Aurora kinase inhibitors and uses thereof

Abstract

The present invention relates to a novel pyridine compound, its preparation method and application. Specifically, the present invention relates to the compound of formula (1) and its preparation method, and the application of the compound of formula (1) and its pharmaceutically acceptable salt as an Aurora kinase inhibitor in the preparation of antitumor drugs. [Formula 1] TIFF2022541467000069.tif35168

Description

[Technical Field]

[0001] The present invention claims the benefit of Chinese Patent Application No. 201910643061.2, filed on July 16, 2019, which is incorporated herein by reference in its entirety.

[0002] FIELD OF THE INVENTION The present invention relates to the fields of medicine, medicinal chemistry and pharmacology, and more particularly to a class of Aurora kinase inhibitors, their preparation methods and uses. [Background technology]

[0003] Background of the Invention Aurora kinases are serine / threonine kinases that play essential roles in centrosome duplication, spindle assembly, chromosome segregation, and the spindle assembly checkpoint during mitosis (Cancer Metastasis Rev., 2003, 22, 451). There are three structurally and functionally related Aurora kinases: Aurora-A, Aurora-B, and Aurora-C. Aurora-A localizes next to the centrosome early in mitosis and associates with spindle microtubules and mitotic poles throughout metaphase and telophase. It is required for centrosome duplication, maturation, and separation, and for bipolar spindle formation, regulating mitotic entry and exit (Nat. Rev. Cancer, 2005, 5, 42). Aurora-B localizes to the centrosome around chromatin early in mitosis and to the mitotic spindle during anaphase. It is central to centrosome function, chromosome alignment and segregation, the spindle checkpoint, and cytokinesis (Mol. Cancer Ther., 2009, 8, 2046-2056). The role of Aurora-C in mitosis is less defined. It is expressed at high levels in the testis and may play a specialized role in male animals (Proc Natl Acad Sci USA, 2002, 99(24):15440-15445).

[0004] The gene encoding Aurora-A maps to 20q13.2, a region frequently amplified in cancers including breast, colon, ovarian, and thyroid cancers. Ectopic expression of Aurora-A in normal cells resulted in centrosome amplification, aneuploidy, chromosomal instability, and telomere elongation, characteristics associated with transformed cells (J. Cell Sci., 2007, 120, 2987). Overexpression of Aurora-A or its activating partner, TPX-2, is thought to contribute to chromosomal instability in human cancers. Furthermore, Aurora-A regulates the function of tumor suppressor and proapoptotic proteins such as p53. For example, phosphorylation of p53 at Ser215 and Ser351 by Aurora-A promotes its functional inactivation and degradation, respectively. This region maps to 20q13.2, a region frequently amplified in cancers including breast, colon, ovarian, and thyroid cancers.

[0005] The gene encoding Aurora-B maps to 17p13.1, a chromosomal region frequently deleted or amplified in certain cancers (J. Crin. Pathol., 2007, 60(2):218-221). Elevated expression of Aurora-B mRNA and protein is observed in colon, oral, and non-small cell lung cancers. Aurora-B is a subunit of the chromosomal passenger complex (CPC). Aurora-B regulates mitosis through phosphorylation of INCENP, CENP-A, and survivin. Furthermore, overexpression of Aurora-B has also been shown to enhance ras signaling.

[0006] Aurora kinases are drug targets for anticancer therapeutics due to their oncogenic properties. Aurora kinase inhibitors have been developed as novel anticancer drugs, and among them, LY-3295668, which has a pyridine core, is an Aurora A-specific inhibitor currently in phase I development (WO2016077161). The structure of LY-3295668 is shown below. [ka] [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2016077161 [Non-patent literature]

[0008] [Non-Patent Document 1] Cancer Metastasis Rev.,2003,22,451 [Non-patent document 2] Nat.Rev.Cancer,2005,5,42 [Non-patent document 3] Mol.Cancer Ther.,2009,8,2046-2056 [Non-patent document 4] Proc Natl Acad Sci USA,2002,99(24):15440-15445 [Non-Patent Document 5] J. Cell Sci.,2007,120,2987 [Non-patent document 6] J. Crin. Pathol.,2007,60(2):218-221 Summary of the Invention [Problem to be solved by the invention]

[0009] However, LY-3295668 and other Aurora-A inhibitors have consistently been associated with weak activity, poor oral bioavailability, or moderate in vivo efficacy. Therefore, the development of novel Aurora-A inhibitors with improved in vitro and in vivo activity is warranted. [Means for solving the problem]

[0010] Summary of the Invention The present invention provides novel kinase inhibitors of general formula (1), their optical isomers, crystalline forms and pharmaceutically acceptable salts. [ka] In formula (1), R 1 is an aryl, heteroaryl, [ka] and wherein the aryl and heteroaryl are optionally substituted with 1 to 3 groups selected from halogen, C1-C3 alkyl, C1-C3 alkoxyl, halogen-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxyl; R 2 is H or methyl, R 3 is H or F, W is [ka] and where R a is H, C1-C3 alkyl or C3-C6 cycloalkyl, R b is H, C1-C3 alkyl or C3-C6 cycloalkyl, and W is [ka] and R b is H, C2-C3 alkyl, or C3-C6 cycloalkyl, L is CH2, CO, CD2, CH(Me), C(Me)2, [ka] or W is [ka] and R b is a methyl group, L is CO, CD2, CHMe, C(Me)2, [ka] is.

[0011] In another preferred embodiment, in formula (1), R 1 teeth, [ka] and where X is NH, O or S, and R c and R d are independently H, halogen, C1-C3 alkyl, C1-C3 alkoxyl, halogen-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxyl.

[0012] In another preferred embodiment, in formula (1), R 1 teeth, [ka] is.

[0013] In another preferred embodiment, in formula (1), W is [ka] and W is [ka] If L is CH2, CO, CD2, CH(Me), C(Me)2, [ka] W is [ka] If L is CO, CD2, CHMe, C(Me)2, [ka] is.

[0014] The present inventors have synthesized and carefully studied various novel compounds with Aurora kinase inhibition and have found that for compounds of general formula (1), when the -L group is changed from CH2 to a group of appropriate size such as CD2, and / or W is [ka] In this case, we found that these compounds possess highly potent Aurora-A kinase inhibitory activity, while also exhibiting significantly improved Aurora-B activity and in vivo antitumor activity.

[0015] In another preferred embodiment, the compound of general formula (1) is selected from the compounds listed in Table 1.

[0016] [Table 1-1] [Table 1-2] [Table 1-3]

[0017] In another embodiment, the present invention provides a combined pharmaceutical composition comprising a pharmacologically acceptable excipient or carrier and the compound of formula (1) of the present invention, its optical isomer, or a pharmaceutically acceptable salt thereof as an active ingredient.

[0018] In another embodiment, the present invention provides the use of a compound, its optical isomer, or a pharmaceutically acceptable salt in the manufacture of a medicament for treating aurora kinase-associated diseases, particularly for application in anti-tumor drugs. DETAILED DESCRIPTION OF THE INVENTION

[0019] Detailed Description of the Embodiments Methods for preparing the compound of general formula (1) are specifically described below, but these specific methods do not limit the present invention.

