SARM1 enzyme activity inhibitor and use thereof

Abstract

Provided in the present invention is the use of an SARM1 enzyme activity inhibitor in treatment of neurodegenerative diseases or neurological diseases or disorders. Particularly provided in the present invention are compounds of formula (I) serving as SARM1 enzyme activity inhibitors and a pharmaceutical composition thereof.

Description

SARM1 enzyme activity inhibitors and uses thereof Technical Field

[0001] The present application relates to compounds that can be used to inhibit SARM1 enzyme activity, and / or the use of these compounds in treating and / or preventing neurodegenerative or neurological diseases or conditions associated with SARM1 enzyme activity. Background Art

[0002] Neurodegenerative diseases are a class of diseases that can severely harm humans, causing devastating damage, such as progressive neuronal cell death. Among the primary neurodegenerative diseases, known are central nervous system diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), and Huntington's disease, as well as peripheral nervous system diseases such as diabetes. While most are associated with aging, the incidence of these diseases increases with age, although some also occur in middle-aged individuals or even younger.

[0003] As a result of research on brain structure and function, the effects of neurotransmitters and neurotrophic factors have been gradually elucidated, but many local causes of neurodegeneration are still unclear. Only for Parkinson's disease, the relationship between this disease and the special neurotransmitter, i.e., dopamine, has been elucidated, and the precursor of dopamine, L-dopa, has been used as a medicine to alleviate neurological symptoms and restore neurological function. However, L-dopa cannot inhibit the development of neurodegeneration, and gradually loses its effect as the disease progresses, i.e., dopamine-based neuronal cell degeneration and defect. Similarly, Alzheimer's disease is also caused by the degeneration and defect of multiple nerve cells such as acetylcholine-based neurons, monoamine-based neurons, etc., and as medicines for the treatment of this disease, cholinesterase inhibitors have been put on the market or are being developed. However, the L-dopa for the treatment of Parkinson's disease is still limited to symptomatic treatment, to temporarily improve neurological symptoms.

[0004] Therefore, there is a lack of effective therapeutic drugs for neurodegenerative diseases.

[0005] Studies have found that axonal damage occurs in a variety of neurodegenerative diseases, accidental injuries, and other neurological diseases. Axonal degeneration can cause structural necrosis and functional disorders in the peripheral nervous system, ultimately leading to acquired or hereditary degenerative diseases of the central nervous system.

[0006] Although there is currently no very effective pharmacological method that can accurately assess the weight of the morbidity caused by axonal degeneration, it has been found in histopathological studies that significant axonal damage degradation has been observed in the early stages of various neuropathies such as Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, and peripheral neuropathy, indicating that axonal degeneration plays an important role in the development of neuropathy (Fischer et al., Neuro-degenerative Diseases, 2007, 4:431-442). Therefore, by weakening or even blocking axonal degeneration, maintaining the integrity of neuronal structure and function may be a therapeutic solution that benefits various nervous system diseases.

[0007] In the absence of effective therapeutic drugs for neurodegenerative diseases, the existing technology urgently needs to research and develop new compounds, especially small chemical molecules, including compounds that have effects on axonal degeneration.

[0008] Summary of the Invention

[0009] After long-term research, the present inventors unexpectedly discovered a class of compounds with significant SARM1 enzyme activity inhibitory effects, and found that the compounds can improve axonal degeneration and be used to treat or prevent neurodegenerative diseases and related disorders.

[0010] The SARM1 (Sterile alpha and TIR motif containing 1) protein is composed of three domains: the ARM (Armadillo / HEAT repeat) domain at the nitrogen terminus, two tandem SAM (Sterile alpha motif) domains, and the TIR (Toll / Interleukin Receptor) domain at the carbon terminus. In addition, there is a mitochondrial localization signal peptide at the nitrogen terminus.

[0011] It is known that in wild-type neurons, axonal injury induces NAD + Depletion and axonal degeneration; SARM1 knockout inhibits axonal degeneration, and NAD + Maintained at normal levels, indicating that SARM1 promotes NAD + consumption, exacerbating axonal degeneration.

[0012] The Milbrandt team at the University of Washington School of Medicine prepared the TIR domain of SARM1 (SARM1-TIR) and found that it has NAD + Hydrolase activity. Further rigorous E. coli expression and purification experiments and cell-free expression systems were used to obtain high-purity SARM1-TIR, ultimately proving that SARM1-TIR can catalyze NAD + Produces adenosine 5'-diphosphate ribose (ADPR) and cyclic adenosine 5'-diphosphate ribose (cADPR).

[0013] SARM1 is a multifunctional signaling enzyme that catalyzes the conversion of multiple substrates to NAD + , NADP + and NA to generate signal molecules cADPR, ADPR and NAADP. In a variety of neurodegenerative diseases, SARM1 is activated, leading to NAD + Depletion of SARM1 can activate a new cell death mechanism; knocking out SARM1 can inhibit axonal degeneration and disease progression, and is therefore considered a potential drug target for related neurological diseases, including TBI, AD, CIPN, ASL, etc.

[0014] In the present disclosure, the inventors prepared full-length SARM1 for NAD enzyme activity experiments, and used it to screen and obtain compound molecules with enzyme activity inhibitory ability of the present invention.

[0015] Therefore, based on the above findings, in a first aspect, the present invention provides use of a SARM1 enzyme activity inhibitor in the preparation of a medicament for treating or preventing neurodegenerative diseases or neurological diseases or disorders.

[0016] In another aspect, the present invention provides use of a SARM1 enzyme activity inhibitor in the preparation of a medicament for treating or preventing axonal degeneration-related diseases or disorders.

[0017] In particular, the present invention provides compounds of formula (I) that are useful as inhibitors of SARM1 enzyme activity:

[0018] or its racemate, enantiomer, diastereomer, deuterated form or pharmaceutically acceptable salt, wherein,

[0019] X1, X2, X3, X4, X5 are each independently selected from CH or N;

[0020] Z represents -CH2-, -CH2(CH3)- or -O-;

[0021] represents a single bond or a double bond;

[0022] Y represents a carbon atom or a nitrogen atom;

[0023] R1 is independently selected from H, halogen, C1-C3 alkyl, cyano, trifluoromethyl, amino, hydroxy, methoxy-C(O)NH2, preferably H, F, Cl, methyl, cyano and -C(O)NH2;

[0024] R2 is independently selected from H, halogen, C1-C3 alkyl, cyano, trifluoromethyl, amino, hydroxy, methoxy, C1-C3 alkyl-OC(O)-, C1-C3 alkyl-NHC(O)-, preferably H, cyano, Cl, methyl and methoxy;

[0025] M stands for O or S;

[0026] L is selected from -CH2-CH2-, -CH2-, -O- and -CO-, wherein one or two -CH2- may be optionally replaced by -O-, -S-, -CO- or -NH-; preferably, L is selected from -CH2-, -CH2-CH2-, -CH-O-, -O- and -CO-;

[0027] E is selected from

[0028] Wherein, X6, X7, X8 are each independently selected from CH or N;

[0029] R3 is independently selected from H, halogen, methyl, cyano, trifluoromethyl, nitro, preferably H, F, Cl, Br, cyano, trifluoromethyl, nitro;

[0030] R4 is independently selected from H, halogen, methyl, cyano, trifluoromethyl, and nitro, preferably H, F, Cl, Br, cyano, trifluoromethyl, and nitro.

[0031] In some embodiments, R3 and R4 are each independently selected from H, halogen, methyl, cyano, trifluoromethyl, nitro, acetyl, methoxycarbonyl, carboxyl, ethynyl, ClCH=CH- and

[0032] In some preferred embodiments, E is selected from wherein X6, X7, X8, R3 and R4 have the same meanings as above.

[0033] In some preferred embodiments, the compound of formula (I) has the following structure:

[0034] wherein X1, X2, X3, X4, X5, L, Y, R1, R3 and R4 have the same meanings as above.

[0035] In some preferred embodiments, the compound of the present invention is selected from the following compounds, or racemates, enantiomers, diastereomers, deuterated compounds or pharmaceutically acceptable salts thereof:

[0036] Herein, when referring to a compound of formula (I) or formula (II), it also includes the pharmaceutically acceptable salts of the compound of formula (I) or formula (II) or stereoisomers thereof.

[0037] The present invention also relates to a method for treating or preventing a neurodegenerative disease or a neurological disease or condition related thereto, comprising administering to a subject in need thereof a compound of the present invention that is an inhibitor of SARM1 enzyme activity. In particular, the present invention relates to a method for treating or preventing a disease or condition related to axonal degeneration, comprising administering to a subject in need thereof a compound of the present invention that is an inhibitor of SARM1 enzyme activity. More particularly, the present invention relates to a method for inhibiting SARM1 enzyme activity, comprising administering to a subject in need thereof a compound of the present invention; more particularly, the present invention relates to a method for inhibiting axonal degeneration, comprising administering to a subject in need thereof a compound of the present invention. The compounds or compositions of the present invention can be administered to a subject or patient in need thereof in an effective amount.

[0038] Accordingly, the present invention also relates to the use of the compounds of the present invention or their pharmaceutically acceptable salts or stereoisomers in the preparation of a method for treating or preventing a neurodegenerative disease or a neurological disease or condition. The present invention also relates to the use of the compounds of the present invention or their pharmaceutically acceptable salts or stereoisomers in the preparation of a SARM1 enzyme activity inhibitor. The present invention also relates to the use of the compounds of the present invention or their pharmaceutically acceptable salts or stereoisomers in the preparation of a method for treating or preventing axonal degeneration-related diseases or conditions. Preferably, the neurodegenerative disease or neurological disease or condition or axonal degeneration-related disease or condition is selected from Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, and peripheral neuropathy. DETAILED DESCRIPTION

[0039] the term

[0040]

[0046] Reference herein to a "compound" having a particular structural formula is generally also intended to encompass pharmaceutically acceptable salts, stereoisomers, diastereomers, enantiomers, racemic mixtures, and isotopic derivatives thereof.

[0041] Pharmaceutically acceptable salts of the present invention may be formed using, for example, the following inorganic or organic acids: "Pharmaceutically acceptable salts" refers to salts that, within the scope of sound medical judgment, are suitable for use in contact with human and mammalian tissues without undue toxicity, irritation, allergic reaction, or the like, at a reasonable benefit / risk ratio. The salts may be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base or free acid with a suitable reagent. For example, the free base may be reacted with a suitable acid. Examples of pharmaceutically acceptable acid addition salts are salts of an amino group (amine group) formed with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or organic acids (e.g., acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or salts formed using other methods known in the art, such as ion exchange. Other pharmaceutically acceptable salts include sodium alginate, ascorbate, benzenesulfonate, adipate, camphorsulfonate, aspartate, benzoate, bisulfate, borate, butyrate, camphorate, citrate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, heptanoate, hexanoate, hydroiodide, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like.

[0042] The pharmaceutically acceptable salts of the present invention can be prepared by conventional methods, for example, by dissolving the compound of the present invention in a water-miscible organic solvent (e.g., methanol, ethanol, acetone and acetonitrile), adding an excess of an organic acid or an aqueous inorganic acid solution thereto to precipitate the salt from the resulting mixture, removing the solvent and the remaining free acid therefrom, and then isolating the precipitated salt.

[0043] The "stereoisomerism" mentioned in the present invention is divided into conformational isomerism and configurational isomerism. Configurational isomerism can be further divided into cis-trans isomerism and optical isomerism (i.e., optical isomerism). Conformational isomerism refers to a stereoisomerism phenomenon in which the atoms or atomic groups of an organic molecule with a certain configuration have different spatial arrangements due to the rotation or distortion of carbon-carbon single bonds. Common examples include the structures of alkanes and cycloalkanes, such as the chair conformation and boat conformation that appear in the structure of cyclohexane. "Stereoisomers" refer to when the compounds of the present invention contain one or more asymmetric centers, and can thus be racemates and racemic mixtures, single enantiomers, diastereomeric mixtures, and single diastereoisomers. The compounds of the present invention have asymmetric centers, and each asymmetric center will produce two optical isomers. The scope of the present invention includes all possible optical isomers and diastereomeric mixtures and pure or partially pure compounds.

[0044] In some cases, the compounds of the present invention can exist as tautomers, which have different hydrogen attachment points through one or more double bond displacements. For example, a ketone and its enol form are keto-enol tautomers. Amides and imines can also form tautomeric forms. Each tautomer and mixture thereof are included in the compounds of the present invention. Enantiomers, diastereomers, racemates, mesomorphs, cis-trans isomers, tautomers and mixtures thereof of all compounds are included within the scope of the present invention.

[0045] When the compound of the present invention is used in combination with another SARM1 enzyme activity inhibitor for the treatment or prevention of neurodegenerative diseases or related neurological diseases or conditions, or can be used in combination with another active drug for the treatment or prevention of neurodegenerative diseases or related neurological diseases or conditions, it is used to treat or prevent neurodegenerative diseases or related diseases or conditions.

[0046] In this article, the wavy line Indicates the position at which a group or substituent is attached to the parent compound.

[0047] The compound of the present invention or its mesomer, racemate, enantiomer, diastereomer or its pharmaceutically acceptable salt can be used as an active ingredient by oral or parenteral administration, and the scope of its effective amount is 0.1 to 2000 mg / kg body weight / day, preferably 0.1 to 100 mg / kg body weight / day in the case of mammals including humans (weighing about 70 kg), and is administered every day in single or divided doses, or with or without a predetermined time. The dosage of the active ingredient can be adjusted according to multiple relevant factors (such as the situation of the subject to be treated, the type of disease and severity, the rate of administration and the doctor's opinion). In some cases, the amount less than the above dosage may be suitable.

[0048] The compound of the present invention or its mesoform, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt can be formulated into a pharmaceutical composition. The pharmaceutical composition can comprise the compound of the present invention or its mesoform, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

[0049] The pharmaceutical composition of the present invention can be formulated into dosage forms for oral administration or parenteral administration (including intramuscular, intravenous and subcutaneous routes, intratumor injection) according to any of the conventional methods, such as tablets, granules, powders, capsules, syrups, emulsions, microemulsions, solutions or suspensions.

[0050] The pharmaceutical composition of the present invention for oral administration can be prepared by mixing the active ingredient with a pharmaceutically acceptable carrier such as cellulose, calcium silicate, magnesium stearate, calcium stearate, corn starch, lactose, sucrose, dextrose, calcium phosphate, stearic acid, surfactants, suspending agents, gelatin, talc, emulsifiers and diluents. Examples of carriers employed in the injectable composition of the present invention are water, glycerides, saline solutions, alcohols, glycols, glucose solutions, ethers (e.g., polyethylene glycol 400), oils, fatty acids, fatty acid esters, surfactants, suspending agents and emulsifiers.

[0051] If not stated otherwise, conventional methods of mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used. In this application, if not stated otherwise, "or" or "and" may be used to mean "and / or".

[0052] In the specification and claims, a given chemical formula or name shall encompass all stereo and optical isomers and racemates in which such isomers exist. Unless otherwise indicated, all chiral (enantiomers and diastereomers) and racemic forms are within the scope of the present invention. Many geometric isomers of C=C double bonds, C=N double bonds, ring systems, etc. may exist in the compounds, and all such stable isomers are encompassed by the present invention.

[0053] The compounds of the present invention can be isolated in optically active or racemic forms. All methods for preparing the compounds of the present invention and the intermediates prepared therein are considered part of the present invention. When enantiomeric or diastereomeric products are prepared, they can be separated by conventional methods (e.g., by chromatography or fractional crystallization). It should be understood that all isomeric forms that may exist are included in the present invention.

[0054] Unless otherwise defined, when a substituent is noted as "optionally substituted," the substituent is selected, for example, from substituents such as alkyl, cycloalkyl, aryl, heterocyclyl, halogen, hydroxy, alkoxy, nitro, cyano, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, alkylthio, and the like.

