄ Ph(4-NO ₂ ), CH ₂ CH ₄ Ph(4-F), and CH ₂ CH ₄ It may be one selected from the group consisting of Ph(4-Br).

The thiazolopyridine-based compound may be, for example, a compound represented by the following Chemical Formulas 2 to 17, and specifically may be a compound represented by the following Chemical Formulas 2 or 3.

<Formula 2>

<Formula 3>

<Formula 4>

<Formula 5>

<Formula 6>

<Formula 7>

<Formula 8>

<Formula 9>

<Formula 10>

<Formula 11>

<Formula 12>

<Formula 13>

<Formula 14>

<Formula 15>

<Formula 16>

<Formula 17>

The compound represented by Formula 2 is 4-(((4-Methoxyphenethyl)(thiazolo[5,4-b]pyridine-2-yl)amino)methyl)benzoic acid, or 4-(((4-methoxyphene) It may be referred to as ethyl)(thiazolo[5,4-b]pyridin-2-yl)amino)methyl)benzoic acid, and specifically, compound 4g or SL-1910 may be used in combination in the present specification.

The compound represented by Formula 3 is N-(4-methoxyphenethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide or N-(4-methoxyphenylethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide It may be referred to as zolo[5,4-b]pyridin-2-yl)acetamide, and specifically, compound 10h or SL-7620 may be used interchangeably within the present specification. For the SL-7620, the Vss (L/Kg) confirmed through the experiments in the present specification was 8.2, and the EC ₅₀ was 0.85 μM.

The compound may have an activity of directly binding to Lysyl-tRNA Synthetase (KRS) and inhibiting its activity. The compound may inhibit the activity of the KRS protein and at the same time provide activity of inhibiting cell migration, inhibiting cancer cell migration, or inhibiting cancer metastasis without cytotoxicity.

In addition, the compound may optionally have a substituent at a specific position with a halogen group, for example, may further include Cl in Formulas 2 to 17 as a substituent.

Another aspect is to provide a pharmaceutical composition for preventing or treating cancer comprising a thiazolopyridine compound, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof.

As used herein, the term "cancer" refers to or describes a physiological condition characterized by uncontrolled cell growth in mammals, which rapidly grows while infiltrating surrounding tissues and spreads or metastasizes to various parts of the body. life-threatening malignancies may include without limitation. The cancer is a carcinoma derived from epithelial cells such as lung cancer, laryngeal cancer, stomach cancer, colon/rectal cancer, liver cancer, gallbladder cancer, pancreatic cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, prostate cancer, kidney cancer, skin cancer, etc. (carcinoma), bone cancer, Connective tissue cell-derived sarcoma such as muscle cancer, adipose cancer, and fibroblast cancer; hematopoietic cell-derived cancer such as leukemia, lymphoma, and multiple myeloma; Triple negative breast cancer), brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, brain lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, nasal/sinus cancer, nasopharynx It may be at least one selected from the group consisting of head cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, chest tumor, small cell lung cancer, non-small cell lung cancer, thymus cancer, mediastinal tumor, and esophageal cancer.

The pharmaceutical composition may be one that provides cancer metastasis inhibition or epithelial-mesenchymal cell metastasis (EMT) inhibitory activity. Specifically, in the present specification, the in vivo cancer metastasis inhibitory activity of SL-1910 and SL-7620 was confirmed through a xenograft mouse model.

The pharmaceutical composition may further include a pharmaceutical additive selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, and any combination thereof.

The diluent is used to increase the amount and may be selected from the group consisting of mannitol, lactose, starch, microcrystalline cellulose, rudipress, calcium dihydrogen phosphate, and any combination thereof, but is not limited thereto. The diluent may be included in an amount of 1 to 99% by weight, preferably 20 to 80% by weight of the total weight of the solid formulation.

The binder may be selected from the group consisting of povidone, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, sodium carboxymethylcellulose, and any combination thereof, but is not limited thereto. The binder may be included in an amount of 0.5 to 15% by weight, preferably 1 to 10% by weight of the total weight of the solid formulation.

The disintegrant may be selected from the group consisting of croscarmellose sodium, crospovidone, sodium starch glycolate, and any combination thereof, but is not limited thereto. The disintegrant may be used in an amount of 1 to 30% by weight, preferably 2 to 7% by weight, based on the total weight of the solid preparation.

The lubricant is stearic acid, stearic acid metal salts (eg, calcium stearate, magnesium stearate, etc.), talc, colloidal silica, sucrose fatty acid ester, hydrogenated vegetable oil, wax, glyceryl fatty acid ester, glycerol dibehenate, and may be selected from the group consisting of any combination thereof, but is not limited thereto. The lubricant may be included in an amount of 0.3 to 7% by weight, preferably 0.5 to 5% by weight of the total weight of the solid formulation.

The pharmaceutical composition may contain about 0.1 to 500 mg of the active ingredient, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof, as a free base per unit dosage form, and from 0.5 to 500 mg based on the total weight of the preparation. It may be contained in a proportion of 50% by weight, specifically 1 to 40% by weight. The compound may be included in the pharmaceutical composition in an amount sufficient to achieve its efficacy or activity. A person skilled in the art can select and practice the upper or lower limit of the quantity of the compound contained in the composition within an appropriate range.

The pharmaceutical composition may be for oral or parenteral use, and the pharmaceutical composition may be administered orally or parenterally. It may be formulated as an oral or parenteral dosage form.

When it is prepared as the oral dosage form, it may be prepared according to any oral solid preparation known in the art, specifically, a method for preparing granules, pellets, capsules, or tablets. Oral dosage forms may be granules, powders, solutions, tablets, capsules, dry syrups, or a combination thereof. The parenteral dosage form may be an injection or an external preparation for skin. The external preparation for skin may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, or a combination thereof. In the case of parenteral administration, for example, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, endothelial administration, local administration, intranasal administration, intrapulmonary administration, rectal administration, etc. can be administered. In addition, the composition may be administered by any device capable of transporting the active agent to a target cell.

Another aspect is to provide a method for treating cancer, comprising administering the compound, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof to a subject other than a human.

The composition may be administered in combination with drugs such as steroids, anti-inflammatory agents, and antibiotics in order to effectively prevent or treat cancer or a secondary disease caused therefrom in an individual. The combined administration may be administered to the subject sequentially, simultaneously, or separately.

The administration may be 0.001 μg/mL to 1,000 mg/mL, for example, 0.01 μg/mL to 500 mg/mL, or 0.1 μg/mL to 150 mg/mL of the compound per subject per day. Administration may be administered once a day, or may be administered in several divided doses. The dosage of the pharmaceutical composition varies depending on the condition and weight of the patient, the severity of the disease, the drug form, the route of administration, and the duration, but may be appropriately selected by those skilled in the art.

The thiazolopyridine compound, a stereoisomer, derivative, or pharmaceutically acceptable salt thereof according to an aspect can directly interact with lysyl-tRNA synthase (KRS), and thus there is no cytotoxicity, but cell migration or cancer It can effectively inhibit the activity of KRS that induces cell metastasis. Accordingly, one aspect of the thiazolopyridine compound, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof can be effectively utilized in an anticancer agent or a cancer metastasis inhibitor.

1 is an agent showing anti-metastatic activity and a road confirming its activity, a diagram confirming the substituent of the agent exhibiting KRS-selective cell migration inhibitory activity (FIG. 1A), tetracycline inducibility showing a high level of human KRS T52D mutation Results showing the KRS-selective cell migration inhibitory activity of SL-1910 and its analogs at 3 μM against MDA-MB-231 (FIG. 1B) and the concentration-dependent cell migration inhibitory effect of SL 1910 on the MDA-MB-231 cell line It is a figure showing the confirmed result (FIG. 1C).
2 is a diagram showing the results for YH16899 (FIG. 2A) and SL-1910 (FIG. 2B) as a road confirming the fluorescence-based binding analysis for Kd value measurement. Each experiment was carried out twice.
3 is a road confirming the 2D-NMR binding study between SL-1910, compound 4g, and KRS protein, in the absence (black) and presence (red) of 0.2 mM ¹⁵ N labeled KRS ₁ in the absence (black) and presence (red) of 0.4 mM SL-1910. As a result of confirming the two-dimensional ¹ H ¹⁵ N _TROSY spectrum of -207, the NMR cross peaks of the ¹ H ¹⁵ N TROSY spectrum from the Y112, H120, R161, K190 and T191 residues are displayed at the bottom (Fig. 3A) and As a result of confirming the results showing chemical shift disturbance (CSP) (Fig. 3B), the binding site of the perturbed residue of the anticodon binding domain based on CSP was mapped to the result expressed by the backbone model of KRS in the presence of SL-1910. Regarding the results, the results of indicating the strongly and weakly perturbed residues in red and orange, respectively (Fig. 3C), the results of confirming the docking position of the S-form of YH16899 (blue) (Fig. 3D), and the docking of SL-1910 (green) It is a figure shown to the result of confirming the position (FIG. 3E). CSP-determined residues are indicated in red and orange (PDB ID: 3BJU).
Figure 4 is a road showing the effect of measuring the cancer metastasis inhibitory effect of SL-1910 and SL-7620 in vivo. The result of visually confirming the group (FIG. 4A) and the result of comparing the number of metastatic lung cancer crystals in the control group and the SL-1910 and SL-7620 treatment groups (FIG. 4B) and confirming the measured body weight after drug administration (FIG. 4C) )to be.

Hereinafter, the present invention will be described in more detail through experimental examples and examples. However, these Experimental Examples and Examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these Experimental Examples and Examples.

