Compound for treating or preventing LRRK2 mediated diseases

Abstract

The present invention relates to a novel compound as shown in formula (I) and a pharmaceutically acceptable salt thereof, said compound being capable of inhibiting leucine-rich repeat kinase 2 (LRRK2), and a pharmaceutically acceptable salt thereof, said compound being capable of inhibiting leucine-rich repeat kinase 2 (LRRK2); the invention also relates to pharmaceutical compositions thereof, processes for their preparation and therapeutic uses.

Description

Technical field:

[0001] The present invention belongs to the field of pharmaceutical technology and specifically relates to a novel compound that inhibits LRRK2. The present invention provides the novel compound or a pharmaceutically acceptable salt thereof, as well as a pharmaceutically acceptable carrier, and uses thereof for treating or preventing LRRK2-mediated conditions, including neurodegenerative diseases such as Parkinson's disease, and immune-inflammatory diseases such as inflammatory bowel disease and glaucoma. Background of the invention:

[0002] Leucine-rich repeat kinase 2 (LRRK2) is a protein kinase encoded by the LRRK2 gene (PARK8) and a member of the ROCO protein family. It consists of 2527 amino acids (286 kDa) and contains multiple domains, including ARM, ANK, LRR, Roc, COR, Kinase, and WD40. The Roc and COR domains together constitute the GTPase domain. LRRK2 is a large, multifunctional protein with GTPase and serine-threonine kinase activities. Studies have shown that LRRK2 is expressed in multiple organs (including the brain, kidney, lung, liver, heart, and spleen), primarily in the cytoplasm and various membrane structures (including mitochondria, lysosomes, endosomes, lipid rafts, and vesicles), and is associated with multiple cellular functions (including autophagy, cytoskeletal dynamics, intracellular membrane trafficking, synaptic vesicle recycling, and inflammatory responses). Abnormal LRRK2 pathway is closely related to many diseases, including Parkinson's disease, glaucoma and inflammatory bowel disease.

[0003] Parkinson's disease (PD) is the second most common neurodegenerative disease after Alzheimer's disease. It is highly prevalent in middle-aged and elderly people (1-2%) and affects over 6 million people worldwide. The primary pathological features of PD are degeneration and loss of dopaminergic neurons in the substantia nigra and the formation of Lewy bodies. Clinical manifestations include motor symptoms such as resting tremor, bradykinesia, muscle rigidity, and postural balance disorders. These symptoms are often accompanied by sleep disorders, autonomic dysfunction, psychiatric symptoms such as depression, and non-motor symptoms such as cognitive impairment, which severely impact patients' quality of life. Current treatments focus on relieving symptoms, particularly movement disorders caused by dopamine depletion. Common therapeutic agents include compound dopa preparations, dopamine receptor agonists, and monoamine oxidase inhibitors. These drugs, through exogenous supplementation, potency enhancement, and metabolic inhibition, replace and replenish the function of depleted dopamine, and can alleviate motor symptoms to a certain extent in the early stages of the disease. However, these treatments only address the symptoms, failing to halt disease progression and largely ineffective for non-motor symptoms. They also suffer from significant side effects and diminishing efficacy. Therefore, there is an urgent clinical need for disease-modifying therapies that can halt or slow disease progression. Genome-wide association studies (GWAS) have shown that LRRK2 gene mutations, including major pathogenic mutations such as the G2019S mutation, are the most common cause of familial Parkinson's disease (PD). These mutations lead to overactivation of LRRK2. For example, the G2019S mutation increases LRRK2 protein kinase activity by 2-3 fold. Furthermore, multiple studies have shown that LRRK2 overactivation is also present in patients with idiopathic Parkinson's disease (PD). Therefore, LRRK2 inhibitors are widely considered to be promising disease-modifying therapies that can halt or slow disease progression by inhibiting overactivated LRRK2 kinase function. Currently, at least two LRRK2 inhibitors are in clinical trials for the treatment of Parkinson's disease.

[0004] Glaucoma is a group of diseases characterized by optic disc atrophy and depression, visual field loss, and decreased vision. Pathologically elevated intraocular pressure and insufficient blood supply to the optic nerve are primary risk factors for its development. The optic nerve's tolerance to pressure damage is also associated with the development and progression of glaucoma. Glaucoma is one of the three leading causes of blindness in humans, with an incidence of 1% in the general population and 2% after age 45. In China alone, there are over 20 million glaucoma patients. LRRK2 is widely distributed in TM cells. LRRK2 inhibitors can regulate LRRK2 in TM cells and may reduce intraocular pressure by inhibiting the contractile tension of the actin cytoskeleton, leading to TM relaxation. Currently, one LRRK2 inhibitor is in clinical trials for the treatment of glaucoma.

[0005] Inflammatory bowel disease (IBD) is a kind of idiopathic intestinal inflammatory disease involving ileum, rectum and colon. The main clinical manifestations of the disease are diarrhea, abdominal pain, hematochezia and weight loss, etc., including Crohn's disease (CD) and ulcerative colitis (UC). Ulcerative colitis is a continuous inflammation of the mucosal layer and submucosal layer of the colon, and the disease usually involves the rectum and gradually spreads to the entire colon. Crohn's disease can involve the entire digestive tract and is a non-continuous full-thickness inflammation, and the most common sites of involvement are the terminal ileum, colon and perianal region. The cause of inflammatory bowel disease is not fully understood, and it is known that the inflammatory response caused by abnormal response of the intestinal mucosal immune system plays an important role in the pathogenesis of inflammatory bowel disease. It is currently believed that this is caused by the interaction of multiple factors, mainly including environmental, genetic, infectious and immune factors. Genome-wide association studies have found that the LRRK2 gene is one of the main genetic loci affecting the genetic susceptibility of CD. Studies have shown that the expression of LRRK2 is up-regulated in the dendritic cells of Crohn's disease patients and the lymphoblastoid cell lines of patients carrying high-risk alleles. Analysis of mouse models shows that overexpression of LRRK2 can exacerbate colitis, which is related to Dectin-1-induced pro-inflammatory cytokine response in intestinal dendritic cells and autophagy defects. LRRK2 inhibitors can alleviate colitis, providing a new idea for the treatment of inflammatory bowel disease. Currently, LRRK2 inhibitors are in the preclinical discovery stage for the treatment of inflammatory bowel disease.

[0006] In addition, LRRK2 inhibitors are also a potential treatment for a variety of diseases, including tuberculosis, leprosy, Alzheimer's disease, dementia, Pick's disease, progressive supranuclear palsy, amyotrophic lateral sclerosis, neuroinflammation, multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, autoimmune hemolytic anemia, pure red cell anemia, idiopathic thrombocytopenic purpura, Evans syndrome, vasculitis, bullous skin disorder, type I diabetes, Sjogren's syndrome, Devic's disease, inflammatory myopathy, ankylosing spondylitis, glioblastoma, lymphoma, acute myeloid leukemia, renal cancer, breast cancer, lung cancer, prostate cancer, thyroid cancer, etc.

[0007] LRRK2 inhibitors have urgent clinical needs and broad application prospects. At present, some related literatures and patent applications have disclosed small molecule LRRK2 inhibitors, including WO2012062783A1, WO2014001973A1, WO2015092592A1, WO2016036586A1, WO2017046675A1, WO2017218843A1 and WO2020247298A3, etc., but there is still an urgent need to develop LRRK2 inhibitors with novel structure and high activity. The present application provides a pyridopyrrole derivative with a novel structure, and it is found that the compound with such structure has good LRRK2 inhibitory activity. SUMMARY

[0008] The present application provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0009]

[0010] wherein:

[0011] X is O, S, NH or CH2;

[0012] Y is a single bond, C(=O) or CHR 4 ;

[0013] R 1 is aryl or heteroaryl optionally substituted with 1 or 2 R 1a , said heteroaryl containing at least 1 N atom;

[0014] R 1a is hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylamino, hydroxyl, amino, cyano or halogen;

[0015] R 2 is hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 alkylamino, hydroxyl, amino, cyano or halogen;

[0016] R 3 is hydrogen, C 1-6 alkyl, C 1-6 alkoxy, hydroxyl or oxo;

[0017] R 4 is hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom;

[0018] n is 0, 1, or 2;

[0019] m is 0 or 1.

[0020] In some embodiments, X is O or CH2;

[0021] R 1 is phenyl, pyrrolyl or pyridyl;

[0022] R 1a It is hydrogen, C 1-6 Alkyl, cyano or halogen;

[0023] R 2 It is hydrogen, C 1-6 Alkyl or halogen;

[0024] R 3 is hydrogen or oxo;

[0025] R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl or optionally C 1-6 Alkyl-substituted pyrazolyl.

[0026] In some embodiments, R 1 is phenyl or pyrrolyl;

[0027] R 1a is hydrogen, methyl or cyano;

[0028] R 2 is hydrogen or a halogen;

[0029] R 4 is hydrogen, methyl, trifluoromethyl or pyrazolyl optionally substituted with methyl.