[0055] The term "alkyl" or "alkylene" as used herein is intended to include both branched and straight chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms. 12 Alkyl, C1-C 10 C1-C8 alkyl, C1-C6 alkyl, more preferably C1-C4 alkyl, particularly preferably C1-C3 alkyl. For example, "C1-C6 alkyl" means an alkyl group having 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl) and pentyl (e.g., n-pentyl, isopentyl, neopentyl). For the C1-C6 alkyl group in the present invention, 12 In the case of an alkyl group, 1 to 4 -CH2- units are optionally replaced by O atoms, S atoms or -NH-.

[0056] The term "carbonyl" refers to an organic functional group composed of carbon and oxygen atoms connected by a double bond (C=O or C(O)).

[0057] The term "aromatic ring" refers to an aromatic ring structure. An aromatic carbocyclic ring refers to an aromatic carbocyclic ring structure in which all ring members are composed of carbon atoms. A non-aromatic carbocyclic ring refers to a non-aromatic carbocyclic ring structure in which all ring members are composed of carbon atoms.

[0058] "Halo" or "halogen" includes fluorine, chlorine, bromine and iodine; fluorine and chlorine are preferred.

[0059] The term "heterocycle" refers to a monocyclic or bicyclic heterocycloalkyl system, and may also include spiro heterocycles or bridged heterocycloalkyls. A monocyclic heterocycloalkyl system refers to a 3-8 membered, saturated or unsaturated, but non-aromatic, cyclic alkyl system containing at least one heteroatom selected from O, N, S, or P. A bicyclic heterocycloalkyl system refers to a heterocycloalkyl fused to a phenyl, a cycloalkyl, a cycloalkenyl, a heterocycloalkyl, or a heteroaryl group. An aromatic heterocycle refers to an aromatic heterocycle system, and a non-aromatic heterocycle refers to a non-aromatic heterocycle system.

[0060] When any variable occurs more than one time in any constituent or formula for a compound, its definition at each occurrence is independent of its definition at every other occurrence. Thus, for example, if a group is shown to be substituted with 0-3 R groups, then said group may be optionally substituted with up to three R groups, and at each occurrence R is independently selected from the definition of R. Furthermore, combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0061] As used herein, the term "effective amount" means an amount of a drug or pharmaceutical agent (i.e., a compound of the present invention) that will cause a biological or medical response in a tissue, system, animal, or human being, such as that sought by a researcher or clinician. In addition, the term "therapeutically effective amount" means an amount that results in improved treatment, cure, prevention, or alleviation of a disease, condition, or side effect, or reduces the rate of progression of a disease or condition, compared to a corresponding subject that has not received the above amount. An effective amount can be given in one or more administrations, applications, or dosages and is not intended to be limited by a specific formulation or route of administration. The term also includes within its scope an effective amount that enhances normal physiological function.

[0062] As used herein, the term "treating" includes any effect that results in improvement of a condition, disease, disorder, etc., such as alleviation, reduction, modulation, improvement, or elimination, or amelioration of the symptoms thereof.

[0063] The term "pharmaceutically acceptable" is used herein to refer to those compounds, substances, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response and / or other problems or complications, and commensurate with a reasonable benefit / risk ratio.

[0064] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharmaceutical substance, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, manufacturing aid (e.g., a lubricant, talc, magnesium stearate, calcium stearate, or zinc stearate, or stearic acid), or solvent encapsulating substance, which is involved in carrying or transporting the subject compound from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient.

[0065] The term "pharmaceutical composition" means a composition comprising a compound of the present invention and at least one other pharmaceutical carrier. "Pharmaceutical carrier" refers to a medium generally accepted in the art for delivering biologically active agents to animals (particularly mammals), including (i.e.) adjuvants, excipients or vehicles such as diluents, preservatives, fillers, flow regulators, disintegrants, wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, fragrances, antibacterial agents, antifungal agents, lubricants and dispersants, depending on the mode of administration and the nature of the dosage form.

[0066] The compounds or pharmaceutical compositions of the present invention can improve a disease, symptom, or condition after administration, particularly improving its severity, delaying its onset, slowing its progression, or reducing its duration, regardless of whether the administration is fixed or temporary, continuous or intermittent, and can be attributed to or related to the administration.

[0067] Route of administration

[0068] Suitable routes of administration for the compounds or pharmaceutical compositions of the present invention include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transdermal, vaginal, auricular, nasal, and topical administration. In addition, parenteral administration, by way of example only, includes intramuscular, subcutaneous, intravenous, intramedullary, intraventricular, intraperitoneal, intralymphatic, and intranasal injections.

[0069] In some embodiments, the administration of the compound of the present invention is topical administration. In other specific embodiments, the compound of the present invention can be administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. In addition, in other specific embodiments, the compound of the present invention is administered by a targeted drug delivery system.

[0070] In the pharmaceutical compositions of the present invention, pharmaceutical carriers can be formulated based on a variety of factors well within the purview of those skilled in the art. These factors include, but are not limited to: the type and nature of the active agent being formulated; the subject to whom the composition containing the active agent is to be administered; the intended route of administration of the composition; and the targeted therapeutic indication. Pharmaceutical carriers include aqueous and non-aqueous liquid media and various solid and semisolid dosage forms.

[0071] Such carriers may include a variety of ingredients and additives in addition to the active agent, which are included in the formulation for various reasons known to those skilled in the art, such as stabilizing the active agent, binding agents, etc. Descriptions of suitable pharmaceutical carriers and factors involved in carrier selection can be found in a number of readily available sources, such as Allen LV Jr. et al. Remington: The Science and Practice of Pharmacy (2 Volumes), 22 nd Edition(2012),Pharmaceutical Press.

[0072] The compounds are usually administered in admixture with suitable pharmaceutical diluents, excipients or carriers (collectively referred to herein as pharmaceutical carriers) appropriately selected according to the intended administration form (e.g., oral tablets, capsules, elixirs and syrups) and in accordance with conventional pharmaceutical practice.

[0073] While it is possible for a compound of the present invention to be administered alone, it is preferable to administer the compound as a pharmaceutical formulation (composition).

[0074] Kit / Product Packaging

[0075] Kits / product packaging are also described herein for use in treating the aforementioned indications. These kits can consist of a carrier, a pouch, or a container box, each of which can be divided into compartments to accommodate one or more containers, such as vials, test tubes, and the like, each containing a single component of the method. Suitable containers include bottles, vials, syringes, and test tubes. The containers can be made of acceptable materials such as glass or plastic.

[0076] For example, a container may contain one or more compounds described herein, either as a pharmaceutical composition or in admixture with other ingredients described herein. The container may have a sterile delivery port (e.g., an IV bag or bottle with a stopper pierceable by a hypodermic needle). Such a kit may include a compound and instructions, labeling, or operating instructions for use as described herein.

[0077] A typical kit may include one or more containers, each containing one or more materials (e.g., reagents, concentrated stock solutions, and / or instruments) to suit the commercialization and user needs of the compound. These materials include, but are not limited to, buffers, diluents, filters, needles, syringes, delivery devices, bags, containers, bottles, and / or test tubes, accompanied by a list of contents and / or instructions for use, including instructions for use within the packaging. The complete set of instructions should be included.

[0078] The features described above, or in the embodiments, may be combined in any combination. All features disclosed in this specification may be used in any combination, and each feature disclosed in this specification may be replaced by any alternative feature that serves the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the features disclosed are merely general examples of equivalent or similar features.

[0079] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight.

[0080] The units used in the present invention for weight-volume percentages are well known to those skilled in the art, for example, referring to the weight of solute in 100 ml of solution. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0081] In the terms used in the present invention, "neurodegenerative disease" and "neurodegenerative disease" have the same meaning; "axonal degeneration" and "axonal degeneration" have the same meaning. Those skilled in the art will understand that the terms have commonly understood meanings.

[0082] The descriptions provided herein, including those provided in the Examples, are not intended to limit the scope of any claims. The following non-limiting examples are provided to further illustrate the present invention. Based on this disclosure, those skilled in the art will appreciate that many changes may be made to the specific embodiments disclosed and still achieve the same or similar results without departing from the spirit and scope of the present teachings.

[0083] Unless otherwise noted, all materials / reagents were obtained from commercial suppliers and used without further purification. Reactions were monitored by LC-MS and / or thin-layer chromatography (TLC) on silica gel 60F254 (0.2 mm) pre-coated glass backing and observed using UV light. The structures of the following example compounds were characterized by nuclear magnetic resonance (NMR) and / or mass spectroscopy (MS).

[0084] 1 H NMR spectra (400 MHz) were recorded at room temperature on a Bruker spectrometer using TMS or residual solvent peaks as internal standards. Chemical shift values ​​or peak shape multiples are given in (δ), and coupling constants (J) are given as absolute values ​​in Hertz (Hz). Multiplicity in H NMR spectra is abbreviated as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br, or broad (broadened).

[0085] Preparative HPLC purification was performed on a Shimadzu LC-6AD. All purifications were performed using a Shim-pack PREP-DDS(H)KIT column. The mobile phase consisted of water (containing 0.1% HCO₂H₂) and acetonitrile; all reagents used were HPLC grade. The flow rate was 10 ml / min.

[0086] LC-MS was performed on an Agilent 1260 infinity II; mobile phase: A: water (0.1% trifluoroacetic acid), B: ACN; 3.5 min column run; column: YMC-Triart C18 50*3 mm, 3 um; flow rate: 1.8 ml / min; oven temperature: 40°C; gradient: 5-100 (ACN%).

[0087] Preparative TLC was performed on Whatman LK6F Silica Gel 60A plates of size 20 x 20 cm with a thickness of 500 μm.

[0088] The following examples are intended to illustrate embodiments of the present invention but are not intended to limit the same in any way.

[0089] Synthesis route of intermediate BB1

[0090] Step 1: Compound BB1-C tert-butyl 3-(4-chlorobenzyl)-2-oxopyrrolidine-1-carboxylate

[0091] To a solution of tert-butyl 2-oxopyrrolidine-1-carboxylate (4 g, 21.6 mmol) in tetrahydrofuran (60 mL) at -60°C under nitrogen was slowly added LiHMDS (21.6 ml, 21.6 mmol). After reacting at -60°C for 1 hour, a solution of 4-chlorobenzyl bromide (4.44 g, 21.6 mmol) in tetrahydrofuran (10 mL) was slowly added dropwise. The reaction mixture was allowed to react at -60°C for 2 hours, then warmed to room temperature and stirred for 18 hours. TLC monitored the reaction for completion. The reaction solution was quenched by adding saturated ammonium chloride solution (50 mL), extracted with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The resulting crude product was separated and purified by preparative thin-layer chromatography (eluent: 10-15% ethyl acetate in petroleum ether) to give BB1-C tert-butyl 3-(4-chlorobenzyl)-2-oxopyrrolidine-1-carboxylate (1.7 g) as a colorless oil.

[0092] 1 H NMR (400MHz, CDCl3) δ7.30–7.27(m,2H),7.15(d,J=8.4Hz,2H),3.69(ddd,J=11.2,8.7,2.7Hz,1H),3.59–3.51(m,1H),3. 23(dd,J=13.6,3.9Hz,1H),2.83–2.74(m,1H),2.68(t,J=6.8Hz,1H),2.06–1.99(m,1H),1.71–1.65(m,1H),1.55(s,9H).

[0093] Step 2: Compound BB1 3-(4-chlorobenzyl)pyrrolidin-2-one

[0094] Compound BB1-C tert-butyl 3-(4-chlorobenzyl)-2-oxopyrrolidine-1-carboxylate (1.7 g, 5.488 mmol) was added to a 4 M hydrochloric acid-dioxane solution (20 mL) at room temperature and stirred for 1 hour. The reaction was monitored for completion by TLC. The mixture was concentrated under reduced pressure to afford Compound BB1 3-(4-chlorobenzyl)pyrrolidin-2-one (1.15 g) as a white solid.

[0095] LC_MS:(ES + ):m / z 210.06[M+H] +

[0096] Synthesis route of intermediate BB2

[0097] Step 1: Compound BB2 1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0098] To a solution of BB2-A (1-(4-bromophenyl)pyrrolidin-2-one) (2.5 g, 10.41 mmol), BB2-B (1.92 g, 15.62 mmol), and sodium carbonate (2.76 g, 26.03 mmol) in acetonitrile (50 ml) and water (10 ml) was added Pd(PPh3)4 (1.2 g, 1.04 mmol). The reaction system was purged with nitrogen three times and heated to 90°C for 18 hours. TLC monitored the reaction for completion. The reaction mixture was cooled to room temperature and extracted with ethyl acetate and water. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: 0%-0.047% methanol in dichloromethane) to afford BB2 (1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (1.9 g) as a yellow solid.

[0099] LC_MS:(ES + ):m / z 239.05[M+H] + .

[0100] Synthesis route of intermediate BB3

[0101] Step 1: Compound BB3-C tert-butyl 3-(4-chlorobenzyl)-2-oxopiperidine-1-carboxylate

[0102] The synthesis of this step refers to the synthesis process of the first step of intermediate BB1.

[0103] LC_MS:(ES + ):m / z 324.13[M+H] + .

[0104] Step 2: Compound BB3 3-(4-chlorobenzyl)piperidin-2-one

[0105] The synthesis of this step refers to the synthesis process of the second step of intermediate BB1.

[0106] LC_MS:(ES + ):m / z 224.10[M+H] + .

[0107] Example 1: Synthesis route of compound 1

[0108] Step 1: Compound 1-C 1-(4-bromophenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one

[0109] Dissolve compound 1-A, 1-(4-bromophenyl)pyrrolidin-2-one (100 mg, 0.42 mmol), in anhydrous tetrahydrofuran (4 mL). Cool the system to -65°C under nitrogen. Then, add LDA (2 M in THF, 0.46 mL) dropwise at -65°C. Slowly raise the temperature to -30°C over half an hour, then lower it to -65°C. Add a solution of compound 1-B, 4-chlorobenzyl bromide (257 mg, 1.25 mmol) in THF (1 mL). Slowly return the reaction mixture to room temperature and continue the reaction for 1 hour. The reaction was monitored on a spot plate and completed. The reaction was quenched with a saturated ammonium chloride solution (10 mL), extracted with ethyl acetate (10 mL x 2), and the organic phases were combined, backwashed once with saturated brine (20 mL), dried and spin-dried. The crude product was separated and purified by silica gel column chromatography (eluted with a petroleum ether solution containing 6.7-16.7% ethyl acetate) to give compound 1-C as a white solid (1-(4-bromophenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one) (108 mg).

[0110] LC_MS:(ES + ):m / z 364.0[M+H] +

[0111] 1H NMR (400MHz, CDCl3): δ7.53-7.46(m,4H),7.28-7.26(m,2H),7.18-7.16(m,2H),3.76-3 .56(m,2H),3.25-3.21(m,1H),2.91-2.77(m,2H),2.22-2.15(m,1H),1.88-1.80(m,1H).

[0112] Step 2: Compound 1 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0113] Compound 1-C (1-(4-bromophenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one) (50 mg, 0.137 mmol), compound 1-D (pyridine-4-boronic acid) (21 mg, 0.164 mmol), sodium carbonate (29 mg, 0.27 mmol), and tetrakistriphenylphosphine palladium (16 mg, 0.014 mmol) were placed in a 10 mL reaction flask. Dioxane (1 mL) and H2O (0.5 mL) were then added. The atmosphere was purged with nitrogen three times and the reaction was allowed to proceed at 90°C overnight. The mixture was cooled to room temperature and the reaction was monitored by spot plate. Water was added to quench the mixture, and the mixture was extracted with ethyl acetate (10 mL x 2). The organic phases were combined, backwashed once with saturated brine (20 mL), dried and spin-dried. The crude product was separated and purified by Pre-TLC (eluted with dichloromethane solution containing 4% methanol) to give Compound 1 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (30.4 mg) as a white solid.

