PRMT5 inhibitor, and composition and use thereof

Abstract

Provided are a PRMT5 inhibitor, and a composition and the use thereof. Specifically, the present invention relates to a PRMT5 inhibitor, and a stereoisomer thereof or a pharmaceutically acceptable salt thereof. The heterocyclic compound that inhibits PRMT5 activity has a structure as represented by formula (I). Further provided are a composition containing the compound and the use thereof as a PRMT5 inhibitor.

Description

A class of PRMT5 inhibitors, their compositions and uses Technical Field

[0001] This invention belongs to the field of medicinal chemistry, specifically relating to a class of PRMT5 inhibitor compounds and their stereoisomers, or pharmaceutically acceptable salts thereof, and to pharmaceutical compositions comprising such compounds as active ingredients and their pharmaceutical use in the treatment of diseases, symptoms, syndromes or disorders related to the PRMT5 enzyme. Background Technology

[0002] Arginine methylation, a type of histone methylation, is one of the most common post-translational modifications in mammals and is primarily regulated by the PRMT gene family. Arginine methyltransferases (PRMTs) regulate arginine methylation in three different ways: monomethylarginine (MMA), asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA) methylation. There are nine PRMTs in mammals, classified into three types: type I (PRMT1, 2, 3, 4, 6, 8; primarily catalyzing the production of MMA and ADMA), type II (PRMT5, 9; primarily catalyzing the production of MMA and SDMA), and type III (PRMT7; primarily catalyzing the production of MMA). PRMTs can methylate histones and various non-histone proteins. Studies have shown that alterations in PRMT enzyme activity, gene mutations, or deletions often lead to developmental abnormalities in animals and also induce the occurrence, development, or metastasis of cancer. Among the many PRMTs, PRMT5, in particular, has received increasing attention due to its increasingly confirmed role in cancer.

[0003] Protein arginine methyltransferase 5 (PRMT5) is a typical type II methyltransferase that transfers a methyl group from SAM to the two ω-guanidino nitrogen atoms of arginine, resulting in ω-NG, NG-disymmetric methylation of the protein substrate. It is located in the nucleus and cytoplasm and performs different functions by modifying histones or non-histone proteins. As an epigenetic enzyme, PRMT5's most intriguing feature is its "synthetic lethality" mechanism.

[0004] Synthetic lethality means that if either of two genes in a cell mutates alone or does not function, the cell will not die; however, if both genes mutate simultaneously or are not expressed, the cell will die. This principle can be used to selectively kill tumor cells without affecting normal somatic cells.

[0005] In 2016, two articles published in *Science* reported for the first time the "synthetic lethal" effect of inhibiting PRMT5 in MTAP-deficient tumors. The gene responsible for synthetic lethality with PRMT5 is methionine phosphorylase (MTAP), a tumor suppressor gene that is commonly deleted in tumors. MTAP-deficient tumors account for approximately 15% of all solid tumors, including about 15% of non-small cell lung cancer (NSCLC), 28% of esophageal cancer, 26% of bladder cancer, and 10% of esophageal and gastric cancer.

[0006] Besides its synthetic lethal effects, PRMT5 also participates in DNA repair, cell cycle, and transcriptional regulation through methylation modification of its substrate arginine, playing a crucial role in various cellular functions. Currently, research on small molecule inhibitors of PRMT5 has become a hot topic in anti-tumor drug development.

[0007] The results of most first-generation PRMT5 inhibitor clinical trials were generally unsatisfactory: Clinical studies showed that GSK-3326595 responded to multiple tumor types. However, 89% of participants reported adverse events, including anemia, thrombocytopenia, neutropenia, and fatigue. A Phase I clinical trial of JNJ-64619178 showed that intermittent dosing maintained target inhibition in cancer patients. Conversely, JNJ-64619178 showed limited / no efficacy in patients with myelodysplastic syndromes. Most patients treated with JNJ-64619178 experienced adverse events, including thrombocytopenia, anemia, and nausea. PF-06939999, as a PRMT5 inhibitor, showed an objective tumor response in patients with squamous cell carcinoma of the neck and non-small cell lung cancer. Cytopenia was observed in clinical trials; these were dose-dependent and reversible with dose correction. Clinical trials of the aforementioned PRMT5 inhibitors appear to have been halted.

[0008] In fact, aside from Prelude Therapeutics' projects, most active programs focus on second-generation PRMT5 inhibitors, specifically PRMT5-MTA inhibitors. In cancer cells lacking MTAP, MTA replaces SAM in binding to PRMT5, forming an inactive PRMT5-MTA complex. Second-generation PRMT5 inhibitors bind to the PRMT5-MTA complex, killing MTAP-deficient cancer cells while preserving normal cells. This property allows second-generation PRMT5 inhibitors to target MTAP-deficient cancer cells more specifically and may reduce the incidence of adverse events.

[0009] Currently, there are second-generation PRMT5 inhibitors in Phase I / II clinical trials, including MRTX-1719, SKL-27969, TNG-908, TNG-462, and AMG-193, but none of them have been officially launched. There is an urgent need to develop PRMT5 inhibitors with better efficacy and better selectivity for MTAP-deficient cancer cell lines. Summary of the Invention

[0010] Invention Overview

[0011] The purpose of this invention is to provide a class of novel PRMT5 inhibitor compounds with novel structure, good pharmacodynamic activity, superior selectivity (especially HCT116 WT), and better pharmacokinetic properties, as well as their stereoisomers and their use in the treatment of cancer.

[0012] The present invention solves the above-mentioned technical problems through the following technical solutions.

[0013] On the one hand, the present invention provides compounds as shown in Formula I and their stereoisomers, or pharmaceutically acceptable salts thereof:

[0014] R1, R2, and R3 are each independently selected from H, halogen, -C1-C6 alkyl, -C3-C6 cycloalkyl, -C1-C6 haloalkyl, and -OR. a1 -NR a2 R a3 ;

[0015] Alternatively, a 5-10 membered heterocyclic group or a 5-10 membered heteroaryl group may be formed between R2, R3 and the connected C, wherein the 5-10 membered heterocyclic group or the 5-10 membered heteroaryl group may optionally be surrounded by one or more halogen groups, -C1-C6 alkyl groups, -C3-C6 cycloalkyl groups, -C1-C6 haloalkyl groups, or -OR groups. a1 -NR a2 R a3 replace.

[0016] R a1 R a2 R a3 Each is independently selected from H, -C1-C6 alkyl, -C1-C6 haloalkyl, and -C3-C6 cycloalkyl;

[0017] R4 is selected from H, -C1-C6 alkyl;

[0018] R5, R6, and R7 are each independently selected from H, halogen, -C1-C6 alkyl, -C1-C6 alkoxy, and -C1-C6 haloalkyl;

[0019] A1 and A2 are each independently selected from -NR8 and -CR9R. 10 ;

[0020] R8 is selected from H, -C1-C6 alkyl, -C1-C6 alkoxy, -C1-C6 haloalkyl, -C3-C6 cycloalkyl, and -(CH2). r OC1-C6 alkyl, -C1-C6 hydroxyalkyl, -(CH2) r -NR a2 R a3 -C(O)C1-C6 alkyl;

[0021] R9, R 10 Each is independently selected from H, -C1-C6 alkyl groups;

[0022] m is 0, 1, 2, 3; n is 0, 1, 2, 3; r is 0, 1, 2, 3, 4.

[0023] As a preferred technical solution, the 5-10 membered heterocyclic group and the 5-10 membered heteroaryl group are selected from 5-6 membered heterocyclic groups and 5-6 membered heteroaryl groups.

[0024] As a preferred technical solution, the compound is selected from:

[0025] As a preferred technical solution, the compound is selected from:

[0026] As a preferred technical solution, R1 is selected from H, halogen, C1-C6 alkyl, and C3-C6 cycloalkyl.

[0027] As a more preferred technical solution, R1 is selected from H, methyl, chlorine, and cyclopropyl.

[0028] As a preferred technical solution, R2 and R3 are each independently selected from H, amino, ethyl, methyl, trifluoromethyl, cyclopropyl, methoxy, trifluoromethoxy, chlorine, and trifluoromethoxy.

[0029] As a preferred technical solution, R4 is selected from -C1-C6 alkyl groups.

[0030] As a more preferred technical solution, R4 is selected from methyl.

[0031] As a preferred technical solution, m is 1.

[0032] As a preferred technical solution, R5 and R6 are each independently selected from H and -C1-C6 alkyl groups.

[0033] As a more preferred technical solution, R5 and R6 are selected from H.

[0034] As a preferred technical solution, R7 is selected from H, -C1-C6 alkyl groups.

[0035] As a more preferred technical solution, R7 is selected from H and methyl.

[0036] As a preferred technical solution, R8 is selected from methyl, ethyl, cyclopropyl, isopropyl, -CH2CH2F, -CH2CHF2, -(CH2)2OCH3, -(CH2)2OH, -(CH2)2N(CH3)2, -C(O)CH3, -CH2CH2OCH3.

[0037] The present invention also provides compounds selected from the following formulas and their stereoisomers, or pharmaceutically acceptable salts thereof:

[0038] The present invention also provides the compounds described in any of the above claims, their stereoisomers or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers.

[0039] The present invention also provides the use of any of the compounds described above, their stereoisomers, or pharmaceutically acceptable salts thereof, in the preparation of medicaments for treating diseases, symptoms, syndromes, or disorders associated with PRMT5.

[0040] As a preferred technical solution, the diseases, symptoms, syndromes, or disorders related to the PRMT5 enzyme include cancer, blood diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, genetic diseases, hormone-related diseases, immunodeficiency diseases, cell death-related diseases, destructive bone diseases, thrombin-induced platelet aggregation, liver diseases, and cardiovascular diseases. Preferably, the PRMT5 enzyme-related diseases, symptoms, syndromes, or disorders are cancer; more preferably, the cancer is advanced solid tumors, metastatic pancreatic cancer, or metastatic non-small cell lung cancer. Lung cancer, neuro-oncology, adenocarcinoma, endometrioid carcinoma, metastatic esophageal cancer, metastatic head and neck cancer, squamous cell carcinoma, cervical cancer, myelodysplastic syndrome, non-Hodgkin's lymphoma, acute myeloid leukemia, adenoid tumors, hematologic malignancies, melanoma, pancreatic tumors, brain tumors, glioblastoma, glioma, myelofibrosis, breast tumors, chronic myelomonocytic leukemia, diffuse large B-cell lymphoma, bladder cancer, bile duct cancer, mesothelioma, ovarian cancer, lung cancer, prostate cancer, colon cancer, stomach cancer, esophageal cancer, and hepatocellular carcinoma.

[0041] The present invention also provides a method for treating MTAP deficiency and / or MTA accumulation disease in subjects in need with any of the compounds or pharmaceutical compositions described above.

[0042] As a preferred technical solution, the disease is a proliferative disease.

[0043] As a more preferred technical solution, the proliferative disease is MTAP deficiency and / or MTA-accumulated cancer.

[0044] As a further preferred technical solution, the cancer is glioblastoma or malignant peripheral nerve sheath tumor.

[0045] (MPNST), esophageal cancer (e.g., esophageal squamous cell carcinoma or esophageal adenocarcinoma), bladder cancer (e.g., bladder urothelial carcinoma), pancreatic cancer (e.g., pancreatic adenocarcinoma), mesothelioma, melanoma, non-small cell lung cancer (NSCLC; e.g., lung squamous or lung adenocarcinoma), astrocytoma, undifferentiated pleomorphic sarcoma, diffuse large B-cell lymphoma (DLBCL), leukemia, head and neck cancer, gastric adenocarcinoma, myxoid fibrosarcoma, cholangiosarcoma, brain cancer, gastric cancer, kidney cancer, breast cancer, endometrial cancer, urinary tract cancer, liver cancer, soft tissue cancer, pleural cancer, and colorectal cancer or sarcoma.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] This invention utilizes computer-aided design technology to modify analogs such as TNG908 and TNG462. Through computer simulation, molecular docking analysis, and structure-activity relationship studies, a variety of novel compounds were screened. This series of compounds can bind to the PRMT5-MTA complex and inhibit PRMT5 function, exhibiting extremely strong PRMT5 inhibitory activity. In vitro and in vivo experiments further demonstrate that this series of compounds can prevent and / or treat diseases such as cancer, hematological diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, genetic diseases, hormone-related diseases, immunodeficiency diseases, cell death-related diseases, destructive bone diseases, thrombin-induced platelet aggregation, liver diseases, and cardiovascular diseases. They exhibit good activity, strong inhibitory activity against MATP-deficient HCT116 tumor cells, weak inhibitory effect on HCT116 WT cells, and remarkable selectivity, with even better pharmacokinetic properties in rats.

[0048] Invention Details

[0049] The various aspects and features of the present invention will be further described below.

[0050] The compounds of this invention have an asymmetric center. Compounds containing asymmetric substitution atoms in this invention can be isolated into optically active or racemic forms. Those skilled in the art know how to prepare the optically active forms, for example, through racemic resolution or synthesis from optically active starting materials. Unless otherwise specified, this invention includes all chiral, diastereomers, and racemates. Methods for preparing the compounds of this invention and their intermediates are part of this invention. All tautomers of the compounds of this invention are also part of this invention.

[0051] "alkyl" refers to a group having 1 to 10 carbon atoms, either straight-chain or branched, saturated hydrocarbon groups ("C1-C10 alkyl"). In some embodiments, the alkyl group has 1 to 8 carbon atoms ("C1-C8 alkyl"). In some embodiments, the alkyl group has 1 to 6 carbon atoms ("C1-C6 alkyl"), 1 to 5 carbon atoms ("C1-C5 alkyl"), 1 to 4 carbon atoms ("C1-C4 alkyl"), 1 to 3 carbon atoms ("C1-C3 alkyl"), 1 to 2 carbon atoms ("C1-C2 alkyl"), or 1 carbon atom ("C1 alkyl"). In some embodiments, the alkyl group has 2 to 6 carbon atoms ("C2-C6 alkyl"). Examples of C1-C6 alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8), etc. Each alkyl group may be optionally substituted independently, i.e., unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents; for example, substituted with 1 to 5 substituents, 1 to 3 substituents, or substituted with 1 substituent (“substituted alkyl”). In some embodiments, the alkyl group is an unsubstituted C1–C10 alkyl group (e.g., –CH3). In some embodiments, the alkyl group is a substituted C1–C6 alkyl group. Commonly used alkyl abbreviations include Me (–CH3), Et (–CH2CH3), iPr (–CH(CH3)2), etc.

[0052] As described herein, the terms “halogen,” “halogenated,” “halogenated,” etc., refer to fluorine, chlorine, bromine, or iodine, particularly fluorine, chlorine, and bromine, with fluorine and chlorine being especially preferred.

