3-amido-beta-carboline compounds, methods for their preparation and uses

Abstract

The present application relates to a kind of 3-amido-β-carboline compound, its preparation method and purposes, specifically related to 3-amido-β-carboline compound represented by general formula P, its optical isomer, pharmaceutically acceptable salt and hydrate, its preparation method, pharmaceutical composition comprising it and its pharmaceutical purposes.The 3-amido-β-carboline compound of the present application, its optical isomer, pharmaceutically acceptable salt and hydrate can be inhibited by RANKL-RANK interaction, intervene OPG-RANKL-RANK signal system, regulate RANKL activity in osteoclast precursor cell and inhibit the formation of osteoclast, thereby reducing bone absorption.The compound of the present application, its optical isomer, pharmaceutically acceptable salt and hydrate are expected to play the role of preventing and treating bone metabolism disease.

Description

Technical Field

[0001] This invention relates to osteoclast inhibitors. Specifically, this invention relates to a class of 3-amido-β-carboline compounds, their optical isomers, pharmaceutically usable salts and hydrates, methods for their preparation, and their use in the preparation of medicaments for the prevention and / or treatment of diseases caused by abnormal osteoclast activity. Background Technology

[0002] Osteoclasts are the only bone resorption cells in the human body. Their bone resorption function plays a crucial role in maintaining bone metabolic homeostasis, helping to maintain the integrity of the skeletal system and mineral homeostasis, and is closely related to local or systemic bone metabolic diseases. Osteoclasts originate from mononuclear myeloid hematopoietic stem cells in the bone marrow and are specifically differentiated macrophages, differentiated by stimulation of macrophage clonal stimulating factor (M-CSF) and nuclear factor κB ligand-activated receptor (RANKL). In osteoclast precursor cells, M-CSF binds to its receptor c-Fos, providing the necessary signal for the proliferation of osteoclast precursor cells; RANKL binds to RANK, thereby initiating the differentiation of precursor osteoclasts into mature osteoclasts.

[0003] In normal bone metabolism, bone regeneration is a coordinated process involving osteoblasts and osteoclasts. Osteogenesis and resorption are in a dynamic equilibrium, with bone tissue constantly being renewed, thus maintaining bone hardness and elasticity. Excessive osteoclast activity disrupts this balance, leading to bone destruction in various diseases such as bone metastases from cancer, osteoporosis, rheumatoid arthritis, periodontitis, tooth loss, Iaget's disease, rickets, giant cell tumor of bone, and myeloma bone disease. (Chunhao Yang et al., Eur. J. Med. Chem. 2016, 123: 769-776).

[0004] Clinically, there are various drugs available for treating bone metabolic diseases that inhibit osteoclast formation. However, the efficacy of most drugs remains limited, and some have significant side effects and adverse reactions. For example, bisphosphonates can cause osteonecrosis of the frontal bone and inhibit bone formation; estrogen and selective estrogen receptor modulators are mainly targeted at postmenopausal women and carry an increased risk of deep vein thrombosis; calcitonin analogs may cause allergic reactions. Therefore, finding more effective and specific osteoclast inhibitors for treating diseases caused by osteoclast abnormalities is of great significance. Summary of the Invention

[0005] Specifically, this invention relates to a class of 3-amide-β-carboline compounds represented by the general formula P, their optical isomers, pharmaceutically usable salts and hydrates, their preparation methods, and their application in the preparation of medicaments for the prevention and treatment of diseases caused by abnormal osteoclast activity.

[0006] This invention provides, in one aspect, a 3-amide-β-carbamoline compound represented by the general formula P, or its optical isomers, pharmaceutically usable salts, and hydrates.

[0007]

[0008] in:

[0009] R1 represents 1 to 4 substituents, which are independently selected from hydrogen, halogen, C1-C4 alkyl, C1-C6 alkoxy, hydroxyC1-C6 alkyl, and C1-C6 alkylamine.

[0010] R2 and R3 are each independently hydrogen, substituted or unsubstituted C6-C. 20 Aryl, substituted or unsubstituted 5-10 membered heteroaryl, substituted or unsubstituted 3-10 membered heterocyclic, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C3-C 10 Cycloalkyl, substituted or unsubstituted C1-C 10 Alkoxy, or substituted or unsubstituted C1-C 10 Alkylamine group; or R2, R3 together with the N atom attached to them to form a substituted or unsubstituted 5-10 membered heteroaryl group, or a substituted or unsubstituted 3-10 membered heterocyclic group; preferably one of R2 and R3 is hydrogen;

[0011] n is an integer from 0 to 2, for example 0, 1 or 2, preferably 0 or 1;

[0012] R4 is hydrogen;

[0013] X is selected from CHR5R6, NR5R6, OR5, or SR5;

[0014] Alternatively, R4 and X can be connected to form -CH2-CHR5-, -CH2-NR5-, -CH2-O-, -CH2-S-;

[0015] R5 and R6 are each independently hydrogen-containing, substituted, or unsubstituted C6-C each time they appear. 20 Aryl, substituted or unsubstituted 5-20 membered heteroaryl; preferably, R6 is hydrogen;

[0016] The substitution in "substituted or non-substituted" above refers to substitution by a substituent selected from the following: halogen, hydroxyl, amino (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C6 alkyl, C1-C6 haloalkyl (e.g., fluoroalkyl, e.g., trifluoromethyl), C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyC1-C6 alkyl, benzyloxy, methanesulfonylimide acyl,

[0017] Ring A is C6-C 20 Aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclic;

[0018] R9 represents 1 to 6 substituents, which are independently selected from halogens, hydroxyl groups, amino groups (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyC1-C6 alkyl, benzyloxy, and methanesulfonylimide acyl.

[0019] R7 and R8 are each independently hydrogen and C1-C20 alkyl groups each time they appear.

[0020] Preferably,

[0021] R1 represents 1 to 4 substituents, which are independently selected from hydrogen, halogen, and methoxy groups;

[0022] R2 and R3 are each independently hydrogen, substituted or unsubstituted C6-C. 10 Aryl, substituted or unsubstituted 5-10-membered heteroaryl, substituted or unsubstituted 3-10-membered heterocyclic group, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C6 alkoxy, or substituted or unsubstituted C1-C6 alkylamine; or R2, R3 and the N atom attached to them together form a substituted or unsubstituted 5-10-membered heteroaryl or a substituted or unsubstituted 3-10-membered heterocyclic group; preferably one of R2 and R3 is hydrogen;

[0023] n is an integer from 0 to 2, for example 0, 1 or 2, preferably 0 or 1;

[0024] R4 is hydrogen;

[0025] X is selected from CHR5R6, NR5R6, OR5, or SR5;

[0026] Alternatively, R4 and X can be connected to form -CH2-CHR5-, -CH2-NR5-, -CH2-O-, -CH2-S-;

[0027] R5 and R6 are each independently hydrogen-containing, substituted, or unsubstituted C6-C each time they appear. 10 Aryl, substituted or unsubstituted 5-10 membered heteroaryl; preferably, R6 is hydrogen;

[0028] The substitution in "substituted or non-substituted" above refers to substitution by a substituent selected from the following: halogen, hydroxyl, amino (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C6 alkyl, C1-C6 haloalkyl (e.g., fluoroalkyl, e.g., trifluoromethyl), C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyC1-C6 alkyl, benzyloxy, methanesulfonylimide acyl,

[0029] Ring A is C6-C 10 Aryl, 5-10 membered heteroaryl, 3-10 membered heterocyclic;

[0030] R9 represents 1 to 6 substituents, which are independently selected from halogens, hydroxyl groups, amino groups (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C6 alkyl, C1-C6 haloalkyl, C3-C6 cycloalkyl, C1-C6 alkoxy, hydroxyC1-C6 alkyl, benzyloxy, and methanesulfonylimide acyl.

[0031] R7 and R8 are each independently hydrogen and C1-C10 alkyl groups each time they appear.

[0032] More preferably,

[0033] R1 represents 1 to 4 substituents, which are independently selected from hydrogen, halogen, and methoxy groups;

[0034] R2 and R3 are each independently hydrogen, substituted or unsubstituted C6-C. 10Aryl, substituted or unsubstituted 5-8-membered heteroaryl, substituted or unsubstituted 3-8-membered heterocyclic group, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted C1-C3 alkoxy, or substituted or unsubstituted C1-C3 alkylamine; or R2, R3 and the N atom attached to them together form a substituted or unsubstituted 5-8-membered heteroaryl, or a substituted or unsubstituted 3-8-membered heterocyclic group; preferably one of R2 and R3 is hydrogen;

[0035] n is an integer from 0 to 2, for example 0, 1 or 2, preferably 0 or 1;

[0036] R4 is hydrogen;

[0037] X is selected from CHR5R6, NR5R6, OR5, or SR5;

[0038] Alternatively, R4 and X can be connected to form -CH2-CHR5-, -CH2-NR5-, -CH2-O-, -CH2-S-;

[0039] R5 and R6 are each independently hydrogen-containing, substituted, or unsubstituted C6-C each time they appear. 10 Aryl, substituted or unsubstituted 5-8 membered heteroaryl; preferably, R6 is hydrogen;

[0040] The substitution in "substituted or non-substituted" above refers to substitution by a substituent selected from the following: halogen, hydroxyl, amino (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C6 alkyl, C1-C6 haloalkyl (e.g., fluoroalkyl, e.g., trifluoromethyl), C3-C6 cycloalkyl, C1-C3 alkoxy, hydroxyC1-C3 alkyl, benzyloxy, methanesulfonylimide acyl,

[0041] Ring A is C6-C 10 Aryl, 5-8 membered heteroaryl, 3-8 membered heterocyclic;

[0042] R9 represents 1 to 6 substituents, which are independently selected from halogens, hydroxyl groups, amino groups (-NH2), R7C(=O)NH-, R7S(=O)2NH-, NH2S(=O)2-, R7S(=O)-, R7S(=O)2-, R7R8P(=O)-, R7C(=O)-, cyano, C1-C3 alkyl, C1-C3 haloalkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, hydroxyC1-C3 alkyl, benzyloxy, and methanesulfonylimide acyl.

[0043] R7 and R8 are each independently hydrogen and C1-C6 alkyl groups each time they appear.

[0044] Preferably, the compound of general formula P is represented by the following general formulas P-1, P-2, or P-3:

[0045]

[0046] In the above general formulas P-1 to P-3, R1-R3, R5 and R6 are defined in the same way as described above.

[0047] Preferably, the compound of general formula P is represented by the following general formula P-3-1:

[0048]

[0049] In the general formulas P-1 to P-3 above, R1-R3, ring A, and R9 are defined as described above, and ring B is C6-C. 20 Aromatic rings, 5-20 yuan of mixed aromatic rings;

[0050] In this invention,

[0051] The term "alkyl" refers to an aliphatic hydrocarbon group, which can be branched or straight-chain alkyl. Depending on its structure, an alkyl group can be a monovalent or divalent group (i.e., an alkylene group), for example, in the case of a hydroxyl group (C0). 1-6 In "alkyl", the C 1-6 The alkyl group is actually a divalent group (alkylene group). In this invention, the alkyl group is preferably a "lower alkyl group" having 1-6 carbon atoms, and even more preferably a "lower alkyl group" having 1-3 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, etc.

[0052] The "halogens" mentioned are F, Cl, Br, and I;

[0053] The term "haloalkyl" refers to an alkyl group in which at least one hydrogen atom is replaced by a halogen atom. In some embodiments, if two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms may be the same or different from each other;

[0054] The term "heteroalkyl" refers to an alkyl group in which at least one skeletal carbon atom is replaced by a heteroatom (N, O, S). In some embodiments, if two or more carbon atoms are replaced by heteroatoms, the heteroatoms may be the same or different from each other.

[0055] The “cycloalkyl” is a saturated or unsaturated 3-10 member monocyclic or polycyclic alicyclic ring, and can be a monovalent group or a bivalent group (and a cycloalkylene ring);

[0056] The "heterocyclic group" is a saturated or unsaturated 3-10 member monocyclic or polycyclic alicyclic heterocycle containing one or more heteroatoms selected from N, O, and S. It can be a monovalent group or a bivalent group (i.e., a heterocyclic alkyl group).

[0057] The term "aryl" refers to an aromatic ring in which each constituent atom is a carbon atom, including monocyclic or fused polycyclic rings, and can be a monovalent or bivalent group (i.e., arylene). In this invention, the aryl ring preferably has 5-10 carbon atoms, and more preferably an aryl ring having 5-7 carbon atoms.

[0058] The term "aralkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by an aryl group. Examples include benzyl and phenethyl.

[0059] The term "heteroaryl" refers to an aromatic group containing one or more heteroatoms selected from N, O, and S on its ring. Depending on its structure, a heteroaryl can be a monovalent or bivalent group (i.e., a hypoaryl). Examples of heteroaryl groups include, but are not limited to, pyridinyl, pyridazolyl, pyrazolyl, pyrazinyl, triazolyl, tetrazolyl, thiophenyl, thiazolyl, furanyl, oxazolyl, isoxazolyl, pyrroleyl, quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl, indazoleyl, inazinyl, phthalazinyl, pyridazinyl, isoindolyl, pteridyl, purineyl, oxadiazolyl, thiazolyl, furazonyl, benzofuranyl, benzothiophenyl, benzothiazolyl, benzooxazolyl, quinazolinyl, etc.

[0060] The "heterocyclic group" is monocyclic or polycyclic, and at least one of them is a saturated or partially saturated (non-aromatic) ring having one or more heteroatoms. Representative heterocyclic groups include the following three cases: 1) each ring is a non-aromatic ring, and at least one ring has a heteroatom; 2) at least one ring is a non-aromatic ring containing a heteroatom, and at least one other ring is an aromatic carbon ring; 3) at least one ring is a non-aromatic ring containing a heteroatom, and at least one other ring is a heteroaromatic ring.

[0061] Most preferably, the 3-amido-β-carboline compound represented by general formula I is selected from the following compounds:

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070] In another aspect, this invention provides a method for preparing 3-amido-β-carboline compounds represented by the general formula Pa, Pb, or Pc, which can be obtained by the following four methods:

[0071] Preparation method 1:

[0072]

[0073] Step i: The starting material undergoes a condensation and cyclization reaction with acetaldehyde to give compound a1;

[0074] Step ii: Compound a1 is oxidatively aromatized with NCS (N-chlorosuccinimide) or DDQ (2,3-dichloro-5,6-dicyanobenzoquinone) to obtain β-carboline compound a2;

[0075] Step iii: Compound a2 is oxidized by m-CPBA (m-chloroperoxybenzoic acid) or hydrogen peroxide to obtain pyridine peroxide compound a3;

[0076] Step iv: Compound a3 undergoes a rearrangement reaction in the presence of a strong Lewis acid, such as trifluoroacetic acid, to give benzyl alcohol compound a4;

[0077] Step v: Compound a4 is hydrolyzed in the presence of a base, such as potassium hydroxide, sodium hydroxide, or lithium hydroxide, to give 3-carboxy-β-carboline compound a5;

[0078] Step vi: Compound a5 and 2,6-dimethylaniline are condensed with condensing agents such as HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) to form amide compound I1;

[0079] Step vii: Compound I1 is oxidized to aldehyde I2 by IBX (2-iodobenzoic acid) or Desmartin oxidant;

[0080] Step viii: Compound I1 is brominated to benzyl bromide product I3 under the action of phosphorus tribromide;

[0081] Step ix: Intermediate I2 is subjected to reductive amination, and intermediate I3 is subjected to substitution reaction to obtain a 3-amide-β-carboline compound represented by the general formula Pa.