[0030] In some embodiments, the compound has the specified stereochemical formula (I'):

[0031]

[0032] In some embodiments, the compound has the chemical structure of formula (II),

[0033]

[0034] in:

[0035] Y is a single bond, C(=O) or CHR 4 ;

[0036] R 1 is optionally replaced by 1 or 2 R 1a a substituted aryl or heteroaryl group, wherein the heteroaryl group contains at least one nitrogen atom;

[0037] R 1a It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen;

[0038] R 2 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen;

[0039] R 3 It is hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy or oxo;

[0040] R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom;

[0041] n is 0, 1, or 2.

[0042] In some embodiments, the compound has the chemical structure of formula (III),

[0043]

[0044] in:

[0045] R 1 is optionally replaced by 1 or 2 R 1a a substituted aryl or heteroaryl group, wherein the heteroaryl group contains at least one nitrogen atom;

[0046] R 1a It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen;

[0047] R 2 It is hydrogen, C 1-6 Alkyl, C 1-6Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen;

[0048] R 3 It is hydrogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy or oxo;

[0049] R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom;

[0050] n is 0, 1, or 2.

[0051] In some embodiments, the compound has a chemical structure of Formula (IIa) or Formula (IIb),

[0052]

[0053] in:

[0054] R 1 is optionally replaced by 1 or 2 R 1a substituted phenyl or pyrrolyl;

[0055] R 1a It is hydrogen, C 1-6 Alkyl or cyano;

[0056] R 4 It is hydrogen, C 1-6 Alkyl or C 1-6 Halogenated alkyl.

[0057] In some embodiments, R 1 yes

[0058] R 4 is hydrogen, methyl or trifluoromethyl; preferably, R 4 It's hydrogen.

[0059] The present invention includes all pharmaceutically acceptable salt forms of the compounds. Such salts can be prepared using commercially available reagents by conventional organic chemical methods, and specifically include hydrochlorides, hydrobromides, hydroiodides, phosphates, sulfates, nitrates, formates, acetates, succinates, benzenesulfonates, citrates, glucuronates, lactates, methanesulfonates, toluenesulfonates, pamoates, and tartrates.

[0060] The present invention is intended to include all isotopes of atoms in all compounds. Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known in the art or by methods analogous to those described herein, and these compounds have the potential to improve pharmacological or pharmacokinetic properties.

[0061] As for stereoisomers, the compounds of the present invention may have chiral centers and may exist in the form of racemates, racemic mixtures, and individual enantiomers or diastereomers. All such isomeric forms, including mixtures thereof, are included in the present invention.

[0062] In addition, some of the crystalline forms of the compound of the present invention can exist in the form of polymorphs, which are also included in the present invention. In addition, some of the compounds can also form solvates with water or other organic solvents, and such solvates are also similarly included within the scope of the present invention.

[0063] On the other hand, the specific compounds provided by the present invention include but are not limited to the following compounds:

[0064]

[0065]

[0066] In another aspect, the present invention provides a process for preparing a compound of formula (I), wherein:

[0067] Compound 1-12 can be synthesized by the following scheme:

[0068]

[0069] Compound 13 can be synthesized by the following scheme:

[0070]

[0071] Compound 14 can be synthesized by the following scheme:

[0072]

[0073] In another aspect, the present invention provides pharmaceutical formulations suitable for use in human patients, comprising any of the compounds shown above (e.g., compounds of the present invention, such as compounds of formula (I)) and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical formulations can be used to treat or prevent the conditions or diseases described herein.

[0074] In another aspect, the present invention discloses the use of the compound in the preparation of a medicament for treating LRRK2 receptor-related disorders, including but not limited to neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease, spinocerebellar atrophy, Friedreich's ataxia, Pick's disease, dementia with Lewy bodies, dystonia, amyotrophic lateral sclerosis, neuroinflammation, progressive supranuclear palsy, and frontotemporal dementia; immune-inflammatory diseases such as inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, ankylosing spondylitis, autoimmune hemolytic anemia, pure red cell aplasia, idiopathic thrombocytopenic purpura, Evan's syndrome, vasculitis, bullous skin disorders, type I diabetes, Sjögren's syndrome, Devic's disease, and inflammatory myopathies; as well as malignant tumors and glaucoma.

[0075] Terminology

[0076] Alkyl or alkane is a fully saturated straight or branched chain non-aromatic hydrocarbon. Typically, unless otherwise defined, a straight or branched chain alkyl group has 1 to about 20 carbon atoms, preferably 1 to about 10 carbon atoms. Examples of straight and branched chain alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, amyl and octyl. C1-C6 straight or branched chain alkyl groups are also referred to as "lower alkyl groups."

[0077] Furthermore, the term "alkyl" (or "lower alkyl") as used throughout the specification, examples, and claims is intended to include "unsubstituted alkyl" and "substituted alkyl," the latter referring to alkyl moieties having substituents replacing hydrogen on one or more carbons of the hydrocarbon backbone. If not otherwise specified, such substituents may include, for example, halogen, hydroxy, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as thioester, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or aromatic or heteroaromatic moieties. Those skilled in the art will understand that the moieties substituted on the hydrocarbon chain may themselves be substituted, if appropriate. For example, substituents of a substituted alkyl group may include amino, azido, imino, amide, phosphoryl (including phosphonate and phosphite), sulfonyl (including sulfate, sulfonamide, sulfamoyl and sulfonate), and silyl, as well as substituted and unsubstituted forms of ether, alkylthio, carbonyl (including ketone, aldehyde, carboxylate and ester), -CF3, -CN, and the like.

[0078] The cycloalkyl group preferably has 3 to 7 ring carbon atoms and can be substituted or unsubstituted. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, and methylcyclopentyl. The cycloalkyl group can be further substituted with alkyl, alkenyl, alkoxy, alkylthio, aminoalkyl, carbonyl-substituted alkyl, -CF , -CN, and the like.

[0079] Aryl represents a monocyclic or bicyclic fused-ring aromatic group having 5 to 10 carbon atoms, such as phenyl, 1-naphthyl or 2-naphthyl; or a partially saturated bicyclic fused-ring containing a phenyl group, such as indanyl, dihydro- or tetrahydronaphthyl.

[0080] Heteroaryl refers to an aromatic group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur and is monocyclic or bicyclic. Monocyclic heteroaryl includes 5-8 membered heteroaryl groups containing 1, 2, 3, or 4 heteroatoms selected from nitrogen, sulfur, and oxygen. Bicyclic heteroaryl includes 9 or 10 membered fused ring heteroaryl groups. Heteroaryl groups include, for example, pyrrolyl, thienyl, furyl, pyrazolyl, imidazolyl, triazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, and benzo-fused derivatives of such monocyclic heteroaryl groups such as indolyl, benzimidazolyl, or benzofuranyl, quinolyl, isoquinolyl, quinazolinyl, or purinyl.

[0081] In optionally substituted heteroaryl, the substituents are preferably lower alkyl, lower alkoxy, lower alkoxy-lower alkoxy, amino, optionally substituted by one or two substituents selected from lower alkyl, lower alkenyl and alkylcarbonyl, halo-lower alkyl, lower alkoxy-lower alkyl, halogen or nitro.

[0082] Alkenyl contains one or more, for example two or three, double bonds and is preferably lower alkenyl, for example 1- or 2-butenyl, 1-propenyl, allyl or vinyl.

[0083] Alkynyl is preferably a lower alkynyl group such as propargyl or ethynyl.

[0084] In optionally substituted alkenyl or alkynyl groups, the substituents are preferably lower alkyl, lower alkoxy, halo or di(lower alkyl)amino and are attached to a saturated carbon atom of the alkenyl or alkynyl group or to an unsaturated carbon atom of the alkenyl group.

[0085] Heterocyclyl preferably means a saturated, partially saturated or unsaturated monocyclic or bicyclic ring containing 4 to 10 atoms, including 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, which may be carbon- or nitrogen-linked, unless otherwise specified, wherein the ring nitrogen atoms may be optionally substituted by groups selected from lower alkyl, amino-lower alkyl, aryl, aryl-lower alkyl and acyl, and the ring carbon atoms may be substituted by lower alkyl, amino-lower alkyl, aryl, aryl-lower alkyl, heteroaryl, lower alkoxy, hydroxy or oxo. Heterocyclyl is exemplified by pyrrolidinyl, oxazolidinyl, thiazolidinyl, piperidinyl, morpholinyl, piperazinyl, dioxolanyl or tetrahydropyranyl.

[0086] Acyl represents, for example, alkylcarbonyl, cyclohexylcarbonyl, arylcarbonyl, aryl-lower alkylcarbonyl or heteroarylcarbonyl. Lower acyl is preferably lower alkylcarbonyl, in particular propionyl or acetyl.

[0087] The hydroxyalkyl group refers to an alkyl group substituted with at least one hydroxy group, preferably a hydroxy-lower alkyl group, such as hydroxymethyl, 2-hydroxyethyl, 2-hydroxy-n-propyl, and hydroxyisopropyl.

[0088] The cyanoalkyl group refers to an alkyl group substituted by at least one cyano group, preferably a cyano-lower alkyl group, such as cyanomethyl or cyanoethyl.

