Compound for identifying alpha-synuclein aggregate and application thereof

Abstract

The invention relates to a compound for identifying alpha-synuclein aggregate and application thereof. The invention discloses a compound specifically combined with alpha-synuclein aggregate and a preparation method and application thereof. Specifically, the compound combined with the alpha-synuclein aggregate comprises a compound as shown in a formula A and a sub-general formula thereof, or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a stable isotope variant of the compound. The compound disclosed by the invention is a small molecule tracer agent, and can specifically recognize alpha-synuclein aggregate; the compound can be used for preparing medicines for treating or diagnosing neurodegenerative diseases (such as Parkinson's disease, dementia with Lewy bodies, multisystem atrophy, Alzheimer's disease, amyotrophic lateral sclerosis, progressive suprakaryotic paralysis, progressive amyotrophy and the like) related to alpha-synuclein aggregates and other misfolded proteins.

Description

[0001] Cross-reference

[0002] This application claims the priority of Chinese Patent Application No. 202410121172.8 with a filing date of January 29, 2024, Chinese Patent Application No. 202410121133.8 with a filing date of January 29, 2024, and Chinese Patent Application No. 202410209817.3 with a filing date of February 26, 2024. Technical field

[0003] The present invention relates to the field of medicine, and particularly to a compound that specifically recognizes α-synuclein aggregates and its uses. Background art

[0004] Neurodegenerative diseases (NDs) are diseases closely related to age, including Alzheimer's disease (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), multiple system atrophy (MSA), dementia with Lewy bodies (DLB), progressive muscular atrophy, progressive supranuclear palsy (PSP), etc. Their characteristics include progressive neuronal loss and degeneration. Currently, there are more than 57 million ND patients worldwide. With the increasing aging of the world's population, it is expected that by the middle of the 21st century, the number of ND patients will triple, which will lead to a huge medical and public health burden. However, at the present stage, there is no drug that can cure ND, and clinical medications can only relieve or slow down the symptoms of related diseases. ND has a long onset period, and there are pathological changes for more than a decade before clinical symptoms appear. During this process, most neurons of the patients have been lost. Given the non-renewability of neurons, diagnosing and treating early ND is an effective way to improve the curative effect of ND.

[0005] Abnormal protein aggregation is a common pathological change in ND and the most important early diagnostic biomarker for ND. Under physiological conditions, these proteins exist in a disordered monomer state, while under pathological conditions, these proteins will form an ordered β-sheet structure and gradually aggregate to form oligomers and fibrils. In AD, amyloid-β (Aβ) forms amyloid plaques (AP) outside cells, and these amyloid plaques are mainly deposited in the cortex, subcortical regions, and brainstem; at the same time, another pathological protein, Tau, can be fibrosed and form neurofibrillary tangles (NFT) deposited in the entorhinal cortex, hippocampus, and basal cortex. In PD, α-synuclein (α-syn) will form Lewy bodies and Lewy neurites, which appear in tissues and organs such as the substantia nigra, striatum, brainstem, and olfactory bulb.

[0006] Positron emission tomography (PET) and single photon emission computed tomography (SPECT) are non-invasive imaging methods that can detect the structure and function of the brain and thereby study the pathological progression and pathogenesis of ND. These techniques introduce radionuclides into small molecule tracers, which can target certain disease-related proteins in the body. The positrons or alpha photons emitted by the radionuclides can be detected and localized by the detectors of the imaging device, and the uptake of the tracer and its spatial distribution in the body can be calculated through a processor and image reconstruction software, from which changes in brain function and structure can be inferred. These techniques have relatively high spatial resolution, and the visualization and quantification resolution of PET can reach 3 mm.

[0007] Detecting α-synuclein aggregates in the brain by PET and SPECT techniques is of great significance for the early diagnosis of synucleinopathies. Synucleinopathies include PD, DLB, and MSA. In PD patients, abnormal neuronal inclusions called Lewy bodies (LB) and Lewy neurites (LN) appear in the brain, mainly in the substantia nigra and striatum. A large number of LB and LN are also present in the cerebral cortex of DLB patients. Different from PD and DLB, α-synuclein aggregates in the brains of MSA patients rarely exist in neurons but in the cytoplasm of oligodendrocytes and nerve cells, forming glial cytoplasmic inclusions (GCI). These pathological changes occur in the early stage of synucleinopathies and are closely related to the disease process. Therefore, imaging these misfolded α-synuclein by brain imaging techniques can achieve early diagnosis of the disease, which is of great significance for the treatment of the disease and the development of new drugs.

[0008] However, existing PET tracers have weak binding activity to proteins and poor in vivo imaging ability, unable to meet the needs of clinical imaging. Therefore, there is an urgent need to develop a compound that can specifically recognize α-synuclein aggregates, which is helpful for the early diagnosis and treatment of neurodegenerative diseases related to α-synuclein aggregates such as PD, DLB, and MSA in clinical practice. Summary of the Invention

[0010] To solve the above problems, the present invention provides a compound with strong binding ability to α-synuclein aggregates, which can be used to prepare preparations or preparation compositions for the treatment or diagnosis of neurodegenerative diseases related to α-synuclein aggregates and other misfolded proteins after binding to the target protein.

[0011] In a first aspect of the present invention, there is provided a compound that specifically recognizes α-synuclein aggregates, and the compound is a compound represented by formula A, or its stereoisomers, pharmaceutically acceptable salts, solvates, or stable isotope variants.

[0012]

[0013] Wherein each variable is as defined herein.

[0014] Specifically, the first aspect of the present invention provides a compound of the following sub-general formula, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof,

[0015]

[0016]

[0017] Wherein each variable is as generally or specifically defined herein.

[0018] The second aspect of the present invention provides a composition that specifically recognizes α-synuclein aggregates, and the composition includes

[0019] a. A compound as described in the first aspect of the present invention; and optionally

[0020] b. A pharmaceutically acceptable carrier.

[0021] The third aspect of the present invention provides the use of a compound as described in the first aspect of the present invention and a composition as described in the second aspect of the present invention in the preparation of a medicament for treating or diagnosing a neurodegenerative disease associated with α-synuclein aggregates and other misfolded protein aggregates.

[0022] The fourth aspect of the present invention provides a method for detecting α-synuclein aggregates in a subject, including the following steps:

[0023] (A) Administering a safe and effective amount of a compound as described in the first aspect of the present invention or a composition as described in the second aspect of the present invention to the subject; and

[0024] (B) Detecting the binding of the above compound or composition to α-synuclein aggregates in the subject.

[0025] In a preferred embodiment, the detection in step (B) is carried out by imaging techniques, preferably by the following imaging techniques: positron emission tomography, single photon emission computed tomography, near-infrared brain functional imaging, or a combination thereof.

[0026] In another preferred embodiment, the subject has or is suspected of having a neurodegenerative disease associated with α-synuclein aggregates or other misfolded protein aggregates.

[0027] In another preferred embodiment, the neurodegenerative disease is selected from the group consisting of: Parkinson's disease, multiple system atrophy, progressive muscular atrophy, dementia with Lewy bodies, Alzheimer's disease, progressive supranuclear palsy, or amyotrophic lateral sclerosis.

[0028] In a fifth aspect of the present invention, there is provided the use of a compound as described in the first aspect of the present invention and a composition as described in the second aspect of the present invention in the preparation of a kit for detecting the content of α-synuclein aggregates and other misfolded protein aggregates. Brief Description of the Drawings

[0029] Figure 1 The immunofluorescence staining diagram showing the co-localization of the compound of the present application with α-syn aggregates in primary neurons.

[0030] Figure 2 The immunofluorescence staining diagram showing the co-localization of the compound of the present application with α-syn aggregates in the brain of PFF mice (upper figure: Examples 1, 2, 3, 7, 11, 16, 26, 38 respectively refer to Examples II-1, II-2, II-3, II-7, II-11, II-16, II-26, II-38).

[0031] Figure 3 The immunofluorescence staining diagram showing the co-localization of the compound of the present application with α-syn aggregates in the brain of PD patients (upper figure: Examples 1, 2, 11, 26 respectively refer to Examples I-1, I-2, I-11, I-26; lower figure: Examples 1, 16, 19 respectively refer to Examples II-1, II-16, II-19).

[0032] Figure 4 The immunofluorescence staining diagram showing the co-localization of the compound of the present application with α-syn aggregates in the brain of PFF mice after intravenous injection of the compound of the present application into PFF mice (upper figure: Examples 1, 2 respectively refer to Examples I-1, I-2; lower figure: Examples 1, 16 respectively refer to Examples II-1, II-16).

[0033] Figure 5A and Figure 5B respectively show the autoradiography (ARG) maps of the compound of the present application in mouse and PD patient brain slices.

[0034] Figure 6A The PET imaging diagram of the compound 18F FD4 of the present application in PFF-injected rats is shown.

[0035] Figure 6B The PET imaging diagram of the compound 18F FD4 of the present application in PFF-injected marmosets is shown.

[0036] Figure 6C The PET imaging diagram of the compound FD17 of the present application in PFF-injected rats is shown.

[0037] Figure 6DShows the PET imaging of the compound FD17 of the present application in marmosets injected with PFF.

[0038] Figure 6E Shows the PET imaging of the compound FA35 of the present application in rats injected with PFF.

[0039] Figure 6F Shows the PET imaging of the compound FA23 of the present application in rats injected with PFF.

[0040] Figure 7 Shows the PET imaging of human subjects treated with the compound 18F FD4.

[0041] Figure 8 Shows the PET imaging of human subjects treated with C0505. Detailed Description of the Invention

[0043] Through long-term and in-depth research and extensive screening, the inventors have for the first time developed a compound that specifically recognizes α-synuclein aggregates, which includes compounds represented by Formula A and its respective sub-formulas, or their stereoisomers, pharmaceutically acceptable salts, solvates or stable isotope variants. The compounds of the present application can be used as detection reagents to detect the content of α-synuclein aggregates and other pathological aggregated proteins in in vitro samples, or for the preparation of drugs for the treatment or diagnosis of neurodegenerative diseases related to pathological aggregated proteins such as α-synuclein aggregates (such as Parkinson's disease, multiple system atrophy, dementia with Lewy bodies, Alzheimer's disease, amyotrophic lateral sclerosis, etc.), and have very good application prospects.

[0044] Terms

[0045] As used herein, the terms "comprising" or "including" can be open-ended, semi-closed and closed. In other words, the terms also include "consisting essentially of...", or "consisting of...".

[0046] As used herein, the term "alkyl" refers to a monovalent straight-chain or branched-chain saturated hydrocarbon group consisting only of carbon and hydrogen atoms. For example, "C 1-6 alkyl" represents an alkyl group having 1-6 (e.g., 1, 2, 3, 4, 5 or 6) carbon atoms or a range of carbon atoms composed of any different values therein, such as C 1-4 alkyl, C 1-3 alkyl, C 1-2 alkyl. Examples of alkyl groups include but are not limited to: methyl, ethyl, propyl, isopropyl, butyl, isobutyl or tert-butyl, etc.

[0047] As used herein, the term "alkylene" refers to a divalent group obtained by removing one hydrogen atom from the above-described alkyl group, such as methylene (-CH2-), ethylene (-CH2CH2-), etc.

[0048] As used herein, the term "alkenyl" refers to a straight-chain or branched-chain unsaturated hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one double bond. For example, "C 2-6 alkenyl" refers to an alkenyl having 2 to 6 (e.g., 2, 3, 4, 5, or 6) carbon atoms or a range of carbon atoms composed of any different values therein, such as C 2-4 alkenyl, C 2-3 alkenyl, C 2-5 alkenyl. Examples of alkenyl include, but are not limited to: vinyl, 1-propenyl, 2-propenyl, 1-butenyl, isobutenyl, and 1,3-butadienyl, etc.

[0049] As used herein, the term "alkynyl" refers to a straight-chain or branched-chain unsaturated hydrocarbon group composed of carbon atoms and hydrogen atoms and having at least one triple bond. For example, "C 2-6 alkynyl" refers to an alkynyl having 2 to 6 (e.g., 2, 3, 4, 5, or 6) carbon atoms or a range of carbon atoms composed of any different values therein, such as C 2-4 alkynyl, C 2-3 alkynyl, C 2-5 alkynyl. Examples of alkynyl include, but are not limited to: ethynyl, 1-propynyl, 2-propynyl, and 1-butynyl, etc.

[0050] As used herein, the term "cycloalkyl" refers to a monovalent saturated carbocyclic group composed of carbon and hydrogen atoms. For example, "C 3-8 cycloalkyl" refers to a cycloalkyl containing 3 to 8 (e.g., 3, 4, 5, 6, 7, or 8) carbon atoms or a range of carbon atoms composed of any different values therein, preferably C 3-6 cycloalkyl. The cycloalkyl can be monocyclic, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or similar groups, or can be in a bicyclic form, such as a bridged ring or spiro ring form, preferably in a monocyclic form.

[0051] As used herein, the term "haloalkyl" refers to a group obtained by substituting one or more hydrogens in the above-described alkyl group with the same or different halogens. Among them, "halo C 1-6 alkyl" or "C 1-6 haloalkyl" are used interchangeably, preferably halo C 1-4 alkyl. Examples of haloalkyl include, but are not limited to: -CH2Cl, -CH2CF3, -CH2CCl3, perfluoroalkyl (e.g., -CF3-, -CF2CF3), etc.

[0052] As used herein, the term "alkoxy" refers to a group of the formula -OR z or -R z '-OR z wherein R z is alkyl as defined herein, and R z ' is alkylene, preferably -C 1-4 alkoxy or -O-C 1-4 alkyl. Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, isopropoxy, tert-butoxy, -CH2O-CH3, -CH2CH2-O-CH3, -CH2-O-CH2CH3, and the like.

[0053] As used herein, the term "haloalkoxy" refers to a group obtained by substituting one or more hydrogens in the alkoxy group as described above with the same or different halogens, preferably -C 1-4 haloalkoxy or -O-C 1-4 haloalkyl. Examples of haloalkoxy groups include, but are not limited to: trifluoromethoxy, difluoromethoxy, 2,2,2-trifluoroethoxy, 2-fluoroethoxy, and the like.

[0054] As used herein, the term "halohydroxyalkoxy" refers to a group obtained by substituting one or more hydrogen atoms in the haloalkoxy group as described above with hydroxy groups, preferably -C 1-4 halohydroxyalkoxy or -O-C 1-4 halohydroxyalkyl, more preferably -C 1-4 haloalkoxy substituted with one hydroxy group. Examples of halohydroxyalkoxy groups include, but are not limited to

[0055] As used herein, the term "alkylamino" refers to a group of the formula -NR y R x wherein R y and R x are each independently H or alkyl as defined herein, and R y and R x are not both H at the same time, that is, alkylamino can be monoalkylamino or dialkylamino, preferably -NHC 1-4 alkyl or -N(C 1-4 alkyl)2. Examples of alkylamino groups include, but are not limited to: -NH-methyl, NH-ethyl, -N-dimethyl, -N-diethyl, and the like.

[0056] As used herein, the term "haloalkylamino" refers to a group obtained by substituting one or more hydrogens in the alkylamino group as described above with the same or different halogens, preferably -NHC 1-4 alkyl or -N(C 1-4 alkyl) 2，The alkyl groups are each independently substituted by one or more halogens, preferably by one or more F and / or Cl.

[0057] As used herein, the term "halo-hydroxyalkylamino" refers to a group obtained by substituting one or more hydrogen atoms in the haloalkylamino as described above with hydroxy groups, preferably -C substituted by 1 hydroxy group 1-4 haloalkylamino. Examples of halo-hydroxyalkylamino include, but are not limited to

[0058] As used herein, the term "halogen" refers to halogens and their isotopes, including but not limited to F, 18 F, Cl, 32 Cl, Br, I.

[0059] As used herein, the term "amino" refers to -NH2.

[0060] As used herein, the term "carboxyl" refers to -COOH.

[0061] As used herein, the term "ester group" refers to -COOR w wherein R w can be independently selected from the group consisting of: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic group; preferably -C(O)OC 1-6 alkyl. Examples of ester groups include, but are not limited to: -COOCH3, -COOCH2CH3, -COOCH2CH2CH3, -COOCH2CH(CH3)2, etc.

[0062] As used herein, the term "acyl group" refers to -COR w wherein R w can be independently selected from the group consisting of: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic group; preferably -C(O)C 1-6 alkyl. Examples of acyl groups include, but are not limited to: -COCH3, -COCH2CH3, -COCH2CH2CH3, -COCH2CH(CH3)2, etc.

[0063] As used herein, the term "amide group" refers to -CONR v R v ', wherein R v and R v' may be independently selected from the following group: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic group. R v and R v ' may be the same or different. Preferably, R v and R v ' are independently selected from hydrogen and C 1-6 alkyl, that is, the amide group is selected from -C(O)NH2, -C(O)NHC 1-6 alkyl, -C(O)N(C 1-6 alkyl)2. Examples of the amide group include, but are not limited to: -CONH2, -CONHCH3, -CON(CH3)2, etc.

[0064] As used herein, the term "sulfonamide group" refers to -SO2NR v R v ' or R v SO2NR v '- wherein R v and R v ' may be independently selected from the following group: hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocyclic group. R v and R v ' may be the same or different. Preferably, R v and R v ' are independently selected from hydrogen and C 1-6 alkyl, that is, the sulfonamide group is selected from -SO2NH2, -SO2NHC 1-6 alkyl, -SO2N(C 1-6 alkyl)2, -NHSO2-C 1-6 alkyl, -N(C 1-6 alkyl)-SO2-C 1-6 alkyl. Examples of the sulfonamide group include, but are not limited to: -SO2NH2, -SO2NHCH3, -SO2N(CH3)2, CH3SO2NH-, CH3SO2NCH3-, etc.

[0065] As used herein, the term "heterocyclic group" refers to a fully saturated or partially saturated monocyclic, bicyclic or polycyclic ring group containing one or more heteroatoms selected from N, S or O in the ring skeleton. For example, "3- to 12-membered heterocyclic group", "3- to 8-membered heterocyclic group", "4- to 8-membered heterocyclic group", "5- to 8-membered heterocyclic group", "6- to 8-membered heterocyclic group" refer to groups having 3 to 12 ring members, 3 to 8 ring members, 4 to 8 ring members, 5 to 8 ring members, and 6 to 8 ring members, respectively. Among them, the nitrogen atom or sulfur atom of the heterocyclic group can be oxidized, and the nitrogen atom can also be quaternized. The heterocyclic group can be attached to the residue of any heteroatom or carbon atom of the ring or ring system molecule. For the compounds of the present invention, a monocyclic 3- to 8-membered saturated heterocyclic group is preferred, such as a monocyclic 4- to 8-membered saturated heterocyclic group, a monocyclic 5- to 8-membered saturated heterocyclic group or a monocyclic 6- to 8-membered saturated heterocyclic group. Monocyclic heterocycles include but are not limited to: azetidinyl, pyrrolidinyl, oxetanyl, pyrazolinyl, imidazolinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, isothiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, hexahydroazepinyl, 4-piperidinonyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl, thiomorpholine sulfoxide, thiomorpholine sulfone, 1,3-dioxolanyl and tetrahydro-1,1-dioxothiophene, etc. Polycyclic heterocyclic groups include but are not limited to spiro, fused and bridged heterocyclic groups, wherein the spiro, fused and bridged heterocyclic groups involved are optionally connected to other groups by a single bond, or further fused to other cycloalkyl groups, heterocyclic groups, aryl groups and heteroaryl groups through any two or more atoms on the ring.

[0066] As used herein, the term "aryl" refers to an aromatic cyclic hydrocarbon group (including monocyclic, bicyclic or polycyclic groups), for example, "C 6-12 aryl" refers to an aromatic cyclic hydrocarbon group having 6 to 12 (6, 7, 8, 9, 10, 11 or 12) ring carbon atoms, and "C 6-10 aryl" refers to an aromatic cyclic hydrocarbon group having 6 to 10 (6, 7, 8, 9, 10) ring carbon atoms. Among them, when there are two or more aromatic rings (such as bicyclic, etc.), the aromatic rings of the aryl group can be connected by a single bond (such as biphenyl) or fused (such as naphthalene, anthracene, etc.). Examples of aryl groups (especially monocyclic and bicyclic groups) include but are not limited to: phenyl, biphenyl or naphthyl. The aryl group can be fused to the heterocyclic group by a single bond or any two adjacent ring carbon atoms. For example: benzotetrahydrofuranyl, benzotetrahydropyranyl, benzodioxanyl, etc.

[0067] As used herein, the term "heteroaryl" refers to an aromatic cyclic group (including monocyclic, bicyclic or polycyclic groups) whose ring skeleton contains 1, 2, 3 or 4 heteroatoms selected from N, S or O. For example, "5-12 membered heteroaryl" refers to a monocyclic, bicyclic or tricyclic group having 5 to 12 (5, 6, 7, 8, 9, 10, 11 or 12) ring atoms, and "5-10 membered heteroaryl", "5-8 membered heteroaryl", "6-10 membered heteroaryl" refer to monocyclic or bicyclic groups having 5 to 10 (5, 6, 7, 8, 9, 10, 11 or 12), 5-8 or 6-10 ring atoms respectively. Examples of heteroaryl include but are not limited to: imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, pyrazinyl, thienyl, furyl, pyranyl, pyridyl, pyrrolyl, pyrazolyl, pyrimidinyl, quinolinyl, isoquinolinyl, benzofuryl, benzothienyl, benzothiopyranyl, benzimidazolyl, benzoxazolyl, benzoxadiazolyl, benzothiazolyl, benzothiadiazolyl, benzopyranyl, indolyl, isoindolyl, triazolyl, triazinyl, quinoxalinyl, purinyl, quinazolinyl, quinazylinyl, naphthyridinyl, pteridinyl, carbazolyl, azanyl, diazanyl, acridinyl, etc. yl yl, etc.

[0068] As used herein, the term "substituted" means that one or more hydrogen atoms on a particular group are replaced by a particular substituent. The particular substituent is the substituent described correspondingly in the foregoing, or the substituent appearing in each embodiment. Unless otherwise specified, a substituted group may have a substituent selected from a particular group at any substitutable site of the group, and the substituents may be the same or different at each position. Those skilled in the art should understand that the combinations of substituents contemplated by the present invention are those that are stable or chemically achievable.

[0069] Unless otherwise specified as substituted or unsubstituted, the groups described in the present invention can be substituted by substituents selected from the following group: D, halogen, cyano, nitro, hydroxy, amino, -C 1-6 alkyl, -C 2-6 alkenyl, -C2-6 alkynyl, -C 1-6 alkoxy, 3-12 membered heterocyclic group, C3-C 12 cycloalkyl, 5-10 membered heteroaryl and -C 6-10 aryl.

[0070] As used herein, "optionally" means that the subsequently described event or condition may but does not have to occur, and this description includes the case where the event or condition occurs and the case where the event or condition does not occur.

[0071] As used herein, the term "plurality" refers to a positive integer of 2, 3, 4, 5 or greater than 5.

[0072] As used herein, the terms "α-synuclein aggregate", "α-syn aggregate", and "α-syn fibril" are synonymous.

[0073] As used herein, the terms "pathological aggregate protein" and "misfolded protein" are synonymous.

[0074] As used herein, "the compounds of the present invention" refers to the compounds represented by formula (A) and each of its sub-formulas, and also includes their stereoisomers, their optical isomers, their pharmaceutically acceptable salts, their crystal forms, their isotope derivatives, their prodrugs, their metabolites, their solvates, or their hydrates.

