TRPC6 inhibitor and preparation method therefor and use thereof

Abstract

The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof as a TRPC6 inhibitor, and a preparation method therefor. The compound or the pharmaceutically acceptable salt thereof can be used for the prevention or treatment of TRPC6-related diseases, including renal and urinary system diseases, cardiovascular diseases, neoplastic diseases, systemic dysfunction diseases, and the like. The present invention further provides a pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt thereof, and a use of the compound or the pharmaceutically acceptable salt thereof or the composition in the prevention or treatment of TRPC6-related diseases.

Description

TRPC6 Inhibitors and Their Preparation and Application Technical Field

[0001] This invention relates to TRPC6 inhibitor compounds, methods for their preparation, pharmaceutical compositions comprising such compounds, and the use of such compounds in the treatment of diseases. Specifically, this invention relates to amide derivatives containing nitrogen-containing heterocycles that can be used as TRPC6 inhibitors, methods for their preparation, pharmaceutical compositions comprising such derivatives, and the use of such derivatives in the prevention and treatment of TRPC6-related diseases. Background Technology

[0002] TRPC, or transient receptor potential cation channel, belongs to the TRP (Transient Receptor Potential) subfamily. It is a non-selective cation channel transmembrane protein on the cell membrane. TRPC channels consist of six transmembrane α-helices, with a hairpin-shaped cation channel structure formed by the 5th and 6th transmembrane domains. Each TRPC subunit contains a transmembrane region and adjacent intracellular N-terminus and C-terminus. These two ends are key sites for protein-protein interactions that coordinate channel transport, anchoring, localization, gating, and functional regulation. The C-terminus of a TRPC typically contains 3-6 ankyrin binding sites, while the ankyrin repeat sequence and coiled-coil region at the N-terminus are fundamental components of the tetrameric channel assembly. Ankyrin interaction with TRPC channels can inhibit intracellular calcium channel regulation by inositol phosphate receptor (IP3-R) and ryanodine receptor. 2+ release.

[0003] TRPC6, a member of the TRPC family, is encoded by the TRPC6 gene, containing 931 amino acids, and is expressed in organs such as the liver, kidneys, and lungs. The human TRPC6 gene is located on chromosome 11q21-22. Recent studies have shown that mutations or overexpression of the TRPC6 gene can cause abnormalities in intracellular calcium signaling pathways, leading to various pathophysiological changes, including podocyte damage, disruption of the glomerular filtration barrier, glomerulosclerosis, renal fibrosis, pulmonary vascular remodeling, pulmonary hypertension, lung inflammation and injury, myocardial hypertrophy, myocardial fibrosis, heart failure, hypertension, abnormal intracellular calcium signaling, tumor cell proliferation, tumor metastasis and invasion, tumor angiogenesis, and cardiovascular metabolic abnormalities. Therefore, research on TRPC6 as a therapeutic target has become a focus of attention.

[0004] In 2005, Winn et al. reported the discovery of a mutation in the TRPC6 gene in a large family of adult-onset focal segmental glomerulosclerosis (FSGS) with an autosomal dominant inheritance pattern. Researchers have found that TRPC6 expression is significantly increased in patients with non-hereditary proteinuric kidney diseases such as FSGS, minimal change disease (MCD), and membranous nephropathy (MN). TRPC6 channels are closely related to calcium ion concentration in the process of myocardial hypertrophy; its overexpression can increase intracellular calcium concentration in cardiomyocytes, becoming one of the important mechanisms inducing myocardial hypertrophy. Chu et al. cultured neonatal mouse cardiomyocytes under hypoxic conditions and found that hypoxia induced an increase in HIF-1α, thereby activating TRPC6 channels, leading to extracellular calcium influx, increasing intracellular calcium ion concentration, activating the nuclear factor of activated T cells (NFAT) signaling pathway, and subsequently causing cardiomyocyte remodeling and myocardial hypertrophy. Furthermore, recent studies have found that TRPC6 protein participates in changes in intracellular calcium concentration and is closely related to tumor occurrence and development as well as alterations in the tumor cell cycle. These research advances make TRPC6 protein a promising new target for cancer therapy. Summary of the Invention

[0005] In a first aspect, the present invention provides a small molecule inhibitor of TRPC6, which is a compound of general formula (I) or a pharmaceutically acceptable salt thereof.

[0006] in:

[0007] Ring A can be a 4-10 member nitrogen-containing heterocycle, spiroheterocycle, or bridged heterocycle, which is optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, amino, halogen, and -C(O)NH2, which are optionally substituted with amino groups.

[0008] The B ring can be a 6-10 membered monocyclic or bicyclic aromatic ring or a 6-10 membered monocyclic or bicyclic heteroaromatic ring containing a nitrogen atom, which may optionally be substituted by one or more substituents selected from the following: C1-C6 alkyl, C1-C6 alkoxy, halogen and cyano, and 5-membered heteroaryl containing 1-3 heteroatoms selected from N, S and O.

[0009] n can be 0 or 1;

[0010] R can be -C(O)-NR1R2 or -NR3-C(O)R4, wherein R1, R2, R3 and R4 can each be independently selected from C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 aryl C1-C3 alkylene and 5-8 heteroaryl C1-C3 alkylene, wherein the 5-8 heteroaryl can contain 1-3 heteroatoms selected from N, S and O, wherein the 6-10 aryl or 5-8 heteroaryl can be optionally substituted by one or more substituents selected from C1-C3 alkyl, halogen and cyano, or can form a 5-8 nitrogen-containing heterocyclic group fused with the benzene ring together with the attached nitrogen atom, wherein the benzene ring can be optionally substituted with halogen.

[0011] In one embodiment, in formula (I), R can be -C(O)-NR 1a R 2a , where R 1a and R 2a Each of the components can be independently selected from C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 aryl C1-C3 alkylene, and 5-8 heteroaryl C1-C3 alkylene, wherein the 5-8 heteroaryl contains 1-3 heteroatoms selected from N, S, and O, and the 6-10 aryl or 5-8 heteroaryl may optionally be substituted by one or more substituents selected from C1-C3 alkyl, halogen, and cyano. Alternatively, R... 1a and R 2a It can form a 5-8 membered nitrogen-containing heterocyclic group fused with a benzene ring together with the attached nitrogen atom, wherein the benzene ring can be optionally substituted with a halogen.

[0012] In this embodiment, R 1a It can be a C1-C6 alkyl or a C3-C6 cycloalkyl; R 2a It can be benzyl or 5-6-membered heteroarylmethylene, wherein the 5-6-membered heteroaryl contains 1-2 heteroatoms selected from N and S, wherein the benzyl or 5-6-membered heteroarylmethylene may optionally be substituted on the phenyl or 5-membered heteroaryl by one or more substituents selected from C1-C3 alkyl, halogen and cyano groups. Alternatively, R... 1a and R 2a It can form a 5- or 6-membered nitrogen-containing heterocyclic group fused with a benzene ring together with the attached nitrogen atom, wherein the benzene ring can be optionally substituted with F.

[0013] Preferably, R 1a It can be a C1-C3 alkyl or a C3-C6 cycloalkyl. More preferably, R 1a It can be methyl, isopropyl, or cyclopropyl. Most preferably, R... 1a It can be methyl.

[0014] Preferably, R 2aIt can be benzyl, pyridylmethylene, or thiazolylmethylene. More preferably, R 2a It can be benzyl.

[0015] In one embodiment, the compound of the present invention or a pharmaceutically acceptable salt thereof may be represented by the following formula (II):

[0016] The definitions of ring A and ring B can be the same as those in equation (I) above. In equation (II), R 1b It can be a C1-C6 alkyl or a C3-C6 cycloalkyl; R 2b It can be benzyl or 5-6-membered heteroarylmethylene, wherein the 5-6-membered heteroaryl contains 1-2 heteroatoms selected from N and S, wherein the benzyl or 5-6-membered heteroarylmethylene may optionally be substituted on the phenyl or 5-membered heteroaryl by one or more substituents selected from C1-C3 alkyl, halogen and cyano groups. Alternatively, R... 1b and R 2b It can form a 5- or 6-membered nitrogen-containing heterocyclic group fused with a benzene ring together with the attached nitrogen atom, wherein the benzene ring can be optionally substituted with F.

[0017] Preferably, R 1a It can be a C1-C3 alkyl or a C3-C6 cycloalkyl. More preferably, R 1a It can be methyl, isopropyl, or cyclopropyl. Most preferably, R... 1a It can be methyl.

[0018] Preferably, R 2a It can be benzyl, pyridylmethylene, or thiazolylmethylene. More preferably, R 2a It can be benzyl.

[0019] In any embodiment, ring A can be a nitrogen-containing heterocycle, spiroheterocycle, or bridged heterocycle selected from the following structures:

[0020] In any embodiment, ring B can be an aromatic ring or heteroaromatic ring selected from the following structures:

[0021] In a preferred embodiment, the TRPC6 inhibitor of the present invention may be a compound selected from the following or a pharmaceutically acceptable salt thereof:

[0022] The inventors have discovered that the compounds provided by this invention can effectively inhibit the activity of TRPC6, and therefore may be used for the prevention or treatment of TRPC6-related diseases. Specifically, given the crucial role of TRPC6 in related diseases, the compounds of this invention have the potential to become novel therapeutic agents for treating diseases related to abnormal TRPC6 expression. In particular, the compounds of this invention show potential clinical application prospects in the prevention and treatment of diseases such as kidney and urinary system diseases, cardiovascular system diseases, oncological diseases, and systemic functional disorders.

[0023] It is worth noting that, compared with other channels in the TRPC family such as TRPC3, TRPC5 and other calcium ion channels such as TRPV3, TRPM8, TRPA1, etc., the compounds of the present invention exhibit significant selectivity for TRPC6.

[0024] TRPC channels are widely expressed in various tissues of the human body and are closely related to numerous physiological functions. TRPC3 is highly expressed in the central nervous system and is involved in synaptic signal transmission and motor coordination. TRPC5 is mainly expressed in brain tissue, with some distribution in the liver and kidneys. It participates in various physiological processes and is associated with the generation of emotions such as fear, anxiety, and depression, as well as the occurrence of progressive kidney disease. TRPV3 plays an important role in the nervous system and sensory organs, especially in touch and temperature perception. TRPM8 is expressed even more widely, not only in peripheral dorsal root ganglia and central trigeminal neurons, but also in vascular smooth muscle, lungs, skeletal muscle, mesentery, gastric fundus, liver, bladder, prostate, breast, thymus, and skin. TRPA1 is mainly expressed in the terminals of small- to medium-diameter primary sensory neurons and relies on nociceptors to perceive harmful stimuli in the environment. Unlike the above ion channels, TRPC6 is expressed at high levels in organs such as the heart, kidneys, and lungs, and plays important physiological functions in these organs.

[0025] Therefore, the significant selectivity of the compounds of the present invention for TRPC6 enables the application of the compounds of the present invention to reduce the impact on other non-targeted channels, reduce interference with normal physiological functions of the body, thereby reducing the risk of potential side effects and improving the safety of treatment.

[0026] The compounds described herein can be prepared and / or used as pharmaceutically acceptable salts. These pharmaceutically acceptable salts can be inorganic acid salts or organic acid salts. The inorganic acid salts can be salts formed with hydrohalic acids, nitric acid, carbonic acid, sulfuric acid, phosphoric acid, etc. The organic acid salts can be salts formed with malic acid, citric acid, fumaric acid, oxalic acid, lactic acid, camphorsulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, benzoic acid, etc. The hydrohalic acids can be hydrofluoric acid, hydrobromic acid, hydroiodic acid, or hydrochloric acid. These pharmaceutically acceptable salts can be prepared by methods well known to those skilled in the art.

[0027] When the compounds of the present invention have isomers, such as optical isomers, stereoisomers, positional isomers, and rotational isomers, mixtures of any of the isomers are included within the scope of the compounds of the present invention. For example, when the compounds of the present invention have optical isomers, optical isomers separated from racemic mixtures are also included within the scope of the compounds of the present invention. Each of these isomers can be obtained as a separate compound by known synthetic and separation methods.

[0028] The compounds or salts thereof of the present invention may be amorphous or crystalline. In the case of crystals, both single crystals and polymorphs are included within the scope of the compounds or salts thereof of the present invention. These crystals may be produced by crystallization using methods known per se in the art.

[0029] The compounds or salts thereof of the present invention may be in the form of solvates or non-solvents. For example, the compounds or salts thereof of the present invention may be in the form of hydrates. Any of these forms is included within the scope of the compounds or salts thereof of the present invention.

[0030] The compounds of the present invention or their salts may be made using isotopes (e.g., 2 H(D), 3 H, 13 C 14 C 35 S, 125 I) Forms of labeling. In the case of isotopic labeling, the isotopic atom can be an atom of a naturally occurring, but not most common, isotopic element. For example, the deuterium abundance level of the compounds of the present invention or their salts is greater than the naturally occurring abundance of deuterium. Any of these forms are included within the scope of the compounds of the present invention or their salts. Isotopic-labeled compounds, such as deuterated compounds, can be prepared by methods known per se in the art.

[0031] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof according to a first aspect of the invention, and one or more pharmaceutically acceptable excipients.

[0032] The pharmaceutically acceptable excipients mentioned above can be diluents, fillers, binders, humectants, disintegrants, absorption promoters, surfactants, adsorbents, lubricants, flavorings, sweeteners, etc., which are commonly used in the pharmaceutical field.

[0033] The pharmaceutical compositions of the present invention can be formulated according to techniques known in the art. The pharmaceutical compositions can be in any form suitable for oral, parenteral, topical, intranasal, intrabronchial, sublingual, ocular, ocular, rectal, vaginal, or transdermal administration. Preferably, the pharmaceutical compositions of the present invention can be in the form of oral administration.

[0034] In a fourth aspect, the present invention provides the use of a compound or a pharmaceutically acceptable salt thereof according to the first aspect of the invention for the prevention or treatment of TRPC6-related diseases.

[0035] The present invention provides, in a fifth aspect, a method for preventing or treating TRPC6-related diseases, comprising administering a compound or a pharmaceutically acceptable salt thereof according to a first aspect of the invention to a subject in need of such treatment.

[0036] The invention provides, in a sixth aspect, the use of a compound or a pharmaceutically acceptable salt thereof according to the first aspect of the invention, or a composition according to the second aspect of the invention, in the manufacture of a medicament for the prevention or treatment of TRPC6-related diseases.

[0037] The dosage forms of the aforementioned drugs can include tablets, capsules, patches, ointments, emulsions, suspensions, gels, powders, granules, oral liquids, and injections. Preferably, the dosage form of the drug can be a dosage form suitable for oral administration. These dosage forms can all be manufactured according to conventional methods in the pharmaceutical field.

[0038] In this invention, TRPC6-related diseases may include, but are not limited to, kidney and urinary system diseases, cardiovascular system diseases, neoplastic diseases, and systemic functional disorders.

[0039] TRPC6-related kidney and urinary system diseases can include, but are not limited to, glomerular dysfunction diseases (e.g., focal segmental glomerulosclerosis, membranous nephropathy, and IgA nephropathy), metabolic nephropathy (e.g., diabetic nephropathy and hypertensive nephropathy), acute / chronic kidney injury, tubulointerstitial lesions, and secondary renal fibrosis. TRPC6-related cardiovascular diseases can include, but are not limited to, circulatory dysfunction diseases (e.g., primary and secondary hypertension and pulmonary hypertension), myocardial remodeling-related diseases (e.g., heart failure, myocardial hypertrophy, and myocarditis), vascular dysfunction diseases (e.g., atherosclerosis and vascular calcification), arrhythmia diseases (e.g., various arrhythmias), and thromboembolic diseases. TRPC6-related neoplastic diseases can include, but are not limited to, solid tumors associated with abnormal TRPC6 expression (e.g., gastric adenocarcinoma, lung cancer, breast cancer, and prostate cancer) and hematologic malignancies. TRPC6-related systemic dysfunctions can include respiratory diseases (such as asthma, chronic obstructive pulmonary disease, and idiopathic pulmonary fibrosis) and metabolic disorders (such as insulin resistance syndrome and non-alcoholic steatohepatitis). Attached Figure Description

[0040] Figures 1 and 2 show undifferentiated and differentiated MPC5 cells from test example 3, respectively.

[0041] Figure 3 shows the relative cell viability after treatment with different concentrations of the agonist M085 for 48 hours;

[0042] Figure 4 shows the fluorescence intensity of Fluo-4 after treatment with agonist M085 for different times;

[0043] Figure 5 shows the relative cell viability after 48 hours of treatment with M085 alone and with M085 and different concentrations of compound 25 simultaneously;

[0044] Figures 6 and 7 show the drug-time curves of compound 16 in SD rat plasma administered via intravenous injection and oral gavage, respectively. Detailed Implementation

[0045] The present invention will be further described in conjunction with the embodiments. The following embodiments are only illustrative of the present invention and are not intended to limit the present invention in any way.

[0046] Compound Synthesis

[0047] The compounds of the present invention are synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art. Generally, the synthesis of the compounds disclosed herein can utilize reactions known in the art, and these reactions can be modified by reagents and reaction conditions deemed appropriate by those skilled in the art to introduce various parts of the molecules provided herein.

[0048] If necessary, the reaction products can be separated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, and chromatography. These products can be characterized using conventional methods, including physical constants and spectral data.

[0049] In the following preparation examples 1-44, all reagents used are conventional unless otherwise specified; all synthetic methods are conventional unless otherwise specified. Column chromatography was performed using silica gel (FCP 200-300 mesh) obtained from Sinopharm Chemical Reagent Co., Ltd. NMR spectra were measured using an ECZ-400S / L1 nuclear magnetic resonance spectrometer obtained from NEC Corporation, with deuterated chloroform (CDCl3) or deuterated dimethyl sulfoxide (DMSO-d6) as the solvent, and tetramethylsilane (TMS) as an internal standard; otherwise, the NMR solvent was used as an internal standard. All δ values ​​are expressed in ppm. MS measurements were performed using an IQ mass spectrometer obtained from Agilent Technologies.