[0089] The haloalkyl group refers to an alkyl group substituted by at least one halogen, preferably a halo-lower alkyl group, such as monofluoromethyl, difluoromethyl, trifluoromethyl, 3,3,3-trifluoroethyl or pentafluoroethyl.

[0090] Halogen is fluorine, chlorine, bromine or iodine.

[0091] Lower alkoxy is especially methoxy, ethoxy, isopropoxy or tert-butoxy.

[0092] Arylalkyl includes aryl and alkyl as defined above and is, for example, benzyl, 1-phenylethyl or 2-phenylethyl.

[0093] Heteroarylalkyl includes heteroaryl and alkyl as defined above and is, for example, 2-, 3- or 4-pyridylmethyl, 1- or 2-pyrrolylmethyl, 1-pyrazolylmethyl, 1-imidazolylmethyl, 2-(1-imidazolyl)ethyl or 3-(1-imidazolyl)propyl.

[0094] Two adjacent substituents which, together with the atoms of the aryl or heteroaryl radicals, can form a 5- or 6-membered carbocyclic or heterocyclic ring are, for example, propylene, 1- or 2-oxopropylene, 1- or 2-oxapropylene, 1-oxapropylidene, methylenedioxy, difluoro-methylenedioxy, 1- or 2-azapropylene, 1- or 2-azapropylidene, 1,2- or 1,3-diaza-propylidene, 1,3-diaza-2-oxopropylene, butylene, 1- or 2-oxabutylene, ethylenedioxy, 1- or 2-azetidinyl or 1- or 2-azabutadienyl or such radicals which carry further substituents as defined above. Specific implementation method:

[0095] Synthesis Example

[0096] In order to make the purpose and technical solutions of the present invention clearer, the present invention is further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, the specific experimental methods not mentioned in the following examples are all carried out according to conventional experimental methods.

[0097] The abbreviations used in this document have the following meanings:

[0098] Abbreviation meaning

[0099] LCMS liquid chromatography-mass spectrometry

[0100] Prep-HPLC

[0101] DMF N,N-dimethylformamide

[0102] PE petroleum ether

[0103] EA Ethyl acetate

[0104] THF Tetrahydrofuran

[0105] ACN acetonitrile

[0106] DCM dichloromethane

[0107] DMSO dimethyl sulfoxide

[0108] IPA Isopropyl alcohol

[0109] TFA trifluoroacetic acid

[0110] SEMCl 2-(Trimethylsilyl)ethoxymethyl chloride

[0111] NBS N-bromosuccinimide

[0112] NCS N-chlorosuccinimide

[0113] TMSCF3 (trifluoromethyl)trimethylsilane

[0114] TBAF Tetrabutylammonium fluoride

[0115] TsCl p-Toluenesulfonyl chloride

[0116] DI petroleum ether AN, N-diisopropylethylamine

[0117] DCC N,N'-dicyclohexylcarbodiimide

[0118] DMAP 4-dimethylaminopyridine

[0119] PMBCl 4-Methoxybenzyl chloride

[0120] BINAP 1,1'-binaphthyl-2,2'-bis(diphenylphosphine)

[0121] Pd2(dba)3 tris(dibenzylidene-base acetone)dipalladium

[0122] Pd(dppf)Cl2 1,1-bis(diphenylphosphine)diphenylferric palladium dichloride

[0123] Pd(dtbpf)Cl2 [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride

[0124] SFC Supercritical Fluid Chromatography

[0125] HPLC high-performance liquid chromatography

[0126] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance spectroscopy ( 1 H-NMR) or mass spectrometry (MS).

[0127] 1 H-NMR measurements were performed on a Bruker 400 MHz nuclear magnetic resonance instrument. The solvents used were deuterated methanol (CD3OD), deuterated chloroform (CDCl3), or hexadeuterated dimethyl sulfoxide (DMSO-d6), and the internal standard was tetramethylsilane (TMS). Chemical shifts (δ) are given in parts per million (ppm).

[0128] The mass spectrometry (MS) was performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0129] Thin layer chromatography (TLC) was performed using aluminum plates (20×20 cm) manufactured by Merck, and preparative thin layer chromatography was performed using GF254 (0.4-0.5 mm) silica gel plates.

[0130] The monitoring of the reaction was carried out by thin layer chromatography (TLC) or liquid chromatography-mass spectrometry (LC-MS), and the developing agent systems used included dichloromethane and methanol system, n-hexane and ethyl acetate system, and petroleum ether and ethyl acetate system. The developing agent system was adjusted (by adjusting the volume ratio of the solvents or adding triethylamine, etc.) according to the polarity of the compound to be separated.

[0131] Unless otherwise specified, the reaction temperature is room temperature (20-30°C).

[0132] A single bond representing a single chiral isomer.

[0133] The reagents used in the examples were purchased from Acros Organics, Aldrich Chemical Company, Shanghai Tebo Chemical Technology Co., Ltd., etc.

[0134] Synthesis of intermediate 1 (INT1):

[0135]

[0136] First step: To a solution of compound INT1-1 (9.3 g, 51.5 mmol) in DMF (200 mL), add CS2CO3 (42 g, 128.8 mmol), slowly drop 2-(trimethylsilyl)ethoxymethyl chloride (10.3 g, 61.8 mmol), and stir the reaction at room temperature. After the reaction is complete, slowly pour the reaction system into a NH4Cl solution (500 mL), extract with ethyl acetate (500 mL x 3), wash with saturated brine (300 mL x 3), dry over anhydrous sodium sulfate, filter, concentrate, and separate the crude product by column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain white solid compound INT1-2 (17.6 g, 73.0%). LCMS (ESI, m / z): 310.9 [M+H] + . 1 HNMR (400 MHz, CDCl3) δ 10.55 (s, 1H), 8.83 (s, 1H), 7.45 (s, 1H), 6.77 (s, 1H), 5.68 (s, 2H), 3.52 (t, J = 7.9 Hz, 2H), 0.90 (t, J = 7.8 Hz, 2H), -0.07 (s, 9H).

[0137] Step 2: To a solution of compound INT1-2 (12 g, 38.6 mmol) in acetonitrile (120 mL) was added N-bromosuccinimide (7.6 g, 42.5 mmol) in portions and stirred at room temperature. The reaction system was poured into a saturated NaHCO3 solution (200 mL), extracted with ethyl acetate (200 mL x 3), washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated by column chromatography to obtain compound INT-1 (11.3 g, yield 75.1%) as a white solid. LCMS (ESI, m / z): 390.7 [M+H] + . 1 H NMR (400MHz, CDCl3) δ10.59(s,1H),8.81(s,1H),7.50(s,1H),5.66(s,2H),3.64-3.47(m,2H),0.95-0.85(m,2H),-0.01--0.14(m,9H).

[0138] Example 1: Synthesis of (S)-3-(3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxirane-1-yl)benzonitrile (Compound 1)

[0139]

[0140] Step 1: To a solution of compound INT1 (2.240 g, 5.758 mmol) in isopropanol (30 mL) were added (S)-2-(((tert-butyldimethylsilyl)oxy)methyl)pyrrolidine (1.868 g, 8.648 mmol) and diisopropylethylamine (1.486 g, 11.519 mmol), and the mixture was stirred at 70°C for 2 hours. After completion of the reaction, the reaction solution was concentrated and separated by silica gel column chromatography (petroleum ether:ethyl acetate = 7:1) to obtain compound 1-2 (2.0 g, yield 61.1%) as a brown oil. LCMS (ESI, m / z): 568.3 [M+H] + . 1H NMR (400MHz, CDCl3) δ10.42(s,1H),8.73(s,1H),7.40(s,1H),5.72-5.63(m,2H),4.36-4.27(m,1H),3.81-3.75(m,1H),3.62-3.57(m,2H) ,3.53-3.46(m,3H),2.44-2.33(m,1H),2.20-2.06(m,2H),1.93-1.83 (m,1H),0.98-0.95(m,2H),0.69(s,9H),-0.00(s,9H),-0.13(s,6H).

[0141] Step 2: Sodium borohydride (0.263 mg, 6.972 mmol) was added to a solution of compound 1-2 (1.98 g, 3.486 mmol) in ethanol (30 mL) and stirred at room temperature for 4 hours. After completion of the reaction, saturated aqueous ammonium chloride (30 mL) was slowly added for dilution. The mixture was extracted with ethyl acetate (40 mL x 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and separated by silica gel column chromatography (petroleum ether:ethyl acetate = 7:1) to obtain compound 1-3 (1.72 g, yield 86.5%) as a yellow oil. LCMS (ESI, m / z): 572.3 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.32 (s, 1H), 7.42-7.37 (m, 1H), 5.66 (q, J = 10.8Hz, 2H) ,5.03-4.91(m,1H),4.60(d,J=9.7Hz,1H),4.38-4.22(m,2H),3.64-3.56(m,5 H),3.49-3.37(m,1H),2.45-2.32(m,1H),2.17-2.09(m,2H),1.80-1.72(m,1H ),0.99-0.93(m,2H),0.72(s,9H),-0.00(s,9H),-0.10(s,3H),-0.20(s,3H).