[0082] Preparation method two:

[0083]

[0084] Step i: The starting material and 2,6-dimethylaniline are condensed in the presence of condensing agents such as HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) to form an amide compound a6;

[0085] Step ii: Compound a6 undergoes a deprotection reaction under the action of hydrogen chloride or trifluoroacetic acid to obtain the corresponding salt, which is then directly added to the next reaction step;

[0086] Step iii: The corresponding salt undergoes a condensation cyclization reaction with 1-Boc-N-heterocyclic butane-3-aldehyde to give intermediate a7;

[0087] Step iv: Compound a7 was oxidatively aromatized with NCS (N-chlorosuccinimide) or DDQ (2,3-dichloro-5,6-dicyanobenzoquinone) to give β-carboline compound a8;

[0088] Step v: Compound a8 undergoes a deprotection (-Boc) reaction under the action of hydrogen chloride or trifluoroacetic acid to give the corresponding base I4;

[0089] Step vi: Compound I4 undergoes a coupling reaction under the catalysis of transition metals such as copper and palladium to obtain 3-amide-β-carboline compounds represented by the general formula Pb.

[0090] Preparation method three:

[0091]

[0092] Step i: The starting material and organic amine are condensed in the presence of condensing agents such as HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) to form amide compound a9;

[0093] Step ii: Compound a9 undergoes a deprotection reaction under the action of hydrogen chloride or trifluoroacetic acid to obtain the corresponding salt, which is then directly added to the next reaction step;

[0094] Step iii: The corresponding salt undergoes a condensation cyclization reaction with substituted or unsubstituted arylpropionaldehyde to give intermediate a10;

[0095] Step iv: Compound a10 is oxidatively aromatized with NCS (N-chlorosuccinimide) or DDQ (2,3-dichloro-5,6-dicyanobenzoquinone) to obtain 3-amido-β-carboline compounds represented by general formula Pc;

[0096] Preparation method four:

[0097]

[0098] I5 undergoes a coupling reaction with organic amines (e.g., under the catalysis of transition metals such as copper and palladium) to yield 3-amide-β-carboline compounds represented by the general formula Pd.

[0099] In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the above-described 3-amido-β-carboline compounds represented by the general formula P, their optical isomers, pharmaceutically usable salts and hydrates, and pharmaceutically acceptable excipients.

[0100] Another aspect of the present invention provides the use of the 3-amido-β-carboline compound represented by the above general formula I, its optical isomers, pharmaceutically usable salts and hydrates, or the above pharmaceutical compositions in the preparation of a medicament for treating bone metabolism-related diseases (such as RANK-RANKL pathway-related diseases).

[0101] The bone metabolic diseases mentioned include osteoporosis, rheumatoid arthritis, psoriatic arthritis, bone metastasis of solid tumors, and tumors such as breast cancer.

[0102] The 3-amido-β-carboline compounds, their optical isomers, pharmaceutically usable salts, and hydrates of this invention can intervene in the OPG-RANKL-RANK signaling system by inhibiting RANKL-RANK interactions, thereby regulating RANKL activity in osteoclast precursor cells and inhibiting osteoclast formation, thus reducing bone resorption. The compounds, their optical isomers, pharmaceutically usable salts, and hydrates of this invention hold promise for the prevention and treatment of bone metabolic diseases. Detailed Implementation

[0103] The present invention will be further described below with reference to embodiments, but these embodiments are by no means intended to limit the present invention. In all embodiments, Bruker Avance III 600, Bruker Avance III 500 and Varian-Mercury Ilus-400 nuclear magnetic resonance spectrometers were used for recording, and chemical shifts were expressed as δ (ppm); the silica gel used for separation was 200-300 mesh.

[0104] Preparation Example 1: Synthesis of compound a1:

[0105]

[0106] 10.0 g (39.26 mmol) of tryptophan methyl ester hydrochloride was dissolved in 50 mL of acetonitrile. 1 mL (9 mmol) of trifluoroacetic acid was added, and 2.6 mL (47.12 mmol) of acetaldehyde was added under ice bath conditions. After the addition was complete, the ice bath was removed, and the mixture was allowed to react overnight at room temperature. The reaction solution was concentrated by rotary evaporation, and the reaction was quenched by adding saturated sodium carbonate solution. The pH was adjusted to 10-12, and the mixture was extracted four times with dichloromethane (50 mL × 4). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain crude oily liquid a1, which could be directly used for the next reaction without further purification.

[0107] Preparation Example 2: Synthesis of compound a2:

[0108]

[0109] Crude product a1 (3.6 g, 15 mmol) was dissolved in 20 mL of DMF, and 5 mL (31.5 mmol) of triethylamine was added. 4.2 g (31.5 mmol) of NCS was added under ice bath conditions. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 1 to 2 hours. The reaction was monitored by TLC. After the reaction was complete, 60 mL of saturated ammonium chloride solution was added to quench the reaction, and the product was extracted three times with ethyl acetate (20 mL × 3). The organic phases were combined, washed three times with saturated brine (40 mL × 3), and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain an oily crude product, which was purified by silica gel column chromatography to obtain the pale yellow target product a2 (42%). 1 ¹H NMR (400MHz, chloroform-d) δ 9.00 (s, 1H), 8.80 (s, 1H), 8.17 (d, J = 7.9Hz, 1H), 7.58 (d, J = 3.7Hz, 2H), 7.35 (ddd, J = 8.1, 4.8, 3.3Hz, 1H), 4.04 (s, 3H), 2.86 (s, 3H). MS (ESI) m / z: [M+1] +

[0110] Preparation Example 3: Synthesis of compound a3:

[0111]

[0112] Add a2 (5 g, 20.8 mmol) and 10.8 g (62.4 mmol) m-CPBA to an ice bath, dissolve in 50 mL of chloroform, bring to room temperature, and heat under reflux overnight. After the reaction is complete on TLC, concentrate the reaction solution by rotary evaporation to obtain the crude product, which is then purified by silica gel column chromatography to obtain the white target product a3 (86%). 1¹H NMR (400MHz, methanol-d⁴) δ 8.34 (s, 1H), 8.10 (dt, J = 7.9, 1.1 Hz, 1H), 7.56 (dd, J = 3.7, 1.2 Hz, 2H), 7.31 (ddd, J = 8.0, 5.0, 3.0 Hz, 1H), 4.02 (s, 3H), 2.79 (s, 3H).

[0113] Preparation Example 4: Synthesis of compound a4:

[0114]

[0115] Compound a3 (1.6 g, 6.25 mmol) was dissolved in 30 mL of dichloromethane. 4.3 mL (31.25 mmol) of trifluoroacetic anhydride was added under ice bath conditions. After the addition was complete, the ice bath was removed, and the mixture was brought to room temperature and refluxed overnight. After the reaction was complete on TLC, the reaction solution was concentrated by rotary evaporation to obtain a crude product. This crude product was then purified by silica gel column chromatography to obtain the white target product a4 (90%). 1 ¹H NMR (400MHz, methanol-d⁴) δ 8.92 (s, 1H), 8.30 (d, J = 8.1 Hz, 1H), 7.70 (d, J = 6.4 Hz, 2H), 7.40 (ddd, J = 8.1, 5.9, 2.1 Hz, 1H), 5.25 (s, 2H), 4.07 (s, 3H).

[0116] Preparation Example 5: Synthesis of Compound a5:

[0117]

[0118] Compound a4 (1.6 g, 6.25 mmol) was dissolved in 25 mL of methanol, 5 mL of water was added, and then 788 mg (18.75 mmol) of lithium hydroxide monohydrate was added. The mixture was stirred overnight at room temperature. After the reaction was confirmed to be complete by TLC, the methanol was distilled off, 10 mL of water was added, and the pH was adjusted to 1 by adding 1N hydrochloric acid solution dropwise with stirring in an ice bath. A large amount of solid precipitated out. The solid was filtered, and the filter cake was dried to obtain the yellow crude compound a5, which did not require further processing and was directly added to the next step. 1 H NMR (400MHz, DMSO-d6) δ12.13(s,1H),8.92(s,1H),8.42(d,J=7.9Hz,1H),7.7 4(d,J=8.3Hz,1H),7.62(t,J=7.7Hz,1H),7.33(t,J=7.5Hz,1H),5.08(s,2H).

[0119] Preparation Example 6: Synthesis of Intermediate I1:

[0120]

[0121] Compound a5 (0.371 g, 1.53 mmol) was dissolved in DMF, and diisopropylethylamine (DIPEA) (0.75 mL, 4.59 mmol) was added, followed by HATU (0.875 g, 2.3 mmol). The mixture was stirred at room temperature until homogeneous, and 2,6-dimethylaniline (0.28 mL, 2.3 mmol) was added. The mixture was heated to 80°C overnight. After the reaction was monitored by TLC to ensure complete reaction, saturated ammonium chloride solution was added to quench the reaction. The mixture was extracted three times with ethyl acetate, and the organic phases were combined and washed three times with saturated brine. The organic phase was separated and dried over anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography to obtain the yellow target product I1 (71%).

[0122] Preparation Example 7: Synthesis of Intermediate I2:

[0123]

[0124] Compound I1 (200 mg, 0.58 mmol) was dissolved in dichloromethane, and Dysmartin oxidant (297 mg, 0.7 mmol) was added. The reaction was carried out at room temperature for 1 hour, and the reaction was monitored by TLC. After the reaction was complete, sodium thiosulfate was added to quench the reaction. After three extractions with dichloromethane, the organic phases were combined and washed three times with saturated sodium bicarbonate aqueous solution. The organic phase was separated and dried with anhydrous sodium sulfate. The organic phase was concentrated and purified by silica gel column chromatography to obtain the pale yellow target product I2 (78%).

[0125] Preparation Example 8: Synthesis of Intermediate I3:

[0126]

[0127] I1 (0.1 g, 0.29 mmol) was dissolved in DMF, and phosphorus tribromide (0.236 g, 0.87 mmol) was added under ice bath conditions. After the addition was complete, the ice bath was removed, and the reaction was allowed to proceed overnight at room temperature. After the reaction was monitored by TLC to confirm its completeness, saturated sodium bromide was added to quench the reaction. The mixture was then extracted three times with ethyl acetate. The combined organic phases were washed three times with saturated sodium bromide solution, separated, and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain the crude compound I3, which, without further purification, was directly introduced to the next step.

[0128] Preparation of Example 9: General Steps A:

[0129]

[0130] To a 1,2-dichloroethane solution of intermediate I2 (0.23 mmol), the corresponding substrate amine (0.25 mmol) was added, followed by a catalytic amount of acetic acid (5%). After reacting at room temperature for one hour, sodium triacetoxyborohydride (0.35 mmol) was added, and the mixture was allowed to react overnight at room temperature. The reaction was monitored by TLC. After the starting material had reacted completely, a saturated sodium bicarbonate solution was added to quench the reaction. The mixture was then extracted three times with dichloromethane. The organic phases were combined and dried over anhydrous sodium sulfate. The resulting product was purified by silica gel column chromatography to obtain the target product.

[0131] Preparation of Example 10: General Step B:

[0132]

[0133] Under nitrogen protection, ultra-dry DMF and the corresponding substrate amine (0.44 mmol) were added to intermediate I2 (118 mg, 0.29 mmol), followed by potassium iodide (0.15 mmol) and cesium carbonate (0.44 mmol). The mixture was heated to 80°C and reacted for two hours, monitored by TLC. After the starting material was completely consumed, heating was stopped, and the mixture was cooled to room temperature. The reaction was quenched with saturated brine, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed three times with saturated brine, separated, and dried over anhydrous sodium sulfate. The resulting product was purified by silica gel column chromatography.

[0134] Example 1: N-(2,6-dimethylphenyl)-1-((aniline)methyl)-9H-pyrido[3,4-b]indole-3-amide (P1)

[0135]

[0136] The synthesis method is described in general step A, using aniline as the substrate amine. White solid (51%). 1 H NMR (400MHz, DMSO-d6) δ12.22(d,J=3.4Hz,1H),10.46(d,J=3.3Hz,1H),8.88(d,J=3.7Hz,1H),8.42(dd,J=8.1,3.6Hz,1H),7.71(dd,J=8.2,3. 7Hz,1H),7.63(q,J=8.3,6.3Hz,1H),7.34(dd,J=7.7,4.0Hz,1H),7.30–6.99(m,8H),6.82(s,1H),4.98(s,2H),2.26(d,J=3.8Hz,6H).LRMS(ESI + m / z:420.9[M+1] + .

[0137] Example 2: N-(2,6-dimethylphenyl)-1-((methyl(phenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P2)

[0138]

[0139] The synthesis method is described in general step B, using N-methylaniline as the substrate amine. White solid (64%). 1 H NMR (600MHz, DMSO-d6) δ12.08(s,1H),9.30(s,1H),8.79(s,1H),8.41(d,J=7.8Hz,1H),7.73(d,J=8.3Hz,1H),7.66–7.60(m,1H),7.3 2(t,J=7.5Hz,1H),7.11–7.05(m,5H),6.89(d,J=8.2Hz,2H),6.53(t,J=7.2Hz,1H),5.18(s,2H),3.21(s,3H),2.06(s,6H).LRMS(ESI + m / z:434.8[M+1] + .

[0140] Example 3: N-(2,6-dimethylphenyl)-1-(((4-fluorophenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P3)

[0141]

[0142] The synthesis method is described in general step A, using 4-fluoroaniline as the corresponding substrate amine. White solid (45%). 1 HNMR (400MHz, DMSO-d6) δ12.30(s,1H),10.49(s,1H),8.89(s,1H),8.42(d,J=8.0Hz,1H),7.70(d,J=8.2 Hz,1H),7.67–7.60(m,1H),7.32(t,J=7.4Hz,1H),7.29–7.07(m,7H),5.00(s,2H),2.25(s,6H).LRMS(ESI + m / z:438.8[M+1] + .

[0143] Example 4: N-(2,6-dimethoxyphenyl)-1-(((2-fluorophenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P4)

[0144]

[0145] The synthesis method is described in general step A, using 2-fluoroaniline as the corresponding substrate amine. White solid (39%). 1 HNMR (400MHz, DMSO-d6) δ12.45(s,1H),10.31(s,1H),8.86(s,1H),8.40(d,J=7.9Hz,1H),7.72(d,J=8.3Hz,1H),7.62(t,J=7.5Hz,1H),7.3 2(t,J=7.5Hz,1H),7.23–7.12(m,4H),7.02–6.96(m,1H),6.89(t,J=7.7Hz,1H),6.54(q,J=6.9Hz,1H),4.96(s,2H),2.23(s,6H).LRMS(ESI + m / z:438.8[M+1] + .

[0146] Example 5: 1-(((2,4-difluorophenyl)amino)methyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P5)

[0147]

[0148] The synthesis method is described in general step A, using 2,4-difluoroaniline as the corresponding substrate amine. White solid (22%). 1 HNMR (400MHz, DMSO-d6) δ12.37(s,1H),10.28(s,1H),8.83(s,1H),8.40(d,J=8.0Hz,1H),7.71(d,J=8.3Hz,1H),7.62(t,J=7.7Hz ,1H),7.31(t,J=7.5Hz,1H),7.20–7.11(m,4H),7.06(t,J=10.4Hz,1H),6.77(t,J=9.2Hz,1H),4.92(s,2H),2.23(s,6H).LRMS(ESI + m / z: 456.7 [M+1] + .