Experimental Example 1. Experimental materials and methods

All commercial reagents and solvents were purchased from commercial suppliers and used. Tetrahydrofuran was distilled from sodium benzophenone ketyl, and dichloromethane, acetonitrile, triethylamine and pyridine were freshly distilled with calcium hydride. Flash column chromatography was performed using silica gel 60 (230-400 mesh, Merck) and preparative thin layer chromatography was used with glass-supported silica gel plates (1 mm, Merck). Thin layer chromatography was performed to monitor the reaction. All reactions were performed in a dry argon atmosphere in flame-dried glassware. ¹ H NMR and ¹³ C NMR spectra were recorded on a Bruker Avance III HD (800 MHz, using 5 mm CPTCI CryoProbe) spectrometer. Chemical shifts are expressed in ppm (δ) downfield in tetramethylsilane (internal standard) with coupling constants in hertz (Hz). Multiplicity was denoted by abbreviations such as singlet (s), doublet (d), doublet of doublet (dd), triplet (t), quartet (q), multiplet (m) and broad (br). . Mass spectra and HRMS were recorded with a VG Trio 2 GC-MS instrument and a JEOL JMS AX, respectively. All analogs were purified to at least 95% purity. The purity of the analogs was determined by reverse phase high performance liquid chromatography (HPLC) (Shimadzu, LC 20AD liquid chromatograph, waters/sunfire ^™ C18 5 μm (4.6×150 mm)).

Experimental Example 2. Cell migration analysis

Cell migration assays were performed using a CytoSelect 96-well cell migration system (pore size of 8.0 μm, Cell Biolabs) according to the manufacturer's instructions. Briefly, the human breast cancer MDA-MB-231 tetracycline-inducible cell line for high-level expression of human KRS T52D mutants was used in the assay.

Cells treated with 2 μg/ml doxycycline for 3 days were suspended in serum-free DMEM medium. Inhibitor compound containing YH21931 as a positive control was mixed with the cell suspension and added to the upper chamber of a 96-well cell transfer plate at a density of 1× ^{10 4} cells per well. Serum-free medium containing 10 μg/ml laminin (Sigma) was added to the wells of the feeder tray as an attractant. Cells were allowed to migrate at 37 °C for 4 h, and migratory cells were detached from the underside of the membrane using cell detachment solution, lysed with lysis buffer and stained with CyQuant GR dye solution. Fluorescence intensity was measured at 480 nm/520 nm with a fluorescence plate reader. Data are presented as triplicate mean±standard deviation (SD).

Experimental Example 3. Protein Preparation

¹⁵ N-labeled KRS _1-207 with an N-terminal hexahistidine tag in the pET28a vector was E. coli Bl21 (DE3) in M9 minimal medium enriched with ¹⁵ NH ₄ Cl (99% ¹⁵ N; cambridge Isotope Labolatories) as the sole nitrogen source. ) was overexpressed. After 0.5 mM isopropyl β-D-1 thiogalactopyranoside (IPTG) was added to express the protein at an OD ₆₀₀ of 0.5 to 0.6, the cells were grown at 37° C. for 3 hours while shaking. Cells were harvested by centrifugation at 2,970xg for 30 min at 4°C, resuspended in lysis buffer containing 20mM HEPES pH 7.0, 500mM NaCl, 10% glycerol, and 1mM PMSF, and disrupted by intermittent sonication on ice. did. The removed supernatant was separated from the lysate by centrifugation at 32,500 x g for 60 min at 4 °C, applied to a 5 ml HisTrap (GE Healthcare) column pre-equilibrated with lysis buffer, and 20 mM HEPES pH 7.0, 500 mM NaCl and 50 mM imidazole were added. It was washed with buffer A containing His-tagged proteins were eluted with buffer A containing 500 mM imidazole. His-tags were cleaved by incubation with thrombin at 4° C. overnight during dialysis against PMSF-free buffer 20 mM HEPES pH 7.0, buffer containing 500 mM NaCl, and His-tags were removed from proteins by passing through a HisTrap column. After concentration by ultrafiltration (Millipore), the protein was loaded onto a Superdex 200 16/60 gel filtration column (GE Healthcare) pre-equilibrated with 20 mM HEPES pH 7.0, 150 mM NaCl and 7 mM B-ME (beta-mercaptoethanol). loaded.

Experimental Example 4. NMR analysis

Buffer containing 20 mM HEPES (pH 7.0), 150 mM NaCl, 7 mM B ME and 0.8% DMSO in the presence or absence of 0.4 mM SL10 at triple resonance 298 K with buffer using an Avance 600 MHz NMR spectrometer equipped with a probe (Bruker). 0.2 mM ¹⁵ N-labeled KRS _1-207 ¹ H- ¹⁵ N TROSY (transverse relaxation optimized spectroscopy) experiments were performed. Chemical shift perturbation (CSP) of ¹⁵ N and ¹ H nuclei was analyzed by overlaying the ¹ H- ¹⁵ N TROSY spectrum of the free protein with the spectrum with SL10. The magnitude of the bound ¹⁵ N- ¹ H chemical shift perturbation (δ, ppm) was calculated based on the equation, where H and N represent the change in proton ( ¹ H) and nitrogen ( ¹⁵ N) chemical shifts, respectively.

Experimental Example 5. Confirmation of molecular docking

The anticodon domain structure of LysRS (PDB ID: 3BJU) was prepared using Gasteiger Charge, the protein structure remained robust upon docking, and the SL10 binding site was defined from CSP-mapped residues obtained from NMR binding experiments. Grid dimensions with 65X65X69 points and 0.375 Å grid spacing were used for sampling of ligand conformation at CSP-based binding sites. The SL10 chemical derivative was modeled with the SYBY LX 2.0 molecular modeling package (http://tripos.com) and vacuum using the Powell algorithm and Tripos force field for 5000 iterations along a termination gradient of 0.05 kcal/(mol Å). Energy was minimized with the Gasteiger Huckel charge set in the dielectric environment. An energy-minimized SL10 derivative was prepared in Autodock and a Gasteiger charge was assigned to the chemical. 200 docking poses of the ligand were sampled using Autodock4.2 (http://autodock.scripps.edu/) public domain software. Ligand conformation was sampled by Lamarckian genetic algorithm, parameters set to 200 independent runs, an initial population of 150 randomly placed, 2.5 X 10 ⁶ energy assessments, up to 27000 repetitions, mutation rate 0.02, cross It was set by an over rate of 0.80 and an elitism value. The docking pose of the most frequent cluster, which is a low-energy pose, was selected for analysis. Pymol ( http://www.pymol.org ) was used for distance analysis.

Experimental Example 6. Confirmation of metastasis in 4T1 cells of breast cancer in mice

To determine the degree of metastasis in the 4T1 breast cancer cell line, 4T1 cells (4 x 10 ⁴ ) were surgically injected into 2 mammary fat pads of anesthetized 7-week-old female BALB / cAnCr mice. Primary tumors were removed within 10 days after inoculation (primary tumor volume: 100-150 mm ³ ), and mice having a uniform distribution of resected primary tumors were treated with control group and SL-1910 and SL-7620 at 100 mg/kg. (8 animals in each group) were divided into experiments. SL-1910, SL-7620 and vehicle (corn oil: polyethylene glycol 400: Tween 80: methyl cellulose (1%) = 20: 30: 1: 49) were orally administered once a day starting on day 1 after tumor resection. . All mice were sacrificed, and lungs were excised 28 days after injection of 4T1 cells. Lungs were fixed in Bouin's solution and metastasis was evaluated by counting lung nodules. All animals were treated in accordance with WOJUNGBIO's Animal Care and Use Committee guidelines with approved animal study protocols.

Example 1. Synthesis of N,N-dialkylthiazolo[5,4-b]pyridin-2-amine (N,N-dialkylthiazolo[5,4-b]pyridin-2-amine: SL-1910)

1.1 Synthesis of N,N-dialkylthiazolo[5,4-b]pyridin-2-amine (N,N-dialkylthiazolo[5,4-b]pyridin-2-amine: SL-1910) and its analogs

The synthesis of SL-1910 and its analogs, which consists of a bicyclic core and a tertiary amine moiety, was synthesized through a three-step sequence. Microwave assisted aromatic nucleophilic substitution was performed at a later stage for efficient synthesis of core scaffold hopping derivatives. Thus prepared N,N-dialkylthiazolo[5,4-b]pyridin-2-amine (N,N-dialkylthiazolo[5,4-b]pyridin-2-amine: SL-1910), including its The reaction scheme for the derivative compound is as follows:

<Reaction Scheme 1>

1.2 Reaction for Reductive Alkylation

To synthesize amine 1 of Scheme 1, NaHB(OAc) ₃ (2 eq) was added to a solution of 4-methoxyphenethylamine (2.5 eq.) and or 4-formylbenzonitrile, 1 eq.) in MeOH at 0 ° C. was added in The reaction mixture was warmed to room temperature and stirred until the reaction was complete. The reaction mixture was extracted with ethyl acetate. The combined organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified using flash column chromatography on silica gel to give the corresponding secondary amines 2a or 2b of Scheme 1.

1.3 Aromatic Nucleophilic Substitution and Induction of Reaction for Hydrolysis

1.5 equivalents of 2a or 2b and 3a-1 (1 equivalent) in toluene of Scheme 1 were added, and to the solution 5 equivalents of TEA was added at room temperature. The reaction mixture was refluxed until the reaction was complete and concentrated under reduced pressure. The residue was dissolved in water and brine was added. The mixture was extracted with ethyl acetate. The combined organic layers were dried over MgSO ₄ and concentrated under reduced pressure to give the corresponding crude benzoate or benzonitrile. To dissolve 1 equivalent of the benzoate or benzonitrile in 0.25 M MeOH, 10 equivalents of 4N aqueous NaOH solution was added and the mixture was heated to 60°C. After completion of the reaction, the reaction mixture was extracted with ethyl acetate. The combined organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified using flash column chromatography on silica gel to give the corresponding amine derivative 4a-1.