[0075] As used herein, the term "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0076] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic acids or organic acids that can retain the biological effectiveness of the free base without other side effects. Inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include, but are not limited to, formate, acetate, 2,2-dichloroacetate, trifluoroacetate, propionate, caproate, caprylate, caprate, undecylenate, glycolate, gluconate, lactate, sebacate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylate, naphthalenedisulfonate, etc. These salts can be prepared by methods known in the art.

[0077] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that can maintain the biological effectiveness of the free acid without other side effects. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, etc. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, the following salts: primary amines, secondary amines, and tertiary amines, substituted amines, including naturally substituted amines, cyclic amines, and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, triethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Preferred organic bases include isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine. These salts can be prepared by methods known in the art.

[0078] As used herein, the term "solvate" refers to a complex formed by the coordination of a compound represented by formula (A) and its respective sub-formulas with solvent molecules in a specific ratio.

[0079] As used herein, the term "safe and effective amount" means an amount of a compound sufficient to treat or diagnose a disease without causing serious side effects.

[0080] As used herein, the term "pharmaceutically acceptable carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatible" herein means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds.

[0081] The compounds of the present invention

[0082] Unless otherwise specified, the structural formulas described in the present invention are intended to include all stereoisomers (such as cis-trans isomers, enantiomers, diastereomers, and conformational isomers): the R and S configurations containing asymmetric centers, the (Z) and (E) isomers of double bonds, the cis-trans isomers of cycloalkanes, etc. Therefore, single stereochemical isomers of the compounds of the present invention or mixtures of their enantiomers, diastereomers, cis-trans isomers, or conformational isomers are all within the scope of the present invention.

[0083] The compounds of the present invention may contain cis-trans isomers, one or more chiral carbon atoms, and thus can produce stereoisomeric forms such as cis-trans isomers, chiral isomers, enantiomers, diastereomers, and other combinations.

[0084] Cis-trans isomerism refers to a diastereoisomerism phenomenon that occurs in a compound molecule due to restricted factors for free rotation, resulting in different arrangements of various groups in space. Such restricted factors are generally caused by functional groups that cannot rotate freely in the structure of organic compounds, such as C=C double bonds, C=N double bonds, C=S double bonds, N=N double bonds, heterocycles or cycloalkanes. Organic molecules containing such isomers, such as alkenes, azo compounds, and alicyclic hydrocarbons, are regarded as cis-trans isomers. The cis form means that the same ligands are in adjacent positions, generally represented by "cis-" or "cis"; the trans form means that the same ligands are in diagonal positions, generally represented by "trans-" or "trans".

[0085] Each chiral carbon atom can be defined as (R)- or (S)- based on stereochemistry. The present invention aims to include all possible isomers, their racemates, any proportion of enantiomeric mixtures, or optically pure forms. It should be noted that in the case where the compounds of the present invention contain only one chiral center, the mention of the racemate in this application is equivalent to simultaneously mentioning the two enantiomers present therein, and those skilled in the art can prepare the individual enantiomers based on the disclosure in the context of the specification and conventional means in the art.

[0086] For the preparation of the compounds of the present invention, racemates, cis-trans isomers, chiral isomers, diastereoisomers or enantiomers can be selected as raw materials or intermediates. Optically active isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0087] Conventional techniques for preparing / separating individual optical isomers (i.e., cis-trans isomers and chiral / enantiomeric isomers) include chiral synthesis from suitable cis-trans precursors or optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high performance liquid chromatography.

[0088] If the synthesis of a specific stereoisomer of a compound of the present invention is to be designed, it can be prepared by asymmetric synthesis, or derivatized with a chiral auxiliary, the resulting stereomix is separated, and then the chiral auxiliary is removed to obtain pure cis-trans monomers, chiral monomers or mixed stereomonomers. If the molecule contains cis-trans isomeric centers, pure cis or trans products can be obtained by column chromatography purification (normal phase silica gel column or reverse phase high performance liquid preparation); alternatively, if the molecule contains a basic functional group, such as an amino acid, or an acidic functional group, such as a carboxyl group, it can be formed into a diastereoisomeric salt with a suitable optically active acid or base, and then separated by conventional means such as fractional crystallization or chromatography, and then pure enantiomers are obtained.

[0089] The present invention also includes isotopically labeled compounds (i.e., isotopic derivatives), which are equivalent to the original compounds disclosed herein. However, in practice, substitution of one or more atoms with atoms having a different atomic weight or mass number usually occurs. Examples of isotopes in the isotopic derivatives of the compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, such as 2 H, 3 H, 13 C, 11 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl, respectively. Isotopic derivatives of the compounds of the present invention are all within the scope of protection of the present invention.

[0090] In some embodiments, the compounds of the present invention are isotopically 18 F-labeled compounds, as shown in some representative examples of the present invention. It should be noted that although some of the compounds in the embodiments of the present application have not been isotopically labeled, those skilled in the art can prepare isotopically labeled forms of the compounds according to the content disclosed in the present application, such as 18F-labeled forms, and these compounds are also covered by the present application.

[0091] In some embodiments, the compounds of the present invention can be 3H-labeled compounds and 14 C-labeled compounds, which are useful in tissue distribution experiments of drugs and substrates. Tritium (i.e., 3 H) and carbon- 14 (i.e., 14 C)-labeled compounds are relatively easy to prepare and detect and are preferred among isotopes.

[0092] In some embodiments, the compounds of the present invention can have heavier isotope substitutions, such as deuterium, i.e., 2H, which has advantages in certain therapies due to its good metabolic stability, such as increasing the half-life or reducing the dosage in vivo. Therefore, it can be preferred in certain cases.

[0093] Isotopically labeled compounds can be prepared by general methods, by replacing non-isotopic reagents with readily available isotopic labeling reagents, using the procedures disclosed in the examples.

[0094] Metabolites of the compounds represented by formula (A) and its respective sub-formulas, and their pharmaceutically acceptable salts, as well as prodrugs that can be converted in vivo into the compounds represented by formula (A) and its respective sub-formulas and their pharmaceutically acceptable salts, are also included within the scope of protection of the present invention.

[0095] Specifically, the present invention provides the following compound embodiments:

[0096] Embodiment 1: A compound of formula A, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof,

[0097]

[0098] wherein,

[0099] X, Y and U are each independently selected from CH and N, and when any one of them is CH, the hydrogen atom on the CH can be substituted by R1;

[0100] W is selected from CH and N;

[0101] Z is selected from CH2, NH, O and S;

[0102] L is selected from -CH=CH- and -C≡C-, and when n is greater than 1, -(L) n - is a chain formed by Ls that are the same as or different from each other;

[0103] Ring A is selected from C 6-8 cycloalkane, C 6-12 aryl ring, 6-10 membered heteroaryl ring and 6-8 membered heterocyclic ring;

[0104] Ring B is selected from C 6-12 aryl ring, C 4-8 cycloalkane, 5-10 membered heteroaryl ring and 4-8 membered heterocyclic ring;

[0105] M is a direct bond or a 4-8 membered heterocyclic ring;

[0106] R1s are each independently selected from: deuterium, tritium, hydroxyl, amino, halogen, nitro, cyano, -COOH, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -SO2-C 1-6 alkyl, acyl, ester, amide, sulfonamide, -NH-C 3-8 cycloalkyl, -N(C 1-6 alkyl)(C 3-8 cycloalkyl), -C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl group,

[0107] wherein, the -C 1-6 alkyl in R1 or the C as part of a group1-6 alkyl, -C 3-8 cycloalkyl or -C as part of a group 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, and 5- to 10-membered heteroaryl are each independently optionally substituted with one or more substituents, and each of the substituents is independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), and two substituents attached to the same C atom optionally together with the C atom to which they are attached form a 3- to 6-membered heterocycle;

[0108] R2 and R3 are each independently selected from: deuterium, tritium, hydroxy, amino, halogen, nitro, cyano, -COOH, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -SO2-C 1-6 alkyl, acyl, ester, amide, sulfonamide, -NH-C 3-8 cycloalkyl, -N(C 1-6 alkyl)(C 3-8 cycloalkyl), -C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered heteroaryl, and C 6-12 aryl, wherein the -C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered heteroaryl, and C 6-12 aryl are each independently optionally substituted with halogen, hydroxy, -OC 1-6 alkyl, -NHC 1-6 alkyl, or -N(C 1-6 alkyl)2;

[0109] wherein the -C 1-6 alkyl in R2 and R3 or C 1-6 alkyl as part of another group or a substituent is optionally substituted with one or more substituents each independently selected from the following: deuterium, tritium, halogen, hydroxy, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl);

[0110] n is an integer from 0 to 3;

[0111] p is an integer from 1 to 4;

[0112] q and t are each independently selected from integers from 0 to 5;

[0113] The heteroaryl and heterocyclic groups each independently contain 1, 2, 3, or 4 heteroatoms selected from N, S, or O; and

[0114] wherein each occurrence of a halogen is optionally in its isotopic form, and the carbon atoms and / or hydrogen atoms thereon in the substituents are optionally in their isotopic forms.

[0115] Embodiment 2: A compound according to the foregoing embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotopic variant thereof, wherein the acyl group, ester group, amide group, sulfonamide group is selected from -CO-C 1-6 alkyl, -C(O)OC 1-6 alkyl, -C(O)NH2, -C(O)NHC 1-6 alkyl, -C(O)N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl, -SO2NH2, -SO2NHC 1-6 alkyl, -SO2N(C 1-6 alkyl)2, -NHSO2-C 1-6 alkyl, -N(C 1-6 alkyl)-SO2-C 1-6 alkyl, and depending on the substituents in which they are located (such as R1, R2, or R3), the -C 1-6 alkyl is optionally substituted with the corresponding substituents as defined in the foregoing embodiments.

[0116] Embodiment 3: A compound according to any of the foregoing embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotopic variant thereof, wherein W is selected from CH and N; and Z is selected from O and S.

[0117] Embodiment 4: A compound according to any of the foregoing embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotopic variant thereof, wherein X, Y, and U are all CH, or one or two of X, Y, and U are N, and the rest are CH.

[0118] Embodiment 5: A compound according to any of the foregoing embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotopic variant thereof, wherein the fused ring containing X, Y, U, W, and Z is selected from:

[0119]

[0120] Embodiment 6: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is an integer from 1 to 2, preferably p is 1.

[0121] Embodiment 7.1: A compound according to any of the preceding embodiments 1-6, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 can be halogen, nitro or cyano, such as halogen and nitro, such as F, Br, NO2.

[0122] Embodiment 7.2: A compound according to any of the preceding embodiments 1-6, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 can be hydroxy or -O-C 1-6 alkyl, wherein the -C 1-6 alkyl is optionally substituted with one or more substituents, such as substituted with 1 or 2 substituents, the substituents being as defined in Embodiment 1, such as each independently selected from halogen, hydroxy, -O-(3-6 membered heterocyclic group), -O-(p-toluenesulfonyl), wherein the halogen is preferably F or 18F, and wherein the -O-(3-6 membered heterocyclic group) is preferably -O-THP.

[0123] Embodiment 7.3: A compound according to any of the preceding embodiments 1-6, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 can be amino, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl, -NHSO2-C 1-6 alkyl, -N(C 1-6 alkyl)-SO2-C 1-6 alkyl, -NH-C 3-8 cycloalkyl, -N(C 1-6 alkyl)(C 3-8 cycloalkyl), for example R1 can be -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl or -N(C 1-6 alkyl)-CO-C 1-6 alkyl,

[0124] wherein the -C 1-6The alkyl group is optionally substituted by one or more substituents, such as by 1 or 2 substituents, and the substituents are as defined in Embodiment 1, for example, each independently selected from halogen, hydroxy, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), wherein the halogen is preferably F or 18F, and the -O-(3- to 6-membered heterocyclic group) is preferably -O-THP.

[0125] Embodiment 7.4: A compound according to any of the preceding Embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 can be -COOH, -SO2-C 1-6 alkyl, -CO-C 1-6 alkyl, -C(O)OC 1-6 alkyl, -C(O)NH2, -C(O)NHC 1-6 alkyl, -C(O)N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl, -SO2NH2, -SO2NHC 1-6 alkyl, -SO2N(C 1-6 alkyl)2, -NHSO2-C 1-6 alkyl or -N(C 1-6 alkyl)-SO2-C 1-6 alkyl, for example, R1 can be -COOH or -NHCO-C 1-6 alkyl,

[0126] wherein the -C 1-6 alkyl is optionally substituted by one or more substituents, such as by 1 or 2 substituents, and the substituents are as defined in Embodiment 1, for example, each independently selected from halogen, hydroxy, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), wherein the halogen is preferably F or 18F, and the -O-(3- to 6-membered heterocyclic group) is preferably -O-THP.

[0127] Embodiment 7.5: A compound according to any of the preceding Embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 can be -C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 10-membered heteroaryl, for example, R1 can be -C 3-6 cycloalkyl, 5- to 8-membered heterocyclic group or 5- to 6-membered heteroaryl, for example, 5- to 8-membered heterocyclic group; each independently optionally substituted by one or more substituents, and the substituents are as defined in Embodiment 1, for example, each independently selected from -O-C 1-6 alkyl, -C 1-6Alkyl, halogen, hydroxy, oxo, -NHC 1-6 Alkyl, -N(C 1-6 alkyl)2.

[0128] Embodiment 7.6: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein each R1 is independently selected from: halogen, nitro, hydroxy, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl and 5- to 8-membered heterocyclic groups, wherein the -C 1-6 alkyl in R1 or the C 1-6 alkyl as part of a group and the 5- to 8-membered heterocyclic groups are each independently optionally substituted with one or more, for example 1 to 2, substituents selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), wherein the halogen in R1 is optionally in isotopic form, for example 18F.

[0129] Embodiment 7.7: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein each R1 is independently selected from halogen, nitro, hydroxy, -O-C 1-6 alkyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -O-C 1-6 alkyl substituted with O-(3- to 6-membered heterocyclic group) and / or -O-(p-toluenesulfonyl), -NHC 1-6 alkyl, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -NH-C 1-6 alkyl substituted with O-(3- to 6-membered heterocyclic group) and / or -O-(p-toluenesulfonyl), -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6alkyl and 5- or 6-membered heterocyclic group, where the 5- or 6-membered heterocyclic group is optionally substituted with one or more, for example 1-2 substituents, and each of the substituents is independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2,

[0130] Preferably, R1 is selected from -O-C 1-6 halo-hydroxyalkyl and -NHC 1-6 halo-hydroxyalkyl;

[0131] More preferably, R1 is selected from -O-C 1-6 halo-hydroxyalkyl;

[0132] wherein the halogen in R1 is optionally in isotopic form, for example 18F.

[0133] Embodiment 7.8: A compound according to any one of the preceding embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, where p is 2, and one of R1 is selected from H and halogen, and the other is selected from hydroxy, -O-C 1-6 alkyl, -O-methyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, NH2, -NHC 1-6 alkyl, -NHCH3, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -N(C 1-6 alkyl)2, -N(CH3)(C 1-6 alkyl), -N(C 1-6 alkyl)(C 1-6 haloalkyl), -N(C 1-6 alkyl)(C 1-6 halo-hydroxyalkyl), -N(CH3)(C 1-6 halo-hydroxyalkyl), -N(C 1-6 haloalkyl)(C 1-6 haloalkyl), -N(C 1-6 haloalkyl)(C 1-6 halo-hydroxyalkyl), -NH(C 3-8 cycloalkyl), -N(C 3-8 cycloalkyl)(C 1-6 alkyl), -N(C 3-8 cycloalkyl)(CH3), -N(C 3-8 cycloalkyl)(C 1-6 haloalkyl), -N(C 3-8 cycloalkyl)(C 1-6(halo-hydroxyalkyl), wherein the alkyl is optionally substituted with 1-3 -O-(3-6 membered heterocyclic group) or -O-(p-toluenesulfonyl).

[0134] Embodiment 7.9: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is 1 and R1 is selected from -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -N(C 1-6 alkyl)(C 1-6 haloalkyl), -N(C 1-6 alkyl)(C 1-6 halo-hydroxyalkyl), -N(C 1-6 haloalkyl)(C 1-6 haloalkyl), -N(C 1-6 haloalkyl)(C 1-6 halo-hydroxyalkyl), -N(C 3-8 cycloalkyl)(C 1-6 haloalkyl), -N(C 3-8 cycloalkyl)(C 1-6 halo-hydroxyalkyl).

[0135] Embodiment 7.10: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein each R1 is independently selected from: F, Br,

[0136] Embodiment 7.11: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein each R1 is independently selected from:

[0137] Embodiment 7.12: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein each R1 is independently selected from:

[0138] Embodiment 7.13: A compound according to any of the foregoing embodiments 1-6 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein R1 is selected from -O-C 1-6 halo-hydroxyalkyl and -NHC 1-6 halo-hydroxyalkyl, for example -O-C 1-6 halo-hydroxyalkyl, specifically selected from

[0139] Embodiment 7.14: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is 1 and R1 is located at the vicinal ring carbon atom of Y; or p is 2 and R1 is attached to the ring carbon atoms of Y and the vicinal carbon atom of Y respectively, or R1 is attached to the ring carbon atoms between X, X and Y respectively;

[0140] Preferably, p is 1 and R1 is located at the vicinal ring carbon atom of Y.

[0141] Embodiment 7.15: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein when both a hydroxyl group and a halogen are carried in R1, the hydroxyl group and the halogen are attached to different backbone atoms, such as C 1-6 different carbon atoms of an alkyl group, preferably attached to adjacent carbon atoms.

[0142] Embodiment 8.1: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein: n = 0.

[0143] Embodiment 8.2: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein: n is 1 and L is -CH=CH-, or L is -C≡C-.

[0144] Embodiment 8.3: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein n is 2 and (L)2 is -CH=CH-CH=CH-, -CH=CH-C≡C-, -C≡C-CH=CH- or -C≡C-C≡C-, preferably -CH=CH-CH=CH- or -CH=CH-C≡C-.

[0145] Embodiment 8.4: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein n is 3 and (L)3 is -CH=CH-CH=CH-CH=CH-, -CH=CH-CH=CH-C≡C-, -CH=CH-C≡C-CH=CH-, -C≡C-CH=CH-CH=CH-, -CH=CH-C≡C-C≡C-, -C≡C-CH=CH-C≡C- or -C≡C-C≡C-CH=CH-, preferably -CH=CH-CH=CH-CH=CH-, -CH=CH-CH=CH-C≡C-, -CH=CH-C≡C-CH=CH- or -CH=CH-C≡C-C≡C-.

[0146] Embodiment 9: A compound according to any of the preceding embodiments, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein formula (A) has the following sub-formula:

[0147]

[0148] Embodiment 10.1: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is a C 6-12 aromatic ring, such as a C 6-10 aromatic ring, such as a benzene ring.

[0149] Embodiment 10.2: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is a 6-10 membered heteroaromatic ring, such as a 6 membered heteroaromatic ring containing 1 or 2 nitrogen heteroatoms, such as

[0150] Embodiment 10.3: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is a 6-8 membered heterocyclic ring, such as a 6 membered heterocyclic ring containing 1 or 2 nitrogen heteroatoms, such as

[0151] Embodiment 10.4: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from a C 6-12 aromatic ring, a 6-10 membered heteroaromatic ring and a 6-8 membered heterocyclic ring, such as ring A is selected from a C 6-10 aromatic ring, a 6-8 membered heteroaromatic ring and a 6-8 membered heterocyclic ring, such as ring A is selected from a benzene ring and a 6 membered heteroaromatic ring or 6 membered heterocyclic ring containing 1 or 2 nitrogen heteroatoms, such as ring A is selected from a benzene ring and a 6 membered heteroaromatic ring containing 1 or 2 nitrogen heteroatoms, such as A is selected from Preferably ring A is selected from

[0152] Embodiment 10.5: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from a C 6-8 aromatic ring, a 6-8 membered heteroaromatic ring, such as ring A is selected from

[0153] Embodiment 10.6: A compound according to any of the preceding embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from a C 6-8An aromatic ring, a 6-8 membered heteroaromatic ring, for example ring A is selected from

[0154] Embodiment 10.7: A compound according to any of the foregoing embodiments 1-9 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from C 6-8 An aromatic ring, a 6-8 membered heteroaromatic ring, for example ring A is selected from

[0155] Embodiment 10.8: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein A is linked to the remainder of the molecule through a ring carbon atom, or when ring A is a heterocycle, it can also be linked to the remainder of the molecule through a ring heteroatom.

[0156] Embodiment 11.1: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is optionally substituted with 0-5 of said substituents R2, for example q is 0-4, 0-3, 0-2, 0-1, 1, preferably q is 0.

[0157] Embodiment 11.2: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is optionally substituted with 1 R2, and R2 is selected from halogen, nitro, cyano and halogen-substituted -C 1-6 alkyl, preferably selected from halogen, nitro and cyano, more preferably R2 is selected from halogen or its isotope, for example R2 is selected from F and 18F.

[0158] Embodiment 12.1: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from C 6-12 An aromatic ring, for example C 6-10 An aromatic ring, for example a benzene ring or a naphthalene ring.

[0159] Embodiment 12.2: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from 5-10 membered heteroaromatic rings, for example 5-8 membered heteroaryl groups, and further for example 5-6 membered heteroaromatic rings containing 1-3, for example 1-2 heteroatoms selected from nitrogen, oxygen and sulfur; for example B is selected from

[0160] Embodiment 12.3: A compound according to any of the foregoing embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from C 6-12Aromatic rings and 5- to 10-membered heteroaromatic rings, such as C 6-10 Aromatic rings and 5- to 8-membered heteroaromatic rings, for example ring B is selected from a benzene ring, a naphthalene ring, and a 5- to 6-membered heteroaromatic ring containing 1-3, for example 1-2, heteroatoms selected from nitrogen, oxygen, and sulfur, for example B is selected from

[0161] Preferably ring B is selected from

[0162] Embodiment 12.4: A compound according to any of the preceding embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from a benzene ring, a 5- to 8-membered heteroaromatic ring, for example ring B is selected from

[0163] Or for example ring B is selected from

[0164] Or ring B is a 6-membered heteroaromatic ring containing 2 or 3 heteroatoms selected from N, S or O, for example

[0165] Embodiment 13.1: A compound according to any of the preceding embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is optionally substituted with 0-5 of said substituents R3, for example t is 0-4, 0-3, 0-2, 0-1, 0, 1-2, preferably t is 1.

[0166] Embodiment 13.2: A compound according to any of the preceding embodiments or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is optionally substituted with 1 or 2 R3, and R3 is selected from hydroxy, amino, halogen, nitro, cyano, -C 1-6 alkyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -C 3-8 cycloalkyl, 5- to 8-membered heterocyclic group and C 6-10 aryl; wherein said -C 3-8 cycloalkyl, 5- to 8-membered heterocyclic group and C 6-10 aryl are preferably -C 3-6 cycloalkyl, 5- to 7-membered heterocyclic group and C6 aryl, each independently optionally substituted with halogen, hydroxy, -OC 1-6 alkyl, -NHC 1-6 alkyl or -N(C 1-6 alkyl)2;

[0167] wherein -C 1-6The alkyl group is optionally substituted by 1 or 2 substituents each independently selected from deuterium, tritium, halogen, hydroxyl, -O-(3-6-membered heterocyclic group), and -O-(p-toluenesulfonyl), such as deuterium, tritium, halogen, and hydroxyl, such as halogen or its isotope (e.g., F or 18F), and hydroxyl.