[0142] Step 3: To a solution of compound 1-3 (1.65 g, 2.889 mmol) in tetrahydrofuran (25 mL) and water (5 mL) were added 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (0.86 g, 3.756 mmol), Pd(dtbpf)Cl2 (189 mg, 0.289 mmol), and potassium phosphate (1.229 g, 5.779 mmol). The mixture was stirred at 60°C for 2 hours. After completion, the reaction was diluted with water (40 mL) and extracted with ethyl acetate (40 mL x 3). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and separated by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain compound 1-4 (1.63 g, 95.1% yield) as a yellow oil. LCMS (ESI, m / z): 593.5 [M+H] + . 1 H NMR (400MHz, CDCl3) δ8.25 (s, 1H), 7.82-7.78 (m, 1H), 7.76-7.70 (m, 1H), 7.63 (d, J = 7.8Hz, 1H), 7.52 (t, J = 7. 7Hz,1H),7.29(s,1H),5.71(q,J=10.7Hz,2H),4.99(dd,J=12.2,3.6Hz,1H),4.66(dd,J=12.1,6.5Hz,1H),3. 79-3.69(m,1H),3.68-3.59(m,2H),3.52(s,1H),3.48-3.43(m,1H),3.43-3.32(m,2H),3.31-3.21(m,1H),1. 96-1.84(m,2H),1.39-1.30(m,2H),1.03-0.94(m,2H),0.69(s,9H),0.00(s,9H),-0.13(s,3H),-0.23(s,3H).

[0143] Step 4: To a solution of compound 1-4 (400 mg, 0.675 mmol) in dichloromethane (8 mL) was added thionyl chloride (161 mg, 1.349 mmol) and stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated and purified via a reverse phase column (acetonitrile:H2O(NH3.H2O) = 3:2) to afford compound 1-5 as a yellow solid (100 mg, 32% yield). LCMS (ESI, m / z): 461.3 [M+H] + . 1H NMR(400MHz, CDCl3)δ8.14(s,1H),7.82-7.78(m,1H),7.77-7.73(m,1H),7.65-7.60(m,1H), 7.53(t,J=7.7Hz,1H),7.33(s,1H),5.73(q,J=10.6Hz,2H),5.07(d,J=13.9Hz,1H),4.51(d,J =13.8Hz,1H),4.00-3.86(m,2H),3.70-3.59(m,2H),3.33(t,J=11.4Hz,1H),3.06-2.98(m,1H ),2.94-2.85(m,1H),1.40-1.25(m,3H),1.02-0.95(m,2H),0.93-0.88(m,1H),-0.00(s,9H).

[0144] Step 5: Add trifluoroacetic acid (1 mL) to a solution of compound 1-5 (90 mg, 0.195 mmol) in dichloromethane (1 mL), stir at room temperature for 4 hours, concentrate to remove the solvent, and add ammonia methanol solution (4 mL, 7 M) to the residue, stirring at room temperature for 5 hours. After the reaction is complete, the reaction solution is concentrated and reversed phase to obtain compound 1. LCMS (ESI, m / z): 330.9 [M+H] + . 1 H NMR (400MHz, DMSO) δ11.89(s,1H),8.02(s,1H),7.86(s,1H),7.80(d,J=7.8Hz ,1H),7.71(d,J=7.7Hz,1H),7.63-7.56(m,2H),4.96(d,J=13.8Hz,1H),4.40( d,J=13.7Hz,1H),3.90-3.75(m,2H),3.24(t,J=11.0Hz,1H),2.93-2.85(m,1H ),2.78-2.70(m,1H),2.11-2.02(m,1H),1.67-1.53(m,2H),1.30-1.17(m,1H).

[0145] Example 2: Synthesis of (R)-3-(3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxirane-1-yl)benzonitrile (Compound 2)

[0146]

[0147] Compound 2 was prepared according to the procedure described in Example 1 except that (R)-2-(((tert-butyldimethylsilyl)oxy)methyl)pyrrolidine was used in the first step instead of the enantiomer (S)-2-(((tert-butyldimethylsilyl)oxy)methyl)pyrrolidine used in the first step of Example 1. LCMS (ESI, m / z): 331.1 [M+H] + . 1 H NMR (400 MHz, DMSO) δ

[0148] 11.89 (s, 1H), 8.01 (s, 1H), 7.86 (s, 1H), 7.80 (d, J = 7.8 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.64 - 7.55 (m, 2H), 4.96 (d, J = 13.8 Hz, 1H), 4.40 (d, J = 13.7 Hz, 1H), 3.92 - 3.76 (m, 2H), 3.28 - 3.21 (m, 1H), 2.93 - 2.85 (m, 1H), 2.79 - 2.70 (m, 1H), 2.16 - 2.00 (m, 1H), 1.69 - 1.50 (m, 2H), 1.31 - 1.16 (m, 1H).

[0149] Example 3: Synthesis of (S)-3-(10,10-difluoro-3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1- c]pyrrolo[3',2':5,6]pyrido[4,3-e][l,4]oxazocan-l-yl)benzonitrile (Compound 3)

[0150]

[0151] Compound 3-4 was prepared according to the procedure described in Example 1 except that (S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4,4-difluoropyrrolidine was used in the first step instead of (S)-2-(((tert-butyldimethylsilyl)oxy)methyl)pyrrolidine used in the first step of Example 1 as a light yellow liquid (300 mg, 32.6% overall yield for three steps). LCMS (ESI, m / z): 629.3 [M+H] + . 1H NMR (400MHz, DMSO-d6) δ8.19(s,1H),7.97(s,1H),7.83(d,J=7.5Hz,1H),7.76(d,J= 9.0Hz,2H),7.58(t,J=7.8Hz,1H),5.62(s,2H),5.16(t,J=5.1Hz,1H),4.70-4.55(m, 2H),3.94(s,2H),3.56-3.51(m,2H),3.51(s,1H),2.14-1.94(m,2H),1.23(d,J=4.3H z,2H),0.86-0.81(m,2H),0.63(s,9H),-0.08--0.10(m,9H),-0.23(t,J=6.0Hz,6H).

[0152] Step 4: To a solution of compound 3-4 (280 mg) in tetrahydrofuran (5 mL) was added 5 drops of concentrated sulfuric acid at 0°C. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was slowly added to 20 mL of water and the pH was adjusted to 8-9 with 2M sodium hydroxide. The mixture was extracted with ethyl acetate (20 mL x 2). The organic phase was washed with 10 mL of saturated sodium chloride and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to obtain a yellow solid compound 3-5 (230 mg, crude product). LCMS (ESI, m / z): 497.1 [M+H] + .

[0153] Step 5: Dissolve compound 3-5 (230 mg) in DCM (3 mL) and trifluoroacetic acid (3 mL) and stir at room temperature for 4 h. After completion of the reaction, the reaction solution was concentrated to dryness to obtain a reddish-brown liquid. The above residue was dissolved in MeOH·NH3 (7 M, 5 mL) and stirred at room temperature for 2 h. After completion of the reaction, the reaction solution was concentrated to dryness. The crude product was purified by preparative HPLC to obtain compound 3. LCMS (ESI, m / z): 367.2 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.11 (s, 1H), 7.98 (s, 1H), 7.86 (d, J = 7.8 Hz, 1H), 7.74 (d, J = 7.7 Hz, 1H), 7.63 (dd, J = 14.6, 6.8 Hz, 2H), 4.96 (d, J = 13.9 Hz, 1H), 4.48 (d, J = 13.9 Hz, 1H), 4.10 (t, J = 8.8 Hz, 1H), 3.85 (dd, J = 12.0, 2.7 Hz, 1H), 3.40 (dt, J = 22.9, 9.2 Hz, 2H), 2.97 (dd, J = 28.4, 13.1 Hz, 1H), 2.62 (ddd, J = 27.9, 14.5, 7.3 Hz, 1H), 2.21 (dd, J = 18.5, 14.6 Hz, 1H). 19 F NMR (377 MHz, DMSO-d6) δ -85.41 (s), -86.02 (s), -95.99 (s), -96.60 (s).

[0154] Example 4: Synthesis of (R)-3-(3,6,8a,9,11,12-hexahydro-8H-[1,4]oxazepino[3,4-c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxazepine-1-yl)benzonitrile (Compound 4)

[0155]

[0156] Compound 4 was prepared according to the procedure described in Example 3 except that (S)-3-((tert-butyldimethylsilyl)oxy)methyl)morpholine was used in the first step instead of (S)-2-(((tert-butyldimethylsilyl)oxy)methyl)-4,4-difluoropyrrolidine used in the first step in Example 3. LCMS (ESI, m / z): 347.1 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 11.88 (s, 1H), 7.95 (s, 1H), 7.92 (s, 1H), 7.84 (d, J = 7.8 Hz, 1H), 7.77 (d, J = 7.7 Hz, 1H), 7.64 (t, J = 7.7 Hz, 1H), 7.52 (s, 1H), 5.04 (d, J = 12.4 Hz, 1H), 4.66 (d, J = 14.2 Hz, 1H), 4.37 (t, J = 10.3 Hz, 1H), 3.92 (dd, J = 18.0, 7.7 Hz, 2H), 3.65 (t, J = 13.7 Hz, 2H), 3.16 (s, 1H), 3.06 (t, J = 10.8 Hz, 1H), 2.44 (s, 1H), 2.26 (s, 1H).