[0050] The abbreviations used in the following synthesis steps are explained below:

[0051] Preparation Example 1. Synthesis of 2-(4-aminopiperidin-1-yl)-N-benzyl-N-methylbenzamide (Compound 1)

[0052] Step 1. Synthesis of intermediate 1a of methyl 2-(4-((tert-Butoxycarbonyl)amino)piperidin-1-yl)benzoate

[0053] In a 100 mL three-necked flask, methyl 2-fluorobenzoate (2.0 g, 13.0 mmol) dissolved in DMF (20 mL) was added, followed by 4-tert-butoxycarbonylaminopiperidine (2.8 g, 14.3 mmol) and K₂CO₃ (5.4 g, 39.0 mmol). The mixture was heated to 100 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain an oily product 1a (2.2 g, 50% yield). ESI-MS: m / z = 335 [M+1] + .

[0054] Step 2. Synthesis of intermediate 1b of 2-(4-((tert-Butoxycarbonyl)amino)piperidin-1-yl)benzoic acid

[0055] In a 100 mL three-necked flask, 1a (2.0 g, 6.0 mmol) dissolved in EtOH (20 mL) was added, followed by 2N NaOH (10 mL, 20.0 mmol). The mixture was heated to 60 °C and magnetically stirred for 3 h. After the reaction was complete, the mixture was cooled to room temperature, and water and ethyl acetate were added. The pH was adjusted to 2 with ice-cold 1N HCl. The mixture was separated into liquid and liquid phases. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined. The mixture was washed with water and saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to give solid 1b (1.4 g, 73% yield). ESI-MS: m / z = 321 [M+1] + .

[0056] Step 3. Synthesis of intermediate 1c of (1-(2-(benzyl(methyl)carbamoyl)phenyl)piperidin-4-yl)carbamate tert-butyl ester

[0057] In a 100 mL three-necked flask, 1c (1.3 g, 4.1 mmol) dissolved in DCM (15 mL) was added, followed by N-methylbenzylamine (0.6 g, 5.3 mmol), EDCI (1.0 g, 5.3 mmol), and HoBt (0.7 g, 5.3 mmol). The mixture was magnetically stirred at room temperature for 16 h. After the reaction was complete, water was added, and the mixture was extracted with dichloromethane. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give an oily 1c (1.4 g, 81% yield). ESI-MS: m / z = 424 [M+1] + .

[0058] Step 4. Synthesis of 2-(4-aminopiperidin-1-yl)-N-benzyl-N-methylbenzamide (Compound 1)

[0059] Add 1c (1.3g, 3.1mmol) dissolved in DCM (15ml) to a 100ml three-necked flask, add TFA (2ml), and stir magnetically at room temperature for 3h. After the reaction is complete, add water, adjust the pH to 11 with 2N NaOH, extract with dichloromethane, combine the organic phases, wash with water, dry with anhydrous sodium sulfate, evaporate to dryness using a rotary evaporator, and purify the residue by silica gel column chromatography (PE:EA = 1:1, v / v) to give compound 1 (0.8g, yield 80%) as a white solid.

[0060] 1H NMR (400MHz, CDCl3): δ8.10(s,2H),7.76-7.60(m,2H),7.59-7.30(m,4H),7.22-7.12(m,1H),4.79-4.49(m,2H) ),3.97-3.78(m,3H),3.66(d,J=1.0Hz,2H),3.21-2.87(m,3H),2.40(t,J=15.8Hz,4H).ESI-MS:m / z=324[M+1] + .

[0061] Preparation Example 2. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methylbenzamide (Compound 2)

[0062] Step 1. Synthesis of methyl 2-(4-((tert-butoxycarbonyl)amino)azapyro-1-yl)benzoate (intermediate 2a)

[0063] The synthesis of 2a was performed using the same method as 1a, except that 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 2a (2.3 g, 51% yield). ESI-MS: m / z = 349 [M+1] + .

[0064] Step 2. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)benzoic acid (intermediate 2b)

[0065] The synthesis method for 2b is the same as that for 1b, except that 1a is replaced with 2a, yielding solid 2b (2.0 g, 91% yield). ESI-MS: m / z = 335 [M+1] + .

[0066] Step 3. Synthesis of (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)phenyl)azapor-4-yl)tert-butyl carbamate (intermediate 2c)

[0067] The synthesis method for 2c is the same as that for 1c, except that 1b is replaced with 2b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 2c (1.6 g, yield 58%). ESI-MS: m / z = 456 [M+1] + .

[0068] Step 4. Synthesis of 2-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylbenzamide (compound 2)

[0069] The synthesis method of compound 2 is the same as that of compound 1, except that 1c is replaced with 2c, to obtain compound 2 (0.9g, yield 72%), which is a white solid.

[0070] 1 H NMR (400MHz, CDCl3): δ7.82-7.65(m,3H),7.54(dd,J=14.9,7.8Hz,2H),7.46-7.30(m,1H),7.25-6.96(m,2H),4.82(dd,J=37.9,14.1Hz,2H),3 .78(t,J=10.6Hz,5H),3.08(d,J=36.5Hz,3H),2.56(s,2H),2.41(d,J=13.9Hz,2H),2.25(s,1H),2.10(d,J=8.6Hz,1H).ESI-MS: m / z=356[M+1] + .

[0071] Preparation Example 3. Synthesis of 2-(4-aminoachrogen-1-yl)-N-(4-fluorobenzyl)-N-methylbenzamide (Compound 3)

[0072] Step 1. Synthesis of (1-(2-(4-fluorobenzyl)(methyl)carbamoyl)phenyl)azapor-4-yl)tert-butyl carbamate (intermediate 3c)

[0073] The synthesis method for 3c is the same as that for 1c, except that 1b is replaced with 2b and N-methylbenzylamine is replaced with N-methyl-4-fluorobenzylamine, yielding oily 3c (1.8 g, yield 65%). ESI-MS: m / z = 456 [M+1] + .

[0074] Step 2. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(4-fluorobenzyl)-N-methylbenzamide (compound 3)

[0075] The synthesis method of compound 3 is the same as that of compound 1, except that 1c is replaced with 3c, to obtain white solid 3 (0.7g, yield 48%).

[0076] 1H NMR (400MHz, CDCl3): δ7.91 (s, 2H), 7.77-7.60 (m, 2H), 7.57-7.38 (m, 2H), 7.31 (dd, J = 8.3, 5.2Hz, 1H), 7.22-6.98 (m, 1H), 4.71 (dd, J = 72. 9,13.6Hz,2H),3.99-3.62(m,5H),3.08(d,J=14.5Hz,3H),2.51(s,2H),2.28(d,J=59.3Hz,3H),2.08-1.94(m,1H).ESI-MS:m / z=356[M+1] + .

[0077] Preparation Example 4. Synthesis of 2-(4-aminoachaplon-1-yl)-3-fluoro-N-(2-fluorobenzyl)-N-methylbenzamide (Compound 4)

[0078] Step 1. Synthesis of methyl 2-(4-((tert-butoxycarbonyl)amino)azapyro-1-yl)-3-fluorobenzoate (intermediate 4a)

[0079] The synthesis of 4a was performed using the same method as 1a, except that methyl 2-fluorobenzoate was replaced with methyl 2,3-difluorobenzoate, and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 4a (2.6 g, 62% yield). ESI-MS: m / z = 367 [M+1] + .

[0080] Step 2. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-3-fluorobenzoic acid (intermediate 4b)

[0081] The synthesis method for 4b is the same as that for 1b, except that 1a is replaced with 4a, yielding solid 4b (2.0 g, 80% yield). ESI-MS: m / z = 353 [M+1] + .

[0082] Step 3. Synthesis of (1-(2-fluoro-6-(2-fluorobenzyl)(methyl)carbamoyl)phenyl)azapyrrolidone-4-yl)tert-butyl carbamate (intermediate 4c)

[0083] The synthesis method for 4c is the same as that for 1c, except that 1b is replaced with 4b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 4c (1.5 g, yield 56%). ESI-MS: m / z = 474 [M+1] + .

[0084] Step 4. Synthesis of 2-(4-aminoachaplon-1-yl)-3-fluoro-N-(2-fluorobenzyl)-N-methylbenzamide (compound 4)

[0085] The synthesis method of compound 4 is the same as that of compound 1, except that 1c is replaced with 4c, to obtain compound 4 (0.8g, yield 67%), which is a white solid.

[0086] 1 H NMR (400MHz, CDCl3): δ7.52(s,1H),7.44–7.28(m,3H),7.25-7.00(m,3H),4.85(dd,J=138.8,14.5Hz,1H),4.64-4.51(m,1H ),3.59(dd,J=111.1,44.5Hz,5H),3.04(d,J=47.1Hz,3H),2.29(d,J=15.8Hz,2H),2.10-1.77(m,4H).ESI-MS:m / z=374[M+1] + .

[0087] Preparation Example 5. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-5-methoxy-N-methylbenzamide (Compound 4)

[0088] Step 1. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-5-methoxybenzoic acid (intermediate 5a)

[0089] In a 100 mL three-necked flask, 2-bromo-5-methoxybenzoic acid (1.0 g, 4.3 mmol) dissolved in water (20 mL) and isopropanol (5 mL) were added. Then, 4-tert-butoxycarbonyl-1H-azapyridine (1.3 g, 6.5 mmol), K₂CO₃ (1.2 g, 8.6 mmol), KOH (0.48 g, 8.6 mmol), and Cu (0.03 g, 2%) were added. The mixture was heated to 105 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and the pH was adjusted to 1 with ice-cold 1N HCl. The mixture was extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to give a white solid, 5a (0.9 g, 57% yield). ESI-MS: m / z = 365 [M+1] + .

[0090] Step 2. Synthesis of tert-butyl carbamate (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-4-methoxyphenyl)azapyrrolidone-4-yl)carbamate (intermediate 5b)

[0091] The synthesis method for 5b is the same as that for 1c, except that 1b is replaced with 5a, yielding oily 5b (0.7 g, yield 56%). ESI-MS: m / z = 486 [M+1] + .

[0092] Step 3. Synthesis of 2-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-5-methoxy-N-methylbenzamide (compound 5)

[0093] The synthesis method of compound 5 is the same as that of compound 1, except that 1c is replaced with 5b, to obtain a grayish-white solid compound 5 (0.1 g, yield 23%).

[0094] 1 H NMR (400MHz, CDCl3): δ7.82–7.65(m,3H),7.54(dd,J=14.9,7.8Hz,2H),7.46–7.30(m,1H),7.25–6.96(m,1H),4.82(dd,J=37.9,14.1Hz,2H),3.78(t ,J=10.6Hz,3H),3.55(s,3H),3.08(d,J=36.5Hz,3H),2.56(s,2H),2.41(d ,J=13.9Hz,2H),2.25(s,1H),2.10(d,J=8.6Hz,1H).ESI-MS:m / z=386[M+1] + .

[0095] Preparation Example 6. Synthesis of 2-(4-aminoachrogen-1-yl)-3-cyano-N-(2-fluorobenzyl)-N-methylbenzamide (Compound 6)

[0096] Step 1. Synthesis of methyl 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-3-cyanobenzoate (intermediate 6a)

[0097] The synthesis of 6a was performed using the same method as 1a, except that methyl 2-fluorobenzoate was replaced with methyl 3-cyano-2-fluorobenzoate, and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 6a (3.3 g, 80% yield). ESI-MS: m / z = 374 [M+1] + .

[0098] Step 2. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-3-cyanobenzoic acid (intermediate 6b)

[0099] The synthesis method for 6b is the same as that for 1b, except that 1a is replaced with 6a, yielding a white solid 6b (2.2 g, 70% yield). ESI-MS: m / z = 360 [M+1] + .

[0100] Step 3. Synthesis of (1-(2-cyano-6-(2-fluorobenzyl)(methyl)carbamoyl)phenyl)azapyrrolidone-4-yl)tert-butyl carbamate (intermediate 6c)

[0101] The synthesis method for 6c is the same as that for 1c, except that 1b is replaced with 6b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 6c (2.3 g, yield 78%). ESI-MS: m / z = 481 [M+1] + .

[0102] Step 4. Synthesis of 2-(4-aminoacrahepane-1-yl)-3-cyano-N-(2-fluorobenzyl)-N-methylbenzamide (compound 6)

[0103] The synthesis method of compound 6 is the same as that of compound 1, except that 1c is replaced with 6c, to obtain a grayish-white solid compound 6 (0.9 g, yield 51%).

[0104] 1 H NMR (400MHz, CDCl3): δ7.68-7.51(m,2H),7.49-7.32(m,2H),7.25-7.00(m,3H),4.84(dt,J=80.5,15.0Hz,1 H),3.70-3.24(m,4H),3.21-2.97(m,2H),2.87(d,J=27.9Hz,2H),2.04-1.65(m,7H).ESI-MS:m / z=381[M+1] + .

[0105] Preparation Example 7. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,4-triazol-1-yl)benzamide (compound 7)

[0106] Step 1. Synthesis of methyl 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-4-iodobenzoate (intermediate 7a)

[0107] The synthesis method for 7a was the same as that for 1a, except that methyl 2-fluorobenzoate was replaced with methyl 2-fluoro-4-iodobenzoate, and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 7a (2.8 g, yield 82%). ESI-MS: m / z = 475 [M+1] + .

[0108] Step 2. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-4-iodobenzoic acid (intermediate 7b)

[0109] The synthesis method for 7b is the same as that for 1b, except that 1a is replaced with 7a, yielding solid 7b (2.3 g, yield 87%). ESI-MS: m / z = 460 [M+1] + .

[0110] Step 3. Synthesis of tert-butyl carbamate (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-5-iodophenyl)azapyrrolidone-4-yl)carbamate (intermediate 7c)

[0111] The synthesis method for 7c is the same as that for 1c, except that 1b is replaced with 7b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 7c (2.0 g, yield 66%). ESI-MS: m / z = 581 [M+1] + .

[0112] Step 4. Synthesis of tert-butyl carbamate (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-5-(1H-1,2,4-triazol-1-yl)phenyl)azapyrrol-4-yl)carbamate (intermediate 7d)

[0113] In a 100 mL three-necked flask, 7c (1.0 g, 1.7 mmol) dissolved in DMF (10 mL) was added, followed by 1,2,4-triazole (0.4 g, 5.1 mmol), CuI (0.1 g, 10%), N,N-dimethyl-1,2-cyclohexanediamine (0.2 g, 20%), and Cs₂CO₃ (1.6 g, 5.1 mmol). The mixture was heated to 115 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel chromatography (EA) to give 7d (0.3 g, 34% yield) as a solid. ESI-MS: m / z = 523 [M+1] + .

[0114] Step 5.2 Synthesis of (4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,4-triazol-1-yl)benzamide (compound 7)

[0115] The synthesis method of compound 7 is the same as that of compound 1, except that 1c is replaced with 7d, to obtain a pale yellow solid compound 7 (0.2 g, yield 79%).

[0116] 1 H NMR (400MHz, CDCl3): δ8.12(s,1H),7.61-7.50(m,1H),7.43-7.25(m,3H),7.24-6.96(m,4H),4.84(ddd,J=49.9,33.6,14.4Hz, 1H),4.63-4.27(m,1H),3.65-3.18(m,3H),3.16-2.99(m,2H),2.87(d,J=19.0Hz,2H),2.14-1.56(m,7H).ESI-MS:m / z=423[M+1] + .

[0117] Preparation Example 8. Synthesis of 3-(4-aminoachrogen-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 8)

[0118] Step 1. Synthesis of 3-iodo-2-naphthoic acid (intermediate 8a)

[0119] 3-Amino-2-naphthoic acid (0.5 g, 2.7 mmol) was dissolved in concentrated HCl (20 ml) and H₂O (20 ml) in a 100 ml three-necked flask. The mixture was cooled to 0 °C, and NaNO₂ (0.7 g, 10.1 mmol) was added in portions. The mixture was stirred at 0 °C for 1 h. KI (5.3 g, 31.9 mmol) was added in portions, and the mixture was slowly heated to room temperature, then heated to 70 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain an oily 8a (0.9 g), which was used directly in the next reaction without purification. ESI-MS: m / z = 299 [M+1] + .

[0120] Step 2. Synthesis of methyl 3-iodo-2-naphthoate (intermediate 8b)

[0121] In a 100 mL three-necked flask, 8a (0.9 g, 3.0 mmol) was dissolved in DCM (10 mL), CDI (0.7 g, 4.5 mmol), and MeOH (2 mL). The mixture was magnetically stirred at room temperature for 16 h. After the reaction was complete, water was added for dilution, and the mixture was extracted with dichloromethane. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 10:1, v / v) to give oily 8b (0.8 g, 95% yield in two steps). ESI-MS: m / z = 312 [M+1] + .

[0122] Step 3. Synthesis of methyl 3-(4-((tert-butoxycarbonyl)amino)azacycloheptan-1-yl)-2-naphthoic acid (intermediate 8c)

[0123] In a 100 mL three-necked flask, 8b (0.8 g, 2.6 mmol) dissolved in toluene (10 mL) was added, followed by 4-tert-butoxycarbonyl-1H-azapyrrolidone (0.7 g, 3.3 mmol), Pd₂(DBA)₃ (0.1 g, 10%) / Ruphos (0.1 g, 10%), and Cs₂CO₃ (2.5 g, 7.7 mmol). The air was replaced three times with N₂, and the mixture was heated to 110 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 7:1, v / v) to give oily 8c (150 mg, 14% yield). ESI-MS: m / z = 399 [M+1] + .

[0124] Step 4. Synthesis of 3-(4-((tert-butoxycarbonyl)amino)azacycloheptane-1-yl)-2-naphthoic acid (intermediate 8d)

[0125] The synthesis method for 8d is the same as that for 1b, except that 1a is replaced with 8c, yielding solid 8d (0.1 g, 70% yield). ESI-MS: m / z = 385 [M+1] + .

[0126] Step 5. Synthesis of tert-butyl carbamate (intermediate 8e) of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)azacycloheptane-4-yl)carbamate

[0127] The synthesis method for 8e is the same as that for 1c, except that 1b is replaced with 8d and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 8e (0.03 g, yield 20%). ESI-MS: m / z = 481 [M+1] + .