[0149] Example 6: 1-(((2,6-difluorophenyl)amino)methyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P6)

[0150]

[0151] The synthesis method is described in general step B, using 2,6-difluoroaniline as the corresponding substrate amine. White solid (12%).1 HNMR (400MHz, DMSO-d6) δ12.14(s,1H),9.74(s,1H),8.79(s,1H),8.40(d,J=7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.66–7.59(m,1H),7.31(t,J =7.4Hz,1H),7.15(s,3H),6.90–6.77(m,2H),6.59(ddd,J=14.6,8.3,6.0Hz,1H),6.13(s,1H),5.02(d,J=6.7Hz,2H),2.17(s,6H).LRMS(ESI + m / z: 456.9 [M+1] + .

[0152] Example 7: N-(2,6-dimethylphenyl)-1-(((2-(methylsulfonyl)phenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P7)

[0153]

[0154] The synthesis method is described in general step A, using 2-methanesulfonylaniline as the corresponding substrate amine. White solid (54%). 1 H NMR(500MHz,DMSO-d6)δ13.01–12.78(m,1H),9.88(s,1H),8.93(s,1H),8.43 (dd,J=7.9,2.4Hz,1H),8.24–7.80(m,1H),7.73(d,J=8.2Hz,1H),7.66–7.60( m,2H),7.58(tt,J=8.4,1.6Hz,1H),7.32(td,J=7.6,2.0Hz,1H),7.18–7.14(m ,4H),6.82(t,J=7.5Hz,1H),5.08(s,2H),3.13(s,3H),2.23(s,6H).LRMS(ESI + )m / z:498.8[M+1] + .

[0155] Example 8: N-(2,6-dimethylphenyl)-1-(((4-(methylsulfonyl)phenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P8)

[0156]

[0157] The synthesis method is described in general step A, using 4-methanesulfonylaniline as the corresponding substrate amine. White solid (50%).1 H NMR (400MHz, DMSO-d6) δ12.31(s,1H),10.38(s,1H),8.89(s,1H),8.42(d,J=7.9Hz,1H),7.72(d,J=8.1Hz,1H),7.67–7 .60(m,3H),7.33(t,J=7.5Hz,1H),7.17(s,3H),7.04(d,J=8.7Hz,2H),4.97(s,2H),3.02(s,3H),2.24(s,6H).LRMS(ESI + m / z:498.7[M+1] + .

[0158] Example 9: N-(2,6-dimethylphenyl)-1-(((4-(methanesulfonyl)phenyl)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P9)

[0159]

[0160] The synthesis method is described in general step A, using 4-methanesulfonamide aniline as the corresponding substrate amine. White solid (63%). 1 H NMR(400MHz,DMSO-d6)δ12.10(s,1H),10.40(s,1H),9.00(s,1H),8.85(s,1H ),8.41(d,J=7.9Hz,1H),7.71(d,J=8.2Hz,1H),7.62(t,J=7.7Hz,1H),7.32( t,J=7.5Hz,1H),7.17(s,3H),7.06–6.99(m,2H),6.88(d,J=8.6Hz,2H),6.62 (t,J=5.5Hz,1H),4.83(d,J=5.3Hz,2H),2.80(s,3H),2.24(s,6H).LRMS(ESI + m / z: 513.7 [M+1] + .

[0161] Example 10: N-(2,6-dimethylphenyl)-1-((2-aminopyridyl)methyl)-9H-pyrido[3,4-b]indole-3-amide (P10)

[0162]

[0163] The synthesis method is described in general step A, using pyridine-2-amine as the corresponding substrate amine. White solid (21%). 1HNMR(500MHz,DMSO-d6)δ12.68–12.61(m,1H),10.32(s,1H),9.83–9.73(m,1H),8.93(s,1H),8.43(d,J=7.8Hz,1H),8.08(d,J=9.0Hz,2H),7.7 2(d,J=8.3Hz,1H),7.64(t,J=7.6Hz,1H),7.33(t,J=7.9Hz,2H),7.12(s,3H),6.94(t,J=6.6Hz,1H),5.36–5.24(m,2H),2.20(s,6H).LRMS(ESI + m / z:421.8[M+1] + .

[0164] Example 11: N-(2,6-dimethylphenyl)-1-((3-aminopyridyl)methyl)-9H-pyrido[3,4-b]indole-3-amide (P11)

[0165]

[0166] The synthesis method is described in general step A, using pyridine-3-amine as the corresponding substrate amine. White solid (27%). 1 HNMR(500MHz,DMSO-d6)δ13.02(s,1H),10.54(s,1H),8.87(s,1H),8.49(d,J=2 .7Hz,1H),8.39(d,J=7.8Hz,1H),8.13(dd,J=8.8,2.6Hz,1H),8.04(d,J=5.3Hz, 1H),7.77(dd,J=8.8,5.3Hz,1H),7.70(d,J=8.2Hz,1H),7.61(ddd,J=8.3,7.0, 1.2Hz,1H),7.32–7.27(m,1H),7.14(s,3H),5.10(s,2H),2.22(s,6H).LRMS(ESI + m / z:421.8[M+1] + .

[0167] Example 12: N-(2,6-dimethylphenyl)-1-((4-aminopyridyl)methyl)-9H-pyrido[3,4-b]indole-3-amide (P12)

[0168]

[0169] The synthesis method is described in general step B, using pyridine-4-amine as the corresponding substrate amine. White solid (16%). 1HNMR(400MHz,Chloroform-d)δ9.77–9.70(m,1H),8.93(s,1H),8.73–8.62(m,2H),8.45(d,J=8.0Hz,1H),8.21(d,J=7.7Hz,2H),7.75(d,J= 8.2Hz,1H),7.66(t,J=7.7Hz,1H),7.35(t,J=7.5Hz,1H),7.16(s,3H),6.87(d,J=6.8Hz,2H),5.99(t,J=4.9Hz,2H),2.18(s,6H).LRMS(ESI) + m / z:421.8[M+1] + .

[0170] Example 13: N-(2,6-dimethylphenyl)-1-((2-aminothiazolyl)methyl)-9H-pyrido[3,4-b]indole-3-amide (P13)

[0171]

[0172] The synthesis method is described in general step A, using thiazole-2-amine as the corresponding substrate amine. White solid (39%). 1 HNMR (400MHz, DMSO-d6) δ12.35(s,1H),10.25(s,1H),8.93(s,1H),8.45(d,J=7.9Hz,1H),7.72(d,J=8.3Hz,1H),7.64(t,J =7.7Hz,1H),7.47–7.40(m,1H),7.34(t,J=7.7Hz,1H),7.16(s,3H),7.04–6.97(m,1H),5.24(s,2H),2.22(s,6H).LRMS(ESI + m / z:427.8[M+1] + .

[0173] Example 14: N-(2,6-dimethylphenyl)-1-(((1-methyl-1H-pyrazole-4-substituted)amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P14)

[0174]

[0175] The synthesis method is described in general step A, using 1-methyl-1H-pyrazole-4-amine as the corresponding substrate amine. White solid (43%). 1H NMR (400MHz, DMSO-d6) δ12.66–12.44(m,1H),12.16(s,2H),10.70(s,1H),8.94(s,1H),8.45(d,J=8.0Hz,1H),8.0 9(s,1H),7.72–7.59(m,3H),7.33(t,J=7.4Hz,1H),7.17(s,3H),5.18(s,2H),3.81(s,3H),2.26(s,6H).LRMS(ESI + m / z:424.8[M+1] + .

[0176] Example 15: 1-(((1H-imidazol-2-substituted)amino)methyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P15)

[0177]

[0178] The synthesis method is described in general step B, using 1H-imidazol-2-amine as the corresponding substrate amine. White solid (15%). 1 H NMR (400MHz, DMSO-d6) δ12.51(s,1H),12.02(s,1H),9.65(s,1H),8.92(s,1H),8.46(d,J=8.0Hz,1H),7.75(d,J=8.3Hz,1H),7.66(ddd, LRMS(ESI) + m / z:410.8[M+1] + .

[0179] Example 16: N-(2,6-dimethylphenyl)-1-((isoxazo-4-substituted amino)methyl)-9H-pyrido[3,4-b]indole-3-amide (P16)

[0180]

[0181] The synthesis method is described in general step A, using isoxazol-4-amine as the corresponding substrate amine. White solid (27%). 1HNMR (400MHz, DMSO-d6) δ12.35(s,1H),10.41(s,1H),8.87(s,1H),8.50(d,J=5.4Hz,2H),8.41(d,J=7.9Hz,1H),7. 70(d,J=8.2Hz,1H),7.62(t,J=7.7Hz,1H),7.32(t,J=7.4Hz,1H),7.17(s,3H),4.78(s,2H),2.23(s,6H).LRMS(ESI + m / z:411.8[M+1] + .

[0182] Example 17: 1-(((1H-pyrazole-3-substituted)amino)methyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P17)

[0183]

[0184] The synthesis method is described in general step A, using 1H-pyrazole-3-amine as the corresponding substrate amine. White solid (50%). 1 H NMR (400MHz, DMSO-d6) δ12.41(s,1H),10.23(s,1H),8.89(s,1H),8.43(d,J=7.9Hz,1H),7.98(d,J=2.8Hz,1H),7.71(d,J=8 .2Hz,1H),7.63(t,J=7.6Hz,1H),7.33(t,J=7.5Hz,1H),7.16(s,3H),6.13–6.05(m,1H),5.06(s,2H),2.21(s,6H).LRMS(ESI + m / z:410.8[M+1] + .

[0185] Preparation Example 11: Synthesis of Intermediate I4:

[0186]

[0187] Dissolve a6 (4 g, 9.8 mmol) in 36 mL of dichloromethane, add trifluoroacetic acid (7.3 mL, 98 mmol) under ice bath, stir overnight at room temperature, and monitor the reaction by TLC. After the reaction is complete, concentrate the reaction solution to obtain crude trifluoroacetate. Dissolve the crude product in acetonitrile, add substrate a7 (2.8 g, 21 mmol), and react overnight. After the reaction is complete by TLC, concentrate the reaction solution by rotary evaporation, quench the reaction with saturated sodium carbonate solution, adjust the pH to 10-12, extract four times with dichloromethane (50 mL × 4), combine the organic phases, dry with anhydrous sodium sulfate, concentrate the organic phase to obtain an oily liquid crude product, dissolve the crude product in DMF, add triethylamine (3.5 mL, 25 mmol), add NCS (2.8 g, 21 mmol) under ice bath, remove the ice bath, and react at room temperature for 1 to 2 hours, monitoring the reaction by TLC. After the reaction was complete, 60 mL of saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (20 mL × 3). The organic phases were combined, washed three times with saturated brine (40 mL × 3), and dried with anhydrous sodium sulfate. The organic phase was concentrated to obtain an oily liquid crude product, which was then purified by silica gel column chromatography to obtain the pale yellow target product (60%).

[0188]

[0189] 4M hydrochloric acid-methanol solution was added to a8 (4.7 g, 10 mmol) dissolved in methanol. After reacting at room temperature for two hours, the reaction was monitored by TLC. The methanol was removed to obtain crude I4 hydrochloride. 1N sodium hydroxide aqueous solution was added to adjust the pH to 14. The solution was extracted five times with dichloromethane until no target product was found in the aqueous phase. The organic phases were combined and dried with anhydrous sodium sulfate. The organic phase was concentrated to obtain crude I4 (75%) as a yellow solid. 1 H NMR(400MHz, Methanol-d4)δ8.94(s,1H),8.31–8.27(m,1H),7.70–7.62(m,2H),7.36(ddd,J=8.0, 6.8,1.3Hz,1H),7.18(s,3H),4.95–4.90(m,1H),4.71(td,J=11.8,11.2,8.2Hz,4H),2.36(s,6H).

[0190] Preparation Example 12: General Step C: Synthesis of Target Products P18-P54

[0191]

[0192] Under argon protection, I4 (93 mg, 0.25 mmol), aryl bromide (0.5 mmol), Pd2(dba)3 (0.025 mmol), RuPhos (0.025 mmol), and cesium carbonate (244 mg, 0.75 mmol) were added, followed by reflux with toluene overnight. After TLC monitoring to confirm complete reaction, heating was stopped, and the reaction was allowed to cool to room temperature. The reaction was then quenched with saturated ammonium chloride solution, followed by extraction once with ethyl acetate, and then four times with dichloromethane until complete extraction. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain an oily crude product, which was purified by silica gel column chromatography. The purified solid was recrystallized from ethyl acetate, cooled to room temperature, filtered, and dried to obtain a white solid target compound.

[0193] Example 18: N-(2,6-dimethylphenyl)-1-(1-phenylazacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P18)

[0194]

[0195] The synthesis method follows general procedure C, using bromobenzene as the corresponding aryl bromide. White solid (77%). 1 HNMR(500MHz,DMSO-d6)δ12.00(s,1H),9.97(s,1H),8.83(s,1H),8.41(d,J= 7.8Hz,1H),7.69(d,J=8.2Hz,1H),7.62(ddd,J=8.3,7.0,1.2Hz,1H),7.32(t ,J=7.5Hz,1H),7.21–7.14(m,2H),7.12(s,3H),6.67(t,J=7.3Hz,1H),6.58– 6.52(m,2H),4.75–4.66(m,1H),4.41(p,J=7.1Hz,4H),2.18(s,6H).LRMS(ESI + m / z:447.3[M+1] + .

[0196] Example 19: N-(2,6-dimethylphenyl)-1-(1-(2-fluorophenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P19)

[0197]

[0198] The synthesis method follows general procedure C, using 2-fluoro-1-bromobenzene as the corresponding aryl bromide. White solid (61%). 1H NMR (400MHz, DMSO-d6) δ11.99(s,1H),10.04(s,1H),8.83(s,1H),8.41(d,J=7.8Hz,1H ),7.69(d,J=8.2Hz,1H),7.62(ddd,J=8.2,6.9,1.2Hz,1H),7.35–7.27(m,1H),7.13(s, 3H),7.06(dd,J=8.5,1.4Hz,1H),7.02(ddd,J=7.8,5.9,1.7Hz,1H),6.75–6.67(m,2H), 4.72(p,J=7.4Hz,1H),4.54(t,J=6.8Hz,2H),4.52–4.46(m,2H),2.20(s,6H).LRMS(ESI + m / z: 464.9 [M+1] + .

[0199] Example 20: N-(2,6-dimethylphenyl)-1-(1-(3-fluorophenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P20)

[0200]

[0201] The synthesis method follows general procedure C, using 3-fluoro-1-bromobenzene as the corresponding aryl bromide. White solid (80%). 1 H NMR (400MHz, Chloroform-d) δ10.42(s,1H),9.60(s,1H),8.99(s,1H),8.21(d,J=7. 9Hz,1H),7.56(dt,J=17.8,8.0Hz,2H),7.35(t,J=7.4Hz,1H),7.30(dd,J=8.2,6.6H z,1H),7.14(s,3H),6.66–6.60(m,1H),6.49(dd,J=8.1,2.1Hz,1H),6.41(dt,J=10. 8,2.1Hz,1H),4.49–4.39(m,4H),4.33(dq,J=8.6,3.9Hz,1H),2.34(s,6H).LRMS(ESI + m / z: 464.8 [M+1] + .

[0202] Example 21: N-(2,6-dimethylphenyl)-1-(1-(4-fluorophenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P21)

[0203]

[0204] The synthesis method follows general procedure C, using 4-fluoro-1-bromobenzene as the corresponding aryl bromide. White solid (75%). 1 H NMR(600MHz,DMSO-d6)δ12.01(s,1H),9.98(s,1H),8.84(s,1H),8.41(s,1H),7.76–7.57(m,2H),7 .31(s,1H),7.18–6.95(m,5H),6.55(s,2H),4.69(s,1H),4.50–4.27(m,4H),2.19(s,6H).LRMS(ESI + m / z: 465.3 [M+1] + .