[0157] Example 5: Synthesis of (S)-4-(3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6]pyrido[4,3- e][l,4]oxazinan-l-yl)-l-methyl-lH-pyrrole-2-carbonitrile (Compound 5)

[0158]

[0159] Compound 5 was prepared according to the procedures described in Example 1 except that 1-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-lH-pyrrole-2-carbonitrile was used in place of 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzonitrile in the third step of Example 1. LCMS (ESI, m / z): 334.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.56 (s, 1H), 7.94 (s, 1H), 7.28 (s, 1H), 7.22 (d, J = 1.4 Hz, 1H), 6.99 (d, J = 1.6 Hz, 1H), 4.91 (d, J = 13.7 Hz, 1H), 4.37 (d, J = 13.6 Hz, 1H), 3.79 (s, 3H), 3.78 - 3.72 (m, 2H), 3.22 (t, J = 11.8 Hz, 1H), 3.14 - 3.01 (m, 1H), 3.02 - 2.92 (m, 1H), 2.15 - 2.05 (m, 1H), 1.75 - 1.62 (m, 1H), 1.62 - 1.50 (m, 1H), 1.50 - 1.38 (m, 1H).

[0160] Example 6: Synthesis of 3-((6R,8aS)-6-methyl-3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1- c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxazepin-1-yl)benzonitrile and 3-((6S,8aS)-6-methyl- 3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxazepin-1- yl)benzonitrile (Compounds 6 and 7)

[0161]

[0162] First Step: To the solution of compound 1-2 (968 mg, 1.702 mmol) in tetrahydrofuran (20 ml), nitrogen was bubbled, then MeMgBr (8.5 ml, 1 mol / L) was added into the reaction system, which was stirred at room temperature for 4 hours. After the reaction was completed, it was diluted with water (30 mL), extracted with ethyl acetate (40 mL x 3), washed with saturated brine (40 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 6-2 (629 mg, yield 63.2%) as a light yellow oil. LCMS (ESI, m / z): 584.1 [M+H] + .

[0163] Second Step: To the solution of compound 6-2 (540 mg, 0.923 mmol) in dioxane / water (10 / 1, 5 mL), compound 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (275 mg, 1.2 mmol), Pd(dtbpf)Cl2 (59.6 mg, 0.092 mmol) and potassium phosphate (588.1 mg, 2.77 mmol) were added, which was stirred at 80°C for 4 hours under nitrogen protection. After the reaction was completed, it was diluted with water (70 mL), extracted with ethyl acetate (70 mL x 3), washed with saturated brine (70 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound 6-3 (417 mg, yield 74.43%) as a light yellow oil. LCMS (ESI, m / z): 607.3 [M+H] + .

[0164] Third Step: To the solution of compound 6-3 (100 mg, 0.17 mmol) in ethyl acetate (1 mL), 5 drops of H2SO4 were added, which was stirred at room temperature for 2 hours. After the reaction was completed, it was concentrated to obtain the crude product (100 mg, Crude).

[0165] The above crude product was added to a solution of NH3-MeOH (2 mL) and stirred at room temperature for 4 hours. After the reaction was completed, it was diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by preparative high performance liquid chromatography to obtain compound 6-4. Compound 6-4 was a mixture of diastereoisomers, which were separated by supercritical fluid chromatography to obtain two isomers, compounds 6 and 7, respectively.

[0166] LCMS (ESI, m / z): 345.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.83 (s, 1H), 7.97 (s, 1H), 7.84 (s, 1H), 7.79 (d, J = 7.8 Hz, 1H), 7.70 (d, J = 7.7 Hz, 1H), 7.62-7.54 (m, 2H), 5.25 (q, J = 6.6 Hz, 1H), 3.99 (d, J = 7.3 Hz, 1H), 3.70 (q, J = 7.4, 5.6 Hz, 2H), 2.85 (q, J = 8.3 Hz, 1H), 2.75 (td, J = 8.6, 4.4 Hz, 1H), 2.17-1.93 (m, 1H), 1.59 (t, J = 8.2 Hz, 2H), 1.39 (d, J = 6.7 Hz, 3H), 1.31-1.03 (m, 1H).

[0167] LCMS (ESI, m / z): 345.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.89 (s, 1H), 8.14 (s, 1H), 7.83 (s, 1H), 7.78 (d, J = 7.9 Hz, 1H), 7.72 (d, J = 7.8 Hz, 1H), 7.63-7.56 (m, 2H), 4.49 (q, J = 6.8 Hz, 1H), 3.76 (dd, J = 11.6, 3.3 Hz, 1H), 3.63 (t, J = 9.5 Hz, 1H), 3.22 (d, J = 11.0 Hz, 1H), 2.88 (td, J = 8.3, 4.2 Hz, 1H), 2.68 (q, J = 7.4 Hz, 1H), 2.09-1.98 (m, 1H), 1.67 (d, J = 6.6 Hz, 4H), 1.59 (d, J = 9.6 Hz, 1H), 1.28-1.17 (m, 1H).

[0168] Example 8: Synthesis of 3-((6R,8aS)-6-(trifluoromethyl)-3,6,8a,9,10,11- hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6]pyrido[4,3-e][1,4]oxazepin-1-yl) and 3- ((6S,8aS)-6-(trifluoromethyl)-3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1-c]pyrrolo[3',2':5,6] pyrido[4,3-e][1,4]oxazepin-1-yl)benzonitrile (Compounds 8 and 9)

[0169]

[0170] First step: To a solution of compound 1-2 (2 g, 3.516 mmol) in tetrahydrofuran (20 ml), (trifluoromethyl)trimethylsilane (2 g, 7.034 mmol) was added, which was stirred at room temperature for 30 minutes, and then the reaction system was placed in an ice bath and tetrabutylammonium fluoride (1 g, 3.87 mmol) was added. After the reaction was completed, water (40 mL) was added for dilution, and ethyl acetate (40 mL x 3) was extracted, and saturated brine (40 mL x 3) was washed, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain compound 8-2 (544 mg, yield 29.5%) as a light yellow oil. LCMS (ESI, m / z): 524.9 [M+H] + .

[0171] Second step: To a solution of compound 8-2 (650 mg, 1.247 mmol) in tetrahydrofuran (6 mL), CS2CO3 (1.2 g, 3.74 mmol), p-toluenesulfonyl chloride (285.3 mg, 1.496 mmol) were added, which was stirred at room temperature overnight. After the reaction was completed, saturated ammonium chloride solution (40 mL) was added for quenching, and ethyl acetate (40 mL x 3) was extracted, and saturated brine (40 mL x 3) was washed, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was separated by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain compound 8-3 (148 mg, yield 23.44%) as a light yellow oil. LCMS (ESI, m / z): 507.4 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 8.14 (d, J = 8.3 Hz, 1H), 7.82 (d, J = 23.7 Hz, 1H), 7.74 - 7.63 (m, 1H), 5.56 (dd, J = 15.9, 9.8 Hz, 2H), 4.22 (t, J = 6.6 Hz, 2H), 4.10 (s, 1H), 3.85 (s, 1H), 3.77 - 3.71 (m, 2H), 3.21 (dd, J = 15.8, 9.0 Hz, 1H), 1.68 - 1.56 (m, 2H), 1.37 (dd, J = 14.9, 7.3 Hz, 2H), 0.91 (t, J = 7.4 Hz, 2H), -0.10 (d, J = 4.3 Hz, 9H).

[0172] Third Step: To a solution of compound 8-3 (120 mg, 0.23 mmol) in dioxane / H2O (10 / 1, 5 mL) was added 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzonitrile (69 mg, 0.30 mmol), Pd(dtbpf)Cl2 (15.3 mg, 0.023 mmol) and potassium phosphate (147 mg, 0.69 mmol), and the reaction mixture was stirred at 80 °C for 3 h under nitrogen atmosphere. After completion of the reaction, the reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (15 mL x 3). The combined organic extracts were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated to give the crude compound 8-4 (186 mg, crude). LCMS (ESI, m / z): 529.1 [M+H] + .

[0173] Fourth Step: To a solution of compound 8-4 (174 mg, 0.329 mmol) in trifluoroacetic acid (1.5 mL) was added and the reaction mixture was stirred at room temperature for 2 h. After completion of the reaction, the reaction mixture was concentrated with acetonitrile twice to give the crude product (174 mg, crude). LCMS (ESI, m / z): 429.8 [M+H] + .

[0174] To the above crude product was added NH3-MeOH (2 mL) and the reaction mixture was stirred at room temperature for 4 h. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic extracts were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by preparative high performance liquid chromatography to give compound 8-5. Compound 8-5 was a mixture of diastereomers which were separated by supercritical fluid chromatography to give two isomers, compound 8 and compound 9.