[0168] Embodiment 13.3: A compound or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotope variant thereof according to any of the preceding embodiments, wherein ring B is optionally substituted by 1 or 2 R3 substituents, and R3 is selected from hydroxyl, amino, halogen, nitro, cyano, -C 1-6 alkyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, 5-7-membered heterocyclic group, and C6 aryl; wherein the 5-7-membered heterocyclic group and C6 aryl are each independently optionally substituted by -NHC 1-6 alkyl or -N(C 1-6 alkyl)2;

[0169] wherein the -C 1-6 alkyl occurring in R3 is optionally substituted by 1 or 2 substituents each independently selected from halogen or its isotope (e.g., F or 18F), and hydroxyl.

[0170] Embodiment 13.4: A compound or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotope variant thereof according to any of the preceding embodiments, wherein ring B is optionally substituted by 1 or 2 R3 substituents, and R3 is selected from: hydroxyl, amino, F, Br, nitro, cyano, -O-CH3, -NHCH3, -N(CH3)2,

[0171] Embodiment 13.5: A compound or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotope variant thereof according to any of the preceding embodiments, wherein ring B is substituted by 1 substituent R3, and R3 is selected from -NHC 1-6 alkyl, such as -NHCH3.

[0172] Embodiment 14.1: A compound or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotope variant thereof according to any of the preceding embodiments, wherein M is a 6-8-membered nitrogen-containing heterocycle, such as a 6-membered nitrogen-containing heterocycle, such as

[0173] Embodiment 14.2: A compound or a stereoisomer, pharmaceutically acceptable salt, solvate, or stable isotope variant thereof according to any of the preceding embodiments, wherein n is 0, and M is a 6-membered nitrogen-containing heterocycle, such as

[0174] Embodiment 15: A compound according to any one of Embodiments 1-9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, having the following formula:

[0175]

[0176] wherein each group and variable is as defined in the corresponding foregoing embodiment, specifically

[0177] R1 is selected from halogen, nitro, hydroxy, -O-C 1-6 alkyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -O-C substituted with O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -NHC 1-6 alkyl, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -NH- substituted with O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl and 5-6 membered heterocyclic group, wherein the 5-6 membered heterocyclic group is optionally substituted with one or more substituents each independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, wherein the halogen in R1 is optionally in isotopic form, such as 18F;

[0178] Ring A is selected from C 6-12 aryl ring and 6-10 membered heteroaryl ring;

[0179] Ring B is selected from C 6-12 aryl ring and 5-10 membered heteroaryl ring;

[0180] R3 is selected from hydroxy, amino, halogen, nitro, cyano, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, 4-7 membered heterocyclic group;

[0181] t is an integer from 0-2,

[0182] The heteroaryl and heterocyclic groups each independently contain 1, 2 or 3 heteroatoms selected from N, S or O; and

[0183] Each occurrence of a halogen is optionally in its isotopic form, and the carbon atoms and / or the hydrogen atoms thereon in the substituent are optionally in their isotopic forms;

[0184] Embodiment 15.1: A compound according to Embodiment 15 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein:

[0185] R1 is selected from -O-C 1-6 halo-hydroxyalkyl and -NHC 1-6 halo-hydroxyalkyl, preferably R1 is selected from -O-C 1-6 halo-hydroxyalkyl;

[0186] Ring A is selected from a benzene ring and a 6-membered heteroaryl ring containing 1 or 2 nitrogen heteroatoms;

[0187] Ring B is selected from a benzene ring and a 5-6 membered heteroaryl ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur;

[0188] R3 is selected from -NHC 1-6 alkyl;

[0189] t is 1;

[0190] Each occurrence of a halogen is preferably F or 18F, and the carbon atoms and / or the hydrogen atoms thereon in the substituent are optionally in their isotopic forms.

[0191] Embodiment 15.2: A compound of the following formula or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof,

[0192]

[0193] wherein:

[0194] R1 is selected from hydroxy, -O-C 1-6 alkyl, -O-methyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, NH2, -NHC 1-6 alkyl, -NHCH3, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -N(C 1-6 alkyl)2, -N(CH3)(C 1-6 alkyl), -N(C 1-6 alkyl)(C 1-6 haloalkyl), -N(C 1-6 alkyl)(C 1-6(halo-hydroxyalkyl), -N(CH3)(C 1-6 (halo-hydroxyalkyl), -N(C 1-6 (haloalkyl)(C 1-6 (haloalkyl), -N(C 1-6 (haloalkyl)(C 1-6 (halo-hydroxyalkyl), -NH(C 3-8 (cycloalkyl), -N(C 3-8 (cycloalkyl)(C 1-6 (alkyl), -N(C 3-8 (cycloalkyl)(CH3), -N(C 3-8 (cycloalkyl)(C 1-6 (haloalkyl), -N(C 3-8 (cycloalkyl)(C 1-6 (halo-hydroxyalkyl), wherein the alkyl is optionally substituted by 1-3 -O-(3-6 membered heterocyclic group) or -O-(p-toluenesulfonyl);

[0195] Ring A is

[0196] Ring B is selected from C 6-12 aromatic ring, C 4-8 cycloalkane, 5-8 membered heteroaromatic ring and 4-8 membered heterocycle;

[0197] R3 is selected from deuterium, tritium, hydroxyl, amino, halogen group, nitro, cyano, carboxyl, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 haloalkylamino, methylsulfonyl, C 2-6 acyl, C 2-6 ester group, C 2-6 amide group, C 2-6 sulfonamide group, C 1-6 halo-hydroxyalkoxy, C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 5-8 membered heteroaryl.

[0198] t is an integer from 0 to 5;

[0199] wherein each occurrence of halogen is optionally in isotopic form.

[0200] Embodiment 16: A compound according to any of Embodiments 1-9 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, having the following formula:

[0201]

[0202] Each group and variable is as defined in the corresponding embodiments described above. Specifically,

[0203] R1 is selected from halogen, nitro, hydroxy, -O-C 1-6 alkyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -O-C substituted with O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -NHC 1-6 alkyl, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -NH-C substituted with O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl and 5-6 membered heterocyclic group, where the 5-6 membered heterocyclic group is optionally substituted with one or more substituents, and each of the substituents is independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2,

[0204] Ring A is selected from C 6-12 aryl ring and 6-10 membered heteroaryl ring;

[0205] Ring B is selected from C 6-12 aryl ring and 5-10 membered heteroaryl ring;

[0206] R3 is selected from hydroxy, amino, halogen, nitro, cyano, -C 1-6 alkyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, 5-7 membered heterocyclic group and C6 aryl; where the 5-7 membered heterocyclic group and C6 aryl are each independently optionally substituted with -NHC 1-6 alkyl or -N(C 1-6 alkyl)2, and the -C 1-6 alkyl in R3 is optionally substituted with one or two substituents each independently selected from halogen or halogen isotope (such as F or 18F) and hydroxy;

[0207] t is an integer from 0 to 2,

[0208] Each of the heteroaryl and heterocyclic groups independently contains 1, 2 or 3 heteroatoms selected from N, S or O; and

[0209] Each occurrence of a halogen is optionally in its isotopic form, and the carbon atoms and / or hydrogen atoms thereon in the substituents are optionally in their isotopic forms.

[0210] Embodiment 16.1: A compound according to Embodiment 16 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein

[0211] R1 is selected from -O-C 1-6 halo-hydroxyalkyl and -NHC 1-6 halo-hydroxyalkyl, preferably R1 is selected from -O-C 1-6 halo-hydroxyalkyl,

[0212] Ring A is selected from a benzene ring and a 6-membered heteroaryl ring containing 1 or 2 nitrogen heteroatoms;

[0213] Ring B is selected from a benzene ring and a 5-6 membered heteroaryl ring containing 1 or 2 heteroatoms selected from nitrogen, oxygen and sulfur;

[0214] R3 is selected from -NHC 1-6 alkyl;

[0215] t is 1;

[0216] Each occurrence of a halogen is preferably F or 18F, and the carbon atoms and / or hydrogen atoms thereon in the substituents are optionally in their isotopic forms.

[0217] Embodiment 16.2: A compound according to Embodiment 16 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof,

[0218]

[0219] wherein

[0220] R1 is selected from -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -N(C 1-6 alkyl)(C 1-6 haloalkyl), -N(C 1-6 alkyl)(C 1-6 halo-hydroxyalkyl), -N(C 1-6 haloalkyl)(C 1-6 haloalkyl), -N(C 1-6 haloalkyl)(C 1-6 halo-hydroxyalkyl), -N(C 3-8 cycloalkyl)(C 1-6(haloalkyl), -N(C 3-8 cycloalkyl)(C 1-6 (halohydroxyalkyl);

[0221] Ring A is

[0222] Ring B is selected from 6-membered heteroaryl rings;

[0223] R3 is selected from deuterium, tritium, hydroxy, amino, halogen, nitro, cyano, carboxy, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 hydroxyalkyl, C 2-6 alkenyl, C 2-6 alkynyl, -C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkylamino, C 1-6 haloalkylamino, methylsulfonyl, C 2-6 acyl, C 2-6 ester, C 2-6 amide, C 2-6 sulfonamide, C 1-6 halohydroxyalkoxy, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 8-membered heteroaryl.

[0224] t is an integer from 0 to 5;

[0225] Each occurrence of halogen is optionally in isotopic form.

[0226] Embodiment 16.3: A compound according to Embodiment 16 or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotopic variant thereof, wherein:

[0227] Ring A is

[0228] Ring B is selected from C 6-12 aryl ring, C 4-8 cycloalkane, 5- to 8-membered heteroaryl ring;

[0229] R1 is selected from: deuterium, tritium, hydroxy, C 1-6 alkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 8-membered heteroaryl, C 1-6 alkylamino, halogen, C 1-6 haloalkoxy, C 1-6 haloalkylamino, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 acyl, nitro, cyano, carboxy, C 2-6An ester group, an amino group, C 2-6 An amide group, -SO2CH3, C 2-6 A sulfonamide group; wherein the C 1-6 An alkyl group, C 1-6 An alkoxy group and C 1-6 The hydrogen atom on the alkylamino group may optionally be substituted by 1-3 groups selected from C 1-6 An alkoxy group, C 1-6 An alkyl group, a halogen, a hydroxyl group, OTHP, -O-(3-6 membered heterocyclic group), -O-(p-toluenesulfonyl group), and when two substituents are on the same C atom, they may together with the C atom to which they are attached form a 3-6 membered heterocyclic ring;

[0230] R3 is selected from: deuterium, tritium, a hydroxyl group, C 1-6 An alkoxy group, C 1-6 An alkyl group, C 3-8 A cycloalkyl group, a 3-8 membered heterocyclic group, a 3-8 membered heteroaryl group, C 1-6 An alkylamino group, a halogen, C 2-6 An alkenyl group, C 2-6 An alkynyl group, C 2-6 An acyl group, a nitro group, a cyano group, a carboxyl group, C 2-6 An ester group, an amino group, C 2-6 An amide group, -SO2CH3, C 2-6 A sulfonamide group; wherein the C 1-6 An alkoxy group and C 1-6 The alkylamino group is optionally substituted by 1-3 substituents selected from a halogen, a hydroxyl group, OTHP, -O-(3-6 membered heterocyclic group), -O-(p-toluenesulfonyl group);

[0231] Wherein the heteroaryl group, heteroaromatic ring, heterocyclic group and heterocycle each independently contain 1, 2, 3 or 4 heteroatoms selected from N, S or O;

[0232] The halogen includes its isotopes.

[0233] It should be noted that in any one of the above-mentioned Embodiment 15 to Embodiment 16.3, the specific examples of each variable are as exemplified in the corresponding foregoing embodiments.

[0234] Embodiment 17: A compound or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a stable isotope variant thereof according to any of the foregoing embodiments, wherein the isotope variant contains atoms selected from 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F and 36Cl; preferably contains 18F.

[0235] Embodiment 18: A compound or a stereoisomer, a pharmaceutically acceptable salt, a solvate or a stable isotope variant thereof, which is selected from:

[0236]

[0237]

[0238]

[0239]

[0240]

[0241] In more specific embodiments, the compounds of the present invention are selected from the following compounds and their stereoisomers, or their pharmaceutically acceptable salts or solvates:

[0242]

[0243]

[0244] In additional embodiments, some of the compounds of formula A of the present invention can also be defined as follows:

[0245]

[0246] or its pharmaceutically acceptable salt, or its solvate,

[0247] wherein X and Y are each independently selected from CH or N; and when said X or Y is CH, the hydrogen atom on said CH can be substituted by R b or R c substituted;

[0248] W is selected from CH, or N;

[0249] W and Z are each independently selected from CH2, NH, O or S;

[0250] Ring A is selected from the group consisting of: 4-8 membered saturated cycloalkanes, benzene rings, 5-8 membered heteroaryl rings, 4-8 membered heterocycles;

[0251] Ring B is respectively selected from the group consisting of: naphthalene rings, biphenyls, 4-8 membered saturated cycloalkanes, benzene rings, 5-8 membered heteroaryl rings, 4-8 membered heterocycles;

[0252] 0, 1, 2, 3, 4 or 5 hydrogens on said ring A or ring B are substituted by substituents selected from the group consisting of: deuterium, tritium, hydroxyl, C 1-6 alkoxy, C 1-6 halogenated hydroxyalkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic groups, 3-8 membered heteroaryl groups, C 1-6 alkylamino, halogen groups, C 1-6 haloalkyl, C1-6 Halogenated alkoxy, C 1-6 Halogenated alkylamino, C 2-6 Alkenyl, C 2-6 Alkynyl, C 2-6 Acyl, C 1-6 Hydroxyalkyl, nitro, cyano, carboxyl, C 2-6 Ester group, amino, C 2-6 Amide group, methylsulfonyl, C 2-6 Sulfonamide group;

[0253] R b and R c each independently selected from the group consisting of: deuterium, tritium, hydroxy, C 1-6 alkoxy, C 1-6 halogenated hydroxyalkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 5- to 8-membered heteroaryl, C 1-6 alkylamino, halogen, C 1-6 halogenated alkyl, C 1-6 halogenated alkoxy, C 1-6 halogenated alkylamino, C 1-6 halogenated hydroxyalkylamino, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 acyl, C 1-6 hydroxyalkyl, nitro, cyano, carboxyl, C 2-6 ester group, amino, C 2-6 amide group, methylsulfonyl, C 2-6 sulfonamide group, or JR'; wherein, the C 1-6 on the alkoxy, 0, 1, 2 or 3 hydrogen atoms can be replaced by R d and R e substituted, the R d and R e each independently selected from the group consisting of: C 1-6 alkoxy, halogen, hydroxy, TsO, OTHP, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), and when R d and R e substitute on the same C atom, R d , R e can together with the C atom to which it is attached form a 3- to 6-membered heterocyclic ring; J is O or NR a ; R a is selected from the group consisting of: hydrogen, C 1-6 alkyl, C 1-6 halogenated alkyl, 3- to 8-membered cycloalkyl; R' is selected from the group consisting of: hydrogen, methyl, C 1-6 alkyl, C 1-6 halogenated alkyl, C substituted with hydroxy 1-6Halogenated alkyl, wherein 0, 1, 2 or 3 hydrogen atoms on the C 1-6 alkyl can be substituted by -O-(3- to 6-membered heterocyclic group) or -O-(p-toluenesulfonyl);

[0254] Each of the heteroaryl, heteroaromatic ring, heterocyclic group and heterocycle independently contains 1, 2, 3 or 4 heteroatoms selected from N, S or O;

[0255] The halogen includes its isotopes.

[0256] In a preferred embodiment, ring A is selected from the group consisting of: 6- to 8-membered aromatic rings, 5- to 8-membered heteroaromatic rings; preferably, ring A is selected from the group consisting of:

[0257] In a preferred embodiment, ring B is selected from the group consisting of: benzene ring, 5- to 8-membered heteroaromatic rings; preferably, ring B is selected from the group consisting of:

[0258] In a preferred embodiment, the R b is hydrogen or halogen, and R c is W-R',

[0259] wherein W is O or NR a , and R a is selected from the group consisting of: hydrogen, C 1-6 alkyl, C 1-6 halogenated alkyl, 3- to 8-membered cycloalkyl;

[0260] R' is selected from the group consisting of: hydrogen, methyl, C 1-6 alkyl, C 1-6 halogenated alkyl, C 1-6 halogenated alkyl substituted by hydroxyl, wherein 0, 1, 2 or 3 hydrogen atoms on the C 1-6 alkyl can be substituted by -O-(3- to 6-membered heterocyclic group) or -O-(p-toluenesulfonyl);

[0261] Preferably, R c is selected from the group consisting of:

[0262] In a preferred embodiment, the compound includes a compound represented by formula I', or a pharmaceutically acceptable salt thereof, or a solvate thereof,

[0263]

[0264] wherein J is O or NR a ; and R a is selected from the group consisting of: hydrogen, C 1-6 alkyl, C 1-6 halogenated alkyl, 3- to 8-membered cycloalkyl;

[0265] R’ is selected from the group consisting of: hydrogen, methyl, C 1-6 alkyl, C 1-6 haloalkyl, C substituted with a hydroxyl group 1-6 haloalkyl, wherein 0, 1, 2 or 3 hydrogen atoms on the C 1-6 alkyl can be substituted with -O-(3-6 membered heterocyclic group) or -O-(p-toluenesulfonyl);

[0266] Ring A is

[0267] Ring B is defined as described in claim 1.

[0268] In a further embodiment, some compounds of formula A of the present invention can also be defined as follows:

[0269]

[0270] or a pharmaceutically acceptable salt thereof, or a solvate thereof,

[0271] wherein X and Y are each independently selected from CH or N; and when the X or Y is CH, the hydrogen atom on the CH can be substituted with R;

[0272] Z is selected from CH2, O, NH or S;

[0273] n is 1, 2 or 3;

[0274] Ring A is selected from the group consisting of: 6-8 membered saturated cycloalkanes, benzene ring, 5-8 membered heteroaryl rings, 6-8 membered heterocycles;

[0275] Ring B is selected from the group consisting of: naphthalene ring, biphenyl, benzene ring, 4-8 membered saturated cycloalkanes, 5 membered heteroaryl rings (such as pyridine), 7-8 membered heteroaryl rings;

[0276] R is selected from the group consisting of: deuterium, tritium, hydroxyl, C 1-6 alkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3-8 membered heterocyclic groups, 3-8 membered heteroaryl groups, C 1-6 alkylamino, halogen, C 1-6 haloalkoxy, C 1-6 haloalkylamino, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 acyl, nitro, cyano, carboxyl, C 2-6 ester group, amino, C 2-6 amide group, methylsulfonyl, C 2-6 sulfonamide group; wherein the C 1-6 alkyl, C 1-6 alkoxy and C 1-6Zero, one, two or three hydrogen atoms on the alkylamino group may be replaced by R b or R c , where the R b and R c are each independently selected from the following group: C 1-6 alkoxy, C 1-6 alkyl, halogen, hydroxy, TsO, OTHP, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), and when R b and R c are substituted on the same C atom, R b , R c and the C atom to which they are attached may together form a 3- to 6-membered heterocycle;

[0277] R a is selected from the following group: deuterium, tritium, hydroxy, C 1-6 alkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 3- to 8-membered heteroaryl, C 1-6 alkylamino, halogen, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 acyl, nitro, cyano, carboxyl, C 2-6 ester group, amino, C 2-6 amide group, methylsulfonyl, C 2-6 sulfonamide group; wherein, zero, one, two or three hydrogen atoms on the C 1-6 alkoxy and C 1-6 alkylamino may be replaced by substituents selected from the following group: halogen, hydroxy, TsO, OTHP, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl);

[0278] Unless otherwise specified, zero, one, two, three, four or five hydrogen atoms on each of the above groups may be further replaced by substituents selected from the following group: deuterium, tritium, hydroxy, halogen, C 1-6 alkoxy, C 1-6 haloalkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 3- to 8-membered heteroaryl, C 1-6 alkylamino, C 1-6 haloalkyl, C 1-6 haloalkoxy, C 1-6 haloalkylamino, C 1-6 haloalkoxyalkylamino, C 2-6 alkenyl, C 2-6 alkynyl, acyl, C 1-6 hydroxyalkyl, nitro, cyano, carboxyl, ester group, amino, amide group, methylsulfonyl, sulfonamide group;

[0279] Each of the heteroaryl group, heteroaromatic ring, heterocyclic group and heterocycle independently contains 1, 2, 3 or 4 heteroatoms selected from N, S or O;

[0280] The halogen includes its isotopes.

[0281] In another preferred example, the -O-(3-6 membered heterocyclic group) is -O-(tetrahydropyranyl).

[0282] In a preferred example, ring A is selected from the group consisting of: 6-8 membered aromatic rings, 5-8 membered heteroaromatic rings; preferably, ring A is selected from the group consisting of:

[0283] In a preferred example, ring B is selected from the group consisting of: benzene ring, 5-8 membered heteroaromatic rings; preferably, ring B is selected from the group consisting of:

[0284] In a preferred example, R is selected from the group consisting of:

[0285] In a preferred example, the compound includes a compound as shown in formula I', or a pharmaceutically acceptable salt thereof, or a solvate thereof,

[0286]

[0287] wherein, ring B, R and R a are defined as above.

[0288] In additional embodiments, some of the compounds of formula A of the present invention can also be defined as follows:

[0289]

[0290] or a pharmaceutically acceptable salt thereof, or a solvate thereof,

[0291] wherein, X and Y are each independently selected from CH or N;

[0292] Z is selected from CH2, O, NH or S;

[0293] n is 1, 2 or 3;

[0294] Ring A is selected from the group consisting of: 6-8 membered saturated cycloalkanes, benzene ring, 5-8 membered heteroaromatic rings, 6-8 membered heterocycles;

[0295] Ring B is a 6 membered heteroaromatic ring;

[0296] 0, 1, 2, 3, 4 or 5 hydrogens on the ring A or ring B are substituted with substituents selected from the group consisting of: deuterium, tritium, hydroxyl group, C 1-6 alkoxy group, C 1-6Halogenated hydroxyalkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 3- to 8-membered heteroaryl, C 1-6 alkylamino, halogen, C 1-6 halogenated alkyl, C 1-6 halogenated alkoxy, C 1-6 halogenated alkylamino, C 1-6 halogenated hydroxyalkylamino, C 2-6 alkenyl, C 2-6 alkynyl, acyl, C 1-6 hydroxyalkyl, nitro, cyano, carboxyl, C 2-6 ester group, amino, C 2-6 amide group, methylsulfonyl, C 2-6 sulfonamide group;

[0297] R is selected from the group consisting of: deuterium, tritium, hydroxy, C 1-6 alkoxy, C 1-6 alkyl, C 3-8 cycloalkyl, 3- to 8-membered heterocyclic group, 3- to 8-membered heteroaryl, C 1-6 alkylamino, halogen, C 1-6 halogenated alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 2-6 acyl, C 1-6 hydroxyalkyl, nitro, cyano, carboxyl, C 2-6 ester group, amino, C 2-6 amide group, methylsulfonyl, C 2-6 sulfonamide group; wherein, the C 1-6 alkoxy and C 1-6 alkylamino may have 0, 1, 2 or 3 hydrogen atoms substituted by R b or R c substituted, and the R b and R c are each independently selected from the group consisting of: C 1-6 alkoxy, halogen, hydroxy, TsO, OTHP, and when R b and R c are substituted on the same C atom, R b and R c may together with the C atom to which they are attached form a 3- to 6-membered heterocycle;

[0298] The heteroaryl, heteroaromatic ring, heterocyclic group and heterocycle each independently contain 1, 2, 3 or 4 heteroatoms selected from N, S or O;

[0299] The halogen includes its isotopes.