[0205] Example 22: 1-(1-(2-chlorophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P22)

[0206]

[0207] The synthesis method follows general procedure C, using 2-chloro-1-bromobenzene as the corresponding aryl bromide. White solid (71%). 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.04(s,1H),8.84(s,1H),8.41(d,J=7.9Hz,1H ),7.69(d,J=8.2Hz,1H),7.62(ddd,J=8.2,7.0,1.2Hz,1H),7.32(t,J=7.3Hz,1H),7.25 (dd,J=7.9,1.5Hz,1H),7.21–7.16(m,1H),7.13(s,3H),6.81(dd,J=8.2,1.5Hz,1H),6. 76(td,J=7.6,1.5Hz,1H),4.71–4.63(m,1H),4.61–4.57(m,4H),2.20(s,6H).LRMS(ESI + m / z: 480.9 [M+1] + .

[0208] Example 23: 1-(1-(3-chlorophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P23)

[0209]

[0210] The synthesis method follows general procedure C, using 3-chloro-1-bromobenzene as the corresponding aryl bromide. White solid (70%). 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.01(s,1H),8.84(s,1H),8.41(d,J=7.9Hz,1H),7.69 (d,J=8.1Hz,1H),7.62(ddd,J=8.2,7.0,1.2Hz,1H),7.34–7.29(m,1H),7.17(t,J=8.0Hz,1H), 7.13(s,3H),6.66(dd,J=7.7,2.0Hz,1H),6.56(t,J=2.1Hz,1H),6.48(dd,J=8.1,2.2Hz,1H),4 .71(h,J=6.7,6.0Hz,1H),4.49(t,J=6.7Hz,2H),4.41(t,J=7.7Hz,2H),2.19(s,6H).LRMS(ESI + m / z: 480.9 [M+1] + .

[0211] Example 24: 1-(1-(4-chlorophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P24)

[0212]

[0213] The synthesis method follows general procedure C, using 4-chloro-1-bromobenzene as the corresponding aryl bromide. White solid (76%). 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),9.98(s,1H),8.84(s,1H),8.41(d,J=7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.63(dd,J=7.0,1.3Hz,1H),7.35–7.2 7(m,1H),7.22–7.17(m,2H),7.13(s,3H),6.58–6.53(m,2H),4.76–4.65(m ,1H),4.45(t,J=6.7Hz,2H),4.40(t,J=7.6Hz,2H),2.19(s,6H).LRMS(ESI + m / z: 480.8 [M+1] + .

[0214] Example 25: N-(2,6-dimethylphenyl)-1-(1-(o-methylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P25)

[0215]

[0216] The synthesis method follows general procedure C, using 2-methyl-1-bromobenzene as the corresponding aryl bromide. White solid (83%). 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),10.03(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1 H),7.69(d,J=8.2Hz,1H),7.62(t,J=7.6Hz,1H),7.31(t,J=7.4Hz,1H),7.13(s,3H),7 .06(d,J=7.7Hz,1H),7.00(d,J=7.4Hz,1H),6.69(t,J=7.4Hz,1H),6.63(d,J=8.0Hz, 1H),4.65(p,J=7.5Hz,1H),4.46(p,J=7.1Hz,4H),2.24(s,3H),2.20(s,6H).LRMS(ESI + )m / z:461.0[M+1] + .

[0217] Example 26: N-(2,6-dimethylphenyl)-1-(1-(m-methylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P26)

[0218]

[0219] The synthesis method follows general procedure C, using 3-methyl-1-bromobenzene as the corresponding aryl bromide. White solid (86%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.97(s,1H),8.84(s,1H),8.41(d,J=7. 9Hz,1H),7.69(d,J=8.2Hz,1H),7.62(t,J=7.6Hz,1H),7.32(t,J=7.5Hz,1H), 7.13(s,3H),7.05(t,J=7.7Hz,1H),6.48(d,J=7.4Hz,1H),6.40–6.31(m,2H), 4.69(p,J=7.3Hz,1H),4.44–4.36(m,4H),2.21(s,3H),2.19(s,6H).LRMS(ESI+ )m / z:461.0[M+1] + .

[0220] Example 27: N-(2,6-dimethylphenyl)-1-(1-(p-methylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P27)

[0221]

[0222] The synthesis method follows general procedure C, using 4-methyl-1-bromobenzene as the corresponding aryl bromide. White solid (88%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.92(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.69(d,J=8.3Hz,1H),7.64–7.58(m,1H),7.31(t,J=7.5 Hz,1H),7.13(s,3H),6.98(d,J=8.0Hz,2H),6.50–6.45(m,2H),4.68(p,J=7.1Hz,1H),4.37(d,J=7.2Hz,4H),2.18(s,6H),2.15(s,3H).LRMS(ESI + )m / z:461.0[M+1] + .

[0223] Example 28: N-(2,6-dimethylphenyl)-1-(1-(4-ethylphenyl)azacyclobutane-3-substituted)-9H-pyridine[3,4-b]indole-3-amide (P28)

[0224]

[0225] The synthesis method follows general procedure C, using 4-ethyl-1-bromobenzene as the corresponding aryl bromide. White solid (84%). 1H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.94(s,1H),8.83(s,1H),8.41(d,J=7.9H z,1H),7.69(d,J=8.2Hz,1H),7.61(t,J=7.6Hz,1H),7.31(t,J=7.5Hz,1H),7.12( s,3H),7.01(d,J=7.9Hz,2H),6.49(d,J=8.0Hz,2H),4.75–4.62(m,1H),4.37(d,J =7.2Hz,4H),2.45(q,J=7.6Hz,2H),2.17(s,6H),1.10(t,J=7.6Hz,3H).LRMS(ESI + m / z:475.0[M+1] + .

[0226] Example 29: N-(2,6-dimethylphenyl)-1-(1-(2-ethylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P29)

[0227]

[0228] The synthesis method follows general procedure C, using 2-ethyl-1-bromobenzene as the corresponding aryl bromide. White solid (74%). 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),10.02(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.69(d ,J=8.2Hz,1H),7.62(ddd,J=8.3,7.0,1.2Hz,1H),7.35–7.28(m,1H),7.13(s,3H),7.05(td,J=7. 7,6.0Hz,2H),6.76–6.70(m,1H),6.65(d,J=8.0Hz,1H),4.66(t,J=7.5Hz,1H),4.48(t,J=6.9Hz ,2H),4.44(t,J=7.4Hz,2H),2.61(q,J=7.5Hz,2H),2.20(s,6H),1.19(t,J=7.4Hz,3H).LRMS(ESI + m / z:475.0[M+1] + .

[0229] Example 30: N-(2,6-dimethylphenyl)-1-(1-(3-ethylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P30)

[0230]

[0231] The synthesis method follows general procedure C, using 3-ethyl-1-bromobenzene as the corresponding aryl bromide. White solid (83%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.98(s,1H),8.84(s,1H),8.41(d,J=7.9Hz,1H), 7.69(d,J=8.2Hz,1H),7.62(ddd,J=8.1,7.0,1.2Hz,1H),7.32(t,J=7.5Hz,1H),7.13(s, 3H),7.07(t,J=7.7Hz,1H),6.52(d,J=7.5Hz,1H),6.41–6.34(m,2H),4.69(p,J=7.1Hz, 1H),4.44–4.37(m,4H),2.55–2.48(m,2H),2.19(s,6H),1.14(t,J=7.6Hz,3H).LRMS(ESI + m / z:475.0[M+1] + .

[0232] Example 31: N-(2,6-dimethylphenyl)-1-(1-(2-isopropylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P31)

[0233]

[0234] The synthesis method follows general procedure C, using 2-isopropyl-1-bromobenzene as the corresponding aryl bromide. White solid (50%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),10.03(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.69 (d,J=8.2Hz,1H),7.65–7.59(m,1H),7.32(t,J=7.5Hz,1H),7.18–7.12(m,4H),7.05(t,J=7.5H z,1H),6.79(t,J=7.5Hz,1H),6.71(d,J=8.1Hz,1H),4.63(p,J=7.3Hz,1H),4.48(t,J=6.7Hz,2 H),4.43(t,J=7.4Hz,2H),3.18(p,J=6.9Hz,1H),2.21(s,6H),1.20(d,J=6.7Hz,6H).LRMS(ESI+ m / z:489.0[M+1] + .

[0235] Example 32: N-(2,6-dimethylphenyl)-1-(1-(3-isopropylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P32)

[0236]

[0237] The synthesis method follows general procedure C, using 3-isopropyl-1-bromobenzene as the corresponding aryl bromide. White solid (62%). 1 H NMR (400MHz, DMSO-d6) δ11.98(s,1H),9.99(s,1H),8.84(s,1H),8.41(d,J=7.9Hz,1H),7.69(d ,J=8.3Hz,1H),7.62(ddd,J=8.2,7.0,1.3Hz,1H),7.32(t,J=7.4Hz,1H),7.12(s,3H),7.08(t, J=7.7Hz,1H),6.56(d,J=7.5Hz,1H),6.41(d,J=2.1Hz,1H),6.37(dd,J=7.7,2.2Hz,1H),4.69( p,J=7.2Hz,1H),4.46–4.31(m,4H),2.78(p,J=7.0Hz,1H),2.18(s,6H),1.16(d,J=6.9Hz,6H). 13 C NMR(151MHz,DMSO-d6)δ163.26,152.30,149.08,143.33,141.03,138.72,135.49,135.25,135.20,128.68,128.62,128.29,12 7.74,126.41,122.33,121.46,120.21,115.19,113.16,112.34,109.45,109.20,56.08,33.73,32.19,23.96,18.41.LRMS(ESI + m / z:489.0[M+1] + .

[0238] Example 33: N-(2,6-dimethylphenyl)-1-(1-(4-isopropylphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P33)

[0239]

[0240] The synthesis method follows general procedure C, using 4-isopropyl-1-bromobenzene as the corresponding aryl bromide. White solid (66%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.95(s,1H),8.83(s,1H),8.41(d,J=8.0H z,1H),7.69(d,J=8.2Hz,1H),7.61(t,J=7.6Hz,1H),7.31(t,J=7.4Hz,1H),7.12 (s,3H),7.07–7.01(m,2H),6.50(d,J=7.7Hz,2H),4.69(p,J=7.2Hz,1H),4.37(d ,J=7.2Hz,4H),2.82–2.68(m,1H),2.17(s,6H),1.13(d,J=6.9Hz,6H).LRMS(ESI + m / z:489.0[M+1] + .

[0241] Example 34: N-(2,6-dimethylphenyl)-1-(1-(4-methoxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P34)

[0242]

[0243] The synthesis method follows general procedure C, using 4-methoxy-1-bromobenzene as the corresponding aryl bromide. White solid (69%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.96(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.61(ddd,J=8.2,7.1,1.2Hz,1H),7.32 (t,J=7.5Hz,1H),7.13(s,3H),6.87–6.75(m,2H),6.57–6.47(m,2H),4.66( t,J=7.2Hz,1H),4.34(d,J=7.2Hz,4H),3.64(s,3H),2.19(s,6H).LRMS(ESI + )m / z:477.0[M+1] + .

[0244] Example 35: N-(2,6-dimethylphenyl)-1-(1-(2-methoxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P35)

[0245]

[0246] The synthesis method follows general procedure C, using 2-methoxy-1-bromobenzene as the corresponding aryl bromide. White solid (49%). 1 H NMR(400MHz,DMSO-d6)δ11.97(s,1H),10.01(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7 .69(d,J=8.2Hz,1H),7.61(ddd,J=8.2,7.0,1.2Hz,1H),7.34–7.29(m,1H),7.13(s,3H),6. 85(dd,J=8.1,1.5Hz,1H),6.81(dd,J=7.5,1.4Hz,1H),6.76–6.68(m,1H),6.58(dd,J=7.8 ,1.7Hz,1H),4.62(p,J=7.4Hz,1H),4.48–4.37(m,4H),3.73(s,3H),2.20(s,6H).LRMS(ESI + m / z: 476.9 [M+1] + .

[0247] Example 36: N-(2,6-dimethylphenyl)-1-(1-(3-methoxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P36)

[0248]

[0249] The synthesis method follows general procedure C, using 3-methoxy-1-bromobenzene as the corresponding aryl bromide. White solid (63%). 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),9.98(s,1H),8.84(s,1H),8.41(d,J=7.9Hz ,1H),7.69(d,J=8.2Hz,1H),7.62(t,J=7.6Hz,1H),7.32(t,J=7.4Hz,1H),7.13(s, 3H),7.06(t,J=8.1Hz,1H),6.24(d,J=8.2Hz,1H),6.14(d,J=8.0Hz,1H),6.08(s, 1H),4.73–4.64(m,1H),4.41(p,J=7.0Hz,4H),3.69(s,3H),2.19(s,6H).LRMS(ESI + m / z: 476.9 [M+1]+ .

[0250] Example 37: 1-(1-(2-(benzyloxy)phenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P37)

[0251]

[0252] The synthesis method follows general procedure C, using 2-benzyloxy-1-bromobenzene as the corresponding aryl bromide. White solid (50%). 1 H NMR (600MHz, DMSO-d6) δ12.00(s,1H),9.97(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.68(d,J=8.2H z,1H),7.61(ddd,J=8.3,7.0,1.2Hz,1H),7.47–7.43(m,2H),7.30(q,J=7.3Hz,3H),7.25–7.21(m,1H),7 .13(s,3H),6.94(dd,J=8.1,1.3Hz,1H),6.83(td,J=7.6,1.3Hz,1H),6.70(td,J=7.7,1.6Hz,1H),6.61( dd,J=7.8,1.6Hz,1H),5.05(s,2H),4.63(p,J=7.5Hz,1H),4.44(p,J=7.4Hz,4H),2.19(s,6H).LRMS(ESI + m / z:552.8[M+1] + .

[0253] Example 38: N-(2,6-methylphenyl)-1-(1-(2-hydroxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P38)

[0254]

[0255] Compound P37 (20 mg) was dissolved in methanol, and 10 mg of 10% Pd / C was added. The mixture was hydrogenated overnight at room temperature. The reaction was monitored by TLC. Palladium on carbon and insoluble matter were filtered off. The reaction solution was concentrated and purified by silica gel column chromatography to obtain the white target product. White solid (68%). 1H NMR(600MHz,DMSO-d6)δ11.97(s,1H),10.03(s,1H),8.95(s,1H),8.83(s,1H ),8.41(d,J=7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.61(ddd,J=8.3,7.0,1.2Hz, 1H),7.37–7.28(m,1H),7.13(s,3H),6.67(t,J=7.6Hz,2H),6.61–6.52(m,2H ),4.59(p,J=7.4Hz,1H),4.42(dt,J=24.2,7.3Hz,4H),2.21(s,6H).LRMS(ESI + m / z:462.8[M+1] + .

[0256] Example 39: 1-(1-(2-cyanophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P39)

[0257]

[0258] The synthesis method follows general procedure C, using 2-cyano-1-bromobenzene as the corresponding aryl bromide. White solid (70%). 1 H NMR (400MHz, DMSO-d6) δ12.04(s,1H),10.05(s,1H),8.85(s,1H),8.42(d,J=7.9Hz,1H),7.70(d,J=8.2Hz,1H),7.62(ddd,J=8.2,7.0,1.2Hz,1H) ,7.50(dd,J=7.9,1.5Hz,1H),7.48–7.41(m,1H),7.32(t,J=7.3Hz,1H),7 .13(s,3H),6.81–6.72(m,2H),4.79–4.62(m,5H),2.21(s,6H).LRMS(ESI + m / z:472.0[M+1] + .