[0128] Step 6. Synthesis of 3-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 8)

[0129] The synthesis method of compound 8 is the same as that of compound 1, except that 1c is replaced with 8e, to obtain oily compound 8 (0.016 g, yield 74%).

[0130] 1 H NMR (400MHz, CDCl3): δ7.74 (ddd, J=50.3, 35.4, 23.9Hz, 4H), 7.45 (t, J=7.5 Hz,1H),7.39-7.27(m,2H),7.22(t,J=7.3Hz,1H),7.15-6.91(m,2H),4.88(d dd,J=50.6,35.6,14.5Hz,1H),4.42(ddd,J=20.7,15.9,6.4Hz,1H),3.64-3 .14(m,6H),2.82(d,J=6.4Hz,2H),2.17-1.66(m,6H).ESI-MS:m / z=406[M+1] + .

[0131] Preparation Example 9. Synthesis of 2-(4-aminoachrogen-1-yl)-N-benzyl-3-fluoro-N-(2-fluorobenzyl)benzamide (Compound 9)

[0132] Step 1. Synthesis of N-benzyl-2-fluorobenzylamine (intermediate 9a)

[0133] 2-Fluorobenzamine (1.0 g, 5.8 mmol) dissolved in DMF (10 ml) was added to a 100 ml three-necked flask. Benzyl bromide (0.8 g, 6.4 mmol) and K₂CO₃ (2.4 g, 17.5 mmol) were then added. The mixture was magnetically stirred at room temperature for 2 h. After the reaction was complete, water was added to dilute the solution, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by column chromatography (PE:EA = 4:1, v / v) to give an oily 9a (0.7 g, 56% yield). ESI-MS: m / z = 216 [M+1] + .

[0134] Step 2. Synthesis of tert-butyl (2-(benzyl(2-fluorobenzyl)carbamoyl)-6-fluorophenyl)azapyrrolidone-4-yl)carbamate (intermediate 9b)

[0135] The synthesis method of 9b is the same as that of 1c, except that 1b is replaced with 4b and N-methylbenzylamine is replaced with 9a, yielding oily 9b (1.0 g, yield 59%). ESI-MS: m / z = 550 [M+1] + .

[0136] Step 3. Synthesis of 2-(4-aminoachaplon-1-yl)-N-benzyl-3-fluoro-N-(2-fluorobenzyl)benzamide (compound 9)

[0137] The synthesis method of compound 9 is the same as that of compound 1, except that 1c is replaced with 9b, to obtain oily compound 9 (0.5g, yield 62%).

[0138] 1 H NMR (400MHz, CDCl3): δ7.44–7.23(m,5H),7.24–6.92(m,6H),5.15–4.68(m,1H),4.65–4.16( m,4H),3.11(d,J=91.6Hz,4H),2.17–1.49(m,3H),1.38–1.19(m,3H).ESI-MS:m / z=450[M+1] + .

[0139] Preparation Example 10. Synthesis of 2-(4-aminoachaplon-1-yl)-3-fluoro-N-(2-fluorobenzyl)-N-isopropylbenzamide (Compound 10)

[0140] Step 1. Synthesis of N-isopropyl-2-fluorobenzylamine (intermediate 10a)

[0141] The synthesis method for 10a is the same as that for 9a, except that benzyl bromide is replaced with 2-bromopropane, yielding oily 10a (1.3 g, 50% yield). ESI-MS: m / z = 168 [M+1] + .

[0142] Step 2. Synthesis of tert-butyl (2-(isopropyl(2-fluorobenzyl)carbamoyl)-6-fluorophenyl)azapyrrolidone-4-yl)carbamate (intermediate 10b)

[0143] The synthesis method for 10b is the same as that for 1c, except that 1b is replaced with 4b and N-methylbenzylamine is replaced with 10a, yielding oily 10b (2.5 g, yield 62%). ESI-MS: m / z = 502 [M+1] + .

[0144] Step 3. Synthesis of 2-(4-aminoachaplon-1-yl)-N-isopropyl-3-fluoro-N-(2-fluorobenzyl)benzamide (Compound 10)

[0145] The synthesis method of compound 10 is the same as that of compound 1, except that 1c is replaced with 10b, to obtain oily compound 10 (1.4g, yield 70%).

[0146] 1 H NMR (400MHz, CDCl3): δ8.12 (d, J = 7.8Hz, 1H), 7.56-7.43 (m, 1H), 7.28-7.0 3(m,5H),4.74(qt,J=36.6,18.5Hz,2H),3.34-3.15(m,4H),3.02-2.80(m,2 H),2.07-1.91(m,3H),1.81(s,1H),1.62(d,J=7.7Hz,2H),1.47(dd,J=14. 0,8.4Hz,1H),1.36-1.21(m,1H),1.18-1.08(m,4H).ESI-MS:m / z=402[M+1] + .

[0147] Preparation Example 11. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-5-(1H-1,2,3-triazol-1-yl)benzamide (compound 11)

[0148] Step 1. Synthesis of methyl 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-5-iodobenzoate (intermediate 11a)

[0149] The synthesis of 11a was performed using the same method as that of 1a, except that methyl 2-fluorobenzoate was replaced with methyl 2-fluoro-5-iodobenzoate, and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 11a (2.0 g, yield 58%). ESI-MS: m / z = 475 [M+1] + .

[0150] Step 2. Synthesis of 2-(4-((tert-Butoxycarbonyl)amino)azapyro-1-yl)-5-iodobenzoic acid (intermediate 11b)

[0151] The synthesis method for 11b is the same as that for 1b, except that 1a is replaced with 11a, yielding solid 11b (1.8 g, 95% yield). ESI-MS: m / z = 460 [M+1] + .

[0152] Step 3. Synthesis of tert-butyl (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-4-iodophenyl)azapyrrolidone-4-yl)carbamate (intermediate 11c)

[0153] The synthesis method for 11c is the same as that for 1c, except that 1b is replaced with 11b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 11c (2.0 g, yield 86%). ESI-MS: m / z = 581 [M+1] + .

[0154] Step 4. Synthesis of tert-butyl carbamate (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-4-(1H-1,2,4-triazol-1-yl)phenyl)azapyrrol-4-yl)carbamate (intermediate 11d)

[0155] The synthesis method for 11d is the same as that for 7d, except that 7c is replaced with 11c and 1,2,4-triazole is replaced with 1,2,3-triazole, yielding oily 11d (0.6 g, yield 34%). ESI-MS: m / z = 523 [M+1] + .

[0156] Step 5. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-5-(1H-1,2,4-triazol-1-yl)benzamide (compound 11)

[0157] The synthesis method of compound 11 is the same as that of compound 1, except that 1c is replaced with 11d, to obtain oily compound 11 (0.4g, yield 79%).

[0158] 1 H NMR (400MHz, CDCl3): δ7.95(d,J=8.2Hz,1H),7.85(d,J=2.6Hz,1H),7.76(d,J=7.9Hz,2H),7.52(s,1H),7.25-6.88(m,4H),4.7 1-4.34(m,1H),3.70-3.15(m,4H),3.07(t,J=11.7Hz,1H),2.87(s,2H),1.90(dd,J=106.0,62.5Hz,8H).ESI-MS:m / z=423[M+1] + .

[0159] Preparation Example 12. Synthesis of 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,3-triazol-1-yl)benzamide (Compound 12)

[0160] Step 1. Synthesis of tert-butyl carbamate (1-(2-(2-fluorobenzyl)(methyl)carbamoyl)-4-(1H-1,2,4-triazol-1-yl)phenyl)azapyrrol-4-yl)carbamate (intermediate 12b)

[0161] The synthesis method for 12b is the same as that for 7d, except that 1,2,4-triazole is replaced with 1,2,3-triazole, yielding oily 12b (0.3 g, yield 33%). ESI-MS: m / z = 523 [M+1] + .

[0162] Step 2.2 Synthesis of (4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,3-triazol-1-yl)benzamide (compound 12)

[0163] The synthesis method of compound 12 is the same as that of compound 1, except that 1c is replaced with 12b, to obtain oily compound 12 (0.14 g, yield 59%).

[0164] 1 H NMR (400MHz, CDCl3): δ7.95(d,J=8.2Hz,1H),7.85(d,J=2.6Hz,1H),7.76(d,J=7.9Hz,2H),7.25-6.88(m,4H),7.15(s,1H),4.7 1-4.34(m,1H),3.70-3.15(m,4H),3.07(t,J=11.7Hz,1H),2.87(s,2H),1.90(dd,J=106.0,62.5Hz,8H).ESI-MS:m / z=423[M+1] + .

[0165] Preparation Example 13. Synthesis of 2-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-3-carboxamide (Compound 13)

[0166] Step 1. Synthesis of 2-chloroquinoline-3-carboxylic acid (intermediate 13a)

[0167] In a 250 mL three-necked flask, 2-chloro-3-quinoline carbaldehyde (1.0 g, 5.2 mmol) dissolved in 1,4-dioxane (50 mL) and H₂O (50 mL) was added. The mixture was cooled to 0 °C, and H₃NO₃S (3.0 g, 31.3 mmol) was added. Then, NaClO₂ (0.8 g, 6.4 mmol) and KH₂PO₄ (8.5 g, 62.6 mmol) were added in portions. The mixture was magnetically stirred at room temperature for 16 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to give a white solid, 13a (1.1 g, 102% yield). ESI-MS: m / z = 208 [M+1] + .

[0168] Step 2. Synthesis of 2-chloro-N-(2-fluorobenzyl)-N-methylquinoline-3-carboxamide (intermediate 13b)

[0169] The synthesis method for 13b is the same as that for 1c, except that 1b is replaced with 13a and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 13b (1.0 g, yield 57%). ESI-MS: m / z = 329 [M+1] + .

[0170] Step 3. Synthesis of tert-butyl carbamate (intermediate 13c) of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)quinoline-2-yl)azacycloheptyl-4-yl)carbamate

[0171] In a 100 mL three-necked flask, 13b (1.0 g, 3.0 mmol) dissolved in DMSO (10 mL) was added, followed by 4-tert-butoxycarbonyl-1H-azapyrrolidone (1.0 g, 4.5 mmol), DIPEA (1.2 g, 9.0 mmol), KF (0.3 g, 6.0 mmol), and CuI (0.1 g, 10%). The mixture was heated to 135 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel chromatography (PE:EA = 1:1, v / v) to give oily 13c (1.4 g, 92% yield). ESI-MS: m / z = 507 [M+1] + .

[0172] Step 4. Synthesis of 2-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-3-carboxamide (compound 13)

[0173] The synthesis method of compound 13 is the same as that of compound 1, except that 1c is replaced with 12c, to obtain oily compound 13 (1.0 g, yield 88%).

[0174] 1 H NMR (400MHz, CDCl3): δ7.93 (dd, J=22.2, 9.3Hz, 1H), 7.78-7.50 (m, 3H), 7.34 (d, J=5.6Hz ,1H),7.23(dt,J=15.0,7.2Hz,2H),7.16-6.93(m,2H),4.98-4.68(m,1H),4.58-4.25(m,1 H),4.06-3.26(m,4H),3.03(dd,J=36.9,7.0Hz,1H),2.86(d,J=34.6Hz,2H),2.07(d,J=10 .6Hz,2H),1.93-1.67(m,4H),1.47(ddd,J=29.3,19.2,9.0Hz,1H).ESI-MS:m / z=407[M+1] + .

[0175] Preparation Example 14. Synthesis of 7-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylisoquinoline-6-carboxamide (Compound 14)

[0176] Step 1. Synthesis of 7-fluoroisoquinoline-6-carboxynitrile (intermediate 14a)

[0177] In a 100 mL three-necked flask, 6-bromo-7-fluoroisoquinoline (2.5 g, 11.0 mmol) dissolved in DMA (25 mL) was added, followed by K3Fe(CN)6 (3.0 g, 31.3 mmol), Pd(OAc)2 (0.25 g, 10%), and Na2CO3 (4.1 g, 33.0 mmol). The mixture was heated to 125 °C and magnetically stirred for 3 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 2:1; v / v) to give solid 14a (1.8 g, 95% yield). ESI-MS: m / z = 173 [M+1] + .

[0178] Step 2. Synthesis of (1-(6-cyanoisoquinoline-7-yl)azacycloheptyl-4-yl)tert-butyl carbamate (intermediate 14b)

[0179] The synthesis of 14b was performed using the same method as 1a, except that methyl 2-fluorobenzoate was replaced with 14a and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 14b (2.0 g, 53% yield). ESI-MS: m / z = 367 [M+1] + .

[0180] Step 3.7 Synthesis of 4-(((tert-Butoxycarbonyl)amino)azaphen-1-yl)isoquinoline-6-carboxylic acid (intermediate 14c)

[0181] 14b (2.0 g, 5.5 mmol) was dissolved in EtOH (20 ml) in a 100 ml three-necked flask, followed by 2N NaOH (20 ml, 40.0 mmol). The mixture was heated to 100 °C and magnetically stirred for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and the pH was adjusted to 2 with ice-cold 1N HCl. The mixture was extracted with ethyl acetate, and the combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain solid 14c (1.5 g, 64% yield). ESI-MS: m / z = 386 [M+1] + .

[0182] Step 4. Synthesis of (1-(6-((2-fluorobenzyl)(methyl)carbamoyl)isoquinoline-7-yl)azaphen-4-yl)tert-butyl carbamate (intermediate 14d)

[0183] The synthesis method for 14d is the same as that for 1c, except that 1b is replaced with 14c and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 14d (1.3 g, yield 67%). ESI-MS: m / z = 507 [M+1] + .

[0184] Step 5. Synthesis of 7-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylisoquinoline-6-carboxamide (compound 14)

[0185] The synthesis method of compound 14 is the same as that of compound 1, except that 1c is replaced with 14d, to obtain oily compound 14 (0.9g, yield 86%).

[0186] 1H NMR (400MHz, CDCl3): δ9.13(s,1H),8.39(t,J=4.5Hz,1H),7.74(dd,J=21.8,12.6Hz,1 H),7.55(qd,J=13.0,6.1Hz,2H),7.38(dd,J=17.1,10.6Hz,2H),7.22(t,J=7.0Hz,1H) ,7.17-6.91(m,1H),5.09(dd,J=44.8,14.5Hz,1H),4.78-4.22(m,1H),3.72-2.97(m,7 H),2.82(d,J=7.4Hz,2H),2.27-1.91(m,3H),1.79-1.55(m,2H).ESI-MS:m / z=407[M+1] + .

[0187] Preparation Example 15. Synthesis of 7-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-6-carboxamide (Compound 15)

[0188] Step 1. Synthesis of 7-fluoroquinoline-6-carboxynitrile (intermediate 15a)

[0189] The synthesis method for 15a was the same as that for 14a, except that 6-bromo-7-fluoroisoquinoline was replaced with 6-bromo-7-fluoroquinoline, yielding 15a as a white solid (1.4 g, 95% yield). ESI-MS: m / z = 173 [M+1] + .

[0190] Step 2. Synthesis of (1-(6-cyanoquinolin-7-yl)azacycloheptane-4-yl)tert-butyl carbamate (intermediate 15b)

[0191] The synthesis of 15b was performed using the same method as 1a, except that methyl 2-fluorobenzoate was replaced with 15a and 4-tert-butoxycarbonylaminopiperidine was replaced with 4-tert-butoxycarbonyl-1H-azapyridine, yielding oily 15b (1.6 g, 53% yield). ESI-MS: m / z = 367 [M+1] + .

[0192] Step 3. Synthesis of 7-(4-(((tert-Butoxycarbonyl)amino)azaphen-1-yl)isoquinoline-6-carboxylic acid (intermediate 15c)

[0193] The synthesis method for 15c is the same as that for 14c, except that 14b is replaced with 15b, yielding solid 15c (1.1 g, yield 64%). ESI-MS: m / z = 386 [M+1] + .

[0194] Step 4. Synthesis of tert-butyl carbamate (intermediate 15d)

[0195] The synthesis method for 15d is the same as that for 1c, except that 1b is replaced with 15c and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding oily 15d (1.0 g, yield 67%). ESI-MS: m / z = 507 [M+1] + .

[0196] Step 5. Synthesis of 7-(4-aminoachaplon-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-6-carboxamide (compound 15)

[0197] The synthesis method of compound 15 is the same as that of compound 1, except that 1c is replaced with 15d, to obtain oily compound 15 (0.7g, yield 86%).

[0198] 1 H NMR (400MHz, CDCl3): δ8.82 (d, J=2.6Hz, 1H), 8.02 (dd, J=16.1, 8.1Hz, 1H), 7.72 (dd, J=22.9, 11.9Hz,1H),7.66-7.44(m,2H),7.31(d,J=12.6Hz,1H),7.28-7.17(m,2H),7.18-6.90(m,1H), 5.05(dd,J=41.3,14.5Hz,1H),4.67(dd,J=43.9,14.3Hz,1H),3.75-3.25(m,4H),3.23-2.98(m ,2H),2.82(d,J=15.9Hz,2H),2.07(d,J=8.2Hz,3H),1.85-1.52(m,3H).ESI-MS:m / z=407[M+1] + .

[0199] Preparation Example 16. Synthesis of 3-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 16)

[0200] Step 1. Synthesis of 3-bromonaphthalene-2-carboxylic acid (intermediate 16a)

[0201] In a 500 mL three-necked flask, 23 g (123.0 mmol) of 3-amino-2-carboxylic acid, 20 mL of 33% HBr, and 100 mL of AcOH were added. The mixture was cooled to 0 °C, and 12.7 g (184.5 mmol) of NaNO2 was added in portions. The mixture was stirred at 0 °C for 3 h, followed by the addition of 26.4 g (184.5 mmol) of CuBr in portions. The mixture was then heated to room temperature and then to 70 °C for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to give a white solid, 16a (31 g, yield: 100%). ESI-MS: m / z = 252 [M+1] + .