[0175] LCMS (ESI, m / z): 399.1 [M+H]+ . 1 H NMR (400 MHz, CDC13) δ 10.66 (s, 1H), 8.34 (s, 1H), 7.70 (s, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.33 (s, 1H), 4.88 (q, J = 7.7 Hz, 1H), 4.01 (dd, J = 11.8, 3.2 Hz, 1H), 3.82 (dd, J = 12.1, 6.6 Hz, 1H), 3.56 - 3.33 (m, 1H), 3.05 (td, J = 8.3, 3.3 Hz, 1H), 2.86 (dd, J = 16.2, 7.9 Hz, 1H), 2.15 - 2.08 (m, 1H), 1.71 - 1.60 (m, 2H), 1.48 - 1.38 (m, 1H).

[0176] LCMS (ESI, m / z): 399.1 [M+H] + . 1 H NMR (400 MHz, CDC13) δ 10.66 (s, 1H), 8.34 (s, 1H), 7.70 (s, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.33 (s, 1H), 4.88 (q, J = 7.7 Hz, 1H), 4.01 (dd, J = 11.8, 3.2 Hz, 1H), 3.82 (dd, J = 12.1, 6.6 Hz, 1H), 3.56 - 3.33 (m, 1H), 3.05 (td, J = 8.3, 3.3 Hz, 1H), 2.86 (dd, J = 16.2, 7.9 Hz, 1H), 2.15 - 2.08 (m, 1H), 1.71 - 1.60 (m, 2H), 1.48 - 1.38 (m, 1H).

[0177] Example 10: Synthesis of (S)-3-(6-(l-methyl-lH-pyrazol-4-yl)-3,6,8a,9,10,11- hexahydro-8H-pyrrolo[2,l-c]pyrrolo[3',2':5,6]pyrido[4,3-e][l,4]oxazepin-l-yl)benzonitrile (Compound 10)

[0178]

[0179] First step: To a solution of 4-iodo-l-methyl-lH-pyrazole (4.8 g, 23.0 mmol) in tetrahydrofuran (15 mL) was added at 0 °C iPrMgCl (23 mL, 1 M, 23.0 mmol) was added dropwise to THF (23 mL) at 0 °C. The reaction was stirred at room temperature for 2 h. Then a solution of compound 1-2 (1.308 g, 2.299 mmol) in THF (5 mL) was added, and the reaction was stirred at room temperature for another 2 h. After completion of the reaction, the reaction mixture was poured into 20 mL of saturated ammonium chloride solution and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2: 1) to give compound 10-2 (854 mg, 57.1%) as a yellow oil. LCMS (ESI, m / z): 652.1 [M+H] + .

[0180] Second step: To a mixture of compound 10-2 (854 mg, 1.312 mmol) in THF / H2O (10 / 1, 10 mL) was added 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzonitrile (451 mg, 1.968 mmol), Pd(dtbpf)Cl2 (85.5 mg, 0.131 mmol) and K3PO4 (836 mg, 3.937 mmol), and the reaction was stirred at 60 °C for 4 h under nitrogen protection. After completion of the reaction, the reaction mixture was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 1) to give compound 10-3 (476 mg, 53.9%) as a yellow solid. LCMS (ESI, m / z): 674.3 [M+H] + .

[0181] Third step: To a solution of compound 10-3 (200 mg, 0.297 mmol) in THF (5 mL) was added 10 drops of H2SO4 solution, and the reaction was stirred at room temperature for 1 h. After completion of the reaction, the reaction mixture was poured into 10 mL of NaHCO3 solution and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the orange-red solid of compound 10-4 (392 mg, crude) was obtained. The crude product was directly used in the next step. LCMS (ESI, m / z): 541.3 [M+H] + .

[0182] Fourth step: The crude compound 10-4 (392 mg) was dissolved in trifluoroacetic acid (4 mL) and stirred at room temperature for 1 h. The reaction mixture was concentrated to give 400 mg of crude product. The crude product was dissolved in ammonia in methanol solution (7 M, 4 mL) and stirred at room temperature for 1 h. After completion of the reaction, the product was dissolved in DMF and purified by reverse phase column to give compound 10. LCMS (ESI, m / z): 411.1 [M+H] + . 1H NMR (400 MHz, DMSO-d6) δ 11.92 (s, 1H), 7.93 (s, 1H), 7.85 (s, 1H), 7.79 (d, J = 7.9 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.62-7.55 (m, 2H), 7.04 (s, 2H), 6.21 (s, 1H), 3.91-3.80 (m, 1H), 3.70 (s, 3H), 3.52-3.38 (m, 2H), 2.80-2.64 (m, 2H), 2.02 (dq, J = 12.4, 8.5 Hz, 1H), 1.51-1.41 (m, 1H), 1.36-1.23 (m, 1H), 1.10-0.97 (m, 1H).

[0183] Example 11: Synthesis of (R)-3-(6-(l-methyl-lH-pyrazol-4-yl)-3,6,8a,9,10,11- hexahydro-8H-pyrrolo[2,l-c]pyrrolo[3',2':5,6]pyrido[4,3-e][l,4]oxazepin-l- yl)benzonitrile (Compound 11)

[0184]

[0185] Compound 11 was prepared according to the procedure described in Example 10, except that enantiomeric compound 2-2 was used in place of compound 1-2 in the first step. LCMS (ESI, m / z): 411.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.92 (s, 1H), 7.93 (s, 1H), 7.85 (s, 1H), 7.79 (d, J = 7.9 Hz, 1H), 7.71 (d, J = 7.8 Hz, 1H), 7.62-7.55 (m, 2H), 7.04 (s, 2H), 6.21 (s, 1H), 3.91-3.80 (m, 1H), 3.70 (s, 3H), 3.52-3.38 (m, 2H), 2.80-2.64 (m, 2H), 2.02 (dq, J = 12.4, 8.5 Hz, 1H), 1.51-1.41 (m, 1H), 1.36-1.23 (m, 1H), 1.10-0.97 (m, 1H).

[0186] Example 12: Synthesis of (S)-3-(8-oxo-3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,l- c]pyrrolo[3',2':5,6]pyrido[4,3-e][l,4]oxazepin-l-yl)benzonitrile (Compound 12)

[0187]

[0188] First Step: To a solution of compound INT1 (2 g, 5.1 mmol) in DMSO / isopropanol (10 / 1, 22 mL), add L-proline methyl ester (3.3 g, 25.7 mmol) and DI petroleum ether A (3.3 g, 25.7 mmol), stir the reaction at 110 °C for 1 h. After the reaction is completed, pour the reaction into 50 mL water and extract with ethyl acetate (50 mL x 2), wash the organic phase with saturated sodium chloride (30 mL x 3) and dry over anhydrous sodium sulfate, filter, concentrate, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate; ethyl acetate: 0%~20%), to obtain compound 12-2 (680 mg, 27.5%) as a yellow oil. LCMS (ESI, m / z): 481.7 [M+H] + .

[0189] Second Step: To a solution of compound 12-2 (1 g, 2.1 mmol) in ethanol (20 mL), add NaBH4 (86 mg, 2.3 mmol) at 0 °C, stir the reaction at 0 °C for 1 h. Slowly pour the reaction into 50 mL saturated ammonium chloride and extract with DCM (50 mL x 2), wash the organic phase with saturated sodium chloride (50 mL x 2) and dry over anhydrous sodium sulfate, filter, concentrate, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate; ethyl acetate: 0%~70%), to obtain compound 12-3 (230 mg, 22.9%) as a yellow liquid. LCMS (ESI, m / z): 483.8 [M+H] + .

[0190] Third Step: To a solution of compound 12-3 (210 mg, 433.5 umol) in dioxane / water (10 / 1, 5 mL), add 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (129 mg, 563.5 umol), Pd(dtbpf)Cl2 (28 mg, 43.4 umol) and potassium phosphate (276 mg, 1.3 mmol), stir the reaction at 80 °C for 3 h under nitrogen protection. After the reaction is completed, concentrate the reaction to dryness, and purify the crude product by silica gel column chromatography (petroleum ether / ethyl acetate; ethyl acetate: 0%~50%), to obtain compound 12-4 (190 mg, crude) as a light yellow oil. LCMS (ESI, m / z): 375.1 [M+H] + .

[0191] Fourth step: To a mixture of compound 12-4 (130 mg, 347.2 umol) in EtOH / tetrahydrofuran (1 / 1, 6 mL) was added sodium acetate (142 mg, 1.7 mmol) at room temperature. The reaction was stirred at room temperature for 5 hours. After completion, it was concentrated and the crude was purified by preparative high performance liquid chromatography to give compound 12. LCMS (ESI, m / z): 345.1 [M+H] + . 1 H NMR (400 MHz, CDC13) δ 8.00 (s, 1H), 7.79 (s, 1H), 7.74 (d, J = 7.8 Hz, 1H), 7.60 (d, J = 7.7 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.32 (s, 1H), 5.83 (d, J = 14.3 Hz, 1H), 5.17 (d, J = 6.3 Hz, 1H), 5.01 (d, J = 14.4 Hz, 1H), 3.06 (td, J = 9.6, 4.4 Hz, 1H), 2.71 (dd, J = 17.2, 9.1 Hz, 1H), 2.54 (t, J = 9.9 Hz, 1H), 2.08 - 2.06 (m, 1H), 1.70 (ddd, J = 17.0, 8.3, 4.2 Hz, 1H), 1.32 - 1.23 (m, 1H).