[0114] LC_MS:(ES + ):m / z 363.5[M+H] +

[0115] 1 H NMR (400MHz, CDCl3): δ8.66(d,J=5.2Hz,2H),7.79-7.76(m,2H),7.69-7.67(m,2H),7.55(d,J=6.4Hz,2H),7.29-7.27(m,2H),7.2 0-7.18(m,2H),3.80-3.65(m,2H),3.29-3.24(m,1H),2.98-2.91(m,1H),2.86-2.81(m,1H),2.27-2.19(m,1H),1.92-1.82(m,1H).

[0116] Example 2: Synthesis route of compound 2

[0117] Step 1: Compound 2-C 3-bromo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0118] To a mixture of 2-A 4-(pyridin-4-yl)aniline (50 mg, 0.294 mmol) and K3PO4 (32 mg, 0.147 mmol) in acetonitrile (2 mL) was added 2-B 2,4-dibromobutyryl chloride (78 mg, 0.294 mmol) under an ice bath. The ice bath was removed, and the reaction mixture was stirred at room temperature for 1 hour. Then, 50% aqueous NaOH (0.6 mL) was added. The reaction mixture was stirred overnight. The reaction mixture was filtered, the filter cake was washed with acetonitrile, and the filtrate was evaporated to dryness. The crude product was isolated and purified by silica gel column chromatography (eluting with 20-30% ethyl acetate in petroleum ether) to afford 2-C 3-bromo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg) as a yellow solid.

[0119] LC_MS:(ES + ):m / z 317.00[M+H] + .

[0120] Step 2: Compound 2 3-(4-chlorophenoxy)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0121] A solution of compound 2-C (3-bromo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (20 mg, 0.063 mmol), compound 2-D (4-chlorophenol) (9 mg, 0.063 mmol), and CsCO (41 mg, 0.126 mmol) in acetonitrile (2 mL) was heated to 60°C and stirred for 5 h. After completion of the reaction, the reaction solution was evaporated to dryness, and 20 mL of a mixed solvent of CHCl / MeOH (v / v = 20 / 1) was added for complete dissolution. The filtrate was then filtered and evaporated to dryness. The crude product was purified by Prep-TLC (eluent: CHCl / MeOH = 20 / 1) to afford compound 2 (3-(4-chlorophenoxy)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (11 mg) as a yellow solid.

[0122] LC_MS:(ES + ):m / z 365.40[M+H] + .

[0123] Example 3: Synthesis route of compound 3

[0124] Step 1: Compound 3-B N-(4-bromophenyl)hydroxylamine

[0125] To a solution of compound 3-A (1-bromo-4-nitrobenzene) (1.0 g, 4.95 mmol) in tetrahydrofuran (20 mL) was added rhodium on carbon (100 mg). The reaction mixture was cooled to 0°C, and hydrazine hydrate (309.77 mg, 4.95 mmol, 80% wt) was added portionwise, maintaining the addition temperature below 25°C. The reaction mixture was allowed to warm to room temperature and stirred for 2 hours. TLC monitored the reaction for completion. The reaction mixture was filtered through celite and concentrated under reduced pressure to yield the crude product, compound 3-B (N-(4-bromophenyl)hydroxylamine) (930.78 mg), which was used directly in the next step without further purification.

[0126] LC_MS:(ES + ):m / z 187.96[M+H] + .

[0127] Step 2: Compound 3-D 2-(4-bromophenyl)isoxazolin-3-one

[0128] To a solution of compound 3-B (N-(4-bromophenyl)hydroxylamine) (930.78 mg, 4.95 mmol) in N,N-dimethylformamide (10 mL) were added potassium carbonate (684.46 mg, 4.95 mmol) and compound 3-C (3-chloropropionyl chloride) (628.51 mg, 4.95 mmol). The reaction mixture was stirred at 25°C for 1 hour. LCMS confirmed the reaction was complete. The reaction mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL x 2). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was isolated and purified by silica gel column chromatography (eluent: 25% ethyl acetate in petroleum ether) to afford compound 3-D (2-(4-bromophenyl)isoxazolin-3-one) as a yellow solid (680 mg).

[0129] LC_MS:(ES + ):m / z 241.97[M+H] + .

[0130] 1 H NMR (400MHz, CDCl3) δ7.64–7.58(m,2H),7.53–7.47(m,2H),4.56(t,J=8.0Hz,2H),3.04(t,J=8.0Hz,2H).

[0131] Step 3: Compound 3-F 2-(4-(pyridin-4-yl)phenyl)isoxazolidin-3-one

[0132] To a solution of 3-D 2-(4-bromophenyl)isoxazolin-3-one (200 mg, 0.826 mmol), 3-E pyridine-4-boronic acid (152.44 mg, 1.24 mmol), and sodium carbonate (262.67 mg, 2.48 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was added Pd(dppf)Cl₂-CH₂Cl₂ (67.45 mg, 0.0826 mmol). The reaction system was purged with nitrogen three times and heated to 85°C for 18 hours. The reaction was complete, as monitored by TLC. The reaction solution was extracted with water (20 mL) and ethyl acetate (20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (eluent: 0%-0.047% methanol in dichloromethane) to give compound 3-F 2-(4-(pyridin-4-yl)phenyl)isoxazolidin-3-one (150 mg) as a yellow solid.

[0133] LC_MS:(ES + ):m / z 241.10[M+H] + .

[0134] Step 4: Compound 3 4-(4-chlorobenzyl)-2-(4-(pyridin-4-yl)phenyl)isoxazolidin-3-one

[0135] To a solution of compound 3-F 2-(4-(pyridin-4-yl)phenyl)isoxazolidin-3-one (50 mg, 0.208 mmol) in tetrahydrofuran (2.5 ml) was slowly added LDA (208 μl, 0.416 mmol) under nitrogen at -78°C. After 1 hour of reaction, compound 3-G 4-chlorobenzyl bromide (42.74 mg, 0.208 mmol) was added, and the reaction mixture was stirred at -78°C for 2 hours. TLC monitored the reaction for completion. The reaction mixture was quenched with saturated ammonium chloride solution (10 ml), extracted with ethyl acetate, and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was isolated and purified by preparative thin-layer chromatography (eluent: 0.047% methanol in dichloromethane) to afford compound 3-4-(4-chlorobenzyl)-2-(4-(pyridin-4-yl)phenyl)isoxazolidin-3-one (8 mg) as a yellow solid.

[0136] LC_MS:(ES + ):m / z 365.50[M+H] + .

[0137] Example 4: Synthesis route of compound 4

[0138] Step 1: Compound 4-C 1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0139] Under a nitrogen atmosphere, compound 4-A (4-(4-bromophenyl)pyridine) (120 mg, 0.5 mmol), compound 4-B (pyrrolidin-2-one) (60 mg, 0.75 mmol), cuprous iodide (10 mg, 0.05 mmol), potassium phosphate (212 mg, 1 mmol), and (1R,2R)-N,N'-dimethyl-1,2-cyclohexanediamine (14 mg, 0.1 mmol) were dispersed in 1,4-dioxane (1 mL). The reaction mixture was stirred at 100°C for 16 hours. LCMS analysis confirmed complete reaction of the starting material, with the target mass spectrum detected. TLC analysis revealed the formation of a major spot. The reaction solution was spin-dried to dryness, and dichloromethane (20 mL) was added. The reaction solution was washed sequentially with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to obtain compound 4-C 1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (80 mg) as a white solid.

[0140] LC-MS: (ES+): m / z 239.11 [M+H]+.

[0141] Step 2: Compound 4 3-(1-(4-chlorophenyl)ethyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0142] Compound 4-C 1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (26 mg, 0.1 mmol) was dissolved in 2 ml of tetrahydrofuran at -78°C, and lithium diisopropylamide (0.1 ml, 2N) was added dropwise. Half an hour later, a solution of compound 4-D 1-(1-bromoethyl)-4-chlorobenzene (26 mg, 0.12 mmol) in tetrahydrofuran was added dropwise. The reaction was stirred at low temperature for 1 hour. LCMS analysis indicated that the reaction of the starting material was complete, with the target mass spectrum detected, and TLC analysis indicated the formation of a major spot. Saturated ammonium chloride solution (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL×3). The reaction solution was washed with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL) in that order. The reaction solvent was removed by rotary evaporation, and the crude product was purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to obtain compound 4 (3-(1-(4-chlorophenyl)ethyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (10 mg) as a yellow solid.

[0143] LC-MS: (ES+): m / z 377.13 [M+H]+.

[0144] Example 5: Synthesis route of compound 5

[0145] Step 1: Compound 5-C 3-(4-chlorophenyl)-N-(4-(pyridin-4-yl)phenyl)propionamide

[0146] Under ice, a mixture of compound 5-A (3-(4-chlorophenyl)propionic acid) (184 mg, 1 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (210 mg, 1.1 mmol), and N,N-diisopropylethylamine (387 mg, 3 mmol) was dissolved in 5 mL of dichloromethane. After stirring for 15 minutes, compound 5-B (4-(4-pyridyl)aniline) (170 mg, 1 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. LCMS analysis confirmed the complete reaction of the starting material, with the target mass spectrum detected. TLC analysis revealed the formation of a major spot. Dichloromethane (20 mL) was added to the reaction solution, which was then washed sequentially with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL). The reaction solution was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by column separation and dried in vacuo to give a white solid compound 5-C3-(4-chlorophenyl)-N-(4-(pyridin-4-yl)phenyl)propionamide (250 mg).

[0147] LC-MS: (ES+): m / z 337.1 [M+H]+.

[0148] Step 2: Compound 5 2-(4-chlorobenzyl)-3-hydroxy-N-(4-pyridin-4-phenyl)propionamide

[0149] Compound 5-C, 3-(4-chlorophenyl)-N-(4-(pyridin-4-yl)phenyl)propanamide (34 mg, 0.1 mmol), was dissolved in 1 mL of tetrahydrofuran under an ice bath. Sodium hydride (60% wt, 9 mg, 0.22 mmol) was carefully added and stirred for 0.5 hour before addition of excess paraformaldehyde. The reaction mixture was allowed to warm to room temperature and continue stirring for 2 hours. LCMS analysis indicated product production. The reaction was quenched by the addition of saturated ammonium chloride solution (10 mL) and extracted with dichloromethane (10 mL x 3). The reaction solution was washed sequentially with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL). The solvent was removed by rotary evaporation. The crude product was separated and purified by preparative HPLC to yield Compound 5, 2-(4-chlorobenzyl)-3-hydroxy-N-(4-pyridin-4-phenyl)propanamide (1 mg), as a yellow solid.

[0150] LC-MS: (ES+): m / z 349.05 [M+H]+.

[0151] Example 6: Synthesis route of compound 6

[0152] Step 1: Compound 6-C 2-bromo-5-(pyridin-4-yl)pyridine

[0153] To a suspension of Compound 6-A (2-bromo-5-iodopyridine) (1.0 g, 3.53 mmol), Compound 6-B (pyridine-4-boronic acid) (434 mg, 3.53 mmol), and potassium carbonate (976 mg, 7.06 mmol) in 1,4-dioxane (15 ml) and water (3 mL) was added [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (25.9 mg, 0.035 mmol) under nitrogen at room temperature. The reaction mixture was purged with nitrogen three times and stirred at 85°C for 12 hours. TLC indicated the reaction was complete. The reaction mixture was cooled to room temperature, partitioned into water (20 mL), and extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by preparative TLC (eluted with 5% methanol in dichloromethane) to give compound 6-C 2-bromo-5-(pyridin-4-yl)pyridine (490 mg) as a white solid.

[0154] LC_MS:(ES + ):m / z 234.95,236.95[M+H] + .

[0155] 1 H NMR (400MHz, DMSO) δ8.88–8.85(m,1H),8.70(dd,J=4.5,1.7Hz,2H),8.20(dd,J=8.3,2.7Hz,1H),7.85–7.82(m,1H),7.82–7.79(m,2H).

[0156] Step 2: Compound 6 1-([3,4'-bipyridyl]-6-yl)-3-(4-chlorobenzyl)pyrrolin-2-one

[0157] To a suspension of compound 6-C (2-bromo-5-(pyridin-4-yl)pyridine) (170 mg, 0.723 mmol), compound 6-D (BB1) (3-(4-chlorobenzyl)pyrrolin-2-one) (150 mg, 0.723 mmol), (1R,2R)-N,N'-dimethyl-1,2-cyclohexanediamine (21 mg, 0.145 mmol), and potassium phosphate (307 mg, 1.446 mmol) in 1,4-dioxane (5 mL) was added cuprous iodide (14 mg, 0.073 mmol) at room temperature under a nitrogen atmosphere. The reaction mixture was purged with nitrogen three times and stirred at 100°C for 12 hours. TLC indicated the reaction was complete. The reaction mixture was cooled to room temperature, partitioned into water (20 mL), and extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated and purified by preparative TLC (eluted with a dichloromethane solution containing 9% methanol) to give compound 6 as a white solid, 1-([3,4'-bipyridyl]-6-yl)-3-(4-chlorobenzyl)pyrroline-2-one (209 mg).

[0158] LC_MS:(ES + ):m / z 364.10[M+H] + .

[0159] 1 H NMR (400MHz, CDCl3) δ8.87(d,J=2.5Hz,1H),8.75(m,2H),8.45(dd,J=8.7,2.7Hz,1H),8.01(d, J=4.4Hz,2H),7.89(d,J=8.8Hz,1H),7.30(t,J=5.4Hz,2H),7.21(d,J=8.4Hz,2H),3.83(dd,J=1 6.8,8.6Hz,1H),3.75(td,J=9.1,3.1Hz,1H),3.29(dd,J=13.7,4.2Hz,1H),2.99(qd,J=8.9,4. 2Hz,1H),2.87(dd,J=13.7,8.9Hz,1H),2.35–2.26(m,1H),1.95(ddd,J=18.0,12.8,8.8Hz,1H).

[0160] Example 7: Synthesis route of compound 7

[0161] Step 1: Compound 7-C 5-bromo-2,4'-bipyridine

[0162] The synthesis of this step uses compound 7-A 2,5-dibromopyridine as the raw material and refers to the synthesis process of the first step in Example 6.

[0163] LC_MS:(ES + ):m / z 234.95,236.95[M+H] + .

[0164] 1 H NMR (400MHz, CDCl3) δ8.82(d,J=2.3Hz,1H),8.76(d,J=6.1Hz,2H),7.98(dd,J=8.4,2.4Hz,1H),7.91(dd,J=4.7,1.5Hz,2H),7.74(d,J=8.4Hz,1H).

[0165] Step 2: Compound 7 1-([2,4'-bipyridine]-5-)-3-(4-chlorobenzyl)pyrrolidine-2-one

[0166] The synthesis of this step refers to the synthesis process of the second step in Example 6.

[0167] LC_MS:(ES + ):m / z 364.10[M+H] + .

[0168] 1 H NMR (400MHz, CDCl3) δ8.74(m,2H),8.67(d,J=2.1Hz,1H),8.61(d,J=8.8Hz,1H),8.00(dd,J=8.8 ,2.5Hz,1H),7.56(m,2H),7.32–7.29(m,2H),7.21(d,J=8.4Hz,2H),4.13(ddd,J=11.4,8.8,2.8H z,1H),3.92(ddd,J=11.2,8.9,7.4Hz,1H),3.31(dd,J=13.8,4.3Hz,1H),3.02(ddd,J=18.3,9.3, 4.4Hz, 1H), 2.83 (dd, J=13.8, 9.2Hz, 1H), 2.23 (tdd, J=10.3, 7.5, 2.8Hz, 1H), 1.92–1.83 (m, 1H).

[0169] Example 8: Synthesis route of compound 8

[0170] Step 1: Compound 8-C 3-bromo-6-(pyridin-4-yl)pyridazine

[0171] The synthesis of this step uses compound 8-A 3,6-dibromopyridazine as the raw material and refers to the synthesis process of the first step in Example 6.

[0172] LC_MS:(ES + ):m / z 235.90[M+H] + .