[0259] Example 40: 1-(1-(4-cyanophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P40)

[0260]

[0261] The synthesis method follows general procedure C, using 4-cyano-1-bromobenzene as the corresponding aryl bromide. White solid (81%). 1 H NMR (400MHz, DMSO-d6) δ12.04(s,1H),10.02(s,1H),8.85(s,1H),8.42(d,J=7 .9Hz,1H),7.69(d,J=8.2Hz,1H),7.65–7.59(m,1H),7.55(d,J=8.7Hz,2H),7.3 2(t,J=7.5Hz,1H),7.13(s,3H),6.58(d,J=8.8Hz,2H),4.75(p,J=7.9,7.2Hz, 1H),4.62(dd,J=7.7,5.9Hz,2H),4.51(t,J=8.1Hz,2H),2.19(s,6H).LRMS(ESI + m / z:472.0[M+1] + .

[0262] Example 41: N-(2,6-dimethylphenyl)-1-(1-(2-(trifluoromethyl)phenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P41)

[0263]

[0264] The synthesis method follows general procedure C, using 2-trifluoromethyl-1-bromobenzene as the corresponding aryl bromide. White solid (58%). 1 H NMR(400MHz,DMSO-d6)δ12.03(s,1H),10.04(s,1H),8.85(s,1H),8.42(d,J =7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.65–7.59(m,1H),7.50(dd,J=8.2,1.6 Hz,1H),7.46(t,J=7.7Hz,1H),7.32(t,J=7.5Hz,1H),7.13(s,3H),6.84–6. 79(m,2H),4.73–4.62(m,3H),4.57(d,J=6.9Hz,2H),2.21(s,6H).LRMS(ESI + m / z: 514.9 [M+1] + .

[0265] Example 42: N-(2,6-dimethylphenyl)-1-(1-(2-(methanesulfonyl)phenyl)azacyclobutane-3-yl)-9H-pyrido[3,4-b]indole-3-amide (P42)

[0266]

[0267] The synthesis method follows general procedure C, using 2-methanesulfonyl-1-bromobenzene as the corresponding aryl bromide. White solid (60%). 1 H NMR (400MHz, DMSO-d6) δ12.05(s,1H),10.08(s,1H),8.85(s,1H),8.42(d,J=7. 9Hz,1H),7.78(d,J=8.0Hz,1H),7.69(d,J=8.2Hz,1H),7.62(t,J=7.6Hz,1H),7 .53(t,J=7.7Hz,1H),7.32(t,J=7.5Hz,1H),7.14(s,3H),7.03(d,J=8.4Hz,1H) ,6.93(t,J=7.6Hz,1H),4.77–4.61(m,5H),3.31(s,3H),2.21(s,6H).LRMS(ESI + m / z:524.9[M+1] + .

[0268] Example 43: N-(2,6-dimethylphenyl)-1-(1-(4-(methylsulfonyl)phenyl)azacyclobutane-3-yl)-9H-pyrido[3,4-b]indole-3-amide (P43)

[0269]

[0270] The synthesis method follows general procedure C, using 4-methanesulfonyl-1-bromobenzene as the corresponding aryl bromide. White solid (72%). 1 H NMR (400MHz, DMSO-d6) δ12.04(s,1H),10.03(s,1H),8.86(s,1H),8.42(d,J=7.9Hz,1H),7.72–7.58(m,4H),7.36–7.28(m,1H),7.13( s,3H),6.68–6.61(m,2H),4.82–4.71(m,1H),4.63(dd,J=7.7,6.0Hz,2H),4.52(t,J=8.0Hz,2H),3.05(s,3H),2.19(s,6H).LRMS(ESI + m / z:524.9[M+1] + .

[0271] Example 44: N-(2,6-dimethylphenyl)-1-(1-(4-(trifluoromethyl)phenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P44)

[0272]

[0273] The synthesis method follows general procedure C, using 4-trifluoromethyl-1-bromobenzene as the corresponding aryl bromide. White solid (70%). 1 H NMR(600MHz,DMSO-d6)δ12.03(s,1H),10.00(s,1H),8.86(s,1H),8.42(d,J=7.8Hz,1H ),7.69(dt,J=8.3,0.9Hz,1H),7.62(ddd,J=8.2,7.0,1.2Hz,1H),7.48(d,J=8.5Hz,2H) ,7.32(ddd,J=8.0,7.0,1.0Hz,1H),7.12(s,3H),6.64(d,J=8.5Hz,2H),4.75(tt,J=8.3 ,6.1Hz,1H),4.57(dd,J=7.4,6.1Hz,2H),4.49(t,J=7.9Hz,2H),2.18(s,6H).LRMS(ESI + m / z: 515.2 [M+1] + .

[0274] Example 45: 1-(1-(4-(dimethylamino)phenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P45)

[0275]

[0276] The synthesis method follows general procedure C, using 4-dimethylamino-1-bromobenzene as the corresponding aryl bromide. White solid (34%). 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),9.94(s,1H),8.82(s,1H),8.41(d,J=7.9Hz,1H),7.69(d,J=8.2Hz,1H),7.61(t,J=7.6Hz,1H),7.31(t ,J=7.5Hz,1H),7.13(s,3H),6.74–6.62(m,2H),6.58–6.43(m,2H),4.74–4.57(m,1H),4.40–4.23(m,4H),2.73(s,6H),2.18(s,6H).LRMS(ESI+ m / z:489.8[M+1] + .

[0277] Example 46: N-(2,6-dimethylphenyl)-1-(1-(4-fluoro-3-methoxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P46)

[0278]

[0279] The synthesis method follows general procedure C, using 3-methoxy-4-fluoro-1-bromobenzene as the corresponding aryl bromide. White solid (21%). 1 H NMR (400MHz, DMSO-d6) δ12.00(s,1H),9.99(s,1H),8.84(s,1H),8.41(d,J=7.9Hz, 1H),7.69(d,J=8.3Hz,1H),7.62(t,J=7.7Hz,1H),7.32(t,J=7.5Hz,1H),7.13(s,3 H),6.99(dd,J=11.5,8.7Hz,1H),6.28(dd,J=7.5,2.5Hz,1H),6.04(dd,J=8.0,4.6 Hz,1H),4.75–4.58(m,1H),4.45–4.25(m,4H),3.79(s,3H),2.19(s,6H).LRMS(ESI + m / z:494.9[M+1] + .

[0280] Example 47: N-(2,6-dimethylphenyl)-1-(1-(4-fluoro-2-methoxyphenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P47)

[0281]

[0282] The synthesis method follows general procedure C, using 2-methoxy-4-fluoro-1-bromobenzene as the corresponding aryl bromide. White solid (17%). 1H NMR (400MHz, DMSO-d6) δ11.97(s,1H),10.01(s,1H),8.82(s,1H),8.41(d,J=7.9Hz, 1H),7.69(d,J=8.2Hz,1H),7.61(t,J=7.7Hz,1H),7.31(t,J=7.5Hz,1H),7.13(s,3H ),6.79(dd,J=10.7,2.7Hz,1H),6.64(td,J=8.5,2.8Hz,1H),6.55(dd,J=8.7,6.0Hz ,1H),4.61(p,J=7.6Hz,1H),4.45–4.29(m,4H),3.75(s,3H),2.21(s,6H).LRMS(ESI + m / z:494.9[M+1] + .

[0283] Example 48: 1-(1-(2,6-difluorophenyl)azacyclobutane-3-substituted)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P48)

[0284]

[0285] The synthesis method follows general procedure C, using 2,6-difluoro-1-bromobenzene as the corresponding aryl bromide. White solid (12%). 1 H NMR (400MHz, DMSO-d6) δ11.97(s,1H),10.13(s,1H),8.83(s,1H),8.41(d,J=7.9Hz,1H),7.68(d,J=8.2Hz,1H),7.61(t,J=7.7Hz,1H ),7.31(t,J=7.5Hz,1H),7.13(s,3H),6.93(td,J=8.4,2.5Hz,2H),6.69(q,J=7.0Hz,1H),4.82–4.63(m,5H),2.22(s,6H).LRMS(ESI + m / z: 482.9 [M+1] + .

[0286] Example 49: N-(2,6-dimethylphenyl)-1-(1-(4-(S-aminomethylsulfonyl)phenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P49)

[0287]

[0288] The synthesis of P49-1 follows the general procedure C, using 4-(N-Boc-S-methylsulfonylimideyl)bromobenzene as the corresponding aryl bromide. P49-1 was then dissolved in dichloromethane, and trifluoroacetic acid (5:1 v / v) was added under ice bath conditions. The reaction was carried out at room temperature, and the reaction was monitored by TLC. After completion, saturated sodium carbonate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to obtain P49. 1 H NMR(600MHz,DMSO-d6)δ12.05(s,1H),10.03(s,1H),8.86(s,1H),8.42(d,J=7.9H z,1H),7.72–7.68(m,3H),7.62(ddd,J=8.3,7.0,1.2Hz,1H),7.34–7.30(m,1H),7. 13(s,3H),6.64–6.60(m,2H),4.76(tt,J=8.3,6.1Hz,1H),4.60(td,J=7.0,6.4,3 .5Hz,2H),4.50(t,J=8.0Hz,2H),3.83(s,1H),2.94(s,3H),2.19(s,6H).LRMS(ESI + m / z:524.3[M+1] + .

[0289] Example 50: N-(2,6-dimethylphenyl)-1-(1-(4-amidophenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P50)

[0290]

[0291] The synthesis method for P50-1 follows the general procedure C, using 4-(N,N-di-p-methoxybenzylaminosulfonyl)bromobenzene as the corresponding aryl bromide. P50-1 was dissolved in dichloromethane, and trifluoroacetic acid (5:1 v / v) was added under ice bath conditions. The reaction was carried out at room temperature, and the reaction was monitored by TLC. After the reaction was complete, saturated sodium carbonate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to obtain P50. 1H NMR (600MHz, DMSO-d6) δ12.04(s,1H),10.05(s,1H),8.86(s,1H),8.42(d,J= 7.9Hz,1H),7.70(d,J=8.2Hz,1H),7.62(dd,J=8.1,5.9Hz,3H),7.32(t,J=7.5 Hz,1H),7.13(s,3H),7.01(s,2H),6.60(d,J=8.7Hz,2H),4.74(td,J=8.3,4. 2Hz,1H),4.58(t,J=6.8Hz,2H),4.48(t,J=7.9Hz,2H),2.20(s,6H).LRMS(ESI + m / z:525.8[M+1] + .

[0292] Example 51: N-(2,6-dimethylphenyl)-1-(1-(4-(piperidin-4-substituted)phenyl)azacyclobutane-3-yl)-9H-pyrido[3,4-b]indole-3-amide (P51)

[0293]

[0294] The synthesis of P51-1 follows the general procedure D, using 4-(N-Boc-piperidin-4-yl)bromobenzene as the corresponding aryl bromide. P51-1 was dissolved in 5 mL of dichloromethane, and 1 mL of trifluoroacetic acid was added under ice bath conditions. The reaction was carried out at room temperature, and the reaction was monitored by TLC. After the reaction was complete, saturated sodium carbonate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to obtain P51. 1 H NMR(600MHz,DMSO-d6)δ12.04(s,1H),9.96(s,1H),8.84(s,1H),8.41(s, 1H),7.78–7.55(m,2H),7.31(s,1H),7.12(s,3H),7.02(s,2H),6.50(s,2H ),4.69(s,1H),4.38(s,4H),3.08–2.87(m,2H),2.62–2.51(m,2H),2.46– 2.39(m,1H),2.17(s,6H),1.70–1.53(m,2H),1.49–1.36(m,2H).LRMS(ESI + m / z:529.9[M+1] + .

[0295] Example 52: N-(2,6-dimethylphenyl)-1-(1-(4-(piperazin-1-carbonyl)phenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P52)

[0296]

[0297] The synthesis of P52-1 follows the general procedure C, using 4-(4-Boc-piperazin-1-ylcarbonyl)bromobenzene as the corresponding aryl bromide. P52-1 was dissolved in dichloromethane, and trifluoroacetic acid (5:1 v / v) was added under ice bath conditions. The reaction was carried out at room temperature, and the reaction was monitored by TLC. After the reaction was complete, saturated sodium carbonate was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography. 1 H NMR (600MHz, DMSO-d6) δ12.04(s,1H),10.02(s,1H),8.85(s,1H),8.42(d,J=7.8Hz ,1H),7.69(d,J=8.2Hz,1H),7.62(t,J=7.7Hz,1H),7.32(t,J=7.5Hz,1H),7.25(d,J =8.1Hz,2H),7.12(s,3H),6.55(d,J=8.1Hz,2H),4.74(t,J=7.6Hz,1H),4.51(t,J= 6.8Hz,2H),4.45(t,J=7.9Hz,2H),3.39(s,4H),2.66(s,4H),2.18(s,6H).LRMS(ESI + m / z:559.3[M+1] + .

[0298] Example 53: N-(2,6-dimethylphenyl)-1-(1-(4-(4-methylpiperazinyl-1-carbonyl)phenyl)azacyclobutane-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P53)

[0299]

[0300] The synthesis method for P53-1 is the same as that for general procedure C, using 4-carboxybromobenzene as the corresponding aryl bromide. The synthesis method for P53 is the same as that for intermediate I1, using 1-methylpiperazine instead of 2,6-dimethylaniline. 1H NMR(600MHz,DMSO-d6)δ12.09(s,1H),10.02(s,1H),8.85(s,1H),8.42(d,J=7.9Hz,1H), 7.70(d,J=8.2Hz,1H),7.62(t,J=7.7Hz,1H),7.32(t,J=7.6Hz,1H),7.27(d,J=8.2Hz,2H ),7.13(d,J=3.4Hz,3H),6.55(d,J=8.1Hz,2H),4.74(q,J=7.3Hz,1H),4.52(t,J=6.8Hz, 2H),4.46(t,J=7.8Hz,2H),3.52(s,4H),2.45(s,4H),2.29(s,3H),2.18(s,6H).LRMS(ESI + m / z: 572.8 [M+1] + .

[0301] Example 54: N-(2,6-dimethylphenyl)-1-(1-(4-(morpholino-4-carbonyl)phenyl)azacyclobutane-3-yl)-9H-pyrido[3,4-b]indole-3-amide (P54)

[0302]

[0303] The synthesis of compound P53 was referenced in the preparation of compound P54 by replacing 2,6-dimethylaniline with morpholine after preparing P53-1, as described in reference I1. 1 H NMR (600MHz, DMSO-d6) δ12.04(s,1H),10.02(s,1H),8.85(s,1H),8.42(d,J=7.8Hz,1H) ,7.69(d,J=8.2Hz,1H),7.62(ddd,J=8.2,6.9,1.2Hz,1H),7.34–7.30(m,1H),7.30–7.27 (m,2H),7.13(s,3H),6.57–6.54(m,2H),4.74(tt,J=8.2,6.2Hz,1H),4.52(t,J=6.8Hz, 2H),4.46(t,J=7.8Hz,2H),3.59–3.55(m,4H),3.50–3.44(m,4H),2.18(s,6H).LRMS(ESI + m / z:560.0[M+1] + .