[0202] Step 2. Synthesis of ethyl 3-bromonaphthalene-2-carboxylate (intermediate 16b)

[0203] 16b (30 g, 120.0 mmol) was dissolved in DCM (300 ml) in a 500 ml three-necked flask, followed by CDI (29 g, 180.0 mmol) and EtOH (30 ml). The reaction was carried out at room temperature for 16 h. After the reaction was complete, the solvent was removed by rotary evaporation. The residue was diluted with water and extracted with ethyl acetate. The organic phases were combined, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 10:1, v / v) to give a white solid 16b (31 g, yield: 93%). ESI-MS: m / z = 280 [M+1] + .

[0204] Step 3. Synthesis of ethyl 3-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 16c)

[0205] In a 500 mL three-necked flask, 16C (30.0 g, 108.0 mmol) dissolved in toluene (300 mL) and 4-tert-butoxycarbonylaminopiperidine (32.2 g, 161.0 mmol) were added, along with Pd₂(DBA)₃ (3.0 g, 10%), Ruphos (3.0 g, 10%), and Cs₂CO₃ (106.0 g, 324.0 mmol). The air was replaced three times with N₂, and the mixture was heated to 110 °C for 16 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by column chromatography (PE:EA = 4:1, v / v) to give a white solid 16C (31 g, yield: 72%). ESI-MS: m / z = 399 [M+1] + .

[0206] Step 4. Synthesis of 3-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 16d)

[0207] 16c (31 g, 77.7 mmol) was dissolved in EtOH (300 ml) in a 500 ml three-necked flask, followed by the addition of 2N NaOH (50 ml, 100.0 mmol). The mixture was heated to 60 °C and reacted for 3 h. After the reaction was complete, the mixture was cooled to room temperature, and the solvent was removed by rotary evaporation. The residue was diluted with water, and the pH was adjusted to 2 with ice-cold 1N HCl. The mixture was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain solid 16d (29.0 g, yield: 100%). ESI-MS: m / z = 371 [M+1] + .

[0208] Step 5. Synthesis of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 16e)

[0209] 16d (29 g, 78.4 mmol) dissolved in DMF (150 ml) was added to a 500 ml three-necked flask. N-methyl-2-fluorobenzylamine (16.4 g, 117.6 mmol) and HATU (45 g, 117.6 mmol) were added, and the mixture was reacted at room temperature for 16 h. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give a yellow oily 16e (34 g, yield: 88%). ESI-MS: m / z = 492 [M+1] + .

[0210] Step 6. Synthesis of 3-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 16)

[0211] The synthesis method of compound 16 is the same as that of compound 1, except that 1c is replaced with 16e, to obtain compound 16 (17.9 g, yield 66%), which is a white solid.

[0212] 1H NMR (400MHz, CDCl3): δ7.84-7.72(m,3H),7.69-7.60(m,1H),7.54-7.31(m,4H),7.2 7-6.92(m,2H),5.02-4.77(m,1H),4.41(dt,J=54.9,16.4Hz,1H),3.80-3.59(m,1H), 3.42-3.20(m,1H),3.18-3.00(m,2H),2.99-2.74(m,3H),2.70-2.50(m,2H),2.15-1. 90(m,2H),1.77(s,1H),1.71-1.37(m,1H),1.33-1.15(m,1H).ESI-MS:m / z=392[M+1] + .

[0213] Preparation Example 17. Synthesis of 3-(3-aminopyrrolidone-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 17)

[0214] Step 1. Synthesis of methyl 3-(3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)-2-naphthoic acid (intermediate 17a)

[0215] In a 500 mL three-necked flask, 8b (26.0 g, 83.3 mmol) dissolved in DMSO (500 mL) was added, followed by 3-(Boc-amino)pyrrolidine (66.7 g, 333 mmol), 8-hydroxyquinoline (3.63 g, 24.99 mmol), CuI (4.76 g, 24.99 mmol), and K₂CO₃ (23 g, 167 mmol). The mixture was heated to 120 °C and reacted for 6 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 6:1, v / v) to give oily 17a (5.5 g, 18% yield). ESI-MS: m / z = 371 [M+1] + .

[0216] Step 2. Synthesis of 3-(3-((tert-butoxycarbonyl)amino)pyrrolidine-1-yl)-2-naphthoic acid (intermediate 17b)

[0217] The synthesis method for 17b is the same as that for 1b, except that 1a is replaced with 17a, yielding oily 17b (4.6 g, 88% yield). ESI-MS: m / z = 357 [M+1] + .

[0218] Step 3. Synthesis of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)pyrrolidine-3-yl)tert-butyl carbamate (intermediate 17c)

[0219] 17b (2.65 g, 7.4 mmol) was dissolved in DMF (30 ml) and added to a 500 ml three-necked flask. N-methyl-2-fluorobenzylamine (1.99 g, 14.3 mmol), TEA (1.45 g, 14.30 mmol), and PyBOP (5.95 g, 11.44 mmol) were then added, and the mixture was reacted at room temperature for 3 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 2:1, v / v) to give solid 17c (2.80 g, 79% yield). ESI-MS: m / z = 478 [M+1] + .

[0220] Step 4. Synthesis of 3-(3-aminopyrrolidone-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 17)

[0221] 17c (0.33 g, 0.69 mmol) was dissolved in MeOH (10 ml) in a 500 ml three-necked flask, followed by p-toluenesulfonic acid (0.38 g, 2.0 mmol). The reaction was carried out at 60 °C for 3 h under nitrogen protection. After the reaction was completed, NaHCO3 solution was added to adjust the reaction solution to alkaline. The mixture was extracted with ethyl acetate, and the organic phases were combined. The organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was subjected to silica gel column chromatography (PE:EA = 1:2, v / v) to give compound 17 as a solid (0.18 g, yield 72%).

[0222] 1 H NMR (500MHz, CDCl3): δ7.72(s,1H),7.79-7.71(m,1H),7.66-7.62(m,2H),7 .39-7.26(m,4H),6.99(t,J=8.0Hz,1H),7.17-7.11(m,1H),5.02-4.93(m,2H ),3.72-3.66(m,1H),3.60-3.50(m,1H),3.48-3.38(m,2H),3.37-3.30(m,1H ),3.03(s,3H),2.03-1.99(m,3H),1.93-1.82(m,1H).ESI-MS:m / z=378[M+1] + .

[0223] Preparation Example 18. Synthesis of 3-(3-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 18)

[0224] Step 1. Synthesis of methyl 3-(3-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 18a)

[0225] The synthesis method for 18a is the same as that for 17a, except that the starting material 3-(Boc-amino)pyrrolidine is replaced with 3-(Boc-amino)piperidine, yielding solid 18a (0.3 g, yield 12%). ESI-MS: m / z = 385 [M+1] + .

[0226] Step 2. Synthesis of 3-(3-(tert-Butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 18b)

[0227] The synthesis method for 18b is the same as that for 1b, except that the starting material 1a is replaced with 18a, yielding solid 18b (0.3 g, yield 87%). ESI-MS: m / z = 371 [M+1] + .

[0228] Step 3. Synthesis of tert-butyl carbamate (intermediate 18c)

[0229] The synthesis method for 18c is the same as that for 17c, except that the starting material 17b is replaced with 18b, yielding oily 18c (0.2 g, 60% yield). ESI-MS: m / z = 492 [M+1] + .

[0230] Step 4. Synthesis of 3-(3-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 18)

[0231] The synthesis method of compound 18 is the same as that of compound 17, except that 17c is replaced by 18c, to obtain solid compound 18 (0.1g, yield 72%).

[0232] 1H NMR (500MHz, CDCl3): δ7.83(s,1H),7.78-7.73(m,2H),7.51-7.45(m,1H),7.39-7.24(m,4H ),7.21(td,J=8.0,1.5Hz,1H),6.96(t,1H,J=8.0,1H),5.00(d,J=14.7Hz,1H),4.86(d,J=1 4.6Hz,1H),4.48-4.43(m,1H),3.67-3.58(m,1H),3.51-3.33(m,1H),3.20-3.10(m,2H),3. 03(s,3H),1.98-1.78(m,2H),1.82-1.70(m,3H),1.69-1.54(m,1H).ESI-MS:m / z=392[M+1] + .

[0233] Preparation Example 19. Synthesis of 2-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-1-naphthamide (compound 19)

[0234] Step 1. Synthesis of 2-bromo-1-methylnaphthalene (intermediate 19a)

[0235] 1-Methylnaphthalene (5.0 g, 35.2 mmol) and H₂O (15 mL) were added to a 100 mL three-necked flask, followed by 30% H₂O₂ (2.39 g, 70.3 mmol) and 48% HBr (3.13 g, 38.7 mmol). The reaction was carried out at room temperature in the dark for 3 h. After the reaction was completed, the mixture was quenched with sodium sulfite solution, diluted with water, extracted with ethyl acetate, and the organic layers were combined. The mixture was dried over anhydrous sodium sulfate and evaporated to dryness using a rotary evaporator to obtain oil 19a (6.50 g, 84% yield). ESI-MS: m / z = 221 [M+1] + .

[0236] Step 2. Synthesis of 2-bromo-1-naphthoic acid (intermediate 19b)

[0237] In a 100 mL three-necked flask, 19a (1.00 g, 4.5 mmol) dissolved in pyridine (9 mL) and H₂O (15 mL) was added, followed by potassium permanganate (5.0 g, 31.0 mmol). The mixture was heated to 120 °C and reacted for 1 h. After the reaction was complete, the mixture was cooled to room temperature, quenched with sodium sulfite solution, and then adjusted to acidity with dilute hydrochloric acid. The solution was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give oily 19b (0.9 g, 80% yield). ESI-MS: m / z = 251 [M+1] + .

[0238] Step 3. Synthesis of 2-bromo-N-(2-fluorobenzyl)-N-methyl-1-naphthamide (intermediate 19c)

[0239] The synthesis method for 19c is the same as that for 17c, except that the starting material 17b is replaced with 19b, yielding oily 19c (0.9 g, yield 65%). ESI-MS: m / z = 372 [M+1] + .

[0240] Step 4. Synthesis of tert-butyl carbamate (19d)

[0241] In a 100 mL three-necked flask, 19c (330 mg, 0.89 mmol) dissolved in 1,4-dioxane (10 mL) was added, followed by 4-tert-butoxycarbonylaminopiperidine (160 mg, 0.80 mmol), RuPhos Pd G3 (332 mg, 0.038 mmol), and Cs₂CO₃ (520 mg, 1.60 mmol). The mixture was refluxed under nitrogen protection for 6 h. After the reaction was complete, the mixture was cooled to room temperature, diluted with water, and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give oily 19c (250 mg, 64% yield). ESI-MS: m / z = 492 [M+1] + .

[0242] Step 5. Synthesis of 2-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-1-naphthamide (compound 19)

[0243] The synthesis method of compound 19 is the same as that of compound 17, except that 17c is replaced with 19c, to obtain solid compound 19 (0.1g, yield 56%).

[0244] 1 H NMR (500MHz, CDCl3): δ8.23-8.18 (m, 2H), 7.83 (dd, J = 8.0, 1.5Hz, 1H), 7.47-7. 40(m,1H),7.39-7.32(m,2H),7.33-7.27(m,1H),7.27-7.18(m,2H),7.18-7.09 (m,1H),4.99(s,2H),3.47-3.41(m,2H),3.28-3.11(m,5H),3.10-2.98(m,1H), 2.92-2.78(m,2H),2.16-2.05(m,2H),1.92-1.78(m,2H).ESI-MS:m / z=392[M+1] + .

[0245] Preparation Example 20. Synthesis of 1-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 20)

[0246] Step 1. Synthesis of methyl 1-(4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 20a)

[0247] The synthesis method for 20a is the same as that for 17a, except that starting material 8b is replaced with methyl 1-iodo-2-naphthoate, yielding oily 20a (0.4 g, yield 15%). ESI-MS: m / z = 385 [M+1] + .

[0248] Step 2.1 Synthesis of 4-((tert-butoxycarbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 20b)

[0249] The synthesis method for 20b is the same as that for 1b, except that raw material 1a is replaced with 20a, yielding solid 20b (0.4 g, yield 91%). ESI-MS: m / z = 371 [M+1] + .

[0250] Step 3. Synthesis of (1-(2-((2-fluorobenzyl)(methyl)carbamoyl)naphth-1-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 20c)

[0251] The synthesis method for 20c is the same as that for 17c, except that the starting material 17b is replaced with 20b, yielding oily 20c (0.4 g, yield 73%). ESI-MS: m / z = 492 [M+1] + .

[0252] Step 4. Synthesis of 1-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 20)

[0253] The synthesis method of compound 20 is the same as that of compound 17, except that 17c is replaced with 20c, to obtain oily compound 20 (0.2g, yield 53%). 1 H NMR (500MHz, CDCl3): δ8.32 (dd, J=8.0, 1.5Hz, 1H), 7.82 (dd, J=8.5, 1.5Hz, 1H), 7.75 (d, J=8.5 Hz,1H),7.71-7.62(m,2H),7.55-7.49(m,1H),7.38-7.27(m,2H),7.25-7.18(m,1H),7.17-7.1 0(m,1H),5.00(d,J=14.7Hz,1H),4.74(d,J=14.6Hz,1H),3.65-3.44(m,2H),3.38-3.24(m,2H) ,3.14(s,3H),3.16-3.06(m,3H),2.09-1.87(m,2H),1.81-1.61(m,2H).ESI-MS:m / z=392[M+1] + .

[0254] Preparation Example 21. Synthesis of 3-(3-(aminomethyl)azacyclobut-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthylcarboxamide (Compound 21)

[0255] Step 1. Synthesis of methyl 3-(3-(((tert-Butoxycarbonyl)amino)methyl)azacyclobut-1-yl)-2-naphthoate (intermediate 21a)

[0256] The synthesis method for 21a is the same as that for 17a, except that the starting material 3-(Boc-amino)pyrrolidine is replaced with 3-Boc-aminomethylazinecyclobutane, yielding oily 21a (0.5 g, yield 21%). ESI-MS: m / z = 371 [M+1] + .

[0257] Step 2.3 Synthesis of 3-(((tert-Butoxycarbonyl)amino)methyl)azacyclobut-1-yl)-2-naphthoic acid (intermediate 21b)

[0258] The synthesis method of 21b is the same as that of 1b, except that the starting material 1a is replaced with 21a, yielding solid 21b (0.4 g, yield 85%). ESI-MS: m / z = 357 [M+1] + .

[0259] Step 3. Synthesis of ((1-(3-(((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)azacyclobut-3-yl)methyl)tert-butyl carbamate (intermediate 21c)

[0260] The synthesis method for 21c is the same as that for 17c, except that the starting material 17b is replaced with 21b, yielding oily 21c (0.4 g, 75% yield). ESI-MS: m / z = 478 [M+1] + .

[0261] Step 4. Synthesis of 3-(3-(aminomethyl)azacyclobut-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthylcarboxamide (compound 21)

[0262] The synthesis method of compound 21 is the same as that of compound 1, except that 1c is replaced with 21c, to obtain oily compound 21 (0.2g, yield 70%).

[0263] 1 H NMR (500MHz, CDCl3): δ72-7.66(m,2H),7.57(t,J=8.0Hz,1H),7.43-7.36(m,2H ),7.27-7.15(m,3H),7.24-7.17(m,1H),7.17-7.07(m,1H),5.05(d,J=14.7Hz,1 H), 4.66 (d, J = 14.6Hz, 1H), 4.04-3.82 (m, 2H), 3.70-3.58 (m, 2H), 3.07 (s, 3H), 2.93-2.74(m,2H),2.28-2.07(m,1H),2.04-1.86(m,2H).ESI-MS:m / z=378[M+1] + .

[0264] Preparation Example 22. Synthesis of 3-(6-amino-3-azabicyclo[3.1.1]hept-3-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 22)

[0265] Step 1. Synthesis of ethyl 3-(6-((tert-butoxycarbonyl)amino)-3-azabicyclo[3.1.1]heptane-3-yl)-2-naphthoic acid (intermediate 22a)

[0266] The synthesis method for 22a is the same as that for 19d, except that starting material 19c is replaced with 16b, and 4-tert-butoxycarbonylaminopiperidine is replaced with 3-azabicyclo[3.1.1]heptane-6-ylcarbamate tert-butyl ester, yielding solid 22a (0.4 g, yield 7%). ESI-MS: m / z = 411 [M+1] + .

[0267] Step 2. Synthesis of 3-(6-((tert-butoxycarbonyl)amino)-3-azabicyclo[3.1.1]heptane-3-yl)-2-naphthoic acid (intermediate 22b)

[0268] The synthesis method of 22b is the same as that of 1b, except that the starting material 1a is replaced with 22a, yielding solid 22b (0.3 g, yield 87%). ESI-MS: m / z = 383 [M+1] + .

[0269] Step 3. Synthesis of tert-butyl carbamate (intermediate 22c) of (3-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)-3-azabicyclo[3.1.1]heptane-6-yl)carbamate

[0270] The synthesis method for 22c is the same as that for 17c, except that the starting material 17b is replaced with 22b, yielding solid 22c (0.3 g, yield 61%). ESI-MS: m / z = 504 [M+1] + .

[0271] Step 4. Synthesis of 3-(6-amino-3-azabicyclo[3.1.1]hept-3-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 22)

[0272] The synthesis method of compound 22 is the same as that of compound 17, except that 17c is replaced with 22c, to obtain solid 22 (0.1g, yield 68%).

[0273] 1H NMR (500MHz, CDCl3): δ7.93 (d, J = 1.5Hz, 1H), 7.78-7.74 (m, 2H), 7.50-7.45 (m, 1H), 7.40 -7.26(m,4H),7.24-7.18(m,1H),7.16-7.11(m,1H),4.94-4.83(m,2H),3.98-3.67(m,2H),3.67-3.53(m ,2H),3.16-3.01(m,4H),2.59-2.43(m,2H),2.23-2.03(m,2H),1.96-1.83(m,2H).ESI-MS:m / z=404[M+1] + .