[0300] In a preferred embodiment, the ring A is selected from the group consisting of:

[0301] In a preferred embodiment, ring B is a 6-membered heteroaryl containing 2 or 3 heteroatoms selected from N, S or O;

[0302] Preferably, ring B is selected from the following group:

[0303] In a preferred embodiment, R is W-R',

[0304] wherein W is O or NR a ; and said R a is selected from the following group: C 1-6 alkyl, C 1-6 haloalkyl, 3- to 8-membered cycloalkyl;

[0305] R' is a C 1-6 haloalkyl substituted by a hydroxyl group, preferably selected from the following group:

[0306] In a preferred embodiment, the compound comprises a compound represented by formula I', or a pharmaceutically acceptable salt thereof, or a solvate thereof,

[0307]

[0308] wherein W is O or NR a , and R a is selected from the following group: C 1-6 alkyl, C 1-6 haloalkyl, 3- to 8-membered cycloalkyl;

[0309] R' is selected from the following group: C 1-6 haloalkyl substituted by a hydroxyl group, unsubstituted C 1-6 haloalkyl;

[0310] The definition of ring B is as described above.

[0311] It should be noted that for the compounds of formula (I), (II), and (III), in addition to the above definitions, each substituent may respectively have the meanings generally, specifically, or preferably defined for the corresponding substituent in formula (A) above, and the technical solutions formed by these substituents or any combination thereof are also covered by the present invention.

[0312] It should be understood that within the scope of the present invention, the above technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be repeated here one by one.

[0313] The main advantages of the present invention:

[0314] The compounds of the present application have novel structures, can specifically recognize α-synuclein aggregates, and can be used to prepare drugs for the treatment or diagnosis of neurodegenerative diseases related to α-synuclein aggregates and other misfolded proteins, which has great clinical significance.

[0315] Drug Compositions and Administration Methods

[0316] Since the compounds of the present invention can recognize α-synuclein aggregates and are used for the treatment or diagnosis of neurodegenerative diseases related to α-synuclein aggregates and other pathological aggregated proteins (such as Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Alzheimer's disease, amyotrophic lateral sclerosis, etc.). Therefore, the compounds of the present invention, their stereoisomers, their optical isomers, their pharmaceutically acceptable salts, their crystal forms, their isotope derivatives, their prodrugs, their metabolites, their solvates or their hydrates, as well as the pharmaceutical compositions containing the compounds of the present invention as the main active ingredient can be used for the treatment (stabilizing, alleviating or curing) or diagnosis of neurodegenerative diseases related to α-synuclein aggregates and other pathological aggregated proteins (such as Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Alzheimer's disease, amyotrophic lateral sclerosis, progressive supranuclear palsy, progressive muscular atrophy, etc.).

[0317] The pharmaceutical compositions of the present invention contain the compounds of the present invention within a safe and effective amount range and pharmaceutically acceptable excipients or carriers. Generally, the pharmaceutical composition contains 1 - 2000 mg of the compound of the present invention per dose, and more preferably, contains 10 - 200 mg of the compound of the present invention per dose. Preferably, the "per dose" is a capsule or a tablet.

[0318] Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as Tween )), wetting agents (such as sodium dodecyl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0319] The administration methods of the compounds or pharmaceutical compositions of the present invention include, but are not limited to, intravenous injection.

[0320] The compounds of the present invention can be administered alone or in combination with other pharmaceutically acceptable compounds.

[0321] Preparation Method

[0322] On the one hand, the present application provides a preparation method of a compound of formula (A-1), including the following methods:

[0323] Method 1: When R1 is not a hydroxyl group, an amino group, a C 1-6 halogenated hydroxyalkoxy group, a C 1-6 halogenated hydroxyalkylamino group, a C 1-6 halogenated alkoxy group or a C 1-6 halogenated alkylamino group,

[0324]

[0325] wherein the definitions of X, Y, U, Z, W, ring A and ring B are as described above;

[0326] Method 2: When R1 is an optionally substituted hydroxyl group or an optionally substituted amino group and the substituent is a C 1-6 alkyl group or a C 3-8 cycloalkyl group,

[0327]

[0328] wherein the definitions of X, Y, U, Z, W, ring A and ring B are as described above;

[0329] Method 3: When R1 is a substituted C 1-6 alkoxy group or a substituted C 1-6 alkylamino group, and the substituent is selected from halogen and OH,

[0330]

[0331] wherein R’ is selected from -C 1-6 alkyl group and C 1-6 halogenated alkyl group;

[0332] R” is selected from C 1-6 halogenated alkyl group;

[0333] R a is selected from H or -C 1-6 alkyl group;

[0334] the definitions of X, Y, U, Z, W, ring A and ring B are as described above;

[0335] Method 4: When R1 is a substituted C 1-6 alkoxy group or a substituted C 1-6 alkylamino group, and the substituent is selected from halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group),

[0336]

[0337] wherein R b is C 1-6an alkyl group, and 0, 1, 2, 3, 4, or 5 hydrogen atoms thereof are substituted with substituents selected from the group consisting of: halogen, -O-(p-toluenesulfonyl), -O-(3- to 6-membered heterocyclic group);

[0338] R a selected from H or -C 1-6 alkyl;

[0339] The definitions of X, Y, U, Z, W, ring A, and ring B are as described above.

[0340] In this regard, when the compound of formula (A) is defined by formula (I) as shown in the above specification, its corresponding preparation method can also be represented as follows:

[0341] Method 1. R b is not hydroxy, amino, C 1-6 halogenated hydroxyalkoxy, C 1-6 halogenated hydroxyalkylamino, C 1-6 halogenated alkoxy, or C 1-6 halogenated alkylamino

[0342]

[0343] R c , X, U, Z, W, ring A, and ring B are defined as above;

[0344] Method 2. When R b is hydroxy or amino

[0345]

[0346] wherein R3 is C1-6 alkyl or C1-6 cycloalkyl;

[0347] R c , X, U, Z, W, ring A, and ring B are defined as above;

[0348] Method 3. When R b is C 1-6 halogenated hydroxyalkoxy or C 1-6 halogenated hydroxyalkylamino

[0349]

[0350] wherein R1 is selected from the group consisting of: C 1-6 alkyl, C 1-6 haloalkyl, 3- to 8-membered cycloalkyl;

[0351] R2 is selected from the group consisting of: C 1-6 haloalkyl;

[0352] R3, R c, the definitions of J, X, U, Z, W, ring A and ring B are as described above;

[0353] Method 4.R b is C 1-6 haloalkoxy or C 1-6 haloalkylamino

[0354]

[0355] wherein, R4 is selected from the group consisting of: C 1-6 alkyl, and 0, 1, 2, 3, 4 or 5 hydrogen atoms on the C 1-6 alkoxy are substituted by substituents selected from the group consisting of: halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group);

[0356] R3, R c , the definitions of J, X, U, Z, W, ring A and ring B are as described above.

[0357] On the other hand, the present application provides a method for preparing a compound represented by formula (A-2), including the following methods:

[0358] Method 1: When R1 is not hydroxy, C 1-6 hydroxyalkoxy, C 1-6 halohydroxyalkoxy or C 1-6 halohydroxyalkylamino

[0359]

[0360] wherein, the definitions of X, Y, U, W, Z, n, ring A and ring B are as described above;

[0361] Method 2: When R1 is hydroxy

[0362]

[0363] wherein, R c is selected from: C 1-6 haloalkyl; the definitions of X, Y, U, W, Z, n, ring A and ring B are as described above;

[0364] Method 3: When R1 is C 1-6 halohydroxyalkoxy or C 1-6 halohydroxyalkylamino

[0365]

[0366] wherein, R' is selected from the group consisting of: C 1-6 alkyl and C 1-6 haloalkyl;

[0367] R'' is selected from the group consisting of: C1-6 Halogenated alkyl;

[0368] R a Selected from H or -C 1-6 Alkyl;

[0369] The definitions of X, Y, U, W, Z, n, ring A and ring B are as described above;

[0370] Method 4: R1 is C 1-6 Alkoxy, and the C 1-6 Alkoxy is optionally substituted by 1-5 substituents selected from the group consisting of halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group),

[0371]

[0372] wherein the R b Selected from the group consisting of: C 1-6 Alkyl, and 0, 1, 2, 3, 4 or 5 hydrogen atoms thereof are substituted by substituents selected from the group consisting of halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group);

[0373] The definitions of W, U, X, Y, Z, n, ring A and ring B are as described above.

[0374] In this regard, when the compound of formula (A) is defined by formula (II) or formula (III) shown in the above specification, its corresponding preparation method can also be represented as follows:

[0375] Method 1: When R is not hydroxy, C 1-6 Hydroxyalkoxy, C 1-6 Halogenated hydroxyalkoxy or C 1-6 Halogenated hydroxyalkylamino,

[0376]

[0377] wherein the definitions of X, Y, Z, n, ring A and ring B are as described above;

[0378] Method 2: When R is hydroxy,

[0379]

[0380] wherein R' is selected from: C 1-6 Halogenated alkyl; the definitions of X, Y, Z, n, ring A and ring B are as described above;

[0381] Method 3: When R is C 1-6 Halogenated hydroxyalkoxy or C 1-6 Halogenated hydroxyalkylamino

[0382]

[0383] Wherein, W is O, S or NH;

[0384] R1 is selected from the group consisting of: C 1-6 alkyl, 1-6 haloalkyl, 3-8 membered cycloalkyl;

[0385] R2 is selected from the group consisting of: C 1-6 haloalkyl;

[0386] The definitions of X, Y, Z, n, ring A and ring B are as described above;

[0387] Method 4: R is C 1-6 alkoxy, and 0, 1, 2, 3, 4 or 5 hydrogen atoms on the C 1-6 alkoxy are substituted by substituents selected from the group consisting of: halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group),

[0388]

[0389] Wherein, R4 is selected from the group consisting of: C 1-6 alkyl, and 0, 1, 2, 3, 4 or 5 hydrogen atoms on the C 1-6 alkoxy are substituted by substituents selected from the group consisting of: halogen, -O-(p-toluenesulfonyl), -O-(3-6 membered heterocyclic group);

[0390] The definitions of W, X, Y, Z, n, ring A and ring B are as described above.

[0391] Abbreviations

[0392] Ts: p-toluenesulfonyl

[0393] THP: tetrahydropyranyl

[0394] Ex-vivo: ex-vivo experiment

[0395] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following embodiments are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are weight percentages and weight parts. The known starting materials of the present invention can be used or synthesized according to methods known in the art, or purchased as commercial products from multiple reagent companies. The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry.

[0396] Example I-1: Preparation of Compound I-1

[0397]

[0398] Synthetic route of Compound I-1:

[0399]

[0400] First step: Preparation of intermediate Ia-1

[0401] Dissolve 6-methoxy-2-bromobenzothiazole (537 mg, 3 mmol) and 2-fluoro-5-pyridineboronic acid (423 mg, 3 mmol) in 15 mL of solvent (1,4-dioxane:H2O = 4:1). Add K2CO3 (1.24 g, 9 mmol) and Pd(dppf)Cl2 (219 mg, 0.3 mmol), protect with N2, and react at 80 °C for 8 h. Subsequently, pour the reaction solution into 100 mL of saturated NH4Cl solution, extract three times with ethyl acetate, wash three times with brine, dry over anhydrous sodium sulfate, and then purify by column chromatography to obtain Ia-1 as a pale yellow solid with a yield of 49%, ESI-MS (positive): 261.0 [M+1] + .

[0402] Second step: Preparation of intermediate Ib-1

[0403] Add 1-tert-butoxycarbonylpiperazine (373 mg, 2 mmol), 4-bromo-2-(methylamino)pyridine (357 mg, 2 mmol), Cs2CO3 (1.96 g, 6 mmol), Pd2(dba)3 (36 mg, 0.04 mmol), and XantPhos (70 mg, 0.12 mmol) into a round-bottom flask, add 10 mL of THF, protect with N2, and react at 80 °C for 12 h. After the reaction is completed, remove THF by vacuum distillation under reduced pressure, add 20 mL of ethyl acetate to dissolve the product, wash three times with brine, dry over anhydrous sodium sulfate, and then purify by column chromatography. The purified product is dissolved in 20 mL of dichloromethane, add 2 mL of trifluoroacetic acid, and stir at room temperature for 2 h. Remove the solvent by vacuum distillation under reduced pressure to obtain intermediate Ib-1 as a yellow solid with a yield of 57%, ESI-MS (positive): 193.1 [M+1] + .

[0404] Third step: Preparation of intermediate Ic-1

[0405] Dissolve intermediate Ia-1 (0.2 mmol) and Ib-1 (0.2 mmol) in 1 mL of NMP, add DIPEA (200 μL, 1 mmol), protect with N2, and react by microwave at 180 °C for 4 h. After the reaction is completed, add 10 mL of water, extract with 20 mL of ethyl acetate, wash three times with brine, dry over anhydrous sodium sulfate, and then purify by column chromatography to obtain Ic-1 as a pale yellow solid with a yield of 50%, ESI-MS (positive): 432.8 [M+1] + 。

[0406] Step 4: Prepare intermediate Id-1

[0407] Dissolve intermediate Ic-1 (0.1 mmol) in 0.3 mL of dichloromethane, cool the solution to -78 °C, and then dropwise add BBr3 (0.3 mL, 1 M in DCM), and react at room temperature for 12 h. After the reaction is completed, pour the reaction solution into 5 mL of water, extract with 20 mL of ethyl acetate, wash three times with brine, dry over anhydrous sodium sulfate, and evaporate to dryness to obtain intermediate Id-1 as a yellow solid with a yield of 81%, ESI-MS (positive): 418.9 [M+1] + 。 1 1H NMR (400 MHz, DMSO-d6) δ 12.14–11.98 (m, 1H), 9.82 (s, 1H), 8.74 (d, J = 2.5 Hz, 1H), 8.13 (dd, J = 9.0, 2.5 Hz, 1H), 7.78 (d, J = 8.8 Hz, 1H), 7.66 (t, J = 6.3 Hz, 1H), 7.56 (d, J = 5.2 Hz, 1H), 7.38 (d, J = 2.4 Hz, 1H), 7.01–6.92 (m, 2H), 6.61 (dd, J = 7.6, 2.5 Hz, 1H), 5.92 (s, 1H), 3.88–3.79 (m, 4H), 3.76–3.71 (m, 4H), 2.86 (d, J = 4.8 Hz, 3H).

[0408] Step 5: Prepare product I-1

[0409] Dissolve intermediate Id-1 (0.05 mmol) in 0.5 mL of DMF, add K2CO3 (20 mg, 0.15 mmol) and epifluorohydrin (8 μL, 0.1 mmol), and react the solution at 80 °C for 4 h. After the reaction is completed, purify the reaction solution by column chromatography to obtain the final product I-1 as a pale yellow solid with a yield of 61%, ESI-MS (positive): 495.0 [M+1] + 。 11H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 8.78 (d, J = 2.6 Hz, 1H), 8.16 (dd, J = 9.0, 2.6 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.75 - 7.63 (m, 2H), 7.58 (d, J = 5.6 Hz, 1H), 7.12 (dd, J = 8.9, 2.6 Hz, 1H), 6.99 (d, J = 9.1 Hz, 1H), 6.61 (dd, J = 7.7, 2.5 Hz, 1H), 5.92 (s, 1H), 4.58 (dd, J = 9.5, 4.3 Hz, 1H), 4.52–4.39 (m, 1H), 4.13 - 4.01 (m, 3H), 3.88 - 3.83 (m, 4H), 3.76 - 3.72 (m, 4H), 2.86 (d, J = 4.7 Hz, 3H).

[0410] Example I-2: Preparation of Compound I-2

[0411]

[0412] Using the synthesis method of Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 5-bromo-2-(methylamino)pyridine. Compound I-2 was obtained as a yellow solid with a yield of 21%, ESI-MS (positive): 495.0 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 8.77 (d, J = 2.5 Hz, 1H), 8.42 (s, 1H), 8.14 (dd, J = 9.0, 2.5 Hz, 1H), 8.01 (dd, J = 9.8, 2.7 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.71 (d, J = 2.6 Hz, 1H), 7.26 (d, J = 2.7 Hz, 1H), 7.15 - 6.99 (m, 3H), 4.62–4.58 (m, 1H), 4.50 - 4.41 (m, 1H), 4.14 - 4.02 (m, 3H), 3.82 (t, J = 5.0 Hz, 4H), 3.14 (t, J = 5.0 Hz, 4H), 2.91 (s, 3H).

[0413] Example I-3: Preparation of Compound I-3

[0414]

[0415] Using the synthesis method of Example I-1, only replace 2-fluoro-5-pyridineboronic acid with 3-fluoro-6-pyridineboronic acid; replace 4-bromo-2-(methylamino)pyridine with 2-bromopyridine. The obtained compound I-3 is a pale yellow solid with a yield of 5%. ESI-MS (positive): 466.1 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (d, J = 2.9 Hz, 1H), 8.15 - 8.05 (m, 2H), 7.89 (d, J = 8.9 Hz, 1H), 7.82 - 7.75 (m, 1H), 7.70 (d, J = 2.6 Hz, 1H), 7.53 (dd, J = 9.0, 2.9 Hz, 1H), 7.18 - 7.08 (m, 2H), 6.82 (t, J = 6.3 Hz, 1H), 4.58 (dd, J = 9.6, 4.3 Hz, 1H), 4.46 (dd, J = 9.9, 4.8 Hz, 1H), 4.14 - 4.01 (m, 3H), 3.78 - 3.74 (m, 4H), 3.59 - 3.55 (m, 4H).

[0416] Example I-4: Preparation of Compound I-4

[0417]

[0418] Using the synthesis method of Example I-1, only replace 4-bromo-2-(methylamino)pyridine with 4-bromo-2-(dimethylamino)pyridine. The obtained compound I-4 is a yellow solid with a yield of 11%. ESI-MS (positive): 509.0 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 11.93 (s, 1H), 8.78 (d, J = 2.5 Hz, 1H), 8.16 (dd, J = 8.9, 2.5 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.75 - 7.66 (m, 2H), 7.12 (dd, J = 8.9, 2.5 Hz, 1H), 7.00 (d, J = 9.0 Hz, 1H), 6.66 (dd, J = 7.6, 2.3 Hz, 1H), 5.96 (d, J = 2.3 Hz, 1H), 4.58 (dd, J = 9.6, 4.3 Hz, 1H), 4.51 - 4.41 (m, 1H), 4.15 – 4.02 (m, 3H), 3.87 - 3.82 (m, 4H), 3.81 - 3.76 (m, 4H), 3.13 (s, 6H).

[0419] Example I-5: Preparation of Compound I-5

[0420]

[0421] Using the synthesis method of Example I-1, only replace 4-bromo-2-(methylamino)pyridine with 2-bromopyrazine. The obtained compound I-5 is a brown solid with a yield of 15%. ESI-MS(positive): 467.2[M+1] + 。 1 H NMR(400MHz, DMSO-d6) δ8.77(d, J = 2.6Hz, 1H), 8.38(s, 1H), 8.19 - 8.11(m, 2H), 7.92 - 7.84(m, 2H), 7.71(d, J = 2.6Hz, 1H), 7.12(dd, J = 8.9, 2.6Hz, 1H), 7.04(d, J = 9.1Hz, 1H), 4.58(dd, J = 9.5, 4.2Hz, 1H), 4.51 - 4.38(m, 1H), 4.16 - 4.02(m, 3H), 3.83 - 3.79(m, 4H), 3.75 - 3.71(m, 4H).

[0422] Example I-6: Preparation of Compound I-6

[0423]

[0424] Using the synthesis method of Example I-1, only replace 4-bromo-2-(methylamino)pyridine with 5-bromo-3-(methylamino)pyridine. The obtained compound I-6 is a yellow solid with a yield of 22%. ESI-MS(positive): 495.2[M+1] + 。 1 H NMR(400MHz, DMSO-d6) δ8.78(d, J = 2.5Hz, 1H), 8.16(dd, J = 8.9, 2.7Hz, 1H), 7.87(d, J = 8.9Hz, 1H), 7.80 - 7.75(m, 1H), 7.71(d, J = 2.6Hz, 1H), 7.61 - 7.43(m, 1H), 7.13(dd, J = 8.9, 2.6Hz, 1H), 7.09 - 7.04(m, 1H), 6.98 - 6.90(m, 1H), 4.58(dd, J = 9.5, 4.3Hz, 1H), 4.50 - 4.39(m, 1H), 4.13 - 4.02(m, 3H), 3.84(t, J = 4.8Hz, 4H), 3.38 - 3.30(m, 4H), 2.80(s, 3H).

[0425] Example I-7: Preparation of Compound I-7

[0426]

[0427] Using the synthesis method of Example I-1, only replacing 4-bromo-2-(methylamino)pyridine with 5-bromo-pyrimidine. The obtained compound I-7 is a brown solid with a yield of 9%. ESI-MS (positive): 467.0 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.81 - 8.75 (m, 1H), 8.66 - 8.53 (m, 3H), 8.19 - 8.11 (m, 1H), 7.92 - 7.83 (m, 1H), 7.75 - 7.67 (m, 1H), 7.19 - 7.03 (m, 2H), 4.66 - 4.53 (m, 1H), 4.51 - 4.42 (m, 1H), 4.09 - 4.03 (m, 3H), 3.87 - 3.81 (m, 4H), 3.48 - 3.40 (m, 4H).

[0428] Example I-8: Preparation of Compound I-8

[0429]

[0430] Using the synthesis method of Example I-1, only replacing 2-fluoro-5-pyridineboronic acid with 3-fluoro-6-pyridineboronic acid. The obtained compound I-8 is a yellow solid with a yield of 20%. ESI-MS (positive): 495.1 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 12.04 (s, 1H), 8.42 (d, J = 2.9 Hz, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.70–7.63 (m, 2H), 7.55 (d, J = 5.4 Hz, 1H), 7.49 (dd, J = 8.9, 2.9 Hz, 1H), 7.12 (dd, J = 8.9, 2.6 Hz, 1H), 6.65 (dd, J = 7.5, 2.5 Hz, 1H), 5.98–5.90 (m, 1H), 4.63–4.52 (m, 1H), 4.50–4.40 (m, 1H), 4.14–4.02 (m, 3H), 3.76 (t, J = 5.0 Hz, 4H), 3.58 (t, J = 5.3 Hz, 4H), 2.87 (d, J = 4.8 Hz, 3H).

[0431] Example I-9: Preparation of Compound I-9

[0432]

[0433] Using the synthesis method of Intermediate Id-1 in Example I-1, only replacing 4-bromo-2-(methylamino)pyridine with 4-bromo-2-fluoropyridine. The obtained Compound I-9 is a yellow solid with a yield of 6%. ESI-MS (positive): 408.0 [M+1] + 。 1 HNMR(400MHz,DMSO-d6)δ9.81(s,1H),8.73(d,J=2.5Hz,1H),8.11(dd,J=9.0,2.5Hz,1H),7.84(d,J=6.1Hz,1H),7.78(d,J=8.8Hz,1H),7.38(d,J=2.4Hz,1H),7.01(d,J=9.0Hz,1H),6.95(dd,J=8.8,2.5Hz,1H),6.88–6.78(m,1H),6.52(s,1H),3.83–3.77(m,4H),3.59–3.54(m,4H).