[0304] Example 55: Synthesis of compound P55:

[0305]

[0306] Dissolve a6 (1.5 g, 3.69 mmol) in 15 mL of dichloromethane, add 3 mL of trifluoroacetic acid under ice bath, stir overnight at room temperature, and monitor the reaction by TLC. After the reaction is complete, concentrate the reaction solution to obtain crude trifluoroacetate. Dissolve the crude product in acetonitrile, add 3-phenylpropanal (4.43 mmol) as the substrate aldehyde, and react overnight. After the reaction is complete by TLC, concentrate the reaction solution by rotary evaporation, quench the reaction with saturated sodium carbonate solution, adjust the pH to 10-12, extract four times with dichloromethane (50 mL × 4), combine the organic phases, dry with anhydrous sodium sulfate, concentrate the organic phase to obtain an oily liquid crude product, dissolve the crude product in DMF, add triethylamine (1.2 mL, 9.23 mmol), add NCS (1 g, 7.75 mmol) under ice bath, remove the ice bath, and react at room temperature for 1 to 2 hours, monitoring the reaction by TLC. After the reaction was complete, 60 mL of saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (20 mL × 3). The organic phases were combined, washed three times with saturated brine (40 mL × 3), and dried with anhydrous sodium sulfate. The organic phase was concentrated to obtain an oily crude liquid, which was then purified by silica gel column chromatography to obtain the white target product P55 (50%).

[0307] Example 56: N-(2,6-dimethylphenyl)-1-(2-(thiazol-5-substituted)ethyl)-9H-pyrido[3,4-b]indole-3-amide (P56)

[0308]

[0309] Synthesized as in compound P55, using 5-propionaldehyde thiazole as the substrate aldehyde. White solid (66%). 1 H NMR (400MHz, DMSO-d6) δ12.02(s,1H),10.10(s,1H),8.83(s,1H),8.80(s,1H),8.38(d,J=7.9Hz,1H),7.76(s,1H),7.6 8–7.55(m,2H),7.30(t,J=7.6Hz,1H),7.15(s,3H),3.74(t,J=7.3Hz,2H),3.57(t,J=7.3Hz,2H),2.24(s,6H).LRMS(ESI + m / z: 426.8 [M+1] + .

[0310] Example 57: N-(2,6-dimethylphenyl)-1-(2-(thiazol-2-substituted)ethyl)-9H-pyrido[3,4-b]indole-3-amide (P57)

[0311]

[0312] Synthesized as in compound P55, using 2-propanalthiazole as the substrate aldehyde. White solid (68%). 1 H NMR (400MHz, DMSO-d6) δ12.08(s,1H),10.06(s,1H),8.78(s,1H),8.38(d,J=7.9Hz,1H),7.69–7.63(m,2H),7.59(t,J=7.7Hz,1H ),7.51(d,J=3.2Hz,1H),7.30(t,J=7.5Hz,1H),7.15(s,3H),3.89(t,J=7.2Hz,2H),3.71(t,J=7.4Hz,2H),2.23(s,6H).LRMS(ESI + m / z: 426.8 [M+1] + .

[0313] Example 58: 1-(2-(1H-imidazol-5-substituted)ethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P58)

[0314]

[0315] The synthesis method is the same as that for compound P55, using 5-propionaldehyde imidazole as the substrate aldehyde. White solid (66%). 1 H NMR (400MHz, DMSO-d6) δ12.08(s,1H),10.06(s,1H),8.77(s,1H),8.37(d,J=7.8Hz,1H),7.66(d,J=8.0Hz,1H),7.62–7. 52(m,2H),7.29(t,J=7.3Hz,1H),7.14(s,3H),6.83(s,1H),3.57–3.52(m,2H),3.30–3.25(m,2H),2.24(s,6H).LRMS(ESI + m / z:409.8[M+1] + .

[0316] Example 59: 1-(2-(1H-imidazol-2-substituted)ethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P59)

[0317]

[0318] The synthetic method was similar to that of compound P55, using 2-propionaldehyde imidazole as the substrate aldehyde. A white target product was obtained.1 HNMR (400MHz, DMSO-d6) δ12.08(s,1H),11.80(s,1H),10.01(s,1H),8.75(s,1H),8.37(d,J=7.9Hz,1H),7.66(dt,J=8.2,1.0Hz,1H),7.59(ddd,J=8. 2,6.9,1.2Hz,1H),7.29(ddd,J=8.0,7.0,1.1Hz,1H),7.15(s,3H),6.84(s ,2H),3.65(t,J=7.4Hz,2H),3.40(t,J=7.4Hz,2H),2.23(s,6H).LRMS(ESI + m / z:409.8[M+1] + .

[0319] Example 60: N-cyclopropyl-1-phenylethyl-9H-pyrido[3,4-b]indole-3-amide (P60)

[0320]

[0321] The synthesis method is based on intermediate a6 (step i of preparation method two) and compound P55, using cyclopropylamine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(600MHz,Chloroform-d)δ8.95(s,1H),8.76(s,1H),8.18(d,J=3.7Hz,1H),8.04(d, J=7.9Hz,1H),7.50–7.47(m,1H),7.45(d,J=8.1Hz,1H),7.24(d,J=8.4Hz,1H),7.19(d, J=6.0Hz,4H),7.14(dq,J=6.0,2.7Hz,1H),3.45(t,J=7.6Hz,2H),3.22(t,J=7.6Hz,2H) ,2.93(tq,J=7.4,3.8Hz,1H),0.81(td,J=7.0,5.1Hz,2H),0.68–0.64(m,2H).LRMS(ESI + m / z: 356.8 [M+1] + .

[0322] Example 61: N-cyclohexyl-1-phenylethyl-9H-pyrido[3,4-b]indole-3-amide (P61)

[0323]

[0324] The synthesis method is based on intermediate a6 and compound P55, using cyclohexylamine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 HNMR(600MHz,Chloroform-d)δ8.75(s,1H),8.52(s,1H),8.11(d,J=8.6Hz,1H),8.05(d,J=7.9Hz, 1H),7.48(t,J=7.6Hz,1H),7.41(d,J=8.2Hz,1H),7.25–7.23(m,1H),7.20(p,J=7.0,6.3Hz,4H),7. 17–7.11(m,1H),4.02(qd,J=9.7,4.3Hz,1H),3.44(t,J=7.5Hz,2H),3.24(t,J=7.5Hz,2H),2.05–1. 99(m,2H),1.74(dt,J=13.2,3.7Hz,2H),1.62(dt,J=13.3,3.9Hz,1H),1.44–1.24(m,6H).LRMS(ESI + m / z:397.7[M+1] + .

[0325] Example 62: 1-Phenylacetyl-N-(piperidin-4-substituted)-9H-pyrido[3,4-b]indole-3-amide (P62)

[0326]

[0327] The synthesis method is based on intermediate a6 and compound P55, using 4-aminopiperidine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, Methanol-d4) δ9.31 (s, 1H), 8.44 (d, J = 7.9Hz, 1H), 7.85 (t, J = 7.3H z,1H),7.80(d,J=8.5Hz,1H),7.54(t,J=7.5Hz,1H),7.22(s,3H),7.15(s,1H),4. 32(d,J=12.6Hz,1H),3.84(d,J=8.2Hz,2H),3.56(d,J=12.6Hz,2H),3.24(q,J=1 3.0,10.5Hz,4H),2.31(d,J=13.8Hz,2H),2.05(q,J=12.2,11.6Hz,2H).LRMS(ESI + m / z:398.9[M+1] + .

[0328] Example 63: Morpholine (1-phenylethyl-9H-pyridine[3,4-b]indole-3-substituted) methyl ketone (P63)

[0329]

[0330] The synthesis method is based on intermediate a6 and compound P55, using morpholine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(500MHz,Chloroform-d)δ8.51(s,1H),8.26(s,1H),7.99(d,J=7.9Hz,1H),7.54–7.46(m,1H),7.41(d,J=8.2Hz,1H),7.26(t,J=3. 8Hz,1H),7.24–7.19(m,2H),7.18–7.11(m,3H),3.89–3.77(m,4H),3.65(s,4H),3.40(t,J=7.5Hz,2H),3.20(t,J=7.5Hz,2H).LRMS(ESI + m / z: 385.9 [M+1] + .

[0331] Example 64: 1-Phenylacetyl-N-(tetrahydro-2H-pyran-4-substituted)-9H-pyrido[3,4-b]indole-3-amide (P64)

[0332]

[0333] The synthesis method is based on intermediate a6 and compound P55, using 4-aminotetrahydropyran condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(600MHz,Chloroform-d)δ8.77(s,1H),8.38(s,1H),8.11(t,J=7.1Hz,2H),7.54–7.50(m,1 H),7.42(d,J=8.2Hz,1H),7.29(t,J=7.5Hz,1H),7.26–7.24(m,2H),7.22–7.18(m,3H),4.26(d ddd,J=15.2,10.8,8.4,4.3Hz,1H),4.02(dt,J=11.9,3.5Hz,2H),3.56(td,J=11.6,2.2Hz,2H) ,3.46(t,J=7.5Hz,2H),3.26(t,J=7.5Hz,2H),2.06–2.00(m,2H),1.69–1.65(m,2H).LRMS(ESI + m / z:399.8[M+1] + .

[0334] Example 65: 1-Phenylacetyl-N-(pyridine-4-substituted)-9H-pyrido[3,4-b]indole-3-amide (P65)

[0335]

[0336] The synthesis method is based on intermediate a6 and compound P55, using 4-aminopyridine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.14(s,1H),10.65(s,1H),8.87(s,1H),8.56–8.49(m,2H),8.40(d,J=7.9Hz,1H),7.98–7.91(m,2H),7.67(d,J=8.2Hz ,1H),7.63–7.59(m,1H),7.45–7.40(m,2H),7.31(td,J=7.6,2.1Hz,3H),7.19(t,J=7.3Hz,1H),3.59–3.54(m,2H),3.34–3.30(m,2H).LRMS(ESI + m / z:392.9[M+1] + .

[0337] Example 66: 1-Phenylacetyl-N-(pyridine-2-substituted)-9H-pyrido[3,4-b]indole-3-amide (P66)

[0338]

[0339] The synthesis method is based on intermediate a6 and compound P55, using 2-aminopyridine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(600MHz,DMSO-d6)δ12.14(s,1H),10.64(s,1H),8.89(s,1H),8.43–8.40(m, 2H),8.39–8.36(m,1H),7.91(ddd,J=8.7,7.4,1.9Hz,1H),7.66(d,J=8.2Hz,1H), 7.61(ddd,J=8.2,6.9,1.2Hz,1H),7.43–7.39(m,2H),7.31(q,J=8.0Hz,3H),7.21 –7.16(m,2H),3.55(dd,J=9.5,6.7Hz,2H),3.29(dd,J=9.5,6.7Hz,2H).LRMS(ESI + m / z:392.9[M+1] + .

[0340] Example 67: 1-Phenylacetyl-N-(pyridine-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P67)

[0341]

[0342] The synthesis method is based on intermediate a6 and compound P55, using 3-aminopyridine condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.11(s,1H),10.56(s,1H),9.08(d,J=2.4Hz,1H),8.86(s,1H),8.40(d,J=7.9Hz,1H),8.37–8.31(m,2H),7.66(d,J=8.2Hz ,1H),7.60(t,J=7.6Hz,1H),7.47–7.39(m,3H),7.37–7.29(m,3H),7.19( t,J=7.4Hz,1H),3.56(dd,J=9.9,6.4Hz,2H),3.35–3.31(m,2H).LRMS(ESI + m / z:392.9[M+1] + .

[0343] Example 68: N-(1H-imidazol-2-substituted)-1-phenethyl-9H-pyrido[3,4-b]indole-3-amide (P68)

[0344]

[0345] The synthesis method is based on intermediate a6 and compound P55, using 2-aminoimidazolium condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.14(s,1H),8.85(s,1H),8.40(d,J=7.9Hz,1H),7.67(d,J=8.2Hz,1H),7.61(ddd,J=8.2,6.9,1.2Hz,1H),7. 44–7.38(m,2H),7.31(td,J=7.4,2.2Hz,3H),7.22–7.16(m,1H),6.86(s,2H),3.56–3.50(m,2H),3.28(dd,J=9.5,6.8Hz,2H).LRMS(ESI + m / z:381.9[M+1] + .

[0346] Example 69: 1-Phenylacetyl-N-(thiazol-2-substituted)-9H-pyrido[3,4-b]indole-3-amide (P69)

[0347]

[0348] The synthesis method is based on intermediate a6 and compound P55, using 2-aminothiazole condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (500MHz, DMSO-d6) δ12.17(s,1H),11.64(s,1H),8.88(s,1H),8.40(d,J=7.8Hz,1H),7.67(d,J=8.1Hz,1H),7.64–7.56 (m,2H),7.44–7.39(m,2H),7.36–7.27(m,4H),7.22–7.15(m,1H),3.57–3.50(m,2H),3.30(dd,J=9.7,6.5Hz,2H).LRMS(ESI + )m / z:398.8[M+1] + .

[0349] Example 70: N-(1-methyl-1H-pyrazole-4-substituted)-1-phenethyl-9H-pyrido[3,4-b]indole-3-amide (P70)

[0350]

[0351] The synthesis method is based on intermediate a6 and compound P55, using 4-amino-N-methylpyrazole condensation, with phenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.02(s,1H),10.43(s,1H),8.78(s,1H),8.37(d,J=7.9Hz,1H),8.17(s,1H),7.78(s,1H),7.65(d,J=8.2Hz,1H),7.59(d dd,J=8.2,7.0,1.2Hz,1H),7.44–7.39(m,2H),7.35–7.26(m,3H),7.23–7 .16(m,1H),3.86(s,3H),3.57–3.51(m,2H),3.33–3.27(m,2H).LRMS(ESI + m / z:395.8[M+1] + .

[0352] Example 71: 1-Phenylacetyl-N-(1H-pyrazole-3-substituted)-9H-pyrido[3,4-b]indole-3-amide (P71)

[0353]

[0354] The synthesis method is based on intermediate a6 and compound P55, using 3-aminopyrazole condensation and phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(600MHz,DMSO-d6)δ12.51(s,1H),12.08(s,1H),10.43(s,1H),8.82(s,1 H),8.39(d,J=7.9Hz,1H),7.71(s,1H),7.66(dt,J=8.2,0.9Hz,1H),7.60(dd d,J=8.2,7.0,1.2Hz,1H),7.43–7.39(m,2H),7.35–7.28(m,3H),7.21–7.16( m,1H),6.74(s,1H),3.57–3.51(m,2H),3.28(dd,J=9.5,6.7Hz,2H).LRMS(ESI + m / z:381.9[M+1] + .

[0355] Example 72: N-(1-methyl-4-trifluoromethyl-1H-pyrazole-3-substituted)-1-phenethyl-9H-pyrido[3,4-b]indole-3-amide (P72)

[0356]

[0357] The synthesis method is based on intermediate a6 and compound P55, using 3-amino-4-trifluoromethyl-N-methylpyrazole condensation, with phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(600MHz,DMSO-d6)δ12.12(s,1H),10.38(s,1H),8.80(s,1H),8.50(s,1 H),8.39(d,J=7.9Hz,1H),7.64(dt,J=8.2,1.0Hz,1H),7.59(ddd,J=8.2,6. 9,1.2Hz,1H),7.37–7.33(m,2H),7.32–7.27(m,3H),7.22–7.17(m,1H),3.9 7(s,3H),3.50(dd,J=9.3,6.8Hz,2H),3.29(dd,J=9.3,6.8Hz,2H).LRMS(ESI + m / z: 463.9 [M+1] + .

[0358] Example 73: N-(2,6-dimethylphenyl)-1-(2-methoxyphenethyl)-9H-pyrido[3,4-b]indole-3-amide (P73)

[0359]

[0360] The synthesis method is the same as that of compound P55, using 2-methoxyphenylpropanal as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ11.98(s,1H),9.98(s,1H),8.78(s,1H),8.39(d,J=8.0Hz,1H),7.69–7.55(m,2H),7.36–7.25(m,2H),7.20–7. 10(m,4H),6.94(d,J=8.0Hz,1H),6.84(t,J=7.8Hz,1H),3.75(s,3H),3.51(t,J=7.7Hz,2H),3.41–3.29(m,2H),2.24(s,6H).LRMS(ESI + m / z: 450.2 [M+1] + .