[0274] Preparation Example 23. Synthesis of 3-(6-amino-2-azaspirocyclic[3.3]heptane-2-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 23)

[0275] Step 1. Synthesis of methyl 3-(6-(tert-butoxycarbonyl)amino-2-azaspiro[3.3]hept-2-yl)-2-naphthoic acid (intermediate 23a)

[0276] The synthesis method for 23a is the same as that for 17a, except that the starting material 3-(Boc-amino)pyrrolidine is replaced with N-2-azaspiro[3.3]hept-6-ylcarbamate tert-butyl ester, yielding solid 23a (0.6 g, yield 23%). ESI-MS: m / z = 397 [M+1] + .

[0277] Step 2. Synthesis of 3-(6-(tert-Butoxycarbonyl)amino-2-azaspiro[3.3]hept-2-yl)-2-naphthoic acid (intermediate 23b)

[0278] The synthesis method for 23b is the same as that for 1b, except that raw material 1a is replaced with 23a, yielding solid 23b (0.5g, yield 93%). ESI-MS: m / z = 383 [M+1] + .

[0279] Step 3. Synthesis of tert-butyl carbamate (intermediate 23c) of (2-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)-2-azaspiro[3.3]heptane-6-yl)carbamate

[0280] The synthesis method for 23c is the same as that for 17c, except that the starting material 17b is replaced with 23b, yielding solid 23c (0.5g, yield 76%). ESI-MS: m / z = 504 [M+1] +.

[0281] Step 4. Synthesis of 3-(6-amino-2-azaspirocyclic[3.3]heptane-2-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 23)

[0282] The synthesis method of compound 23 is the same as that of compound 1, except that 1c is replaced with 23c, to obtain solid 23 (0.3g, yield 79%).

[0283] 1 H NMR (500MHz, CDCl3): δ7.85 (d, J = 1.5Hz, 1H), 7.78-7.72 (m, 2H), 7.62-7.54 (m, 1H), 7.38-7.26 (m, 4H), 7.24-7.18 (m, 1H), 7.17-7.10 (m, 1H), 5. 03(d,J=14.7Hz,1H), 4.74(d,J=14.6Hz,1H), 378-3.61(m,2H), 3.40-3. 23(m,2H),3.16-2.96(m,6H),1.96-1.74(m,4H).ESI-MS:m / z=404[M+1] + .

[0284] Preparation Example 24. Synthesis of 3-(3-aminoazacyclobutane-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 24)

[0285] Step 1. Synthesis of methyl 3-(3-((tert-butoxycarbonyl)amino)azacyclobutane-1-yl)-2-naphthoate (intermediate 24a)

[0286] The synthesis method for 24a is the same as that for 17a, except that the starting material 3-(Boc-amino)pyrrolidine is replaced with 3-(Boc-amino)azacyclobutane, yielding oily 24a (0.4 g, yield 17%). ESI-MS: m / z = 357 [M+1] + .

[0287] Step 2. Synthesis of 3-(3-((tert-butoxycarbonyl)amino)azacyclobutane-1-yl)-2-naphthoic acid (intermediate 24b)

[0288] The synthesis method of 24b is the same as that of 1b, except that the starting material 1a is replaced with 24a, yielding oily 24b (0.3g, yield 92%). ESI-MS: m / z = 343 [M+1] + .

[0289] Step 3. Synthesis of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)azacyclobutane-3-yl)tert-butyl carbamate (intermediate 24c)

[0290] The synthesis method for 24c is the same as that for 17c, except that the starting material 17b is replaced with 24b, yielding oily 24c (0.3g, yield 82%). ESI-MS: m / z = 464 [M+1] + .

[0291] Step 4. Synthesis of 3-(3-aminoazacyclobutane-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 24)

[0292] The synthesis method of compound 24 is the same as that of compound 1, except that 1c is replaced with 24c, to obtain oily compound 24 (0.1g, yield 21%).

[0293] 1 H NMR (500MHz, CDCl3): δ7.71 (d, J = 1.5Hz, 1H), 7.77-7.66 (m, 2H), 7.44-7.38 (m, 2H), 7.28-7.20 (m, 3H), 7.18-7.10 (m, 2H), 5.10 (d, J = 14.7Hz, 1 H), 4.64 (d, J = 14.6Hz, 1H), 4.30 (t, J = 8.0Hz, 1H), 4.00-3.85 (m, 2H), 3.72-3.59 (m, 2H), 3.58-3.45 (m, 2H), 3.09 (s, 3H). ESI-MS: m / z = 364 [M+1] + .

[0294] Preparation Example 25. Synthesis of 3-(4-aminopiperidin-1-yl)-4-fluoro-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 25)

[0295] Step 1. Synthesis of ethyl 3-(4-(tert-Butoxycarbonyl)amino)piperidin-1-yl)-4-fluoro-2-naphthoic acid (intermediate 25a)

[0296] 8c (150 g, 0.4 mmol) dissolved in DMF (5 ml) was added to a 100 ml three-necked flask. The mixture was cooled to 0–5 °C, and 1-chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane di(tetrafluoroborate) salt (138 mg, 0.4 mmol) was added. The mixture was magnetically stirred at room temperature for 2 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic layers were combined. The layers were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 4:1, v / v) to give solid 25a (60 mg, yield 38%). ESI-MS: m / z = 417 [M+1] + .

[0297] Step 2. Synthesis of 3-(4-(tert-Butoxycarbonyl)amino)piperidin-1-yl)-4-fluoro-2-naphthoic acid (intermediate 25b)

[0298] The synthesis method for 25b is the same as that for 1b, except that 1a is replaced with 25a, yielding solid 25b (0.05 g, 90% yield). ESI-MS: m / z = 389 [M+1] + .

[0299] Step 3. Synthesis of tert-butyl carbamate (intermediate 25c)

[0300] The synthesis method for 25c is the same as that for 1c, except that 1b is replaced with 25b and N-methylbenzylamine is replaced with N-methyl-2-fluorobenzylamine, yielding solid 25c (0.05 g, 75% yield). ESI-MS: m / z = 510 [M+1] + .

[0301] Step 4. Synthesis of 3-(4-aminopiperidin-1-yl)-4-fluoro-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 25)

[0302] The synthesis method of compound 25 is the same as that of compound 1, except that 1c is replaced with 25c, to obtain solid compound 25 (0.03g, yield 82%).

[0303] 1H NMR (500MHz, CDCl3): δ8.05-7.98(m,2H),7.78(d,J=8.0Hz,1H),7.65(td,J=8. 0,1.5Hz,1H),7.38-7.27(m,3H),7.24-7.18(m,1H),7.17-7.11(m,1H),4.97(d ,J=14.6Hz,1H),4.84(d,J=14.6Hz,1H),3.66-3.52(m,2H),3.46-3.31(m,2H), 3.12-3.00(m,4H),2.08-1.92(m,2H),1.79-1.62(m,4H).ESI-MS:m / z=410[M+1] + .

[0304] Preparation Example 26. Synthesis of 3-(4-amino-4-methylpiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 26)

[0305] Step 1. Synthesis of ethyl 3-(4-(tert-Butoxycarbonyl)amino)piperidin-1-yl)-4-fluoro-2-naphthoic acid (intermediate 26a)

[0306] The synthesis method for 26a is the same as that for 16c, except that the starting material 4-tert-butoxycarbonylaminopiperidine is replaced with (4-methylpiperidin-4-yl)carbamate tert-butyl ester, yielding oily 26a (1.6 g, yield 54%). ESI-MS: m / z = 413 [M+1] + .

[0307] Step 2. Synthesis of 3-(4-(tert-Butoxycarbonyl)amino)piperidin-1-yl)-4-fluoro-2-naphthoic acid (intermediate 26b)

[0308] The synthesis method for 26b is the same as that for 1b, except that raw material 1a is replaced with 26a, yielding solid 26b (1.3g, 90% yield). ESI-MS: m / z = 385 [M+1] + .

[0309] Step 3. Synthesis of tert-butyl carbamate (intermediate 26c) of (1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)-4-methylpiperidin-4-yl)carbamate

[0310] The synthesis method for 26c is the same as that for 16e, except that the starting material 16d is replaced with 26b, yielding oily 26c (1.7 g, 96% yield). ESI-MS: m / z = 506 [M+1] + .

[0311] Step 4. Synthesis of 3-(4-amino-4-methylpiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 26)

[0312] The synthesis method of compound 26 is the same as that of compound 1, except that 1c is replaced with 26c, to obtain oily compound 26 (0.6g, yield 44%).

[0313] 1 H NMR (400MHz, DMSO): δ7.88-7.76(m,2H),7.50-7.42(m,2H),7.41-7.32(m,2H) ,7.29-7.19(m,2H),7.16-7.01(m,2H),4.97-4.52(m,1H),4.32(s,1H),3.23- 3.08(m,2H),3.04-2.96(m,1H),2.92(s,1H),2.77(dd,J=22.5,9.3Hz,1H),2. 68(s,2H),1.90-1.48(m,4H),1.30(s,1H),1.22(s,2H).ESI-MS:m / z=406[M+1] + .

[0314] Preparation Example 27. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2-fluoropyridin-3-yl)methyl)-N-methyl-2-naphthamide (compound 27)

[0315] Step 1. Synthesis of 1-(2-fluoropyridin-3-yl)-N-methylmethylamine (intermediate 27a): 2-fluoro-3-pyridinecarboxaldehyde (2 g, 16 mmol) and methylamine (8.28 g, 30% MeOH solution, 80 mmol) dissolved in methanol (20 mL) were added to a 100 mL three-necked flask. AcOH (4.8 g) was added, and the mixture was magnetically stirred at room temperature for 2 h. Then, NaBH3CN (2 g, 32 mmol) was added, and the mixture was magnetically stirred at room temperature for 12 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Purification was performed by silica gel column chromatography (DCM:MeOH = 10:1, v / v) to give a yellow oily 27a (1.5 g, yield 67%). ESI-MS: m / z = 140 [M+1] + .

[0316] Step 2. Synthesis of tert-butyl carbamate (intermediate 27b)

[0317] The synthesis method for 27b is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 27a, yielding oily 27b (5.0 g, 94% yield). ESI-MS: m / z = 493 [M+1] + .

[0318] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2-fluoropyridin-3-yl)methyl)-N-methyl-2-naphthamide (compound 27)

[0319] The synthesis method of compound 27 is the same as that of compound 1, except that 1c is replaced with 27b, to obtain solid compound 27 (2.0 g, yield 50%).

[0320] 1 H NMR (400MHz, DMSO): δ8.18(d,J=4.8Hz,1H),7.98-7.89(m,1H),7.89-7.70(m,5 H),7.53-7.22(m,4H),5.08-4.97(m,1H),4.47-4.21(m,1H),3.49(t,J=14.0Hz, 1H),3.13(dt,J=11.4,9.4Hz,1H),3.08-2.99(m,1H),2.96(s,1H),2.73(s,2H), 2.65-2.50(m,2H),2.08-1.82(m,2H),1.75-1.43(m,2H).ESI-MS:m / z=393[M+1] + .

[0321] Preparation Example 28. Synthesis of 3-(4-amino-3,3-difluoropiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (Compound 28)

[0322] Step 1. Synthesis of ethyl 3-(4-((tert-Butoxycarbonyl)amino)-3,3-difluoropiperidin-1-yl)-2-naphthoic acid (intermediate 28a)

[0323] The synthesis method for 28a is the same as that for 16c, except that the starting material 4-tert-butoxycarbonylaminopiperidine is replaced with (3,3-difluoropiperidin-4-yl)carbamate tert-butyl ester, yielding oily 28a (2.8 g, 90% yield). ESI-MS: m / z = 435 [M+1] + .

[0324] Step 2. Synthesis of 3-(4-((tert-Butoxycarbonyl)amino)-3,3-difluoropiperidin-1-yl)-2-naphthoic acid (intermediate 28b)

[0325] The synthesis method of 28b is the same as that of 1b, except that the starting material 1a is replaced with 28a, yielding oily 28b (2.5 g, 94% yield). ESI-MS: m / z = 407 [M+1] + .

[0326] Step 3. Synthesis of tert-butyl carbamate (intermediate 28c) of (3,3-difluoro-1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 28c)

[0327] The synthesis method for 28c is the same as that for 16e, except that the starting material 16d is replaced with 28b, yielding oily 28c (2.0 g, yield 63%). ESI-MS: m / z = 528 [M+1] + .

[0328] Step 4. Synthesis of 3-(4-amino-3,3-difluoropiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 28)

[0329] The synthesis method of compound 28 is the same as that of compound 1, except that 1c is replaced with 28c, to obtain solid compound 28 (1.4g, yield 84%).

[0330] 1 H NMR (400MHz, DMSO): δ7.97-7.74(m,3H),7.57-7.28(m,4H),7.29-7.17(m,2H),7.14-7.00(m,1H),5.16-4.51(m,1H),4.44-4.13( m,1H),3.55-3.32(m,2H),3.22-2.90(m,2H),2.91(d,J=7.5Hz,1H),2.72-2.55(m,2H),1.97-1.42(m,4H).ESI-MS:m / z=428[M+1] + .

[0331] Preparation Example 29. Synthesis of 3-(4-(aminomethyl)piperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 29)

[0332] Step 1. Synthesis of ethyl 3-(4-((((tert-Butoxycarbonyl)amino)methyl)piperidin-1-yl)-2-naphthoic acid (intermediate 29a)

[0333] The synthesis method for 29a is the same as that for 16c, except that the starting material 4-tert-butoxycarbonylaminopiperidine is replaced with 4-Boc-aminomethylpiperidine, yielding oily 29a (2.6 g, 89% yield). ESI-MS: m / z = 413 [M+1] + .

[0334] Step 2. Synthesis of 3-(4-((((tert-Butoxycarbonyl)amino)methyl)piperidin-1-yl)-2-naphthoic acid (intermediate 29b)

[0335] The synthesis method for 29b is the same as that for 1b, except that raw material 1a is replaced with 29a, yielding solid 29b (2.0 g, yield 81%). ESI-MS: m / z = 385 [M+1] + .

[0336] Step 3. Synthesis of tert-butyl carbamate (intermediate 29c) of 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidin-4-yl)methyl)carbamate

[0337] The synthesis method for 29c is the same as that for 16e, except that the starting material 16d is replaced with 29b, yielding oily 29c (2.4 g, 91% yield). ESI-MS: m / z = 506 [M+1] + .

[0338] Step 4. Synthesis of 3-(4-(aminomethyl)piperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 29)

[0339] The synthesis method of compound 29 is the same as that of compound 1, except that 1c is replaced with 29c, to obtain solid 29 (1.2g, yield 64%).

[0340] 1 H NMR (400MHz, DMSO): δ7.98-7.71(m,3H),7.67-7.31(m,5H),7.27-6.98(m,2H),4. 94-4.57(m,1H),4.47-4.17(m,1H),3.70-3.37(m,1H),3.37(s,3H),3.05–2.81(m ,2H),2.73-2.64(m,2H),2.57-2.48(m,1H),2.41(d,J=6.6Hz,1H),2.31(ddd,J=7 .9,5.9,5.5Hz,1H),1.92-1.67(m,1H),1.59-0.75(m,4H).ESI-MS:m / z=406[M+1] + .

[0341] Preparation Example 30. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-4-yl)methyl)-N-methyl-2-naphthamide (compound 30)

[0342] Step 1. Synthesis of 1-(3-fluoropyridin-4-yl)-N-methylmethylamine (intermediate 30a)

[0343] The synthesis method for 30a is the same as that for 27a, except that the starting material 2-fluoro-3-pyridinecarboxaldehyde is replaced with 4-fluoro-3-formylpyridine, yielding oily 30a (1.4 g, yield 63%). ESI-MS: m / z = 140 [M+1] + .

[0344] Step 2. Synthesis of tert-butyl carbamate (intermediate 30b)

[0345] The synthesis method of 30b is the same as that of 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 30a, yielding oily 30b (2.6 g, yield 52%). ESI-MS: m / z = 493 [M+1] + .

[0346] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-4-yl)methyl)-N-methyl-2-naphthamide (compound 30)

[0347] The synthesis method of compound 30 is the same as that of compound 1, except that 1c is replaced with 30b, to obtain solid compound 30 (1.8g, yield 89%).

[0348] 1 H NMR (500MHz, CDCl3): δ8.53-8.46(m,2H),7.90-7.84(m,2H),7.79-7.72(m,1H),7.51-7.30(m,4H),5.05(d,J=15.4Hz,1H),4.72(d,J =15.4Hz,1H),3.52-3.43(m,2H),3.33-3.26(m,2H),3.11-2.99(m,4H),2.05-1.95(m,2H),1.78-1.64(m,4H).ESI-MS:m / z=393[M+1] + .

[0349] Preparation Example 31. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-2-yl)methyl)-N-methyl-2-naphthamide (compound 31)

[0350] Step 1. Synthesis of 1-(3-fluoropyridin-2-yl)-N-methylmethylamine (intermediate 31a)

[0351] The synthesis method for 31a is the same as that for 27a, except that the starting material 2-fluoro-3-pyridinecarboxaldehyde is replaced with 3-fluoropyridine-2-carboxaldehyde, yielding oily 31a (1.4 g, yield 62%). ESI-MS: m / z = 140 [M+1] + .

[0352] Step 2. Synthesis of tert-butyl carbamate (intermediate 31b)

[0353] The synthesis method for 31b is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 31a, yielding solid 31b (3.7 g, 75% yield). ESI-MS: m / z = 493 [M+1] + .

[0354] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-2-yl)methyl)-N-methyl-2-naphthamide (compound 31)

[0355] The synthesis method of compound 31 is the same as that of compound 1, except that 1c is replaced with 31b, to obtain solid compound 31 (1.5g, yield 52%).

[0356] 1 H NMR (500MHz, CDCl3): δ8.36 (dd, J=8.0, 2.0Hz, 1H), 7.99 (d, J=2.0Hz, 1H), 7.78 -7.73(m,2H),7.53-7.45(m,2H),7.44-7.39(m,2H),7.35-7.30(m,1H),5.26(d ,J=15.6Hz,1H),4.75(d,J=15.6Hz,1H),3.57-3.38(m,2H),3.38-3.24(m,2H), 3.16-2.99(m,4H),2.07-1.95(m,2H),1.79-1.61(m,4H).ESI-MS:m / z=393[M+1] + .