[0192] Example 13: Synthesis of (S)-3-(6-oxo-3,6,8a,9,10,11-hexahydro-8H-pyrrolo[2,1- c]pyrrolo[3',2':5,6]pyrido[4,3-e][l,4]oxazocin-l-yl)benzonitrile (Compound 13)

[0193]

[0194] First step: To a solution of compound 13-1 (8.6 g, 40.76 mmol) in DMF (100 mL) was added NaH (2.45 g, 61.14 mmol) at 0 °C, the reaction was stirred at 0 °C for half an hour, then 2-(trimethylsilyl)ethoxymethyl chloride (10.19 g, 61.14 mmol) was added, the reaction was stirred at room temperature for 2 hours, after completion, 60 mL saturated ammonium chloride solution and 200 mL water were added, extracted with ethyl acetate (200 mL x 3), the organic phase was washed with saturated brine (60 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, the crude was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 4 / 1) to give compound 13-2 (3.92 g, 28%) as yellow oil. LCMS (ESI, m / z): 341.0 [M+H] + .

[0195] Step 2: To a solution of compound 13-2 (3.8 g, 11.14 mmol) in DMF (50 mL) was added compound N-bromosuccinimide (2.18 g, 12.25 mmol). The reaction was stirred at room temperature for 2 hours. After completion, the reaction was diluted with water (150 mL) and extracted with ethyl acetate (70 mL x 3). The organic phase was washed with saturated brine (40 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 9 / 1) to obtain compound 13-3 (1.72 g, 36.8%) as a light yellow solid. LCMS (ESI, m / z): 421.1 [M+H] + .

[0196] Step 3: To a solution of compound 13-3 (1.2 g, 2.857 mmol) in isopropanol (15 mL) were added (S)-pyrrolidin-2-ylmethanol (433 mg, 4.286 mmol) and DI petroleum ether A (737 mg, 5.714 mmol). The reaction was stirred at 70°C for 3 hours. After completion of the reaction, the reaction solution was concentrated and the crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 85 / 15) to obtain compound 13-4 (950 mg, 68.7%) as a yellow oil. LCMS (ESI, m / z): 483.9 [M+H] + .

[0197] Step 4: To a solution of compound 13-4 (830 mg, 1.71 mmol) in methanol (10 mL) and water (2 mL) was added LiOH (415 mg, 17.29 mmol) and stirred at room temperature for 2 hours. After the reaction was complete, water (15 mL) was added for dilution, and the pH was adjusted to 4-5 with dilute hydrochloric acid. The mixture was extracted with ethyl acetate (10 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 13-5 (720 mg, 89.7%) as a light yellow oil. LCMS (ESI, m / z): 469.9 [M+H] + .

[0198] Step 5: To a solution of compound 13-5 (720 mg, 1.53 mmol) in dichloromethane (10 mL) were added DCC (1.9 g, 9.20 mmol) and DMAP (19 mg, 0.15 mmol). The reaction was stirred at 0°C under nitrogen for 2 hours. After completion of the reaction, the mixture was concentrated and the crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 20:1) to afford compound 13-6 (620 mg, 89.8%) as a pale yellow oil. LCMS (ESI, m / z): 454.0 [M+H] + .

[0199] Sixth step: To a solution of compound 13-6 (200 mg, 0.44 mmol) in 1,4-dioxane (2 mL) and water (0.2 mL), 3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)benzonitrile (153 mg, 0.66 mmol), potassium phosphate (189 mg, 0.89 mmol) and Pd(dtbpf)Cl2(30 mg, 0.04 mmol) were added, and the reaction was stirred at 70 °C for 2 h under nitrogen. After the reaction was completed, it was concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 6: 1) to obtain compound 13-7 (220 mg, 84.3%) as a light yellow solid. LCMS (ESI, m / z): 475.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.65 (s, 1H), 8.04 (s, 1H), 7.93-7.89 (m, 2H), 7.85-7.81 (m, 1H), 7.69 (t, J = 7.8 Hz, 1H), 5.68 (s, 2H), 4.70 (d, J = 12.3 Hz, 1H), 4.60 (dd, J = 13.1, 4.1 Hz, 1H), 3.88-3.82 (m, 1H), 3.66 (t, J = 10.6, 5.4 Hz, 2H), 2.98-2.91 (m, 1H), 2.67-2.62 (m, 1H), 1.83-1.57 (m, 4H), 0.92 (t, 2H), -0.00 (s, 9H).

[0200] Seventh step: To a solution of compound 13-7 (200 mg, 0.42 mmol) in dichloromethane (1 mL), trifluoroacetic acid (1.5 mL) was added, and the reaction was stirred at room temperature for 1 h. After the reaction was completed, it was concentrated, and the crude product was added to an ammonium methanol solution (7 M, 2 mL) and stirred at room temperature for 1 h. After the reaction was completed, it was concentrated, and the crude product was purified by preparative high performance liquid chromatography to obtain compound 13. LCMS (ESI, m / z): 345.0 [M+H] + . 1H NMR (400 MHz, DMSO) δ 12.31 (s, 1H), 8.55 (s, 1H), 7.98 (s, 1H), 7.85 (d, J = 7.9 Hz, 1H), 7.74 (d, J = 7.7 Hz, 1H), 7.67 (d, J = 2.5 Hz, 1H), 7.60 (t, J = 7.8 Hz, 1H), 4.63 (d, J = 13.0 Hz, 1H), 4.54 (dd, J = 13.0, 3.9 Hz, 1H), 3.81 (s, 1H), 2.94 - 2.86 (m, 1H), 2.66 - 2.60 (m, 1H), 2.05 - 1.94 (m, 1H), 1.80 - 1.67 (m, 1H), 1.45 - 1.30 (m, 2H).

[0201] Example 14: Synthesis of (S)-3-(7a,8,9,10-tetrahydro-3H,7H-pyrrolo[l,2-d]pyrrolo[3',2':5,6]pyrido[3,4-b][l,4]oxazin-l- yl)benzonitrile (Compound 14)

[0202]

[0203] First Step: To a solution of compound 14-1 (5.03 g, 21.6 mmol) in tetrahydrofuran (50 mL) was added NaH (1.03 g, 25.9 mmol) in an ice-water bath, after the reaction was stirred for 30 min, 2-(trimethylsilyl)ethoxymethyl chloride (4.32 g, 25.9 mmol) was added, the reaction was stirred at room temperature for 1 h. After the reaction was completed, the reaction system was slowly poured into saturated ammonium chloride solution (90 mL), extracted with ethyl acetate (90 mL x 3), washed with saturated brine (90 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to give colorless liquid compound 14-2 (5.64 g, yield 72.18%). LCMS (ESI, m / z): 363.9 [M+H] + .

[0204] Second Step: To a solution of compound 14-2 (1 g, 2.764 mmol) in acetonitrile (10 mL) was added N-chlorosuccinimide (554 mg, 4.147 mmol), the reaction was stirred at room temperature for 5 hours. After the reaction was completed, the reaction system was slowly poured into saturated NaHCO3(40 mL), extracted with ethyl acetate (40 mL x 3), washed with saturated brine (40 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to give compound 14-3 (884 mg, yield 80.7%) as a light yellow solid. LCMS (ESI, m / z): 396.9 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.57 (s, 1H), 8.07 (s, 1H), 5.59 (s, 2H), 3.51 (s, 2H), 0.83-0.79 (m, 2H), -0.10 (s, 9H).

[0205] Third Step: To a solution of compound 14-3 (835 mg, 2.11 mmol) in NMP (8 mL) was added (S)-pyrrolidin-2-ylmethanol (426.4 mg, 4.22 mmol), the reaction was stirred at 180°C for 4 hours. After the reaction was completed, diluted with water (30 mL), extracted with ethyl acetate (30 mL x 3), washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to give compound 14-4 (550 mg, yield 56.6%) as a light yellow oil. LCMS (ESI, m / z): 462.0 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 8.49 (s, 1H), 7.92 (s, 1H), 5.69-5.60 (m, 2H), 4.50 (t, J = 5.4 Hz, 1H), 4.19-4.08 (m, 1H), 3.63 (ddd, J = 13.0, 7.8, 3.5 Hz, 3H), 3.32-3.19 (m, 3H), 2.28 (dq, J = 11.8, 8.2 Hz, 1H), 2.07 (dt, J = 7.8, 6.1 Hz, 2H), 1.97 (ddd, J = 11.1, 7.0, 3.6 Hz, 1H), 0.93-0.86 (m, 2H), 0.02--0.03 (m, 9H).