[0053] "Haloalkyl" refers to an alkyl group as described herein (e.g., C1-C6 alkyl) in which one or more hydrogen atoms are replaced by a halogen (e.g., monohaloalkyl, dihaloalkyl, and trihaloalkyl). Such groups include, but are not limited to, chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chloro-fluoroalkyl, chloro-difluoroalkyl, and 2-fluoroisobutyl.

[0054] "Alkoxy" refers to an alkyl group (e.g., C1-C6 alkyl) as described herein, which is attached to the molecule by an oxygen atom. This includes portions in which the alkyl moiety can be straight-chain or branched, such as methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

[0055] In some embodiments, "cycloalkyl" is a monocyclic saturated cycloalkyl group having 3 to 10 ring carbon atoms ("C3-10 cycloalkyl" or "C3-C10 cycloalkyl"). In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms ("C3-8 cycloalkyl" or "C3-C8 cycloalkyl"). In some embodiments, the cycloalkyl group has 3 to 6 ring carbon atoms ("C3-6 cycloalkyl" or "C3-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms ("C5-6 cycloalkyl" or "C5-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 10 ring carbon atoms ("C5-10 cycloalkyl" or "C5-C10 cycloalkyl"). Examples of C5-C6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-C6 cycloalkyl groups include the aforementioned C5-C6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-C8 cycloalkyl groups include the aforementioned C3-C6 cycloalkyl groups, as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each example of a cycloalkyl group is independently unsubstituted (“unsubstituted cycloalkyl”) or substituted with one or more substituents (“substituted cycloalkyl”). In some embodiments, the cycloalkyl group is an unsubstituted C3-C10 cycloalkyl group. In some embodiments, the cycloalkyl group is a substituted C3-C10 cycloalkyl group.

[0056] The terms "heterocyclic" and "heterocyclic group" are used interchangeably and refer to substituted and unsubstituted 3- to 7-membered monocyclic groups, 7- to 11-membered bicyclic groups, and 10- to 15-membered tricyclic groups, having at least one heteroatom (O, S, or N) in at least one ring, preferably having one, two, or three heteroatoms selected from O, S, and N. Each ring of such a heteroatom-containing group may contain one or two oxygen or sulfur atoms or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and further provided that the ring contains at least one carbon atom. The nitrogen and sulfur atoms may optionally be oxidized, and the nitrogen atom may optionally be quaternized. Fused rings completing bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or fully unsaturated. Heterocyclic groups may be attached to any available nitrogen or carbon atom. As used herein, the terms “heterocycloalkyl”, “heterocyclo”, and “heterocyclic” include “heteroaryl” groups, as defined below.

[0057] In addition to the heteroaryl groups described below, exemplary monocyclic heterocyclic groups include azirrobutyl, pyrrolidinyl, oxacyclobutyl, imidazolinyl, oxazolinyl, isoxazolinyl, thiazolinyl, isothiazolinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiratinyl, 2-oxopiratinyl, 2-oxopiralinyl, 2-oxoazirroyl, azirroyl, 1-pyridoneyl, 4-piperidinoneyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1,3-dioxopentane, and tetrahydro-1,1-dioxothiopheneyl, etc. Exemplary bicyclic heterocyclic groups include quinoline cyclic groups.

[0058] The term "heteroaryl" refers to substituted and unsubstituted aromatic 5- or 6-membered monocyclic groups, 9- or 10-membered bicyclic groups, and 11- to 14-membered tricyclic groups, having at least one heteroatom (O, S, or N) in at least one ring, preferably having one, two, or three heteroatoms selected from O, S, and N. Each ring of a heteroaryl containing a heteroatom may contain one or two oxygen or sulfur atoms or one to four nitrogen atoms, provided that the total number of heteroatoms in each ring is four or less, and each ring has at least one carbon atom. Fused rings completing bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated, or unsaturated. Nitrogen and sulfur atoms may optionally be oxidized, and nitrogen atoms may optionally be quaternized. Heteroaryls as bicyclic or tricyclic groups must include at least one fully aromatic ring, but the other one or more fused rings may be aromatic or non-aromatic. Heteroaryls may be attached to any available nitrogen or carbon atom of any ring. Where the valence permits, if the other ring is a cycloalkyl or heterocyclic ring, it may optionally be substituted with =O (oxo).

[0059] As described herein, the term "pharmaceutically acceptable salt" means that the salt is not only physiologically acceptable to the subject, but also refers to a synthetic substance with pharmaceutical value, such as a salt formed as an intermediate during chiral resolution, although such intermediate salt cannot be directly given to the subject, but can play a role in obtaining the end product of the present invention.

[0060] As described herein, the term "disease" refers to a physical condition of the subject that is related to the disease described in this invention. Examples include peripheral arterial diseases and neurodegenerative diseases described in this invention.

[0061] The cancers covered by this invention include standard treatments such as surgery, radiation therapy, chemotherapy, and hormone therapy.

[0062] “Cancer” or “malignant tumor” refers to any of a variety of diseases characterized by uncontrolled, abnormal proliferation of cells, the ability of affected cells to spread locally or via the bloodstream and lymphatic system to other parts of the body (i.e., metastasis), and any of a number of characteristic structural and / or molecular features. “Cancer cells” refers to cells that have undergone early, intermediate, or late stages of multi-step tumor progression. Cancers include mesothelioma, neuroblastoma, rectal cancer, colon cancer, familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, stomach cancer, adenocarcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, kidney cancer, renal parenchymal carcinoma, ovarian cancer, cervical cancer, uterine cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, urinary tract cancer, melanoma, brain tumors, head and neck cancer, and acute lymphoblastic leukemia. ALL), chronic lymphocytic leukemia (CLL), acute myeloid leukemia (AML), chronic myeloid leukemia (CML), hepatocellular carcinoma, gallbladder cancer, bronchoma, advanced solid tumors, small cell lung cancer, metastatic non-small cell lung cancer, multiple myeloma, basal cell carcinoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myoma, liposarcoma, fibrosarcoma, Ewing sarcoma and plasmacytoma, lymphoma, pancreatic ductal adenocarcinoma, etc.

[0063] "Therapeutic effective amount" is an amount sufficient to provide therapeutic benefit in treating a disease, condition, or symptom, or sufficient to delay or minimize one or more symptoms associated with a disease, condition, or symptom. A therapeutically effective amount of a compound refers to the amount of a therapeutic agent (alone or in combination with other therapies) that provides therapeutic benefit in treating a disease, condition, or symptom. The term "therapeutic effective amount" may also encompass amounts that improve overall therapy, reduce or prevent symptoms or causes of a disease or symptom, or enhance the therapeutic efficacy of another therapeutic agent.

[0064] The compounds or pharmaceutical compositions containing them in this invention can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, intravenous drip, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0065] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including O / W, W / O, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0066] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0067] The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation. Detailed Implementation

[0068] The embodiments listed below help those skilled in the art to better understand the technical solutions of the present invention, but do not limit the present invention in any way.

[0069] For all the following embodiments, standard operations and methods known to those skilled in the art were used. Unless otherwise stated, all temperatures are expressed in °C (degrees Celsius). The structures of the compounds were determined by nuclear magnetic resonance spectroscopy (NMR) and / or mass spectrometry (MS).

[0070] The structures of the compounds of this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are expressed in parts per million (ppm). NMR was determined using a Bruker Avance-400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents and tetramethylsilane (TMS) as an internal standard.

[0071] Liquid chromatography-mass spectrometry (LC-MS) was performed using an ACQUITY UPLC ultra-high pressure liquid chromatograph for the liquid phase and a Xevo G2-S Qtof mass spectrometer for the mass spectrometry phase.

[0072] The starting materials used in the examples of this invention are known and commercially available, and can also be synthesized using or according to methods known in the art.

[0073] Example 1: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)piperidin-1-yl]-2-oxoacetamide (Compound 1)

[0074] Step 1: Synthesis of 1-benzyl-4-(4-bromo-2-nitrophenyl)-1,4-diaza-5-one

[0075] 1-Benzyl-1,4-diaza-5-one (10.0 g, 49.0 mmol, 1.0 equiv.), 4-bromo-1-iodo-2-nitrobenzene (17.7 g, 54.0 mmol, 1.1 equiv.), tris(dibenzylideneacetone)dipalladium (2.3 g, 2.5 mmol, 0.05 equiv.), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (2.8 g, 4.9 mmol, 0.1 equiv.), and potassium carbonate (20.0 g, 147.0 mmol, 3.0 equiv.) were dissolved in toluene (150 mL), purged with nitrogen three times, and refluxed at 100 °C for 18 hours. LC-MS and TLC monitoring were performed. After the reaction was complete, the reaction solution was concentrated directly, extracted with ethyl acetate and water, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain a pale yellow solid compound, 1-benzyl-4-(4-bromo-2-nitrophenyl)-1,4-diaza-5-one (13.5 g, yield 68.3%). LC-MS: [M+H]+=404.0593.

[0076] Step 2: Synthesis of 3-benzyl-8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0077] 1-Benzyl-4-(4-bromo-2-nitrophenyl)-1,4-diaza-5-one (13.5 g, 33.5 mmol, 1.0 Equiv.) and iron powder (9.4 g, 167.5 mmol, 5.0 Equiv.) were dissolved in acetic acid (140 mL) and refluxed at 100 °C for 1.5 h, monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated to remove acetic acid, and the pH was adjusted to alkaline with sodium carbonate by adding a small amount of water. The solution was extracted three times with ethyl acetate and water, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography (dichloromethane:methanol = 10:1) to obtain a yellow solid compound 3-benzyl-8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazoline (10.5 g, yield 88.3%). LC-MS:[M+H]+=356.0684.

[0078] Step 3: Synthesis of 3-benzyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0079] 3-Benzyl-8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (5.0 g, 14.1 mmol, 1.0 Equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (7.16 g, 28.2 mmol, 2.0 Equiv.), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (1.0 g, 1.41 mmol, 0.1 Equiv.), and potassium acetate (4.15 g, 42.3 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (100.0 mL), and the mixture was immediately purged with nitrogen three times. The mixture was stirred at 100 °C for 18 hours and monitored by LC-MS and TLC. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a white solid 3-benzyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (2.0 g, yield 35.2%). LCMS: [M+H]+ = 404.2561.

[0080] Step 4: Synthesis of (S)-6-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0081] 3-Benzyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (1 g, 2.48 mmol, 1.1 Equiv) and (S)-3-methyl-6-{[(trifluoromethyl)sulfonyl]oxy}-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (941 mg, 2.48 mmol, 1.1 Equiv) were prepared. 73 mmol, 1.1 Equiv; 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (181.5 mg, 0.248 mmol, 0.1 Equiv.) and sodium carbonate (788 mg, 7.44 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (10.0 mL) and water (3.0 mL). Nitrogen gas was immediately purged three times, and the mixture was stirred at 100 °C for 18 hours. The reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and purified by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a yellow oily compound (S)-6-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (1.1 g, yield 94%). LCMS: [M+H-Boc]+ = 473.2898.

[0082] Step 5: Synthesis of (S)-3-benzyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0083] (S)-6-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (1.1 g, 2.33 mmol, 1.0 Equiv.) was dissolved in dichloromethane (10.0 mL), followed by the addition of trifluoroacetic acid (3.98 g, 35 mmol, 15.0 Equiv.). The reaction was carried out at room temperature for 2 hours, and monitored by LCMS and TLC. After the reaction was complete, the pH was adjusted to alkaline with sodium carbonate, and the mixture was extracted three times with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain the compound (S)-3-benzyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (500 mg, yield 57.7%). LCMS: [M+H]+ = 373.2384.

[0084] Step 6: Synthesis of 3-benzyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0085] (S)-3-benzyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (500 mg, 1.34 mmol, 1.0 Equiv) was dissolved in methanol (10.0 mL). Sodium borohydride (127.3 mg, 3.35 mmol, 2.5 Equiv) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour and monitored by LC-MS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and purified by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a brown oily compound 3-benzyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (470 mg, yield 93.8%). LCMS: [M+H]+ = 375.2436.

[0086] Step 7: Synthesis of ethyl 2-[(2R,5S)-2-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate

[0087] 3-Benzyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (470 mg, 1.25 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (7.0 mL), and triethylamine (190 mg, 1.88 mmol, 1.5 Equiv.) was added. Under ice bath conditions, ethyl 2-chloro-2-oxoacetate (340 mg, 2.50 mmol, 2.0 Equiv.) was slowly added, and the reaction was stirred at room temperature for 1.5 hours. LCMS monitoring was performed. After the reaction was complete, the mixture was quenched with ethanol, concentrated, stirred, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain ethyl acetate 2-[(2R,5S)-2-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (300 mg, yield 50.6%). LCMS: [M+H]+ = 475.2709.

[0088] Step 8: Synthesis of ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate

[0089] Ethyl 2-[(2R,5S)-2-(3-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (300 mg, 0.63 mmol, 1.0 Equiv.) and palladium hydroxide on carbon (20%) (90 mg) were dissolved in methanol (6 mL), and then 2 drops of hydrochloric acid (6 mol / L) were added dropwise. Hydrogen gas was replaced three times, and the reaction was carried out at room temperature for 3 hours. The reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, mixed, and separated by column chromatography to obtain ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate (240 mg, yield 99%). LCMS: [M+H]+=385.2563.

[0090] Step 9: Synthesis of ethyl 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate

[0091] Ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate (50 mg, 0.26 mmol, 1.0 Equiv.), formaldehyde solution (2 mL), and 2 drops of acetic acid were added to methanol (4 mL). After reacting at room temperature for 0.5 hours, sodium borohydride acetate (110 mg, 0.52 mmol, 2.0 Equiv.) was added, and the reaction was carried out at room temperature for 1 hour. The reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and separated by column chromatography to obtain ethyl 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate (82 mg, yield 79%). LCMS: [M+H]+=399.2394.

[0092] Step 10: Synthesis of 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoacetic acid

[0093] Ethyl 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoethyl acetate (82 mg, 0.20 mmol, 1.0 Equiv.) and lithium hydroxide (9.6 mg, 0.40 mmol, 2.0 Equiv.) were added to tetrahydrofuran (2 mL) and water (2 mL), stirred at room temperature, and monitored by LCMS. After the reaction was completed, the pH was adjusted to acidic with dilute hydrochloric acid, and the product was concentrated to obtain crude 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoacetic acid (76 mg), which was directly used for the next reaction. LCMS: [M+H]+=371.2106.