[0020] The compounds of formula (1) above can be synthesized using standard synthetic techniques, well-known techniques, or a combination of the methods described herein. Additionally, the solvents, temperatures, and other conditions described herein can be used to synthesize the compounds. The reaction conditions for (1) may vary. Starting materials for the synthesis of compounds of formula (1) may be synthesized or obtained from commercial sources such as, but not limited to, Aldrich Chemical Co. (Milwaukee, WI) or Sigma Chemical Co. (St. Louis, MO). The compounds described herein and other related compounds with various substituents are described in March, ADVANCED ORGANIC CHEMISTRY 4th Ed. (Wiley 1992), Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4th Ed., Vols. A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3 rd Ed., (Wiley 1999). The general methods for preparing the compounds can be varied by using appropriate reagents and conditions to introduce various groups into the molecular formulas provided herein.

[0021] In one aspect, the compounds described herein can be obtained according to known methods. However, the process conditions, such as reactants, solvents, bases, the amount of compounds used, reaction temperature, and reaction time, are not limited to the following description. The compounds of the present invention can also be conveniently prepared by any combination of various synthetic methods described herein or known methods, and such combinations can be easily carried out by those skilled in the art. In another aspect, the present invention also provides a method for preparing a compound represented by general formula (1), which is prepared by the following method A or method B.

[0022] Method A includes the following steps: first, starting materials A and B are coupled under basic conditions using a palladium catalyst and a ligand to produce compound C; then, compound C is deprotected by deprotecting the Boc group under acidic conditions to give compound D; and then compound D is reacted with R 1 -LX to give compound E, which is finally hydrolyzed under acidic or basic conditions to give the compound of formula (1a).

[0023] [ka]

[0024] In the above reaction, R 1 , R 2 , R 3 , R a and L have the same definition as defined above, and X is selected from Br, Cl, OTf or OH.

[0025] Method B includes the following steps: first, starting material F is reacted with B to produce compound G, then compounds G and H are coupled under basic conditions using a palladium catalyst and a ligand to produce compound I, and finally compound I is hydrolyzed under strongly acidic conditions to give the compound of formula (1b).

[0026] [ka]

[0027] In the above reaction, R 1 , R 2 , R 3 , R a and L have the same definition as defined above, and X is selected from Br, Cl, OTf or OH.

[0028] Further forms of the compound The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to a subject to which it is administered and does not eliminate the biological activity and properties of the compound. The salt of the compound of the present invention refers to a salt conventionally used in the field of organic chemistry, for example, the compound may be a base addition salt having a carboxyl group, or the compound may be an acid addition salt having an amine group or a basic heterocyclic group.

[0029] Examples of base addition salts include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, and ammonium salts such as trimethylamine salts, triethylamine salts, dicyclohexylamine salts, ethanolamine salts, diethanolamine salts, triethanolamine salts, procaine salts, and N,N'-dibenzylethylenediamine salts.

[0030] Examples of acid addition salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, and phosphate; organic acid salts such as acetate, formate, maleate, fumarate, citrate, oxalate, and ascorbate; and sulfonates such as methanesulfonate, benzenesulfonate, and p-toluenesulfonate.

[0031] It should be understood that pharmaceutically acceptable salts include solvent addition forms or crystalline forms, particularly solvates or polymorphs. Solvates include stoichiometric or non-stoichiometric solvents and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water and ethanol. Hydrates are formed when the solvent is water, or alcoholates are formed when the solvent is ethanol. Solvates of the compound of formula (1) can be conveniently prepared or formed according to the methods described herein. For example, hydrates of the compound of formula (1) are conveniently prepared by recrystallization from a water / organic solvent mixture, where the organic solvent used includes, but is not limited to, dioxane, tetrahydrofuran, ethanol, or methanol. Furthermore, the compounds referred to herein can exist in unsolvated or solvated forms. In summary, for purposes of the compounds and methods provided herein, solvated forms are considered equivalent to unsolvated forms.

[0032] In other specific embodiments, the compound of formula (1) is prepared in various forms, including, but not limited to, amorphous, crushed, and nanoparticle-sized forms. Furthermore, the compound of formula (1) includes crystalline and polymorphic forms. Polymorphic forms include different lattice arrangements of the same elemental composition of the compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardness, crystalline forms, optical and electrical properties, stability, and solubility. Various factors, such as recrystallization solvents, crystallization rates, and storage temperatures, can primarily cause the formation of specific crystals.

[0033] In other embodiments, the compounds of formula (1) have one or more stereocenters and therefore occur in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, and single diastereomers. The possible asymmetric centers depend on the properties of the various substituents on the molecule. Each such asymmetric center independently generates two optical isomers, and all possible optical isomers and diastereomeric mixtures, as well as pure or partially pure compounds, are within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0034] therapeutic use The compounds or compositions described herein generally can be used to inhibit Aurora kinases and, therefore, can be used to treat one or more diseases associated with Aurora kinases. Thus, in certain embodiments, the present invention provides a method for treating an Aurora kinase-mediated disease, comprising administering a compound of the present invention, or a pharmaceutically acceptable composition thereof, to a patient in need thereof.

[0035] Cancers that can be treated with the compounds of the invention include, but are not limited to, hematological malignancies (leukemia, lymphoma, myeloma including multiple myeloma, myelodysplastic syndrome or myelodysplastic syndrome) and solid tumors (such as prostate, breast, lung, colon, pancreatic, kidney, ovarian, soft tissue cancer and osteosarcoma or stromal tumors).

[0036] Administration route The compound of the present invention and its pharmaceutically acceptable salt can be prepared into various preparations, which contain the compound of the present invention or its pharmaceutically acceptable salt and pharmaceutically acceptable excipient or carrier in a safe and effective amount range.In these, "safe and effective amount" means that the amount of compound can clearly improve the condition without causing serious side effects.The safe and effective amount of compound is determined according to the age, disease, treatment course and other specific conditions of the subject.

[0037] "Pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and low toxicity. "Compatible" here means that the components of the composition can be mixed with and among the compounds of the present invention without significantly reducing the effectiveness of the compound. Examples of pharmaceutically acceptable excipients or carriers include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, sodium cellulose acetate, etc.), gelatin, talc, solid lubricants (e.g., stearic acid and magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween), wetting agents (e.g., sodium dodecyl sulfate, etc.), colorants, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0038] The compounds of the present invention may be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously) or topically.

[0039] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with (a) fillers or compatibilizers, such as starch, lactose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, such as glycerol; (d) disintegrating agents, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption enhancers, such as quaternary amine compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also include buffers.

[0040] Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared using coating and shell materials, such as casings and other materials well known in the art.They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a part of the digestive tract.The examples of embedding components that can be used are polymeric substances and waxes.If desired, active compound can also be formed into microcapsules with one or more of the above-mentioned excipients.

[0041] The liquid dosage form for oral administration comprises pharmaceutically acceptable emulsion, solution, suspension, syrup or tincture.In addition to active compound, liquid dosage form can comprise water or other solvent, solubilizer and emulsifier, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide and oil, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil or their mixture, and other inert diluents that are conventionally used in the art.

[0042] Besides these inert diluents, compositions can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening agents, flavoring agents, and flavorants.

[0043] In addition to the active compound, suspensions may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide, agar, or mixtures thereof.