[0434] Example I-10: Preparation of Compound I-10

[0435]

[0436] Using the synthesis method of Intermediate Id-1 in Example I-1, only replacing 4-bromo-2-(methylamino)pyridine with 5-bromo-2-fluoropyridine. The obtained Compound I-10 is a yellow solid with a yield of 11%. ESI-MS (positive): 408.0 [M+1] + 。 1 HNMR(400MHz,DMSO-d6)δ9.81(s,1H),8.73(d,J=2.5Hz,1H),8.11(dd,J=9.0,2.5Hz,1H),7.92–7.86(m,1H),7.78(d,J=8.7Hz,1H),7.71–7.64(m,1H),7.38(d,J=2.5Hz,1H),7.10–7.03(m,2H),6.96(dd,J=8.8,2.5Hz,1H),3.81(t,J=5.2Hz,4H),3.28(t,J=5.1Hz,4H).

[0437] Example I-11: Preparation of Compound I-11

[0438]

[0439] Using the synthesis method of Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 4-bromo-pyridine. The obtained compound I-11 is a yellow solid with a yield of 21%. ESI-MS (positive): 466.0 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.78 (d, J = 2.4 Hz, 1H), 8.33–8.27 (m, 2H), 8.17 (dd, J = 9.0, 2.5 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.71 (d, J = 2.5 Hz, 1H), 7.32–7.18 (m, 2H), 7.13 (dd, J = 8.9, 2.5 Hz, 1H), 7.04–6.96 (m, 1H), 4.63–4.53 (m, 1H), 4.52–4.41 (m, 1H), 4.16–4.01 (m, 3H), 3.89 (s, 8H).

[0440] Example I-12: Preparation of Compound I-12

[0441]

[0442] Using the synthesis method of Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 2-bromo-pyridine. The obtained compound I-12 is a yellow solid with a yield of 13%. ESI-MS (positive): 466.0 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.80–8.73 (m, 1H), 8.16 (dd, J = 9.0, 2.6 Hz, 1H), 8.10 (d, 1H), 7.91–7.81 (m, 2H), 7.71 (d, J = 2.6 Hz, 1H), 7.21 (d, J = 8.9 Hz, 1H), 7.12 (dd, J = 8.9, 2.6 Hz, 1H), 7.03 (d, J = 9.1 Hz, 1H), 6.86 (t, J = 6.3 Hz, 1H), 4.64–4.52 (m, 1H), 4.51–4.40 (m, 1H), 4.15–4.01 (m, 3H), 3.93–3.73 (m, 8H).

[0443] Example I-13: Preparation of Compound I-13

[0444]

[0445] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine is replaced with 2-bromo-5-hydroxypyridine. Compound I-13 was obtained as a yellow solid with a yield of 23%. ESI-MS(positive): 405.9[M+1] + 。 1 H NMR(400MHz, DMSO-d6)δ10.01(s,1H), 8.74(d,J=2.5Hz,1H), 8.17–8.09(m,1H), 7.78(d,J=8.8Hz,1H), 7.66(d,J=2.8Hz,1H), 7.45(d,J=9.4Hz,1H), 7.40–7.36(m,1H), 7.13(d,J=9.4Hz,1H), 7.04(d,J=9.1Hz,1H), 6.98–6.94(m,1H), 3.85–3.78(m,4H), 3.63–3.57(m,4H).

[0446] Example I-14: Preparation of Compound I-14

[0447]

[0448] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine is replaced with 2-bromo-5-hydroxypyridine. Compound I-14 was obtained as a yellow solid with a yield of 2%. ESI-MS(positive): 484.1[M+1] + 。 1 H NMR(400MHz, DMSO-d6)δ10.69(s,1H), 8.77(s,1H), 8.18–8.09(m,1H), 7.81(d,J=8.8Hz,1H), 7.67(d,J=2.9Hz,1H), 7.42(d,J=9.4Hz,1H), 7.15(d,J=8.9Hz,1H), 7.10(d,J=9.4Hz,1H), 7.04(d,J=9.1Hz,1H), 3.83(t,J=5.3Hz,4H), 3.66–3.57(m,4H).

[0449] Example I-15: Preparation of Compound I-15

[0450]

[0451] Using the synthetic method of intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine is replaced with 5-bromo-2-dimethylaminopyridine. Compound I-15 was obtained as a dark yellow solid with a yield of 8%. ESI-MS (positive): 433.0 [M+1] + 。 1 HNMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 8.67 (d, J = 2.5 Hz, 1H), 8.05 (dd, J = 9.0, 2.5 Hz, 1H), 7.96 (d, J = 9.7 Hz, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.33–7.26 (m, 2H), 7.14 (d, J = 9.9 Hz, 1H), 7.00 (d, J = 9.1 Hz, 1H), 6.89 (dd, J = 8.8, 2.5 Hz, 1H), 3.78–3.71 (m, 4H), 3.15–3.05 (m, 10H).

[0452] Example I-16: Preparation of Compound I-16

[0453]

[0454] Using the synthetic method of intermediate Id-1 in Example I-1, except that 2-fluoro-5-pyridineboronic acid is replaced with 3-fluoro-6-pyridineboronic acid, and 4-bromo-2-(methylamino)pyridine is replaced with 2-bromo-pyridine. Compound I-16 was obtained as a dark yellow solid with a yield of 14%. ESI-MS (positive): 389.9 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 9.83 (s, 1H), 8.43 (d, J = 2.8 Hz, 1H), 8.14–8.05 (m, 2H), 7.90–7.77 (m, 2H), 7.52 (dd, J = 8.9, 2.9 Hz, 1H), 7.36 (d, J = 2.4 Hz, 1H), 7.22 (d, J = 9.0 Hz, 1H), 6.96 (dd, J = 8.8, 2.5 Hz, 1H), 6.85 (t, J = 6.4 Hz, 1H), 3.82–3.75 (m, 4H), 3.60–3.54 (m, 4H).

[0455] Example I-17:: Preparation of Compound I-17

[0456]

[0457] Using the synthetic method of intermediate Id-1 in Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 4-bromo-2-dimethylaminopyridine. Compound I-17 was obtained as a dark yellow solid with a yield of 11%. ESI-MS (positive): 433.0 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 11.91 (s, 1H), 9.83 (s, 1H), 8.74 (d, J = 2.5 Hz, 1H), 8.13 (dd, J = 9.0, 2.6 Hz, 1H), 7.78 (d, J = 8.7 Hz, 1H), 7.70–7.64 (m, 1H), 7.38 (d, J = 2.5 Hz, 1H), 7.02–6.90 (m, 2H), 6.66 (dd, J = 7.4, 2.4 Hz, 1H), 5.96 (s, 1H), 3.87–3.80 (m, 4H), 3.79–3.76 (m, 4H), 3.13 (s, 6H).

[0458] Example I-18: Preparation of Compound I-18

[0459]

[0460] Using the synthetic method of intermediate Ic-1 in Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 2-bromo-5-hydroxypyridine. Compound I-18 was obtained as a yellow solid with a yield of 35%. ESI-MS (positive): 420.1 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.15 (d, J = 8.9 Hz, 1H), 7.91–7.83 (m, 1H), 7.72–7.64 (m, 2H), 7.43 (d, J = 9.3 Hz, 1H), 7.11 (d, J = 9.2 Hz, 2H), 7.05 (d, J = 9.2 Hz, 1H), 3.85 (s, 3H), 3.84–3.80 (m, 4H), 3.61–3.56 (m, 4H).

[0461] Example I-19: Preparation of Compound I-19

[0462]

[0463] Using the synthetic method of Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 3-bromopyridine. Compound I-19 was obtained as a yellow solid with a yield of 21%. ESI-MS (positive): 466.1 [M+1] + 。1 1H NMR (400 MHz, DMSO-d6) δ 8.81–8.74 (m, 1H), 8.48 (s, 1H), 8.23–8.11 (m, 2H), 8.01 (d, J = 8.7 Hz, 1H), 7.91–7.83 (m, 1H), 7.82–7.73 (m, 1H), 7.73–7.68 (m, 1H), 7.21–7.01 (m, 2H), 4.64–4.40 (m, 2H), 4.13–4.01 (m, 3H), 3.86 (s, 4H), 3.56 (s, 4H).

[0464] Example I-20: Preparation of Compound I-20

[0465]

[0466] Using the synthetic method of Intermediate Id-1 in Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 5-bromo-thiazole. Compound I-20 was obtained as a yellow solid with a yield of 17%. ESI-MS (positive): 396.2 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 9.80 (s, 1H), 8.77–8.69 (m, 1H), 8.45 (s, 1H), 8.11 (dd, J = 9.0, 2.5 Hz, 1H), 7.78 (d, J = 8.8 Hz, 1H), 7.38 (d, J = 2.5 Hz, 1H), 7.19 (s, 1H), 7.06 (d, J = 9.0 Hz, 1H), 6.95 (dd, J = 8.8, 2.5 Hz, 1H), 3.81 (t, J = 5.0 Hz, 4H), 3.21 (t, J = 5.0 Hz, 4H).

[0467] Example I-21: Preparation of Compound I-21

[0468]

[0469] Using the synthetic method of Intermediate Ic-1 in Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 5-bromo-thiazole. Compound I-21 was obtained as a yellow solid with a yield of 35%. ESI-MS (positive): 410.1 [M+1] + . 11H NMR (400 MHz, Chloroform-d) δ 8.72 (s, 1H), 8.28 (s, 1H), 8.13 (d, J = 9.0 Hz, 1H), 7.82 (d, J = 9.0 Hz, 1H), 7.18 (s, 1H), 7.10 (s, 1H), 7.03–6.95 (m, 1H), 6.70 (d, J = 9.0 Hz, 1H), 3.84–3.75 (m, 8H), 3.23–3.12 (m, 3H).

[0470] Example I-22: Preparation of Compound I-22

[0471]

[0472] Using the synthesis method of Example I-1, except replacing 4-bromo-2-(methylamino)pyridine with 3-pyridine and replacing epifluorohydrin with 1-bromo-2-fluoroethane. Compound I-22 was obtained as a yellow solid with a yield of 25%. ESI-MS (positive): 436.1 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 8.79 (s, 1H), 8.46 (s, 1H), 8.24–8.11 (m, 2H), 7.97–7.82 (m, 2H), 7.75–7.60 (m, 2H), 7.18–7.01 (m, 2H), 4.92–4.56 (m, 2H), 4.43–4.10 (m, 2H), 3.88–3.82 (m, 4H), 3.56–3.45 (m, 4H).

[0473] Example I-23: Preparation of Compound I-23

[0474]

[0475] Using the synthesis method of Intermediate Id-1 in Example I-1, except replacing 2-fluoro-5-pyridineboronic acid with 4-fluorobenzeneboronic acid and replacing 4-bromo-2-(methylamino)pyridine with 2-pyridine. Compound I-23 was obtained as a yellow solid with a yield of 19%. ESI-MS (positive): 388.9 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 9.78 (s, 1H), 8.11 (dd, J = 5.7, 1.9 Hz, 1H), 7.94–7.78 (m, 3H), 7.76 (d, J = 8.8 Hz, 1H), 7.36 (d, J = 2.4 Hz, 1H), 7.20 (d, J = 8.8 Hz, 1H), 7.10 (d, J = 8.9 Hz, 2H), 6.94 (dd, J = 8.8, 2.4 Hz, 1H), 6.84 (t, J = 6.4 Hz, 1H), 3.76 (t, J = 5.0 Hz, 4H), 3.50 (t, J = 5.3 Hz, 4H).

[0476] Example I-24: Preparation of Compound I-24

[0477]

[0478] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 2-fluoro-5-pyridineboronic acid was replaced with 4-fluorophenylboronic acid, and 4-bromo-2-(methylamino)pyridine was replaced with 2-pyridine. The obtained Compound I-24 was a yellow solid with a yield of 2%. ESI-MS (positive): 466.9 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 10.63 (s, 1H), 8.10 (dd, J = 5.6, 1.8 Hz, 1H), 7.90 (d, J = 8.6 Hz, 2H), 7.86–7.76 (m, 2H), 7.25–7.17 (m, 1H), 7.16–7.07 (m, 3H), 6.85 (t, J = 6.4 Hz, 1H), 3.77 (t, J = 5.0 Hz, 4H), 3.53 (t, J = 5.2 Hz, 4H).

[0479] Example I-25: Preparation of Compound I-25

[0480]

[0481] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 4-bromophenylboronic acid. The obtained Compound I-25 was a yellow solid with a yield of 19%. ESI-MS (positive): 466.8 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 9.80 (s, 1H), 8.73 (d, J = 2.5 Hz, 1H), 8.11 (dd, J = 9.0, 2.5 Hz, 1H), 7.78 (d, J = 8.7 Hz, 1H), 7.47–7.30 (m, 3H), 7.04 (d, J = 9.1 Hz, 1H), 7.01–6.90 (m, 3H), 3.79 (t, J = 5.3 Hz, 4H), 3.28 (t, J = 5.1 Hz, 4H).

[0482] Example I-26: Preparation of Compound I-26

[0483]

[0484] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 4-bromopyridine. Compound I-26 was obtained as a yellow solid with a yield of 22%. ESI-MS (positive): 489.9 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 13.42 (s, 1H), 9.82 (s, 1H), 8.75 (d, J = 2.5 Hz, 1H), 8.29 (d, J = 7.0 Hz, 2H), 8.14 (dd, J = 9.0, 2.5 Hz, 1H), 7.78 (d, J = 8.8 Hz, 1H), 7.38 (d, J = 2.4 Hz, 1H), 7.22 (d, J = 7.1 Hz, 2H), 7.08–6.89 (m, 2H), 3.88 (s, 8H).

[0485] Example I-27: Preparation of Compound I-27

[0486]

[0487] Using the synthetic method of Intermediate Ic-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 2-bromopyridine. Compound I-27 was obtained as a yellow solid with a yield of 40%. ESI-MS (positive): 403.9 [M+1] + . 11H NMR (400 MHz, Chloroform-d) δ 8.80 (s, 1H), 8.29–8.14 (m, 2H), 7.89 (d, J = 8.9 Hz, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.34 (s, 1H), 7.07 (d, J = 8.7, 2.2 Hz, 1H), 6.79–6.63 (m, 3H), 3.89 (s, 3H), 3.87–3.81 (m, 4H), 3.76–3.71 (m, 4H).

[0488] Example I-28: Preparation of Compound I-28

[0489]

[0490] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 2-bromopyridine. Compound I-28 was obtained as a yellow solid with a yield of 35%. ESI-MS (positive): 389.9 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 8.74 (d, J = 2.4 Hz, 1H), 8.16 (dd, J = 9.1, 2.4 Hz, 1H), 8.07 (d, J = 6.1 Hz, 1H), 8.02 (t, J = 8.3 Hz, 1H), 7.79 (d, J = 8.8 Hz, 1H), 7.42–7.37 (m, 2H), 7.06 (d, J = 9.1 Hz, 1H), 7.00–6.92 (m, 2H), 3.93–3.81 (m, 8H).

[0491] Example I-29: Preparation of Compound I-29

[0492]

[0493] Using the synthetic method of Intermediate Ic-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 3-bromopyridine. Compound I-29 was obtained as a yellow solid with a yield of 41%. ESI-MS (positive): 403.9 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 8.78 (d, J = 2.4 Hz, 1H), 8.49 (d, J = 2.8 Hz, 1H), 8.22–8.12 (m, 2H), 7.99 (dd, J = 8.9, 2.8 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.76 (dd, J = 8.9, 5.1 Hz, 1H), 7.69 (d, J = 2.5 Hz, 1H), 7.19–6.90 (m, 2H), 3.90–3.81 (m, 7H), 3.54 (t, 4H).

[0494] Example I-30: Preparation of Compound I-30

[0495]

[0496] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 3-bromopyridine. The obtained Compound I-30 was a yellow solid with a yield of 40%. ESI-MS (positive): 389.9 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 9.84 (s, 1H), 8.75 (d, J = 2.5 Hz, 1H), 8.49 (d, J = 2.8 Hz, 1H), 8.21 (d, J = 5.2 Hz, 1H), 8.13 (dd, J = 9.0, 2.5 Hz, 1H), 8.04 (dd, J = 9.2, 2.7 Hz, 1H), 7.83–7.75 (m, 2H), 7.38 (d, J = 2.4 Hz, 1H), 7.07 (d, J = 9.1 Hz, 1H), 6.96 (dd, J = 8.8, 2.4 Hz, 1H), 3.93–3.75 (m, 4H), 3.64–3.53 (m, 4H).

[0497] Example I-31: Preparation of Compound I-31

[0498]

[0499] Using the synthetic method of Intermediate Ic-1 in Example I-1, except that 6-methoxy-2-methylbenzothiazole was replaced with 6-dimethylamino-2-methylbenzothiazole, and 4-bromo-2-(methylamino)pyridine was replaced with 2-bromopyridine. The obtained Compound I-31 was a yellow solid with a yield of 37%. ESI-MS (positive): 417.0 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 8.73 (d, J = 2.5 Hz, 1H), 8.15–8.04 (m, 2H), 7.86 (t, J = 8.1 Hz, 1H), 7.78 (d, J = 9.0 Hz, 1H), 7.33 (d, J = 2.6 Hz, 1H), 7.22 (d, J = 9.0 Hz, 1H), 7.05–6.95 (m, 2H), 6.86 (t, J = 6.5 Hz, 1H), 3.87–3.81 (m, 4H), 3.79–3.74 (m, 4H), 2.99 (s, 6H).

[0500] Example I-32: Preparation of Compound I-32

[0501]

[0502] The preparation route of Compound I-32 is as follows:

[0503]

[0504] Preparation of Intermediate Ic-32:

[0505] The preparation of Intermediate Ic-32 adopts the synthesis method of Intermediate Ic-1 in Example I-1, except that 6-methoxy-2-bromobenzothiazole is replaced by 6-amino-2-bromobenzothiazole, and 4-bromo-2-(methylamino)pyridine is replaced by 3-bromopyridine. The obtained Compound Ic-32 is a yellow solid with a yield of 31%. ESI-MS (positive): 389.1 [M+1] + .

[0506] Preparation of Compound I-32:

[0507] Dissolve Intermediate Ic-32 (73 mg, 0.2 mmol) in 2 mL of NMP, add Cs2CO3 (195 mg, 0.6 mmol), 1-bromo-2-fluoroethane (35 μL, 0.4 mmol), and react at 100 °C for 12 h. After the reaction is completed, pour the reaction solution into 5 mL of water, extract with 20 mL of ethyl acetate, wash three times with brine, dry over anhydrous sodium sulfate, and purify by column chromatography to obtain Compound I-32 as a yellow solid with a yield of 1.8%. ESI-MS (positive): 435.0 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 8.88–8.67 (m, 1H), 8.21–8.05 (m, 2H), 7.87–7.60 (m, 2H), 7.29–6.74 (m, 6H), 4.81–4.46 (m, 2H), 4.01–3.66 (m, 8H), 3.60–3.31 (m, 2H).

[0508] Example I-33: Preparation of Compound I-33

[0509]

[0510] Using the synthetic method of Example I-32, just replace 1-bromo-2-fluoroethane with epifluorohydrin, to obtain Compound I-33 as a yellow solid, with a yield of 5%. ESI-MS (positive): 465.1 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 8.71 (s, 1H), 8.10 (d, J = 6.8 Hz, 2H), 7.86 (t, J = 8.1 Hz, 1H), 7.68 (d, J = 8.8 Hz, 1H), 7.22 (d, J = 9.2 Hz, 1H), 7.14 (s, 1H), 7.01 (d, J = 8.9 Hz, 1H), 6.91–6.80 (m, 2H), 4.55–4.44 (m, 1H), 4.43–4.30 (m, 1H), 3.97–3.92 (m, 1H), 3.86–3.83 (m, 4H), 3.79–3.75 (m, 4H), 3.21 (dd, J = 13.3, 5.8 Hz, 1H), 3.09 (dd, J = 13.2, 6.3 Hz, 1H).

[0511] Example I-34: Preparation of Compound I-34

[0512]

[0513] Using the synthetic method of Intermediate Ic-1 in Example I-1, just replace 6-methoxy-2-bromobenzothiazole with 6-dimethylamino-2-bromobenzothiazole, and replace 4-bromo-2-(methylamino)pyridine with 2-bromo-5-methoxypyridine. To obtain Compound I-34 as a yellow solid, with a yield of 9%. ESI-MS (positive): 447.1 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 8.72 (s, 1H), 8.14–8.07 (m, 1H), 7.87 (s, 1H), 7.78 (d, J = 8.9 Hz, 1H), 7.44–7.38 (m, 1H), 7.33 (s, 1H), 7.04–6.96 (m, 3H), 3.76 (s, 3H), 3.59–3.53 (m, 4H), 3.08–3.01 (m, 4H), 2.99 (s, 6H).

[0514] Example I-35: Preparation of Compound I-35

[0515]

[0516] Using the synthesis method of Example I-1, except that epifluorohydrin was replaced with 1-bromo-2-fluoroethane. Compound I-35 was obtained as a yellow solid with a yield of 12%, ESI-MS (positive): 465.2 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 8.78 (d, J = 2.5 Hz, 1H), 8.15 (dd, J = 8.9, 2.5 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.75–7.65 (m, 2H), 7.27–7.10 (m, 2H), 7.00 (d, J = 9.1 Hz, 1H), 6.50 (dd, J = 7.1, 2.4 Hz, 1H), 5.90 (d, J = 2.4 Hz, 1H), 4.87–4.82 (m, 1H), 4.75–4.71 (m, 1H), 4.37 (t, J = 3.8 Hz, 1H), 4.31–4.27 (m, 1H), 3.87–3.77 (m, 4H), 3.70–3.60 (m, 4H), 2.83 (d, J = 4.8 Hz, 3H).

[0517] Example I-36: Preparation of Compound I-36

[0518]

[0519] Using the synthesis method of Intermediate Ic-1 in Example I-1, except that 6-methoxy-2-bromobenzothiazole was replaced with 6-fluoro-2-methylbenzothiazole. Compound I-36 was obtained as a yellow solid with a yield of 20%. ESI-MS (positive): 421.3 [M+1] + . 1HNMR(400MHz, DMSO-d6) δ 12.07 (s, 1H), 8.82 (d, J = 2.5 Hz, 1H), 8.19 (dd, J = 9.0, 2.6 Hz, 1H), 8.10–7.92 (m, 2H), 7.72–7.64 (m, 1H), 7.58 (d, J = 5.3 Hz, 1H), 7.44–7.32 (m, 1H), 7.00 (d, J = 9.0 Hz, 1H), 6.61 (dd, J = 7.5, 2.5 Hz, 1H), 5.92 (s, 1H), 3.89–3.84 (m, 4H), 3.77–3.71 (m, 4H), 2.86 (d, J = 4.8 Hz, 3H).