[0361] Example 74: N-(2,6-dimethylphenylethyl)-1-(3-methoxyphenyl)-9H-pyrido[3,4-b]indole-3-amide (P74)

[0362]

[0363] The synthesis method is the same as that of compound P55, using 3-methoxyphenylpropanal as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.00(s,1H),8.76(s,1H),8.38(d,J=7.7Hz,1H),7.70–7.54(m,2H),7.34–7.23( m,3H),7.15(s,3H),6.82(d,J=8.4Hz,2H),3.67(s,3H),3.51(t,J=7.4Hz,2H),3.33–3.22(m,2H),2.24(s,6H).LRMS(ESI + m / z: 450.2 [M+1] + .

[0364] Example 75: N-(2,6-dimethylphenyl)-1-(4-methoxyphenethyl)-9H-pyrido[3,4-b]indole-3-amide (P75)

[0365]

[0366] The synthesis method is based on compound P55, using 4-methoxyphenylpropanal as the substrate aldehyde. 1H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.00(s,1H),8.76(s,1H),8.38(d,J=8.0Hz,1H),7.68–7.55(m,2H),7.33–7.25( m,3H),7.15(s,3H),6.82(d,J=8.6Hz,2H),3.67(s,3H),3.50(t,J=7.7Hz,2H),3.33–3.26(m,2H),2.24(s,6H).LRMS(ESI + m / z: 450.2 [M+1] + .

[0367] Example 76: N-(2,6-dimethylphenyl)-1-(4-hydroxyphenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P76)

[0368]

[0369] The synthesis method is based on compound P55, using 4-hydroxyphenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.08(s,1H),9.13(s,1H),8.77(s,1H),8.38(d,J=8.0Hz,1H),7.69–7.55(m,2H),7. 29(t,J=7.9Hz,1H),7.22–7.11(m,5H),6.66(d,J=8.4Hz,2H),3.53–3.44(m,2H),3.30–3.19(m,2H),2.25(s,6H).LRMS(ESI + m / z: 436.2 [M+1] + .

[0370] Example 77: 1-(4-chlorophenylethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P77)

[0371]

[0372] The synthesis method is the same as that of compound P55, using 4-chlorophenylpropanal as the substrate aldehyde. 1H NMR (400MHz, DMSO-d6) δ12.04(s,1H),10.03(s,1H),8.78(s,1H),8.39(d,J=7.8Hz,1H),7.73–7.54(m,2H),7.41(d ,J=8.4Hz,2H),7.36–7.25(m,3H),7.16(s,3H),3.54(t,J=7.7Hz,2H),3.38(t,J=7.8Hz,2H),2.24(s,6H).LRMS(ESI + m / z:454.2[M+1] + .

[0373] Example 78: N-(2,6-dimethylphenyl)-1-(4-fluorophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P78)

[0374]

[0375] The synthesis method is the same as that of compound P55, using 4-fluorophenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.02(s,1H),10.04(s,1H),8.78(s,1H),8.38(d,J=7.7Hz,1H),7.69–7.56(m,2H),7.41(dd,J=8.2,5.8H z,2H),7.30(t,J=7.4Hz,1H),7.16(s,3H),7.09(t,J=8.8Hz,2H),3.53(t,J=7.7Hz,2H),3.40–3.36(m,2H),2.25(s,6H).LRMS(ESI + m / z: 450.2 [M+1] + .

[0376] Example 79: N-(2,6-dimethylphenyl)-1-(3,5-difluorophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P79)

[0377]

[0378] The synthesis method is the same as that of compound P55, using 3,5-difluorophenylpropanal as the substrate aldehyde. 1H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.06(s,1H),8.78(s,1H),8.38(d,J=7.9Hz,1H),7.70–7.55(m,2H),7.30(t,J=7 .4Hz,1H),7.23–7.10(m,5H),6.99(t,J=9.4Hz,1H),3.55(t,J=7.5Hz,2H),3.44(t,J=7.5Hz,2H),2.24(s,6H).LRMS(ESI + m / z: 456.2 [M+1] + .

[0379] Example 80: N-(2,6-dimethylphenyl)-1-(4-trifluoromethylphenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P80)

[0380]

[0381] The synthesis method is the same as that of compound P55, using 4-trifluoromethylphenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.06(s,1H),10.02(s,1H),8.79(s,1H),8.39(d,J=7.9Hz,1H),7.72–7.54(m ,6H),7.30(t,J=7.5Hz,1H),7.15(s,3H),3.64–3.55(m,2H),3.55–3.44(m,2H),2.23(s,6H).LRMS(ESI + m / z:488.2[M+1] + .

[0382] Example 81: N-(2,6-dimethylphenyl)-1-(4-methylphenethyl)-9H-pyrido[3,4-b]indole-3-amide (P81)

[0383]

[0384] The synthesis method is the same as that of compound P55, using 4-methylphenylpropanal as the substrate aldehyde. 1H NMR(400MHz,Chloroform-d)δ12.06(s,1H),10.06(s,1H),8.83(s,1H),8.66 (d,J=7.6Hz,1H),8.39(d,J=9.6Hz,1H),7.71–7.57(m,1H),7.36–7.20(m,2H ),7.16(s,3H),7.04–6.93(m,1H),6.78(d,J=10.0Hz,1H),6.69(t,J=7.6Hz, 1H),3.74(s,2H),3.29(t,J=7.4Hz,2H),2.25(s,6H),1.99(s,3H).LRMS(ESI + m / z:434.2[M+1] + .

[0385] Example 82: N-(2,6-dimethylphenyl)-1-(4-methylsulfonyloxyphenethyl)-9H-pyrido[3,4-b]indole-3-amide (P82)

[0386]

[0387] The synthesis method is the same as that of compound P55, using 4-methylsulfoxide phenylpropionaldehyde as the substrate aldehyde. 1 H NMR(500MHz,DMSO-d6)δ12.01(s,1H),10.04(s,1H),8.77(s,1H),8.38(d,J=7.9Hz,1H),7.68–7.55(m,6H),7 .30(t,J=7.7Hz,1H),7.15(s,3H),3.64–3.52(m,2H),3.51–3.42(m,2H),2.67(s,3H),2.24(s,6H).LRMS(ESI + m / z: 482.2 [M+1] + .

[0388] Example 83: N-(2,6-dimethylphenyl)-1-(4-cyanophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P83)

[0389]

[0390] The synthesis method is based on compound P55, using 4-cyanophenylpropanal as the substrate aldehyde. 1H NMR (600MHz, DMSO-d6) δ12.03(s,1H),10.00(s,1H),8.78(s,1H),8.38(d,J=7.9Hz,1H),7.76–7.71(m,2H),7.66(d,J=8.2H z,1H),7.62–7.56(m,3H),7.30(t,J=7.4Hz,1H),7.16(s,3H),3.63–3.54(m,2H),3.54–3.44(m,2H),2.23(s,6H).LRMS(ESI + m / z:444.2[M+1] + .

[0391] Synthesis of intermediate I5

[0392]

[0393] The synthesis method is based on compound P55, using 4-bromophenylpropanal as the substrate aldehyde. 1 H NMR (500MHz, DMSO-d6) δ12.11(s,1H),10.02(s,1H),8.77(s,1H),8.38(d,J=7.9Hz,1H),7.74–7.55(m,2H),7. 49–7.41(m,2H),7.39–7.25(m,3H),7.15(s,3H),3.59–3.48(m,2H),3.41–3.31(m,3H),2.24(s,6H).LRMS(ESI + m / z:497.1[M+1] + .

[0394] Example 84: N-(2,6-dimethylphenyl)-1-(4-aminophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P84)

[0395]

[0396] Under argon protection, I5 (124 mg, 0.25 mmol), BocNH (0.5 mmol), Pd2(dba)3 (0.025 mmol), RuPhos (0.025 mmol), and cesium carbonate (244 mg, 0.75 mmol) were added, followed by reflux with toluene overnight (Buchwald-Hartwig reaction). After the reaction was monitored by TLC to confirm its completeness, heating was stopped, and the reaction was allowed to cool to room temperature. The reaction was then quenched with saturated ammonium chloride solution, and the mixture was extracted once with ethyl acetate. The aqueous phase was then extracted four times with dichloromethane until complete extraction. The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated to obtain an oily crude liquid, which was stirred at room temperature for 2 h with 4N hydrochloric acid methanol solution. After evaporation to dryness, the mixture was extracted four times with dichloromethane solution until complete extraction. The organic phases were combined and dried over anhydrous sodium sulfate. The crude liquid was purified by silica gel column chromatography to obtain the target compound as a white solid. 1 H NMR (400MHz, DMSO-d6) δ11.98(s,1H),10.07(s,1H),8.76(s,1H),8.37(d,J=8.7Hz,1H),7.69–7.53(m,2H),7.29(t,J=7.6Hz,1H),7 .15(s,3H),7.04(d,J=8.4Hz,2H),6.48(d,J=8.1Hz,2H),4.81(s,2H),3.50–3.39(m,2H),3.23–3.12(m,2H),2.25(s,6H).LRMS(ESI + m / z:434.2[M+1] + .

[0397] Example 85: N-(2,6-dimethylphenyl)-1-(4-dimethylaminooxyphenethyl)-9H-pyrido[3,4-b]indole-3-amide (P85)

[0398]

[0399] The synthesis method is the same as that of compound P55, using 4-dimethylaminophenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),9.97(s,1H),8.75(s,1H),8.37(d,J=8.0Hz,1H),7.70–7.57(m,2H),7.29(t,J=7.5Hz,1 H),7.23–7.12(m,5H),6.63(d,J=7.6Hz,2H),3.48(t,J=7.6Hz,2H),3.23(t,J=7.8Hz,2H),2.79(s,6H),2.24(s,6H).LRMS(ESI+ m / z:463.2[M+1] + .

[0400] Example 86: N-(2,6-dimethylphenyl)-1-(4-methylsulfonylaminophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P86)

[0401]

[0402] The synthesis method is the same as that of compound P55, using 4-methylsulfonylaminophenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.06(s,1H),10.14(s,1H),8.81(s,1H),8.39(d,J=7.9Hz,1H),7.67–7.58(m,2H),7.55(d,J=8.4Hz,2H),7 .45(d,J=8.3Hz,2H),7.31(t,J=7.4Hz,1H),7.16(s,3H),3.59–3.54(m,2H),3.50(s,3H),3.47–3.43(m,2H),2.26(s,6H).LRMS(ESI + m / z:513.2[M+1] + .

[0403] Example 87: N-(2,6-dimethylphenyl)-1-(4-acetamidophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P87)

[0404]

[0405] The synthesis method is the same as that of compound P55, using 4-acetamidophenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),10.04(s,1H),9.81(s,1H),8.76(s,1H),8.38(d,J=8.1Hz,1H),7.69–7.54(m,2H),7.46( d,J=7.8Hz,2H),7.29(d,J=8.4Hz,3H),7.15(s,3H),3.57–3.47(m,2H),3.34–3.26(m,2H),2.24(s,6H),2.00(s,3H).LRMS(ESI + m / z: 477.2 [M+1] + .

[0406] Example 88: N-(2,6-dimethylphenyl)-1-(4-dimethylphosphorylphenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P88)

[0407]

[0408] The synthesis method is based on compound P55, using 4-dimethylphosphorylphenylpropionaldehyde as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.05(s,1H),10.06(s,1H),8.78(s,1H),8.38(d,J=8.5Hz,1H),8.03–7.83(m,1H) ,7.79–7.36(m,9H),3.66–3.52(m,2H),3.51–3.42(m,2H),2.89(s,3H),2.73(s,3H),2.25(s,6H).LRMS(ESI + m / z: 496.2 [M+1] + .

[0409] Example 89: N-(2,6-dimethylphenyl)-1-(2-(pyridin-4-substituted)ethyl)-9H-pyrido[3,4-b]indole-3-amide (P89)

[0410]

[0411] The synthesis method is the same as that of compound P55, using 4-propanoylpyridine as the substrate aldehyde. 1 H NMR(400MHz,DMSO-d6)δ12.08(s,1H),10.06(s,1H),8.78(s,1H),8.51–8.34(m,3H),7.71–7.56(m,2H),7.41(d,J =4.3Hz,2H),7.30(t,J=7.8Hz,1H),7.16(s,3H),3.64–3.54(m,2H),3.44(t,J=7.7Hz,2H),2.23(s,6H).LRMS(ESI + m / z:421.2[M+1] + .

[0412] Example 90: N-(2,6-dimethylphenyl)-1-(2-(pyridin-3-substituted)ethyl)-9H-pyrido[3,4-b]indole-3-amide (P90)

[0413]

[0414] The synthesis method is the same as that of compound P55, using 3-propanoylpyridine as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.03(s,1H),10.08(s,1H),8.78(s,1H),8.59(s,1H),8.42–8.33(m,2H),7.78(d,J=7.9Hz,1H),7 .69–7.56(m,2H),7.29(t,J=8.7Hz,2H),7.16(s,3H),3.56(t,J=7.5Hz,2H),3.42(t,J=7.7Hz,2H),2.24(s,6H).LRMS(ESI + m / z:421.2[M+1] + .

[0415] Example 91: 4-(2-(3-((2,6-dimethylphenyl)amide)-9H-pyrido[3,4-b]indole-1-substituted)ethyl)benzoic acid (P91)

[0416]

[0417] The synthesis method is the same as that of compound P55, using 4-propanoylbenzoic acid as the substrate aldehyde. 1 H NMR (400MHz, DMSO-d6) δ12.05(s,1H),10.03(s,1H),8.77(s,1H),8.38(d,J=7.7Hz,1H),7.85(d,J=7.8Hz,2H),7. 72–7.45(m,4H),7.30(t,J=7.6Hz,1H),7.15(s,3H),3.67–3.53(m,2H),3.53–3.41(m,2H),2.23(s,6H).LRMS(ESI + m / z: 464.2 [M+1] + .

[0418] Example 92: N-(2,6-dimethylphenyl-1-(4-(4-methylpiperazin-1-carbonyl)phenethyl)-9H-pyrido[3,4-b]indole-3-amide (P92)

[0419]

[0420] The synthesis method is based on compound P55, using 4-propanoyl-N-methylbenzoylpiperazine as the substrate aldehyde. 1H NMR (600MHz, DMSO-d6) δ11.99(s,1H),10.09(s,1H),8.77(s,1H),8.38(d,J=7.9Hz,1H),7.66–7.57(m,2H),7.45(d ,J=8.1Hz,2H),7.31–7.27(m,3H),7.16(s,3H),3.61–3.50(m,4H),3.48–3.39(m,4H),2.30–2.19(m,13H).LRMS(ESI + m / z: 546.3 [M+1] + .

[0421] Example 93: N-(2,6-dimethylphenyl)-1-(4-((4-methylpiperazine-1-substituted)sulfonyl)phenethyl)-9H-pyrido[3,4-b]indole-3-amide (P93)

[0422]

[0423] The synthesis method is based on compound P55, using 4-propanal-N-methylbenzylpiperazine as the substrate aldehyde. 1 HNMR (400MHz, DMSO-d6) δ12.04(s,1H),10.10(s,1H),8.78(s,1H),8.38(d,J=8.0Hz,1H),7.79–7.52(m,6H),7.35–7. 25(m,1H),7.16(s,3H),3.66–3.46(m,4H),2.97–2.69(m,4H),2.47–2.30(m,4H),2.24(s,6H),2.16(s,3H).LRMS(ESI + m / z:582.2[M+1] + .