[0173] Step 2: Compound 8 3-(4-chlorobenzyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0174] The synthesis of this step refers to the synthesis process of the second step in Example 6.

[0175] LC_MS:(ES + ):m / z 241.10[M+H] + .

[0176] 1 H NMR(400MHz, CDCl3)δ8.87(d,J=9.4Hz,2H),7.96(d,J=9.4Hz,2H),7.32–7.29 (m,2H),7.23–7.18(m,4H),4.32(ddd,J=11.6,8.7,2.9Hz,1H),4.08(ddd,J=11 .5,9.0,7.4Hz,1H),3.30(dd,J=13.8,4.5Hz,1H),3.04(qd,J=9.1,4.5Hz,1H), 2.84(dd,J=13.9,9.1Hz,1H),2.36–2.24(m,1H),1.93(dq,J=12.8,9.1Hz,1H).

[0177] Example 9: Synthesis route of compound 9

[0178] Step 1: Compound 9-B(E)-1-(4-bromophenyl)-3-(dimethylamino)-2-propen-1-one

[0179] Compound 9-A (1 g, 5.02 mmol) was weighed and placed in a 25 mL reaction tube. 2 mL of DMF-DMA solvent was added and the mixture was heated to 80°C for 8 h. After the reaction was complete, the mixture was cooled to room temperature and poured into ice water. The solid precipitated and filtered. The filter cake was recrystallized from petroleum ether, filtered, and dried to obtain compound 9-B(E)-1-(4-bromophenyl)-3-(dimethylamino)-2-propen-1-one (1.0 g) as a yellow solid, which was directly used in the next step.

[0180] Step 2: Compound 9-C 4-(4-bromophenyl)pyrimidine

[0181] Compound 9-B(E)-1-(4-bromophenyl)-3-(dimethylamino)-2-propen-1-one (100 mg, 393.50 μmol) was placed in a 10 mL reaction tube. Formamidine acetate (49.2 mg, 472.20 μmol), sodium methoxide (150 μL, 5.4 M methanol solution), and ethanol (1.5 mL) were added sequentially at room temperature. The mixture was heated to 70°C and reacted overnight. After the reaction was completed, the mixture was cooled to room temperature, an appropriate amount of water was added, and the organic solvent was removed in vacuo. The mixture was extracted with ethyl acetate (10 mL x 3), dried over anhydrous sodium sulfate, and spin-dried. The yellow compound 9-C 4-(4-bromophenyl)pyrimidine (50 mg) was isolated and purified by preparative silica gel thin-layer chromatography (eluting with a 5% methanol in dichloromethane solution).

[0182] LC_MS:(ES + ):m / z 236.85[M+H] + .

[0183] Step 3: Compound 9 3-(4-chlorobenzyl)-1-(4-(pyrimidin-4-yl)phenyl)pyrrolidin-2-one

[0184] The synthesis of this step refers to the synthesis process of the second step in Example 6.

[0185] LC_MS:(ES + ):m / z 364.10[M+H] + .

[0186] 1 H NMR (400MHz, DMSO-d6) δ9.22(d,J=1.3Hz,1H),8.84(d,J=5.4Hz,1H),8.28-8.24(m,2H),8.09(dd,J=5.4,1.4Hz,1H),7.91-7.86(m,2H),7.3 9-7.30(m,4H),3.81-3.75(m,2H),3.16-3.11(m,1H),3.02-2.97(m,1H ),2.76(dd,J=13.6,9.3Hz,1H),2.12-2.06(m,1H),1.82-1.76(m,1H).

[0187] Example 10: Synthesis route of compound 10

[0188] Step 1: Compound 10-C 1-(4-bromo-3-methylphenyl)pyrrolidin-2-one

[0189] To a solution of compound 10-A (4-bromo-3-methylaniline) (500 mg, 2.72 mmol) and triethylamine (554 mg, 5.54 mmol) in dichloromethane (10 mL) was added dropwise 4-chlorobutyryl chloride (383 mg, 2.72 mmol) under ice-cooling conditions and stirred for 1 hour. After completion of the reaction, the reaction mixture was washed with water (20 mL x 2). The organic layer was concentrated and dissolved in tetrahydrofuran (10 mL). Sodium hydroxide (163 mg, 4.08 mmol) was added under ice-cooling conditions. The mixture was stirred for 2 hours. After the reaction was completed, the reaction solution was poured into a saturated ammonium chloride solution (20 mL), extracted with ethyl acetate (15 mL x 2), and the organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluted with petroleum ether solution containing 25% ethyl acetate) to give compound 10-C 1-(4-bromo-3-methylphenyl)pyrrolidin-2-one (330 mg) as a white solid.

[0190] LC_MS:(ES + ):m / z 254.1[M+H] + .

[0191] Step 2: Compound 10-E 1-(4-bromo-3-methylphenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one

[0192] To a solution of compound 10-C (1-(4-bromo-3-methylphenyl)pyrrolidin-2-one) (100 mg, 0.418 mmol) in tetrahydrofuran (2 mL) was added lithium diisopropylamide (0.418 ml, 0.836 mmol) at -78°C. The reaction mixture was stirred at -78°C for 1 hour. A solution of compound 10-D (4-chlorobenzyl bromide) (86 mg, 0.418 mmol) in tetrahydrofuran was then added dropwise, and the mixture was stirred at -78°C for another 2 hours. After the reaction, the reaction mixture was poured into saturated ammonium chloride (20 mL) and allowed to stand for separation. The organic phase was collected and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluted with 25% ethyl acetate in petroleum ether) to afford compound 10-E (1-(4-bromo-3-methylphenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one) (80 mg, 50%) as a pale yellow solid.

[0193] LC_MS:(ES + ):m / z 378.1[M+H] + .

[0194] Step 3: Compound 10 3-(4-chlorobenzyl)-1-(3-methyl-4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0195] A solution of compound 10-E 1-(4-bromo-3-methylphenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one (80 mg, 0.212 mmol), compound 10-F pyridine-4-boric acid (52 mg, 0.424 mmol), tetrakistriphenylphosphine palladium (2.5 mg, 2 μmol), and potassium carbonate (50 mg, 0.424 mmol) in N,N-dimethylformamide (2 mL) was heated to 85°C and stirred under nitrogen for 16 hours. After the reaction, the mixture was diluted with water (20 mL), extracted with ethyl acetate (10 mL x 2), and the organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluted with petroleum ether solution containing 25% ethyl acetate) to give Compound 10 3-(4-chlorobenzyl)-1-(3-methyl-4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg) as a white solid.

[0196] LC_MS:(ES + ):m / z 377.1[M+H] + .

[0197] Example 11: Synthesis route of compound 11

[0198] Step 1: Compound 11-C 1-(4-bromo-2-chlorophenyl)pyrrolidin-2-one

[0199] The synthesis of this step uses compound 10-A 4-bromo-2-chloroaniline as the raw material and refers to the synthesis process of the first step in Example 10.

[0200] LC_MS:(ES + ):m / z 274.02[M+H] + .

[0201] Step 2: Compound 11-E 1-(4-bromo-2-chlorophenyl)-3-(4-chlorobenzyl)pyrrolidin-2-one

[0202] The synthesis of this step refers to the synthesis process of the second step in Example 10.

[0203] LC_MS:(ES + ):m / z 398.10[M+H] + .

[0204] Step 3: Compound 11 3-(4-chlorobenzyl)-1-(2-chloro-4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0205] The synthesis of this step refers to the synthesis process of the third step of Example 10

[0206] LC_MS:(ES + ):m / z 397.10[M+H] + .

[0207] 1 H NMR (400MHz, DMSO-d6) δ8.71-8.66(m,2H),8.02(d,J=2.1Hz,1H),7.85(dd,J=8.2,2. 1Hz,1H),7.80-7.76(m,2H),7.53(d,J=8.2Hz,1H),7.44-7.31(m,4H),3.73-3.65(m, 1H),3.56(td,J=8.8,3.0Hz,1H),3.10(dd,J=13.7,4.4Hz,1H),2.92(qd,J=9.0,4.4H z,1H),2.76(dd,J=13.6,9.2Hz,1H),2.23-2.10(m,1H),1.86(dq,J=12.4,8.6Hz,1H).

[0208] Example 12: Synthesis route of compound 12

[0209] Step 1: Compound 12 3-(4-chlorophenyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidine-2-thione

[0210] To a solution of compound 1, 3-(4-chlorophenyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (50 mg, 137.80 umol) in toluene (2 mL), Lawesson's reagent (167.2 mg, 413.39 umol) was added. The reaction mixture was stirred at 100°C for 12 hours. TLC monitored the reaction for completion. The reaction mixture was concentrated, diluted with ethyl acetate, filtered, and the mother liquor was concentrated to obtain a residue which was purified by preparative thin-layer chromatography (eluent: 5% methanol in dichloromethane) to afford compound 12, 3-(4-chlorophenyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidine-2-thione (16.1 mg).

[0211] LC_MS:(ES + ):m / z 379.10[M+H] + .

[0212] Example 13: Synthesis route of compound 13

[0213] Step 1: Compound 13-C 1-(2-chloroethyl)-3-(4-(pyridin-4-yl)phenyl)urea

[0214] Compound 13-B chloroethyl isocyanate (308 mg, 2.94 mmol) was added dropwise to a solution of compound 13-A 4-(pyridin-4-yl)aniline (500 mg, 2.94 mmol) in dichloromethane (20 mL) under ice-cooling. The reaction mixture was stirred under ice-cooling for 1 hour. After completion of the reaction, the reaction solution was diluted with water (20 mL) and extracted with dichloromethane (20 mL x 2). The organic layers were combined and concentrated under reduced pressure. The residue was separated and purified by column chromatography (eluent: 50% ethyl acetate in petroleum ether) to obtain compound 13-C 1-(2-chloroethyl)-3-(4-(pyridin-4-yl)phenyl)urea (360 mg) as a white solid.

[0215] LC_MS:(ES + ):m / z 276.1[M+H] + .

[0216] Step 2: Compound 13-D 1-(4-(pyridin-4-yl)phenyl)imidazolin-2-one

[0217] To a solution of Compound 13-C (1-(2-chloroethyl)-3-(4-(pyridin-4-yl)phenyl)urea) (360 mg, 1.31 mmol) in tetrahydrofuran (5 mL) was added sodium hydride (104 mg, 2.62 mmol) under ice. The reaction mixture was stirred under ice for 3 hours. After completion of the reaction, the filter cake was collected and dried to afford Compound 13-D (1-(4-(pyridin-4-yl)phenyl)imidazolin-2-one) (300 mg) as a pale yellow solid.

[0218] LC_MS:(ES + ):m / z 240.1[M+H] + .

[0219] Step 3: Compound 13 1-(4-chlorobenzyl)-3-(4-(pyridin-4-yl)phenyl)imidazolin-2-one

[0220] Under ice bath conditions, sodium hydride (17.5 mg, 314 μmol) was added to a solution of compound 13-D 1-(4-(pyridin-4-yl)phenyl)imidazolin-2-one (50 mg, 209 μmol) in N,N-dimethylformamide (1 mL). After stirring for 30 minutes, compound 13-E 4-chlorobenzyl bromide (47 mg, 230 μmol) was added. The reaction mixture was stirred under ice bath for 3 hours. After the reaction was completed, the reaction solution was diluted with water (10 mL), filtered, and the filter cake was collected. The mixture was slurried with tetrahydrofuran (1 mL) for 2 hours, filtered, the filter cake was collected, and dried to obtain compound 13 1-(4-chlorobenzyl)-3-(4-(pyridin-4-yl)phenyl)imidazolin-2-one (30 mg) as a yellow solid.

[0221] LC_MS:(ES + ):m / z 364.1[M+H] + .

[0222] 1 H NMR(400MHz,Chloroform-d)δ8.66–8.60(m,2H),7.75–7.69(m,2H),7.69–7.62(m,2H),7.55–7.49( m,2H),7.37–7.30(m,2H),7.28(d,J=2.3Hz,2H),4.47(s,2H),3.91–3.82(m,2H),3.44–3.36(m,2H).

[0223] Example 14: Synthesis route of compound 14

[0224] Step 1: Compound 14 3-(4-chlorobenzyl)-1-(4-(3-methylpyridin-4-yl)phenyl)pyrrolidin-2-one

[0225] The synthesis of this step uses compound 14-A 3-methylpyridine-4-boronic acid as the raw material and refers to the synthesis process of the second step of Example 1.

[0226] LC_MS:(ES+):m / z 377.15[M+H]+.

[0227] 1H NMR(400MHz, CDCl3)δ8.52(s,1H),8.48(d,J=5.0Hz,1H),7.76–7.71(m,2H),7.40–7.35(m, 2H),7.33–7.29(m,2H),7.22(d,J=8.4Hz,2H),7.16(d,J=5.0Hz,1H),3.80(dd,J=16.8,8.5H z,1H),3.70(td,J=9.1,3.1Hz,1H),3.28(dd,J=13.7,4.1Hz,1H),2.96(ddd,J=17.6,8.9,4 .1Hz,1H),2.86(dd,J=13.7,8.9Hz,1H),2.32(s,3H),2.27–2.21(m,1H),1.93–1.85(m,1H).

[0228] Example 15: Synthesis route of compound 15

[0229] Step 1: Compound 15 3-(4-fluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0230] This step of the synthesis uses compound 15-A 4-fluorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step in Example 3.

[0231] LC_MS:(ES + ):m / z 347.45[M+H] + .

[0232] 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.1Hz,2H),7.79(d,J=8.8Hz,2H),7.69(d,J=8.8Hz,2H),7 .54(d,J=6.0Hz,2H),7.23(dd,J=8.5,5.5Hz,2H),7.02(t,J=8.7Hz,2H),3.79(dt,J=15.9,8 .0Hz,1H),3.68(td,J=9.1,3.3Hz,1H),3.28(dd,J=13.6,4.0Hz,1H),2.96(ddd,J=17.5,8. 8,4.1Hz,1H),2.86(dd,J=13.6,8.9Hz,1H),2.30–2.19(m,1H),1.89(dd,J=8.7,4.1Hz,1H).

[0233] Example 16: Synthesis route of compound 16

[0234] Step 1: Compound 16 3-(4-chloro-3-fluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0235] This step of the synthesis uses compound 16-A 3-fluoro-4-chlorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0236] LC_MS:(ES + ):m / z 381.05[M+H] + .

[0237] 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.7Hz,2H),7.79(d,J=8.7Hz,2H),7.69(d,J=8.7Hz,2H), 7.53(d,J=5.9Hz,2H),7.35(t,J=7.9Hz,1H),7.09(dd,J=9.9,1.5Hz,1H),7.01(d,J=8.2Hz ,1H),3.82(dd,J=16.7,8.9Hz,1H),3.74(td,J=9.1,2.7Hz,1H),3.29(dd,J=13.8,4.2Hz,1 H),3.01–2.92(m,1H),2.85(dd,J=13.8,9.0Hz,1H),2.31–2.23(m,1H),1.92–1.83(m,1H).

[0238] Example 17: Synthesis route of compound 17

[0239] Step 1: Compound 17 3-(4-methylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0240] This step of the synthesis uses compound 17-A 4-methylbenzyl bromide as the starting material and refers to the synthesis process of the fourth step in Example 3.

[0241] LC_MS:(ES + ):m / z 343.50[M+H] + .

[0242] 1H NMR (400MHz, CDCl3) δ8.67(d,J=6.0Hz,2H),7.79(d,J=8.8Hz,2H),7.69(d,J=8.8Hz, 2H),7.54(dd,J=4.7,1.5Hz,2H),7.20–7.10(m,4H),3.77(dt,J=15.8,7.9Hz,1H),3.6 9(td,J=9.1,3.4Hz,1H),3.30(dd,J=13.7,4.1Hz,1H),2.96(ddd,J=17.8,9.0,4.1Hz, 1H), 2.81 (dd, J=13.7, 9.3Hz, 1H), 2.35 (s, 3H), 2.28–2.19 (m, 1H), 1.91–1.86 (m, 1H).