[0520] Example I-37: Preparation of Compound I-37

[0521]

[0522] Using the synthetic method of Intermediate Ic-1 in Example I-1, but replacing 6-methoxy-2-bromobenzothiazole with 6-dimethylamino-2-bromobenzothiazole and 4-bromo-2-(methylamino)pyridine with 4-bromo-2-fluoropyridine. Compound I-37 was obtained as a yellow solid with a yield of 34%. ESI-MS (positive): 435.2 [M+1] + . 1 H NMR(400MHz, DMSO-d6) δ 8.73 (s, 1H), 8.12 (d, J = 8.9 Hz, 1H), 7.91–7.74 (m, 2H), 7.36 (s, 1H), 7.07–6.95 (m, 2H), 6.95–6.79 (m, 1H), 6.53 (s, 1H), 3.82–3.78 (m, 4H), 3.31–3.29 (m, 4H), 3.00 (s, 6H).

[0523] Example I-38: Preparation of Compound I-38

[0524]

[0525] Using the synthetic method of Example I-1, but replacing 4-bromo-2-(methylamino)pyridine with 5-bromopyrimidine. Compound I-38 was obtained as a yellow solid with a yield of 16%, ESI-MS (positive): 467.3 [M+1] + . 11H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.59 (s, 2H), 8.48 (d, J = 2.9 Hz, 1H), 8.10 (d, J = 8.8 Hz, 1H), 7.89 (d, J = 8.9 Hz, 1H), 7.70 (d, J = 2.6 Hz, 1H), 7.57 (dd, J = 8.9, 2.9 Hz, 1H), 7.12 (dd, J = 8.9, 2.6 Hz, 1H), 4.63–4.52 (m, 1H), 4.51–4.41 (m, 1H), 4.13–4.01 (m, 3H), 3.62–3.53 (m, 4H), 3.49–3.44 (m, 4H).

[0526] Example I-39: Preparation of Compound I-39

[0527]

[0528] Using the synthesis method of Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 4-bromo-N 2 ,N 6 -dimethylpyridine-2,6-diamine. Compound I-39 was obtained as a yellow solid with a yield of 10%, ESI-MS (positive): 524.3 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 10.97 (s, 1H), 8.77 (d, J = 2.5 Hz, 1H), 8.16 (dd, J = 9.0, 2.6 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.71 (d, J = 2.5 Hz, 1H), 7.12 (dd, J = 8.9, 2.6 Hz, 1H), 7.03–6.86 (m, 3H), 5.34 (s, 2H), 4.64–4.52 (m, 1H), 4.51–4.41 (m, 1H), 4.15–3.98 (m, 3H), 3.86–3.79 (m, 4H), 3.71–3.67 (m, 4H), 2.80 (d, J = 4.5 Hz, 6H).

[0529] Example I-40: Preparation of Compound I-40

[0530]

[0531] Using the synthesis method of Example I-32, except that 1-bromo-2-fluoroethane was replaced with epifluorohydrin and 4-bromo-2-(methylamino)pyridine was replaced with 2-bromopyridine. Compound I-40 was obtained as a yellow solid with a yield of 7%. ESI-MS (positive): 494.2 [M+1]+ . 1 H NMR(400 MHz, DMSO-d6) δ 12.17–11.96 (m, 1H), 8.79–8.66 (m, 1H), 8.14–8.07 (m, 1H), 7.72–7.62 (m, 2H), 7.61–7.54 (m, 1H), 7.15–7.10 (m, 1H), 7.02–6.94 (m, 1H), 6.86 (dd, J = 8.9, 2.3 Hz, 1H), 6.62 (dd, J = 7.7, 2.4 Hz, 1H), 5.92 (s, 1H), 4.56–4.43 (m, 1H), 4.43–4.31 (m, 1H), 3.97–3.79 (m, 3H), 2.86 (d, J = 4.7 Hz, 3H).

[0532] Example I-41: Preparation of Compound I-41

[0533]

[0534] Using the synthesis method of Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 5-bromo-2-(pyrrolidin-1-yl)pyridine. Compound I-41 was obtained as a yellow solid with a yield of 35%, ESI-MS (positive): 535.2 [M+1] + . 1 HNMR(400 MHz, DMSO-d6) δ 8.76 (d, J = 2.4 Hz, 1H), 8.41 (s, 1H), 8.13 (dd, J = 8.9, 2.3 Hz, 1H), 8.00 (dd, J = 9.7, 2.5 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.73 (d, J = 2.5 Hz, 1H), 7.25 (d, J = 2.6 Hz, 1H), 7.14 - 6.97 (m, 3H), 4.62–4.53 (m, 1H), 4.50 - 4.40 (m, 1H), 4.13 - 4.00 (m, 3H), 3.81 (t, J = 4.9 Hz, 4H), 3.12 (t, J = 4.9 Hz, 4H), 2.07–1.99 (m, 1H).

[0535] Example I-42: Preparation of Compound I-42

[0536]

[0537] Using the synthesis method of Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 5-bromo-6-fluoro-2-(methylamino)pyridine. The obtained compound I-42 was a yellow solid with a yield of 24%, ESI-MS(positive): 513.2[M+1] + 。 1 H NMR(400MHz,DMSO-d6)δ8.79(d,J=2.5Hz,1H),8.43(s,1H),8.12(dd,J=9.0,2.5Hz,1H),8.00(d,J=9.8Hz,1H),7.86(d,J=8.9Hz,1H),7.73(d,J=2.6Hz,1H),7.12-6.97(m,3H),4.61–4.52(m,1H),4.50-4.40(m,1H),4.15-4.00(m,3H),3.81(t,J=4.9Hz,4H),3.12(t,J=4.9Hz,4H),2.94(s,3H).

[0538] Example I-43: Preparation of Compound I-43

[0539]

[0540] Using the synthesis method of intermediate Id-1 in Example I-1, except that 4-bromo-2-(methylamino)pyridine was replaced with 5-bromo-2-(methylamino)pyridine, and 2-fluoro-5-pyridineboronic acid was replaced with (2,6-difluoropyridin-3-yl)boronic acid. The obtained compound I-43 was a yellow solid with a yield of 27%, ESI-MS(positive): 437.1[M+1] + 。 1 H NMR(400MHz,DMSO-d6)δ9.83(s,1H),8.41(s,1H),8.14(d,J=9.0Hz,1H),8.01(dd,J=9.8,2.7Hz,1H),7.85(d,J=8.9Hz,1H),7.70(d,J=2.6Hz,1H),7.25(d,J=2.7Hz,1H),7.17-7.03(m,3H),4.61–4.52(m,1H),4.51-4.45(m,1H),4.13-4.01(m,3H),3.81(t,J=5.1Hz,4H),3.23(t,J=5.1Hz,4H),2.97(s,3H).

[0541] Example I-44: Preparation of Compound I-44

[0542]

[0543] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 6-methoxy-2-bromobenzothiazole is replaced with 6-methoxy-2-bromobenzoxazole, and 4-bromo-2-(methylamino)pyridine is replaced with 5-bromo-2-(methylamino)pyridine. Compound I-44 was obtained as a yellow solid with a yield of 22%, ESI-MS (positive): 403.2 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 9.93 (s, 1H), 8.41 (s, 1H), 8.13 - 8.00 (m, 2H), 7.87 - 7.80 (m, 1H), 7.72 (d, J = 2.6 Hz, 1H), 7.23 (d, J = 2.6 Hz, 1H), 7.15 - 7.05 (m, 3H), 3.82 (t, J = 5.1 Hz, 4H), 3.21 (t, J = 5.1 Hz, 4H), 2.93 (s, 3H).

[0544] Example I-45: Preparation of Compound I-45

[0545]

[0546] Using the synthetic method of Intermediate Id-1 in Example I-1, except that 6-methoxy-2-bromobenzothiazole is replaced with 2-bromo-6-methoxythieno[2,3-b]pyridine, and 4-bromo-2-(methylamino)pyridine is replaced with 5-bromo-2-(methylamino)pyridine. Compound I-45 was obtained as a yellow solid with a yield of 13%, ESI-MS (positive): 419.2 [M+1] + 。 1 H NMR (400 MHz, DMSO-d6) δ 10.98 (s, 1H), 8.45 (s, 1H), 7.78 - 7.66 (m, 3H), 7.17 - 7.07 (m, 2H), 6.82 - 6.54 (m, 4H), 3.81 (t, J = 5.0 Hz, 4H), 3.25 (t, J = 5.0 Hz, 4H), 2.90 (s, 3H).

[0547] Example I-46: Preparation of Compound I-46

[0548]

[0549] Synthetic route of Compound I-46:

[0550]

[0551] The first step: Preparation of Intermediate Ie-1

[0552] Glycerol (1 g, 11 mmol) and 4-dimethylaminopyridine (DMAP) (335 mg, 2.75 mmol) were dissolved in pyridine (10 mL). At 0 °C, p-toluenesulfonyl chloride (TsCl) (4.4 g, 23 mmol) dissolved in pyridine (15 mL) was added dropwise, and the reaction was carried out overnight at room temperature. After the reaction was complete, the reaction solution was diluted in 100 mL of water, and the pH was adjusted to acidic by dropping concentrated hydrochloric acid. It was extracted with DCM (100 mL × 5), and the organic phase was washed with 1N HCl (100 mL), then washed with water (100 mL). Finally, it was purified by flash silica gel column chromatography (PE / EA) to obtain product Ie-1 (2.8 g, 7 mmol), a colorless oily liquid, with a yield of 64%. MS-ESI: m / z 401 [M+H] + 。

[0553] Step 2: Preparation of intermediate If-1

[0554] Ie-1 (1.2 g, 3 mmol) and 3,4-dihydropyran (DHP) (5.5 mL, 60 mmol) were dissolved in DCM (10 mL), and pyridinium p-toluenesulfonate (PPTS) (150 mg, 0.6 mmol) was added. The reaction was carried out overnight at 40 °C. After the reaction was complete, the reaction solution was concentrated and purified by flash silica gel column chromatography (PE / EA) to obtain product If-1 (1.3 g, 2.7 mmol), a pale yellow oily liquid, with a yield of 89%. MS-ESI: m / z 485 [M+H] + 。

[0555] Step 3: Preparation of product I-46

[0556] Id-2 (0.06 mmol), If-1 (0.29 mmol) and potassium carbonate (16 mg, 0.12 mmol) were added to DMF (0.5 mL), and the reaction was carried out at 80 °C for 2 h. After the reaction was complete, it was purified by flash silica gel column chromatography (PE / EA) to obtain product I-46 (3.8 mg, 0.006 mmol), a yellow solid, with a yield of 69%. MS-ESI: m / z 731.2 [M+H] + 。

[0557] Example I-47: Preparation of compound I-47

[0558]

[0559] Method 1:

[0560]

[0561] 150 mCi 18F is enriched on the QMA column and eluted with 0.6 mL of eluent (150 mg of Kryptofix 222 and 15 mg of K2CO3 dissolved in 9 mL of acetonitrile and 1 mL of water) into the reaction flask. 18 The F is eluted to the reaction flask; under high-purity nitrogen, the solvent is dried by heating at 110 °C for 20 min; 0.2 mg of 2-(oxiran-2-ylmethyl)-4-methylbenzenesulfonate and 0.2 mL of acetonitrile are added to the reaction flask, and the reaction is carried out at 90 °C for 10 min; the acetonitrile is evaporated to dryness, 0.2 mL of DMF solvent and 0.5 mg of Id-2 are added, and the reaction is carried out at 130 °C for 10 min; the mixture is separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 26:74, flow rate: 3 mL / min), the product is collected and solid-phase extracted with a C18 column, and the final product I-47 is obtained by eluting with ethanol, and the labeling efficiency is 8%.

[0562] Method 2:

[0563]

[0564] 150 mCi 18 F is enriched on the QMA column and eluted with 0.6 mL of eluent (150 mg of Kryptofix 222 and 15 mg of K2CO3 dissolved in 9 mL of acetonitrile and 1 mL of water) into the reaction flask. 18 The F is eluted to the reaction flask; under high-purity nitrogen, the solvent is dried by heating at 110 °C for 20 min; 0.5 mg of I-46 is added to the reaction flask, and the reaction is carried out at 140 °C for 10 min; then 0.2 mL of 2N hydrochloric acid is added and the reaction is carried out at 105 ℃ for 8 min to remove the THP protecting group; the reaction solution is separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 26:74, flow rate: 3 mL / min), the product is collected and solid-phase extracted with a C18 column, and the final product I-47 is obtained by eluting with ethanol, and the labeling efficiency is 10%.

[0565] Example I-48: Preparation of Compound I-48

[0566]

[0567] The synthesis method of Example I-47 is adopted, except that the intermediate is replaced with the corresponding raw material.

[0568] Example I-49: Preparation of Compound I-49

[0569]

[0570] Synthesis route of Compound I-49:

[0571]

[0572] Step 1: Synthesis of Precursor I-49pre

[0573] Id-1 (0.06 mmol), diethylene glycol xylene sulfonate (0.29 mmol) and potassium carbonate (16 mg, 0.12 mmol) were added to DMF (0.5 mL), and the reaction was carried out at 80 °C for 2 h. After the reaction was complete, it was purified by flash silica gel column chromatography (PE / EA) to obtain the product I-49pre (3.8 mg, 0.006 mmol), a yellow solid, with a yield of 83%. MS-ESI: m / z 617.2 [M+H] + 。

[0574] Step 2: Radioactive Labeling of I-49

[0575] 150 mCi 18 F was enriched on a QMA column and eluted with 0.6 mL of eluent (150 mg of Kryptofix 222, 15 mg of K2CO3 dissolved in 9 mL of acetonitrile and 1 mL of water) to 18 F was eluted into the reaction flask; under high-purity nitrogen, the solvent was dried by heating at 110 °C for 20 min; 0.5 mg of I-49pre was added to the reaction flask and reacted at 140 °C for 10 min; the reaction solution was separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 29:71, flow rate: 3 mL / min), the product was collected and solid-phase extracted with a C18 column, and eluted with ethanol to obtain the final product I-49, and the labeling efficiency was 18%.

[0576] Example I-50: Preparation of Compound I-50

[0577]

[0578] Synthesis Route of Compound I-50:

[0579]

[0580] Step 1: Synthesis of Precursor I-50pre

[0581] The synthesis method was the same as that of Example I-43, except that 2-fluoro-5-boronic acid pyridine was replaced by 2-fluoro-5-boronic acid-6-fluoro-pyridine. MS-ESI: m / z 464.1 [M+H] + 。

[0582] Step 2: Radioactive Labeling of I-50

[0583] Enrich 150 mCi 18 F on a QMA column and elute 18 F with 0.6 mL of eluent (dissolved 150 mg of Kryptofix 222 and 15 mg of K2CO3 in 9 mL of acetonitrile and 1 mL of water) into the reaction flask; under high-purity nitrogen, heat at 110 °C for 20 min to dry the solvent; add 0.7 mg of I-50pre to the reaction flask, ℃ react for 10 min; separate and purify the reaction solution by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 20:80, flow rate: 3 mL / min), collect the product and perform solid-phase extraction with a C18 column, elute with ethanol to obtain the final product I-50, and the labeling efficiency is 5%.

[0584] Example I-51: Preparation of Compound I-51

[0585]

[0586] Adopt the synthesis method of Example I-1, just replace 4-bromo-2-(methylamino)pyridine with 4-bromo-2-aminopyridine. Compound I-51 is obtained as a yellow solid with a yield of 12%, ESI-MS (positive): 481.2 [M+1] + . 1 1H NMR (400 MHz, DMSO-d6) δ 12.08 (s, 1H), 8.77 (s, 1H), 8.16 (d, J = 9.0 Hz, 1H), 7.87 (d, J = 8.9 Hz, 1H), 7.77–7.62 (m, 2H), 7.42–6.91 (m, 4H), 6.77–6.57 (m, 1H), 6.17–5.94 (m, 1H), 4.66–4.51 (m, 1H), 4.51–4.37 (m, 1H), 4.15–4.01 (m, 3H), 3.87–3.83 (m, 4H), 3.69–3.66 (m, 4H).

[0587] Example II-1: Preparation of Compound II-1

[0588]

[0589] Synthesis route of Compound II-1:

[0590]

[0591] First step: Preparation of intermediate IIa-1

[0592] (6-Formylpyridin-3-yl)boronic acid (48 mg, 0.32 mmol), 5-bromo-2-(methylamino)pyridine (60 mg, 0.32 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (Pd(dppf)Cl2) (23 mg, 0.032 mmol) and potassium carbonate (138 mg, 0.96 mmol) were dissolved in 1,4-dioxane (2.4 mL) and water (0.6 mL). After nitrogen protection, the reaction was carried out overnight at 80 °C. After the reaction was complete, it was purified by flash silica gel column chromatography, petroleum ether (PE) / ethyl acetate (EA), to obtain product IIa-1 (42 mg, 0.2 mmol), a yellow solid, with a yield of 61%. MS-ESI: m / z 214 [M+H] + .

[0593] Step 2: Preparation of intermediate IIb-1

[0594] IIa-1 (42 mg, 0.2 mmol) and 6-methoxy-2-methylbenzothiazole (36 mg, 0.2 mmol) were dissolved in dimethyl sulfoxide (DMSO) (2 mL), and an aqueous sodium hydroxide solution (1 g / mL) (32 μL, 0.8 mmol) was added. The reaction was carried out at 50 °C for 2 h. After the reaction was complete, it was purified by flash silica gel column chromatography (PE / EA) and flash C18 column chromatography (acetonitrile / water) to obtain product IIb-1 (4 mg, 0.01 mmol), an orange solid, with a yield of 5%. MS-ESI: m / z 375 [M+H] + 。

[0595] Step 3: Preparation of intermediate IIc-1

[0596] IIb-1 (4 mg, 0.01 mmol) was dissolved in dichloromethane (DCM) (0.03 mL). Under nitrogen protection, a dichloromethane solution of boron tribromide (1 M) (0.03 mL, 0.03 mmol) was added dropwise at -78 °C. After stirring for 2 h, it was restored to room temperature and the reaction was carried out overnight. The reaction solution was quenched by dropping it into a saturated aqueous sodium bicarbonate solution (50 mL) and extracted with EA (50 mL). The EA phase was washed with water (50 mL × 2) and then washed with a saturated aqueous sodium chloride solution (50 mL), and dried over anhydrous sodium sulfate to obtain product IIc-1 (3.6 mg, 0.01 mmol), an orange solid, with a yield of 93%. MS-ESI: m / z 361 [M+H] + 。 11H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 8.57 (d, J = 2.4 Hz, 1H), 8.37 (d, J = 4.8, 1.9 Hz, 1H), 8.09 (dd, J = 9.0, 2.4 Hz, 1H), 7.82 - 7.72 (m, 2H), 7.47 (s, 2H), 7.41 - 7.34 (m, 2H), 7.25 (d, J = 8.8 Hz, 1H), 7.08 (dd, J = 7.2, 4.9 Hz, 1H), 6.96 (dd, J = 8.8, 2.5 Hz, 1H), 3.57 (s, 3H).

[0597] Step 4: Preparation of Product II-1

[0598] Add IIc-1 (3.6 mg, 0.01 mmol) and potassium carbonate (4 mg, 0.03 mmol) to N,N-dimethylformamide (DMF) (0.1 mL), dropwise add epifluorohydrin (1.5 μL, 0.02 mmol), and react at 80 °C for 4 h. After the reaction is complete, purify using high performance liquid chromatography (HPLC) (acetonitrile / water) to obtain Product II-1 (0.9 mg, 0.002 mmol), an orange solid, with a yield of 21%. MS-ESI: m / z 437 [M+H] + 。 1 1H NMR (400 MHz, DMSO-d6) δ 8.98 (d, J = 2.4 Hz, 1H), 8.42 (s, 1H), 8.23 - 8.12 (m, 2H), 7.92 (s, 1H), 7.89 (d, J = 8.5 Hz, 1H), 7.84 (d, J = 8.2 Hz, 1H), 7.74 (d, J = 2.5 Hz, 1H), 7.65 (d, J = 15.8 Hz, 1H), 7.16 (dd, J = 8.9, 2.6 Hz, 1H), 7.00 - 6.86 (m, 1H), 4.63–4.52 (m, 1H), 4.52 - 4.41 (m, 1H), 4.17–4.02 (m, 3H), 2.94 (s, 3H).

[0599] Resolution and Characterization of Isomers of Compound II-1

[0600]

[0601] Compound II-1 (100 mg, 0.23 mmol) was dissolved in MeOH:DCM = 1:1 (80 mL) and separated and purified by chiral SFC (column: ChiralPak IH, 250×40 mm I.D., 10 μm, mobile phase: A: CO2, B: MeOH (0.1% NH3.H2O), flow rate: 140 mL / min) to obtain yellow solid II-1A (shorter retention time, 40 mg, MS-ESI: m / z 437.1 [M+H]+) and yellow solid II-1B (longer retention time, 40 mg, MS-ESI: m / z 437.0 [M+H]+).

[0602] Single configuration compound I-1A (shorter retention time):

[0603] SFC analysis: retention time: 4.578 min, ee value > 99.9% (column: Chiral Pak IH, 100×3 mm I.D., 3 μm, mobile phase: A: CO2, B: MeOH (0.1% DEA), flow rate: 2.0 mL / min).

[0604] 1 HNMR (400 MHz, DMSO-d6) δ 8.93 (d, J = 2.3 Hz, 1H), 8.48 (d, J = 2.5 Hz, 1H), 8.06 (dd, J = 8.2, 2.4 Hz, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.89–7.80 (m, 2H), 7.76 (d, J = 8.2 Hz, 1H), 7.73 (d, J = 2.6 Hz, 1H), 7.61 (d, J = 15.9 Hz, 1H), 6.57 (d, J = 8.8 Hz, 1H), 5.53 (d, J = 5.1 Hz, 1H), 4.58 (qd, J = 9.6, 4.2 Hz, 1H), 4.52–4.40 (m, 1H), 4.18–3.96 (m, 3H), 2.83 (d, J = 4.8 Hz, 3H).

[0605] Single configuration compound I-1B (longer retention time):

[0606] SFC analysis: retention time: 5.744 min, ee value: 99.59% (column: Chiral Pak IH, 100×3 mm I.D., 3 μm, mobile phase: A: CO2, B: MeOH (0.1% DEA), flow rate: 2.0 mL / min).

[0607] 1HNMR(400MHz, DMSO-d6) δ 8.93 (d, J = 2.3 Hz, 1H), 8.48 (d, J = 2.5 Hz, 1H), 8.06 (dd, J = 8.2, 2.4 Hz, 1H), 7.90 (d, J = 8.9 Hz, 1H), 7.89–7.80 (m, 2H), 7.76 (d, J = 8.2 Hz, 1H), 7.73 (d, J = 2.6 Hz, 1H), 7.61 (d, J = 15.9 Hz, 1H), 6.57 (d, J = 8.8 Hz, 1H), 5.53 (d, J = 5.1 Hz, 1H), 4.58 (qd, J = 9.6, 4.2 Hz, 1H), 4.52–4.40 (m, 1H), 4.18–3.96 (m, 3H), 2.83 (d, J = 4.8 Hz, 3H).

[0608] Example II-2: Preparation of Compound II-2

[0609]

[0610] Example II-2 is intermediate IIc-1, and its synthesis details can be seen in Example II-1.