1.4 Synthesis of target material and analysis of the material structure of the synthesized compound

1.4.1. Methyl 4-(((4-methoxyphenethyl)amino)methyl)benzoate (2a of Scheme 1)

The coupling reaction of Scheme 1 of amine 1 (47mL, 320.8mmol) and methyl 4-formylbenzoate (20.6g, 125.5mmol) was reacted with NaHB(OAc) ₃ (53.1g, 251.0mmol) in the presence of the reducibility of the above example. It was carried out according to the reaction procedure for alkylation. Flash column chromatography of the crude product on silica gel (EtOAc/n-Hexane = 1:3) gave 34.9 g (93%) of 2a. ¹ H-NMR (CDCl ₃ , 800 MHz) δ 7.96 (d, J = 8.2 Hz, 2H), 7.36 (d, J = 8.0 Hz, 2H), 7.09 (d, J = 8.5 Hz, 2H), 6.81 (d) , J = 8.4 Hz, 2H), 3.88 (s, 3H), 3.85 (s, 2H), 3.77 (s, 3H), 2.85 (t, J = 7.2 Hz, 2H), 2.78 (t, J = 7.1 Hz) , 2H).

1.4.2. Methyl 4-(((4-methoxyphenethyl)amino)methyl)benzonitrile (2b of Scheme 1)

The coupling reaction of Scheme 1 of amine 1 (1.3mL, 8.6mmol) and 4-formylbenzonitrile (500mg, 3.4mmol) was subjected to the reductive alkylation of the above example in the presence of NaHB(OAc) ₃ (1.5g, 6.9 mmol). It was carried out according to the reaction procedure for Flash column chromatography of the crude product on silica gel (EtOAc/n-Hexane = 1:3) gave 819.7 mg (93%) of 2b. ¹ H-NMR (CDCl ₃ , 800 MHz) δ 7.57 (d, J = 8.1 Hz, 2H), 7.38 (d, J = 8.1 Hz, 2H), 7.09 (d, J = 8.5 Hz, 2H), 6.82 (d , J = 8.4 Hz, 2H), 3.83 (s, 2H), 3.77 (s, 3H), 2.83 (t, J = 7.0 Hz, 2H), 2.75 (t, J = 6.9 Hz, 2H).

1.4.3. 4-(((4-methoxyphenethyl)(5,6,7-trifluorobenzo[d]thiazol-2-yl)amino)methyl)benzoic acid ( FIG. 1 4a )

In the presence of TEA (1.5 mL, 11 mmol), chloride 3a (487 mg, 2.2 mmol) and amine 2a (718 mg, 2.4 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a 4 N NaOH solution added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 15: 1) gave 330 mg (32% in 2 steps) of 4a: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.03 (d , J= 8.1 Hz, 2H), 7.31 (d, J= 8.0 Hz, 2H), 7.15 (dd, J= 10.8, 5.5 Hz, 1H), 7.10 -7.05 (m, 2H), 6.83 (d, J= 8.3 Hz, 2H), 4.67 (s, 2H), 3.77 (s, 3H), 3.64 (t, J= 7.5 Hz, 2H), 2.92 (t, J= 7.6 Hz, 2H); 13C NMR (200 MHz, CDCl3) δ171.20, 169.98, 168.87, 148.48, 142.35, 131.09, 130.75 (2 carbons), 129.93, 129.77 (2 carbons), 128.73, 127.84, 127.52 (2 carbons), 114.24, 114.21 ( 2 carbons), 113.71, 102.54, 55.29, 54.68, 53.63, 32.64; HR-MS (FAB) calculated 473.1147 for C ₂₄ H ₂₀ N ₂ O 3 S (M+ H ⁺ ), found 473.1136.

1.4.4. 2-((4-carboxybenzyl)(4-methoxyphenethyl)amino)benzo[d]thiazole-6-carboxylic acid (4b in FIG. 1)

In the presence of TEA (0.1 mL, 0.5 mmol), chloride 3b (21 mg, 0.1 mmol) and amine 2a (32 mg, 0.1 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a 4 N NaOH solution added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 14 mg (31% in 2 steps) of 4b: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.30 (s) , 1H), 7.96 (d, J = 8.4 Hz, 1H), 7.84 (d, J = 8.3 Hz, 2H), 7.49 (d, J = 8.1 Hz, 1H), 7.38 (d, J = 8.4 Hz, 2H) ), 7.14 (d, J = 8.7 Hz, 2H), 6.84 (d, J = 8.5 Hz, 2H), 4.78 (s, 2H), 3.77-3.71 (m, 5H), 3.74-2.95 (t, J = 7.5 Hz, 2H); ¹³ C NMR (200 MHz, CD ₃ OD) δ 172.87, 172.79, 164.07, 163.89, 163.72, 160.84, 157.98, 142.92, 135.22, 132.49 (2 carbons), 131.77 (2 carbons), 129.99 (2 carbons), 129.42 ( 2 carbons), 124.75, 119.56, 115.93 (2 carbons), 56.46, 55.59, 50.65, 34.38; HR MS (FAB) calculated for C ₂₅ H ₂₂ N ₂ O ₅ S(M) 462.1249, found 462.1247.

1.4.5. 4-(((6-(hydroxymethyl)benzo[d]thiazol-2-yl)(4-methoxyphenethyl)amino)methyl)benzoic acid (4c in FIG. 1 )

In the presence of TEA (0.03 mL, 0.3 mmol), bromide 3c (13 mg, 0.1 mmol) and amine 2a (18 mg, 0.1 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a 4 N NaOH solution added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 15 mg (62% in 2 steps) of 4c: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 7.95 (d , J = 8.2 Hz, 2H), 7.64 (d, J = 1.8 Hz, 1H), 7.46 (d, J = 8.1 Hz, 1H), 7.31 (d, J = 8.1 Hz, 2H), 7.28 (dd, J) = 8.3, 1.7 Hz, 1H), 7.12 (d, J = 8.7 Hz, 2H), 6.83 (d, J = 8.6 Hz, 2H), 4.72 (s, 2H), 4.62 (s, 2H), 3.74 (s) , 3H), 3.69 (t, J = 7.5 Hz, 2H), 2.92 (t, J = 7.5 Hz, 2H); ¹³ C NMR (200 MHz, CD 3 OD) δ 170.99, 166.23, 160.77, 153.94, 142.97, 137.12, 132.64, 132.52, 131.83 (2 carbons), 131.81, 131.75 (2 carbons), 129.05 (2 carbons), 127.47, 121.47, 120.03, 115.89 (2 carbons), 66.03, 56.46, 56.26, 55.33, 34.42.; HR MS (FAB) calculated 449.1535 for C ₂₅ H ₂₅ N ₂ O ₄ S (M+ H ⁺ ), found 449.1533.

1.4.6. 4-((benzo [d] oxazol-2-yl (4-methoxyphenethyl) amino) methyl) benzoic acid (4d in Figure 1)

In the presence of TEA (0.1 mL, 0.5 mmol), chloride 3d (15 mg, 0.1 mmol) and amine 2a (32 mg, 0.1 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a 4 N NaOH solution added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 28 mg (71% in 2 steps) of 4d: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.02 (d , J= 7.9 Hz, 2H), 7.42 (d, J= 7.8 Hz, 1H), 7.32 (d, J= 8.0 Hz, 2H), 7.27 (d, J= 7.9 Hz, 1H), 7.18 (t, J) = 7.7 Hz, 1H), 7.08 (d, J= 8.1 Hz, 2H), 7.04 (t, J= 7.7 Hz, 1H), 6.80 (d, J= 8.1 Hz, 2H), 4.68 (s, 2H), 3.75 (s, 3H), 3.70 (t, J = 7.5 Hz, 2H), 2.92 (t, J = 7.4 Hz, 2H); ¹³ C NMR (200 MHz, CD ₃ OD) , 115.79 (2 carbons), 110.77, 56.39, 56.38, 53.79, 52.31, 34.90; HR-MS (FAB) calculated for C ₂₄ H ₂₃ N ₂ O ₄ S (M+ H ⁺ ) 403.1658, found 403.1664.

1.4.7. 4-(((4-methoxyphenethyl)(5-nitrobenzo[d]oxazol-2-yl)amino)methyl)benzoic acid (4e in FIG. 1)

In the presence of TEA (1.8 mL, 13 mmol), chloride 3e (500 mg, 2.5 mmol) and amine 2b (738 mg, 2.8 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 20: 1) gave 530 mg (47% in 2 steps) of 4e: 1H-NMR (CDCl ₃ , 800 MHz) δ8.22 (d , J= 2.2 Hz, 1H), 8.01 (dd, J= 8.6, 2.3 Hz, 1H), 7.77 (d, J= 8.1 Hz, 2H), 7.37 -7.26 (m, 3H), 7.07 (d, J= 8.5 Hz, 2H), 6.81 (d, J= 8.3 Hz, 2H), 4.68 (s, 2H), 3.75 (s, 3H), 3.71 (t, J= 7.4 Hz, 2H), 2.92 (t, J= 7.4 Hz, 2H); ¹³ C NMR (200 MHz, CDCl ₃ ) δ168.62, 163.87, 158.51, 152.70, 145.34, 143.78, 140.24, 133.01, 129.88, 129.77 (2 carbons), 127.98 (2 carbons), 127.90 (2 carbons), 117.30, 114.16 (2 carbons), 111.85, 108.52, 55.29, 52.33, 50.29, 33.26.; HR-MS (FAB) calculated for C ₂₄ H ₂₁ N ₃ O ₆ S (M) 447.1430, found 447.1446.

1.4.8. 4-(((5,7-difluoro-1H-benzo[d]imidazol-2-yl)(4-methoxyphenethyl)amino)methyl)benzoic acid (4f in FIG. 1)

In the presence of TEA (0.1 mL, 1 mmol), chloride 3f (27 mg, 0.1 mmol) and amine 2a (47 mg, 0.2 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 18 mg (29% in 2 steps) of 4f: ¹ H-NMR (CD ₃ OD ₃ , 800 MHz) δ7. 95 (d, J= 8.0 Hz, 2H), 7.28 (d, J= 7.8 Hz, 2H), 7.12 (d, J= 8.7 Hz, 2H), 6.81 (d, J= 8.5 Hz, 2H), 6.78 ( dd, J= 8.8, 2.3 Hz, 1H), 6.60 (td, J= 10.4, 2.2 Hz, 1H), 4.65 (s, 2H), 3.73 (s, 3H), 3.65 (t, J= 7.4 Hz, 2H) ), 2.89 (t, J=7.3 Hz, 2H); ¹³ C NMR (200 MHz, CD ₃ OD) δ170.13, 164.07, 163.90, 163.72, 160.69, 160.40, 159.10, 158.85, 143.89, 132.88, 131.87 (2 carbons), 131.80 (2 carbons), 131.51,128.96 (2 carbons), 115.78 (2 carbons), 97.12, 56.44, 53.93, 52.33, 34.76.; HR-MS (FAB) calculated for C ₂₄ H ₂₁ F ₂ N ₃ O ₃ S (M + H ⁺ ) 438.1629, found 438.1630.