[0094] Step 11: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)piperidin-1-yl]-2-oxoacetamide

[0095] 2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoacetic acid (76 mg, 0.20 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (3.0 mL), and 3-ethylpyridine-2,5-diamine (30.0 mg) was added. 0.22 mmol (1.1 Equiv.) and N,N-diisopropylethylamine (80.0 mg, 0.60 mmol, 3.0 Equiv.) were added and stirred at room temperature for 5 minutes. Then, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (157 mg, 0.40 mmol, 2.0 Equiv.) was added and stirred at room temperature for 1 hour. The reaction was monitored by LCMS. After the reaction was completed, the reaction solution was filtered and purified by high-performance liquid chromatography (HPLC) to obtain the purple powder compound N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(3-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)piperidin-1-yl]-2-oxoacetamide (18 mg, yield 18.4%). LCMS: [M+H]+=490.2932. 1 H NMR (400MHz, DMSO) δ10.40(s,1H),8.07(s,1H),7.50(s,3H),7.18(s,1H),5.59(d,J=87.2Hz,3H),4.29(d,J=8.8Hz,2H),3.15–3.07(m,2H),2. 67(dd,J=20.2,9.0Hz,5H),2.40(d,J=8.4Hz,5H),2.26(s,1H),2.13(s,1H),1.84(d,J=40.0Hz,2H),1.36(d,J=11.3Hz,1H),1.18–1.02(m,6H).

[0096] Example 2: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (compound 2)

[0097] Step 1: Synthesis of ethyl 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate

[0098] Ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)piperidin-1-yl]-2-oxoethyl acetate (50 mg, 0.130 mmol, 1.0 Equiv) was dissolved in N,N-dimethylformamide (1.5 mL), and bromoethane (56.8 mg, 0.521 mmol, 4.0 Equiv) and N,N-diisopropylethylamine (50.4 mg, 0.391 mmol, 3.0 Equiv) were added. The reaction was carried out at 50 °C for 18 hours, and the reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and purified by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain ethyl 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (64 mg). LCMS: [M+H]+ = 413.2559.

[0099] Step 2: Synthesis of 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid

[0100] Lithium hydroxide (30.8 mg, 1.286 mmol, 10.0 Equiv), methanol (2 mL), and water (0.2 mL) were added to ethyl acetate 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (64 mg, 0.128 mmol, 1.0 Equiv), and the mixture was reacted with LCMS for 1 hour at room temperature. After the reaction was complete, the solvent was evaporated, a small amount of water was added, the pH was adjusted to acidic with dilute hydrochloric acid, and the product was concentrated to obtain crude 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (49.0 mg), which was directly used in the next step of the reaction. LCMS: [M+H]+=385.2437.

[0101] Step 3: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide

[0102] 2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (49.0 mg, 0.128 mmol, 1.0 Equiv) was dissolved in N,N-dimethylformamide (2.0 mL), and 3-ethylpyridine-2,5-diamine (26.3 mg) was added. The mixture was stirred at room temperature for 5 minutes, and then 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (73.0 mg, 0.192 mmol, 1.5 Equiv) and N,N-diisopropylethylamine (41.3 mg, 0.320 mmol, 2.5 Equiv) were added. The mixture was then stirred at room temperature for 30 minutes, and the reaction was monitored by LCMS. After the reaction was completed, the reaction solution was filtered, and the solution was purified by high-performance liquid chromatography (HPLC) to obtain a light purple powder, N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(3-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (6.0 mg, yield 9.31%). LCMS: [M+H]+=504.3083. 1 H NMR(400MHz,MeOD)δ8.00(d,J=89.6Hz,1H),7.71–7.38(m,3H),7.30(s,1H),5 .62(d,J=169.4Hz,1H),4.36(s,2H),4.08–3.67(m,1H),3.42(d,J=14.1Hz,1H ),3.23(s,2H),2.86(d,J=10.3Hz,4H),2.71(q,J=7.3Hz,2H),2.48(d,J=23.4 Hz, 2H), 2.33 (s, 2H), 1.98 (d, J = 36.0Hz, 2H), 1.46 (s, 1H), 1.35–1.13 (m, 12H).

[0103] Example 3: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[3-ethyl-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (compound 3)

[0104] Step 1: Synthesis of tert-butyl 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-aza-1-carboxylic acid

[0105] 5-oxo-1,4-diaza-1-carboxylic acid tert-butyl ester (11.09 g, 51.82 mmol, 1.0 Equiv.), 4-bromo-1-iodo-2-nitrobenzene (18.7 g, 57 mmol, 1.1 Equiv.), tris(dibenzylacetone)dipalladium (2.37 mg, 2.59 mmol, 0.05 Equiv.), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (3.0 g, 5.18 mmol, 0.1 Equiv.), and potassium carbonate (21.45 g, 155.49 mmol, 3.0 Equiv.) were dissolved in toluene (200 mL), purged with nitrogen three times, and refluxed at 100 °C for 18 hours. LC-MS and TLC monitoring were performed. After the reaction was completed, the reaction solution was concentrated to remove toluene, extracted three times with ethyl acetate and water, and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, mixed, and separated by column chromatography to obtain 13 g of 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-aza-1-carboxylic acid tert-butyl ester. LC-MS: [M+H]+=414.2543. 1 H NMR (400MHz, DMSO-d6) δ8.18(d,J=2.3Hz,1H),8.01(dd,J=8.5,2.3Hz,1H),7.46(d,J=8.5 Hz, 1H), 3.91 (t, J = 4.5Hz, 2H), 3.64 (m, J = 66.7Hz, 4H), 2.68 (d, J = 7.1Hz, 2H), 1.42 (s, 9H).

[0106] Step 2: Synthesis of tert-butyl 4-(2-amino-4-bromophenyl)-5-oxo-1,4-aza-1-carboxylic acid

[0107] 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-aza-1-carboxylic acid tert-butyl ester (6.17 g, 14.94 mmol, 1.0 Equiv.), iron powder (4.18 g, 74.7 mmol, 5.0 Equiv.), and ammonium chloride (8.087 g, 149.4 mmol, 10 Equiv.) were dissolved in ethanol (120 mL) and water (30 mL) and refluxed at 80 °C for 2 hours. The reaction was monitored by LC-MS and TLC. After the reaction was complete, the reaction solution was filtered, the filtrate was concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1) to obtain 4-(2-amino-4-bromophenyl)-5-oxo-1,4-aza-1-carboxylic acid tert-butyl ester (4.41 g). LC-MS: [M+H]+ = 385.0818.

[0108] Step 3: Synthesis of tert-butyl 8-bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diaza-3-carboxylic acid

[0109] 4-(2-amino-4-bromophenyl)-5-oxo-1,4-aza-1-carboxylic acid tert-butyl ester (4.39 g, 11.43 mmol, 1.0 Equiv.) and ammonium chloride (8.087 g, 149.4 mmol, 10 Equiv.) were dissolved in ethanol (120 mL) and water (30 mL), and the mixture was refluxed at 80 °C for 5 hours. The reaction was monitored by LC-MS and TLC. After the reaction was complete, the reaction solution was concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1) to obtain 8-bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diaza-3-carboxylic acid tert-butyl ester (4.19 g). LC-MS: [M+H]+ = 310.0190.

[0110] Step 4: Synthesis of 8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0111] 8-Bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diaza-3-carboxylic acid tert-butyl ester (4.19 g, 11.45 mmol, 1.0 Equiv.) was dissolved in dichloromethane (100.0 mL), followed by the addition of trifluoroacetic acid (19.57 g, 171.7 mmol, 15.0 Equiv.). The reaction was carried out at room temperature for 2 hours, monitored by LCMS and TLC. After the reaction was completed, the pH was adjusted to alkaline with sodium carbonate, and the mixture was extracted with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain 8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-3-carboxylic acid (3.01 g). LCMS:[M+H]+=267.0334.

[0112] Step 5: Synthesis of 8-bromo-3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0113] 8-Bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (1 g, 3.76 mmol, 1.0 Equiv.), 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.61 g, 7.52 mmol, 2.0 Equiv.), and potassium carbonate (778 mg, 5.64 mmol, 1.5 Equiv.) were dissolved in acetonitrile (20 mL) and reacted at room temperature for 5 hours. The reaction was monitored by LCMS and TLC. After the reaction was complete, the reaction solution was concentrated, mixed, and separated by column chromatography to obtain a white solid 8-bromo-3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (690 mg). LCMS: [M+H]+=331.0437.

[0114] Step 6: Synthesis of 3-(2,2-difluoroethyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0115] 8-Bromo-3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (685 mg, 2.08 mmol, 1.0 Equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (1.056 g, 4.16 mmol, 2.0 Equiv.), 1,1-bis(diphenylphosphine)dipyridylferric palladium dichloride (152 mg, 0.208 mmol, 0.1 Equiv.), and potassium acetate (611 mg, 6.24 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (20.0 mL), and the mixture was immediately purged with nitrogen three times. The mixture was stirred at 100 °C for 17 hours and monitored by LC-MS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and subjected to column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a white solid 3-(2,2-difluoroethyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (690 mg). LCMS: [M+H]+ = 378.2282.

[0116] Step 7: Synthesis of (S)-6-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl]-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0117] 3-(2,2-difluoroethyl)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (690 g, 1.83 mmol, 1.1 Equiv), (S)-3-methyl-6-{[(trifluoromethyl)sulfonyl]oxy}-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (5 68 mg, 0.9 mmol, 0.9 Equiv; 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (134 mg, 0.183 mmol, 0.1 Equiv.) and sodium carbonate (582 mg, 5.49 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (12.0 mL) and water (4.0 mL). Nitrogen gas was immediately purged three times, and the mixture was stirred at 100 °C for 18 hours. The reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and subjected to column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a yellow oily crude product (S)-6-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl]-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl (800 mg). LCMS: [M+H-Boc]+ = 447.2553.

[0118] Step 8: Synthesis of (S)-3-(2,2-difluoroethyl)-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0119] (S)-6-(3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (800 g, 1.79 mmol, 1.0 Equiv.) was dissolved in dichloromethane (5.0 mL), followed by the addition of trifluoroacetic acid (3.06 g, 26.85 mmol, 15.0 Equiv.). The reaction was carried out at room temperature for 2 hours, and monitored by LCMS and TLC. After the reaction, the pH was adjusted to alkaline with sodium carbonate, and the mixture was extracted with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain (S)-3-(2,2-difluoroethyl)-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (287 mg). LCMS: [M+H]+ = 347.2041.

[0120] Step 9: Synthesis of 3-(2,2-difluoroethyl)-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine

[0121] (S)-3-(2,2-difluoroethyl)-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (287 mg, 0.83 mmol, 1.0 Equiv) was dissolved in methanol (5.0 mL). Sodium borohydride (78.8 mg, 2.1 mmol, 2.5 Equiv) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by LCMS and TLC. After the reaction was complete, the reaction solution was concentrated, mixed, and subjected to column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain 3-(2,2-difluoroethyl)-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (150 mg). LCMS: [M+H]+ = 349.2259.

[0122] Step 10: Synthesis of ethyl 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate

[0123] 3-(2,2-difluoroethyl)-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine (150 mg, 0.43 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (5.0 mL), and triethylamine (65 mg, 0.645 mmol, 1.5 Equiv.) was added. Under ice bath conditions, ethyl 2-chloro-2-oxoacetate (117 mg, 0.86 mmol, 2.0 Equiv.) was slowly added, and the reaction was stirred at room temperature for 1.5 hours. After the reaction was completed, the reaction was quenched with ethanol, concentrated, mixed, and separated by chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain ethyl acetate 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate (170 mg). LCMS: [M+H]+ = 449.2392.

[0124] Step 11: Synthesis of 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid

[0125] Ethyl 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diaza-8-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate (170 mg, 0.38 mmol, 1.0 Equiv.) and lithium hydroxide (91.2 mg, 3.8 mmol, 10.0 Equiv.) were dissolved in methanol (4 mL) and water (0.4 mL), and the reaction was carried out at room temperature for 0.5 hours. LCMS and TLC monitoring were performed. After the reaction was complete, the pH was adjusted to acidic with dilute hydrochloric acid, and the product was directly concentrated to obtain the crude product 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid, which was directly used in the next reaction. LCMS: [M+H]+=421.2070.

[0126] Step 12: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[3-ethyl-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide

[0127] 2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid (159.6 mg, 0.38 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (5.0 mL), and 3-ethylpyridine-2,5- Diamine (62.5 mg, 0.456 mmol, 1.2 Equiv.) and N,N-diisopropylethylamine (132 mg, 0.95 mmol, 2.5 Equiv.) were stirred at room temperature for 5 minutes, then 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (211.6 mg, 0.57 mmol, 1.5 Equiv.) was added, and the mixture was reacted at room temperature for 1 hour. After the reaction was completed, the reaction solution was filtered and the pink powder N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[3-ethyl-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (39 mg) was prepared and separated by LCMS monitoring. LCMS:[M+H]+=540.2900.1H NMR (400MHz, Methanol-d4) δ8.02(d,J=92.4Hz,1H),7.55(d,J=47.8Hz,3H),7.33(s,1H),6.23–5.33(m,2H),4.38(s,2H),4.01(s,1H),3.49(d,J =22.1Hz,1H),3.26(s,3H),3.11(d,J=19.4Hz,6H),2.51(d,J=23.3Hz,2 H), 2.36 (s, 2H), 1.97 (s, 2H), 1.57 (d, J = 53.2Hz, 1H), 1.41–1.15 (m, 8H).

[0128] Example 4: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (compound 11)

[0129] Step 1: Synthesis of tert-butyl 4-(4-chloro-2-nitrophenyl)-6-methyl-5-oxo-1,4-diazacyclohexane-1-carboxylate

[0130] First, tert-butyl 6-methyl-5-oxo-1,4-diazacyclohexane-1-carboxylate (4.0 g, 16.5 mmol, 1.0 equiv.), 1-bromo-4-chloro-2-nitrobenzene (3.90 g, 18.2 mmol, 1.1 equiv.), cesium carbonate (6.45 g, 19.8 mmol, 1.2 equiv.), tris(dibenzylacetone)palladium (1.51 g, 1.65 mmol, 0.1 equiv.), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (1.9 g, 3.3 mmol, 0.2 equiv.) were added to toluene (150 mL). The system was then purged with nitrogen three times, and the reaction was stirred at 100 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated to remove toluene, dissolved in ethyl acetate, filtered, and extracted with ethyl acetate and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain tert-butyl 4-(4-chloro-2-nitrophenyl)-6-methyl-5-oxo-1,4-diazacycloheptane-1-carboxylic acid (5.5 g). LCMS: [M-56+1] + =328.8.

[0131] Step 2: Synthesis of tert-butyl 8-chloro-5-methyl-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide-3-carboxylic acid

[0132] Compound 4-(4-chloro-2-nitrophenyl)-6-methyl-5-oxo-1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (5.5 g, 14.3 mmol, 1.0 equiv.), iron powder (3.2 g, 57.2 mmol, 4.0 equiv.), and ammonium chloride (6.12 g, 114.4 mmol, 8.0 equiv.) were dissolved in ethanol (50 mL) and water (20 mL). The system was then purged three times with nitrogen gas, and the mixture was stirred at 70 °C for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 8-chloro-5-methyl-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diazacyclo-3-carboxylic acid tert-butyl ester (2.44 g). LCMS: [M-56+1] + =298.1.