[0243] Example 18: Synthesis route of compound 18

[0244] Step 1: Compound 18 3-(4-trifluoromethylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0245] This step of the synthesis uses compound 18-A 4-trifluoromethylbenzyl bromide as the starting material and refers to the synthesis process of the fourth step in Example 3.

[0246] LC_MS:(ES + ):m / z 397.50[M+H] + .

[0247] Example 19: Synthesis route of compound 19

[0248] Step 1: Compound 19 3-(4-cyanobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0249] The synthesis of this step uses compound 19-A 4-cyanobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0250] LC_MS:(ES + ):m / z 354.20[M+H] + .

[0251] 1H NMR (400MHz, CDCl3) δ8.68(d,J=5.1Hz,2H),7.79(d,J=8.6Hz,2H),7.70(d,J= 8.6Hz,2H),7.64(d,J=8.0Hz,2H),7.56(d,J=5.5Hz,2H),7.40(d,J=8.1Hz,2H) ,3.82(dd,J=16.7,8.8Hz,1H),3.73(td,J=9.1,2.6Hz,1H),3.38(dd,J=13.1,3 .7Hz,1H),3.06–2.90(m,2H),2.31–2.23(m,1H),1.91(dd,J=10.7,7.2Hz,1H).

[0252] Example 20: Synthesis route of compound 20

[0253] Step 1: Compound 20-C 2-bromo-5-(pyridin-4-yl)pyrazine

[0254] The synthesis of this step uses compound 20-A 2,5-dibromopyrazine as the raw material and refers to the synthesis process of the first step in Example 6.

[0255] LC_MS:(ES + ):m / z 235.95[M+H] + .

[0256] Step 2: Compound 20 3-(4-chlorobenzyl)-1-(5-(pyridin-4-yl)pyrazin-2-yl)pyrrolidin-2-one

[0257] The synthesis of this step refers to the synthesis process of the second step in Example 6.

[0258] LC_MS:(ES + ):m / z 365.10[M+H] + .

[0259] 1H NMR(400MHz, CDCl3)δ9.88(s,1H),8.83(s,1H),7.94(s,2H),7.42–7.12(m,6H),4.13–4.01(m,1H),3.95–3.81(m,1H),3.31(dd, J=13.9,4.3Hz,1H),3.04(qd,J=9.3,4.1Hz,1H),2.85(dd,J=13.9,9.1Hz,1H),2.27(dt,J=14.8,6.0Hz,1H),1.96–1.88(m,1H).

[0260] Example 21: Synthesis route of compound 21

[0261] Step 1: Compound 21-C 2-bromo-5-(pyridin-4-yl)benzonitrile

[0262] The synthesis of this step uses compound 21-A 2-bromo-5-iodobenzonitrile as the raw material and refers to the synthesis process of the first step in Example 6.

[0263] LC_MS:(ES + ):m / z 259.02[M+H] + .

[0264] Step 2: Compound 21 2-(3-(4-chlorobenzyl)-2-pyrrolidinone-1-yl)-5-(pyridin-4-yl)benzonitrile

[0265] The synthesis of this step refers to the synthesis process of the second step in Example 6.

[0266] LC_MS:(ES + ):m / z 388.20[M+H] + .

[0267] Example 22: Synthesis route of compound 22

[0268] Step 1: Compound 22-B 3-(4-chlorobenzyl)-3-phenylselenyl-1-(4-(4-pyridin-4-yl)phenyl)pyrrolidin-2-one

[0269] Under a nitrogen atmosphere, Compound 22-A (3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (180 mg, 0.5 mmol) was dispersed in 2 mL of tetrahydrofuran. Lithium diisopropylamide (0.4 mL, 2N) was added dropwise at -50°C. After 0.5 hour, a solution of phenylselenium bromide (140 mg, 0.6 mmol) in tetrahydrofuran was added dropwise. The reaction was stirred at low temperature for 1 hour. LCMS analysis revealed the target mass spectrometry, and TLC analysis revealed the formation of a major spot. The reaction was quenched by adding saturated ammonium chloride solution (20 mL), extracted with dichloromethane (10 mL x 3), and washed successively with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL). The reaction solvent was removed by rotary evaporation, and the crude product was purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to give compound 22-B 3-(4-chlorobenzyl)-3-phenylselenyl-1-(4-(4-pyridin-4-yl)phenyl)pyrrolidin-2-one (40 mg) as a white solid.

[0270] LC_MS:(ES + ):m / z 519.05[M+H] + .

[0271] Step 2: Compound 22 3-(4-chlorobenzyl)-1-(4-pyridin-4-yl)phenyl)-1,5-dihydro-2H-pyrrol-2-one

[0272] Compound 22-B, 3-(4-chlorobenzyl)-3-phenylselenyl-1-(4-(4-pyridin-4-yl)phenyl)pyrrolidin-2-one (40 mg, 0.1 mmol), was dissolved in 2 mL of dichloromethane at -78°C. Hydrogen peroxide (0.1 mL, 30% wt) was added dropwise. After 0.5 hour, one drop of pyridine (15 mg, 0.2 mmol) was added dropwise. The reaction was stirred at low temperature for another 0.5 hour, then allowed to warm to room temperature. LCMS analysis revealed the presence of the target mass. Saturated ammonium chloride solution (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL x 3). The reaction solution was washed with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL) in that order. The reaction solvent was removed by rotary evaporation, and the crude product was purified by reverse phase preparative column and lyophilized to give compound 22 (3-(4-chlorobenzyl)-1-(4-pyridin-4-yl)phenyl)-1,5-dihydro-2H-pyrrol-2-one) (5 mg) as a white solid.

[0273] LC_MS:(ES + ):m / z 361.50[M+H] + .

[0274] Example 23: Synthesis route of compound 23

[0275] Step 1: Compound 23-C 1-(2-bromopyrimidin-5-yl)pyrrolidin-2-one

[0276] The synthesis of this step uses compound 23-A 2-bromopyrimidin-5-amine as the starting material and refers to the synthesis process of the first step in Example 10.

[0277] LC_MS:(ES + ):m / z 241.90[M+H] + .

[0278] Step 2: Compound 23-E 1-(2-bromopyrimidin-5-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one

[0279] The synthesis of this step refers to the synthesis process of the second step in Example 10.

[0280] LC_MS:(ES + ):m / z 241.05[M+H] + .

[0281] Step 3: Compound 23 3-(4-chlorobenzyl)-1-(2-(pyridin-4-yl)pyrimidin-5-yl)pyrrolidin-2-one

[0282] The synthesis of this step refers to the synthesis process of the third step in Example 10.

[0283] LC_MS:(ES + ):m / z 365.10[M+H] + .

[0284] Example 24: Synthesis route of compound 24

[0285] Step 1: Compound 24 3-benzyl-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0286] The synthesis of this step uses compound 24-A benzyl bromide as the raw material and refers to the synthesis process of the fourth step of Example 3.

[0287] LC_MS:(ES + ):m / z 329.20[M+H] + .

[0288] Example 25: Synthesis route of compound 25

[0289] Step 1: Compound 25 3-((6-chloropyridin-3-yl)methyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0290] The synthesis of this step uses compound 25-A 5-(bromomethyl)-2-chloropyridine as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0291] LC_MS:(ES + ):m / z 364.11[M+H] + .

[0292] Example 26: Synthesis route of compound 26

[0293] Step 1: Compound 26 3-(3-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0294] This step of the synthesis uses compound 26-A 3-chlorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step in Example 3.

[0295] LC_MS:(ES + ):m / z 363.11[M+H] + .

[0296] Example 27: Synthesis route of compound 27

[0297] Step 1: Compound 27 3-(4-chlorophenethyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0298] This step of the synthesis uses compound 27-A 1-(2-bromoethyl)-4-chlorobenzene as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0299] LC_MS:(ES + ):m / z 377.13[M+H] + .

[0300] Example 28: Synthesis route of compound 28

[0301] Step 1: Compound 28 3-((5-chloropyridin-2-yl)methyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0302] The synthesis of this step uses compound 28-A 2-bromomethyl-5-chloropyridine as the raw material and refers to the synthesis process of the fourth step of Example 3.

[0303] LC_MS:(ES + ):m / z 364.11[M+H] + .

[0304] Example 29: Synthesis route of compound 29

[0305] Step 1: Compound 29 3-((5-chlorothien-2-yl)methyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0306] This step of the synthesis uses compound 29-A 2-(bromomethyl)-5-chlorothiophene as the starting material and refers to the synthesis process of the fourth step in Example 3.

[0307] LC_MS:(ES + ):m / z 369.10[M+H] + .

[0308] Example 30: Synthesis route of compound 30

[0309] Step 1: Compound 30 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0310] This step of the synthesis uses compound 30-A 3,4-difluorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0311] LC_MS:(ES + ):m / z 364.15[M+H] + .

[0312] Example 31: Synthesis route of compound 31

[0313] Step 1: Compound 31 3-(4-bromobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0314] The synthesis of this step uses compound 31-A 4-bromobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0315] LC_MS:(ES + ):m / z 407.08[M+H] + .

[0316] Example 32: Synthesis route of compound 32

[0317] Step 1: Compound 32 3-(4-nitrobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0318] The synthesis of this step uses compound 32-A 4-nitrobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0319] LC_MS:(ES + ):m / z 374.15[M+H] + .

[0320] Example 33: Synthesis route of compound 33

[0321] Step 1: Compound 33 3-(3,5-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0322] This step of the synthesis uses compound 33-A 3,5-difluorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0323] LC_MS:(ES + ):m / z 365.15[M+H] + .

[0324] Example 34: Synthesis route of compound 34

[0325] Step 1: Compound 34 3-(4-chlorobenzyl)-1-(4-(3-chloropyridin-4-yl)phenyl)pyrrolidin-2-one

[0326] The synthesis of this step uses compound 34-A 3-chloropyridine-4-boronic acid as the raw material and refers to the synthesis process of the second step of Example 1.

[0327] LC_MS:(ES+):m / z 397.12[M+H]+.

[0328] Example 35: Synthesis route of compound 35

[0329] Step 1: Compound 35 3-(4-chlorobenzyl)-1-(4-(3-fluoropyridin-4-yl)phenyl)pyrrolidin-2-one

[0330] The synthesis of this step uses compound 35-A 3-fluoropyridine-4-boronic acid as the starting material and refers to the synthesis process of the second step of Example 1.

[0331] LC_MS:(ES+):m / z 381.11[M+H]+.

[0332] Example 36: Synthesis route of compound 36

[0333] Step 1: Compound 36 3-(4-chlorobenzyl)-1-(4-(3-cyanopyridin-4-yl)phenyl)pyrrolidin-2-one

[0334] The synthesis of this step uses compound 36-A 3-cyanopyridine-4-boronic acid as the starting material and refers to the synthesis process of the second step of Example 1.

[0335] LC_MS:(ES+):m / z 388.11[M+H]+.

[0336] Example 37: Synthesis route of compound 37

[0337] Step 1: Compound 37 3-(4-chlorobenzyl)-1-(4-(nicotinamidopyridin-4-yl)phenyl)pyrrolidin-2-one

[0338] To a solution of 1,4-dioxane (1 mL) / water (1 mL) containing compound 36, 3-(4-chlorobenzyl)-1-(4-(3-cyanopyridin-4-yl)phenyl)pyrrolidin-2-one (50 mg, 128.91 umol), sodium carbonate (40.99 mg, 386.73 umol) was added. The reaction mixture was incubated at 80°C for 3 hours. LCMS confirmed the reaction was complete. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The residue was purified by preparative thin-layer chromatography (eluent: 10% methanol in dichloromethane) to afford compound 37, 3-(4-chlorobenzyl)-1-(4-(nicotinamidopyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg), as a yellow solid.

[0339] LC_MS:(ES+):m / z 406.12[M+H]+.

[0340] Example 38: Synthesis route of compound 38

[0341] Step 1: Compound 38 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)piperidin-2-one

[0342] To a solution of intermediate BB3 3-(4-chlorobenzyl)piperidin-2-one (47.78 mg, 213.59 umol) and compound 38-A 4-(4-bromophenyl)pyridine (50 mg, 213.59 umol) in dioxane (3 mL) were added Pd2(dba)3 (19.6 mg, 21.359 umol), Xant-Phos (12.4 mg, 21.359 umol), and cesium carbonate (208.77 mg, 640.76 umol). The reaction mixture was incubated at 100°C for 3 hours. LCMS monitoring confirmed the reaction was complete. The reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was separated and purified by preparative thin layer chromatography (eluent: 5% methanol in dichloromethane) to give yellow solid compound 38 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)piperidin-2-one (60 mg).

[0343] LC_MS:(ES+):m / z 377.12[M+H]+.

[0344] Example 39: Synthesis route of compound 39

[0345] Step 1: Compound 39-C 4-(4-bromo-3-methoxyphenyl)pyridine

[0346] This step of the synthesis uses compound 39-A 1-bromo-4-iodo-2-methoxybenzene as the starting material and refers to the synthesis process of the second step of Example 1.

[0347] LC_MS:(ES+):m / z 264.00[M+H]+.

[0348] Step 2: Compound 39 3-(4-chlorobenzyl)-1-(2-methoxy-4-(pyridin-4-yl)phenyl)piperidin-2-one

[0349] The synthesis of this step uses compound 39-C 4-(4-bromo-3-methoxyphenyl)pyridine as the raw material and refers to the synthesis process of the first step of Example 38.

[0350] LC_MS:(ES+):m / z 407.14[M+H]+.

[0351] Example 40: Synthesis route of compound 40

[0352] Step 1: Compound 40-B 2-(4-bromophenyl)-1,2-oxazol-3-one

[0353] The synthesis of this step uses compound 40-A 4-chlorobutyryl chloride as the starting material and refers to the synthesis process of the second step of Example 3.

[0354] LC_MS:(ES+):m / z 256.02[M+H]+.

[0355] Step 2: Compound 40-D 2-(4-(pyridin-4-yl)phenyl)-1,2-oxazol-3-one

[0356] The synthesis of this step uses compound 40-B 2-(4-bromophenyl)-1,2-oxazol-3-one as the raw material and refers to the synthesis process of the third step of Example 3.

[0357] LC_MS:(ES+):m / z 255.11[M+H]+.

[0358] Step 3: Compound 40 4-(4-chlorobenzyl)-2-(4-(pyridin-4-yl)phenyl)-1,2-oxazol-3-one

[0359] The synthesis of this step was carried out by using compound 40-D 2-(4-(pyridin-4-yl)phenyl)-1,2-oxazol-3-one as the starting material, and referring to the synthesis process of the fourth step of Example 3.

[0360] LC_MS:(ES+):m / z 379.11[M+H]+.

[0361] Example 41: Synthesis route of compound 41

[0362] Step 1: Compound 41 3-(2,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0363] This step of the synthesis uses compound 41-A 2,4-difluorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0364] LC_MS:(ES + ):m / z 365.15[M+H] + .

[0365] Example 42: Synthesis route of compound 42

[0366] Step 1: Compound 42 3-(4-chloro-2-fluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0367] This step of the synthesis uses compound 42-A 4-chloro-2-fluorobenzyl bromide as the starting material and refers to the synthesis process of the fourth step of Example 3.

[0368] LC_MS:(ES + ):m / z 365.15[M+H] + .

[0369] Example 43: Synthesis route of compound 43

[0370] Step 1: Compound 43-C(E)-1-(4-bromophenyl)-3-(4-chlorobenzylidene)pyrrolidine-2,5-dione

[0371] Compound 43-A N-(4-bromophenyl)maleimide (250 mg, 0.99 mmol) and compound 43-B (154 mg, 1.10 mmol) were dissolved in 10 mL of anhydrous ethanol. Triphenylphosphine (274 mg, 1.05 mmol) was added at room temperature and allowed to react overnight. After the reaction, the mixture was filtered, washed twice with ethanol, and the filter residue was vacuum dried to obtain compound 43-C(E)-1-(4-bromophenyl)-3-(4-chlorobenzylidene)pyrrolidine-2,5-dione (240 mg).