[0357] Preparation Example 32. Synthesis of 3-(4-aminopiperidin-1-yl)-N-cyclopropyl-N-((3-fluoropyridin-2-yl)methyl)-2-naphthamide (compound 32)

[0358] Step 1. Synthesis of N-(3-fluoropyridin-2-yl)methyl)cyclopropylamine (intermediate 32a)

[0359] The synthesis method for 32a is the same as that for 27a, except that the starting material 2-fluoro-3-pyridinecarboxaldehyde is replaced with 3-fluoropyridine-2-carboxaldehyde, and methylamine is replaced with cyclopropylamine, yielding oily 32a (1.9 g, yield 71%). ESI-MS: m / z = 167 [M+1] + .

[0360] Step 2. Synthesis of tert-butyl carbamate (intermediate 32b)

[0361] The synthesis method for 32b is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 32a, yielding oily 32b (2.8 g, yield 48%). ESI-MS: m / z = 519 [M+1] + .

[0362] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-cyclopropyl-N-((3-fluoropyridin-2-yl)methyl)-2-naphthamide (compound 32)

[0363] The synthesis method of compound 32 is the same as that of compound 1, except that 1c is replaced with 32b, to obtain solid compound 32 (2.0 g, yield 86%).

[0364] 1 H NMR (500MHz, CDCl3): δ8.36 (dd, J=8.0, 2.2Hz, 1H), 8.09 (d, J=2.5Hz, 1H), 7.92-7.8 4(m,1H),7.78-7.73(m,1H),7.53-7.39(m,5H),4.98(d,J=15.6Hz,1H),4.78(d,J=1 5.6Hz,1H),3.53-3.43(m,2H),3.34-3.26(m,4H),3.12-3.04(m,2H),2.14-1.90(m, 2H),1.82-1.69(m,2H),1.65-1.58(m,2H),1.49-1.30(m,2H).ESI-MS:m / z=419[M+1] + .

[0365] Preparation Example 33. Synthesis of N-((3-(4-aminopiperidin-1-yl)naphth-2-yl)methyl)-2-fluoro-N-methylbenzamide (Compound 33)

[0366] Step 1. Synthesis of tert-butyl carbamate (1-(3-(hydroxymethyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 33a)

[0367] 16b (600 mg, 1.56 mmol) was dissolved in anhydrous THF (15 mL) in a 100 mL three-necked flask. The mixture was cooled to 0–5 °C, and LiAlH4 (72 mg, 1.87 mmol) was added. The mixture was kept at this temperature and magnetically stirred for 3 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give oily 33a (350 mg, yield 63%). ESI-MS: m / z = 357 [M+1] + .

[0368] Step 2. Synthesis of (1-(3-formylnaphthalene-2-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 33b)

[0369] 33a (150 mg, 0.42 mmol) dissolved in DCM (30 ml) was added to a 100 ml three-necked flask. The mixture was cooled to 0–5 °C, and Dess-Martin (268 mg, 0.63 mmol) was added in portions. After the addition was complete, the mixture was brought to room temperature and magnetically stirred for 2 h. After the reaction was complete, the mixture was diluted with water, extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 4:1, v / v) to give oily 33b (118 mg, 79% yield). ESI-MS: m / z = 355 [M+1] + .

[0370] Step 3. Synthesis of tert-butyl carbamate (intermediate 33c)

[0371] 33b (118 mg, 0.3 mmol) dissolved in MeOH (10 mL) was added to a 100 mL three-necked flask. Methylamine hydrochloride (20 mg, 0.3 mmol) and K₂CO₃ (14 mg, 1.0 mmol) were added, and the mixture was magnetically stirred at room temperature for 2 h. The mixture was then cooled to 0–5 °C, and sodium borohydride (14 mg, 0.3 mmol) was added in portions. After the addition was complete, the mixture was kept at room temperature for 1 h, and then allowed to react for another 2 h. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (DCM:MeOH:TEA = 30:1:1, v / v) to give oily 33c (79 mg, 71% yield). ESI-MS: m / z = 370 [M+1] + .

[0372] Step 4. Synthesis of tert-butyl carbamate (intermediate 33d)

[0373] The synthesis method for 33d is the same as that for 17c, except that 17b is replaced with 33c, N-methyl-2-fluorobenzylamine is replaced with o-fluorobenzoic acid, and 17b is replaced with 33c, yielding oily 33d (0.074 g, 70% yield). ESI-MS: m / z = 492 [M+1] + .

[0374] Step 5. Synthesis of N-((3-(4-aminopiperidin-1-yl)naphth-2-yl)methyl)-2-fluoro-N-methylbenzamide (compound 33)

[0375] The synthesis method of compound 33 is the same as that of compound 17, except that 17c is replaced with 33c, to obtain oily compound 33 (0.046g, yield 78%).

[0376] 1 H NMR (500MHz, CDCl3): δ7.86-7.79(m,1H),7.78-7.71(m,1H),7.71-7.64(m,1H),7.53-7.44(m,2H),7.36-7.21(m,4H),7.09-7.06(m,1 H),4.62(s,2H),3.48-3.33(m,2H),3.32-3.21(m,2H),3.18-2.98(m,4H),2.07-1.88(m,2H),1.82-1.62(m,4H).ESI-MS: m / z=392[M+1] + .

[0377] Preparation Example 34. Synthesis of N-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-2-(2-fluorophenyl)acetamide (compound 34)

[0378] Step 1. Synthesis of (1-(3-aminonaphthyl-2-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 34a)

[0379] 16a (400 mg, 1.08 mmol) dissolved in DMF (30 mL) was added to a 100 mL three-necked flask, followed by slow addition of DPPA (448 mg, 1.62 mmol). The mixture was magnetically stirred at room temperature for 3 h. Then, 12 mL of water was added, and the mixture was heated to 90 °C for 1 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and the organic layer was washed with sodium bicarbonate solution and saturated sodium chloride solution, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The purified organic layer was then purified by silica gel column chromatography (PE:EA = 6:1, v / v) to give an oily 34a (100 mg, 27% yield). ESI-MS: m / z = 342 [M+1] + .

[0380] Step 2. Synthesis of (1-(3-(2-(2-fluorophenyl)acetamido)naphth-2-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 34b)

[0381] The synthesis method for 34b is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 2-(2-fluorophenyl)acetic acid, and 16d is replaced with 34a, yielding oily 34b (0.1 g, 80% yield). ESI-MS: m / z = 478 [M+1] + .

[0382] Step 3. Synthesis of N-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-2-(2-fluorophenyl)acetamide (compound 34)

[0383] The synthesis method of compound 34 is the same as that of compound 17, except that 17c is replaced with 34b, to obtain solid compound 34 (0.07g, yield 85%).

[0384] 1H NMR (400MHz, CDCl3): δ8.91(s,1H),8.85(s,1H),7.84-7.76(m,1H),7.67(dd,J=8.0,1.5Hz,1H),7.50(s,1H),7.48-7.34(m,4H),7.27-7.17(m,2 H),3.87(d,J=1.5Hz,2H),3.08-2.93(m,3H),2.79-2.61(m,2H),2.02-1 .88(m,2H),1.51-1.35(m,2H),1.34-1.21(m,2H).ESI-MS:m / z=378[M+1] + .

[0385] Preparation Example 35. Synthesis of 2-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-N-(2-fluorophenyl)acetamide (compound 35)

[0386] Step 1. Synthesis of methyl 3-(4-aminopiperidin-1-yl)-2-naphthoic acid ester (intermediate 35a)

[0387] 16c (2.0 g, 5.2 mmol) was dissolved in DCM (20 mL) and added to a 100 mL three-necked flask. TFA (3.0 g, 26.0 mmol) was added, and the mixture was magnetically stirred at room temperature for 2 h. After the reaction was complete, a saturated sodium bicarbonate solution was added, and the mixture was extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain an oily 35a (1.4 g, 95% yield). ESI-MS: m / z = 285 [M+1] + .

[0388] Step 2. Synthesis of methyl 3-(4-((benzyloxy)carbonyl)amino)piperidin-1-yl)-2-naphthoic acid (intermediate 35b)

[0389] In a 100 mL three-necked flask, 35a (1.4 g, 4.9 mmol) dissolved in DCM (15 mL) was added, followed by triethylamine (0.6 g, 5.9 mmol), and then benzyl chloroformate (0.8 g, 4.9 mmol). The mixture was magnetically stirred at room temperature for 2 h. After the reaction was complete, the mixture was evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 3:1, v / v) to give an oily 35b (1.5 g, 73% yield). ESI-MS: m / z = 419 [M+1] + .

[0390] Step 3. Synthesis of benzyl (1-(3-(hydroxymethyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 35c)

[0391] The synthesis method for 35c is the same as that for 33a, except that 16b is replaced with 35b, yielding oily 35c (1.2 g, 87% yield). ESI-MS: m / z = 391 [M+1] + .

[0392] Step 4. Synthesis of benzyl (1-(3-formylnaphthalene-2-yl)piperidin-4-yl)carbamate (intermediate 35d)

[0393] The synthesis method for 35d is the same as that for 33b, except that 33a is replaced with 35a, yielding oily 35d (0.9 g, 75% yield). ESI-MS: m / z = 389 [M+1] + .

[0394] Step 5. Synthesis of benzyl(E)-(1-(3-(2-methoxyvinyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 35e)

[0395] In a 100 mL three-necked flask, 2.0 g (5.7 mmol) of (methoxymethyl)triphenylphosphine chloride dissolved in 10 mL of THF was added. The mixture was cooled to 0–5 °C, and then a 1.0 mol / L LiHMDS solution in THF was added dropwise. After the addition was complete, the reaction was maintained at this temperature for 0.5 h. Then, a 5 mL THF solution of 0.88 g (2.2 mmol) of 35 d was added dropwise, and the reaction was carried out at room temperature for 3 h. After the reaction was complete, saturated NH4Cl solution was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 4:1, v / v) to give an oily 35e (572 mg, yield 62%). ESI-MS: m / z = 417 [M+1] + .

[0396] Step 6. Synthesis of benzyl (1-(3-(2-oxoethyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 35f)

[0397] 35e (200 mg, 0.48 mmol) was dissolved in a mixture of acetone (4 ml) and H₂O (1 ml) in a 100 ml three-necked flask. The mixture was cooled to 0–5 °C, and then 2.8 ml of 40% HBr aqueous solution was added dropwise. After the addition was complete, the mixture was kept at this temperature for 3 h. After the reaction was complete, NaHCO₃ solution was added, and the mixture was extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (PE:EA = 4:1, v / v) to give an oily 35f (90 mg, yield 46%). ESI-MS: m / z = 403 [M+1] + .

[0398] Step 7. Synthesis of 2-(3-(4-((benzyloxy)carbonyl)amino)piperidin-1-yl)naphth-2-yl)acetic acid (intermediate 35 g)

[0399] In a 100 mL three-necked flask, 35f (150 mg, 0.36 mmol) was dissolved in tert-butanol (8 mL) and H₂O (2 mL). The mixture was cooled to 0–5 °C, and then 2,3-dimethyl-2-butene (610 mg, 7.20 mmol), potassium dihydrogen phosphate (102 mg, 0.72 mmol), and sodium chlorite (82 mg, 0.90 mmol) were added. After the addition was complete, the mixture was kept at this temperature and magnetically stirred for 3 h. After the reaction was complete, the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel column chromatography (DCM:MeOH = 25:1, v / v) to give 35 g (90 mg, 60% yield) of an oil. ESI-MS: m / z = 419 [M+1] + .

[0400] Step 8. Synthesis of benzyl (1-(3-(2-((2-fluorophenyl)amino)-2-oxoethyl)naphth-2-yl)piperidin-4-yl)carbamate (intermediate 35h)

[0401] The synthesis method for 35h is the same as that for 1c, except that 1b is replaced with 35g and N-methylbenzylamine is replaced with o-fluoroaniline, yielding oily 35h (0.05g, yield 43%). ESI-MS: m / z = 512 [M+1] + .

[0402] Step 9. Synthesis of 2-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-N-(2-fluorophenyl)acetamide (compound 35)

[0403] Add 35h (100 mg, 0.20 mmol) dissolved in AcOH (3 ml) to a 100 ml three-necked flask, add 40% HBr aqueous solution (2.5 ml), heat to 45 °C, and stir magnetically for 6 h. After the reaction is complete, cool to room temperature, add saturated sodium bicarbonate solution, extract with ethyl acetate, combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, evaporate to dryness using a rotary evaporator, and purify the residue by thin-layer chromatography (DCM:MeOH = 20:1, v / v) to give an oily compound 35 (20 mg, yield 27%).

[0404] 1H NMR (400MHz, CDCl3): δ8.31(d,J=6.3Hz,1H),8.15(t,J=8.1Hz,1H),7.82(s,1H),7.78-7.71(m,2H),7.52(s,1H),7.47-7.38(m,2H),7.08-7. 01(m,1H),6.97-6.91(m,2H),3.89(s,2H),3.31-3.13(m,3H),2.90-2. 79(m,2H),2.27-2.11(m,2H),2.02-1.95(m,4H).ESI-MS:m / z=378[M+1] + .

[0405] Preparation Example 36. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2,4-dimethylthiazolyl-5-yl)methyl)-N-methyl-2-naphthamide (compound 36)

[0406] Step 1. Synthesis of 1-(2,4-dimethylthiazol-5-yl)-N-methylmethylamine (intermediate 36a)

[0407] The synthesis method for 36a is the same as that for 33c, except that 33b is replaced with 2,4-dimethylthiazol-5-carboxaldehyde, yielding oily 36a (1.0 g, 46% yield). ESI-MS: m / z = 157 [M+1] + .

[0408] Step 2. Synthesis of tert-butyl carbamate (intermediate 36b)

[0409] The synthesis method for 36b is the same as that for 17c, except that 17b is replaced by 16d and N-methyl-2-fluorobenzylamine is replaced by 36a, yielding oily 36b (2.1 g, yield 63%). ESI-MS: m / z = 509 [M+1] + .

[0410] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2,4-dimethylthiazolyl-5-yl)methyl)-N-methyl-2-naphthamide (compound 36)

[0411] The synthesis method of compound 36 is the same as that of compound 17, except that 17c is replaced with 36b, to obtain solid compound 36 (1.4g, yield 83%).

[0412] 1H NMR (500MHz, CDCl3): δ7.91-7.84 (m, 2H), 7.73 (dt, J = 7.5, 2.0Hz, 1H), 7.51 -7.45(m,2H),7.42(d,J=2.0Hz,1H),7.36-7.30(m,1H),4.94(d,J=14.9Hz,1 H),4.72(d,J=14.9Hz,1H),3.56-3.41(m,2H),3.36-3.24(m,4H),3.12-3.00 (m,4H),2.61(s,3H),2.35(s,3H),1.88-1.62(m,4H).ESI-MS:m / z=409[M+1] + .

[0413] Preparation Example 37. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2,5-dimethylthiazolyl-4-yl)methyl)-N-methyl-2-naphthamide (compound 37)

[0414] Step 1. Synthesis of 1-(2,5-dimethylthiazol-4-yl)-N-methylmethylamine (intermediate 37a)

[0415] The synthesis method for 37a is the same as that for 33c, except that 33b is replaced with 2,5-dimethylthiazol-4-carboxaldehyde, yielding oily 37a (1.2 g, 53% yield). ESI-MS: m / z = 157 [M+1] + .

[0416] Step 2. Synthesis of tert-butyl carbamate (intermediate 37b)

[0417] The synthesis method for 37b is the same as that for 17c, except that 17b is replaced by 16d and N-methyl-2-fluorobenzylamine is replaced by 37a, yielding oily 37b (1.8 g, 48% yield). ESI-MS: m / z = 509 [M+1] + .

[0418] Step 3. Synthesis of 3-(4-aminopiperidin-1-yl)-N-((2,5-dimethylthiazolyl-4-yl)methyl)-N-methyl-2-naphthamide (compound 37)

[0419] The synthesis method of compound 37 is the same as that of compound 17, except that 17c is replaced with 37b, to obtain solid compound 37 (1.1g, yield 76%).

[0420] 1 H NMR (500MHz, CDCl3): δ7.89-7.86(m,2H),7.77-7.74(m,1H),7.51-7.45(m,2H),7.35-7.30(m,1H),4.90(d,J=14.3Hz,1H),4.72(d,J=14.3Hz, 1H),3.52-3.37(m,2H),3.36-3.27(m,2H),3.23(s,2H),3.18-3.02(m,4 H),2.61(s,3H),2.30(s,3H),1.78-1.59(m,4H).ESI-MS:m / z=409[M+1] + .

[0421] Preparation Example 38. Synthesis of 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidine-4-carboxamide (compound 38)

[0422] Step 1. Synthesis of tert-butyl 1-(3-(ethoxycarbonyl)naphth-2-yl)piperidine-4-carboxylic acid (intermediate 38a)

[0423] The synthesis method for 38a is the same as that for 16c, except that the starting material 4-tert-butoxycarbonylaminopiperidine is replaced with tert-butyl 4-piperidinecarboxylate, yielding oily 38a (0.2 g, 90% yield). ESI-MS: m / z = 384 [M+1] + .

[0424] Step 2. Synthesis of 3-(4-(tert-Butoxycarbonyl)piperidin-1-yl)-2-naphthoic acid (intermediate 38b)

[0425] The synthesis method for 38b is the same as that for 1b, except that raw material 1a is replaced with 38a, yielding solid 38b (0.2 g, yield 86%). ESI-MS: m / z = 356 [M+1] + .

[0426] Step 3. Synthesis of 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidine-4-carboxylic acid tert-butyl ester (intermediate 38c)

[0427] The synthesis method for 38c is the same as that for 16e, except that the starting material 16d is replaced with 38b, yielding oily 38c (0.2 g, 85% yield). ESI-MS: m / z = 477 [M+1] + .