[0133] Step 3: Synthesis of 8-chloro-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0134] Compound 8-chloro-5-methyl-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene-3-carboxylic acid tert-butyl ester (2.44 g, 7.00 mmol, 1.0 equiv.) was dissolved in dioxane hydrochloride (30.0 mL) and stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to remove the solvent, diluted with water (6.0 mL), and the pH was adjusted to 8-9 with saturated sodium carbonate solution. The solution was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography to obtain compound 8-chloro-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene (1.45 g). LCMS: [M+H] + =236.1.

[0135] Step 4: Synthesis of 8-chloro-3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0136] Compound 8-chloro-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide (780 mg, 3.48 mmol, 1.0 equiv.), bromoethane (1.5 g, 13.14 mmol, 4.0 equiv.), and N,N-diisopropylethylamine (1.34 g, 10.44 mmol, 3.0 equiv.) were added to N,N-dimethylformamide (12 mL), and the mixture was stirred at 50 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated and subjected to column chromatography to obtain compound 8-chloro-3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide (280 mg). LCMS: [M+H] + =264.3.

[0137] Step 5: Synthesis of 3-ethyl-5-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0138] First, 8-chloro-3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (255 mg, 1.0 mmol, 1.0 equiv.) was dissolved in N,N-dimethylformamide (10 mL). Then, pinacol diboronate (762 mg, 3.0 mmol, 3.0 equiv.), potassium acetate (196 mg, 2.0 mmol, 2.0 equiv.), tridibenzylacetone dipalladium (137.4 mg, 0.15 mmol, 0.15 equiv.), and 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (143 mg, 0.30 mmol, 0.3 equiv.) were added. The system was purged with nitrogen three times, and the temperature was raised to 100 °C. The mixture was stirred for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and purified by column chromatography to obtain a pale yellow solid compound, 3-ethyl-5-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (200 mg). LCMS: [M+H] + =356.4.

[0139] Step 6: Synthesis of (3S)-6-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0140] Compound 3-ethyl-5-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoborhexacyclopentan-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (200 mg, 0.58 mmol, 1.0 equiv.) was dissolved in 1,4-dioxane (5 mL) and water (1.5 mL). (S) was added to the system. 3-Methyl-6-{[(trifluoromethyl)sulfonyl]oxy}-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (200 mg, 0.58 mmol, 1.0 equiv.), sodium carbonate (184 mg, 1.74 mmol, 3.0 equiv.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (42 mg, 0.058 mmol, 0.1 equiv.) were used. The system was purged with nitrogen three times, and the reaction was carried out at 90 °C for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and separated by column chromatography to obtain (3S)-6-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (100 mg). LCMS: [M+Na] + =425.4.

[0141] Step 7: Synthesis of 3-ethyl-5-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0142] The compound (3S)-6-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (100 mg, 0.24 mmol, 1.0 equiv.) was added to dichloromethane (4 mL), followed by the addition of trifluoroacetic acid (1 mL). The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to remove the solvent. The pH was adjusted to 8-9 with saturated sodium carbonate solution, and the solvent was evaporated under reduced pressure. Reversed-phase column chromatography yielded 3-ethyl-5-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazepine (56 mg). LCMS: [M+H] + =355.3.

[0143] Step 8: Synthesis of 3-ethyl-5-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0144] The obtained compound 3-ethyl-5-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (56 mg, 0.18 mmol, 1.0 equiv) was dissolved in methanol (3 mL). Sodium borohydride (13 mg, 0.36 mmol, 2.0 equiv) was added in portions at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 1 hour. After the reaction was complete, the reaction solution was concentrated and separated by column chromatography to obtain compound 3-ethyl-5-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (29 mg). LCMS: [M+H] + =327.4.

[0145] Step 9: Synthesis of ethyl 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate

[0146] The obtained compound 3-ethyl-5-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene (29 mg, 0.09 mmol, 1.0 equiv) and triethylamine (11 mg, 0.11 mmol, 1.2 equiv) were dissolved in N,N-dimethylformamide (6 mL), and then ethyl 2-chloro-2-oxoacetate (13.6 mg, 0.10 mmol, 1.1 equiv) was slowly added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated and separated by column chromatography to obtain ethyl 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (35 mg). LCMS: [M+H] + =427.3.

[0147] Step 10: Synthesis of compound 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid

[0148] Compound 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (35 mg, 0.09 mmol, 1.0 equiv) and lithium hydroxide (4 mg, 0.18 mmol, 2.0 equiv) were dissolved in tetrahydrofuran (1 mL) and water (1 mL), and reacted at room temperature for 1 hour. After the reaction was complete, the pH of the reaction solution was adjusted to 6.0–7.0 with dilute hydrochloric acid. The solvent was removed under reduced pressure to give the crude product 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (32.6 mg), which was used directly in the next reaction. LCMS: [M+H-COOH] + =355.3.

[0149] Step 11. Synthesis of compound N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazo-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetamide

[0150] 2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazo-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (32.6 mg, 0.09 mmol, 1.0 equiv.), 3-ethylpyridine-2,5-diamine (13.7 mg, 0.10 mmol, 1.2 equiv.), N,N-diisopropylethylamine (33.5 mg, 0.26 mmol, 3.0 equiv.), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (64.6 mg, 0.17 mmol, 2.0 equiv.) were added sequentially to N,N-dimethylformamide (1.5 mL), and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered to obtain a clear liquid, which was purified by preparative high-performance liquid chromatography (HPLC) to obtain a light purple-red solid N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(3-ethyl-5-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (16 mg). LCMS: [M+H] + =518.4.

[0151] Example 5: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetamide (compound 12)

[0152] Step 1: Synthesis of tert-butyl 4-(4-chloro-2-nitrophenyl)-3-methyl-5-oxo-1,4-diazacyclohexane-1-carboxylate

[0153] First, tert-butyl 3-methyl-5-oxo-1,4-diazacyclohexane-1-carboxylate (4.0 g, 16.5 mmol, 1.0 equiv.), 1-bromo-4-chloro-2-nitrobenzene (3.90 g, 18.2 mmol, 1.1 equiv.), cesium carbonate (6.45 g, 19.8 mmol, 1.2 equiv.), tris(dibenzylacetone)palladium (1.51 g, 1.65 mmol, 0.1 equiv.), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (1.9 g, 3.3 mmol, 0.2 equiv.) were added to toluene (150 mL). The system was then purged with nitrogen three times, and the reaction was stirred at 100 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated to remove toluene, dissolved in ethyl acetate, filtered, and extracted with ethyl acetate and water. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give tert-butyl 4-(4-chloro-2-nitrophenyl)-3-methyl-5-oxo-1,4-diazacycloheptane-1-carboxylic acid (5.6 g). LCMS: [M-56+1] + =328.8.

[0154] Step 2: Synthesis of tert-butyl 8-chloro-1-methyl-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide-3-carboxylic acid

[0155] Compound 4-(4-chloro-2-nitrophenyl)-3-methyl-5-oxo-1,4-diazacycloheptane-1-carboxylic acid tert-butyl ester (5.5 g, 14.3 mmol, 1.0 equiv.), iron powder (3.2 g, 57.2 mmol, 4.0 equiv.), and ammonium chloride (6.12 g, 114.4 mmol, 8.0 equiv.) were dissolved in a mixed solution of ethanol (50 mL) and water (20 mL). The system was purged with nitrogen three times, and the reaction was stirred at 70 °C for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to obtain compound 8-chloro-1-methyl-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazo[1,2-d][1,4]diazacyclo-3-carboxylic acid tert-butyl ester (2.51 g). LCMS:[M+H] + =336.1.

[0156] Step 3: Synthesis of 8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0157] Compound 8-chloro-1-methyl-1,2,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene-3-carboxylic acid tert-butyl ester (2.44 g, 7.00 mmol, 1.0 equiv.) was dissolved in dioxane hydrochloride (30.0 mL) and stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to remove the solvent, diluted with water (6.0 mL), and the pH was adjusted to 8-9 with saturated sodium carbonate solution. The solution was extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to obtain compound 8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene (1.35 g). LCMS: [M+H] + =236.1.

[0158] Step 4. Synthesis of compound 8-chloro-3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0159] Compounds 8-chloro-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide (780 mg, 3.48 mmol, 1.0 equiv.), bromoethane (1.5 g, 13.14 mmol, 4.0 equiv.), and N,N-diisopropylethylamine (1.34 g, 10.44 mmol, 3.0 equiv.) were added to N,N-dimethylformamide (12 mL). The mixture was stirred at 50 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated and purified by column chromatography to obtain compound 8-chloro-3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaporide (300 mg). LCMS: [M+H] + =264.1.

[0160] Step 5: Synthesis of 3-ethyl-1-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0161] First, 8-chloro-3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (255 mg, 1.0 mmol, 1.0 equiv.) was dissolved in N,N-dimethylformamide (10 mL). Then, pinacol diboronate (762 mg, 3.0 mmol, 3.0 equiv.), potassium acetate (196 mg, 2.0 mmol, 2.0 equiv.), tridibenzylacetone dipalladium (137.4 mg, 0.15 mmol, 0.15 equiv.), and 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl (143 mg, 0.30 mmol, 0.3 equiv.) were added. The system was purged with nitrogen three times, and the temperature was raised to 100 °C. The mixture was stirred for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and subjected to column chromatography to obtain a pale yellow solid compound, 3-ethyl-1-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazoline (218 mg). LCMS: [M+H] + =356.3.

[0162] Step 6: Synthesis of (3S)-6-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0163] 3-Ethyl-1-methyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (200 mg, 0.58 mmol, 1.0 equiv.) was dissolved in 1,4-dioxane (5 mL) and water (1.5 mL). (S)-3 1,4-Dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (200 mg, 0.58 mmol, 1.0 equiv.), sodium carbonate (184 mg, 1.74 mmol, 3.0 equiv.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (42 mg, 0.058 mmol, 0.1 equiv.) were used. The system was purged with nitrogen three times, and the reaction was carried out at 90 °C for 18 hours. After the reaction was complete, the reaction solution was filtered, concentrated, and separated by column chromatography to obtain (3S)-6-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (134 mg). LCMS: [M+H] + =425.3.

[0164] Step 7: Synthesis of 3-ethyl-1-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0165] The compound (3S)-6-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphen-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (100 mg, 0.24 mmol, 1.0 equiv.) was added to dichloromethane (4 mL), followed by the addition of trifluoroacetic acid (1 mL). The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to remove the solvent. The pH was adjusted to 8-9 with saturated sodium carbonate solution, and the solvent was evaporated under reduced pressure. Reversed-phase column chromatography yielded 3-ethyl-1-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazepine (65 mg). LCMS: [M+H] + =325.3.

[0166] Step 8: Synthesis of 3-ethyl-1-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide

[0167] The obtained compound 3-ethyl-5-methyl-8-[(S)-5-methyl-3,4,5,6-tetrahydropyridin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (56 mg, 0.18 mmol, 1.0 equiv) was dissolved in methanol (3 mL). Sodium borohydride (13 mg, 0.36 mmol, 2.0 equiv) was added in portions at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 1 hour. After the reaction was complete, the reaction solution was concentrated and separated by column chromatography to obtain compound 3-ethyl-1-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazolide (35 mg). LCMS: [M+H] + =327.4.

[0168] Step 9. Synthesis of compound 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazo-8-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate

[0169] The obtained compound 3-ethyl-1-methyl-8-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazaphene (29 mg, 0.09 mmol, 1.0 equiv) and triethylamine (11 mg, 0.11 mmol, 1.2 equiv) were dissolved in N,N-dimethylformamide (6 mL), and then ethyl 2-chloro-2-oxoacetate (13.6 mg, 0.10 mmol, 1.1 equiv) was slowly added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated and separated by column chromatography to obtain ethyl 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate (36 mg). LCMS: [M+H] + =427.3.

[0170] Step 10: Synthesis of 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid

[0171] Compound 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate (35 mg, 0.09 mmol, 1.0 equiv) and lithium hydroxide (4 mg, 0.18 mmol, 2.0 equiv) were dissolved in tetrahydrofuran (1 mL) and water (1 mL), and reacted at room temperature for 1 hour. After the reaction was complete, the pH of the reaction solution was adjusted to 6.0–7.0 with dilute hydrochloric acid. The solvent was removed under reduced pressure to give the crude product 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid (32.6 mg), which was used directly in the next reaction. LCMS: [M+H] + =385.5.

[0172] Step 11: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazo-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetamide

[0173] 2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazo-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid (32.6 mg, 0.09 mmol, 1.0 equiv.), 3-ethylpyridine-2,5-diamine (13.7 mg, 0.10 mmol, 1.2 equiv.), N,N-diisopropylethylamine (33.5 mg, 0.26 mmol, 3.0 equiv.), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (64.6 mg, 0.17 mmol, 2.0 equiv.) were added sequentially to N,N-dimethylformamide (1.5 mL), and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered to obtain a clear liquid, which was purified by preparative high-performance liquid chromatography (HPLC) to obtain the synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-(3-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl}-2-oxoacetamide (6.5 mg). LCMS: [M+H] + =518.7.

[0174] Example 6: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)piperidin-1-yl]-2-oxoacetamide (compound 13)

[0175] Step 1: Synthesis of 4-benzyl-1-(4-bromo-2-nitrophenyl)-1,4-diaza-2-one

[0176] First, compounds 4-benzyl-1,4-diaza-2-one (3.37 g, 16.5 mmol, 1.0 equiv.), 4-bromo-1-iodo-2-nitrobenzene (5.93 g, 18.2 mmol, 1.1 equiv.), potassium carbonate (6.45 g, 19.8 mmol, 1.2 equiv.), tris(dibenzylideneacetone)palladium (1.51 g, 1.65 mmol, 0.1 equiv.), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (1.9 g, 3.3 mmol, 0.2 equiv.) were added to toluene (150 mL). The system was then purged with nitrogen three times, and the reaction was stirred at 100 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated to remove toluene, dissolved in ethyl acetate, filtered, extracted with ethyl acetate and water, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give compound 4-benzyl-1-(4-bromo-2-nitrophenyl)-1,4-diaza-2-one (5.1 g). LCMS: [M-56+1] + =404.1.

[0177] Step 2: Synthesis of 2-benzyl-9-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0178] Compound 4-benzyl-1-(4-bromo-2-nitrophenyl)-1,4-diaza-2-one (5.1 g, 12.6 mmol, 1.0 equiv.), iron powder (2.8 g, 50.5 mmol, 4.0 equiv.), and ammonium chloride (5.4 g, 100.8 mmol, 8.0 equiv.) were dissolved in ethanol:water (50 mL: 20 mL). The system was then purged with nitrogen three times, and the mixture was stirred at 70 °C for 18 h. After the reaction was complete, the reaction solution was filtered, concentrated, extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give compound 2-benzyl-9-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (3.24 g, yield 72.4%). LCMS: [M+1] + =356.0.