1.4.9. 4-(((4-methoxyphenethyl)(thiazolo[5,4-b]pyridin-2-yl)amino)methyl)benzoic acid (4g in Fig. 1, SL-1910)

In the presence of TEA (6.8 mL, 48.9 mmol), 3 g (1.7 mg, 9.8 mmol) of chloride of Scheme 1 and amine 2a (3.2 mg, 10.8 mmol) were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 15: 1) gave 2.9 mg (70% in 2 steps) of 4 g: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.22 ( d, J= 4.8 Hz, 1H), 8.04 (d, J= 8.2 Hz, 2H), 7.76 (dd, J= 8.1, 1.5 Hz, 1H), 7.31 (d, J= 8.1 Hz, 2H), 7.26 - 7.22 (m, 1H), 7.08 (d, J= 8.7 Hz, 2H), 6.82 (d, J= 8.6 Hz, 2H), 4.71 (s, 2H), 3.76 (s, 3H), 3.68 (t, J) = 7.4 Hz, 2H), 2.93 (t, J=7.5 Hz, 2H); ¹³ C NMR (200 MHz, CDCl ₃ ) δ170.47, 167.14, 158.44, 154.94, 147.34, 142.11, 141.82, 130.61 (2 carbons), 130.05, 129.77 (2 carbons), 129.36, 127.45 (2 carbons), 125.11, 121.41, 114.15 (2 carbons), 55.24, 54.02, 52.98, 32.65; HR-MS (FAB) calculated for C ₂₃ H ₂₂ N ₃ O ₃ S (M + H ⁺ ) 420.1382, found 420.1384.

1.4.10. 4-(((4-methoxyphenethyl)(thiazolo[4,5-b]pyridin-2-yl)amino)methyl)benzoic acid (4h in FIG. 1 )

In the presence of TEA (0.1 mL, 0.6 mmol), chloride 3h (20.0 mg, 0.1 mmol) and amine 2a (38.9 mg, 0.1 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 15 : 1) gave 26 mg (54% in 2 steps) of 4h: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.47 -8.37 (m, 1H), 8.01 (d, J= 7.7 Hz, 2H), 7.86 (d, J= 7.8 Hz, 1H), 7.38 -7.27 (m, 2H), 7.08 (d, J= 7.9 Hz, 2H) , 6.97 (dd, J= 6.6, 4.0 Hz, 1H), 6.80 (d, J= 8.1 Hz, 2H), 4.72 (s, 2H), 3.75 (s, 3H), 3.69 (s, 2H), 2.93 ( s, 2H); ¹³ C NMR (200 MHz, CDCl3) δ170.16, 169.63, 164.15, 158.45, 146.47, 141.89, 130.61 (2 carbons), 130.07, 129.83 (2 carbons), 129.48, 129.17, 127.64 (2 carbons), 125.27, 116.34 , 114.15 (2 carbons), 55.26, 54.47, 53.23, 32.64; HR-MS (FAB) calculated for C ₂₃ H ₂₂ N ₃ O ₃ S (M + H ⁺ ) 420.1382, found 420.1368.

1.4.11 4-(((4-methoxyphenethyl)(thiazolo[4,5-c]pyridin-2-yl)amino)methyl)benzoic acid (4i in FIG. 1)

In the presence of TEA (0.1 mL, 0.6 mmol), chloride 3i (20.0 mg, 0.1 mmol) and amine 2a (38.9 mg, 0.1 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 15: 1) gave 10 mg (21% in 2 steps) of 4i: ¹ H-NMR (CD ₃ OD, 800 MHz) δ 8.64 (s, 1H), 8.13 (d, J= 5.3 Hz, 1H), 7.95 (d, J= 7.9 Hz, 2H), 7.75 (d, J= 5.3 Hz, 1H), 7.31 (d, J= 8.1 Hz) , 2H), 7.12 (d, J= 8.6 Hz, 2H), 6.83 (d, J= 8.5 Hz, 2H), 4.77 (s, 2H), 3.79 -3.68 (m, 5H), 2.93 (t, J= 7.4 Hz, 2H); ¹³ C NMR (200 MHz, CD ₃ OD) δ171.46, 160.83, 152.21, 142.64, 141.66, 141.54, 140.49, 137.60, 132.44 (2 carbons), 131.78 (2 carbons), 130.24, 128.98 (2 carbons), 127.33 , 118.53, 115.91 (2 carbons), 56.46 (2 carbons), 55.60, 34.31; HR-MS (FAB) calculated for C ₂₃ H ₂₂ N ₃ O ₃ S (M + H ⁺ ) 420.1382, found 420.1386.

1.4.12. 4-(((6-chlorothiazolo[5,4-b]pyridin-2-yl)(4-methoxyphenethyl)amino)methyl)benzoic acid ( 1 of 4j)

In the presence of TEA (0.1 mL, 0.9 mmol), chloride 3j (35.0 mg, 0.2 mmol) and amine 2a (56.9 mg, 0.2 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 46 mg (59% in 2 steps) of 4j: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.14 ( d, J= 2.1 Hz, 1H), 8.03 (d, J= 8.2 Hz, 2H), 7.72 (d, J= 2.1 Hz, 1H), 7.31 (d, J= 8.1 Hz, 2H), 7.08 (d, J= 8.2 Hz, 2H), 6.83 (d, J= 8.4 Hz, 2H), 4.70 (s, 2H), 3.77 (s, 3H), 3.67 (t, J= 7.5 Hz, 2H), 2.92 (t, J=7.5 Hz, 2H); ¹³ C NMR (200 MHz, CDCl3) δ169.55, 168.26, 158.55, 153.19, 147.72, 142.13, 140.72, 130.74 (2 carbons), 129.93, 129.85, 129.78 (2 carbons), 128.79, 127.56 (2 carbons), 124.55 , 114.22 (2 carbons), 55.28, 54.02, 53.17, 32.67; HR-MS (FAB) calculated 454.0992 for C ₂₃ H ₂₁ N ₃ O ₃ SCl (M + H ⁺ ), found 454.0998.

1.4.13. 4-(((4-methoxyphenethyl)(thiazolo[5,4-d]pyrimidin-2-yl)amino)methyl)benzoic acid (4k in FIG. 1 )

In the presence of TEA (0.2 mL, 1.37 mmol), chloride 3k (47.0 mg, 0.3 mmol) and amine 2a (89.8 mg, 0.3 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 56 mg (49% in 2 steps) of 4k: ¹ H-NMR (CD ₃ OD, 800 MHz) δ 8.68 (s, 1H), 8.66 (s, 1H), 7.79 (d, J= 8.3 Hz, 2H), 7.39 (d, J= 8.1 Hz, 2H), 7.13 (d, J= 8.6 Hz, 2H), 6.83 (dd, J=6.7, 1.9 Hz, 2H), 4.80 (s, 2H), 3.82-3.77 (m, 2H), 3.73 (s, 3H), 2.96 (t, J=7.3 Hz, 2H); ¹³ C NMR (200 MHz, CD3OD) δ 169.72, 166.09, 164.12, 163.97, 163.51, 160.91, 151.57, 148.06, 144.93, 132.24, 132.05 (2 carbons), 131.84 (2 carbons), 129.36 (2 carbons), 115.95 2 carbons), 65.55, 56.45, 34.30, 31.47; HR-MS (FAB) calculated 421.1334 for C ₂₂ H ₂₁ N ₄ O ₃ S (M + H ⁺ ), found 421.1344.

1.4.14. 4-(((4-methoxyphenethyl)(oxazolo[5,4-b]pyridin-2-yl)amino)methyl)benzoic acid (41 in FIG. 1 )

In the presence of TEA (0.2 mL, 1.3 mmol), chloride 31 (41.0 mg, 0.3 mmol) of Scheme 1 and amine 2a (87.4 mg, 0.3 mmol) were carried out according to the aromatic nucleophilic substitution procedure of the above example. The resulting methyl ester was hydrolyzed with a solution of 4N-NaOH added to MeOH according to standard procedures. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10: 1) gave 36 mg (34% in 2 steps) of 4l: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 7.93 ( dd,J= 5.2, 1.7 Hz, 1H), 7.75 (d, J= 8.2 Hz, 2H), 7.58 (dd, J= 7.7, 1.5 Hz, 1H), 7.31 (d, J= 8.1 Hz, 2H), 7.13 (dd, J= 7.6, 5.0 Hz, 1H), 7.07 (d, J= 8.5 Hz, 2H), 6.80 (d, J= 8.6 Hz, 2H), 4.65 (s, 2H), 3.76 (s, 3H) ), 3.68 (t, J=7.4 Hz, 2H), 2.91 (t, J=7.4 Hz, 2H); ¹³ C NMR (200 MHz, CDCl ₃ ) δ168.78, 161.61, 158.41, 158.38, 140.59, 138.87, 135.98, 132.88, 130.11, 129.80 (2 carbons), 127.89 (2 carbons), 127.87 (2 carbons), 123.18, 120.65, 114.11 (2 carbons), 55.27, 51.91, 49.97, 33.29; HR-MS (FAB) calcd for C ₂₃ H ₂₁ N ₃ O ₄ (M) 403.1532, found 403.1540.