[0044] Compositions for parenteral injection can include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0045] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0046] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0047] When a pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is administered to a mammal (e.g., a human) in need of treatment, wherein the dosage upon administration is a pharmaceutically acceptable effective amount, and for a human weighing 60 kg, the daily dose is generally 1 to 1000 mg, preferably 10 to 500 mg. Of course, the specific dose should also take into account factors such as the route of administration and the patient's health, which are within the skill of a skilled physician.

[0048] It should be noted that the features described above in the present invention or the features described above in the embodiments can be randomly combined. All features disclosed herein may be used in any configuration, and various features disclosed herein may be replaced with any alternative features that serve the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0049] Various specific aspects, features and advantages of the above compounds, methods and pharmaceutical compositions are described in detail as follows. It should be understood that the following detailed description and examples describe specific embodiments for reference only. After reading the description of the present invention, various changes or modifications may occur to those skilled in the art, and such equivalents fall within the scope of this application.

[0050] In all examples, 1 H-NMR was recorded on a Varian Mercury 400 NMR spectrometer, and chemical shifts were expressed as δ (ppm). Silica gel for separation was 200-300 mesh unless otherwise specified, and the ratio of eluents was by volume.

[0051] The abbreviations used in this invention are as follows: ACN represents acetonitrile, Ar represents argon, CBr4 represents carbon tetrabromide, CDCl3 represents deuterated chloroform, CD3OD represents deuterated methanol, DCM represents dichloromethane, DIPEA represents diisopropylethylamine, Diox represents 1,4-dioxane, DMF represents dimethylformamide, EA represents ethyl acetate, EDCl represents 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, h represents hours, HOBt represents 1-hydroxybenzotriazole, K2CO3 represents potassium carbonate, KI represents potassium iodide, K3PO 4 represents potassium phosphate, LC-MS represents liquid-mass spectroscopy, LiAlD4 represents lithium aluminum deuteride, LiOH represents lithium hydroxide, mL represents milliliter, MeOH represents methanol, min represents minute, MS represents mass spectroscopy, NMR represents nuclear magnetic resonance, Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium, PE represents petroleum ether, PPh3 represents triphenylphosphine, Tf2O represents trifluoromethanesulfonic anhydride, and Xantphos represents 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene. [Example]

[0052] Detailed Description of the Invention

[0053] Example 1: Synthesis of 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 1) [ka]

[0054] 1-(tert-butyl)4-methyl4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylic acid 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate (5 g, 11.23 mmol, synthesized by referring to the method of WO2016 / 077161), thiazol-2-amine (956 mg, 9.55 mmol), anhydrous potassium phosphate (6 g, 28.08 mmol), Xantphos (650 mg, 1.123 mmol), and dioxane (100 mL) were added to a 250 mL flask, and Pd2(dba)3 (514 mg, 0.562 mmol) was added after Ar replacement, and the mixture was heated to reflux temperature and reacted for another 5 hours under Ar protection. After the reaction was determined to be complete by LC-MS, it was concentrated under reduced pressure and further purified by column chromatography (DCM / MeOH=50 / 0 to 50 / 1) to give the desired product as a yellow solid (4.0 g, 89% yield), ESI-MS m / z: 465.2 [M+H] + was obtained as.

[0055] Methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate 1-(tert-butyl) 4-methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate (4 g, 8.61 mmol) was added to a 100 mL flask, and DCM (20 mL) and HCl / dioxane (22 mL, 4 M, 88 mmol) were also added, followed by stirring at room temperature for 20 h. After the reaction was determined to be complete by LC-MS, it was concentrated, EA (30 mL) was added to the residue, stirred for an additional 30 min, filtered, and dried to give a yellow solid (4.1 g, 100% yield), ESI-MS m / z: 365.2 [M+H]. + The target compound was obtained as

[0056] Methyl 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate Methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (800 mg, 1.83 mmol), 1-(bromomethyl)-3-chloro-2-fluorobenzene (500 mg, 2.19 mmol), K2CO3 (1.264 g, 9.15 mmol), KI (20 mg), and ACN (20 mL) were added to a 100 mL flask, and the mixture was reacted at room temperature for 2 hours. After the reaction was determined to be complete by LC-MS, water (100 mL) was added to precipitate a solid, which was then filtered. The filter cake was washed twice with water (20 mL × 2) and then slurried with PE (50 mL). After filtration, the filter cake was washed twice with PE (20 mL x 2) and dried to give the crude product, methyl 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (935 mg, 100% yield), which was carried on to the next step without further purification. ESI-MS m / z: 510.2 [M+H] + .

[0057] 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid Methyl 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (935 mg, 1.83 mmol) was added to a 100 mL flask, followed by water (15 mL) and concentrated HCl (15 mL), heated to reflux, and reacted for an additional 5 hours. After the reaction was determined to be complete by LC-MS, it was concentrated, and the residue was slurried with ACN (30 mL) at room temperature. After filtration, the filter cake was washed with ACN (5 mL × 2) and dried to give the desired yellow powder (818 mg, 90% yield).

[0058] 1H NMR (400MHz, DMSO-d6)δ:11.68(s, 1H), 10.59(s, 1H), 7.76(t, J=7.2Hz, 1H), 7.73-7.6 8(m, 1H), 7.61(t, J=9.2Hz, 1H), 7.43(d, J=3.7Hz, 1H), 7.34(t, J=7.9Hz, 1H), 7.01(q, J=3.9Hz, 2H), 4.72(d, J=13.3Hz, 1H), 4.36(dd, J=13.6, 8.3Hz, 1H), 3.88(s, 2H), 3.26 -3.21(m, 2H), 3.09(d, J=12.9Hz, 1H), 2.16-1.95(m, 4H), 1.50(d, J=6.0Hz, 3H);ESI-MS m / z:493.1[m+H] + .

[0059] Four different optical isomers of compound 1 can be obtained by synthesis from different chiral starting materials or by separation by chiral SFC, and the structures are as follows: [ka]

[0060] Compounds 1-1, 1-2, 1-3 and 1-4 are designated as follows: 1-1: (2R,4R)-1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid, 1-2: (2S,4S)-1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid, 1-3: (2R,4S)-1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid, 1-4: (2S,4R)-1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid.

[0061] Other compounds in this application can also be used to separate the corresponding optical isomers by the same method.

[0062] Example 2: Synthesis of 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-1-((3-fluoropyridin-4-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 2) [ka]

[0063] The target compound was obtained by the same synthesis method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)-3-fluoropyridine as starting materials.

[0064] 1 H NMR (400MHz, DMSO-d6)δ:12.45(s, 1H), 11.45(s, 1H), 8.74(d, J=1.4Hz, 1H), 8.54(d, J=4.9Hz, 1H), 8.0 3(t, J=5.7Hz, 1H), 7.68(t, J=9.1Hz, 1H), 7.50(d, J=4.0Hz, 1H), 7.13(q, J=3.7, 3.3Hz, 2H), 4.71(d, J= 13.3Hz, 1H), 4.43(dd, J=13.5, 7.9Hz, 1H), 3.93(s, 1H), 3.39(dt, J=14.8, 9.1Hz, 1H), 3.30-3.23(m, 2H) ), 3.09(d, J=13.1Hz, 1H), 2.08(t, J=15.2Hz, 3H), 1.92(d, J=15.2Hz, 1H), 1.51(d, J=6.2Hz, 3H); ESI-MS m / z:460.2[M+H] + .