[0179] Step 3: Synthesis of 2-benzyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0180] 2-Benzyl-9-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (3.06 g, 8.62 mmol, 1.0 Equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (3.28 g, 12.93 mmol, 1.5 Equiv.), 1,1-bis(diphenylphosphine)diberberine palladium dichloride (630.7 mg, 0.862 mmol, 0.1 Equiv.), and potassium acetate (2.53 g, 25.86 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (100.0 mL), and the mixture was immediately purged with nitrogen three times. The mixture was stirred at 100 °C for 17 hours. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain 2-benzyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (2.15 g). LCMS: [M+H] + =404.2.

[0181] Step 4: Synthesis of (S)-6-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0182] 2-Benzyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (1.34 g, 3.316 mmol, 1.0 Equiv) and 3-methyl-6-{[(trifluoromethyl)sulfonyl]oxy}-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (2.16 g, 3.3 mmol, 1.0 Equiv) were prepared. 16 mmol (1.0 Equiv), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (230 mg, 0.314 mmol, 0.1 Equiv.), and sodium carbonate (1.054 g, 9.948 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (30.0 mL) and water (10.0 mL). Nitrogen gas was immediately purged three times, and the mixture was stirred at 100 °C for 18 hours. The reaction was monitored by LC-MS and TLC. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain a yellow oily liquid (S)-6-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (1.05 g). LCMS:[M+H-Boc]+ =373.3.

[0183] Step 5: Synthesis of (S)-2-benzyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0184] (S)-6-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (944 mg, 2.0 mmol, 1.0 Equiv.) was dissolved in dichloromethane (10.0 mL), and then trifluoroacetic acid (3.42 g, 30.0 mmol, 15.0 Equiv.) was added. The reaction was carried out at room temperature for 2 hours, and monitored by LCMS and TLC. After the reaction was complete, the pH was adjusted to alkaline with saturated sodium carbonate, and the mixture was extracted with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain (S)-2-benzyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (425 mg). LCMS: [M+H] + =373.3.

[0185] Step 6: Synthesis of 2-benzyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0186] (S)-2-benzyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (416.6 mg, 1.12 mmol, 1.0 Equiv) was dissolved in methanol (10.0 mL), and sodium borohydride (106.4 mg, 2.8 mmol, 2.5 Equiv) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain a yellow solid 2-benzyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (345 mg). LCMS:[M+H] + =375.2.

[0187] Step 7: Synthesis of ethyl 2-[(2R,5S)-2-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate

[0188] 2-Benzyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (332.86 mg, 0.89 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (5.0 mL), and triethylamine (135 mg, 1.34 mmol, 1.5 Equiv.) was added. Under ice bath conditions, ethyl 2-chloro-2-oxoacetate (242 mg, 1.78 mmol, 2.0 Equiv.) was slowly added, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, the reaction was quenched with ethanol, and the reaction solution was concentrated. The product, ethyl 2-[(2R,5S)-2-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (345 mg), was obtained by column chromatography. LCMS: [M+H] + =475.3.

[0189] Step 8: Synthesis of ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate

[0190] Ethyl 2-[(2R,5S)-2-(2-benzyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (1.42 g, 3.0 mmol, 1.0 Equiv.) and palladium hydroxide on carbon (20%) (426 mg) were dissolved in methanol (25 mL), and then 2 drops of hydrochloric acid (6 mol / L) were added dropwise. Hydrogen gas was purged three times, and the reaction was carried out at room temperature for 18 hours. After the reaction was complete, the reaction solution was filtered, the filtrate was concentrated, and the crude product 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate (1.10 g) was obtained by column chromatography and used directly in the next step of the reaction. LCMS: [M+H] + =385.2.

[0191] Step 9: Synthesis of ethyl 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoethyl acetate

[0192] Ethyl 2-[(2R,5S)-5-methyl-2-(2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate (76.8 mg, 2.0 mmol, 1.0 equiv.) was added to methanol (2 mL), followed by dropwise addition of acetic acid (0.2 mL). The mixture was stirred at room temperature for 30 minutes, and then sodium borohydride acetate (847.6 mg, 4.0 mmol, 2.0 equiv.) was added. The mixture was then stirred at room temperature for 1 hour. After the reaction was complete, the reaction solution was concentrated to obtain the crude product ethyl 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoethyl acetate (65 mg), which was directly used in the next reaction. LCMS: [M+H] + =399.2.

[0193] Step 10: Synthesis of 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoacetic acid

[0194] Ethyl 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoethyl acetate (59.7 mg, 0.15 mmol, 1.0 equiv) and lithium hydroxide (7.2 mg, 0.3 mmol, 2.0 equiv) were dissolved in tetrahydrofuran (1 mL) and water (1 mL), and reacted at room temperature for 1 hour. After the reaction was complete, the pH of the reaction solution was adjusted to 6.0–7.0 with dilute hydrochloric acid. The solution was then concentrated to obtain the crude product 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoacetic acid (54 mg), which was directly used in the next reaction. LCMS: [M+H] + =371.2.

[0195] Step 11: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)piperidin-1-yl]-2-oxoacetamide

[0196] 2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)piperidin-1-yl]-2-oxoacetic acid (51.8 mg, 0.13 mmol, 1.0 equiv.), 3-ethylpyridine-2,5-diamine (17.81 mg, 0.13 mmol, 1.2 equiv.), N,N-diisopropylethylamine (50.31 mg, 0.39 mmol, 3.0 equiv.), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (98.8 mg, 0.26 mmol, 2.0 equiv.) were added sequentially to N,N-dimethylformamide (2.0 mL), and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered to obtain a clear liquid, which was then purified by preparative high-performance liquid chromatography (HPLC) to obtain N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-5-methyl-2-(2-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)piperidin-1-yl]-2-oxoacetamide (12.3 mg). LCMS: [M+H] + =490.3.

[0197] Example 7: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (compound 14)

[0198] Step 1: Synthesis of tert-butyl 9-bromo-4,5-dihydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-2(3H)-carboxylic acid

[0199] 4-(4-bromo-2-nitrophenyl)-3-oxo-1,4-diazacycloheptan-1-carboxylic acid tert-butyl ester (3.4 g, 8.232 mmol, 1.0 Equiv.), iron powder (1.84 g, 32.929 mmol, 4.0 Equiv.), and ammonium chloride (3.49 g, 65.859 mmol, 1.5 Equiv.) were dissolved in ethanol (25 mL) and water (10 mL), and the mixture was immediately refluxed at 70 °C for 18 hours. The reaction was monitored by LCMS and TLC. After the reaction was complete, the mixture was filtered, and the filtrate was mixed with a column chromatography solution (dichloromethane:methanol = 10:10.1% triethylamine) to give a yellowish-white solid 9-bromo-4,5-dihydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazacyclo-2(3H)-carboxylic acid tert-butyl ester (984 mg). LC-MS: [M-56+H] = 310.0193.

[0200] Step 2: Synthesis of 9-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0201] 9-Bromo-4,5-dihydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-2(3H)-carboxylic acid tert-butyl ester (984 mg, 2.556 mmol, 1.0 Equiv.) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (4.37 g, 38.338 mmol, 15.0 Equiv.) was added. The mixture was incubated at room temperature for 0.5 hours. The reaction was monitored by LCMS and TLC. After the reaction was complete, dichloromethane was evaporated to dryness, the pH was adjusted to alkaline with saturated sodium carbonate, the sample was mixed, and extracted with ethyl acetate and water by column chromatography. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain the crude product. After evaporation to a small amount of solvent, reverse-phase column chromatography was performed to obtain a white solid 9-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (530 mg). LC-MS: [M+H] = 266.0284.

[0202] Step 3: Synthesis of 9-bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0203] 9-Bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (530 mg, 2.00 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (5.0 mL), and bromoethane (872 mg, 8.00 mmol, 4.0 Equiv.) and N,N-diisopropylethylamine (774 mg, 6.00 mmol, 3.0 Equiv.) were added. The mixture was refluxed at 50 °C for 18 hours. The reaction was monitored by LCMS and TLC. After the reaction was complete, the sample was stirred and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain a yellow solid 9-bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (482 mg). LC-MS:[M+H]=294.0610.

[0204] Step 4: Synthesis of 2-ethyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0205] 9-Bromo-2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (482 mg, 1.645 mmol, 1.0 Equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (626.7 g, 2.468 mmol, 1.5 Equiv.), 1,1-bis(diphenylphosphine)dipyridylferric palladium dichloride (120.4 mg, 0.164 mmol, 0.1 Equiv.), and potassium acetate (483.6 mg, 4.935 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (20.0 mL), and the mixture was immediately purged with nitrogen three times. The mixture was stirred at 100 °C for 17 hours and monitored by LC-MS and TLC. After the reaction was complete, the sample was evaporated to dryness, stirred, and subjected to column chromatography (dichloromethane:methanol 10:1 + 0.1% triethylamine) to give a brown solid 2-ethyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxoboronacyclopentan-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (350 mg). LCMS: [M+H] = 342.2361.

[0206] Step 5: Synthesis of (S)-6-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0207] 2-Ethyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (350 mg, 1.026 mmol, 1.0 Equiv) and (S)-3-methyl-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (424.9 mg) 1,232 mmol, 1.2 Equiv; 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (75.0 mg, 0.103 mmol, 0.1 Equiv.) and sodium carbonate (326.4 mg, 3.079 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (15.0 mL) and water (5.0 mL). Nitrogen gas was immediately purged three times, and the reaction was stirred at 100 °C for 18 hours. The reaction was monitored by LCMS and TLC. After the reaction was complete, the sample was evaporated to dryness, stirred, and subjected to column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to give a black solid (S)-6-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (202.4 mg). LCMS: [M+H-Boc] = 311.2243.

[0208] Step 6: Synthesis of (S)-2-ethyl-9-(5-methyl-1,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0209] (S)-6-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (202.4 mg, 0.494 mmol, 1.0 Equiv.) was dissolved in dichloromethane (5.0 mL), followed by the addition of trifluoroacetic acid (844 mg, 7.405 mmol, 15.0 Equiv.). The reaction was carried out at room temperature for 1 hour, and monitored by LCMS and TLC. After the reaction was complete, the pH was adjusted to alkaline with sodium carbonate, the sample was mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain (S)-2-ethyl-9-(5-methyl-1,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (193 mg). LCMS: [M+H] = 311.2231.

[0210] Step 7: Synthesis of 2-ethyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide

[0211] (S)-2-ethyl-9-(5-methyl-1,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (193 mg, 0.623 mmol, 1.0 Equiv) was dissolved in methanol (8.0 mL), and sodium borohydride (59.2 mg, 1.558 mmol, 2.5 Equiv) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by LCMS and TLC. After the reaction was complete, the sample was evaporated to dryness, stirred, and subjected to column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to give the product 2-ethyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (65 mg). LCMS: [M+H] = 313.2459.

[0212] Step 8: Synthesis of ethyl 2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl)]-2-oxoethyl acetate

[0213] 2-Ethyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazolide (65 mg, 0.208 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (1.0 mL), and triethylamine (31.6 mg, 0.313 mmol, 1.5 Equiv.) was added. Under ice bath conditions, a solution of 2-chloro-2-oxoethyl acetate (56.7 mg, 0.417 mmol, 2.0 Equiv.) in N,N-dimethylformamide (1.0 mL) was slowly added, and the reaction was stirred at room temperature for 1 hour. After LCMS monitoring, the reaction was quenched with ethanol, concentrated, mixed, and separated by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain the product ethyl acetate 2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl)]-2-oxoethyl acetate (99.4 mg). LCMS: [M+H] = 413.2599.

[0214] Step 9: Synthesis of 2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid

[0215] Lithium hydroxide (57.8 mg, 2.407 mmol, 10.0 Equiv.), methanol (4 mL), and water (0.4 mL) were added to ethyl acetate 2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl)]-2-oxoethyl acetate (99.4 mg, 0.241 mmol, 1.0 Equiv.). The mixture was stirred at room temperature for 5 hours. The reaction was monitored by LCMS. After the reaction was complete, the pH was adjusted to acidic with dilute hydrochloric acid, and the solvent was evaporated to dryness before proceeding to the next step. LCMS: [M+H] = 385.2161.

[0216] Step 10: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide

[0217] 2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (92.5 mg, 0.241 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (2.5 mL), and 3-ethylpyridine-2,5-diamine was added. (49.5 mg, 0.362 mmol, 1.5 Equiv.) N,N-diisopropylethylamine (77.7 mg, 0.602 mmol, 2.5 Equiv.) was reacted with stirring at room temperature for 5 minutes, followed by the addition of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (137.4 mg, 0.362 mmol, 1.5 Equiv.), and the reaction was carried out at room temperature. After LCMS monitoring and reaction completion, the sample was filtered and sent to Prep-HPLC, then lyophilized to obtain a light purple powder, N-(6-amino-5-ethylpyridin-3-yl)-2-[(2R,5S)-2-(2-ethyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaphen-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (14 mg). LCMS: [M+H]=504.3083. 1H NMR (400MHz, MeOD) δ8.02(d,J=85.1Hz,1H),7.73-7.42(m,3H),7.34(s,1H),5.64(d,J=160.6Hz,1H),4.38(s,2H),4.19(s,2H),4.09-3.68( m,1H),3.46(d,J=31.2Hz,1H),3.24(s,2H),3.15(s,1H),2.50(s,4H),2.34(s,2H),1.97(s,4H),1.48(d,J=11.9Hz,1H),1.42-0.99(m,12H).

[0218] Example 8: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (compound 30)

[0219] Step 1: Synthesis of 4-benzyl-1-(4-bromo-2-nitrophenyl)-3-methyl-1,4-diazacyclopentan-2-one

[0220] First, 4-benzyl-3-methyl-1,4-diazacyclopentan-2-one (3.60 g, 16.5 mmol, 1.0 equiv.), 4-bromo-1-iodo-2-nitrobenzene (5.93 g, 18.2 mmol, 1.1 equiv.), potassium carbonate (6.45 g, 19.8 mmol, 1.2 equiv.), tris(dibenzylideneacetone)palladium (1.51 g, 1.65 mmol, 0.1 equiv.), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (1.9 g, 3.3 mmol, 0.2 equiv.) were added to toluene (150 mL). The system was then purged with nitrogen three times, and the reaction was stirred at 100 °C for 18 hours. After the reaction was complete, the reaction solution was concentrated to remove toluene, dissolved in ethyl acetate, filtered, extracted with ethyl acetate and water, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to give compound 4-benzyl-1-(4-bromo-2-nitrophenyl)-3-methyl-1,4-diazacyclopentan-2-one (5.3 g). LCMS: [M-56+1] + =418.1.