1.4.15. 4-(((4-methoxyphenethyl)(6-nitrobenzo[d]oxazol-2-yl)amino)methyl)benzonitrile (5 in FIG. 1)

In the presence of TEA (0.3 mL, 2.3 mmol), chloride 3e (300 mg, 1.5 mmol) and amine 2b (335.3 mg, 1.3 mmol) of Scheme 1 were carried out according to the aromatic nucleophilic substitution procedure of the above example. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated, dissolved in water and brine was added. The mixture was extracted with a mixture of IPA and chloroform (1:4). The organic layer was dried over MgSO ₄ and concentrated. Flash column chromatography of the crude product on silica gel (EtOAc / n-Hexane = 1:3) gave 403 mg (62%) of compound 5: ¹ H-NMR (CDCl ₃ , 800 MHz) δ 8.20 (d, J = 2.2 Hz, 1H), 8.01 (dd, J= 8.7, 2.3 Hz, 1H), 7.61 (d, J= 8.1 Hz, 2H), 7.34 (d, J= 8.0 Hz, 2H), 7.31 (d, J) = 8.8 Hz, 1H), 7.07 (d, J= 8.5 Hz, 2H), 6.81 (d, J= 8.4 Hz, 2H), 4.66 (s, 2H), 3.76 (s, 3H), 3.71 (t, J) = 7.3 Hz, 2H), 2.93 (t, J= 7.3 Hz, 2H).

1.4.16. 4-(((6-aminobenzo[d]oxazol-2-yl)(4-methoxyphenethyl)amino)methyl)benzonitrile (6 in FIG. 1)

SnCl ₂ · 2H ₂ O (263 mg, 1.2 mmol) in an ethanol solution containing the nitrobenzoxazole 5 compound of Reaction Scheme 1 (100 mg, 0.2 mmol). The reaction mixture was refluxed for 3 h and concentrated under reduced pressure. The residue was dissolved in CH ₂ Cl ₂ and 2N-NaOH was added. The organic layer was washed with brine and dried over MgSO ₄ . Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 20: 1) gave 61 mg (62%) of compound 6: ¹ H NMR (CDCl ₃ , 800 MHz) δ 7.59 (dd, J = 8.3 , 4.6 Hz, 2H), 7.44 (s, 1H), 7.34 7.31 (m, 2H), 7.23 7.19 (m, 1H), 7.11 7.07 (m, 1H), 7.06 (d, J = 6.4 Hz, 2H), 6.81 (dd, J = 8.5, 4.1 Hz, 2H), 4.65 (s, 2H), 3.75 (s, 3H), 3.67 (t, J = 7.3 Hz, 2H), 2.90 (t, J = 7.3) Hz, 2H).

1.4.17. 4-(((4-methoxyphenethyl)(5-(2-(4-methoxyphenyl)acetamido)benzo[d]oxazol-2-yl)amino)methyl)benzoic acid ( FIG. 1 7a )

DIPEA in a solution containing aminobenzoxazole 6 (13.7 mg, 0.03 mmol), 4-methoxyphenylacetic acid (4.8 mg, 0.03 mmol) and HATU (10.1 mg, 0.03 mmol) of Scheme 1 and dry CH ₂ Cl ₂ (0.01 mL, 0.1 mmol)) was added. The reaction mixture was stirred at 40 °C until the reaction was complete and diluted with CH ₂ Cl ₂ . The mixture was washed with saturated aqueous NaHCO ₃ solution. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. 13 mg (79 %) of N-(2-((4-cyanobenzyl)(4-methoxyphenethyl)amino by flash column chromatography of the crude product on silica gel (EtOAc / n-Hexane = 1:1) ) Benzo[d]oxazol-5-yl)2-(4-methoxyphenyl)acetamide was obtained: ¹ H NMR (CDCl ₃ , 800 MHz) δ 7.57 (d, J = 7.9 Hz, 2H), 7.40 (d, J = 1.8Hz, 1H ), 7.31 (d, J = 8.0Hz, 2H), 7.27 7.23 (m, 2H), 7.15 7.10 (m, 2H), 7.08 (s, 1H), 7.05 (d, J = 8.5 Hz, 2H), 6.92 (d, J = 8.5 Hz, 2H), 6.80 (d, J = 8.4 Hz, 2H), 4.6 1 (s, 2H), 3.81 (s, 3H), 3.76 (s) , 3H), 3.68 (s, 2H), 3.65 (t, J = 7.5 Hz, 2H), 2.89 (t, J = 7.3 Hz, 2H). The acetamide was hydrolyzed with MeOH containing 4N-NaOH according to standard procedures to obtain 2 mg (14%) of compound 7a: ¹ H-NMR (DMSO-d ₆ , 800 MHz) δ7.83 (d , J= 8.2 Hz, 2H), 7.60 (d, J= 2.1 Hz, 1H), 7.37 (d, J= 8.1 Hz, 2H), 7.33 (s, 1H), 7.28 (d, J= 8.6 Hz, 1H) ), 7.25 (d, J= 8.6 Hz, 2H), 7.16 (dd, J= 1.9 Hz, 1H), 7.15 (d, J= 8.8 Hz, 2H), 6.89 (d, J= 8.6 Hz, 2H), 6.84 (d, J= 8.4 Hz, 2H), 4.73 (s, 2H), 3.73 (s, 3H), 3.69 (s, 3H), 3.65 (t, J= 7.6 Hz, 2H), 3.55 (s, 2H) ), 2.86 (t, J=7.6 Hz, 2H); ¹³ C NMR (200 MHz, DMSO-d6) δ169.11, 167.57, 162.62, 157.99, 157.80, 144.53, 143.33, 140.62, 135.77, 133.42, 130.35, 130.07 (2 carbons), 129.72 (2 carbons), 128.00, 127.79 (2 carbons), 127.12 (2 carbons), 113.83 (2 carbons), 113.71 (2 carbons), 111.64, 108.43, 106.99, 55.01, 54.95, 51.06, 42.41, 40.41, 32.47.; HR-MS (FAB) calculated for C ₃₃ H ₃₁ N ₃ O ₆ (M) 565.2213, found 565.2205.

1.4.18. 4-(((((4-methoxyphenethyl)(5-(4-(4-methylpiperazin-1-yl)benzamido)benzo[d]oxazol-2-yl)amino)methyl) benzoic acid (7b in Fig. 1)

To aminobenzoxazole 6 (32.7 mg, 0.1 mmol), 4-(4-methylpiperazin-1-yl)benzoic acid (18.1 mg, 0.1 mmol), HOBt (13.3 mg, 0.1 mmol) and DMF of Scheme 1 DIPEA (0.01 mL, 0.1 mmol) was added to a solution containing the added EDC·HCl (15.7 mg, 0.1 mmol) at 0 ° C. The reaction mixture was stirred at room temperature until the reaction was completed and diluted with CH ₂ Cl ₂ The organic layer was dried over MgSO ₄ and concentrated under reduced pressure.Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 10:1) gave 23 mg (48%) of N-(2-((4) -cyanobenzyl)(4-methoxyphenethyl)amino)benzo[d]oxazol-5-yl)4-(4-methylpiperazin-1-yl)benzamide was obtained: ¹ H-NMR (CDCl ₃ , 800 MHz) δ7.74 (d, J= 8.0 Hz, 2H), 7.57 (s, 1H), 7.55 (d, J= 7.9 Hz, 2H), 7.35 -7.30 (m, 1H), 7.30 (d, J= 7.9 Hz, 2H), 7.19 (d, J= 8.4 Hz, 1H), 7.04 (d, J= 8.2 Hz, 2H), 6.89 -6.82 (m, 2H), 6.78 (d, J= 8.1 Hz, 2H), 4.60 (s, 2H), 3.74 (s, 3H), 3.70 -3.53 (m, 2H), 3.42 -3.22 (br s, 4H), 2.92 -2.84 (m, 2H), 2.83 - 2.67 (br s, 4H), 2.47 (s, 3H).

The benzamide was hydrolyzed with MeOH containing 4N-NaOH according to standard procedures to obtain 3 mg (12%) of compound 7b: 1H-NMR (CDOD, 800 MHz) δ7.90 (d, J = 8.7 Hz, 2H), 7.85 (d, J= 8.1 Hz, 2H), 7.68 (d, J= 2.0 Hz, 1H), 7.39 (d, J= 8.0 Hz, 2H), 7.31 (dd, J= 8.5) , 2.1 Hz, 1H), 7.28 (d, J= 8.6 Hz, 1H), 7.13 (d, J= 8.5 Hz, 2H), 7.08 (d, J= 8.8 Hz, 2H), 6.81 (d, J= 8.7) Hz, 2H), 4.73 (s, 2H), 3.76 -3.69 (m, 5H), 3.54 -3.43 (br s, 4H), 3.17 -3.01 (br s, 4H), 2.92 (t, J= 7.4 Hz, 2H), 2.72 (s, 3H); 13 C NMR (200 MHz, DMSO-d ₆ ) δ167.59, 164.85, 162.57, 157.81, 152.97, 144.67, 143.21, 140.67, 135.94, 133.44, 130.38, 129.74 (2 carbons), 128.99 (2 carbons), 127.81 (2 carbons) carbons), 127.15 (2 carbons), 123.79, 113.84 (2 carbons), 113.48 (2 carbons), 112.98, 108.33, 108.23, 54.97, 54.37 (2 carbons), 51.09, 49.89 (2 carbons), 46.96, 45.75, 32.50 .; HR-MS (FAB) calculated for C ₃₆ H ₃₇ N ₅ O ₅ (M) 619.2795, found 619.2800.