[0065] Example 3: Synthesis of 1-((2-chloro-3-fluoropyridin-4-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 3) [ka]

[0066] The target compound was obtained by the same synthesis method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)-2-chloro-3-fluoropyridine as starting materials.

[0067] 1 H NMR (400MHz, DMSO-d6)δ:11.22(s, 1H), 9.82(s, 1H), 8.39(s, 1H), 7.68(s, 1H), 7.57(t, J=9.1Hz, 1H), 7.37(d, J=3.6Hz, 1H), 6 ESI-MS m / z:494.1[M+H] + .

[0068] Example 4: Synthesis of 1-((3-fluoro-2-(2,2,2-trifluoroethoxy)pyridin-4-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 4) [ka]

[0069] The target compound was obtained by the same synthesis method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)-3-fluoro-2-(2,2,2-trifluoroethoxy)pyridine as starting materials.

[0070] 1 H NMR (400MHz, methanol-d4) δ:7.64(dd, 8.3, 7.4Hz, 1H), 7.32-7.7(m, 2H), 7.23(t, 7.9Hz, 1H), 7.17(dd, 7.8, 1.5Hz, 1H), 7.02(d, 7.2Hz, 1H), 6 .906.86(m, 2H), 3.84-3.77(m, 2H), 3.73(s, 2H), 3.44(t, J=5.1Hz, 2H), 2.69(t, 5.1Hz, 2H), 2.64-2.58(m, 3H), 1.53(d, J=6.0Hz, 3H);[M+H] + .

[0071] Example 5: Synthesis of 1-((5-chlorothiophen-2-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 5) [ka]

[0072] The target compound was obtained by the same synthesis method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 2-(bromomethyl)-5-chlorothiophene as starting materials.

[0073] 1H NMR (400MHz, methanol-d4) δ:7.69(t, J=8.7Hz, 1H), 7.58(s, 1H), 7.25(q, 9.5, 7.5Hz, 2H), 7.15(dd, 9.0, 3.0Hz, 1H), 7.00(dd, 30.0, 3.5H) z, 1H), 4.62-4.37(m, 2H), 3.80(d, 26.4Hz, 2H), 3.39(d, J=6.4Hz, 2H), 3.21-3.11(m, 1H), 2.33-1.98(m, 4H), 1.45(d, J=6.0Hz)ESI-MS m / z:481.1[M+H + ].

[0074] Example 6: Synthesis of 1-(benzofuran-4-ylmethyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 6) [ka] The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)benzofuran as starting materials.

[0075] 1 H NMR (400MHz, DMSO-d6)δ:7.62(d, J=7.2Hz, 1H), 7.50(t, J=9.1Hz, 1H), 7.32(d, J=3.4Hz, 1H), 7.24 (dd, J=7.1, 2.1Hz, 1H), 7.19-7.11(m, 2H), 6.89(d, J=5.1Hz, 2H), 6.65(d, J=7.2Hz, 1), 3.84(d, J=1 4.1Hz, 1H), 3.69(d, J=14.2Hz, 1H), 3.05(s, 2H), 2.77-2.73(m, 1H), 2.65-2.57(m, 1H), 2.492.42( m, 1H), 1.84-1.75(m, 1H), 1.72-1.63(m, 2H), 1.57(t, J=11.9Hz, 1H), 1.16(d, J=6.0Hz, 3H); ESI-MS m / z:481.2[M+H] + ..

[0076] Example 7: Synthesis of 1-((2,2-difluorobenzo[d][1,3]dioxol-4-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 7) [ka]

[0077] Methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)-2,2-difluorobenzo[d][1,3]dioxole were used as starting materials to give the intermediate methyl 1-((2,2-difluorobenzo[d][1,3]dioxol-4-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate.

[0078] The above intermediate was added to a 100 mL flask, followed by THF (10 mL), HO (5 mL), and LiOH HO (79 mg, 1.88 mmol), and the mixture was heated to 60 °C and stirred under argon for 5 h. After completion of the reaction as determined by LC-MS, the mixture was concentrated to a volume of approximately 7.5 mL, and the residue was purified by reverse-phase flash to give the target compound (60 mg, 62% yield).

[0079] 1H NMR (400MHz, DMSO-d6) δ:7.50(t, J=9.1Hz, 1H), 7.32(d, J=3.4Hz, 1H), 7.24(dd, J=7.1, 2.1Hz, 1H), 7.19-7.11(m, 2H), 6.89(d, J=5.1Hz, 2H), 3.94(d, J=14.1Hz, 1H), 3.39(d, J= 14.2Hz, 1H), 3.05(s, 2H), 2.75-2.71(m, 1H), 2.61-2.52(m, 1H), 2.45-2.40(m, 1H), 1.8 3-1.79(m, 1H), 1.75-1.62(m, 2H), 1.54(t, J=11.9Hz, 1H), 1.06(d, J=6.0Hz, 3H); ESI-MS m / z:521.2[M+H] + .

[0080] Example 8: Synthesis of 1-((2,2-difluoro-2,3-dihydro-1H-inden-4-yl)methyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 8) [ka]

[0081] The target compound was obtained by the same synthesis method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 4-(bromomethyl)-2,2-difluoro-2,3-dihydro-1H-indene as starting materials.

[0082] 1H NMR (400MHz, DMSO-d6) δ:7.52(t, J=9.1Hz, 1H), 7.30(d, J=3.4Hz, 1H), 7.23(dd, J=7.1, 2.1H z, 1H), 7.19-7.11(m, 2H), 6.89(d, J=5.0Hz, 2H), 5.34-5.21(m, 4H), 3.94(d, J=14.1Hz, 1H), 3 .39(d, J=14.2Hz, 1H), 3.05(s, 2H), 2.75-2.71(m, 1H), 2.61-2.50(m, 1H), 2.45-2.40(m, 1H) , 1.83-1.79(m, 1H), 1.75-1.65(m, 2H), 1.54(t, J=11.9Hz, 1H), 1.07(d, J=6.0Hz, 3H);ESI-MS m / z:517.2[M+H] + .

[0083] Example 9: Synthesis of 1-(1-(3-chloro-2-fluorophenyl)ethyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 9) [ka]

[0084] The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 1-(1-bromoethyl)-3-chloro-2-fluorobenzene as starting materials.

[0085] 1H NMR (400MHz, DMSO-d6)δ:12.27(s, 1H), 11.15(s, 1H), 7.55(d, J=8.1Hz, 1H), 7. 49-7.34(m, 2H), 7.18(t, J=7.8Hz, 1H), 7.02(s, 1H), 6.86(d, J=8.3Hz, 1H), 6.6 7(d, J=7.3Hz, 1H), 4.41(m, 1H), 3.11(d, J=16.6Hz, 2H), 3.02(m, 2H), 2.85(m, 1 H), 1.76-1.52(m, 4H), 1.30(d, J=6.7Hz, 3H), 1.06(q, J=7.1, 6.4Hz, 3H); ESI-MS m / z:507.2[M+H] + .