[0372] LC_MS:(ES + ):m / z 377.90[M+H]+.

[0373] Step 2: Compound 43-E(E)-3-(4-chlorobenzylidene)-1-(4-(pyridin-4-yl)phenyl)pyrrolidine-2,5-dione

[0374] The synthesis of this step uses compound 43-C(E)-1-(4-bromophenyl)-3-(4-chlorobenzylidene)pyrrolidine-2,5-dione as the raw material and refers to the synthesis process of the third step of Example 3.

[0375] LC_MS:(ES + ):m / z 375.15[M+H] + .

[0376] Step 3: Compound 43 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2,5-one

[0377] Compound 43-E(E)-3-(4-chlorobenzylidene)-1-(4-(pyridin-4-yl)phenyl)pyrrolidine-2,5-dione (12 mg) was weighed, and 2 mL of anhydrous ethanol, 2 mL of tetrahydrofuran, and 2 mg of Pd / C were added. The mixture was then replaced with hydrogen three times. The reaction was allowed to proceed overnight at room temperature. The mixture was separated by preparative thin-layer chromatography and purified by pre-HPLC to obtain compound 43 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidine-2,5-one (3 mg).

[0378] LC_MS:(ES + ):m / z 377.05[M+H] +

[0379] 1 H NMR(400MHz,Chloroform-d)δ8.71(d,J=5.1Hz,2H),7.77-7.72(m,2H),7.54-7.50(m,2H),7.36(dt,J=8.3,3.4Hz,4H),7.20(d,J=8.1Hz,2H),3. 37(dt,J=8.7,4.4Hz,1H), 3.26(dd,J=13.9,4.6Hz,1H), 3.11(dd,J=14.0,8.0Hz,1H), 2.96(dd,J=18.5,9.2Hz,1H), 2.66(dd,J=18.5,4.9Hz,1H).

[0380] Example 44: Synthesis route of compound 53

[0381] Step 1: Compound 53 3-(4-chlorobenzyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)piperidin-2-one

[0382] The synthesis of this step uses the intermediate BB3 3-(4-chlorobenzyl)piperidin-2-one and compound 8-C 3-bromo-6-(pyridin-4-yl)pyridazine as raw materials, and refers to the synthesis process of the first step in Example 38.

[0383] LC_MS:(ES + ):m / z 379.2[M+H] + .

[0384] 1H NMR(400MHz,DMSO-d6)δ8.81–8.75(m,2H),8.37(d,J=9.3Hz,1H),8.19–8.0 9(m,3H),7.39–7.34(m,2H),7.33–7.28(m,2H),4.14(ddd,J=12.9,8.3,4.9 Hz,1H),4.06–3.94(m,1H),3.25(dd,J=13.5,4.6Hz,1H),2.99–2.87(m,1H),2.78 (dd,J=13.6,9.0Hz,1H),2.06–1.76(m,3H),1.56(dtd,J=13.0,10.0,5.2Hz,1H).

[0385] Example 45: Synthesis route of compound 54

[0386] Step 1: Compound 54-C 3-(4-chlorobenzyl)azepan-2-one

[0387] Compound 54-A caprolactam (565 mg, 5 mmol) was dissolved in tetrahydrofuran (50 mL), cooled to 0°C under nitrogen, and n-butyllithium (6.4 mL, 10.25 mmol) was added. The mixture was reacted at 0°C for 2 hours. Then, compound 54-B 4-chlorobenzyl bromide (1130 mg, 5.5 mmol) dissolved in tetrahydrofuran (10 mL) was slowly added. The mixture was reacted at 0°C for 1 hour. The reaction was quenched by adding an excess of saturated aqueous ammonium chloride. The layers were separated, and the aqueous phase was extracted with ethyl acetate (30 mL). The organic phases were combined and washed with saturated aqueous sodium chloride (30 mL*2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound 54-C 3-(4-chlorobenzyl)azepan-2-one (142 mg) was isolated and purified by silica gel column chromatography (eluent: dichloromethane solution containing 7.5% methanol).

[0388] LC_MS:(ES + ):m / z 238.0[M+H] + .

[0389] Step 2: Compound 54 3-(4-chlorobenzyl)-1-(4-(pyridin-4-yl)phenyl)azepan-2-one

[0390] The synthesis of this step uses compound 54-C 3-(4-chlorobenzyl)azepan-2-one and compound 54-D 4-(4-bromophenyl)pyridine as raw materials, and refers to the synthesis process of the first step in Example 38.

[0391] LC_MS:(ES+ ):m / z 391.1[M+H] + .

[0392] 1 H NMR (400MHz, DMSO-d6) δ8.67–8.60(m,2H),7.84–7.77(m,2H),7.74–7.68(m,2H),7.35(d,J=5.7Hz,6H),4.12(dd,J=15.4,9.9Hz,1H),3.59(dd,J= 15.3,4.9Hz,1H),3.23(q,J=8.1Hz,2H),3.06(dd,J=14.0,6.6Hz,1H),2.00(q,J =7.3Hz,1H),1.85(d,J=15.0Hz,2H),1.74–1.54(m,2H),1.45(t,J=11.4Hz,1H).

[0393] Example 46: Synthesis route of compound 55

[0394] Step 1: Compound 55-B 1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0395] The synthesis of this step uses compound 8-C 3-bromo-6-(pyridin-4-yl)pyridazine and compound 55-A pyrrolidin-2-one as raw materials and refers to the synthesis process of the first step in Example 38.

[0396] LC_MS:(ES + ):m / z 241.10[M+H] + .

[0397] Step 2: Compound 55 3-(4-fluorobenzyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0398] This step of the synthesis uses compound 55-B 1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one and compound 55-C 1-(bromomethyl)-4-fluorobenzene as raw materials, referring to the synthesis process of the fourth step of Example 3.

[0399] LC_MS:(ES + ):m / z 349.12[M+H] + .

[0400] 1H NMR(400MHz,DMSO-d6)δ8.77(dd,J=4.5,1.7Hz,2H),8.73(d,J=9.5Hz,1H),8.43 (d,J=9.5Hz,1H),8.14–8.08(m,2H),7.38–7.29(m,2H),7.19–7.10(m,2H),4.23– 4.12(m,2H),3.96(ddd,J=10.9,8.9,7.4Hz,1H),3.13(ddt,J=17.9,9.1,4.6Hz, 2H), 2.78 (dd, J=13.3, 9.0Hz, 1H), 2.15 (q, J=8.1, 7.5Hz, 1H), 1.92–1.79 (m, 1H).

[0401] Example 47: Synthesis route of compound 58

[0402] Step 1: Compound 58 3-(4-chlorobenzyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidine-2-thione

[0403] The synthesis of this step uses compound 8 3-(4-chlorobenzyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one as the starting material and refers to the synthesis process of the first step in Example 12.

[0404] LC_MS:(ES + ):m / z 381.1[M+H] + .

[0405] 1 H NMR (400MHz, CDCl3) δ9.50(d,J=9.3Hz,1H),8.83(d,J=3.7Hz,2H),8.02(d,J=9.2Hz,3H),7.31(d,J=8.3Hz,2H),7.24(d,J=8.3Hz,2H),4.50(dd,J=8 .3,6.0Hz,2H),3.65(dd,J=13.8,4.1Hz,1H),3.34(ddd,J=18.0,8.5,4.2H z,1H),2.86–2.78(m,1H),2.31(dt,J=13.8,5.9Hz,1H),2.02–1.93(m,1H).

[0406] Example 48: Synthesis route of compound 59

[0407] Step 1: Compound 59-C 1-(4-chlorobenzyl)imidazolidin-2-one

[0408] To a DMF solution containing Compound 59-A imidazolidin-2-one (838 mg, 9.73 mmol) was added sodium hydride (116.8 mg, 4.87 mmol) under ice-cooling. The mixture was stirred for half an hour, followed by the dropwise addition of Compound 59-B 1-(bromomethyl)-4-chlorobenzene (1.0 g, 4.87 mmol). The mixture was stirred at room temperature for 12 hours. LCMS confirmed the reaction was complete. The reaction mixture was poured into saturated ammonium chloride solution (20 mL) and extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was isolated and purified by silica gel column chromatography (eluent: 50-70% ethyl acetate in petroleum ether) to afford Compound 59-C 1-(4-chlorobenzyl)imidazolidin-2-one (300 mg) as a yellow solid.

[0409] LC_MS:(ES + ):m / z 211.04[M+H] + .

[0410] Step 2: Compound 59 1-(4-chlorobenzyl)-3-(6-(pyridin-4-yl)pyridazin-3-yl)imidazolidin-2-one

[0411] The synthesis of this step uses compound 59-C 1-(4-chlorobenzyl)imidazolidin-2-one and compound 8-C 3-bromo-6-(pyridin-4-yl)pyridazine as raw materials, and refers to the synthesis process of the first step in Example 38.

[0412] LC_MS:(ES + ):m / z 366.10[M+H] + .

[0413] Example 49: Synthesis route of compound 74

[0414] Step 1: Compound 74-B 1-(6-bromopyridazin-3-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one

[0415] To a solution of compound 74-A (3,6-dibromopyridazine) (500 mg, 2.10 mmol) and intermediate BB1 ​​(3-(4-chlorobenzyl)pyrrolidin-2-one) (440.7 mg, 2.10 mmol) in dioxane (10 mL) were added Pd(dba) (96.2 mg, 105 umol), Xant-Phos (60.8 mg, 105 umol), and cesium carbonate (1.71 g, 5.25 mmol). The reaction mixture was allowed to react at 100°C under a nitrogen atmosphere for 2 hours. LCMS monitored the reaction for completion. The reaction mixture was cooled to room temperature, filtered, and concentrated under reduced pressure. The residue was purified by preparative thin-layer chromatography (eluent: 5% methanol in dichloromethane) to afford compound 74-B (1-(6-bromopyridazin-3-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one) (300 mg) as a yellow solid.

[0416] LC_MS:(ES + ):m / z 366.02[M+H] + .

[0417] Step 2: Compound 74 3-(4-chlorobenzyl)-1-(6-(3-methylpyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0418] This step of the synthesis uses compound 74-B 1-(6-bromopyridazin-3-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one and compound 74-C 3-methylpyridine-4-boronic acid as raw materials, referring to the synthesis process of the second step of Example 1.

[0419] LC_MS:(ES + ):m / z 379.10[M+H] + .

[0420] Example 50: Synthesis of Compound 75

[0421] Step 1: Compound 75 3-(4-chlorobenzyl)-1-(6-(3-fluoropyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0422] This step of the synthesis uses compound 74-B 1-(6-bromopyridazin-3-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one and compound 75-C 3-fluoropyridine-4-boronic acid as raw materials, and refers to the synthesis process of the second step of Example 1.

[0423] LC_MS:(ES + ):m / z 383.10[M+H] + .

[0424] Example 51: Synthesis route of compound 76

[0425] Step 1: Compound 76 3-(4-chlorobenzyl)-1-(6-(3-chloropyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0426] This step of the synthesis uses compound 74-B 1-(6-bromopyridazin-3-yl)-3-(4-chlorobenzyl)pyrrolidin-2-one and compound 76-A 3-chloropyridine-4-boronic acid as raw materials, and refers to the synthesis process of the second step of Example 1.

[0427] LC_MS:(ES + ):m / z 399.05[M+H] + .

[0428] Example 52: Synthesis of Compound 77

[0429] Step 1: Compound 77-B 3-(4-nitrobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0430] Intermediate BB2 1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (50 mg, 0.21 mmol) was dissolved in anhydrous tetrahydrofuran (4 mL) under nitrogen. The reaction mixture was stirred at -78°C for 5 minutes, and then 2M LDA (160 μL, 0.32 mmol) was slowly added dropwise. After the addition was complete, the reaction was continued for 30 minutes, followed by the dropwise addition of compound 77-A 1-(bromomethyl)-4-nitrobenzene (45 mg, 0.21 mmol). After the addition was complete, the reaction was continued at -78°C for 1 hour. After the reaction was completed, saturated aqueous ammonium chloride solution (5 mL) was added, and the reaction mixture was restored to room temperature, diluted with 20 mL of ethyl acetate, and the organic phase was retained. The organic phase was backwashed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and spin-dried. The mixture was separated and purified by preparative thin-layer chromatography to give a yellow solid compound 77-B 3-(4-nitrobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (26 mg).

[0431] Step 2: Compound 77-C 3-(4-aminobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0432] Compound 77-B 3-(4-nitrobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (26 mg) was dissolved in a mixed solvent of dichloromethane and methanol (4 mL, v / v = 1:1) and placed in an ice bath. Zinc powder (36 mg, 0.56 mmol) and ammonium chloride (37 mg, 0.69 mmol) were added sequentially. The temperature was slowly raised to room temperature and the reaction was allowed to proceed overnight. After the reaction was completed, solid impurities were removed by filtration and washed with dichloromethane and methanol (20 mL each). Purification by preparative thin-layer chromatography gave Compound 77-C 3-(4-aminobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg) as a yellow solid.

[0433] LC_MS:(ES + ):m / z 344.2[M+H] + .

[0434] Step 3: Compound 77 3-(4-(dimethylamino)benzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0435] Compound 77-C 3-(4-aminobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg) was dissolved in acetonitrile (2 mL) and placed in an ice bath. Paraformaldehyde (52 mg, 0.58 mmol), Et3SiH (34 mg, 0.29 mmol), and trifluoroacetic acid (33 mg, 0.29 mmol) were added sequentially, and the mixture was returned to room temperature and reacted overnight. After completion of the reaction, the organic solvent was removed in vacuo, methanol was added to dissolve the mixture, and saturated sodium bicarbonate solution was added to adjust the pH to 10-11. The solution was spin-dried and purified by two preparative thin-layer chromatography steps to obtain compound 77 3-(4-(dimethylamino)benzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (4 mg).

[0436] LC_MS:(ES + ):m / z 372.2[M+H] + .

[0437] 1H NMR(400MHz,Chloroform-d)δ8.68(d,J=5.3Hz,2H),7.83-7.75(m,2H),7.74-7.68(m,2H),7.59(d,J=5.5Hz,2H),7.15(d,J=8.5Hz,2H), 6.76(d,J=8.1Hz,2H),3.80-3.72(m,2H),3.24-3.20(m,1H),2.96-2.91(m,6H),2.80-2.75(m,1H),2.25-2.21(m,1H),1.94-1.90(m,1H).

[0438] Example 53: Synthesis route of compound 78

[0439] Step 1: Compound 78-B 3-((4-chlorophenyl)(hydroxy)methyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0440] Compound BB2 (1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (25 mg, 0.1 mmol) was dissolved in 10 mL of tetrahydrofuran at -78°C, and lithium diisopropylamide (1.5 eq, 0.075 ml, 2N) was added dropwise. After 0.5 hour, a solution of compound 78-A (4-chlorobenzaldehyde) (14 mg, 0.1 mmol) in tetrahydrofuran (10 mL) was added dropwise. The reaction was stirred at low temperature for 1 hour. LCMS analysis indicated that the reaction was complete, with the target mass detected. TLC analysis indicated the formation of a major spot. Saturated ammonium chloride solution (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 ml*3). The reaction solution was washed with water (15 mL) and saturated brine (20 mL) in that order. The reaction solvent was removed by rotary evaporation, and the crude product was purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to obtain yellow solid compound 78-B 3-((4-chlorophenyl)(hydroxy)methyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one (30 mg).

[0441] LC_MS:(ES + ):m / z 381.1[M+H] + .