1.4.19. 2-((4-carboxybenzyl)(4-methoxyphenethyl)amino)thiazolo[5,4-b]pyridine4-oxide (8 in FIG. 1)

mCPBA (26.4mg, 0.1mmol) was added to a solution of 4g (30mg, 0.01mmol) of the compound dissolved in CH ₂ Cl ₂ at room temperature. After the reaction suspension became clear and colorless to pale yellow, the mixture was stirred until the reaction was complete. The reaction mixture was diluted with CH ₂ Cl ₂ and washed with saturated aqueous NaHCO ₃ solution. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Flash column chromatography of the crude product on silica gel (CH ₂ Cl ₂ /MeOH = 16:1) gave 24 mg (76%) of compound 8: ¹ H-NMR (CD ₃ OD, 800 MHz) δ8.06 (d, J= 6.3 Hz, 1H), 7.99 (d, J= 8.4 Hz, 2H), 7.59 (d, J= 8.3 Hz, 1H), 7.42 (dd, J= 8.2, 6.3 Hz, 1H), 7.39 (d, J= 8.1 Hz, 2H), 7.14 (d, J= 8.6 Hz, 2H), 6.83 (d, J= 8.5 Hz, 2H), 4.83 (s, 2H), 3.83 -3.75 (br s, 2H) ), 3.74 (s, 3H), 2.97 (t, J=7.3 Hz, 2H); ¹³ C NMR (200 MHz, CD ₃ OD) δ170.97, 169.67, 160.93, 152.80, 145.53, 142.88, 133.69, 133.48, 132.14, 132.06 (2 carbons), 131.84 (2 carbons), 129.39 (2 carbons), 125.31 , 120.74, 115.98 (2 carbons), 56.46, 55.83, 50.65, 34.22.; HR-MS (FAB) calculated for C ₂₃ H ₂₂ N ₃ O ₄ S (M + H ⁺ ) 436.1331, found 436.1320.

Example 2. Synthesis of N-(4-methoxyphenylethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide: SL-7620) and analogs thereof

2.1 Synthesis of analogues of SL-7620

An analog of SL-7620 composed of a bicyclic core and a tertiary amine moiety was synthesized by the following method. Microwave assisted aromatic nucleophilic substitution was performed at a later stage for efficient synthesis of core scaffold hopping derivatives. Unless specifically described in this synthesis, the present synthesis method followed the synthesis example of Example 1. The reaction scheme for SL-7620 and its derivative compounds thus prepared is as follows:

<Reaction Scheme 2>

The reagents and reaction conditions used in steps a and b in Reaction Scheme 2, respectively, are as follows. Step a is halides, DIPEA, Cs ₂ CO ₃ , acetonitrile at 70 °C, or halides, DIPEA, CH ₂ Cl ₂ at room temperature. In step b, each compound of Table 1 below was synthesized by adding NaOH, H ₂ O, and MeOH at 60 °C.

2.2 Synthesis of target material and analysis of the material structure of the synthesized compound

In Chemical Formula 1, a compound, which is a target material compound in which R ₁ and R ₂ are the following functional groups, was synthesized according to the reaction scheme and the synthesis method described below, and the material structure thereof was analyzed.

[Table 1]

2.2.1. N-(4-Methoxyphenylethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide (compound 10h, SL-7620)

To prepare a solution of 1 equivalent of N-(4-methoxyphenethyl)thiazolo[5,4-b]pyridin-2-amine in anhydrous acetonitrile, 1.5 equivalents of cesium carbonate and 1 equivalent of acetyl chloride was added at 0 °C. The reaction mixture was warmed to room temperature and stirred until the reaction was complete. The reaction mixture was extracted with ethyl acetate. The combined organic layers were dried over MgSO4 and concentrated under reduced pressure. The residue was purified using flash column chromatography on silica gel to give compound 10h, N-(4-methoxyphenylethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide. were: ¹ H NMR (800 MHz, CDCl3) δ 8.50 (dd, J = 4.7, 1.5 Hz, 1H), 8.06 (dd, J = 8.1, 1.5 Hz, 1H), 7.36 (dd, J = 8.1, 4.6 Hz, 1H), 7.13 - 7.09 (m, 2H), 6.87 - 6.83 (m, 2H), 4.40 (t, J = 7.3 Hz, 2H), 3.78 (s, 3H), 3.07 (t, J = 7.3 Hz, 2H) ), 2.15 (s, 3H); ¹³ C NMR (201 MHz, CDCl3) δ 171.03, 158.61, 156.87, 145.84, 141.67, 129.97, 129.95, 127.92, 121.05, 114.26, 55.28, 49.95, 33.49, 29.69, 22.94; Calculated for HR-MS (FAB) C17H17N3O2S (M + H ⁺ ) 328.1114, found 328.1115.

2.2.1. Structural analysis of analogs of N-(4-methoxyphenylethyl)-N-(thiazolo[5,4-b]pyridin-2-yl)acetamide (compound 10h, SL-7620)

The results of analyzing the structure of the synthesized compound 9 are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.18 (dd, J = 4.8, 1.5 Hz, 1H), 8.00 - 7.95 (m, 2H), 7.77 - 7.74 (m, 1H), 7.31 - 7.27 (m, 2H), 7.24 (q, J = 4.7 Hz, 1H), 7.09 - 7.05 (m, 2H), 6.84 - 6.80 (m, 2H), 4.69 (s) , 2H), 3.88 (s, 3H), 3.76 (s, 3H), 3.66 (t, J = 7.5 Hz, 2H), 2.91 (t, J = 7.5 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 206.91, 167.09, 166.63, 166.15, 158.45, 154.82, 147.37, 141.43, 130.07 (2C), 129.75 (2C), 129.72, 127.45 (2C), 125.15, 121.38 (125.15, 121.38) 2C), 55.24, 53.98, 52.97, 52.13, 32.64; HR-MS (FAB) calculated 434.1533 for C ₂₄ H ₂₄ N ₃ O ₃ S ⁺ (M + H ⁺ ), found 434.1534.

The results of analyzing the structure of the synthesized compound 10a are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.23 (d, J = 5.0 Hz, 1H), 7.96 (d, J = 8.2 Hz, 1H), 7.63 - 7.59 (m, 2H), 7.43 (d, J = 6.8 Hz, 1H), 7.31 (d, J = 8.1 Hz, 2H), 7.09 - 7.05 (m, 2H), 6.85 - 6.81 (m, 2H) , 4.70 (s, 2H), 3.78 (s, 3H), 3.71 (d, J = 7.8 Hz, 2H), 2.95 (t, J = 7.3 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 167.65, 158.70, 142.77, 132.72 (2C), 129.77 (2C), 129.29, 128.10 (2C), 124.81, 122.07, 118.35, 114.34 (2C), 112.09, 70.26, 67 , 58.01, 55.30, 53.73, 32.64, 30.17; HR-MS (FAB) calculated for C ₂₃ H ₂₁ N ₄ OS ⁺ (M + H ⁺ ) 401.1431, found 401.1443.

The results of analyzing the structure of the synthesized compound 11 are as follows: ¹ H NMR (800 MHz, DMSO) δ 8.40 (t, J = 5.4 Hz, 1H), 8.08 (dd, J = 4.7, 1.5 Hz, 1H) , 7.67 (dd, J = 8.0, 1.5 Hz, 1H), 7.24 (dd, J = 8.0, 4.7 Hz, 1H), 7.20 - 7.16 (m, 2H), 6.89 - 6.85 (m, 2H), 3.72 (s) , 3H), 3.58 (td, J = 7.3, 5.4 Hz, 2H), 2.85 (t, J = 7.3 Hz, 2H); ¹³ C NMR (201 MHz, DMSO) δ 164.69, 157.75, 154.82, 146.53, 141.54, 130.98, 129.67 (2C), 123.61, 121.12, 113.78 (2C), 54.97, 45.04, 33.65; calculated for HR-MS (FAB) C ₁₅ H ₁₆ N ₃ OS ⁺ (M + H ⁺ ) 286.1009, found 286.1009.

The results of analyzing the structure of the synthesized compound 10b are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.14 (dd, J = 4.7, 1.5 Hz, 1H), 7.69 (dd, J = 8.1, 1.5 Hz) , 1H), 7.18 (dd, J = 8.0, 4.7 Hz, 1H), 7.15 - 7.11 (m, 2H), 6.85 - 6.80 (m, 2H), 3.76 (s, 3H), 3.71 (t, J = 7.5) Hz, 2H), 3.08 (s, 3H), 2.92 (dd, J = 8.4, 6.7 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 166.80, 158.35, 155.62, 147.37, 141.75, 129.78 (2C), 124.33, 121.11, 114.08 (2C), 113.73, 55.22, 38.44, 32.64, 29.66; HR-MS (FAB) calculated 300.1165 for C ₁₆ H ₁₈ N ₃ OS ⁺ (M + H ⁺ ), found 300.1171.

The results of analyzing the structure of the synthesized compound 10c are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.53 (dd, J = 4.8, 1.4 Hz, 1H), 8.10 (dd, J = 8.1, 1.5 Hz, 1H), 7.48 (t, J = 7.6 Hz, 1H), 7.42 - 7.38 (m, 3H), 7.27 - 7.23 (m, 2H), 6.82 (d, J = 8.2 Hz, 2H), 6.74 (d, J) = 8.2 Hz, 2H), 4.42 (t, J = 7.3 Hz, 2H), 3.76 (s, 3H), 2.97 (t, J = 7.3 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 171.68, 159.21, 158.46, 156.75, 146.05, 141.84, 134.37, 130.72, 129.98 (2C), 129.75, 128.55 (2C), 128.08, 126.87 (2C), 114.03 (2C), 114.03 (2C) 2C), 55.28, 51.20, 33.51; HR-MS (FAB) calculated for C ₂₂ H ₂₀ N ₃ O ₂ S ⁺ (M + H ⁺ ) 390.1271, found 390.1279.

The results of analyzing the structure of the synthesized compound 10d are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.56 (dd, J = 4.7, 1.5 Hz, 1H), 8.12 (dd, J = 8.1, 1.5 Hz, 1H), 7.64 (d, J = 8.2 Hz, 2H), 7.42 (dd, J = 8.1, 4.7 Hz, 1H), 7.19 (d, J = 8.1 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.75 (d, J = 8.6 Hz, 2H), 4.38 (t, J = 6.7 Hz, 2H), 3.78 (s, 3H), 2.98 (t, J = 6.7 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 169.67, 158.73, 158.60, 156.60, 146.49, 141.73, 138.43, 132.27 (2C), 130.17 (2C), 129.41, 128.44, 127.55 (2C), 121.42, 114.35,74, 127 114.21 (2C), 55.35, 51.27, 33.34; HR-MS (FAB) calculated 415.1223 for C ₂₃ H ₁₉ N ₄ O ₂ S ⁺ (M + H ⁺ ), found 415.1232.