[0221] Step 2: Synthesis of 2-benzyl-9-bromo-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0222] Compound 4-benzyl-1-(4-bromo-2-nitrophenyl)-3-methyl-1,4-diaza-2-one (5.25 g, 12.6 mmol, 1.0 equiv.), iron powder (2.8 g, 50.5 mmol, 4.0 equiv.), and ammonium chloride (5.4 g, 100.8 mmol, 8.0 equiv.) were dissolved in ethanol:water (50 mL: 20 mL). The system was then purged with nitrogen three times, and the mixture was stirred at 70 °C for 18 h. After the reaction was complete, the reaction solution was filtered, concentrated, extracted with water and ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography to obtain compound 2-benzyl-9-bromo-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazoline (3.31 g). LCMS: [M+1] + =370.1.

[0223] Step 3: Synthesis of 2-benzyl-1-methyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0224] 2-Benzyl-9-bromo-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (3.18 g, 8.62 mmol, 1.0 Equiv.), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxoborane) (3.28 g, 12.93 mmol, 1.5 Equiv.), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (630.7 mg, 0.862 mmol, 0.1 Equiv.), and potassium acetate (2.53 g, 25.86 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (100.0 mL), and the mixture was immediately purged with nitrogen three times. The mixture was stirred at 100 °C for 17 hours. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain 1.8 g of 2-benzyl-1-methyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine. LCMS: [M+H] + =418.3.

[0225] Step 4: Synthesis of (S)-6-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester

[0226] 2-Benzyl-1-methyl-9-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (1.38 g, 3.316 mmol, 1.0 Equiv), 3-methyl-6-{[(trifluoromethyl)sulfonyl]oxy}-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (2.16 g, 3.316 mmol (1.0 Equiv), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (230 mg, 0.314 mmol, 0.1 Equiv.), and sodium carbonate (1.054 g, 9.948 mmol, 3.0 Equiv) were dissolved in 1,4-dioxane (30.0 mL) and water (10.0 mL). Nitrogen gas was immediately purged three times, and the mixture was stirred at 100 °C for 18 hours. The reaction was monitored by LC-MS and TLC. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain a yellow oily liquid (S)-6-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (1.13 g). LCMS:[M+H-Boc] + =387.3.

[0227] Step 5: Synthesis of (S)-2-benzyl-1-methyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0228] (S)-6-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (972 mg, 2.0 mmol, 1.0 Equiv.) was dissolved in dichloromethane (10.0 mL), and then trifluoroacetic acid (3.42 g, 30.0 mmol, 15.0 Equiv.) was added. The reaction was carried out at room temperature for 2 hours, and monitored by LCMS and TLC. After the reaction was complete, the pH was adjusted to alkaline with saturated sodium carbonate, and the mixture was extracted with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to yield (S)-2-benzyl-1-methyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (650.3 mg). LCMS: [M+H] + =387.3.

[0229] Step 6: Synthesis of 2-benzyl-1-methyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine

[0230] (S)-2-benzyl-1-methyl-9-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (432.32 mg, 1.12 mmol, 1.0 Equiv) was dissolved in methanol (10.0 mL), and sodium borohydride (106.4 mg, 2.8 mmol, 2.5 Equiv) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated and purified by column chromatography to obtain 2-benzyl-1-methyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (380 mg). LCMS:[M+H] + =389.3.

[0231] Step 7: Synthesis of ethyl 2-[(2R,5S)-2-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate

[0232] 2-Benzyl-1-methyl-9-[(2R,5S)-5-methylpiperidin-2-yl]-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazepine (345.32 mg, 0.89 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (5.0 mL), and triethylamine (135 mg, 1.34 mmol, 1.5 Equiv.) was added. Under ice bath conditions, ethyl 2-chloro-2-oxoacetate (242 mg, 1.78 mmol, 2.0 Equiv.) was slowly added, and the reaction was stirred at room temperature for 2 hours. After the reaction was completed, the reaction was quenched with ethanol, and the reaction solution was concentrated. The product, ethyl 2-[(2R,5S)-2-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (368 mg), was obtained by column chromatography. LCMS: [M+H] + =489.3.

[0233] Step 8: Synthesis of ethyl 2-[(2R,5S)-5-methyl-2-(1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate

[0234] Ethyl 2-[(2R,5S)-2-(2-benzyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl]-2-oxoethyl acetate (1.46 g, 3.0 mmol, 1.0 Equiv.) and palladium hydroxide on carbon (20%) (426 mg) were dissolved in methanol (25 mL), and then 2 drops of hydrochloric acid (6 mol / L) were added dropwise. Hydrogen gas was purged three times, and the reaction was carried out at room temperature for 18 hours. After the reaction was complete, the reaction solution was filtered, the filtrate was concentrated, and the crude product 2-[(2R,5S)-5-methyl-2-(1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate (1.25 g) was obtained by column chromatography and used directly in the next step of the reaction. LCMS: [M+H] + =399.2.

[0235] Step 9: Synthesis of ethyl 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate

[0236] Ethyl 2-[(2R,5S)-5-methyl-2-(1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate (76.8 mg, 2.0 mmol, 1.0 equiv.), 2,2,2-trifluoroethyltrifluoromethanesulfonate (1.61 g, 7.52 mmol, 2.0 equiv.), and potassium carbonate (778 mg, 5.64 mmol, 1.5 equiv.) were dissolved in acetonitrile (20 mL), and the mixture was stirred at room temperature for 5 hours. After the reaction was completed, the reaction solution was concentrated, mixed, and separated by column chromatography to obtain ethyl 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate (60 mg), LCMS: [M+H + =463.2.

[0237] Step 10: Synthesis of 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid

[0238] Ethyl 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoethyl acetate (59.7 mg, 0.15 mmol, 1.0 equiv) and lithium hydroxide (7.2 mg, 0.3 mmol, 2.0 equiv) were dissolved in tetrahydrofuran (1 mL) and water (1 mL), and reacted at room temperature for 1 hour. After the reaction was complete, the pH of the reaction solution was adjusted to 6.0–7.0 with dilute hydrochloric acid. The solution was then concentrated to obtain the crude product 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid (57.0 mg), which was directly used in the next reaction. LCMS: [M+H] + =435.2.

[0239] Step 11: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide

[0240] 2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetic acid (56.42 mg, 0.13 mmol, 1.0 equiv.), 3-ethylpyridine-2,5-diamine (17.81 mg, ... 0.13 mmol (1.2 equiv.), N,N-diisopropylethylamine (50.31 mg, 0.39 mmol, 3.0 equiv.) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (98.8 mg, 0.26 mmol, 2.0 equiv.) were added to N,N-dimethylformamide (2.0 mL), and the reaction was carried out at room temperature for 1 hour. After the reaction was complete, the reaction solution was filtered to obtain a clear liquid, which was then purified by preparative high-performance liquid chromatography (HPLC) to obtain N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-(2,2-difluoroethyl)-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl)-5-methylpiperidin-1-yl]-2-oxoacetamide (5.9 mg). LCMS: [M+H] + =554.3.

[0241] Example 9: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (compound 31)

[0242] Step 1: Synthesis of ethyl 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate

[0243] Ethyl 2-[(2R,5S)-5-methyl-2-(1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazazo-9-yl)piperidin-1-yl]-2-oxoethyl acetate (51.76 mg, 0.130 mmol, 1.0 Equiv) was dissolved in N,N-dimethylformamide (1.5 mL), and bromoethane (56.8 mg, 0.521 mmol, 4.0 Equiv) and N,N-diisopropylethylamine (50.4 mg, 0.391 mmol, 3.0 Equiv) were added. The reaction was carried out at 50 °C for 18 hours, and the reaction was monitored by LCMS and TLC. After the reaction was completed, the reaction solution was concentrated, mixed, and purified by column chromatography (dichloromethane:methanol = 10:1 + 0.1% triethylamine) to obtain ethyl acetate 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate (64 mg). LCMS: [M+H]+ = 427.3.

[0244] Step 2: Synthesis of 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid

[0245] Lithium hydroxide (30.8 mg, 1.286 mmol, 10.0 Equiv), methanol (2 mL), and water (0.2 mL) were added to ethyl acetate 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoethyl acetate (54.52 mg, 0.128 mmol, 1.0 Equiv), and the mixture was reacted with LCMS for 1 hour at room temperature. After the reaction was complete, the solvent was evaporated, a small amount of water was added, the pH was adjusted to acidic with dilute hydrochloric acid, and the product was concentrated to obtain crude 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid (45.0 mg), which was directly used in the next step of the reaction. LCMS: [M+H]+=399.2.

[0246] Step 3: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide

[0247] Dissolve 2-{(2R,5S)-2-(2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diaza-9-yl)-5-methylpiperidin-1-yl}-2-oxoacetic acid (45.0 mg, 0.128 mmol, 1.0 Equiv) in N,N-dimethylformamide (2.0 mL), and add 3-ethylpyridine-2,5-diamine (26 mg, 1.0 mL, 1.0 Equiv). 3 mg (0.192 mmol, 1.5 Equiv) and N,N-diisopropylethylamine (41.3 mg, 0.320 mmol, 2.5 Equiv) were stirred at room temperature for 5 minutes. Then, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (73.0 mg, 0.192 mmol, 1.5 Equiv) was added, and the mixture was reacted at room temperature for 30 minutes, monitored by LCMS. After the reaction was complete, the reaction solution was filtered to prepare and purified N-(6-amino-5-ethylpyridin-3-yl)-2-{(2R,5S)-2-[2-ethyl-1-methyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-a][1,4]diazaporin-9-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (6.0 mg). LCMS:[M+H]+=518.3.

[0248] Example 10: Synthesis of N-[6-amino-5-(trifluoromethyl)pyridin-3-yl]-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (Compound 38)

[0249] Step 1: Synthesis of 5-nitro-3-(trifluoromethyl)pyridine-2-amine

[0250] 2-Chloro-5-nitro-3-(trifluoromethyl)pyridine (800 mg, 3.5 mmol, 1.0 equiv.) and ammonia-methanol solution (8 mL, 7 M) were added to a sealed tube and reacted at room temperature for 16 hours. After the reaction was complete, the reaction solution was concentrated, diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 5-nitro-3-(trifluoromethyl)pyridine-2-amine (710 mg), which was directly used in the next reaction. LCMS: m / z = [M+H]208.0332.

[0251] Step 2: Synthesis of 3-(trifluoromethyl)pyridine-2,5-diamine

[0252] Iron powder (67 mg, 1.2 mmol, 5 equiv.) and ammonium chloride (130 mg, 2.4 mmol, 10 equiv.) were added to 4 mL of ethanol and 1 mL of water, and the mixture was reacted at 80 °C for 16 hours. After the reaction was complete, the reaction solution was filtered, the filtrate was concentrated, diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and separated by column chromatography to obtain 3-(trifluoromethyl)pyridine-2,5-diamine (30 mg). LCMS: m / z = [M+H] 177.0611.

[0253] Step 3: Synthesis of N-[6-amino-5-(trifluoromethyl)pyridin-3-yl]-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide

[0254] 2-[(2R,5S)-2-(3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (18 mg, 0.04 mmol, 1.0 equiv.) and 3-(trifluoromethyl)pyridine-2,5-diamine (12 mg, 0.06 mmol, 1.0 equiv.) were added. 1,N-diisopropylethylamine (14 mg, 0.11 mmol, 2.5 equiv.) and N,N-diisopropylethylamine (14 mg, 0.11 mmol, 2.5 equiv.) were dissolved in N,N-dimethylformamide (1.5 mL), stirred for 5 minutes, and then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (24 mg, 0.06 mmol, 1.5 equiv.) were added. The mixture was stirred at room temperature for 0.5 hours. After the reaction was completed, preparative chromatography purification yielded a white solid N-[6-amino-5-(trifluoromethyl)pyridin-3-yl]-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (4.4 mg). LCMS: m / z = [M+H] 580.2456. 1 H NMR (400MHz, DMSO-d6) δ10.89(d,J=37.2Hz,1H),8.42(d,J=31.7Hz,1H),8.08(d,J=3 8.0Hz,1H),7.56–7.48(m,2H),7.19(dd,J=25.1,8.7Hz,1H),6.76–6.00(m,3H),5.78 –5.21(m,1H),4.16(d,J=125.3Hz,3H),2.96(d,J=19.4Hz,9H),2.14(d,J=37.2Hz,1H ), 1.81 (d, J = 43.0Hz, 2H), 1.47 (d, J = 6.8Hz, 1H), 1.38 (s, 1H), 1.05 (d, J = 7.3Hz, 3H).

[0255] Example 11: Synthesis of N-(6-amino-5-cyclopropylpyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (compound 39)

[0256] Step 1: Synthesis of 3-cyclopropyl-5-nitropyridine-2-amine

[0257] 3-Bromo-5-nitropyridine-2-amine (900 mg, 4.1 mmol, 1.0 equiv.) was dissolved in 1,4-dioxane (10 mL) and water (2 mL). Cyclopropylboronic acid (710 mg, 8.2 mmol, 2.0 equiv.), potassium carbonate (1.1 g, 8.2 mmol, 2 equiv.), and [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (300 mg, 0.4 mmol, 0.1 equiv.) were added to the system. The system was purged with nitrogen three times, and the temperature was raised to 100 °C for 6 hours. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and separated by column chromatography to obtain 3-cyclopropyl-5-nitropyridine-2-amine (760 mg). LCMS: m / z = [M+H] 180.086.

[0258] Step 2: Synthesis of 3-cyclopropylpyridine-2,5-diamine

[0259] Pd (70 mg) was added to a methanol (5 mL) solution of compound 3-cyclopropyl-5-nitropyridine-2-amine (200 mg, 0.1 mmol, 1.0 equiv.), and hydrogen gas was purged three times. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the solution was filtered and separated by column chromatography to obtain 3-cyclopropylpyridine-2,5-diamine (110 mg). LCMS: m / z = [M+H] 150.0674.

[0260] Step 3: Synthesis of N-(6-amino-5-cyclopropylpyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide

[0261] 2-[(2R,5S)-2-(3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (18 mg, 0.04 mmol, 1.0 equiv.) and 3-cyclopropylpyridine-2,5-diamine (10 mg, 0.06 mmol, 1.0 equiv.) were added. 1.5 equiv.) and N,N-diisopropylethylamine (14 mg, 0.11 mmol, 2.5 equiv.) were dissolved in N,N-dimethylformamide (1.5 mL), stirred for 5 minutes, and then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (24 mg, 0.06 mmol, 1.5 equiv.) were added. The mixture was stirred for another 0.5 hours at room temperature. After the reaction was completed, the mixture was purified by preparative high-performance liquid chromatography (HPLC) to obtain a white solid N-(6-amino-5-cyclopropylpyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (2.3 mg). LCMS: m / z = [M+H] 552.2899. 1 H NMR (400MHz, DMSO-d6) δ10.29–10.01(m,1H),7.57–7.45(m,2H),7.28–7.19(m,1H),7.14(d,J =9.0Hz,1H),6.31(d,J=13.2Hz,1H),6.26–6.16(m,1H),5.93–5.21(m,2H),4.31(d,J=7.3Hz,2 H),3.48(d,J=13.1Hz,1H),3.30–3.22(m,1H),3.18–2.88(m,10H),2.30(d,J=18.8Hz,1H),2. 20 (s, 2H), 2.02 (s, 2H), 1.81 (d, J = 47.5Hz, 2H), 1.38 (t, J = 15.3Hz, 1H), 1.07 (t, J = 9.2Hz, 3H).