[0442] Step 2: Compound 78 (E)-3-(4-chlorobenzylidene)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one

[0443] Compound 78-B, 3-((4-chlorophenyl)(hydroxy)methyl)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one (20 mg, 0.05 mmol) was dissolved in 1 mL of dichloromethane at 0°C. Triethylamine (25 mg, 0.25 mmol, 5 eq) was added dropwise. After 0.5 hour, a solution of methanesulfonyl chloride (11 mg, 0.1 mmol, 2 eq) in dichloromethane (0.5 mL) was added dropwise. The reaction was stirred at low temperature for 1 hour. LCMS analysis indicated that the reaction was complete and the target mass was detected. TLC analysis indicated the formation of a major spot. Water (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 ml*3). The reaction solution was washed with saturated brine (20 mL), and the reaction solvent was removed by rotary evaporation. The crude product was purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to give compound 78(E)-3-(4-chlorobenzylidene)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one (10 mg) as a white solid.

[0444] LC_MS:(ES + ):m / z 363.1[M+H] + .

[0445] Example 54: Synthesis route of compound 80

[0446] Step 1: Compound 80 3-((4-chlorophenyl)methyl-deuterium)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one-3-deuterium

[0447] Compound 78 (E)-3-(4-chlorobenzylidene)-1-(6-(pyridin-4-yl)pyridazin-3-yl)pyrrolidin-2-one (10 mg, 0.027 mmol) was dissolved in 1 mL of deuterated methanol at room temperature. A catalytic amount of platinum dioxide was added, and the mixture was stirred overnight at room temperature under a deuterium balloon. LCMS analysis indicated complete reaction, with the target mass spectrometry detected, and TLC analysis revealed a major spot. The solvent was dried by spin-drying, diluted with dichloromethane, and filtered. The solvent was then removed by rotary evaporation. The crude product was purified on preparative silica gel plates (developing solvent: DCM:MeOH = 20:1) and dried under vacuum to yield compound 80, 3-((4-chlorophenyl)methyl-deuterated)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one-3-deuterated (1 mg).

[0448] LC_MS:(ES + ):m / z 365.2[M+H] + .

[0449] Example 55: Chiral separation to prepare compound 81 and compound 82

[0450] Compound 30 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (50 mg) was dissolved in isopropanol and separated by an AD-H chiral column with a detection wavelength of 280 nm, a column temperature of 25°C, a pressure of 5 MPa, a mobile phase of n-hexane:isopropanol = 65:35, and a flow rate of 0.75 mL / min to obtain peak 1 (retention time: 19.52 min) and peak 2 (retention time: 25.78 min). The mixture was then concentrated under reduced pressure to give compound 81 (R)-3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one and compound 82 (R)-3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one.

[0451] Example 56: Synthesis route of compound 83

[0452] Step 1: Compound 83 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one-3-deuterium

[0453] Under nitrogen protection, a solution of compound 30 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (10.92 g, 30 mmol) in tetrahydrofuran (220 mL) was cooled to 0°C, and lithium bis(trimethylsilyl)amide (150 mL, 150 mmol) was added. The mixture was reacted at 0°C for 2 hours. The reaction solution was added to a -50°C solution of deuterated methanol-d4 (65 g, 1800 mmol), stirred for 10 minutes, and then gradually heated to 0°C for 3 hours. Saturated aqueous ammonium chloride solution (150 mL) was added to quench the mixture, and the pH was adjusted to about 8-9 with 1 M dilute hydrochloric acid. The liquid was separated, and the aqueous phase was extracted with ethyl acetate (100 mL*2). The organic phases were combined, washed with saturated brine (100 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product was slurried with ethanol (150 mL) for 1.5 hours, filtered, and the filter cake was washed with ethanol (30 mL*3). The filtered solid was dried to give compound 83 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one-3-deuterium (8.5 g, D%>99%) as a white solid.

[0454] LC_MS:(ES + ):m / z 365.8[M+H] + .

[0455] 1H NMR(400MHz,Chloroform-d)δ8.70–8.64(m,2H),7.81–7.76(m,2H),7.72–7.66(m,2H),7.55–7.49(m,2H),7.10(ddd,J=11.1,7.9,2.4Hz,2H),7.02 –6.95(m,1H),3.87–3.67(m,2H),3.27(d,J=14.1Hz,1H),2.84(d,J=14.1 Hz, 1H), 2.26 (ddd, J=10.9, 7.3, 3.0Hz, 1H), 1.89 (dt, J=12.8, 8.6Hz, 1H).

[0456] Example 57: Synthesis route of compound 84

[0457] Step 1: Compound 84 3-(3,4-difluorobenzyl)-3-hydroxy-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0458] Compound 30, 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (20 mg, 0.055 mmol), was dissolved in 10 mL of tetrahydrofuran at -78°C. Sodium bis(trimethylsilyl)amide (1.5 eq., 0.08 ml, 1 N) was added dropwise. After 0.5 hour, a solution of Davis reagent 3-phenyl-2-phenylsulfonyl-1,2-oxaziridine (21 mg, 0.08 mmol) in tetrahydrofuran (1 mL) was added dropwise. The reaction was stirred at low temperature for 2 hours. LCMS analysis indicated that the reaction of the starting material was complete, with the target mass spectrum detected. TLC analysis indicated the formation of a major spot. Saturated ammonium chloride solution (5 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL*3). The reaction solution was washed with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL) in that order. The reaction solvent was removed by rotary evaporation, and the crude product was purified by preparative silica gel plate (DCM:MeOH=20:1) and dried in vacuo to obtain compound 843-(3,4-difluorobenzyl)-3-hydroxy-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (5 mg) as a white solid.

[0459] LC_MS:(ES + ):m / z 381.2[M+H] + .

[0460] Example 58: Synthesis route of compound 85

[0461] Step 1: Compound 85 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)-1,5-dihydro-2H-pyrrol-2-one

[0462] Compound 84 (3-(3,4-difluorobenzyl)-3-hydroxy-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one) (20 mg, 0.05 mmol) was dissolved in 1 mL of dichloromethane at 0°C, and triethylamine (25 mg, 0.25 mmol, 5 eq) was added dropwise. After 0.5 hour, a dichloromethane solution (0.5 mL) of methylsulfonyl chloride (11 mg, 0.1 mmol, 2 eq) was added dropwise. The reaction was stirred at low temperature for 1 hour. LCMS analysis indicated that the reaction was complete, with the target mass detected. TLC analysis indicated the formation of a major spot. Saturated ammonium chloride solution (5 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL*3). The reaction solution was washed with water (15 mL), saturated ammonium chloride solution (20 mL), and saturated brine (20 mL) in that order. The reaction solvent was removed by rotary evaporation, and the crude product was purified by preparative silica gel plate (developing solvent: DCM:MeOH=20:1) and dried in vacuo to obtain compound 85 3-(3,4-difluorobenzyl)-1-(4-(pyridin-4-yl)phenyl)-1,5-dihydro-2H-pyrrol-2-one (2 mg) as a white solid.

[0463] LC_MS:(ES + ):m / z 363.0[M+H] + .

[0464] Example 59: Synthesis routes of Compound 86 and Compound 87

[0465] Step 1: Compound 86-C tert-butyl 3-(4-(methoxycarbonyl)benzyl)-2-oxopyrrolidine-1-carboxylate

[0466] To a solution of compound 86-A 1-(tert-butoxycarbonyl)-2-pyrrolidone (1.85 g, 10.0 mmol) in tetrahydrofuran (20 mL) was added dropwise 1M N,N-diisopropylamide lithium tetrahydrofuran solution (15.0 mL) under a nitrogen atmosphere at -78°C. The resulting mixture was stirred at -78°C for 1 hour, and then a solution of compound 86-B methyl 4-bromomethylbenzoate (2.29 g, 10.0 mmol) in tetrahydrofuran (30 mL) was slowly added dropwise. After reacting at -78°C for 1 hour, saturated ammonium chloride solution (20 mL) was added to quench the reaction, and the mixture was partitioned with dichloromethane (15 mL×3). The organic phase was collected and concentrated under reduced pressure. The crude product was separated and purified by silica gel column chromatography (25% ethyl acetate in petroleum ether as eluent) to give compound 86-C as a white solid. Tert-butyl 3-(4-(methoxycarbonyl)benzyl)-2-oxopyrrolidine-1-carboxylate (1 g).

[0467] LC_MS:(ES + ):m / z 234.1[M+H-100] + .

[0468] Step 2: Compound 86-D methyl 4-((2-oxopyrrolidin-3-yl)methyl)benzoate

[0469] To a solution of compound 86-C (tert-butyl 3-(4-(methoxycarbonyl)benzyl)-2-oxopyrrolidine-1-carboxylate) (1 g, 3.00 mmol) in dichloromethane (10 mL) was slowly added dropwise a 4M HCl solution in 1,4-dioxane (2 mL) under ice-cooling. The mixture was stirred on ice for 1.5 hours. After completion of the reaction as monitored by TLC, the mixture was concentrated under reduced pressure, and the pH was adjusted to neutral by adding saturated sodium bicarbonate solution. The mixture was partitioned with dichloromethane (5 mL x 3). The organic phases were combined, and the reaction mixture was concentrated under reduced pressure to yield crude compound 86-D (methyl 4-((2-oxopyrrolidin-3-yl)methyl)benzoate) (650 mg) as a white solid.

[0470] LC_MS:(ES+):m / z 234.1[M+H] + .

[0471] Step 3: Compound 86 methyl 4-((2-oxo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-3-yl)methyl)benzoate

[0472] A solution of compound 86-D (methyl 4-((2-oxopyrrolidin-3-yl)methyl)benzoate (300 mg, 1.28 mmol), compound 86-E (4-(4-bromophenyl)pyridine (300 mg, 1.28 mmol), tris(dibenzylideneacetone)dipalladium (108 mg, 0.0128 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (74 mg, 0.0128 mmol), and cesium carbonate (834 mg, 2.56 mmol) in 1,4-dioxane (5 mL) was reacted at 85°C under a nitrogen atmosphere for 12 hours. The reaction was monitored for completion by TLC. After the mixture was cooled to room temperature, it was filtered, and the filter residue was washed with dichloromethane (10 mL x 2). The combined filtrates were concentrated under reduced pressure, and the resulting crude product was isolated and purified by silica gel column chromatography (5% methanol in dichloromethane, eluent) to afford Compound 86 as a white solid, methyl 4-((2-oxo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-3-yl)methyl)benzoate (310 mg).

[0473] LC_MS:(ES + ):m / z 387.2[M+H] + .

[0474] 1 H NMR (400MHz, CDCl3) δ8.66(d,J=6.0Hz,2H),7.99(d,J=8.2Hz,2H),7.77(d,J=8.8Hz,2H),7. 67(d,J=8.8Hz,2H),7.52(dd,J=4.6,1.5Hz,2H),7.33(d,J=8.2Hz,2H),3.91(s,3H),3.78(dd ,J=16.6,8.9Hz,1H),3.68(td,J=9.1,3.1Hz,1H),3.37(dd,J=13.5,4.0Hz,1H),3.03–2.94( m,1H),2.90(dd,J=13.5,9.2Hz,1H),2.28–2.17(m,1H),1.88(ddd,J=17.9,12.7,8.7Hz,1H).

[0475] Step 4: Compound 87 4-((2-oxo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-3-yl)methyl)benzoic acid

[0476] To a solution of compound 86, methyl 4-((2-oxo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-3-yl)methyl)benzoate (50 mg, 0.129 mmol) in tetrahydrofuran / methanol / water (0.5 mL / 0.5 mL / 0.5 mL) was added sodium hydroxide (10.35 mg, 0.259 mmol) at room temperature. The reaction solution was heated to 50°C and stirred for 6 hours. LCMS monitored the reaction for completion. Dilute hydrochloric acid was added to the reaction solution to adjust the pH to approximately 3, producing a large amount of solid. The solid was collected by filtration to afford compound 87, 4-((2-oxo-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-3-yl)methyl)benzoic acid (45 mg), as a white solid.

[0477] LC_MS:(ES + ):m / z 373.1[M+H] + .

[0478] Example 60: Synthesis route of compound 88

[0479] Step 1: Compound 88-A 3-(4-(hydroxymethyl)benzyl)pyrrolidin-2-one

[0480] To a solution of Compound 86-D (methyl 4-((2-oxopyrrolidin-3-yl)methyl)benzoate (200 mg, 0.858 mmol) in tetrahydrofuran (8 mL) was slowly added lithium aluminum hydride (20 mg, 1.03 mmol) at 0°C. The reaction mixture was allowed to warm to room temperature and stirred for 3 hours. TLC monitored the reaction for completion. The reaction mixture was quenched with water (20 uL) and 10% sodium hydroxide solution (20 uL), filtered, and concentrated under reduced pressure. The resulting crude product was isolated and purified by preparative thin-layer chromatography (eluent: 10% methanol in dichloromethane) to afford Compound 88-A (3-(4-(hydroxymethyl)benzyl)pyrrolidin-2-one) (70 mg) as a beige solid.

[0481] LC_MS:(ES + ):m / z 206.1[M+H] + .

[0482] Step 2: Compound 88-B 4-(2-oxopyrrolidin-3-yl)methyl)benzaldehyde

[0483] To a solution of Compound 88-A, 3-(4-(hydroxymethyl)benzyl)pyrrolidin-2-one (70 mg, 0.341 mmol) in dichloromethane (5 ml), manganese dioxide (297 mg, 34.1 mmol) was slowly added. The reaction mixture was stirred at room temperature for 6 hours. TLC monitored the reaction for completion. The reaction mixture was filtered, the filter cake was washed with dichloromethane, and the filtrate was concentrated. The crude product was isolated and purified by column chromatography (eluent: 0%-5% methanol in dichloromethane) to afford Compound 88-B, 4-(2-oxopyrrolidin-3-yl)methyl)benzaldehyde (50 mg), as a beige solid.

[0484] LC_MS:(ES + ):m / z 204.1[M+H] + .

[0485] Step 3: Compound 88-D 3-(4-((4-methylpiperazin-1-yl)methyl)benzyl)pyrrolidin-2-one

[0486] A solution of Compound 88-B (4-(2-oxopyrrolidin-3-yl)methyl)benzaldehyde (50 mg, 0.246 mmol) and Compound 88-C (1-methylpiperazine) (27.1 mg, 0.27 mmol) in methanol (2.5 mL) was heated to 50°C and stirred for 1 hour. Sodium triacetylborohydride (104.27 mg, 0.492 mmol) was added, and the reaction mixture was stirred at 55°C for 24 hours. TLC confirmed the reaction completion. The reaction mixture was quenched with water (5 mL) and extracted with dichloromethane (5 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was isolated and purified by preparative thin-layer chromatography (eluent: 10% methanol in dichloromethane) to afford Compound 88-D (3-(4-((4-methylpiperazin-1-yl)methyl)benzyl)pyrrolidin-2-one) (33 mg) as a light yellow gum.

[0487] LC_MS:(ES + ):m / z 288.2[M+H] + .

[0488] Step 4: Compound 88 3-(4-((4-methylpiperazin-1-yl)methyl)benzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0489] Under nitrogen, Pd2(dba)3 (5.26 mg, 0.0058 mmol) was added to 1,4-dioxane (2 mL) containing compound 88-D 3-(4-((4-methylpiperazin-1-yl)methyl)benzyl)pyrrolidin-2-one (33 mg, 0.115 mmol), compound 88-E 4-(4-bromophenyl)pyridine (26.88 mg, 0.115 mmol), Xant-Phos (6.65 mg, 0.0115 mmol), and cesium carbonate (112.13 mg, 0.345 mmol). The reaction system was purged with nitrogen three times and heated to 100°C for 2 hours. The reaction was monitored for completion by TLC. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated. The crude product was separated and purified by preparative thin layer chromatography (eluent: 10% methanol in dichloromethane) to give Compound 88 3-(4-((4-methylpiperazin-1-yl)methyl)benzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (11.1 mg) as a white solid.

[0490] LC_MS:(ES + ):m / z 441.3[M+H] + .