[0086] Example 10: Synthesis of 1-(1-(3-chloro-2-fluorophenyl)cyclopropyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 10) [ka]

[0087] Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-(1-(3-chloro-2-fluorophenyl)-cyclopropyl)-2-methylpiperidine-4-carboxylate 1-(3-Chloro-2-fluorophenyl)cyclopropan-1-ol (400 mg, 2.145 mmol) was added to a 100 mL flask, followed by dry DCM (10 mL) and DIPEA (692 mg, 5.362 mmol). The mixture was cooled to -45 °C under argon protection, and then TfO (726 mg, 2.574 mmol) in 10 mL of DCM was added and stirred at -50 to -40 °C for 2 h. Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (444 mg, 1.287 mmol) in 10 mL of CHCN was added, and the mixture was allowed to warm to room temperature and stirred for 2 h. After the reaction was determined to be complete by LC-MS, it was quenched with water (20 mL), the organic phase was separated and extracted with DCM (20 mL), the organic phases were combined and concentrated, and the residue was purified by column chromatography to give the target compound (284 mg, 43% yield), ESI-MS m / z: 533.2 [M+H] + obtained.

[0088] 1-(1-(3-chloro-2-fluorophenyl)cyclopropyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-(1-(3-chloro-2-fluorophenyl)cyclopropyl)-2-methylpiperidine-4-carboxylate as a starting material.

[0089] 1H NMR (400MHz, DMSO-d6)δ:12.11(s, 1H), 11.02(s, 1H), 7.57(d, J=8.1Hz, 1H), 7.43-7.33(m, 2H), 7.15(t, J=7.8Hz, 1H), 7.02(s, 1H), 6.84(d, J ESI-MS m / z:519.2[M+H] + .

[0090] Example 11: Synthesis of 1-(3-(3-chloro-2-fluorophenyl)oxetan-3-yl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 11) [ka]

[0091] Using methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate and 3-(3-chloro-2-fluorophenyl)oxetan-3-ol as starting materials, the intermediate methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-(3-(3-chloro-2-fluorophenyl)oxetan-3-yl)-2-methylpiperidine-4-carboxylate was obtained by the same synthetic method as in Example 10.

[0092] The target compound was obtained by the same synthesis method as in Example 1 using the above intermediate and thiazol-2-amine as starting materials.

[0093] 1H NMR (400MHz, CD3OD) δ:7.46(m, 2H), 7.35-7.16(m, 3H), 6.93(dt, J=8.9, 3.3Hz, 1H), 6.8 7(t, J=4.1Hz, 1H), 4.77(s, 1H), 4.70-4.47(m, 2H), 4.31(d, J=12.2Hz, 1H), 4.14(d, J=8. 0Hz, 1H), 3.98(ab, J=29.9, 12.4Hz, 1H), 3.67(t, J=11.3Hz, 1H), 3.17-2.96(m, 2H), 2.5 0-2.13(m, 3H), 2.08-1.95(m, 1H), 1.89(d, J=12.4Hz, 1H), 1.41(d, J=6.9Hz, 3H); ESI-MS m / z:535.2[M+H] + .

[0094] Example 12: Synthesis of 1-(3-chloro-2-fluorobenzoyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 12) [ka]

[0095] 3-Chloro-2-fluorobenzoic acid (262 mg, 1.50 mmol), DMF (20 mL), EDCI (431 mg, 2.25 mmol), HOBt (304 mg, 2.25 mmol), and DIPEA (970 mg, 7.52 mmol) were added to a 100 mL flask, and the mixture was stirred at room temperature under Ar protection for 30 minutes. Methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride (437 mg, 1.0 mmol) was added and stirred at room temperature for 20 hours. After the reaction was determined to be complete by LC-MS, it was quenched with water (40 mL) and extracted with EA (50 mL × 2). The organic phases were combined, washed with saturated NaCl solution, concentrated, and the residue was purified by column chromatography to give the desired intermediate (365 mg, 70% yield).

[0096] The target compound was obtained by the same synthesis method as in Example 1 using the above intermediate as a starting material.

[0097] 1 H NMR (400MHz, CD3OD) δ:7.68(t, J=9.0Hz, 1H), 7.62-7.52(m, 2H), 7.43(dt, J=12.0, 7.9Hz, 1H), 7.27(d, J=7.5Hz, 1H), 7.21(d ESI-MS m / z:475.2[M+H] + .

[0098] Example 13: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 13) [ka]

[0099] 1-(Bromomethyl-d2)-3-chloro-2-fluorobenzene Methyl 3-chloro-2-fluorobenzoate (658 mg, 3.49 mmol) and THF (dry, 10 mL) were added to a 100 mL flask, and LiAlD (146 mg, 3.49 mmol) was added dropwise in an ice bath and stirred for another 0.5 h. After completion of the reaction based on TLC (PE / EA = 10 / 1), the reaction was quenched with water (20 mL) in an ice bath, saturated NaCl solution (20 mL) was added, and the mixture was extracted with EA (20 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to give the crude intermediate (3-chloro-2-fluorophenyl)methane-d2-ol, which was used in the next step without further purification.

[0100] (3-Chloro-2-fluorophenyl)methan-d2-ol, DCM (20 mL), and CBr4 (2 g, 6.03 mmol) were added to a 100 mL flask, and PPh3 (1.371 g, 5.23 mmol) was added dropwise and stirred at room temperature for approximately 1 h. After completion of the reaction determined by TLC (PE / EA = 10 / 1), the mixture was concentrated, and the residue was purified by column chromatography (PE, 800 mL) to give 1-(bromomethyl-d2)-3-chloro-2-fluorobenzene as a colorless liquid (921 mg, 100% yield).

[0101] 1 H NMR (400MHz, CDCl3) δ7.34 (ddd, J=8.3, 6.9, 1.7Hz, 1H), 7.27 (ddd, J=7.9, 6.5, 1.7Hz, 1H), 7.05 (td, J=7.9, 1.2Hz, 1H).

[0102] Methyl 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate Methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride (800 mg, 1.83 mmol), 1-(bromomethyl-d)-3-chloro-2-fluorobenzene (495 mg, 2.19 mmol), K2CO3 (1.264 g, 9.15 mmol), KI (20 mg), and ACN (20 mL) were added to a 100 mL flask and stirred at room temperature for approximately 2 hours. After the reaction was determined to be complete by TLC, water (100 mL) was added to precipitate a solid, which was then filtered. The filter cake was washed twice with water (20 mL x 2) and then slurried with PE (50 mL). After filtration, the filter cake was washed with PE (20 mL x 2) and dried to give the crude product, methyl 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (931 mg, 100% yield), which was used in the next step without further purification. ESI-MS m / z: 509.2 [M+H] + .

[0103] 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid Methyl 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (931 mg, 1.83 mmol) was added to a 100 mL flask, followed by water (15 mL) and concentrated HCl (15 mL). The mixture was heated to 105° C. under reflux and allowed to react for an additional 5 hours. After the reaction was determined to be complete by LC-MS, it was concentrated, and the residue was slurried with ACN (30 mL) at room temperature. After filtration, the filter cake was washed with ACN (5 mL×2) and dried to give the desired target compound as a yellow powder (815 mg, 90% yield).