The results of analyzing the structure of the synthesized compound 10e are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.52 (dd, J = 4.7, 1.5 Hz, 1H), 8.09 (dd, J = 8.1, 1.5 Hz, 1H), 7.38 (dd, J = 8.1, 4.7 Hz, 1H), 7.19 (q, J = 8.1 Hz, 4H), 6.87 - 6.83 (m, 2H), 6.76 - 6.72 (m, 2H), 4.44 (dd , J = 8.1, 6.5 Hz, 2H), 3.75 (s, 3H), 2.98 (t, J = 7.3 Hz, 2H), 2.39 (s, 3H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 171.86, 159.44, 158.40, 156.71, 145.92, 141.90, 141.20, 131.40, 129.97 (2C), 129.82 (2C), 129.14, 128.00 (2C), 127.05, 113.97 ( 2C), 55.26, 51.22, 33.49, 21.50; HR-MS (FAB) calculated for C ₂₃ H ₂₂ N ₃ O ₂ S ⁺ (M + H ⁺ ) 404.1427, found 404.1437.

The results of analyzing the structure of the synthesized compound 10 f are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.54 (dd, J = 4.7, 1.5 Hz, 1H), 8.11 (dd, J = 8.1, 1.6 Hz) , 1H), 7.49 - 7.44 (m, 1H), 7.40 (dd, J = 8.1, 4.7 Hz, 1H), 7.16 (td, J = 7.5, 1.0 Hz, 1H), 7.13 (t, J = 8.9 Hz, 1H), 7.03 - 6.99 (m, 1H), 6.80 (d, J = 8.5 Hz, 2H), 6.75 - 6.72 (m, 2H), 4.35 (q, J = 7.2 Hz, 2H), 3.75 (s, 3H) ), 2.97 (t, J = 7.2 Hz, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 167.22, 158.47, 158.32, 157.08, 156.59, 146.20, 141.71, 132.48, 129.94 (2C), 129.73, 129.10, 128.22, 124.74, 122.83, 121.22, 115.07 () , 55.28, 50.78, 33.39; HR-MS (FAB) calculated 408.1177 for C ₂₂ H ₁₉ FN ₃ O ₂ S ⁺ (M + H ⁺ ), found 408.1179.

The results of analyzing the structure of 10 g of the synthesized compound are as follows: ¹ H NMR (800 MHz, CDCl ₃ ) δ 8.52 (dd, J = 4.6, 1.5 Hz, 1H), 8.10 (dd, J = 8.1, 1.5 Hz) , 1H), 7.62 (dd, J = 1.7, 0.9 Hz, 1H), 7.38 (dd, J = 8.1, 4.6 Hz, 1H), 7.31 (dd, J = 3.6, 0.9 Hz, 1H), 7.20 - 7.16 ( m, 2H), 6.85 - 6.81 (m, 2H), 6.58 (dd, J = 3.5, 1.7 Hz, 1H), 4.82 - 4.78 (m, 2H), 3.77 (s, 3H), 3.19 - 3.14 (m, 2H); ¹³ C NMR (201 MHz, CDCl ₃ ) δ 159.57, 159.30, 158.41, 156.81, 146.57, 146.03, 145.43, 141.72, 130.20, 129.82 (2C), 128.13, 121.12, 120.68, 114.04 (2C), 112.42, 55.28, 49.72 , 34.23; HR-MS (FAB) calculated 380.1063 for C ₂₀ H ₁₈ N ₃ O ₃ S ⁺ (M + H ⁺ ), found 380.1066.

The results of analyzing the structure of the synthesized compound 12 are as follows: ¹ H NMR (800 MHz, DMSO) δ 9.03 (s, 1H), 8.13 (dd, J = 4.8, 1.5 Hz, 1H), 7.95 - 7.91 (m) , 2H), 7.70 (dd, J = 8.0, 1.5 Hz, 1H), 7.51 - 7.47 (m, 2H), 7.27 (dd, J = 8.1, 4.8 Hz, 1H), 4.71 (d, J = 4.8 Hz, 2H); ¹³ C NMR (201 MHz, DMSO) δ 167.09, 165.00, 154.44, 146.11, 143.68, 141.68, 129.60, 129.52 (2C), 127.34 (2C), 124.13, 121.32, 46.31; HR-MS (FAB) calculated 286.0645 for C ₁₄ H ₁₂ N ₃ O ₂ S ⁺ (M + H ⁺ ), found 286.0647.

The results of analyzing the structure of the synthesized compound 13a are as follows: ¹ H NMR (800 MHz, DMSO) δ 8.15 (dd, J = 4.7, 1.5 Hz, 1H), 7.93 (dd, J = 8.5, 2.1 Hz, 2H) ), 7.76 (dd, J = 8.0, 1.5 Hz, 1H), 7.42 (d, J = 8.1 Hz, 2H), 7.31 (dd, J = 8.1, 4.8 Hz, 1H), 4.91 (s, 2H), 3.18 (s, 3H); ¹³ C NMR (201 MHz, DMSO) δ 166.60, 155.03, 146.80, 141.84, 129.73 (2C), 129.36, 127.32 (2C), 124.35, 121.77, 121.62, 115.36, 56.95, 38.13; HR-MS (FAB) calculated for C ₁₅ H ₁₄ N ₃ O ₂ S ⁺ (M + H ⁺ ) 300.0801, found 300.0808.

The results of analyzing the structure of the synthesized compound 13b are as follows: ¹ H NMR (800 MHz, DMSO) δ 8.18 - 8.13 (m, 3H), 7.92 (s, 2H), 7.77 (dd, J = 8.1, 1.5 Hz) , 1H), 7.59 - 7.55 (m, 2H), 7.42 (d, J = 8.1 Hz, 2H), 7.31 (dd, J = 8.1, 4.7 Hz, 1H), 4.87 (s, 2H), 3.86 (t, J = 7.3 Hz, 2H), 3.15 (t, J = 7.5 Hz, 2H); ¹³ C NMR (201 MHz, DMSO) δ 166.13, 154.88, 146.84, 146.49, 146.24, 142.06, 141.61, 130.27 (2C), 130.23, 129.66 (2C), 127.28 (2C), 126.02, 124.55, 123.52 (2C) 121.60, 62.00, 51.89, 32.6; HR-MS (FAB) calculated for C ₂₂ H ₁₉ N ₄ O ₄ S ⁺ (M + H ⁺ ) 435.1122, found 435.1126.

The results of analyzing the structure of the synthesized compound 13c are as follows: ¹ H NMR (800 MHz, DMSO) δ 8.15 (dd, J = 4.7, 1.5 Hz, 1H), 7.94 - 7.90 (m, 2H), 7.76 (dd , J = 8.1, 1.5 Hz, 1H), 7.43 (d, J = 8.3 Hz, 2H), 7.31 (dd, J = 8.1, 5.0 Hz, 3H), 7.16 - 7.07 (m, 2H), 4.87 (s, 2H), 3.80 - 3.73 (m, 2H), 2.97 (t, J = 7.6 Hz, 2H); ¹³ C NMR (201 MHz, DMSO) δ 167.04, 166.18, 161.60, 160.40, 154.82, 146.56, 141.91, 134.51, 130.72 (2C), 129.91, 129.68 (2C), 127.36 (2C), 124.50, 121.60, 115.23 (2C) ), 62.00, 52.67, 31.93; HR-MS (FAB) calculated 408.1177 for C ₂₂ H ₁₉ FN ₃ O ₂ S ⁺ (M + H ⁺ ), found 408.1180.

The results of analyzing the structure of the synthesized compound 13d are as follows: ¹ H NMR (800 MHz, DMSO) δ 8.15 (dd, J = 4.8, 1.5 Hz, 1H), 7.91 (d, J = 8.3 Hz, 2H), 7.77 (dd, J = 8.1, 1.5 Hz, 1H), 7.48 (d, J = 8.3 Hz, 2H), 7.43 (d, J = 8.2 Hz, 2H), 7.31 (dd, J = 8.1, 4.8 Hz, 1H) ), 7.25 (d, J = 8.4 Hz, 2H), 4.87 (s, 1H), 3.81 - 3.73 (m, 2H), 2.96 (t, J = 7.6 Hz, 2H); ¹³ C NMR (201 MHz, DMSO) δ 167.03, 166.16, 154.81, 146.56, 141.93, 137.81, 131.31 (2C), 131.16 (2C), 129.92, 129.69 (2C), 129.59, 127.36 (2C), 121.62,55, 121.62,55 119.56, 62.00, 51.53, 32.15; HR-MS (FAB) calculated for C ₂₂ H ₁₉ BrN ₃ O ₂ S ⁺ (M + H ⁺ ) 468.0376, found 468.0387.

Example 3. Confirmation of cancer metastasis inhibitory activity of SL-1910 and SL-7620

3.1 Confirmation of cancer metastasis inhibitory activity of SL-1910

In order to confirm that the synthesized bicyclic analogue having a tertiary amine branch and having a tertiary amine branch, such as SL-1903, inhibits KRS-selective cell migration and exhibits metastasis inhibitory activity, a human KRS T52D mutant triple-negative KRS-dependent migration inhibitory activity was confirmed using the human breast cancer cell line, MDA-MB-231 tetracycline-induced cell line, and the experimental results are shown in FIG. 1 .

As confirmed in FIGS. 1A and 1B , the compound having a 2-aminotrifluorobenzothiazole core with SL-1903 corresponding to the parent compound (4a) showed about 28% inhibition of cancer metastasis at 3 μM. confirmed that. On the other hand, it was confirmed that YH16899 had significantly lower metastasis inhibitory activity at the same concentration. Through these results, it was confirmed that the flexible form of the 2-aminobenzothiazole core induced a more intimate interaction with the main residue compared with the iminobenzothiazole core. It was confirmed that benzothiazole analogues having other substituents, such as compounds 4b and 4c, had almost no migration inhibitory activity, whereas compounds 4d, 4e, 7a and 7b, which are benzoxazole analogues, exhibited moderate cell migration inhibitory activity. .