[0491] 1 H NMR(400MHz, CDCl3)δ8.67(dd,J=4.6,1.6Hz,2H),7.81–7.76(m,2H),7.71–7.66(m,2 H),7.52(dd,J=4.6,1.6Hz,2H),7.24(t,J=9.0Hz,4H),3.79(dt,J=15.8,7.9Hz,1H), 3.73–3.66(m,1H),3.57(s,2H),3.34–3.29(m,1H),2.98(ddd,J=13.8,9.0,4.5Hz,1H ),2.89–2.81(m,1H),2.68(s,8H),2.48(s,3H),2.28–2.22(m,1H),1.95–1.88(m,1H).

[0492] Example 61: Synthesis route of compound 89

[0493] Step 1: Compound 89-C tert-butyl 3-(4-iodobenzyl)-2-oxopyrrolidine-1-carboxylate

[0494] This step of the synthesis uses Compound 89-A tert-butyl 2-oxopyrrolidine-1-carboxylate and Compound 89-B 1-(bromomethyl)-4-iodobenzene as raw materials, and refers to the synthesis process of the fourth step of Example 3.

[0495] LC_MS:(ES + ):m / z 402.0[M+H] + .

[0496] Step 2: Compound 89-E tert-butyl 2-oxo-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidine-1-carboxylate

[0497] To a solution of compound 89-C (tert-butyl 3-(4-iodobenzyl)-2-oxopyrrolidine-1-carboxylate) (200 mg, 0.5 mmol), bistriphenylphosphine palladium dichloride (10.5 mg, 0.015 mmol), and cuprous iodide (3.6 mg, 0.015 mmol) in tetrahydrofuran (4 mL) under nitrogen atmosphere was added compound 89-D (ethynyltrimethylsilane) (58.9 mg, 0.6 mmol) and triethylamine (151.8 mg, 1.5 mmol). The reaction mixture was stirred at room temperature for 4 hours. TLC monitored the reaction for completion. The reaction was quenched by adding water (10 mL), extracted with ethyl acetate (8 mL x 2), and the combined organic layers were dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The resulting crude product was separated and purified by silica gel column chromatography to give yellow solid Compound 89-E tert-butyl 2-oxo-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidine-1-carboxylate (176 mg).

[0498] LC_MS:(ES + ):m / z 372.2[M+H] + .

[0499] 1 H NMR (400MHz, CDCl3) δ7.41(d,J=7.9Hz,2H),7.15(d,J=8.0Hz,2H),3.66(dd,J=8.7,2.4Hz,1H),3.54(dd,J=9.4,7.5Hz,1H),3.26(dd,J=13 .6,3.8Hz,1H),2.83–2.75(m,1H),2.69(dd,J=13.6,9.5Hz,1H),2.00(m,1H),1.73–1.65(m,1H),1.58–1.54(m,9H),0.26(d,J=0.6Hz,9H).

[0500] Step 3: Compound 90-A (E) -3- (4- (2-chlorovinyl) benzyl) pyrrolidin-2-one and compound 89-F 3- (4- ( (trimethylsilyl) ethynyl) benzyl) pyrrolidin-2-one

[0501] To a solution of compound 89-E (tert-butyl 2-oxo-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidine-1-carboxylate) (176 mg, 0.474 mmol) in dichloromethane (2 mL) was added a hydrochloric acid / dioxane solution (1 mL). The reaction mixture was stirred at room temperature for 5 hours. TLC monitored the reaction for completion. The reaction mixture was concentrated, and the resulting crude product was separated and purified by preparative thin-layer chromatography (eluent: 8% methanol in dichloromethane) to afford compound 90-A(E)-3-(4-(2-chlorovinyl)benzyl)pyrrolidin-2-one (40 mg) and compound 89-F (3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidin-2-one (60 mg).

[0502] Compound 90-A_LC_MS: (ES + ):m / z 236.1[M+H] + .

[0503] Compound 89-F_LC_MS: (ES + ):m / z 272.1[M+H] + .

[0504] Step 4: Compound 89-H 1-(4-(pyridin-4-yl)phenyl)-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidin-2-one

[0505] The synthesis of this step uses compound 89-F 3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidin-2-one and compound 89-G 4-(4-bromophenyl)pyridine as raw materials, and refers to the synthesis process of the first step in Example 38.

[0506] LC_MS:(ES+):m / z 425.2[M+H]+.

[0507] Step 5: Compound 89 3-(4-ethynylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0508] Potassium carbonate (9.76 mg, 0.0706 mmol) was added to a solution of compound 89-H 1-(4-(pyridin-4-yl)phenyl)-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidin-2-one (15 mg, 0.353 mmol) in methanol (2 mL), and the reaction mixture was stirred at room temperature for 4 hours. TLC monitored the reaction for completion. The reaction mixture was concentrated, extracted with dichloromethane (8 mL x 2) and water (10 mL), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was isolated and purified by preparative thin-layer chromatography (eluent: 10% methanol in dichloromethane) to afford compound 89 3-(4-ethynylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (3.0 mg) as a white solid.

[0509] LC_MS:(ES + ):m / z 353.1[M+H] + .

[0510] 1 H NMR (400MHz, CDCl3) δ8.67(d,J=5.4Hz,2H),7.78(d,J=8.8Hz,2H),7.69(d,J=8.7Hz,2H),7 .53(d,J=6.0Hz,2H),7.46(d,J=8.1Hz,2H),7.24(d,J=8.1Hz,2H),3.78(dd,J=16.8,8.5Hz ,1H),3.69(td,J=9.1,3.0Hz,1H),3.33(dd,J=13.6,4.1Hz,1H),3.09(s,1H),2.98(ddd,J= 17.7,8.9,4.1Hz,1H),2.87(dd,J=13.6,9.1Hz,1H),2.29–2.19(m,1H),1.95–1.86(m,1H).

[0511] Example 62: Synthesis of Compound 90

[0512] Step 1: Compound 90 (E)-3-(4-(2-chlorovinyl)benzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0513] This step of the synthesis uses compound 90-A(E)-3-(4-(2-chlorovinyl)benzyl)pyrrolidin-2-one and compound 90-B 4-(4-bromophenyl)pyridine as raw materials, and refers to the synthesis process of the first step in Example 38.

[0514] LC_MS:(ES+):m / z 389.2[M+H]+.

[0515] Example 63: Synthesis route of compound 91

[0516] Step 1: Compound 91 3-(4-acetylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one

[0517] To a solution of compound 89-H 1-(4-(pyridin-4-yl)phenyl)-3-(4-((trimethylsilyl)ethynyl)benzyl)pyrrolidin-2-one (40 mg, 0.0942 mmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.5 mL), and the reaction mixture was stirred at room temperature for 4 hours. TLC monitored the reaction for completion. The reaction mixture was concentrated and extracted with saturated aqueous sodium bicarbonate (8 mL) and dichloromethane (5 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was isolated and purified by preparative thin-layer chromatography (eluent: 10% methanol in dichloromethane) to afford compound 91 3-(4-acetylbenzyl)-1-(4-(pyridin-4-yl)phenyl)pyrrolidin-2-one (10.3 mg) as a light yellow solid.

[0518] LC_MS:(ES + ):m / z 371.6[M+H] + .

[0519] 1 H NMR (400 MHz, CDCl3) δ8.67 (d, J = 5.8 Hz, 2H), 7.94 (d, J = 8.2 Hz, 2H), 7.79 (d, J = 8.7 Hz, 2H), 7.69 (d, J = 8.7 Hz, 2H), 7.53 (d, J = 6.1 Hz,2H),7.38(d,J=8.1Hz,2H),3.80(dd,J=16.9,8.5 Hz,1H),3.72(td,J=9.1,3.0 Hz,1H),3.39(dd,J=13.5,4.0 Hz,1H),3.02(ddd,J=17.6,8.9,4.0 Hz,1H),2.92(dd,J=13.5,9.1 Hz,1H),2.62(s,3H),2.30–2.20(m,1H),1.97–1.86(m,1H).

[0520] Synthetic routes of compounds 44-52

[0521] The synthetic routes of the following compounds are as follows, referring to the synthetic method of the fourth step of Example 3, using corresponding reagents for synthesis:

[0522] The synthetic routes of the following compounds are as follows, referring to the synthetic method of the second step of Example 46, using corresponding reagents for synthesis:

[0523] The following compounds can be synthesized by referring to the synthesis method of the first step of Example 56 using the corresponding reagents:

[0524] Biological activity test example:

[0525] Biological Activity Example 1: Preparation of SARM1 Enzyme

[0526] Preparation of test compounds:

[0527] The test compounds were prepared at a stock concentration of 200 uM or 10 mM (in DMSO) and further diluted to the desired compound concentration for in vitro SARM1 enzyme assays and inhibitor screening.

[0528] Expression and purification of SARM1 protein

[0529] (1) Plasmid construction

[0530] The gene sequence of SARM1 protein was amplified by PCR, and the PCR amplification product was constructed into the pcDNA3.1-HIS-C plasmid. The above construction was completed by Youbao Bio (Hunan, China).

[0531] (2) Plasmid transfection

[0532] HEK293 cells were cultured in a 10 cm dish using DMEM, and the next day the cells were transfected with 15 μg of pcDNA3.1-SARM1-HIS-C expression plasmid.

[0533] (3) Cell collection and protein extraction

[0534] 48 hours after transfection, cells were collected and the SARM1 protein expressed in the cells was obtained by diginotin cleavage for in vitro activity assays.

[0535] The cells were digested with trypsin-EDTA and centrifuged at 1000 rpm for 5 minutes. The cells were washed once with PBS and then resuspended in PBS containing 100 μM diginotin (0.6 mL PBS / 10 cm2 of cells) for 5 minutes. The cells were then treated with trypan blue and observed under a microscope, indicating that >90% of the cells had been lysed. The cells were then centrifuged at 5000 rpm for 10 minutes, and the supernatant containing SARM1 protein was collected.

[0536] Biological Activity Example 2: In vitro biochemical test for inhibition of SARM1 enzyme activity (IC 50 ):

[0537] First, 200 μM of the compound was added to a 50 mM Tris-HCl (pH 7.5) solution containing 0.05 μg / ml SARM1, and then half was added to an equal volume of 50 mM Tris-HCl (pH 7.5) solution containing 0.05 μg / ml SARM1 and mixed. The drug was diluted 6 times in this way, with final concentrations of 200, 100, 50, 25, 12.5, 6.25, 3.125 μM, or 200, 50, 12.5, 3.125, 0.78, 0.195, 0.049 μM. The control group was incubated at room temperature for 10 minutes without adding inhibitors.

[0538] Then, 50 μM NAD and 50 μM PC6 as substrates and 50 μM NMN as an activator were added to the SARM1 protein after incubation with the inhibitor and reacted at room temperature for 30 minutes. The concentrations of each component are the final concentrations in the reaction system.

[0539] During the reaction, PC6 fluorescence kinetics were monitored using a microplate reader, with excitation and emission wavelengths at 390 nm and 520 nm, respectively. The reaction rate was used to represent protein activity, and the 50% inhibitory concentration (50%) was calculated. Higher reaction rates indicate greater protein activity and lower compound inhibition efficiency.

[0540] The dosage curve of the compound for inhibition of SARM1 NAD enzyme activity was prepared using the above method.

[0541] The IC values ​​of these compounds in the assay are provided in Table 1 below. 50 Range:

[0542] IC for inhibition of SARM1 enzyme activity 50 Range: A<1μM; B:1-10μM; C:10-30μM

[0543] Table 1

[0544] Biological Activity Example 3: Axon Degeneration Index

[0545] Mouse DRG explant culture

[0546] Dorsal root ganglia were dissected from two-month-old C57BL / 6 male mice and placed in a 35 mm culture dish containing pre-chilled HBSS buffer. Excess tissue from each dorsal root ganglion was removed using a scalpel and dissected into 2-3 explants. These were seeded into wells of a 96-well plate coated with 100 ng / mL Matrigel (Corning). 100 μL of DRG culture medium (Neurobasal medium (Gibco) containing 2% B-27 (Gibco), 1% GlutaMax (Gibco), and 10 ng / mL NGF (MCE)) was added and the plates were cultured in a 37°C incubator for one week. The medium was changed on the fourth day, and experiments were performed on the eighth day.

[0547] Axonal Wallerian degeneration assay

[0548] The definition and measurement principle of the axonal degeneration index can be referred to the method reported in the prior art literature (Gerdts J, Sasaki Y, Vohra B, Marasa J, Milbrandt J. Image-based screening identifies novel roles for IkappaB kinase and glycogen synthase kinase 3 in axonal degeneration. J Biol Chem. 2011 Aug 12; 286(32): 28011-8). In the present invention, the specific determination method is as follows:

[0549] Take 0.1 μL of 10 mM test compound and dissolve it in 100 μL DRG culture medium to prepare a new culture medium containing 10 μM test compound. Pipette the culture medium in the 96-well plate and add the new culture medium containing the test compound, and incubate with the dorsal root ganglion explants in a 37-degree incubator for 2 hours for protection. After incubation, use a glass electrode to cut the axons in the area near the tissue block, and take bright field photos of the axons in the same area with a 20x microscope (Olympus) at 0 hours and 24 hours. The integrity of the axons was assessed using an internally developed script. Among them, the results are listed in Table 2 below according to the degree of protection (A): >50% and (B): 20%-50%.

[0550] The results showed that the following compounds all exhibited potent neuroprotective effects in the explant axon Wallerian degeneration assay at a concentration of 10 μM.

[0551] Table 2

Claims

1. Compound of formula (I): or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof, in, X1, X2, X3, X4, X5 are each independently selected from CH or N; Z represents -CH2-, -CH2(CH3)- or -O-; represents a single bond or a double bond; Y represents a carbon atom or a nitrogen atom; R1 is independently selected from H, halogen, C1-C3 alkyl, cyano, trifluoromethyl, amino, hydroxyl, methoxy-C(O)NH2, preferably H, F, Cl, methyl, cyano and -C(O)NH2; R2 is independently selected from H, halogen, C1-C3 alkyl, cyano, trifluoromethyl, amino, hydroxy, methoxy, C1-C3 alkyl-OC(O)-, C1-C3 alkyl-NHC(O)-, preferably H, cyano, Cl, methyl and methoxy; M stands for O or S; L is selected from -CH2-CH2-, -CH2-, -O-, -S-, -NH- and -CO-, wherein one -CH2- in the -CH2-CH2- can be optionally replaced by -O-, -S-, -CO- or -NH-; preferably, L is selected from -CH2-, -CH2-CH2-, -CH-O-, -O- and -CO-; E is selected from Wherein, X6, X7, X8 are each independently selected from CH or N; R3 is independently selected from H, halogen, methyl, cyano, trifluoromethyl, nitro, preferably H, F, Cl, Br, cyano, trifluoromethyl, nitro; R4 is independently selected from H, halogen, methyl, cyano, trifluoromethyl, and nitro, preferably H, F, Cl, Br, cyano, trifluoromethyl, and nitro.

2. The compound according to claim 1 or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof, wherein E is selected from wherein X6, X7, X8, R3 and R4 are as defined in claim 1.

3. The compound according to claim 1 or 2, or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof, wherein the compound of formula (I) has the following structure of formula (II): wherein X1, X2, X3, X4, X5, L, Y, R1, R3 and R4 are as defined in claim 1.

4. A compound or its racemate, enantiomer, diastereomer, deuterated form or a pharmaceutically acceptable salt thereof, wherein the compound is selected from the following compounds:

5. Use of a compound as claimed in any one of claims 1 to 4 or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing a neurodegenerative disease or a neurological disease or condition.

6. Use of the compound according to any one of claims 1 to 5 or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof in the preparation of a SARM1 enzyme activity inhibitor.

7. Use of the compound according to any one of claims 1 to 6 or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof in the preparation of a medicament for treating or preventing axonal degeneration-related diseases or disorders.

8. The method according to any one of claims 5 to 7, wherein the neurodegenerative disease or neurological disease or disorder or axonal degeneration-related disease or disorder is selected from Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, peripheral neuropathy.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or its racemate, enantiomer, diastereomer, deuterated substance or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

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