[0104] 1H NMR (400MHz, CD3OD) δ:7.73(t, J=8.7Hz, 1H), 7.70-7.55(m, 3H), 7.36-7.24(m, 2H), 7.20(dd, J=8. 8, 2.9Hz, 1H), 3.96(s, 2H), 3.57-3.44(m, 2H), 3.36(s, 1H), 2.41-1.96(m, 4H), 1.53(m, 3H), ESI-MS m / z:495.1[M+H] + .

[0105] Four different optical isomers of compound 13 can be obtained by synthesis from different chiral starting materials or by separation by chiral SFC, and the structures are as follows: [ka]

[0106] Compounds 13-1, 13-2, 13-3 and 13-4 are designated as follows: 13-1: (2R,4R)-1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid; 13-2: (2S,4S)-1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid; 13-3: (2R,4S)-1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid; 13-4: (2S,4R)-1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid.

[0107] Example 14: Synthesis of 1-(3-chloro-2-fluorobenzyl)-2-methyl-4-((6-(thiazol-2-ylamino)pyridin-2-yl)methyl)piperidine-4-carboxylic acid (Compound 14) [ka]

[0108] The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 1-(bromomethyl)-3-chloro-2-fluorobenzene as starting materials.

[0109] 1 H NMR (400MHz, DMSO-d6)δ:11.88(s, 1H), 10.79(s, 1H), 7.74(t, J=7.2Hz, 1H), 7.72-7.6 3(m, 2H), 7.55(t, J=9.2Hz, 1H), 7.41(d, J=3.7Hz, 1H), 7.32(t, J=7.9Hz, 1H), 7.06(q, J=3.9Hz, 2H), 4.73(d, J=13.3Hz, 1H), 4.38(dd, J=13.6, 8.3Hz, 1H), 3.68(s, 2H), 3.26 -3.24(m, 2H), 3.02(d, J=12.9Hz, 1H), 2.15-1.91(m, 4H), 1.52(d, J=6.0Hz, 3H);ESI-MS m / z:507.1[M+H] + .

[0110] Example 15: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-2-methyl-4-((6-(thiazol-2-ylamino)pyridin-2-yl)methyl)piperidine-4-carboxylic acid (Compound 15) [ka]

[0111] The target compound was obtained by the same synthesis method as in Example 1 using methyl 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate and thiazol-2-amine as starting materials.

[0112] 1 H NMR (400MHz, CD3OD) δ:7.78(t, J=8.7Hz, 1H), 7.70-7.55(m, 3H), 7.36-7.24(m, 3H), 7.23(dd, J=8. 8, 2.9Hz, 1H), 3.92(s, 2H), 3.57-3.44(m, 2H), 3.36(m, 1H), 2.41-1.96(m, 4H), 1.55(m, 3H);ESI-MS m / z:477.1[M+H] + .

[0113] Example 16: Synthesis of 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-((5-methylthiazol-2-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 16) [ka]

[0114] The target compound was obtained by the same synthesis method as in Example 1 using 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate and 5-methylthiazol-2-amine as starting materials.

[0115] 1H NMR (400MHz, DMSO-d6)δ:11.66(s, 1H), 10.59(s, 1H), 7.76(t, J=7.2Hz, 1H), 7.72 -7.66(m, 2H), 7.55(t, J=9.2Hz, 1H), 7.34(t, J=7.9Hz, 1H), 7.01(m, 1H), 4.72(d, J=13.3Hz, 1H), 4.36(dd, J=13.6, 8.3Hz, 1H), 3.88(s, 2H), 3.26-3.21(m, 2H), 3.0 9(d, J=12.9Hz, 1H), 2.27(s, 3H), 2.16-1.95(m, 4H), 1.50(d, J=6.0Hz, 3H);ESI-MS m / z:508.1[M+H] + .

[0116] Example 17: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-((5-methylthiazol-2-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 17) [ka]

[0117] The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 1-(bromomethyl-d2)-3-chloro-2-fluorobenzene as starting materials.

[0118] 1 H NMR (400MHz, CD3OD) δ:7.75(t, J=8.7Hz, 1H), 7.72-7.56(m, 3H), 7.28(s, 1H), 7.20(dd, J=8.8, 2.9Hz, 1H), 3.96(s, 2H), 3.57-3.44(m, 2H), 3.36(s, 1H), 2.57(s, 3H), 2.41-1.96(m, 4H), 1.53(m, 3H), ESI-MS m / z:510.0[M+H] + .

[0119] Example 18: Synthesis of 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-((4-methylthiazol-2-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 18) [ka]

[0120] The target compound was obtained by the same synthesis method as in Example 1 using 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate and 4-methylthiazol-2-amine as starting materials.

[0121] 1 H NMR (400MHz, DMSO-d6)δ:11.64(s, 1H), 10.53(s, 1H), 7.73(t, J=7.2Hz, 1H), 7.70 -7.66(m, 2H), 7.55(t, J=9.2Hz, 1H), 7.34(t, J=7.9Hz, 1H), 6.56(s, 1H), 4.68(d, J=13.3Hz, 1H), 4.37(dd, J=13.6, 8.3Hz, 1H), 3.83(s, 2H), 3.26-3.21(m, 2H), 3.0 6(d, J=12.9Hz, 1H), 2.17(s, 3H), 2.14-1.93(m, 4H), 1.51(d, J=6.0Hz, 3H); ESI-MS m / z:508.1[M+H] + .

[0122] Example 19: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-((4-methylthiazol-2-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 19) [ka]

[0123] The target compound was obtained by the same synthetic method as in Example 1 using methyl 4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate hydrochloride and 1-(bromomethyl-d2)-3-chloro-2-fluorobenzene as starting materials.

[0124] 1 H NMR (400MHz, CD3OD) δ:7.76(t, J=8.7Hz, 1H), 7.73-7.57(m, 3H), 7.22(dd, J=8.8, 2.9Hz, 1H), 6.58(s, ESI-MS m / z:510.0[M+H] + .

[0125] Example 20: Synthesis of 1-(3-chloro-2-fluorobenzyl)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2,2-dimethylpiperidine-4-carboxylic acid (Compound 20) [ka]

[0126] The target compound was obtained by the same synthetic method as in Example 1 using 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2,2-dimethylpiperidine-1,4-dicarboxylate and thiazol-2-amine as starting materials.

[0127] 1H NMR (400MHz, CD3OD) δ:7.76(t, J=8.7Hz, 1H), 7.70-7.51(m, 3H), 7.35-7.24(m, 2H), 7.18(dd, J=8.8, 2.9Hz, 1H), 4.61(d, J=13 ESI-MS m / z:508.2[M+H] + .

[0128] Example 21: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-(thiazol-2-ylamino)pyridin-2-yl)methyl)-2,2-dimethylpiperidine-4-carboxylic acid (Compound 21) [ka]

[0129] The target compound was obtained by the same synthetic method as in Example 1 using 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2,2-dimethylpiperidine-1,4-dicarboxylate and thiazol-2-amine as starting materials.

[0130] 1 H NMR (400MHz, CD3OD) δ:7.75(t, J=8.7Hz, 1H), 7.70-7.55(m, 3H), 7.37-7.26(m, 2H), 7.20(dd, J=8. 8, 2.9Hz, 1H), 3.96(s, 2H), 3.57-3.44(m, 2H), 2.32(s, 2H), 2.02-1.85(m, 2H), 1.43(s, 6H);ESI-MS m / z:509.2[M+H] + .