[0611] Example II-3: Preparation of Compound II-3

[0612]

[0613] The synthesis method of Example 3 is the same as that of intermediate IIc-1, except that (6-formylpyridin-3-yl)boronic acid is replaced by 4-borobenzaldehyde, and 5-bromo-2-(methylamino)pyridine is replaced by 3-bromopyridine. The obtained product II-3 is a yellow solid with a yield of 26%. LC-MS(ESI)[M + H] + : 331.0. 1 H NMR(400MHz, DMSO-d6) δ 9.91 (s, 1H), 9.09 (d, J = 2.4 Hz, 1H), 8.73 - 8.66 (m, 1H), 8.41 (d, J = 8.0 Hz, 1H), 7.98 - 7.83 (m, 4H), 7.79 (d, J = 8.8 Hz, 1H), 7.72 (dd, J = 8.0, 5.0 Hz, 1H), 7.65 (d, J = 16.2 Hz, 1H), 7.57 (d, J = 16.3 Hz, 1H), 7.38 (d, J = 2.5 Hz, 1H), 6.98 (dd, J = 8.8, 2.5 Hz, 1H).

[0614] Example II-4: Preparation of Compound II-4

[0615]

[0616] The synthesis method of Example II-4 is the same as that of Intermediate IIc-1, except that (6-formylpyridin-3-yl)boronic acid is replaced with 4-borobenzaldehyde. The obtained product II-4 is a yellow solid with a yield of 31%. LC-MS (ESI) [M+H] + : 360.1. 1 H NMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 8.56–8.11 (m, 3H), 7.84 (s, 2H), 7.80–7.69 (m, 3H), 7.57 (dd, J = 16.1, 4.2 Hz, 2H), 7.37 (s, 1H), 7.08–6.88 (m, 2H), 2.96 (d, J = 4.1 Hz, 3H).

[0617] Example II-5: Preparation of Compound II-5

[0618]

[0619] The synthesis method of Example II-5 is the same as that of Example II-1, except that (6-formylpyridin-3-yl)boronic acid is replaced with 4-borobenzaldehyde and 5-bromo-2-(methylamino)pyridine is replaced with 4-bromo-2-(methylamino)pyridine. The obtained product II-5 is a yellow solid with a yield of 12%. LC-MS (ESI) [M+H] + : 436.0. 1 H NMR (400 MHz, DMSO-d6) δ 8.03 (d, J = 6.6 Hz, 1H), 7.97 (d, J = 8.2 Hz, 2H), 7.94–7.86 (m, 3H), 7.77–7.71 (m, 2H), 7.65 (d, J = 16.2 Hz, 1H), 7.26 (d, J = 8.3 Hz, 2H), 7.15 (dd, J = 8.9, 2.6 Hz, 1H), 4.64–4.53 (m, 1H), 4.51–4.39 (m, 1H), 4.16–4.02 (m, 3H), 3.00 (s, 3H).

[0620] Example II-6: Preparation of Compound II-6

[0621]

[0622] The synthesis method of Example II-6 is the same as that of Example II-1, except that (6-formylpyridin-3-yl)boronic acid is replaced with 3-borobenzaldehyde and 5-bromo-2-(methylamino)pyridine is replaced with 4-bromo-2-(methylamino)pyridine. The obtained product II-6 is a yellow solid with a yield of 12%. LC-MS (ESI) [M+H]+ : 436.0 1 H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.07 (d, J = 6.8 Hz, 1H), 7.96 (d, J = 7.3 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.85–7.78 (m, 2H), 7.77–7.69 (m, 2H), 7.67–7.62 (m, 1H), 7.55–7.46 (m, 1H), 7.34–7.29 (m, 1H), 7.15 (dd, J = 8.9, 2.5 Hz, 1H), 7.10–7.00 (m, 1H), 4.62–4.50 (m, 1H), 4.48–4.37 (m, 1H), 4.14–3.92 (m, 3H), 3.02 (s, 3H).

[0623] Example II-7: Preparation of Compound II-7

[0624]

[0625] Synthetic route of Compound II-7:

[0626]

[0627] The first step: Preparation of intermediate IId-1

[0628] Glycerol (1 g, 11 mmol) and 4-dimethylaminopyridine (DMAP) (335 mg, 2.75 mmol) were dissolved in pyridine (10 mL). p-Toluenesulfonyl chloride (TsCl) (4.4 g, 23 mmol) dissolved in pyridine (15 mL) was added dropwise at 0 °C, and the reaction was carried out overnight at room temperature. After the reaction was complete, the reaction solution was diluted in 100 mL of water, and the pH was adjusted to acidic by dropping concentrated hydrochloric acid. It was extracted with DCM (100 mL × 5), the organic phase was washed with 1N HCl (100 mL), then washed with water (100 mL), and finally purified by flash silica gel column chromatography (PE / EA) to obtain the product IId-1 (2.8 g, 7 mmol), a colorless oily liquid, with a yield of 64%. MS-ESI: m / z 401 [M+H] + .

[0629] The second step: Preparation of intermediate IIe-1

[0630] IId-1 (1.2 g, 3 mmol) and 3,4-dihydropyran (DHP) (5.5 mL, 60 mmol) were dissolved in DCM (10 mL). Pyridinium p-toluenesulfonate (PPTS) (150 mg, 0.6 mmol) was added and allowed to react at 40°C overnight. After the reaction was complete, the reaction solution was concentrated and purified by flash silica gel column chromatography (PE / EA) to obtain the product IIe-1 (1.3 g, 2.7 mmol) as a light yellow oil in 89% yield. MS-ESI: m / z 485 [M+H] + .

[0631] Step 3: Preparation of product II-7

[0632] IIc-1 (21 mg, 0.06 mmol), IIe-1 (140 mg, 0.29 mmol), and potassium carbonate (16 mg, 0.12 mmol) were added to DMF (0.5 mL) and reacted at 80°C for 2 h. After the reaction was complete, it was purified by flash silica gel column chromatography (PE / EA) to obtain the product II-7 (3.8 mg, 0.006 mmol) as a yellow solid in a 10% yield. MS-ESI: m / z 673 [M+H] + .

[0633] Example II-8: Preparation of Compound II-8

[0634]

[0635] Method 1:

[0636]

[0637] 150mCi 18 F was enriched on a QMA column and washed with 0.6 mL of eluent (150 mg 222, 15mg K2CO3 dissolved in 9mL acetonitrile and 1mL water) 18 F was eluted into a reaction flask; the solvent was dried by heating at 110°C for 20 min under high-purity nitrogen; 0.2 mg of oxirane-2-ylmethyl 4-methylbenzenesulfonate and 0.2 mL of acetonitrile were added to the reaction flask, and the mixture was reacted at 90°C for 10 min; the acetonitrile was evaporated to dryness, and 0.2 mL of DMF solvent and 0.5 mg of IIc-1 were added, and the mixture was reacted at 130°C for 10 min; the mixture was separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile:water = 26:74, flow rate: 3 mL / min, peak elution time: 25 min), the product was collected and subjected to solid-phase extraction on a C18 column, and eluted with ethanol to obtain the final product II-8, with a labeling efficiency of 15%.

[0638] Method 2:

[0639]

[0640] Enrich 150 mCi 18 F on a QMA column and elute 18 F with 0.6 mL of eluent (dissolving 150 mg of Kryptofix 222 and 15 mg of K2CO3 in 9 mL of acetonitrile and 1 mL of water) into the reaction flask; under high-purity nitrogen, heat at 110 °C for 20 min to dry the solvent; add 0.5 mg of II-7 to the reaction flask and react at 140 °C for 10 min; then add 0.2 mL of 2N hydrochloric acid and react at 105 °C for 8 min to remove the THP protecting group; separate and purify the reaction solution by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 26:74, flow rate: 3 mL / min, peak elution time: 25 min), collect the product and perform solid-phase extraction with a C18 column, and elute with ethanol to obtain the final product II-8, with a labeling efficiency of 13%.

[0641] According to a method similar to that of Example II-1, for example, using a ChiraPak IH column, a pair of enantiomers of Example II-8 can be resolved: enantiomer 1 and enantiomer 2.

[0642] Example II-9: Preparation of Compound II-9

[0643]

[0644] The synthesis method of Example II-9 is the same as that of Example II-1, except that 6-methoxy-2-methylbenzothiazole is replaced by 6-methoxy-2-methylbenzoxazole. The obtained product II-9 is a yellow solid with a yield of 10%. LC-MS (ESI) [[M+H]] + : 421.2. 1 H NMR (400 MHz, DMSO-d6) δ 9.01 (d, J = 2.4 Hz, 1H), 8.41 (s, 1H), 8.25–8.14 (m, 2H), 7.89 (d, J = 8.2 Hz, 1H), 7.80 (d, J = 16.0 Hz, 1H), 7.67 (d, J = 8.7 Hz, 1H), 7.59 (d, J = 15.9 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 7.09–6.95 (m, 2H), 4.65–4.52 (m, 1H), 4.51–4.39 (m, 1H), 4.15–4.01 (m, 3H), 2.96 (s, 3H).

[0645] Example II-10: Preparation of Compound II-10

[0646]

[0647] The synthesis method of Example II-10 is the same as that of Example II-1, except that 6-methoxy-2-methylbenzothiazole is replaced by 7-methoxy-2-methylbenzothiazole. The obtained product II-10 is a yellow solid with a yield of 19%. LC-MS (ESI) [M+H] + : 437.2. 1 H NMR (400 MHz, DMSO-d6) δ 9.01 (d, J = 2.4 Hz, 1H), 8.40 (s, 1H), 8.26 (d, J = 9.2 Hz, 1H), 8.19 (dd, J = 8.2, 2.5 Hz, 1H), 8.01 (d, J = 8.8 Hz, 1H), 7.93 (d, J = 15.9 Hz, 1H), 7.88 (d, J = 8.2 Hz, 1H), 7.73 (d, J = 15.9 Hz, 1H), 7.58 (d, J = 2.5 Hz, 1H), 7.14 (dd, J = 8.8, 2.5 Hz, 1H), 7.04 (d, J = 9.3 Hz, 1H), 4.64–4.53 (m, 1H), 4.52–4.40 (m, 1H), 4.16–4.04 (m, 3H), 2.97 (s, 3H).

[0648] Example II-11: Preparation of Compound II-11

[0649]

[0650] The synthesis method of Example II-11 is the same as that of Example II-1, except that (6-formylpyridin-3-yl)boronic acid is replaced by (6-formylpyridin-4-yl)boronic acid. The obtained product II-11 is a yellow solid with a yield of 17%. LC-MS (ESI) [M+H] + : 437.2. 11H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 5.4 Hz, 1H), 8.58 (s, 1H), 8.28 (d, J = 9.0 Hz, 1H), 8.24 (s, 1H), 8.04 (d, J = 16.0 Hz, 1H), 7.93 (d, J = 8.9 Hz, 1H), 7.81–7.74 (m, 2H), 7.65 (d, J = 16.0 Hz, 1H), 7.18 (dd, J = 8.9, 2.5 Hz, 1H), 6.97 (d, J = 9.1 Hz, 1H), 4.64–4.53 (m, 1H), 4.51–4.41 (m, 1H), 4.14–4.03 (m, 3H), 2.97 (s, 3H).

[0651] Example II-12: Preparation of Compound II-12

[0652]

[0653] The synthesis method of Example II-12 is the same as that of Example II-1, except that 5-bromo-2-(methylamino)pyridine is replaced by 4-bromo-2-(methylamino)pyridine. The obtained product II-12 is a yellow solid with a yield of 13%. LC-MS (ESI) [M+H] + : 437.1. 1 1H NMR (400 MHz, DMSO-d6) δ 9.11 (s, 1H), 8.36–8.28 (m, 1H), 8.08 (d, J = 6.5 Hz, 1H), 8.03–7.89 (m, 3H), 7.75 (d, J = 2.6 Hz, 1H), 7.70 (d, J = 15.8 Hz, 1H), 7.36–7.26 (m, 2H), 7.17 (dd, J = 8.9, 2.6 Hz, 1H), 4.64–4.51 (m, 1H), 4.51–4.41 (m, 1H), 4.16–4.03 (m, 3H), 3.00 (s, 3H).

[0654] Example II-13: Preparation of Compound II-13

[0655]

[0656] The synthesis method of Example II-13 is the same as that of Intermediate IIb-1, except that 6-methoxy-2-methylbenzothiazole is replaced by 6-dimethylamino-2-methylbenzothiazole. The obtained product II-13 is a yellow solid with a yield of 12%. LC-MS (ESI) [M+H] + : 388.2.

[0657] Example II-14: Preparation of Compound II-14

[0658]

[0659] The synthesis method of Example II-14 is the same as that of Example II-1, except that 5-bromo-2-(methylamino)pyridine is replaced by 4-bromo-2-(methylamino)thiazole. The obtained product II-14 is a yellow solid with a yield of 18%. LC-MS (ESI) [M+H] + : 443.1. 1 HNMR (400 MHz, DMSO-d6) δ 9.12 (d, J = 2.2 Hz, 1H), 8.25 (dd, J = 8.1, 2.3 Hz, 1H), 7.96–7.69 (m, 5H), 7.61 (d, J = 15.9 Hz, 1H), 7.37 (s, 1H), 7.16 (dd, J = 8.9, 2.6 Hz, 1H), 4.64–4.52 (m, 1H), 4.51–4.40 (m, 1H), 4.12–4.03 (m, 3H), 2.91 (s, 3H).

[0660] Example II-15: Preparation of Compound II-15

[0661]

[0662] The synthesis method of Example II-15 is the same as that of Example II-1, except that 5-bromo-2-(methylamino)pyridine is replaced by 5-bromo-2-(methylamino)thiazole. The obtained product II-15 is a yellow solid with a yield of 22%. LC-MS (ESI) [M+H] + : 443.0. 1 HNMR (400 MHz, DMSO-d6) δ 9.80 (s, 2H), 8.87 (d, J = 2.4 Hz, 1H), 8.17 (s, 1H), 8.04 (dd, J = 8.1, 2.5 Hz, 1H), 7.94–7.81 (m, 3H), 7.74 (d, J = 2.6 Hz, 1H), 7.64 (d, J = 15.8 Hz, 1H), 7.16 (dd, J = 8.9, 2.6 Hz, 1H), 4.65–4.53 (m, 1H), 4.51–4.39 (m, 1H), 4.15–4.01 (m, 3H), 3.65 (s, 3H).

[0663] Example II-16: Preparation of Compound II-16

[0664]

[0665] The synthesis method of Example II-16 is the same as that of Example II-1, except that 5-bromo-2-(methylamino)pyridine is replaced with 4-bromo-N-methylaniline. The obtained product II-16 is a yellow solid with a yield of 7%. LC-MS (ESI) [M+H] + : 436.1. 1 1H NMR (400 MHz, DMSO-d6) δ 8.92 (d, J = 2.6 Hz, 1H), 8.40 (s, 1H), 8.25 - 8.10 (m, 2H), 7.90 (s, 1H), 7.82 (d, J = 8.5 Hz, 1H), 7.80 (d, J = 8.2 Hz, 1H), 7.73 (d, J = 2.3 Hz, 2H), 7.65 (d, J = 15.6 Hz, 2H), 7.16 (dd, J = 8.9, 2.6 Hz, 1H), 7.00 - 6.86 (m, 1H), 4.63 - 4.48 (m, 2H), 4.15 - 4.03 (m, 3H), 2.96 (s, 3H).

[0666] Example II-17: Preparation of Compound II-17

[0667]

[0668] The synthesis method of Example II-17 is the same as that of Example II-14, except that the amount of epifluorohydrin used is tripled. The obtained product II-17 is a yellow solid with a yield of 4%. LC-MS (ESI) [M+H] + : 519.1. 1 1H NMR (400 MHz, DMSO-d6) δ 9.16 (s, 1H), 8.26 (s, 1H), 7.97–7.70 (m, 4H), 7.66–7.58 (m, 1H), 7.47 (d, J = 4.9 Hz, 1H), 7.17 (s, 1H), 4.62–4.30 (m, 4H), 4.20–4.01 (m, 6H), 3.20–3.13 (m, 3H).

[0669] Example II-18: Preparation of Compound II-18

[0670]

[0671] The synthesis method of Example II-18 is the same as that of Example II-1, except that (6-formylpyridin-3-yl)boronic acid is replaced with (6-formylpyrazin-3-yl)boronic acid. The obtained product II-18 is a yellow solid with a yield of 7%. LC-MS (ESI) [M+H] + : 438.2. 11H NMR (400 MHz, DMSO-d6) δ 9.30–9.26 (m, 1H), 8.97–8.90 (m, 1H), 8.80–8.74 (m, 1H), 8.41 (d, J = 9.2 Hz, 1H), 7.99–7.88 (m, 2H), 7.80–7.65 (m, 2H), 7.17 (dd, J = 8.9, 2.6 Hz, 1H), 6.91 (d, J = 9.2 Hz, 1H), 4.64–4.54 (m, 1H), 4.51–4.40 (m, 1H), 4.15–4.03 (m, 3H), 2.95 (s, 3H).

[0672] Example II-19: Preparation of Compound II-19

[0673]

[0674] The synthesis method of Example II-19 is the same as that of Example II-1, except that epichlorohydrin is replaced by 1-bromo-2-fluoroethane. The obtained product II-19 is a yellow solid with a yield of 27%. LC-MS (ESI) [[M+H]] + : 407.1. 1 1H NMR (400 MHz, DMSO-d6) δ 8.99 (s, 1H), 8.41 (s, 1H), 8.23–8.11 (m, 2H), 7.96–7.81 (m, 3H), 7.75 (s, 1H), 7.65 (d, J = 15.8 Hz, 1H), 7.18 (d, J = 9.1 Hz, 1H), 6.97 (d, J = 9.0 Hz, 1H), 4.91–4.83 (m, 1H), 4.80–4.72 (m, 1H), 4.44–4.35 (m, 1H), 4.34–4.29 (m, 1H), 2.95 (s, 3H).

[0675] Example II-20: Preparation of Compound II-20

[0676]

[0677] The synthesis method of Example II-20 is the same as that of Intermediate IIb-1, except that 6-methoxy-2-methylbenzothiazole is replaced by 6-nitro-2-methylbenzothiazole. The obtained product II-20 is a yellow solid with a yield of 12%. LC-MS (ESI) [[M+H]] + : 390.1.

[0678] Example II-21: Preparation of Compound II-21

[0679]

[0680] The synthesis method of Example II-21 is the same as that of Example II-1, except that epichlorohydrin is replaced by 1-bromo-2-oxo-p-toluenesulfonyl-ethane. The obtained product II-21 is a yellow solid with a yield of 21%. LC-MS (ESI) [M+H] + : 559.1.

[0681] Example II-22: Preparation of Compound II-22

[0682]

[0683] 150 mCi of 18 F was enriched on a QMA column, and 18 F was eluted into the reaction flask with 0.6 mL of eluent (150 mg of Kryptofix 222 and 15 mg of K2CO3 dissolved in 9 mL of acetonitrile and 1 mL of water); under high-purity nitrogen, the solvent was dried by heating at 110 °C for 20 min; 0.5 mg of the precursor compound II-21 was added to the reaction flask and reacted at 140 °C for 10 min; the reaction solution was separated and purified by HPLC), the product was collected and solid-phase extracted with a C18 column, and the final product II-22 was obtained by eluting with ethanol, and the labeling efficiency was 21%.

[0684] Example II-23: Preparation of Compound II-23

[0685]

[0686] The synthesis method of Example II-23 is the same as that of Example II-2, except that 5-bromo-2-(methylamino)pyridine is replaced by 5-bromo-6-fluoro-2-(methylamino)pyridine. The obtained product II-23 is a brown solid with a yield of 9%. LC-MS (ESI) [M+H] + : 379.1.

[0687] Example II-24: Preparation of Compound II-24

[0688]

[0689] The synthesis method of Example II-24 is the same as that of Example II-1, except that 6-methoxy-2-methylbenzothiazole is replaced by 6-amino-2-methylbenzothiazole. The obtained product II-24 is a yellow solid with a yield of 3%. LC-MS (ESI) [M+H] + : 437.2.

[0690] Example II-25: Preparation of Compound II-25

[0691]

[0692] Synthetic route of Compound II-25:

[0693]

[0694] First step: Preparation of intermediate IIa-1'

[0695] Dissolve (6-formylpyridin-3-yl)boronic acid (48 mg, 0.32 mmol), 5-bromo-2-(methylamino)pyrimidine (60 mg, 0.32 mmol), 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (Pd(dppf)Cl2) (23 mg, 0.032 mmol) and potassium carbonate (138 mg, 0.96 mmol) in 1,4-dioxane (2.4 mL) and water (0.6 mL). After nitrogen protection, react at 80 °C overnight. After the reaction is complete, purify by flash silica gel column chromatography, petroleum ether (PE) / ethyl acetate (EA), to obtain the product IIa-1' (42 mg, 0.2 mmol), a yellow solid, with a yield of 73%. MS-ESI: m / z 215.0 [M+H] + .

[0696] Second step: Preparation of intermediate IIb-1'

[0697] Dissolve IIa-1' (42 mg, 0.2 mmol) and 6-methoxy-2-methylbenzothiazole (36 mg, 0.2 mmol) in dimethyl sulfoxide (DMSO) (2 mL), add an aqueous solution of sodium hydroxide (1 g / mL) (32 μL, 0.8 mmol), and react at 50 °C for 2 h. After the reaction is complete, purify by flash silica gel column chromatography (PE / EA) and flash C18 column chromatography (acetonitrile / water) to obtain the product IIb-1' (4 mg, 0.01 mmol), an orange solid, with a yield of 11%. MS-ESI: m / z 376.1 [M+H] + 。

[0698] Third step: Preparation of intermediate IIc-1'

[0699] Dissolve IIb-1’ (4 mg, 0.01 mmol) in dichloromethane (DCM) (0.03 mL). Under nitrogen protection, dropwise add a dichloromethane solution of boron tribromide (1 M) (0.03 mL, 0.03 mmol) at -78 °C. Stir for 2 h and then warm to room temperature. React overnight. Quench the reaction mixture by dropping it into saturated aqueous sodium bicarbonate solution (50 mL), and extract with EA (50 mL). Wash the EA phase with water (50 mL × 2), then wash with saturated aqueous sodium chloride solution (50 mL), and dry over anhydrous sodium sulfate to obtain the product IIc-1’ (3.6 mg, 0.01 mmol), an orange solid, with a yield of 90%. MS-ESI: m / z 362.1 [M+H] + 。

[0700] Step 4: Prepare the product II-25

[0701] Add IIc-1’ (3.6 mg, 0.01 mmol) and potassium carbonate (4 mg, 0.03 mmol) to N,N-dimethylformamide (DMF) (0.1 mL), dropwise add epifluorohydrin (1.5 μL, 0.02 mmol), and react at 80 °C for 4 h. After the reaction is complete, purify by high performance liquid chromatography (HPLC) (acetonitrile / water). The resulting product I-1 is a yellow solid with a yield of 7%. LC-MS (ESI) [M+H] + : 438.1。 1 H NMR (400 MHz, DMSO-d6) δ 8.98 (d, J = 2.4 Hz, 1H), 8.79 (s, 2H), 8.17 (dd, J = 8.2, 2.4 Hz, 1H), 7.94–7.86 (m, 2H), 7.84 (d, J = 8.3 Hz, 1H), 7.74 (d, J = 2.6 Hz, 1H), 7.63 (d, J = 15.9 Hz, 1H), 7.49 (s, 1H), 7.16 (dd, J = 8.9, 2.5 Hz, 1H), 4.64–4.53 (m, 1H), 4.51–4.41 (m, 1H), 4.15–4.04 (m, 3H), 2.87 (s, 3H).