[0262] Example 12: Synthesis of N-(6-amino-5-methoxypyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (compound 40)

[0263] Step 1: Synthesis of 3-methoxy-5-nitropyridine-2-amine

[0264] In a sealed tube, 8 mL of ammonia-methanol solution (7 M) was added to compound 2-chloro-3-methoxy-5-nitropyridine (800 mg, 4 mmol, 1.0 equiv.), and the reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the solvent was evaporated, diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride, and the organic phases were combined, dried over anhydrous sodium sulfate, and filtered to give 3-methoxy-5-nitropyridine-2-amine (400 mg). LCMS: m / z = [M+H] 170.0156.

[0265] Step 2: Synthesis of 3-methoxypyridine-2,5-diamine

[0266] Pd (70 mg) was added to a methanol (5 mL) solution of 3-methoxy-5-nitropyridine-2-amine (200 mg, 1 mmol, 1.0 equiv.), and hydrogen gas was purged three times. The reaction was allowed to proceed overnight at room temperature. After the reaction was complete, the mixture was filtered and separated by column chromatography to obtain 100 mg of 3-methoxypyridine-2,5-diamine. LCMS: m / z = [M+H] 140.0413.

[0267] Step 3: Synthesis of N-(6-amino-5-methoxypyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide

[0268] 2-[(2R,5S)-2-(3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl)-5-methylpiperidin-1-yl]-2-oxoacetic acid (20 mg, 0.05 mmol, 1.0 equiv.) and 3-methoxypyridine-2,5-diamine (10 mg, 0.07 mmol) were added. N,N-diisopropylethylamine (15 mg, 0.12 mmol, 2.5 equiv.) and N,N-diisopropylethylamine (15 mg, 0.12 mmol, 2.5 equiv.) were dissolved in N,N-dimethylformamide (1.5 mL), stirred for 5 minutes, and then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (27 mg, 0.07 mmol, 1.5 equiv.) was added. The mixture was stirred at room temperature for 0.5 hours. After the reaction was complete, the mixture was purified by preparative high-performance liquid chromatography to obtain a white solid N-(6-amino-5-methoxypyridin-3-yl)-2-{(2R,5S)-2-[3-(2,2-difluoroethyl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazo[1,2-d][1,4]diazazo-8-yl]-5-methylpiperidin-1-yl}-2-oxoacetamide (9 mg). LCMS:m / z[M+H]+:542.2692. 1 H NMR (400MHz, DMSO-d6) δ10.36(d,J=133.7Hz,1H),8.13–7.74(m,1H),7.61–7.23(m,3H),5.60(d,J=59.7Hz,3H),4.92(s,2H),4 .28–3.46(m,6H),2.44–2.28(m,2H),2.12(s,1H),2.05–1.79(m,3H),1.23(s,1H),1.12(t,J=7.4Hz,3H),1.01(d,J=6.8Hz,3H).

[0269] Example 13: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide (compound 10)

[0270] Step 1: Synthesis of tert-butyl 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-diazane-1-carboxylic acid ester

[0271] To compound tert-butyl 5-oxo-1,4-diazane-1-carboxylic acid ester (19.98 g, 0.09 mol, 1.0 equiv.), 4-bromo-1-iodo-2-nitrobenzene (33.7 g, 0.1 mol, 1.1 equiv.), potassium carbonate (38.7 g, 0.28 mol, 3.0 equiv.), tris(dibenzylideneacetone)dipalladium (4.3 g, 0.0045 mol, 0.05 equiv.), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (5.4 g, 0.01 mol, 0.1 equiv.), and toluene (300 mL) were added. The mixture was immediately purged with nitrogen three times and refluxed at 100 °C for 18 hours. After the reaction was complete, toluene was evaporated, and the mixture was extracted with ethyl acetate and water. The organic phase was dried, concentrated, and separated by column chromatography (DCM: EA, EA% = 5.7%) to obtain 7.9 g of tert-butyl 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-diazane-1-carboxylic acid ester. The impurities were slurried to give 18 g of yellow solid tert-butyl 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-diazane-1-carboxylic acid ester (yield 69%), LCMS: [M+H-Boc] = 314.01.

[0272] Step 2: Synthesis of tert-butyl-8-bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazolium[1,2-D][1,4]diazepine-3-carboxylic acid ester

[0273] Compound tert-butyl 4-(4-bromo-2-nitrophenyl)-5-oxo-1,4-diazane-1-carboxylic acid ester (80 g, 0.194 mol, 1.0 equiv.), iron powder (54.2 g, 0.968 mol, 5.0 equiv.), and ammonium chloride (156.9 g, 2.91 mol, 15.0 equiv.) were dissolved in ethanol (800 mL) and water (200 mL) and refluxed at 80 °C overnight. After the reaction was complete, the mixture was filtered while hot, the filtrate was evaporated to dryness with ethanol, extracted with ethyl acetate, the organic phase was evaporated to dryness, and slurryed to give 50 g of light brown solid tert-butyl 8-bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazolium[1,2-D][1,4]diazane-3-carboxylic acid ester. LC-MS: [M+H-56] = 310.02.

[0274] Step 3: Synthesis of 8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine

[0275] 11 g (30 mmol, 1.0 equiv.) of tert-butyl-8-bromo-1,2,4,5-tetrahydro-3H-benzo[4,5]imidazole[1,2-D][1,4]diazepine-3-carboxylic acid ester was dissolved in trifluoroacetic acid (30 ml) and dichloromethane (30 ml) and stirred at room temperature for 1 hour. The solvent was distilled off under reduced pressure, diluted with water, and the pH was adjusted to 8-9 with saturated sodium carbonate solution. The mixture was extracted with ethyl acetate, washed with saturated sodium chloride, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was distilled off under reduced pressure. The mixture was then separated by reversed-phase column chromatography to obtain 4.2 g of colorless oily 8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazole[1,2-d][1,4]diazepine. LC-MS: [M+H] = 266.09.

[0276] Step 4: Synthesis of 8-bromo-3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine

[0277] Compound 8-bromo-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine (1.32 g, 4.981 mmol, 1.0 equiv.) was dissolved in methanol (40 mL). (1-methoxycyclopropoxy)trimethylsilane (4.33 g, 24.906 mmol, 5.0 equiv.) and acetic acid (20 drops) were added to the system. After stirring at room temperature for 30 minutes, sodium cyanoborohydride (938.4 mg, 14.943 mmol, 3.0 equiv.) was added, and the reaction was carried out at 50 °C for 12 h. After the reaction was complete, column chromatography (D:M = 10:1 + triethylamine) yielded 1.88 g of a yellow oily liquid, 8-bromo-3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazole[1,2-d][1,4]diazepine, with LC-MS concentrations of [M+H] = 306.06. NMR (400MHz, DMSO) δ7.71(d,J=2.0Hz,1H),7.50(d,J=8.5Hz,1H),7.33(dd,J=8.6,1.9Hz,1H),4.32–4.25(m,2H ),3.16–3.08(m,2H),2.94–2.83(m,4H),2.03(dq,J=6.4,3.3Hz,1H),0.56–0.49(m,2H),0.44(q,J=2.9Hz,2H).

[0278] Step 5: Synthesis of 3-cyclopropyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine

[0279] Add 4,4,4',4',5',5'-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine (1.99 g, 6.524 mmol, 1.0 equiv.), 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (477.6 mg, 0.652 mmol, 0.1 equiv.), and potassium acetate (1.92 g, 19.574 mmol, 3.0 equiv.) to 8-bromo-3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine (4.99 g, 6.524 mmol, 1.0 equiv.), and dissolve 1,4-dioxane (40.0 mL). Immediately purge with nitrogen three times and stir at 100 °C for 18 hours. After the reaction was completed, the sample was dried by rotary evaporation, mixed, and subjected to column chromatography (PE:EA = 1:1) to obtain 1.29 g of brown solid 3-cyclopropyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine, LC-MS: [M+H] = 354.23.

[0280] Step 6: Synthesis of tert-butyl(S)-6-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazepine-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid ester

[0281] The compound 3-cyclopropyl-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboron-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine (1.33 g, 3.768 mmol, 1.0 equiv.) (S)-3-methyl-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester 1,1-bis(diphenylphosphine)dimerferropalladium dichloride (275.8 mg, 0.377 mmol, 0.1 Equiv.) and sodium carbonate (1.2 g, 11.303 mmol, 3.0 Equiv.) were dissolved in 1,4-dioxane (30.0 mL) and water (10.0 mL), and nitrogen gas was immediately purged three times. The mixture was stirred at 90 °C for 18 hours. After the reaction was complete, the sample was dried by rotary evaporation, mixed, and subjected to column chromatography (PE:EA = 1:1) to obtain 1.38 g of yellow oily liquid tert-butyl(S)-6-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazepine-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid ester. LC-MS: [M+H-Boc] = 323.23.

[0282] Step 7: Synthesis of (S)-3-cyclopropyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine

[0283] The compound tert-butyl(S)-6-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazo-8-yl)-3-methyl-3,4-dihydropyridine-1(2H)-carboxylic acid ester (1.38 g, 3.270 mmol, 1.0 Equiv.) was dissolved in dichloromethane (10.0 mL), and then trifluoroacetic acid (5.6 g, 49.052 mmol, 15.0 Equiv.) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was completed, the pH was adjusted to alkaline with sodium carbonate, the sample was mixed, and separated by column chromatography (D:M = 10:1 + 0.1% triethylamine) to obtain 2.0 g (containing salt) of yellow oily liquid (S)-3-cyclopropyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine, LC-MS: [M+H] = 323.23.

[0284] Step 8: Synthesis of 3-cyclopropyl-8-((2R,5S)-5-methylpiperidin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine

[0285] The compound (S)-3-cyclopropyl-8-(5-methyl-3,4,5,6-tetrahydropyridin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine (1.0 g, 3.106 mmol, 1.0 Equiv.) was dissolved in methanol (10.0 mL), and sodium borohydride (310.6 mg, 8.175 mmol, 2.5 Equiv.) was added in portions at 0 °C. The mixture was stirred at room temperature for 1 hour. After the reaction was complete, the sample was dried by rotary evaporation, mixed, and subjected to column chromatography (D:M = 10:1 + 0.1% triethylamine) to obtain a yellow oily liquid 3-cyclopropyl-8-((2R,5S)-5-methylpiperidin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine 1.10 g, LC-MS: [M+H] = 325.24.

[0286] Step 9: Synthesis of ethyl acetate 2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxyethyl acetate

[0287] Compound 3-cyclopropyl-8-((2R,5S)-5-methylpiperidin-2-yl)-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazolium[1,2-d][1,4]diazepine (1.10 mg, 3.086 mmol, 1.0 Equiv.) was dissolved in N,N-dimethylformamide (3.0 mL), and triethylamine (467.6 mg, 4.629 mmol, 1.5 Equiv.) was added. Under ice bath conditions, a solution of ethyl 2-chloro-2-oxoacetate (839.5 mg, 6.173 mmol, 2.0 Equiv.) in N,N-dimethylformamide (1.0 mL) was slowly added, and the reaction was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was quenched with ethanol, concentrated, mixed, and separated by column chromatography (D:M = 10:1 + 0.1% triethylamine) to obtain 375 mg of ethyl acetate 2-((2R, 5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxyethyl acetate. LC-MS: [M+H] = 425.26.

[0288] Step 10: Synthesis of 2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxyacetic acid

[0289] Compound 2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxyethyl acetate (375 mg, 0.884 mmol, 1.0 Equiv.) was mixed with lithium hydroxide (212.3 mg, 8.844 mmol, 10.0 Equiv.), methanol (10 mL), and water (1 mL), and stirred at room temperature for 1 hour. After the reaction was complete, the pH was adjusted to acidic with dilute hydrochloric acid, and the solvent was evaporated to dryness before proceeding to the next step. LCMS: [M+H] = 397.22.

[0290] Step 11: Synthesis of tert-butyl (5-(2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamido)-3-ethylpyridin-2-yl)carbamate

[0291] 2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxyacetic acid (350.1 mg, 0.884 mmol, 1.0 Equiv) was dissolved in N,N-dimethylformamide (3 mL), and tert-butyl(5-amino-3-ethylpyridin-2-yl)aminomethylformamide was added. The ester (314.2 mg, 1.326 mmol, 1.5 Equiv.) and N,N-diisopropylethylamine (307.2 mg, 2.210 mmol, 2.5 Equiv.) were stirred at room temperature for 5 minutes, and then 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (503.9 mg, 1.326 mmol, 1.5 Equiv.) was added, and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, reversed-phase column chromatography yielded 349 mg of white solid tert-butyl (5-(2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamido)-3-ethylpyridin-2-yl)carbamate, LCMS: [M+H-Boc]=516.31.

[0292] Step 12: Synthesis of N-(6-amino-5-ethylpyridin-3-yl)-2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide

[0293] The compound tert-butyl (5-(2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-D][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamido)-3-ethylpyridin-2-yl)carbamate (349 mg, 0.567 mmol, 1.0 Equiv.) was dissolved in dichloromethane (5.0 mL), and then trifluoroacetic acid (970.4 mg, 8.512 mmol, 15.0 Equiv.) was added. The reaction was carried out at room temperature for 1 hour. After the reaction was complete, the solution was evaporated to dryness, dissolved in methanol (1.5 mL), filtered, and sent to Prep-HPLC. Lyophilization yielded 46 mg of N-(6-amino-5-ethylpyridin-3-yl)-2-((2R,5S)-2-(3-cyclopropyl-2,3,4,5-tetrahydro-1H-benzo[4,5]imidazol[1,2-d][1,4]diazepine-8-yl)-5-methylpiperidin-1-yl)-2-oxoacetamide. LCMS: [M+H] = 516.31, 1H NMR (400MHz, DMSO) δ10.58(d,J=17.3Hz,1H),8.07(d,J=29.8Hz,1H),7.53(d,J=20.9Hz, 3H),7.23(s,1H),5.83–5.19(m,3H),4.32(s,2H),3.76(dd,J=226.3,13.5Hz,1H),3.29(s ,1H),3.12(s,2H),2.94(d,J=9.3Hz,4H),2.49–2.27(m,3H),2.09(d,J=7.3Hz,2H),1.82( d,J=33.7Hz,2H),1.46–1.30(m,1H),1.12(dd,J=39.0,6.9Hz,6H),0.47(d,J=3.0Hz,4H).