[0131] Example 22: Synthesis of 1-(3-chloro-2-fluorobenzoyl)-4-((3-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Example 22) [ka]

[0132] Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-(3-chloro-2-fluorobenzoyl)-2-methylpiperidine-4-carboxylate 3-Chloro-2-fluorobenzoic acid (262 mg, 1.50 mmol), DMF (20 mL), EDCI (431 mg, 2.25 mmol), HOBt (304 mg, 2.25 mmol), and DIPEA (970 mg, 7.52 mmol) were added to a 100 mL flask. The mixture was stirred at room temperature for 2 minutes under Ar protection. Methyl 4-((3-fluoro-6-(thiazol-2-thiazol-2-ylamino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate (345 mg, 1.0 mmol) was added and stirred at room temperature for 20 hours. After completion of the reaction by LC-MS monitoring, the reaction was quenched with water (40 mL) and extracted with EA (50 mL × 2). The organic phases were combined, washed with saturated NaCl solution, concentrated, and the residue was purified by column chromatography to give the desired intermediate (426 mg, 85% yield). ESI-MS m / z: 501.1 / 503.1 [M+H] + .

[0133] 1-(3-chloro-2-fluorobenzoyl)-4-((3-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-(3-chloro-2-fluorobenzoyl)-2-methylpiperidine-4-carboxylate (426 mg, 0.85 mmol), tert-butyl 3-amino-5-methyl-1H-pyrazole-1-carboxylate (201 mg, 1.02 mmol), Pd(dba) (92 mg, 0.10 mmol), Xantphos (116 mg, 0.20 mmol), KPO (96 mg, 0.45 mmol), and 1,4-dioxane (10 mL) were added to a 100 mL flask, heated to 100 °C, and reacted under Ar protection for approximately 5 hours. After the reaction was determined to be complete by LC-MS, it was concentrated under reduced pressure and further purified by column chromatography (DCM / MeOH = 10 / 0 to 5 / 1) to give the desired product as a yellow foam (394 mg, 75% yield). ESI-MS m / z: 618.1 [M+H] + .

[0134] The target compound was obtained by the same synthesis method as in Example 1 using the above intermediate as a starting material.

[0135] 1 H NMR (400MHz, CD3OD) δ:7.65(dt, J=16.0, 8.4Hz, 3H), 7.35(t, J=7.9Hz, 1H), 6.92(dd, J=9.0, 3.1Hz, 1H), 6.14-6.05 ESI-MS m / z:504.2[M+H] + .

[0136] Example 23: Synthesis of 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Example 23) [ka]

[0137] Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-((3-chloro-2-fluorophenyl)methyl-d2)-2-methylpiperidine-4-carboxylate The synthesis was carried out by adding 1-(tert-butyl) 4-methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-1,4-dicarboxylate (1 g, 2.62 mmol, synthesized by referring to the method of WO2016 / 077161), 1-(bromomethyl-d2)-3-chloro-2-fluorobenzene (650 mg, 2.88 mmol), K2CO3 (1.811 g, 13.1 mmol), KI (10 mg), and ACN (20 mL) to a 100 mL flask and stirring at room temperature for about 2 hours. After the reaction was determined to be complete by LC-MS, it was concentrated and further purified by column chromatography (PE / EA = 20 / 1 to 8 / 1) to give methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-((3-chloro-2-fluorophenyl)-methyl-d2)-2-methylpiperidine-4-carboxylate (985 mg, 77% yield) as a colorless oil. ESI-MS m / z: 489.1 / 491.1 [M+H] + .

[0138] Methyl 4-((6-((1-(tert-butoxycarbonyl)-5-methyl-1H-pyrazol-3-yl)amino)-3-fluoropyridin-2-yl)methyl)-1-((3-chloro-2-fluorophenyl)methyl-d2)-2-methylpiperidine-4-carboxylate Methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-1-((3-chloro-2-fluorophenyl)-methyl-d2)-2-methylpiperidine-4-carboxylate (985 mg, 2.01 mmol), tert-butyl-3-amino-5-methyl-1H-pyrazole-1-carboxylate (476 mg, 2.41 mmol), Pd2(dba)3 (92 mg, 0.10 mmol), Xantphos (116 mg, 0.20 mmol), K3PO4 (1.067 mmol), and 1,4-dioxane (20 mL) were added to a 100 mL flask heated to 100 °C. The reaction was continued for 5 h under Ar protection. After the reaction was determined to be complete by LC-MS, it was concentrated and further purified by column chromatography (PE / EA = 10 / 1 to 5 / 1) to give the desired intermediate (1.03 g, 84% yield) as a yellow foam. ESI-MS m / z: 605.3 [M+H] + .

[0139] 1-((3-chloro-2-fluorophenyl)methyl-d2)-4-((3-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid Methyl 4-(((6-(1-(tert-butoxycarbonyl)-5-methyl-1H-pyrazol-3-yl)amino)-3-fluoropyridin-2-yl)methyl)-1-((3-chloro-2-fluorophenyl)methyl-d2)-2-methylpiperidine-4-carboxylate (1.03 g, 1.70 mmol) was added to a 100 mL flask, followed by water (15 mL) and concentrated HCl (15 mL), and heated to 105 °C and refluxed for 5 hours. After the reaction was determined to be complete by LC-MS, it was concentrated, and the residue was slurried with ACN (30 mL) at room temperature. After filtration, the filter cake was washed with ACN (5 mL × 2) and dried to give the desired yellow powder (844 mg, 88% yield).

[0140] 1H NMR (400MHz, CD3OD) δ:7.63(dt, J=16.0, 8.4Hz, 3H), 7.32(t, J=7.9Hz, 1H), 6.90(dd, J=9.0, 3.1Hz, 1H), 6.04-6.01 ESI-MS m / z:492.2[M+H] + .

[0141] Example 24: Synthesis of 1-(3-chloro-2-fluorobenzyl)-4-((6-((5-cyclopropyl-1H-pyrazol-3-yl)amino)-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylic acid (Compound 24) [ka]

[0142] The target compound was obtained by the same synthetic method as in Example 23 using methyl 4-((6-bromo-3-fluoropyridin-2-yl)methyl)-2-methylpiperidine-4-carboxylate and 1-(bromomethyl)-3-chloro-2-fluorobenzene as starting materials.

[0143] 1 H NMR (400MHz, CD3OD) δ:7.61(dt, J=16.0, 8.4Hz, 3H), 7.34(t, J=7.9Hz, 1H), 6.85(dd, J=9.0, 3.1Hz, 1H), 6.01-5.95(m, 1H), 4.56(d, J=13.3Hz, 1H) ), 4.13(dd, J=13.6, 8.3Hz, 1H), 3.93(dd, J=11.5, 6.1Hz, 1H), 3.68-3.41(m, 4H), 2.23-2.00(m, 4H), 1.61-1.48(m, 4H), 0.85-0.67(m, 4H); ESI-MS m / z:516.2[M+H] + .