[0428] Step 4. Synthesis of 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidine-4-carboxylic acid (intermediate 38d)

[0429] 0.2 g (0.4 mmol) of 38d was dissolved in 10 ml DCM in a 100 ml three-necked flask, followed by 1 ml TFA. The mixture was magnetically stirred at room temperature for 3 h. After the reaction was complete, the solution was diluted with water and extracted with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator to obtain solid 38d (135 mg, 80% yield). ESI-MS: m / z = 421 [M+1] + .

[0430] Step 5. Synthesis of 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidine-4-carboxamide (compound 38)

[0431] In a 50 mL three-necked flask, 38d (135 mg, 0.3 mmol) dissolved in DMF (10 mL) was added, along with HATU (260 mg, 0.5 mmol) and TEA (91 mg, 0.9 mmol). The mixture was magnetically stirred at room temperature for 1 h, followed by the addition of NH4Cl (50 mg, 0.6 mmol) and magnetic stirring for 3 h. After the reaction was complete, the mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and evaporated to dryness using a rotary evaporator. The residue was purified by silica gel chromatography (DCM:MeOH = 10:1, v / v) to give an oily compound 38 (40 mg, 32% yield).

[0432] 1 H NMR (400MHz, CDCl3): δ7.82-7.70(m,3H),7.66-7.60(m,1H),7.50-7.35(m,4H) ,7.29-6.95(m,2H),5.12-4.73(m,1H),4.40(dt,J=54.9,16.4Hz,1H),3.75-3.5 5(m,1H),3.45-3.21(m,1H),3.15-3.00(m,2H),2.90-2.74(m,3H),2.65-2.40(m ,1H),2.20-1.85(m,2H),1.74(s,1H),1.68-1.35(m,1H).ESI-MS:m / z=420[M+1] + .

[0433] Preparation Example 39. Synthesis of N-(2-fluorobenzyl)-N-methyl-3-piperidine-4-amino-2-naphthamide (compound 39)

[0434] Step 1. Synthesis of tert-butyl 4-(3-(ethoxycarbonyl)naphthalene-2-amino)piperidine-1-carboxylic acid (intermediate 39a)

[0435] The synthesis method for 39a is the same as that for 16c, except that the starting material 4-tert-butoxycarbonylaminopiperidine is replaced with 1-Boc-4-aminopiperidine, yielding oily 39a (2.4 g, 85% yield). ESI-MS: m / z = 399 [M+1] + .

[0436] Step 2. Synthesis of 3-((1-(tert-Butoxycarbonyl)piperidin-4-yl)amino)-2-naphthoic acid (intermediate 39b)

[0437] The synthesis method of 39b is the same as that of 1b, except that the starting material 1a is replaced with 39a, yielding solid 39b (1.9 g, yield 83%). ESI-MS: m / z = 371 [M+1] + .

[0438] Step 3. Synthesis of 4-((3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)amino)piperidine-1-carboxylic acid tert-butyl ester (intermediate 39c)

[0439] The synthesis method for 39c is the same as that for 16e, except that the starting material 16d is replaced with 39b, yielding oily 39c (2.4 g, 95% yield). ESI-MS: m / z = 492 [M+1] + .

[0440] Step 4. Synthesis of N-(2-fluorobenzyl)-N-methyl-3-piperidine-4-amino-2-naphthamide (compound 39)

[0441] The synthesis method of compound 39 is the same as that of compound 1, except that 1c is replaced with 39c, to obtain solid compound 39 (1.1g, yield 60%).

[0442] 1H NMR (400MHz, CDCl3): δ7.89-7.75(m,3H),7.70-7.63(m,1H),7.58-7.35(m,4H) ,7.29-6.95(m,2H),5.12-4.82(m,1H),4.46(dt,J=54.9,16.4Hz,1H),3.85-3.6 2(m,1H),3.48-3.23(m,1H),3.15-3.00(m,2H),2.92-2.70(m,3H),2.65–2.57(m ,1H),2.25-1.95(m,2H),1.82(s,1H),1.30-1.20(m,1H).ESI-MS:m / z=392[M+1] + .

[0443] Preparation Example 40. Synthesis of 3-(4-aminopiperidin-1-yl)-N-(2-cyanobenzyl)-N-methyl-2-naphthamide (Compound 40)

[0444] Step 1. Synthesis of tert-butyl carbamate (intermediate 40a)

[0445] The synthesis method for 40a is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 2-((methylamino)methyl)benzonitrile, yielding oily 40a (2.3 g, 85% yield). ESI-MS: m / z = 499 [M+1] + .

[0446] Step 2. Synthesis of 3-(4-aminopiperidin-1-yl)-N-(2-cyanobenzyl)-N-methyl-2-naphthamide (compound 40)

[0447] The synthesis method of compound 40 is the same as that of compound 1, except that 1c is replaced with 40a, to obtain solid compound 40 (1.3g, yield 70%).

[0448] 1H NMR (400MHz, CDCl3): δ7.80-7.70(m,3H),7.65-7.58(m,1H),7.50-7.30(m,4H) ,7.25-6.90(m,3H),5.00-4.73(m,1H),4.35(dt,J=54.9,16.4Hz,1H),3.75-3.5 5(m,1H),3.40-3.10(m,1H),3.15-3.00(m,2H),2.85-2.67(m,3H),2.50-2.48(m ,1H),2.15-1.78(m,2H),1.72(s,1H),1.33-1.20(m,1H).ESI-MS:m / z=399[M+1] + .

[0449] Preparation Example 41. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(isoindoline-2-yl)methyl ketone (compound 41)

[0450] Step 1. Synthesis of (1-(3-(isoindoline-2-carbonyl)naphth-2-yl)piperidin-4-yl)tert-butyl carbamate (intermediate 41a)

[0451] The synthesis method for 41a is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with isoindolin, yielding oily 41a (2.3 g, yield 89%). ESI-MS: m / z = 472 [M+1] + .

[0452] Step 2. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(isoindoline-2-yl)methyl ketone (compound 41)

[0453] The synthesis method of compound 41 is the same as that of compound 1, except that 1c is replaced with 41a, to obtain solid compound 41 (1.3g, yield 75%).

[0454] 1H NMR (400MHz, CDCl3): δ8.18–8.13(m,1H),7.88-7.82(m,1H),7.74(d,J=6.8Hz,1H),7 .57-7.48(m,2H),7.32-7.25(m,2H),7.24-7.19(m,1H),7.13-7.09(m,1H),7.01-6.8 7(m,1H),5.07(d,J=7.3Hz,2H),4.88-4.81(m,2H),3.56-3.46(m,2H),3.42-3.28(m, 1H),2.96-2.78(m,2H),2.35-2.24(m,2H),2.20-2.06(m,2H).ESI-MS:m / z=372[M+1] + .

[0455] Preparation Example 42. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(4-fluoroisoindoline-2-yl)methyl ketone (compound 42)

[0456] Step 1. Synthesis of tert-butyl carbamate (intermediate 42a)

[0457] The synthesis method for 42a is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 4-fluoroisoindoline, yielding oily 42a (2.3 g, yield 87%). ESI-MS: m / z = 490 [M+1] + .

[0458] Step 2. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(4-fluoroisoindololin-2-yl)methyl ketone (compound 42)

[0459] The synthesis method of compound 42 is the same as that of compound 1, except that 1c is replaced with 42a, to obtain solid compound 42 (1.5g, yield 80%).

[0460] 1H NMR (400MHz, CDCl3): δ8.47(m,2H),7.75(dd,J=19.3,11.0Hz,2H),7.48(ddd,J=23.7,13.2,8.2Hz,2H),7.36-7.27(m,1H),7.22-7.01(m,2H ),4.96(s,1H),4.54(m,1H),3.69-3.21(m,1H),2.90(m,1H),2.20(d,J=28.1Hz,1H),1.91(m,5H),1.35-1.17(m,2H).ESI-MS:m / z=390[M+1] + .

[0461] Preparation Example 43. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 43)

[0462] Step 1. Synthesis of tert-butyl carbamate (intermediate 43a) of (1-(3-(1,2,3,4-tetrahydroisoquinoline-2-carbonyl)naphth-2-yl)piperidin-4-yl)carbamate

[0463] The synthesis method for 43a is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 1,2,3,4-tetrahydroisoquinoline, yielding oily 43a (2.2 g, 83% yield). ESI-MS: m / z = 486 [M+1] + .

[0464] Step 2. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 43)

[0465] The synthesis method of compound 43 is the same as that of compound 1, except that 1c is replaced with 43a, to obtain solid compound 43 (1.4g, yield 83%).

[0466] 1 H NMR (400MHz, CDCl3): δ8.56–8.27(m,2H),7.81-7.59(m,2H),7.53-7.34(m,3H),7.18-6 .95(m,3H),5.08-3.86(m,2H),3.77-2.53(m,7H),2.42-1.93(m,3H),1.69-1.14(m,3H). ESI-MS:m / z=386[M+1] + .

[0467] Preparation Example 44. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(8-fluoro-3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 44)

[0468] Step 1. Synthesis of tert-butyl carbamate (intermediate 44a) of 1-(3-(8-fluoro-1,2,3,4-tetrahydroisoquinoline-2-carbonyl)naphth-2-yl)piperidin-4-yl)carbamate

[0469] The synthesis method for 44a is the same as that for 16e, except that the starting material N-methyl-2-fluorobenzylamine is replaced with 8-fluoro-1,2,3,4-tetrahydroisoquinoline, yielding oily 44a (1.9 g, 87% yield). ESI-MS: m / z = 404 [M+1] + .

[0470] Step 2. Synthesis of (3-(4-aminopiperidin-1-yl)naphth-2-yl)(8-fluoro-3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 43)

[0471] The synthesis method of compound 44 is the same as that of compound 1, except that 1c is replaced with 44a, to obtain solid compound 44 (1.7g, yield 89%).

[0472] 1 H NMR (400MHz, CDCl3): δ7.82-7.72(m,3H),7.51-7.44(m,1H),7.42-7.36(m,1H),7.35-7.32(m,1H),7.20-7.11(m,1H),7.02-6.80(m,2H), 5.31-4.55(m,1H),4.46-4.09(m,1H),3.80-3.65(m,1H),3.62-2.42(m,7H),1.94-1.68(m,2H),1.61-1.09(m,3H).ESI-MS: m / z=404[M+1] + .

[0473] Biological assessment

[0474] Test Example 1. Inhibitory Activity Assay of TRPC6 Channel (Electrophysiological Assay)

[0475] Experimental methods

[0476] Human embryonic kidney cells HEK-293 (obtained from the Cell Bank of the Chinese Academy of Sciences) were cultured in Dulbecco's Modified Eagle's medium (DMEM, obtained from Gibco) supplemented with 10% fetal bovine serum (FBS, obtained from PAM-Biotech GmbH) at 37°C in an incubator with 95% air and 5% carbon dioxide. Human TRPC6 (hTRPC6) plasmid (obtained from BGI Genomics) was transfected into HEK-293 cells using a calcium phosphate transfection solution containing two types of solutions: Solution A was pure water containing 250 mM CaCl2, and Solution B contained 1.5 mM Na2HPO4, 140 mM NaCl, and 50 mM HEPES (adjusted to pH 6.96). All electrophysiological data were recorded at room temperature using an Axon 200B (obtained from Molecular Devices) 24–48 hours post-transfection, maintaining a potential of -60 mV. When filling an intracellular solution containing 110 mM CsCl, 10 mM HEPES, 10 mM BAPTA, 1 mM MgCl2 and 4.77 mM CaCl2 (adjusted to pH 7.2 with HCl) (free Ca 2+ The concentration is 400 nM, and the resistance range of the patch electrode is 3-5 MΩ. When testing the hTRPC6 channel, the standard electrode solution is a solution containing 110 mM CsCl, 10 mM HEPES, 10 mM BAPTA, 1 mM MgCl2 and 4.77 mM CaCl2 (adjusted to pH 7.2 with HCl) (free Ca). 2+ The concentration was 400 nM; the standard extracellular solution was a solution containing 150 mM NaCl, 5 mM KCl, 10 mM glucose, 2 mM CaCl2, 1 mM MgCl2, and 10 mM HEPES (pH adjusted to 7.4 using Tris-base). Data were sampled at 10 kHz and filtered at 2 kHz. All test compounds were pre-administered for 30 seconds and then co-administered in the presence of agonist M085 (obtained from Hunan Yaowei Biomedical Co., Ltd.), and the current magnitude was recorded. The current when agonist M085 was administered alone served as a control.

[0477] The current when agonist M085 is used alone is denoted as I. max The current when the test compound and agonist M085 are co-administered is recorded as I. The ratio of I / I is used to express the current. max A concentration-effect curve was plotted with the test compound concentration on the x-axis and the test compound concentration on the y-axis. The IC50 of the test compound for the hTRPC6 receptor was calculated using the Hill formula. 50 value.

[0478] The inhibitory activity data (IC50) of the test compounds against hTRPC6 obtained by the above method were used. 50 The values ​​are shown in Table 1 below.

[0479] Table 1. Inhibitory activity data of the tested compounds against hTRPC6

[0480] As can be seen from the results in Table 1, the compounds of the present invention, particularly compounds 1, 4, 8, 16, 23 and 25, have good inhibitory activity against TRPC6 and therefore may be used for the prevention or treatment of TRPC6-mediated diseases.

[0481] Test Example 2. Inhibitory Activity Tests on Multiple Channels (Electrophysiological Assay)

[0482] The inhibitory activity of compound 16 against TRPC6, TRPC3, TRPC5, TRPV3, TRPM8, and TRPA1 channels was investigated using the method described in Test Example 1. Specifically, plasmids mTRPC6, hTRPC3, hTRPC5, hTRPV3, mTRPM8, and mTRPA1 (all obtained from BGI Genomics) were transfected into HEK-293 cells via transient transfection, and electrophysiological assays were performed to test the inhibitory activity.

[0483] For the mTRPC6 channel inhibitory activity assay, the standard electrode solution and standard extracellular solution are the same as those used in the hTRPC6 channel inhibitory activity assay. For the TRPC3 and TRPC7 channel inhibitory activity assays, the standard extracellular solution is adjusted to contain 150 mM NaCl, 5 mM KCl, 10 mM glucose, 2 mM CaCl2, 1 mM MgCl2, and 10 mM HEPES (adjusted to pH 7.4 with Tris-base). For the TRPC5 channel inhibitory activity assay, the standard electrode solution is adjusted to contain 130 mM CsCl, 1 nM MgCl2, 5.7 nM CaCl2, 10 mM EGTA, and 10 mM HEPES; the standard extracellular solution is adjusted to contain 140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 2 mM CaCl2, 10 mM HEPES, and 10 mM glucose. For the TRPV3, TRPA1, and TRPM8 channel activity inhibition assays, the standard electrode solution was adjusted to contain 140 mM CsCl, 10 mM HEPES, and 5 mM EGTA (pH adjusted to 7.4 with CsOH); the standard extracellular solution was adjusted to contain 140 mM NaCl, 5 mM KCl, 1 mM MgCl2, 10 mM glucose, and 10 mM HEPES (pH adjusted to 7.2 with NaOH).

[0484] The inhibitory activity data (IC50) of compound 16 against multiple channels obtained by the above method were used. 50 The values ​​are shown in Table 2 below.

[0485] Table 2. Inhibitory activity data of the tested compounds on multiple channels

[0486] As shown in Table 2, the IC50 values ​​of the compounds of this invention for inhibiting TRPC6 activity are [not specified in the original text]. 50 The value is much lower than the IC50 value for inhibiting the activity of other ion channels such as TRPC3, TRPC5, TRPV3, TRPM8, and TRPA1. 50 The values ​​indicate that the inhibitory activity of the compounds of this invention against TRPC6 can be tens or even thousands of times stronger than their inhibitory activity against TRPC3, TRPC5, TRPV3, TRPM8, and TRPA1. These results demonstrate that the compounds of this invention exhibit high selectivity for TRPC6 compared to other ion channels.

[0487] Therefore, the compounds of the present invention can specifically inhibit the TRPC6 channel, which is expected to provide a new strategy for the prevention and treatment of TRPC6-related diseases. Furthermore, the application of the compounds of the present invention to prevent or treat TRPC6-related diseases can reduce interference with the body's normal physiological functions, thereby reducing the side effects of traditional drug treatments and improving the safety of treatment.

[0488] Test Example 3. Functional Analysis of TRPC6 Inhibition in MPC5 Cells of Mouse Podocytes

[0489] Experimental methods

[0490] 1. Culture and induction of differentiation of mouse glomerular podocytes MPC5 (purchased from ATCC)

[0491] Mouse podocyte MPC5 culture conditions: cultured in RPMI 1640 basal medium (obtained from GIBCO) supplemented with 10 U / mL mIFN-γ (obtained from PeproTech), 10% fetal bovine serum (obtained from GIBCO), and 1% penicillin-streptomycin (obtained from GIBCO) in a saturated humidity incubator at 33°C and 5% CO2.

[0492] Mouse podocyte MPC5 differentiation culture conditions: Without the addition of cytokine mIFN-γ, RPMI 1640 basal culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin was used for cytokine-free induction differentiation culture in a saturated humidity incubator at 37°C and 5% CO2. After 10-14 days of cytokine-free induction differentiation, the cells differentiated into functional podocytes (approximately 90% or more).

[0493] Podocyte morphology was observed (200x) using a microscope (DMi8) obtained from a Leica microscopy system. Figures 1 and 2 show undifferentiated and differentiated MPC5 cells, respectively. As shown in Figure 2, compared to undifferentiated MPC5 cells (Figure 1), differentiated MPC5 cells exhibit the morphology of mouse podocytes with prominent foot processes. Differentiated MPC5 podocytes can normally express TRPC6 channels and possess Ca2+. 2+ Regulatory function.