[0206] Fourth step: To a solution of compound 14-4 (500 mg, 1.1 mmol) in 1,4-dioxane (10 mL), CuI (41 mg, 0.217 mmol), N,N'-dimethyl-1,2-cyclohexanediamine (62 mg, 0.434 mmol) and potassium phosphate (691 g, 3.3 mmol) were added. The reaction was stirred at 110 °C for 12 h under nitrogen protection. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) to give compound 14-5 (316 mg, yield 75.8%) as a light yellow oil. LCMS (ESI, m / z): 380.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) d 7.81 (s, 1H), 7.59 (s, 1H), 5.48 (dd, J = 23.1, 10.7 Hz, 2H), 4.22 (dd, J = 10.3, 3.3 Hz, 1H), 4.10 (dt, J = 10.1, 7.1 Hz, 1H), 3.51 - 3.44 (m, 4H), 3.32 (s, 1H), 2.16 (ddd, J = 16.4, 10.3, 6.3 Hz, 1H), 1.90 - 1.77 (m, 2H), 1.76 - 1.68 (m, 1H), 0.83 - 0.76 (m, 2H), -0.08 - -0.11 (m, 9H).

[0207] Fifth step: To a solution of compound 14-5 (296 mg, 0.703 mmol) in 1,4-dioxane / water (10 / 1, 3 mL), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzonitrile (242 mg, 1.054 mmol), Pd(dtbpf)Cl2 (46 mg, 0.0703 mmol) and potassium carbonate (291 mg, 2.108 mmol) were added. The reaction was stirred at 90 °C for 3 h under nitrogen protection. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic layers were washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to give compound 14-6 (171 mg, yield 54.5%) as a white solid. LCMS (ESI, m / z): 447.1 [M+H] + .

[0208] Step 6: Compound 14-6 (171 mg, 0.383 mmol) was dissolved in trifluoroacetic acid (3 mL) and stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated twice with acetonitrile to obtain the crude product. LCMS (ESI, m / z): 347.0 [M+H] + .

[0209] Compound 14 was obtained by dissolving the above-mentioned crude product in NH3-MeOH (7 M, 3 mL) solution and stirring at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by preparative high performance liquid chromatography. LCMS (ESI, m / z): 317.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ 11.88 (s, 1H), 8.03-7.92 (m, 2H), 7.87 (d, J = 7.9 Hz, 1H), 7.70 (d, J = 7.7 Hz, 1H), 7.62 (s, 1H), 7.57 (t, J = 7.8 Hz, 1H), 4.78 (s, 2H), 4.14-3.88 (m, 2H), 3.55-3.44 (m, 2H), 2.29 (s, 3H).

[0210] Biological evaluation

[0211] Test Example 1: LRRK2 wt kinase inhibition activity assay

[0212] DNL201 was synthesized according to the compound 410 on page 184 of the specification of WO2012062783A1.

[0213] Purpose of the experiment: To detect the energy transfer (520nM / 485nM fluorescence signal ratio) generated after the phosphoric group of phosphorized Fluorescein-ERM (LRRKtide) peptide and Tb-ERM (LRRKtide) Antibody binding by time-resolved fluorescence resonance energy transfer technology (TR-FRET). The IC 50 value of the test compound on LRRK2 kinase was calculated.

[0214] Principle of the experiment: This protocol describes an in vitro method for measuring the phosphorylation of a peptide substrate by WT-LRRK2 enzyme. The kinase reaction is performed by incubating a fluorescein-labeled substrate with recombinant human kinase and ATP. The Lantha Screen TM Kinase activity is determined by LanthaScreen TMDuring the kinase reaction, a terbium-labeled antibody (used to detect phosphorylated products) binds to a phosphorylated fluorescein-labeled substrate, resulting in an increase in the TR-FRET value. The TR-FRET value is determined by the ratio of the FRET-specific signal measured with a 520nm filter to the signal measured with a 495nm filter. The amount of antibody bound to the tracer is proportional to the amount of phosphorylated substrate present. Therefore, the increase in the TR-FRET value can be used to measure kinase activity.

[0215] Material:

[0216] Reagents:

[0217]

[0218] Consumables:

[0219]

[0220] instrument:

[0221]

[0222]

[0223] Experimental steps:

[0224] 1. Add the DMSO solution of the test compound to a 384-well microplate using the Echo655 non-contact nano-ultrasonic dispensing system; the highest concentration is 10uM, with a 3-fold gradient dilution and 11 concentration gradients.

[0225] 2. Prepare a mixed enzyme and peptide solution using freshly prepared reaction solution (final concentration: 2 nM enzyme + 0.4 uM Fluorescein-ERM (LRRKtide) petroleum ether ptide). Add 5 uL to the 384-well microplate containing the compound and centrifuge at 1000 rpm / min for 1 minute. Incubate at room temperature for 15 minutes.

[0226] 3. Add 5 μL ATP (final concentration: 38 μM) and centrifuge at 1000 rpm / min for 1 minute. Incubate at room temperature for 120 minutes.

[0227] 4. Add 10uL of detection reagent: Tb-petroleum ether RM(pLRRKtide) Antibody (final concentration: 0.25 nM) and EDTA (final concentration: 10 mM) were centrifuged at 1000 rpm / min for 1 min. The reaction was allowed to proceed at room temperature for 30 min.

[0228] 5. Envison detects TR-TRET fluorescence signal, mirror 447 (D400 / D505), filter 275 (520 nm) and 102 (485 nm);

[0229] 6. Calculate the inhibition of enzyme activity by the compound by signal ratio (520 nm / 485 nm), and use software XLfit 5 to fit the curve to calculate IC 50 value.

[0230] Quality control: if the Z value is >0.5, the IC 50 of the reference compound LRRK2-IN-1 is consistent with historical data (2-17 nM), then the data QC is passed.

[0231] Experimental results:

[0232] Compound number <![CDATA[LRRK2,IC 50 (nM)]]> Compound number <![CDATA[LRRK2,IC 50 (nM)]]> DNL201(Yang Shen) 7.39 Compound 8 1.10 Compound 1 0.379 Compound 9 9.38 Compound 2 1.37 Compound 10 6.88 Compound 3 1.09 Compound 11 15.7 Compound 4 0.755 Compound 12 97.5 Compound 5 0.395 Compound 13 2.40 Compound 6 0.453 Compound 14 1.53 Compound 7 1.30

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in: X is O, S, NH or CH2; Y is a single bond, C(=O) or CHR 4 ; R 1 is optionally replaced by 1 or 2 R 1a a substituted aryl or heteroaryl group, wherein the heteroaryl group contains at least one nitrogen atom; R 1a It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 2 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 3 It is hydrogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy or oxo; R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom; n is 0, 1, or 2; m is 0 or 1.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein: X is O or CH2; R 1 is phenyl, pyrrolyl or pyridyl; R 1a It is hydrogen, C 1-6 Alkyl, cyano or halogen; R 2 It is hydrogen, C 1-6 Alkyl or halogen; R 3 is hydrogen or oxo; R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl or optionally C 1-6 Alkyl-substituted pyrazolyl.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein: R 1 is phenyl or pyrrolyl; R 1a is hydrogen, methyl or cyano; R 2 is hydrogen or a halogen; R 4 is hydrogen, methyl, trifluoromethyl or pyrazolyl optionally substituted with methyl.

4. The compound according to claim 1 or 3, which has the specified stereochemical formula (I'):

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a chemical structure of formula (II), in: Y is a single bond, C(=O) or CHR 4 ; R 1 is optionally replaced by 1 or 2 R 1a a substituted aryl or heteroaryl group, wherein the heteroaryl group contains at least one nitrogen atom; R 1a It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 2 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 3 It is hydrogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy or oxo; R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom; n is 0, 1, or 2.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a chemical structure of formula (III), in: R 1 is optionally replaced by 1 or 2 R 1a a substituted aryl or heteroaryl group, wherein the heteroaryl group contains at least one nitrogen atom; R 1a It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 2 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 alkylamino, hydroxy, amino, cyano or halogen; R 3 It is hydrogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy or oxo; R 4 It is hydrogen, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, hydroxy or optionally C 1-6 an alkyl-substituted 5-6 membered heteroaryl group containing at least one nitrogen atom; n is 0, 1, or 2.

7. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a chemical structure of formula (IIa) or formula (IIb), in: R 1 is optionally replaced by 1 or 2 R 1a substituted phenyl or pyrrolyl; R 1a It is hydrogen, C 1-6 Alkyl or cyano; R 4 It is hydrogen, C 1-6 Alkyl or C 1-6 Halogenated alkyl.

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein: R 1 yes R 4 is hydrogen, methyl or trifluoromethyl; preferably, R 4 It's hydrogen.

9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

10. A pharmaceutical composition comprising the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

11. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 8, in the preparation of a medicament for treating LRRK2 receptor-related disorders.

12. The use according to claim 9, wherein the disease is a neurodegenerative disease, an immune-inflammatory disease, a tumor or glaucoma.

13. The method according to claim 10, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, Huntington's disease, spinocerebellar ataxia, Friedreich's ataxia, Pick's disease, dementia with Lewy bodies, dystonia, amyotrophic lateral sclerosis, neuroinflammation, progressive supranuclear palsy and frontotemporal dementia.

14. The method according to claim 10, wherein the immune inflammation-related disease is selected from inflammatory bowel disease, systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, ankylosing spondylitis, autoimmune hemolytic anemia, pure red cell aplasia, idiopathic thrombocytopenic purpura, Evan's syndrome, vasculitis, bullous skin disorders, type I diabetes, Sjögren's syndrome, Devic's disease and inflammatory myopathy.

Images