[0424] Example 94: N-(2,6-dimethylphenyl)-1-(4-((4-methylpiperazine-1-substituted)methyl)phenethyl)-9H-pyrido[3,4-b]indole-3-amide (P94)

[0425]

[0426] The synthesis method is based on compound P55, using 1-methyl-4-(4-propanal-benzyl)piperazine as the substrate aldehyde. 1HNMR (600MHz, DMSO-d6) δ12.00(s,1H),9.99(s,1H),8.76(s,1H),8.37(d,J=7.9Hz,1H),7.66–7.62(m,1H),7.59(t,J= 7.7Hz,1H),7.36–7.26(m,3H),7.19–7.12(m,5H),3.55–3.49(m,2H),3.44–3.37(m,2H),2.33–2.13(m,19H).LRMS(ESI + m / z:532.3[M+1] + .

[0427] Example 95: N-(2,6-dimethylphenyl)-1-(4-morpholinophenylethyl)-9H-pyrido[3,4-b]indole-3-amide (P95)

[0428]

[0429] The synthesis method is the same as that of compound P55, using p-methylsulfonylphenylpropionaldehyde as the substrate aldehyde. 1 H NMR (600MHz, DMSO-d6) δ12.04(s,1H),10.08(s,1H),8.78(s,1H),8.38(d,J=7.9Hz,1H),7.88–7.80(m,2H),7.71–7.56 (m,4H),7.30(t,J=7.4Hz,1H),7.16(s,3H),3.61–3.55(m,2H),3.55–3.49(m,2H),3.15(s,3H),2.24(s,6H).LRMS(ESI + m / z:498.2[M+1] + .

[0430] Example 96: 1-(4-(4-acetylpiperazine-1-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P96)

[0431]

[0432] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using N-Boc piperazine as the substrate amine. The Boc protecting group was then removed by stirring at room temperature for 2 hours with a 5:1 volume ratio of dichloromethane / trifluoroacetic acid. After evaporation to dryness, an equivalent amount of acryloyl chloride and excess triethylamine were added to dichloromethane as solvent, and the mixture was reacted at room temperature for 1 hour before evaporation to dryness. Silica gel column chromatography yielded an off-white solid (overall yield 42%). 1H NMR(600MHz,DMSO-d6)δ12.00(s,1H),9.94(s,1H),8.75(s,1H),8.37(d,J=7.9Hz,1H), 7.68–7.63(m,1H),7.59(t,J=7.7Hz,1H),7.29(t,J=7.7Hz,1H),7.27–7.21(m,2H),7.1 5(s,3H),6.89–6.79(m,3H),6.14(d,J=19.0Hz,1H),5.71(d,J=12.8Hz,1H),3.70–3.59 (m,4H),3.54–3.45(m,2H),3.30–3.22(m,2H),3.06–2.96(m,4H),2.23(s,6H).LRMS(ESI + m / z:558.3[M+1] + .

[0433] Example 97: N-(2,6-dimethylphenyl)-1-(4-(4-(4-(methylsulfonyl)piperazine-1-substituted)phenethyl)-9H-pyrido[3,4-b]indole-3-amide (P97)

[0434]

[0435] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and N-methylsulfonylpiperazine as the substrate amine. The product was an off-white solid (yield 56%). 1 H NMR (600MHz, DMSO-d6) δ12.00(s,1H),9.98(s,1H),8.76(s,1H),8.37(d,J=7.9Hz,1H),7.68–7.56(m,2H),7.32–7.22(m,3H),7.15(s,3H) ),6.88(d,J=8.4Hz,2H),3.54–3.45(m,2H),3.31–3.24(m,2H),3.24–3.18(m,4H),3.16–3.08(m,4H),2.91(s,3H),2.24(s,6H).LRMS(ESI + m / z:582.2[M+1] + .

[0436] Example 98: 1-(4-(4-(cyclopropylacyl)piperazine-1-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P98)

[0437]

[0438] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and cyclopropionylpiperazine as the substrate amine. The product was an off-white solid (yield 45%). 1 H NMR (600MHz, DMSO-d6) δ12.01(s,1H),9.95(s,1H),8.76(s,1H),8.37(d,J=7.9Hz,1H),7. 68–7.62(m,1H),7.59(t,J=7.6Hz,1H),7.33–7.21(m,3H),7.15(s,3H),6.87(d,J=8.5Hz,2 H),3.82–3.73(m,2H),3.61–3.54(m,2H),3.50(t,J=7.8Hz,2H),3.26(t,J=7.8Hz,2H),3. 09–3.03(m,2H),3.00–2.96(m,2H),2.24(s,6H),2.02–1.97(m,1H),1.96(s,4H).LRMS(ESI + m / z: 572.3 [M+1] + .

[0439] Example 99: 1-(4-(4-cyclopropylpiperazine-1-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P99)

[0440]

[0441] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and N-methylpiperazine as the substrate amine, resulting in an off-white solid (yield 26%). 1 H NMR(600MHz,DMSO-d6)δ12.00(s,1H),9.99(s,1H),8.75(s,1H),8.37(d,J =7.9Hz,1H),7.67–7.57(m,2H),7.29(t,J=7.4Hz,1H),7.22(d,J=8.5Hz,2H ),7.15(s,3H),6.83(d,J=8.6Hz,2H),3.52–3.46(m,2H),3.28–3.22(m,2H) ,3.07–2.98(m,4H),2.48–2.40(m,4H),2.24(s,6H),2.22(s,3H).LRMS(ESI + )m / z:518.3[M+1] + .

[0442] Example 100: 1-(4-(4-(dimethylaminoacetyl)piperazine-1-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P100)

[0443]

[0444] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and 2-dimethylamino-1-(piperazin-1-substituted)ethane-1-one as the substrate amine. The product was a white solid (yield 44%). 1 H NMR (600MHz, DMSO-d6) δ12.00(s,1H),9.95(s,1H),8.75(s,1H),8.37(d,J=7.9Hz, 1H),7.65(d,J=8.2Hz,1H),7.59(t,J=7.7Hz,1H),7.29(t,J=7.4Hz,1H),7.24(d,J =8.5Hz,2H),7.15(s,3H),6.86(d,J=8.5Hz,2H),3.62–3.53(m,4H),3.50(t,J=7.7 Hz,2H),3.28–3.24(m,4H),3.05–2.96(m,4H),2.27(s,6H),2.23(s,6H).LRMS(ESI + m / z: 589.3 [M+1] + .

[0445] Example 101: 1-(4-(4-acetyl-3-methylpiperazine-1-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P101)

[0446]

[0447] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and 1-(2-methylpiperazine-1-substituted)ethane-1-one as the substrate amine. The product was a white solid (yield 51%). 1H NMR (600MHz, DMSO-d6) δ12.00(s,1H),9.96(s,1H),8.75(s,1H),8.37(d,J=7.9Hz,1H),7.68–7.56(m,2H),7.29(t,J=7.4Hz,1H),7.24(d,J =8.6Hz,2H),7.18–7.14(m,3H),6.83(d,J=8.0Hz,2H),3.53–3.46(m,2H),3.29–3.23(m,2H),2.23(s,6H),2.02(d,J=19.3Hz,3H).LRMS(ESI + m / z: 560.3 [M+1] + .

[0448] Example 102: 1-(4-(5-acetyl-2,5-bicyclo[2.2.2]octane-2-substituted)phenethyl)-N-(2,6-dimethylphenyl)-9H-pyrido[3,4-b]indole-3-amide (P102)

[0449]

[0450] The synthesis method was based on the Buchwald-Hartwig reaction of compound P84, using intermediate I5 as the starting material and 1-(2,5-diazabibridged[2.2.2]octane-2-substituted)ethane-1-one as the substrate amine, resulting in a white solid (yield 48%). 1 HNMR(600MHz,DMSO-d6)δ11.99(s,1H),10.00(s,1H),8.76(s,1H),8.37(d,J=7.9Hz,1H),7.69–7.63(m,1H),7.59(t,J=7.6Hz,1H),7.29(t,J=7.4H z,1H),7.22–7.11(m,5H),6.57(dd,J=8.5,6.2Hz,2H),3.52–3.41(m,5H), 3.29–3.19(m,5H),2.24(s,6H),2.01(s,3H),1.88–1.68(m,4H).LRMS(ESI + m / z: 572.3 [M+1] + .

[0451] Test Implementation Examples

[0452] 1. Main experimental materials and instruments

[0453] Materials: Fetal bovine serum, α-MEM medium, and penicillin / streptomycin were purchased from Gibco; DMSO, MTT, and TRAP staining kits were purchased from Sigma; cytokines such as mM-CSF, mRANKL, hRNAK, and hRNAKL were purchased from Peprotech; cell lysis buffer was purchased from Promega; PBS was purchased from Wisent; and 4-week-old C57BL / 6 mice were purchased from Slack.

[0454] Cells: Bone marrow osteoclast precursor cells, obtained from femoral and tibial medullary cavity cells of C57BL / 6 mice under normal culture conditions, cultured in complete α-MEM medium, i.e., α-MEM + 10% fetal bovine serum + 1% penicillin / streptomycin.

[0455] Instruments: Thermo Scientific CO2 incubator, Olympus inverted microscope, Tecan microplate reader, GE Biacore T200.

[0456] 2. Experimental methods:

[0457] 1) Preparation of test cells:

[0458] C57BL / 6 mice were euthanized by cervical dislocation, disinfected by immersion in 75% alcohol, and under aseptic conditions, the long bones of the hind limbs (femur and tibia) were dissected, and attached soft tissue was removed. The surface of the bone marrow cavity was repeatedly flushed with complete culture medium to completely remove the cells. The cell suspension was filtered through a cell strainer, and after quantitative analysis, the cells were seeded into 10cm cell culture dishes and cultured overnight under 5% CO2 and saturated humidity. The next day, the supernatant of non-adherent cells was collected by centrifugation and replaced with fresh complete proliferation medium (complete medium with 30 ng / mL M-CSF). Culture for another two days to obtain bone marrow osteoclast precursor cells. Two days later, the bone marrow osteoclast precursor cells were seeded into well plates at a specific concentration and induction medium (complete medium with 30 ng / mL M-CSF and 50 ng / mL RANKL) was added. Culture for 5-6 days to obtain mature osteoclasts.

[0459] 2) CCK-8 (Cell Counting Kit-8) assay to detect the effect of compounds on bone marrow osteoclast progenitor cell survival:

[0460] Bone marrow osteoclast precursor cells were seeded at a concentration of 5000 cells per well (100 μL per well) into 96-well plates and cultured overnight. An experimental group and a negative control group were included. Experimental group: A carboline compound solution was prepared to a final concentration of 10 μM using proliferation medium and added to the wells of the experimental group. Negative control group: Proliferation medium containing the same concentration of DMSO was added to the experimental group. After 48 h of culture, 100 μL of CCK-8 was added to each well, and the mixture was incubated at 37°C for 1 h. The absorbance at 450 nm was then measured using a microplate reader to calculate cell viability. The inhibition rate was calculated as follows:

[0461]

[0462] 3) TRAP staining assay to detect the compound's inhibition of osteoclast formation:

[0463] Based on the results of the CCK-8 toxicity assay, compounds that were non-toxic to bone marrow osteoclast precursor cells were selected for TRAP staining experiments.

[0464] Bone marrow osteoclast precursor cells were seeded at a concentration of 5000 cells per well in 96-well plates with 100 μL of culture medium per well and cultured overnight. The next day, the cells were treated differently. The experimental group cells were treated as follows: carboline compounds (compounds I-1 to I-41) were prepared into 5 μM solutions using induction medium and added to the experimental group wells. The negative control group was treated with the same concentration of DMSO induction medium as the experimental group, and the blank control group was treated with the same concentration of DMSO proliferation medium. The stimulated cells were pre-incubated for 4 days in a 5% CO2, 37°C incubator, with the medium changed every 2 days (old medium removed, 100 μL of fresh medium containing the same concentration of the drug added). After 4 days, the osteoclasts bleed out, were washed three times with PBS, fixed with 4% paraformaldehyde for 10 minutes, washed once with PBS, and air-dried. The cells were then treated with 0.8 mol·L⁻¹ of DMSO. -1 pH 5.0 sodium acetate buffer, 0.1 g / L -1 Naphthol AS-BI phosphate, 0.6 g / L -1 Grenache red GBC salt, 0.2 mol·L⁻¹ -1 The cells were reacted in tartaric acid solution for 60 minutes, followed by gradient dehydration with ethanol. Osteoclast-like cells with ≥3 nuclei were counted. The inhibition rate was calculated as follows:

[0465]

[0466] 3. Results:

[0467] (1) Study on the survival effect of carboline compounds of the present invention on bone marrow osteoclast precursor cells

[0468] 10 μM of the carboline compound of this invention was co-incubated with bone marrow osteoclast precursor cells, and cell viability was detected by CCK-8 assay. Some results are shown in Table 1.

[0469] Table 1. Effects of carboline compounds on bone marrow osteoclast progenitor cell survival

[0470]

[0471]

[0472] (2) Study on the inhibitory effect of the carboline compounds of the present invention on osteoclasts

[0473] Based on the results of the CCK-8 experiment, 5 μM of the non-toxic carboline compound of the present invention was selected for the experiment. Osteoclast differentiation of bone marrow osteoclast precursor cells was induced by M-CSF and RANKL. The effect of carboline compound on osteoclast differentiation process was detected by TRAP staining experiment. Some results are shown in Table 2.

[0474] The results show that some of the carboline compounds of the present invention have a very good inhibitory effect on the differentiation process of bone marrow osteoclasts.

[0475] Table 2. Inhibitory effects of carboline compounds on bone marrow osteoclast differentiation

[0476]

Claims

1.A 3-amido-β-carboline compound represented by the general formula P-2, or a pharmaceutically acceptable salt thereof, wherein R1 represents hydrogen; R2and R3are each independently hydrogen, substituted or non-substituted C6-C 10 aryl, substituted or non-substituted 5-8 membered heteroaryl, substituted or non-substituted 3-8 membered heterocyclyl, substituted or non-substituted C3-C6cycloalkyl; R5is substituted or unsubstituted C6-Ci8aryl, substituted or unsubstituted 5-8 membered heteroaryl; 10 R5is substituted or unsubstituted C6-Ci8aryl, substituted or unsubstituted 5-8 membered heteroaryl; the substituent in the above "substituted or unsubstituted" means a substituent selected from the group consisting of halogen, hydroxyl, amino, C1-C6 alkyl. 2.A 3-amido-β-carboline compound, or a pharmaceutically acceptable salt thereof, wherein, the 3-amido-β-carboline compound is selected from the group consisting of the following compounds: 。 3.A method for preparing a 3-amido-β-carboline compound represented by the general formula P-c, which is a method comprising the following steps: Step i: condensation of the starting material with an organic amine under the action of a condensing agent to obtain an amide compound a9; Step ii: deprotection of the compound a9 under the action of hydrogen chloride or trifluoroacetic acid to obtain a corresponding salt, which is directly used in the next step; Step iii: condensation and ring closure of the corresponding salt with substituted or unsubstituted arylpropanal to obtain an intermediate a10; Step iv: oxidative aromatization of the compound a10 with N-chlorosuccinimide or 2,3-dichloro-5,6-dicyano-1,4-benzoquinone to obtain a 3-amido-β-carboline compound represented by the general formula P-c; wherein R1 to R3, R5 have the same meanings as R1 to R3, R5 in claim 1. 4.A pharmaceutical composition comprising a therapeutically effective amount of one or more selected from the group consisting of a 3-amido-β-carboline compound according to any one of claims 1-2, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 5.Use of a 3-amido-β-carboline compound according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 4 in the preparation of a medicament for treating osteoporosis.

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