[0494] 2. MPC5 cell proliferation activity assay

[0495] Differentiated mouse podocyte MPC5 cell suspensions were prepared and counted. Approximately 100 μL of the cell suspension was seeded into each well of a 96-well plate, with three replicates per well. The plates were incubated overnight at 37°C, 5% CO2. Mouse podocyte MPC5 cells were treated with the agonist M085, or a combination of the agonist and the compound of this invention, for 48 h. 10 μL of CCK-8 solution (obtained from Jiangsu Kaiji Biotechnology) was added to each well, and the plates were incubated for 1 h. The absorbance (OD) at 450 nm was measured using a Varioskan™ LUX microplate reader (obtained from Thermo Fisher Scientific Inc.). Relative absorbance values ​​were calculated to represent relative cell viability, using a control group (Ctrl) (0.1% DMSO or PBS) as a reference.

[0496] 3. Intracellular Ca 2+ Level (Fluo-4 fluorescence intensity) detection

[0497] Differentiated mouse podocytes (MPC5) were seeded into 24-well plates and cultured overnight at 37°C until well adhered. Cells were treated with 10 μM of the agonist M085 for 0 (untreated), 1 h, 12 h, and 24 h. After each treatment, 75 μL of trypsin (from GIBCO) was added to digest the cells for 2 minutes, and the treatment was terminated with 500 μL of complete culture medium (RPMI 1640 basal medium containing 10% fetal bovine serum and 1% penicillin-streptomycin). The culture was collected in 1.5 mL EP tubes and centrifuged at 1000 rpm for 5 minutes. The supernatant was discarded, and the cells were washed once with 1 mL of PBS. After centrifugation, 100 μL of 4 μM Fluo-4 AM working solution (from Dojindo) was added, and the cells were incubated at 37°C in the dark for 30 minutes. Incubation was terminated with 500 μL of PBS, and the cells were centrifuged at 1000 rpm for 5 minutes, and the supernatant was discarded. Add 500 μL of PBS to the cell pellet, gently mix, and incubate at 37°C in the dark for 30 minutes to ensure that Fluo-4AM is completely converted into Fluo-4 in the cells. Finally, collect the cells using a flow cytometer (LSR-Fortessa) obtained from BD.

[0498] Experimental results

[0499] 1. The damaging activity of agonist M085 against MPC5

[0500] In the above-mentioned MPC5 cell proliferation activity assay, mouse podocyte MPC5 cells were treated with M085 (obtained from Hunan Yaowei Biomedical Co., Ltd.) at concentrations of 3 μM, 10 μM, and 30 μM for 48 h. The relative cell viability after treatment with different concentrations of the agonist M085 for 48 hours is shown in Figure 3. One-way ANOVA (Bonferroni test) was used in this experiment. Data are expressed as mean ± standard deviation (Mean ± SD), * indicates P < 0.05, and *** indicates P < 0.001.

[0501] As shown in Figure 3, the TRPC6 agonist M085 inhibited the viability of MPC5 cells in a concentration-dependent manner, and significantly induced damage to mouse podocyte MPC5 cells by activating TRPC6 at concentrations of 10 μM and 30 μM.

[0502] In the above-mentioned cells, Ca 2+In the level (Fluo-4 fluorescence intensity) assay, cells were treated with 10 μM M085 (obtained from Hunan Yaowei Biomedical Co., Ltd.) for 0 (untreated), 1 h, 12 h, and 24 h. The Fluo-4 fluorescence intensities obtained after treatment with the agonist M085 for different times are shown in Figure 4. One-way ANOVA (Bonferroni test) was used in this experiment. Data are expressed as mean ± standard deviation (Mean ± SD), * indicates P < 0.05, ** indicates P < 0.01, and ns indicates no difference.

[0503] As shown in Figure 4, the TRPC6 agonist M085 (10 μM) can continuously increase intracellular calcium levels starting from 1 hour. 2+ At the level, the fluorescence intensity of Fluo-4 gradually increased from 29.9±4.0% (0 hours) to 31.6±1.9% (1 hour), 37.5±4.5% (12 hours) and 40.4±1.1% (24 hours).

[0504] The above results confirm that the TRPC6 agonist M085 increases intracellular calcium ions (Ca) by directly activating the TRPC6 channel. 2+ The level of ) can induce podocyte damage.

[0505] 2. The protective effect of the compounds of the present invention against MO85-induced podocyte injury.

[0506] In the above-described MPC5 cell proliferation assay, cells were treated with M085 alone (10 μM) or simultaneously with different concentrations of compound 25 (2.22 μM, 0.74 μM, 0.24 μM, 0.082 μM, 0.027 μM, and 0.009 μM) for 48 hours. The relative cell viability after 48 hours of treatment with M085 alone (10 μM) and simultaneously with different concentrations of compound 25 is shown in Figure 5. One-way ANOVA (Bonferroni test) was used in this study. Data are expressed as mean ± standard deviation (Mean ± SD), * indicates P < 0.05.

[0507] As shown in Figure 5, compound 25 at 0.74 μM and 2.22 μM could partially restore the damage to podocytes caused by 10 μM M085, and cell viability increased from 60.2 ± 2.60% (M085 group) to 75.5 ± 10.0% (0.74 μM compound 25 + M085 group) and 79.6 ± 10.6% (2.22 μM compound 25 + M085 group), respectively.

[0508] The above results verify that the compound of the present invention can effectively inhibit the activity of TRPC6, thereby having a restorative effect on podocyte damage induced by TRPC6 overactivation.

[0509] Test Example 4. Pharmacokinetic Analysis

[0510] Adult male SD rats (weighing 200–220 g, obtained from Shanghai Silex Laboratory Animal Co., Ltd.) were housed for 3 days to acclimatize to their environment. After fasting but allowing free access to water for approximately 12 hours, animal experiments were conducted, followed by refeeding 4 hours after drug administration. Compound 16 was administered to rats via intravenous injection (IV, 1.5 mg / kg) and oral gavage (IG, 15 mg / kg). Blood samples were collected from the tail vein of the rats at different time points after administration, and plasma drug concentrations were measured using LC-MS / MS. The drug-time curves of compound 16 in SD rat plasma after intravenous injection and oral gavage administration are shown in Figures 6 and 7, respectively.

[0511] Following intravenous (IV) administration of compound 16 at a dose of 1.5 mg / kg, the plasma clearance (CL) of compound 16 in rats was 18.2 mL / min / kg (1.09 L / h / kg), which is 26.7% of the hepatic blood flow (68 mL / min / kg in rats). These results indicate that compound 16 has a low plasma clearance in rats.

[0512] Following a single oral (IG) administration of compound 16 at a dose of 15 mg / kg, the T levels of compound 16 in rat plasma were [not specified]. max It is 4.67hr, C max The concentration was 731.05 ng / mL, and the bioavailability was 99%. These results indicate that compound 16 exhibits favorable pharmacokinetic characteristics in rats, including low clearance, high metabolic stability, and excellent oral bioavailability, and thus possesses high clinical translational value.

[0513] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. It should be noted that those skilled in the art can make various modifications, improvements, refinements, and equivalent substitutions without departing from the principles and rules of the present invention, and all such modifications, improvements, refinements, and equivalent substitutions should be considered within the scope of protection of the present invention.

[0514] Industrial application

[0515] This invention provides a TRPC6 inhibitor compound or a pharmaceutically acceptable salt thereof, and also provides uses and methods for the prevention or treatment of TRPC6-related diseases using the aforementioned compound or pharmaceutically acceptable salt or prodrug. Therefore, the aforementioned TRPC6 inhibitor can be formulated into corresponding pharmaceuticals suitable for industrial application.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof: in: The A ring is a 4-10 member nitrogen-containing heterocycle, spiroheterocycle, or bridged heterocycle, which is optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, amino, halogen, and -C(O)NH2, which are optionally substituted with amino groups. The B ring is a 6-10 membered monocyclic or bicyclic aromatic ring or a 6-10 membered monocyclic or bicyclic heteroaromatic ring containing a nitrogen atom, which is optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, C1-C6 alkoxy, halogen and cyano, and 5-membered heteroaryl containing 1-3 heteroatoms selected from N, S and O. n is 0 or 1; R is -C(O)-NR1R2 or -NR3-C(O)R4, wherein R1, R2, R3 and R4 are each independently selected from C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 aryl C1-C3 alkylene, and 5-8 heteroaryl C1-C3 alkylene, wherein the 5-8 heteroaryl contains 1-3 heteroatoms selected from N, S and O, wherein the 6-10 aryl or 5-8 heteroaryl is optionally substituted by one or more substituents selected from C1-C3 alkyl, halogen and cyano, or may form a 5-8 nitrogen-containing heterocyclic group fused with the benzene ring together with the attached nitrogen atom, wherein the benzene ring is optionally substituted with a halogen.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R is -C(O)-NR. 1a R 2a , where R 1a and R 2a Each is independently selected from C1-C6 alkyl, C3-C6 cycloalkyl, 6-10 aryl C1-C3 alkylene, and 5-8 heteroaryl C1-C3 alkylene, wherein the 5-8 heteroaryl contains 1-3 heteroatoms selected from N, S, and O, and the 6-10 aryl or 5-8 heteroaryl is optionally substituted by one or more substituents selected from C1-C3 alkyl, halogen, and cyano; or R 1a and R 2a Together with the attached nitrogen atom, it forms a 5-8 membered nitrogen-containing heterocyclic group fused with the benzene ring, wherein the benzene ring is optionally substituted with a halogen.

3. The compound of claim 2 or a pharmaceutically acceptable salt thereof, wherein R 1a It is a C1-C6 alkyl or C3-C6 cycloalkyl; R 2a It is a benzyl or 5-6-membered heteroarylmethylene group, wherein the 5-6-membered heteroaryl group contains 1-2 heteroatoms selected from N and S, wherein the benzyl or 5-6-membered heteroarylmethylene group is optionally substituted for one or more substituents selected from C1-C3 alkyl, halogen and cyano groups on the phenyl or 5-membered heteroaryl group; or R 1a and R 2a Together with the attached nitrogen atom, it forms a 5- or 6-membered nitrogen-containing heterocyclic group fused with the benzene ring, wherein the benzene ring is optionally substituted with F.

4. The compound according to claim 2 or 3, or a pharmaceutically acceptable salt thereof, wherein R 1a It is a C1-C3 alkyl or C3-C6 cycloalkyl, preferably methyl, isopropyl, or cyclopropyl, more preferably methyl; R 2a It is benzyl, pyridylmethylene or thiazolylmethylene, more preferably benzyl.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is represented by formula (II): The A ring is a 4-10 member nitrogen-containing heterocycle, spiroheterocycle, or bridged heterocycle, which is optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, amino, halogen, and -C(O)NH2, which are optionally substituted with amino groups. The B ring is a 6-10 membered monocyclic or bicyclic aromatic ring or a 6-10 membered monocyclic or bicyclic heteroaromatic ring containing a nitrogen atom, which is optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, C1-C6 alkoxy, halogen and cyano, and 5-membered heteroaryl containing 1-3 heteroatoms selected from N, S and O. R 1b It is a C1-C6 alkyl or C3-C6 cycloalkyl; R 2b It is a benzyl or 5-6-membered heteroarylmethylene group, wherein the 5-6-membered heteroaryl group contains 1-2 heteroatoms selected from N and S, wherein the benzyl or 5-6-membered heteroarylmethylene group is optionally substituted for one or more substituents selected from C1-C3 alkyl, halogen and cyano groups on the phenyl or 5-membered heteroaryl group; or R 1b and R 2b Together with the attached nitrogen atom, it forms a 5- or 6-membered nitrogen-containing heterocyclic group fused with the benzene ring, wherein the benzene ring is optionally substituted with F.

6. The compound of claim 5 or a pharmaceutically acceptable salt thereof, wherein R 1b It is a C1-C3 alkyl or C3-C6 cycloalkyl, preferably methyl, isopropyl, or cyclopropyl, more preferably methyl; R 2b It is benzyl, pyridylmethylene or thiazolylmethylene, more preferably benzyl.

7. The compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein ring A is a nitrogen-containing heterocycle, spiroheterocycle, or bridged heterocycle selected from the following structures:

8. A compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein the B ring is an aromatic or heteroaromatic ring selected from the following structures:

9. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from: 2-(4-aminopiperidin-1-yl)-N-benzyl-N-methylbenzamide (compound 1); 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-N-methylbenzamide (compound 2); 2-(4-aminoacrahepane-1-yl)-N-(4-fluorobenzyl)-N-methylbenzamide (compound 3); 2-(4-aminoacrahepane-1-yl)-3-fluoro-N-(2-fluorobenzyl)-N-methylbenzamide (compound 4); 2-(4-aminoacrahepane-1-yl)-N-(2-fluorobenzyl)-5-methoxy-N-methylbenzamide (compound 5); 2-(4-aminoacrahepane-1-yl)-3-cyano-N-(2-fluorobenzyl)-N-methylbenzamide (compound 6); 2-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,4-triazol-1-yl)benzamide (compound 7); 3-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 8); 2-(4-aminoachrogen-1-yl)-N-benzyl-3-fluoro-N-(2-fluorobenzyl)benzamide (compound 9); 3-(4-aminoacrahepane-1-yl)-3-fluoro-N-(2-fluorobenzyl)-N-isopropylbenzamide (compound 10); 2-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methyl-5-(1H-1,2,3-triazol-1-yl)benzamide (compound 11); 2-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methyl-4-(1H-1,2,3-triazol-1-yl)benzamide (compound 12); 2-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-3-carboxamide (compound 13); 7-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methylisoquinoline-6-carboxamide (compound 14); 7-(4-aminoacrheptan-1-yl)-N-(2-fluorobenzyl)-N-methylquinoline-6-carboxamide (compound 15); 3-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 16); 3-(3-aminopyrrolidone-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 17); 3-(3-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 18); 2-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-1-naphthamide (compound 19); 1-(4-aminopiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 20); 3-(3-(aminomethyl)azacyclobut-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthylcarboxamide (compound 21); 3-(6-amino-3-azabicyclo[3.1.1]hept-3-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 22); 3-(6-amino-2-azaspirocyclic[3.3]heptane-2-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 23); 3-(3-aminoazacyclobutane-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 24); 3-(3-aminomethyl)azacyclobutane-1-yl-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 25); 3-(4-amino-4-methylpiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 26); 3-(4-aminopiperidin-1-yl)-N-((2-fluoropyridin-3-yl)methyl)-N-methyl-2-naphthamide (compound 27); 3-(4-amino-3,3-difluoropiperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 28); 3-(4-(aminomethyl)piperidin-1-yl)-N-(2-fluorobenzyl)-N-methyl-2-naphthamide (compound 29); 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-4-yl)methyl)-N-methyl-2-naphthamide (compound 30); 3-(4-aminopiperidin-1-yl)-N-((3-fluoropyridin-2-yl)methyl)-N-methyl-2-naphthamide (compound 31); 3-(4-aminopiperidin-1-yl)-N-cyclopropyl-N-((3-fluoropyridin-2-yl)methyl)-2-naphthamide (compound 32); N-((3-(4-aminopiperidin-1-yl)naphth-2-yl)methyl)-2-fluoro-N-methylbenzamide (compound 33); N-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-2-(2-fluorophenyl)acetamide (compound 34); 2-(3-(4-aminopiperidin-1-yl)naphth-2-yl)-N-(2-fluorophenyl)acetamide (compound 35); 3-(4-aminopiperidin-1-yl)-N-((2,4-dimethylthiazolyl-5-yl)methyl)-N-methyl-2-naphthamide (compound 36); 3-(4-aminopiperidin-1-yl)-N-((2,5-dimethylthiazolyl-4-yl)methyl)-N-methyl-2-naphthamide (compound 37); 1-(3-((2-fluorobenzyl)(methyl)carbamoyl)naphth-2-yl)piperidine-4-carboxamide (compound 38); N-(2-Fluorobenzyl)-N-methyl-3-piperidine-4-amino-2-naphthamide (compound 39); 3-(4-aminopiperidin-1-yl)-N-(2-cyanobenzyl)-N-methyl-2-naphthamide (compound 40); (3-(4-aminopiperidin-1-yl)naphth-2-yl)(isoindoline-2-yl)methyl ketone (compound 41); (3-(4-aminopiperidin-1-yl)naphth-2-yl)(4-fluoroisoindololin-2-yl)methyl ketone (compound 42); (3-(4-aminopiperidin-1-yl)naphth-2-yl)(3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 43); and (3-(4-aminopiperidin-1-yl)naphth-2-yl)(8-fluoro-3,4-dihydroisoquinoline-2(1H)-yl)methyl ketone (compound 44).

10. A pharmaceutical composition comprising a compound according to any one of claims 1-9 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients; preferably, the pharmaceutical composition is in the form of oral administration.

11. Use of the compound of any one of claims 1-9 or a pharmaceutically acceptable salt thereof or the composition of claim 10 in the manufacture of a medicament for the prevention or treatment of TRPC6-related diseases; preferably, the medicament is in a dosage form suitable for oral administration.

12. The use according to claim 11, wherein the disease is selected from kidney and urinary system diseases, cardiovascular system diseases, neoplastic diseases and systemic functional disorders; preferably, the disease is selected from glomerular dysfunction diseases, metabolic nephropathy, acute / chronic kidney injury, tubulointerstitial lesions and secondary renal fibrosis, circulatory dysfunction diseases, myocardial remodeling-related diseases, vascular dysfunction diseases, rhythm abnormalities, thromboembolic diseases, solid tumors and hematologic malignancies, respiratory diseases and metabolic disorder-related diseases.

13. The disease according to claim 12, wherein the disease is selected from focal segmental glomerulosclerosis, membranous nephropathy, IgA nephropathy, diabetic nephropathy, hypertensive nephropathy, acute / chronic kidney injury, tubulointerstitial lesions, secondary renal fibrosis, primary and secondary hypertension, pulmonary hypertension, heart failure, myocardial hypertrophy, myocarditis, atherosclerosis, vascular calcification, arrhythmia, thromboembolic diseases, ischemic stroke, gastric adenocarcinoma, lung cancer, breast cancer, prostate cancer, hematologic malignancies, insulin resistance syndrome, and non-alcoholic steatohepatitis.

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