[0294] The compounds listed in Table 1 below were prepared using methods similar to those described in the examples, with appropriate variations in the amounts of reactants and reagents, protection and deprotection, solvents, and reaction conditions. Characterization data for the compounds are summarized in Table 1 below.

[0295] Table 1: Structure and characterization of some compounds

[0296] Experimental Example 1: The effect of the compound of the present invention on HCT116 MTAP - / - Assay for cell proliferation

[0297] Experimental Objective: The purpose of this test case is to test the effect of the compound on HCT116 MTAP.- / - Cell proliferation.

[0298] Background and Principle: Protein arginine methyltransferase 5 (PRMT5) can methylate various proteins and plays an important role in biological processes such as gene expression, splicing, and DNA damage repair. Methionyl adenosine phosphorylase (MTAP) often exhibits co-deletion with the commonly used tumor suppressor gene CDKN2A, with this co-deletion rate reaching 9%–15% in tumors. Studies have found that inhibiting PRMT5 has a synthetic lethal effect in MTAP-deficient tumors. Therefore, this study investigated the effect of compounds on HCT116MTAP... - / - The inhibition rate of cell proliferation can be used to screen PRMT5 protein inhibitors.

[0299] Specific experimental procedure:

[0300] HCT116 MTAP knockout cells were constructed and single clones were screened. HCT116 MTAP cells in logarithmic growth phase were then... - / - WT cells were seeded in 96-well plates at 90 μL per well, 1000 cells / well, and incubated overnight at 37°C. The next day, 10 μL of different concentrations of the compound were added, with a maximum concentration of 100 μM, diluted 10-fold in 9 gradients (DMSO final concentration 1%), and incubated at 37°C for 7 days. On day 7, the old medium was aspirated, and 110 μL of medium (medium to CCK8 ratio 100:10) was added, and incubated at 37°C for 1–4 hours. Absorbance was measured at 450 nM, and IC50 was calculated using GraphPad software. Compounds were screened by comparison with positive control drugs.

[0301] IC50 (half maximal inhibitory concentration) refers to the half-maximal inhibitory concentration of a measured antagonist. It indicates the amount by which a drug or substance (inhibitor) inhibits a certain biological process (or certain substances contained in this process, such as enzymes, cell receptors, or microorganisms). The PRMT5 inhibitor TNG908 was used as a positive reference compound, and its preparation method is based on the examples in patent document WO2022026892. Its structure is: The measurement results are shown in Table 2 below:

[0302] Table 2: IC50 Measurement Results

[0303] Test results show that the representative compound of this invention has a good inhibitory effect on HCT116 MTAP- / - cells, which is better than the positive control, while the inhibitory effect on HCT116 WT cells is weak, showing excellent selectivity.

[0304] Experimental Example 2: Test of the inhibitory effect of the compounds of the present invention on PRMT5 enzyme function

[0305] Experimental objective: To detect the inhibitory effect of compounds on PRMT5 enzyme function using the MTase-Glo method.

[0306] Background and Principle: The gene responsible for the synthetic lethality of PRMT5 is MTAP (methionine phosphorylase). MTAP participates in the metabolism of 2-methylthioadenosine (MTA) and the regeneration of methionine required for the synthesis of SAM. The deletion of the MTAP gene leads to the accumulation of MTA in the cell, which competes with the PRMT5 substrate S-adenosyl-L-methionine (SAM), resulting in decreased PRMT5 activity. Based on the MTase-Glo method, this study utilizes the property of methyltransferase (PRMT5) to donate a methyl group to the substrate SAM (S-adenosylmethionine) to generate SAH. PRMT5 methyltransferase activity assays are performed in the presence and absence of MTA to screen for selective PRMT5 inhibitors.

[0307] Specific experimental procedure: The conversion of s-adenosylmethionine (SAM) to s-adenosylhomocysteine ​​(SAH) by the recombinant PRMT5:MEP50 enzyme complex was monitored using the MTase-Glo methyltransferase fluorescence method in the presence or absence of 5'-methylthioadenosine (MTA). The enzymatic reaction was carried out in white 96-well plates. The reaction buffer contained 20 mM bicine (pH 7.60), 25 mM NaCl, 1 mM DTT, and 0.1% (w / v) CHAPS. The test compound and positive control were prepared by 10-fold dilution, 9 concentration gradients, 3 replicates, with a maximum concentration of 100 μM. In the absence of MTA, the compound was incubated with a reaction mixture containing 1 nM recombinant PRMT5:MEP50 enzyme complex, 2.5 μM H41-21 histone peptide, and 2.5 μM s-adenosylmethionine (SAM). Similarly, in the presence of MTA, the compound was incubated with a mixture of 2 nM enzyme complex, 1.5 μM MTA, 2.5 μM H41-21 histone peptide, and 2.5 μM SAM. After incubation at room temperature for 5 h, TFA was added to each well to terminate the reaction. Subsequently, 5X MTase-Glo™ reagent was added to each well, and the mixture was incubated at room temperature to convert SAH generated in the reaction to ADP. After 10 min, MTase-Glo assay solution was added to each well, and the mixture was incubated at room temperature for 30 min before detection using a microplate reader.

[0308] Experimental Example 3: In vivo pharmacokinetic test of the compound of the present invention in rats

[0309] Experimental Objective: Using SD rats as the test substance, the plasma concentration of the drug at different time points after gavage administration was determined by LC-MS / MS. The pharmacokinetic behavior of the compound of this invention in rats was investigated, and its pharmacokinetic characteristics were evaluated.

[0310] Experimental procedure:

[0311] Each group consisted of 3 healthy male and female SD rats aged 6-8 weeks.

[0312] The compound was prepared on the day of gavage administration, using a solvent of 10% DMSO + 15% Solutol HS-15 and 0.5% sodium carboxymethyl cellulose aqueous solution (75%).

[0313] SD rats were fasted overnight before administration and fed 4 hours after administration.

[0314] Rats were administered the drug by gavage. Blood samples were collected via jugular vein at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h post-administration. Approximately 0.2 mL of blood was collected per sample. The samples were anticoagulated with K2-EDTA and placed on ice after collection. Plasma samples were separated by centrifugation within 1 hour (centrifugation conditions: 6800 g, 6 minutes, 2-8℃). Plasma samples were stored at -80℃ before analysis.

[0315] Samples were analyzed by LC-MS / MS, and the intraday accuracy of quality control samples was evaluated, with over 66.7% of the quality control samples required to have an accuracy between 80-120%. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on blood drug concentration data at different time points, providing AUC0-t / D (converted to per mg / kg) and Cmax / D (converted to per mg / kg).

[0316] Table 2. Pharmacokinetic results of the compounds of this invention in SD rats.

[0317] Conclusion: Some compounds of this invention exhibit superior pharmacokinetic characteristics compared to the positive control compounds.

[0318] Experimental Example 4: Pharmacokinetic Test of the Compounds of the Invention in Mice

[0319] Experimental Objective: Using ICR rats as the test substance, the plasma concentration of the drug at different time points after gavage administration was determined by LC-MS / MS. The pharmacokinetic behavior of the compound of this invention in mice was investigated, and its pharmacokinetic characteristics were evaluated.

[0320] Experimental procedure:

[0321] Each group contains 3 healthy male and female ICR mice aged 6-8 weeks.

[0322] The compound was prepared on the day of oral administration, using a solvent of 10% DMSO + 15% Solutol HS-15 and 0.5% sodium carboxymethyl cellulose aqueous solution (75%).

[0323] Mice were fasted overnight before administration and fed 4 hours after administration.

[0324] Mice were administered the drug by gavage. Blood samples were collected from the submandibular vein or other suitable veins at 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h post-administration. Approximately 0.02 mL of blood was collected per sample, anticoagulated with K2-EDTA, and placed on ice after collection. Plasma samples were separated within 1 hour after collection and placed on ice (centrifugation conditions: 6800 g, 6 minutes, 2-8 °C). Plasma samples were stored at -80 °C before analysis.

[0325] Samples were analyzed by LC-MS / MS, and the intraday accuracy of quality control samples was evaluated, with over 66.7% of the quality control samples required to have an accuracy between 80-120%. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on blood drug concentration data at different time points, providing parameters such as AUC0-t, AUC0-∞, MRT0-∞, Cmax, Tmax, and T1 / 2, along with their mean and standard deviation.

Claims

1. A compound of formula (I) and stereoisomers thereof, or a pharmaceutically acceptable salt thereof: wherein R1, R2, R3are each independently selected from H, halo, -Ci-C6alkyl, -C3-C6cycloalkyl, -Ci-C6haloalkyl, -OR a1 , -NR a2 R a3 ; or R2, R3and the attached C form a 5-10 membered heterocyclyl, 5-10 membered heteroaryl, which is optionally substituted with one or more halo, -Ci-C6alkyl, -C3-C6cycloalkyl, -Ci-C6haloalkyl, -OR a1 , -NR a2 R a3 substituted; R a1 , R a2 , R a3 each independently is selected from the group consisting of H, -Ci-C6alkyl, -Ci-C6haloalkyl, -C3-C6cycloalkyl; R4is selected from H, -Ci-C6alkyl; R5, R6, R7are each independently selected from H, halo, -Ci-C6alkyl, -Ci-C6alkoxy, -Ci-C6haloalkyl; A1, A2are each independently selected from -NR8, -CR9R 10 ; R8is selected from H, -Ci-C6alkyl, -Ci-C6alkoxy, -Ci-C6haloalkyl, -C3-C6cycloalkyl, -(CH2) r OC1-C6alkyl, -Ci-C6hydroxyalkyl, -(CH2) r -NR a2 R a3 , -C(O)Ci-C6alkyl; R9, R 10 each independently selected from H, -Ci-C6alkyl; m is 0, 1, 2, 3; n is 0, 1, 2, 3; r is 0, 1, 2, 3, 4.

2. The compound according to claim 1, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, The 5-10 membered heterocyclyl is 5-6 membered heterocyclyl, the 5-10 membered heteroaryl is 5-6 membered heteroaryl.

3. The compound according to claim 1, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, The compound is selected from:

4. The compound according to any one of claims 1 to 3, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, The compound has the structure shown below:

5. The compound according to any one of claims 1 to 4, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, R1is selected from H, halo, -Ci-C6alkyl, -C3-C6cycloalkyl.

6. The compound according to claim 5, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, R1is selected from H, methyl, chloro, cyclopropyl.

7. The compound according to any one of claims 1-6, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein, R2, R3are each independently selected from H, amino, ethyl, methyl, trifluoromethyl, cyclopropyl, methoxy, trifluoromethoxy, chloro, trifluoromethoxy.

8. The compound according to any one of claims 1-3, 5-7, and stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein, R4is selected from -Ci-C6alkyl, preferably from methyl.

9. The compound according to any one of claims 1-3, 5-7, and stereoisomers thereof, or a pharmaceutically acceptable salt thereof, wherein, m is 1.

10. The compound according to any one of claims 1-9, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein, R5, R6are each independently selected from H, -Ci-C6alkyl.

11. The compound according to claim 10, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, R5, R6are each independently selected from H.

12. The compound according to any one of claims 1-11, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein, R7is selected from H, -Ci-C6alkyl.

13. The compound according to claim 12, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, characterized in that, R7is selected from H, methyl.

14. The compound according to any one of claims 1-13, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein, R8is selected from methyl, ethyl, cyclopropyl, isopropyl, -CH2CH2F, -CH2CHF2, -(CH2)2OCH3, -(CH2)2OH, -(CH2)2N(CH3)2, -C(O)CH3, -CH2CH2OCH3.

15. A compound selected from the group consisting of: ###0009### and stereoisomers thereof, or a pharmaceutically acceptable salt thereof.

16. A pharmaceutical composition comprising a compound according to any one of claims 1 to 15, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers.

17. Use of a compound according to any one of claims 1 to 15, and stereoisomers thereof, or pharmaceutically acceptable salts thereof, or a pharmaceutical composition of claim 16, for the manufacture of a medicament for the treatment of a disease, disorder, syndrome, or condition associated with PRMT5 enzyme.

18. The use according to claim 17, wherein the disease, disorder, syndrome, or condition associated with PRMT5 enzyme is cancer, a hematological disease, an inflammatory disease, an autoimmune disease, a metabolic disease, a genetic disease, a hormone-related disease, an immunodeficiency disease, a cell death-related disease, a destructive bone disease, thrombin-induced platelet aggregation, a liver disease, and a cardiovascular disease.

19. The use according to claim 18, wherein the disease, disorder, syndrome, or condition associated with PRMT5 enzyme is cancer.

20. The use according to claim 19, wherein the cancer is advanced solid tumor, metastatic pancreatic cancer, metastatic non-small cell lung cancer, a neural tumor, adenocarcinoma, endometrioid carcinoma, metastatic esophageal cancer, metastatic head and neck cancer, squamous cell carcinoma, cervical tumor, myelodysplastic syndrome, non-Hodgkin lymphoma, acute myeloid leukemia, adenoid tumor, hematological tumor, melanoma, pancreatic tumor, brain tumor, glioblastoma, glioma, myelofibrosis, breast tumor, chronic myelomonocytic leukemia, diffuse large B-cell lymphoma, bladder cancer, cholangiocarcinoma, mesothelioma, ovarian cancer, lung cancer, prostate cancer, colon cancer, gastric cancer, esophageal cancer, and hepatocellular carcinoma.

21. A method of treating a MTAP-deficient and / or MTA-accumulating disease in a subject in need thereof by administering to the subject a therapeutically effective amount of a compound of any one of claims 1 to 15 or a pharmaceutical composition of claim 16.

22. The method of claim 21, wherein the disease is a proliferative disease.

23. The method of claim 22, the proliferative disease is a MTAP-deficient and / or MTA-accumulating cancer.

24. The method of claim 23, wherein the cancer is a glioblastoma, a malignant peripheral nerve sheath tumor (MPNST), an esophageal cancer (e.g., an esophageal squamous cell carcinoma or an esophageal adenocarcinoma), a bladder cancer (e.g., a bladder urothelial carcinoma), a pancreatic cancer (e.g., a pancreatic adenocarcinoma), a mesothelioma, a melanoma, a non-small cell lung cancer (NSCLC; e.g., a lung squamous or lung adenocarcinoma), a astrocytoma, an undifferentiated pleomorphic sarcoma, a diffuse large B-cell lymphoma (DLBCL), a leukemia, a head and neck cancer, a gastric adenocarcinoma, a myxofibrosarcoma, a cholangiocarcinoma, a brain cancer, a stomach cancer, a kidney cancer, a breast cancer, an endometrial cancer, a urinary tract cancer, a liver cancer, a soft tissue cancer, a pleural cancer, and a large intestine cancer or a sarcoma.

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