[0144] Example 25: Aurora kinase activity assay The Caliper mobility shift assay was used to measure compound activity against Aurora kinase. Ten different concentrations of compounds were prepared by 3-fold serial dilution. Recombinant Aurora kinase in kinase buffer (20 mM HEPES, pH 7.5, 0.01% Triton X-100) was incubated with the compounds at room temperature for 10 minutes. Subsequently, FAM-labeled peptide substrate was added and the reaction was initiated at 25°C. After the reaction was terminated, the conversion rate was measured by caliper. LY-3295668 was used as a positive control. Data were normalized to that of the vehicle control, and the inhibition rate and IC 50 The results are shown in Table 2.

[0145] Example 26: H1975 proliferation assay H1975 cells growing in logarithmic phase were trypsinized and triturated into a single-cell suspension, and 5 × 10 cells were collected in 50 ml. 3 Cells were seeded into 384-well plates at a concentration of 1000 cells / well. Cells were allowed to attach overnight, and compounds were added to the cells for a further 72 hours of incubation. 50 mL of CTLs were then added, and cell viability was measured by ATP quantification. IC 50 was calculated using GRAPHPAD and is shown in Table 2.

[0146] Table 2: IC for Aurora kinase inhibition and anti-proliferation of H1975 50 value

[0147] [Table 2]

[0148] As listed above, compounds of general formula (1) may be of suitable size where L is CD2 instead of CH2, and / or W is [ka] and show improved efficacy against Aurora-A, Aurora-B, and H1975. The optical isomers in the present disclosure exhibit different activity than the racemic mixture. For example, the optical isomer Compound 1-1 is more potent than its racemic mixture, Compound 1. The activity of other optical isomers of the present disclosure can be similarly measured and may exceed the activity of Compound 1-1.

[0149] Example 27: In vivo anti-tumor growth in H1975 mouse xenografts H1975 cells were cultured in 1640 medium containing 10% FBS at 37°C and 5% CO2. The cells were passaged and harvested by trypsinization. 8 × 10 cells were injected into the left axilla of nude mice. 6 Mice were transplanted with 1000 cells until the tumor volume reached approximately 80 mm 3 When the mice reached 100 mg / kg, they were randomized into four groups of six mice each and administered vehicle, 6 mg / ml LY-3295668, Compound 13, and Compound 23 at 0.1 mg / 10 g, respectively, by oral gavage. Tumor volume and body weight were monitored every other day. Mice were sacrificed on the 21st day of treatment. Relative tumor volume (RTV) and tumor growth (T / C), as well as tumor growth inhibition (TGI), were calculated and analyzed. The results are shown in Table 3.

[0150] Table 3: In vivo efficacy in a mouse H1975 xenograft model

[0151] [Table 3]

[0152] *: P<0.05 vs. control, **: P<0.01 vs. control, ***: P<0.001 vs. control, ****: P<0.0001 vs. control; d1: first day of treatment; d21: last day of treatment; qd*21: once daily for 21 days; RTV: relative tumor volume; RTV=V t / V0; t / C(%)=t RTV / C RTV ×100; t RTV : relative tumor volume (RTV) of the treatment group; C RTV: Relative tumor volume (RTV) of the vehicle control group. TGI: tumor growth inhibition (%); T / C (%)>60%: no effect; T / C (%)≦60% and P<0.05: effective.

[0153] As shown in Table 3, compound 13 exhibits greater in vivo activity compared to LY-3295668, and compounds of general formula (1) where L is of appropriate size, such as CD2, and / or W is [ka] This results in significantly improved efficacy in vivo, suggesting the potential for targeting cancer with Aurora kinase inhibitors.

[0154] Example 28: In vivo anti-tumor growth in H69 mouse xenografts H69 cells were cultured in 1640 medium containing 10% FBS at 37°C and 5% CO2. The cells were passaged and harvested by trypsinization. 1 × 10 7 H69 cells were implanted into the left axilla of nude mice. Mice were cultured until tumor volume reached approximately 290 mm 3 When the mice reached 100 mg / kg, they were randomly assigned to four groups of eight mice each and orally administered vehicle (0.5% MC) and 2.5 mg / kg, 5 mg / kg, and 10 mg / kg of Compound 1-1 twice daily. Tumor volume and body weight were measured every other day. Mice were sacrificed on the 21st day of treatment. Relative tumor volume (RTV) and tumor growth (T / C), as well as tumor growth inhibition (TGI), were calculated and statistically analyzed. The results are shown below.

[0155] Table 4: In vivo efficacy in a mouse H69 xenograft model

[0156] [Table 4]

[0157] *: P<0.05 vs. control, **: P<0.01 vs. control, ***: P<0.001 vs. control, ****: P<0.0001 vs. control; d1: first day of treatment; d21: last day of treatment; qd*21: once daily for 21 days; RTV: relative tumor volume; RTV=V t / V0; t / C(%)=t RTV / C RTV ×100; t RTV : relative tumor volume (RTV) of the treatment group; C RTV : Relative tumor volume (RTV) of the vehicle control group. TGI: tumor growth inhibition (%); T / C (%)>60%: no effect; T / C (%)≦60% and P<0.05: effective.

[0158] As shown in Table 4, compound 1-1 blocks H69 tumor growth in a dose-dependent manner with significantly improved efficacy compared to LY-3295668.

[0159] While specific embodiments of the present invention have been described above, it will be understood by those skilled in the art that these are merely examples and that various changes or modifications can be made to these embodiments without departing from the principles and spirit of the present invention. Accordingly, the scope of the present invention is defined by the appended claims.

Claims

1. A compound of formula (1), an optical isomer thereof, or a pharmaceutically acceptable salt thereof. 【Chemistry 1】 [In formula (1), R １ is aryl, heteroaryl 【Chemistry 2】 and The aryl and heteroaryl are optionally substituted with 1 to 3 groups selected from the group consisting of halogen, C1-C3 alkyl, C1-C3 alkoxyl, halogen-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxyl, or R 1 is 【Transformation 3】 and R ２ is H or methyl, R ３ is H or F, W is 【Chemistry 4】 and Here, R ａ is H, C1-C3 alkyl or C3-C6 cycloalkyl, Lは、CH ２ 、CO、CD ２ 、CH(Me),C(Me) ２ 、 【Transformation 5】 is]

2. In the formula (1), R １ teeth, 【Transformation 6】 [X is NH, O or S, and R ｃ and R ｄ are independently H, halogen, C1-C3 alkyl, C1-C3 alkoxyl, halogen-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxyl.

3. In the formula (1), R １ teeth, 【Transformation 7】 3. The compound of claim 2, wherein:

4. In the formula (1), W is 【Transformation 8】 2. The compound of claim 1, wherein:

5. The compound is 【Chemistry 9】 【change】 5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein:

6. 10. An Aurora kinase inhibitor comprising the compound according to any one of claims 1 to 5, an optical isomer thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

7. A combined pharmaceutical composition comprising a compound according to any one of claims 1 to 5 as an active ingredient and a pharmaceutically acceptable carrier or diluent.

8. 6. A method of using the compound of any one of claims 1 to 5, its optical isomer, or a pharmaceutically acceptable salt thereof, wherein the compound is used as an Aurora kinase inhibitor in the preparation of an antitumor medicament.

9. 6. A method for using the compound according to any one of claims 1 to 5, its optical isomer, or a pharmaceutically acceptable salt thereof, wherein the compound is used for preparing an antitumor drug.

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