4-methoxyphenylacetamido or 4-(4-methylpiperazin-1-yl)benzo at position 5 of the benzoxazole core (7a and 7b) in anticipation of further occupancy in the hydrophobic pocket composed of Glu195 and Gln 155 An amido group substituent was introduced. As shown in FIG. 1B , it was confirmed that the analog compound 7a having a hydrophobic 4-methoxyphenylacetamido group showed excellent cell migration inhibitory activity, whereas the compound 7b having a piperazine group showed low inhibitory activity. In addition, it was confirmed that analog 4f composed of benzimidazole instead of benzothiazole exhibited low cell migration inhibitory activity. In particular, compound 4g (SL-1910) containing thiazolo[5,4-b]pyridine instead of benzothiazole core exhibited about 70% cell migration inhibitory activity at 3 μM, indicating that it may have the best cancer metastasis inhibitory activity. Confirmed. It was confirmed that this result appears because it is closely related to the docking score of SL-1910 (19.26), which is higher than that of YH16899 or SL-1903. Compounds 4h and 4i with thiazolo[4,5-b]pyridine and thiazolo[4,5-c]pyridine (isomeric analogues of SL-1910) inhibited cell migration less than SL-1903 containing benzothiazole It was confirmed that the activity was shown. These results show that sulfur in thiazole can exhibit an important positioning effect, and it can be confirmed that nitrogen in pyridine also exhibits an important positioning effect. Accordingly, in compound 4j having a sulfur and nitrogen position similar to that of compound 4g, as confirmed in FIG. 1B, even when a chloride substituent is added to the 6-position of thiazolo[5,4-b]pyridine, the cell migration inhibitory activity is 4g and It can be seen that they remain the same. The N-oxide analogue of compound 8 with thiazolo[5,4-b]pyridine and thiazolo[5, 4-d] Compound 4k having a pyrimidine core did not show any cell migration inhibitory effect at 3 μM. In addition, it was confirmed that the analog compound 4l composed of oxazolopyridine, a hybrid of thiazolopyridine (Compound 4g, SL-1910) and benzoxazole (Compound 4d), exhibited some cell migration inhibitory effect.

In addition, as confirmed in FIG. 1C , it was confirmed that compound 4g, SL-1910, had an EC50 value of 81 nM for the KRS mutant MDA-MB-231 cell line, which was higher than the previously known EC50 value of 8.5 μM for the compound YH16899 H226 cell line. It was confirmed that it was 100 times or more of the drug efficacy.

Overall, it was confirmed that compound 4g, SL-1910, exhibited the most excellent anti-cancer activity inhibiting activity.

3.2 Confirmation of cancer metastasis inhibitory activity of SL-7620

By changing only the compound in the same method and conditions as in Example 3.1, the cancer metastasis inhibitory activity of the compound including SL-7620 was specifically confirmed and shown in Table 2.

[Table 2]

As confirmed in Table 2 above, similar to SL-1910 confirmed in Example 3.1, compound 5h, SL-7620, exhibited more than 70% cell migration inhibitory activity when treated with 3 μM, thereby having the best cancer metastasis inhibitory activity. confirmed that there is.

Therefore, it was confirmed that the most excellent anticancer activity inhibiting activity was observed in SL-1910, which is compound 4g, and SL-7620, which is 10h, and thus the possibility of being utilized as an anticancer agent or cancer metastasis inhibitor was confirmed.

Example 4. Identification of binding sites between SL-1910 and KRS protein

In addition, a fluorescence-based binding titration experiment was performed to specifically confirm the binding activity of SL-1910, which exhibits the cancer metastasis inhibitory activity, to human KRS protein, and the results of the confirmation are shown in FIG. 2 , specifically YH16899. The binding analysis is shown in FIG. 2A and the result of confirming the binding analysis of SL-1910 is shown in FIG. 2B. The Kd values for YH16899 and SL-1910 binding to human KRS protein were confirmed by measuring changes in fluorescence emission according to binding to KRS at 310 nm, 365 nm and 445 nm, respectively.

As confirmed in FIGS. 2A and 2B , the Kd value of SL-1910 was 5.75 μM, which was about 9 times lower than the Kd value of 49.24 μM of YH16899. Through this, it was specifically confirmed that compound 4g, SL-1910, exhibited a much higher binding affinity for human KRS protein than the conventionally known YH16899.

Example 5. Confirmation of docking mechanism for KRS protein of SL-1910

To confirm the docking mechanism of thiazolopyridine to KRS of SL-1910 with a high bicyclic core, 2D-NMR binding analysis was performed. Through the above analysis, the binding mechanism between SL-1910 and human KRS protein, including identification of interaction residues, is shown in FIG. 3 . It was confirmed that the directional moiety of YH16899 is constituted by a hydrophobic pocket formed by the side chains of Leu142, Phe144, Met157, Thr191 and Ile199. It is known that this non-polar pocket is further bent by the polar side chains of His120, Asn159, Arg161 and Lys192 to form an optimal site for protein binding and drug targeting.

3A, 3B and 3C, SL-1910 was confirmed to interact with Tyr 112, Gly 118 and His 120, confirming that the same binding pocket as YH16899 appeared. In particular, as confirmed in FIGS. 3D and 3E, it was confirmed that YH16899 and SL-1910 showed similar binding patterns through a docking simulation based on 2D-NMR binding analysis. In particular, compared to YH16899, SL-1910 has a flexible tertiary amine structure in the middle of the molecule, which is predicted to interact with additional amino acid residues including 161Arg, 190Lys and 191Thr. It was confirmed that a more excellent cancer cell migration inhibitory effect was exhibited.

Example 6. Confirmation of actual cancer metastasis inhibitory activity in mouse models of SL-1910 and SL-7620

In order to confirm whether the compounds 4g, SL-1910 and SL-7620, which were identified in the above Examples, exhibit substantial therapeutic effects, an in vivo cancer metastasis inhibition assay using a xenograft mouse model was performed. Specifically, in vivo cancer cell anti-metastatic activity of SL-1910 and SL-7620 in a mouse 4T1 xenograft metastasis model was investigated, and this is shown in FIG. 4 .

In the group subjected to oral administration of SL-1910 and SL-7620 at a dose of 100 mg/kg, as confirmed in FIGS. 4A and 4B, the number of metastatic lung cancer nodules was reduced by about half in the group treated with SL-1910 than in the control group. could In particular, it was confirmed that the group treated with SL-7620 showed significant anti-metastatic activity that could reduce the number of metastatic lung cancer nodules compared to the group treated with SL-1910. In addition, as confirmed in FIG. 4C , it was confirmed that SL-1910 and SL-7620 had no detectable toxicity compared to the control group.

Therefore, overall, it was confirmed that SL-1910 and SL-7620 showed excellent anti-metastatic activity against cancer cells while being non-toxic, so that they could be widely used as cancer cell metastasis inhibitors.

Claims (10)

A compound represented by the following formula (1), a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof:
<Formula 1>

In Formula 1, R ₁ is one selected from the group consisting of the following formula,
H, CH ₃ ,

,

,

,

,

,

,

,

, and

,
In Formula 1, R ₂ is H, Me, C ₂ CH ₄ Ph(4-OMe), C ₂ CH ₄ Ph(4-NO ₂ ), CH ₂ CH ₄ Ph(4-F), and CH ₂ CH ₄ one selected from the group consisting of Ph(4-Br). The method according to claim 1, wherein the compound is a compound represented by Formula 2 or 3, a stereoisomer, derivative, or pharmaceutically acceptable salt thereof:
<Formula 2>

<Formula 3>

. The method according to claim 1, wherein the compound is lysyl-tRNA synthase (Lysyl-tRNA Synthetase: KRS) is a compound having an activity to inhibit its activity by binding directly, a stereoisomer, a derivative, or a pharmaceutically thereof Acceptable salts. The compound according to claim 1, wherein the compound provides the activity of inhibiting cell migration, inhibiting cancer cell migration, or inhibiting cancer metastasis, a stereoisomer, a derivative, or a pharmaceutically acceptable salt thereof. A pharmaceutical composition for preventing or treating cancer, comprising the compound according to any one of claims 1 to 4, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof. The method according to claim 5, wherein the cancer is lung cancer, laryngeal cancer, stomach cancer, colon/rectal cancer, liver cancer, gallbladder cancer, pancreatic cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, prostate cancer, carcinoma derived from epithelial cells such as skin cancer, etc. ( Carcinoma), bone cancer, muscle cancer, adipose cancer, sarcoma derived from connective tissue cells such as fibroblast cancer, hematopoietic cancer derived from hematopoietic cells such as leukemia, lymphoma, multiple myeloma, tumors occurring in nerve tissue, Triple negative breast cancer, brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroglioma, intracranial tumor, ependymoma, brainstem tumor, head and neck tumor, laryngeal cancer, oropharyngeal cancer, nasal/ Sinus cancer, nasopharyngeal cancer, salivary gland cancer, hypopharyngeal cancer, thyroid cancer, chest tumor, small cell lung cancer, non-small cell lung cancer, thymus cancer, mediastinum tumor, and a pharmaceutical composition that is at least one selected from the group consisting of esophageal cancer. The pharmaceutical composition according to claim 5, wherein the pharmaceutical composition provides cancer metastasis inhibition or epithelial-mesenchymal cell metastasis (EMT) inhibitory activity. The pharmaceutical composition according to claim 5, further comprising a pharmaceutical additive selected from the group consisting of a diluent, a binder, a disintegrant, a lubricant, and any combination thereof. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is for oral administration. A method for treating cancer, comprising administering the compound according to any one of claims 1 to 4, a stereoisomer, a derivative thereof, or a pharmaceutically acceptable salt thereof to a subject other than a human.

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