[0702] Example II-26: Preparation of Compound II-26

[0703]

[0704] The synthesis method of Example II-26 is the same as that of Example II-25, except that 5-bromo-2-(methylamino)pyrimidine is replaced by 6-bromo-3-(methylamino)pyridazine. The resulting product II-26 is a yellow solid with a yield of 5%. LC-MS (ESI) [M+H] + : 438.1。 1HNMR(400MHz, DMSO-d6) δ 9.26 (d, J = 2.3 Hz, 1H), 8.42 (dd, J = 8.3, 2.4 Hz, 1H), 8.25 (d, J = 9.6 Hz, 1H), 8.00–7.87 (m, 3H), 7.75 (d, J = 2.5 Hz, 1H), 7.68 (d, J = 15.8 Hz, 1H), 7.36 (d, J = 9.5 Hz, 1H), 7.17 (dd, J = 8.9, 2.6 Hz, 1H), 4.63–4.53 (m, 1H), 4.51–4.42 (m, 1H), 4.18–4.02 (m, 3H), 3.05–2.98 (m, 3H).

[0705] Example II-27: Preparation of Compound II-27

[0706]

[0707] The synthesis method of Example II-27 is the same as that of Example II-25, except that 5-bromo-2-(methylamino)pyrimidine is replaced by 5-bromo-2-(methylamino)pyrazine. The obtained product II-27 is a yellow solid with a yield of 7%. LC-MS (ESI) [[M+H]] + : 438.1. 1 HNMR(400MHz, DMSO-d6) δ 9.22 (d, J = 2.3 Hz, 1H), 8.79–8.70 (m, 1H), 8.37 (dd, J = 8.3, 2.3 Hz, 1H), 8.07–8.04 (m, 1H), 7.93 - 7.79 (m, 3H), 7.74 (d, J = 2.6 Hz, 1H), 7.63 (d, J = 15.9 Hz, 1H), 7.43 (s, 1H), 7.16 (dd, J = 8.9, 2.6 Hz, 1H), 4.65–4.53 (m, 1H), 4.52 - 4.41 (m, 1H), 4.16 - 4.02 (m, 3H), 2.88 (s, 3H).

[0708] Example II-28: Preparation of Compound II-28

[0709]

[0710] Enrich 200 mCi 18 F on a QMA column and displace it with 0.7 mL of eluent (150 mg 222, 15 mg K2CO3, 9 mL acetonitrile, 1 mL water) 18F was rinsed into the reaction flask; high-purity nitrogen was introduced, and the solvent was evaporated to dryness by heating at 110 °C for 20 min; 0.2 mL of acetonitrile and 0.2 mg of 2-(oxiran-2-yl)methyl 4-methylbenzenesulfonate were added to the reaction flask, and the reaction was set at 90 °C for 10 min; the acetonitrile was evaporated to dryness by a rotary evaporator, 0.2 mL of DMF solvent and 0.5 mg of IIc-1' were added, and the reaction was carried out at 130 °C for 10 min; the mixture was separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 30:70, flow rate: 3 mL / min, elution time: 29 min), the product was collected and solid-phase extracted with a C18 column, and the final product II-28 was obtained by eluting with ethanol, and the labeling efficiency was 6%.

[0711] Example II-29: Preparation of Compound II-29

[0712]

[0713] The labeling method was as shown in Example II-28, except that the precursor IIc-1' was replaced with

[0714] Example III-1: Preparation of Compound III-1

[0715]

[0716] Synthetic route of Compound III-1:

[0717]

[0718] First step: Preparation of Intermediate 17

[0719] To a mixture of Compound 15 (1.280 g, 4.51 mmol), Compound 16 (1.5 g, 4.51 mmol), cesium carbonate (4.41 g, 13.53 mmol), palladium acetate (101 mg, 0.45 mmol) and 1,1'-bis(diphenylphosphino)ferrocene (DPPF, 249 mg, 0.45 mmol), 1,4-dioxane (15 mL), ethanol (5 mL) and water (5 mL) were added. The reaction mixture was reacted at 120 °C for 3 hours under a nitrogen atmosphere. Then, the reaction mixture was filtered and directly concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain white solid Compound 17 (1.2 g, yield 73.5%). LCMS: m / z (ESI) = 363.6 [M+H]+

[0720] Second step: Preparation of Intermediate 7

[0721] To a solution of compound 17 (1.2 g, 3.31 mmol) in dioxane (25 mL), bis(pinacolato)diboron (1.68 g, 6.62 mmol), Pd(dppf)Cl2 (242 mg, 0.33 mmol) and potassium acetate (973 mg, 9.93 mmol) were added. The reaction mixture was stirred at 80 °C for 15 h. After that, the reaction solution was filtered and concentrated directly, and then the residue was purified by column chromatography (petroleum ether: ethyl acetate = 3:1) to obtain gray solid compound 7 (400 mg, yield 29.4%).

[0722] Step 3: Preparation of intermediate 3

[0723] Compound 1 (5 g, 26.88 mmol) and compound 2 (4.8 g, 26.88) were dissolved in dimethyl sulfoxide (DMSO, 100 mL), and then 50% aqueous potassium hydroxide solution (100 mL) was added dropwise slowly, and the mixture was stirred overnight at room temperature. After the reaction was completed, a yellow precipitate was formed. The solid was collected by filtration and dried under reduced pressure to obtain white compound 3 (3.2 g, yield: 34% yield). LCMS: m / z (ESI) = 346.8 / 348.8 [M+H]+

[0724] Step 4: Preparation of intermediate 4

[0725] At -78 °C, 1 M boron tribromide (BBr3) solution in dichloromethane (1 M, 92.5 mL, 92.5 mmol) was added dropwise to a solution of compound 3 (3.2 g, 9.25 mmol) in dichloromethane (100 mL). After stirring at -78 °C for 2 h, the temperature was raised to room temperature and stirring was continued for 15 h. The reaction solution was poured into ice water (500 mL), and then saturated sodium bicarbonate solution was added dropwise slowly until the pH reached 8. The precipitate was collected by filtration to obtain yellow solid compound 4 (2.4 g, yield 78%). 1 HNMR (400 MHz, DMSO) δ 8.76 (d, J = 2.4 Hz, 1H), 8.12 (dd, J = 8.4, 2.4 Hz, 1H), 7.84 (d, J = 15.9 Hz, 1H), 7.80 (d, J = 8.8 Hz, 1H), 7.70 (t, J = 13.1 Hz, 1H), 7.51 (d, J = 15.9 Hz, 1H), 7.35 (d, J = 2.3 Hz, 1H), 6.97 (dd, J = 8.8, 2.4 Hz, 1H).

[0726] Step 5: Preparation of intermediate 6

[0727] To a solution of Compound 4 (2.4 g, 7.2 mmol) in N,N-dimethylformamide (DMF, 30 mL), cesium carbonate (Cs2CO3, 2.98 g, 21.6 mmol) and Compound 5 (2.74 g, 36 mmol) were added. The reaction mixture was heated to 80 °C and maintained for 4 hours. After cooling to room temperature, the reaction mixture was poured into ice water (100 mL), stirred for 10 min, and then filtered. The solid was collected and purified by flash column chromatography (dichloromethane:methanol = 10:1) to obtain yellow solid Compound 6 (1.5 g, yield 51%). LCMS: m / z (ESI) = 409.0 [M+H]+

[0728] Step 6: Preparation of Intermediate 8

[0729] To a mixture of Compound 6 (300 mg, 0.73 mmol), Compound 7 (300 mg, 0.73 mmol), Pd(dppf)Cl2 (51 mg, 0.07 mmol) and K2CO3 (294 mg, 2.16 mmol) were added 1,4-dioxane (10 mL) and water (2 mL). The reaction mixture was stirred at 80 °C for 15 hours under a nitrogen atmosphere. The reaction solution was filtered, and the filtrate was concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain yellow solid Compound 8 (360 mg, yield 81%). LCMS: m / z (ESI) = 613.2 [M+H]+

[0730] Step 7: Preparation of Compound III-1

[0731] To a solution of Compound 8 (360 mg, 0.59 mmol) in dichloromethane (10 mL), trifluoroacetic acid (5 mL) was added, and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated and purified by preparative high performance liquid chromatography (containing 0.05% trifluoroacetic acid) to obtain the trifluoroacetate salt of yellow solid Compound III-1 (60 mg, yield 16.7%). LCMS: m / z (ESI) = 513.2 [M+H]+. 11H NMR (400 MHz, DMSO) δ 9.12 (d, J = 2.0 Hz, 1H), 9.01 (d, J = 2.0 Hz, 1H), 8.37 - 8.3 (m, 2H), 8.04 (d, J = 8.8 Hz, 1H), 7.98 - 7.89 (m, 5H), 7.75 (d, J = 3.5 Hz, 1H), 7.68 (d, J = 16 Hz, 1H), 7.17 (dd, J = 8.8, 2.4 Hz, 1H), 6.69 (d, J = 7.2 Hz, 2H), 4.60 - 4.58 (m, 1H), 4.57 - 4.50 (m, 1H), 4.12 - 4.08 (m, 3H), 2.77 (s, 3H).

[0732] Example III - 2: Preparation of Compound III - 2

[0733]

[0734] Synthetic route of Compound III - 2:

[0735]

[0736] The first step: Preparation of Intermediate 3

[0737] Dissolve Compound 1 (10.00 g, 65.33 mmol) and Compound 2 (22.11 g, 78.40 mmol) in dioxane (80 mL), then add water (20 mL), Pd(dppf)Cl2 (1.00 g, 1.36 mmol) and K2CO3 (27.04 g, 196.00 mmol). Then stir the reaction mixture at 80 °C for 2 hours. Concentrate the reaction mixture and purify the residue by flash column chromatography on silica gel, using (28% EtOAc in PE) as the eluent to obtain yellow solid Compound 3 (5 g, yield 29.00%), LCMS: m / z (ESI) = 338.6 [M + H] +

[0738] The second step: Preparation of Intermediate 4

[0739] To a solution of compound 3 (5.00 g, 18.94 mmol) and compound 3A (14.58 g, 94.70 mmol) in toluene (50 mL) were added diisopropylethylamine (DIEA, 7.34 g, 56.82 mmol), tri-tert-butylphosphine tetrafluoroborate (1.09 g, 3.78 mmol), and Pd2(dba)3 (1.73 g, 1.89 mmol). The reaction mixture was then stirred at 100°C for 12 hours. The reaction mixture was then concentrated, and the residue was purified by flash column chromatography on silica gel using 30% EA in PE as the eluent to afford compound 4 (0.80 g, 12.46% yield) as a yellow solid. LCMS: m / z (ESI) = 338.6 [M+H] +

[0740] Step 3: Preparation of Compound III-2

[0741] To a solution of compound 4 (0.80 g, 2.36 mmol) and compound 5 (0.64 g, 2.83 mmol) in dioxane (8 mL) and water (2 mL) were added Pd(dppf)Cl2 (0.10 g, 0.14 mmol) and K2CO3 (0.97 g, 7.09 mmol). The reaction mixture was then stirred at 80°C for 2 hours. The reaction mixture was then concentrated to obtain a residue. The residue was purified by flash column chromatography on silica gel using 26% EtOAc in PE as the eluent to obtain the crude product. The crude product was then purified by preparative high-performance liquid chromatography (pre-HPLC) (instrument: Shimadzu LH-40 liquid handler, Shimadzu LC-20AP pump, Shimadzu SPD-20AP UV detector; column: Ultimate XB-C18, 50×250 mm×10 μm; mobile phase A: H 2 O with FA (10 mmol / L); mobile phase B: CH 3 CN; gradient: B increased from 12% to 42% in 30 minutes, then maintained at 100% B for 3 minutes; flow rate: 20 mL / min; retention time Rt = 15.0 minutes; column temperature: 30° C.; wavelength: 214 nM, 254 nM) to obtain yellow solid compound III-2 (58.1 mg, yield: 6.8%). LCMS: m / z (ESI) = 362.4 [M+H] +.1H NMR(400MHz,DMSO)δ9.96(s,1H),8.99(d,J=2.0Hz,1H),8.64(d,J=2.4Hz,1H),8.19-8.14(m,2H),7.87(d,J=16Hz,1H),7.82(d,J=8.8Hz,2H),7.59(d,J=16Hz,1H),7.40(d,J=2.4Hz,1H),7.00-6.96(m,2H),3.92(s,3H).

[0742] Example III-3: Preparation of Compound III-3

[0743]

[0744] Synthetic route of Compound III-3:

[0745]

[0746] To a mixture of Compound 8 (300 mg, 0.73 mmol), Compound 9 (300 mg, 140 mmol), Pd(dppf)Cl2 (51 mg, 0.07 mmol) and K2CO3 (294 mg, 2.16 mmol) was added 1,4-dioxane (10 mL) and water (2 mL). Under a nitrogen atmosphere, the reaction mixture was heated to 80 °C and maintained for 15 hours. The reaction solution was filtered and concentrated, and the residue obtained was purified by preparative high performance liquid chromatography (containing 0.05% formic acid) to give Compound III-3 as a gray solid (63.0 mg, 18.0%). LCMS: m / z (ESI) = 477.2 [M+H]+. 1H NMR (400 MHz, DMSO) δ 8.96 (d, J = 2.4 Hz, 1H), 8.58 (d, J = 6.8 Hz, 1H), 8.09 (dd, J = 8.2, 2.4 Hz, 1H), 7.96 (dd, J = 8.8, 2.4 Hz, 1H), 7.93-7.82 (m, 2H), 7.75 (dd, J = 14.4, 5.4 Hz, 2H), 7.62 (d, J = 15.8 Hz, 1H), 7.20-7.12 (m, 1H), 6.57 (d, J = 8.8 Hz, 1H), 5.51 (d, J = 9.2 Hz, 1H), 4.58 (d, J = 12.8 Hz, 1H), 4.47 (t, J = 8.2 Hz, 1H), 4.09 (d, J = 17.6 Hz, 3H), 3.46 (d, J = 6.6 Hz, 4H), 1.97 (t, J = 6.6 Hz, 4H).

[0747] Example III-4: Preparation of Compound III-4

[0748]

[0749] Synthetic route of Compound III-4:

[0750]

[0751] To a mixture of Compound 8 (300 mg, 0.73 mmol), Compound 10 (122 mg, 0.73 mmol), Pd(dppf)Cl2 (51 mg, 0.07 mmol) and K2CO3 (294 mg, 2.16 mmol) was added 1,4-dioxane (10 mL) and water (2 mL). The reaction mixture was heated to 80 °C under a nitrogen atmosphere and reacted for 15 hours. Then, the reaction mixture was filtered and concentrated, and the residue obtained was purified by preparative high performance liquid chromatography (containing 0.05% formic acid) to obtain yellow solid Compound III-4 (78.0 mg, yield 23.5%). LCMS: m / z (ESI) = 452.2 [M+H]+. 1H NMR (400 MHz, DMSO) δ 9.11 (d, J = 2.4 Hz, 1H), 8.38 - 8.31 (m, 3H), 8.12 (d, J = 8.8 Hz, 2H), 7.95 (t, J = 11.2 Hz, 3H), 7.72 (dd, J = 22.8, 9.2 Hz, 2H), 7.17 (dd, J = 8.8, 2.6 Hz, 1H), 4.61 - 4.43 (m, 3H), 4.13 - 4.05 (m, 3H).

[0752] Example III-5: Preparation of Compound III-5

[0753]

[0754] Synthetic route of Compound III-5:

[0755]

[0756] The first step: Preparation of Intermediate 2

[0757] At -78 °C, LDA (157 mmol, 157 mmol) was added to a solution of compound 1 (25.00 g, 142.90 mmol) in tetrahydrofuran (450 mL), and the reaction mixture was stirred at this temperature for 1 hour. Then, a solution of iodine (142.90 g, 142.90 mmol) in anhydrous tetrahydrofuran (50 mL) was added, and stirring was continued at -78 °C for 1.5 hours. At this temperature, the reaction mixture was quenched with saturated aqueous sodium thiosulfate solution and extracted with ethyl acetate (3 × 500 mL). The combined organic layers were washed with saturated brine (100 mL), dried over sodium sulfate, filtered, and concentrated to give a residue. The residue was purified by flash column chromatography on silica gel using (18% EtOAc in PE) as the eluent to give white solid compound 2 (16 g, yield 37.2%). 1H NMR (400 MHz, DMSO) δ 8.31 (t, J = 8.0 Hz, 1H), 7.43 - 7.41 (d, J = 8.0 Hz, 1H).

[0758] Step 2: Preparation of intermediate 4

[0759] At 25 °C, Pd(dppf)Cl2 (0.20 g, 0.273 mmol) and K2CO3 (2.75 g, 19.9 mmol) were added to a solution of compound 2 (2.00 g, 6.64 mmol) and compound 3 (2.66 g, 7.97 mmol) in dioxane (20 mL) and water (4 mL). Then, the reaction mixture was stirred at 80 °C for 2 hours. After concentration of the reaction mixture, a residue was obtained. The residue was purified by flash column chromatography on silica gel using (18% EtOAc in PE) as the eluent to give yellow solid compound 4 (1.1 g, yield 43.47%). LCMS: m / z (ESI) = 326.0 [M - 56 + H] +

[0760] Step 3: Preparation of intermediate 5

[0761] To a solution of compound 4 (1.10 g, 2.88 mmol) and compound 4A (2.22 g, 14.43 mmol) in toluene (20 mL) were added tri-tert-butylphosphine tetrafluoroborate (0.16 g, 0.57 mmol), Pd2(dba)3 (0.26 g, 0.28 mmol) and diisopropylethylamine (DIEA, 1.11 g, 8.64 mmol) at 25 ° C. The reaction mixture was then stirred at 100 ° C for 12 hours. The reaction mixture was concentrated to obtain a residue. The residue was purified by silica gel flash chromatography using (30% EtOAc in PE) as eluent to obtain yellow solid compound 5 (0.78 g, 59.54% yield). LCMS:, m / z (ESI) = 400.4 [M-56 + H] +

[0762] Step 4: Preparation of Intermediate 7

[0763] To a solution of compound 5 (0.78 g, 1.71 mmol) and compound 6 (0.47 g, 2.05 mmol) in dioxane (20 mL) and water (4 mL) was added Pd(dppf)Cl2 (0.20 g, 0.27 mmol) and K2CO3 (0.70 g, 5.13 mmol) at 25°C. The reaction mixture was then stirred at 80°C for 2 hours. The reaction mixture was concentrated to obtain a residue. The residue was purified by flash chromatography on silica gel using (22% EA in PE) as the eluent to obtain compound 7 (0.50 g, 61.72% yield) as a yellow solid. LCMS: m / z (ESI) = 478.6 [M+H]+.

[0764] Step 5: Preparation of compound III-5

[0765] Compound 7 (0.50 g, 1.04 mmol) was dissolved in HCl-dioxane solution (5 mL, 4 mmol / L) and stirred at 25 °C for 1 hour. The reaction mixture was concentrated to obtain the crude product. The crude product was purified by preparative high performance liquid chromatography (HPLC) (instrument: Shimadzu LH-40 liquid processor, Shimadzu LC-20AP pump, Shimadzu SPD-20AP UV detector; chromatographic column: Ultimate XB-C18, 50×250 mm×10 μm; mobile phase A: water containing formic acid (10 mmol / L); mobile phase B: acetonitrile; gradient: phase B was increased from 18% to 48% in 30 minutes, and then maintained at 100% phase B for 3 minutes; flow rate: 20 mL / min; retention time Rt = 12.5 min; column temperature: 30 °C; detection wavelength: 214 nM, 254 nM) to obtain the yellow solid compound III-5 (63.1 mg, yield: 16.1%). LCMS: m / z (ESI) = 379.4 [M+H] + 1H NMR (400 MHz, DMSO) δ 9.99 (s, 1H), 8.33 (s, 1H), 8.14–8.10 (m, 1H), 7.86–7.68 (m, 4H), 7.54 (d, J = 16 Hz, 1H), 7.40 (d, J = 2.4 Hz, 1H), 7.00–6.98 (m, 1H), 6.90–6.86 (m, 1H), 6.57 (d, J = 8.8 Hz, 1H), 2.83 (d, J = 4.8 Hz, 3H).

[0766] Example III-6: Preparation of Compound III-6

[0767]

[0768] Synthetic route of Compound III-6:

[0769]

[0770] Referring to the synthesis method of II-1 in Example II-1, 6-methoxy-2-methylbenzothiazole was replaced with II-1a to obtain the product III-6 (54 mg, yellow solid). LC-MS (ESI): [M+1]+ = 536.1.

[0771] Example III-7: Preparation of Compound III-7

[0772]

[0773] Synthetic route of Compound III-7:

[0774]

[0775] Step 1: Preparation of Intermediate II-2b

[0776] Dissolve 4-iodo-6-methoxypyridin-3-amine (5.10 g, 20.4 mmol) in dichloromethane (50 mL). Add triethylamine (4.27 mL, 30.6 mmol) and acetyl chloride (1.74 mL, 24.5 mmol) under ice-bath cooling, and then stir at room temperature for 1 hour. Wash the reaction mixture successively with water and saturated brine, and dry over anhydrous sodium sulfate. Evaporate the solvent under reduced pressure, wash the precipitated powder with isopropyl ether, and collect by filtration to obtain Intermediate II-2b as a white solid (4.42 g, yield 74%). LC-MS (ESI): [M+1]+ = 293.0.

[0777] Step 2: Preparation of Intermediate II-2c

[0778] At room temperature, dissolve II-2b (1.32 g, 4.51 mmol), potassium sulfide (1.49 g, 13.5 mmol), and copper(I) iodide (85.8 mg, 0.451 mmol) in DMF (25 mL). Heat the reaction solution to 90 °C and stir for 1 hour. Subsequently, quench the reaction solution with 1 N hydrochloric acid, extract with ethyl acetate, wash with saturated brine, dry over anhydrous sodium sulfate, and filter. Concentrate the filtrate under reduced pressure, and purify the resulting residue by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 2:1) to obtain II-2c (354 mg, yellow solid, yield 43.5%). LC-MS (ESI): [M+1]+ = 181.0.

[0779] For Steps 3, 4, and 5, refer to the synthesis method of II-1 in Example II-1, and replace 6-methoxy-2-methylbenzothiazole with II-2c to obtain Product III-7 (24 mg, yellow solid). LC-MS (ESI): [M+1]+ = 438.1.

[0780] Example III-8: Preparation of Compound III-8

[0781]

[0782] Synthetic route of Compound III-8:

[0783]

[0784] Referring to the synthesis method of II-1 in Example II-1, replace 6-methoxy-2-methylbenzothiazole with II-3a to obtain Product III-8 (28 mg, yellow solid). LC-MS (ESI): [M+1]+ = 439.1, and the synthesis of II-3a was carried out according to the method in Chem. Pharm. Bull. 1958, 6, 334-338.

[0785] Example III-9: Preparation of Compound III-9

[0786]

[0787] Synthetic route of Compound III-9:

[0788]

[0789] First step: Preparation of Intermediate II-4b

[0790] Under nitrogen protection, use a syringe to dissolve II-4a (2.49 g, 15 mmol) in dry tetrahydrofuran (70 mL), and inject it into a dry single-necked flask. Cool this solution to -78 °C, and then add 25 mL of tert-butyllithium (1.5 M) dropwise within 15 minutes. Then, let the temperature rise to 0 °C and stir for 2 hours. Cool the reaction mixture to -78 °C again, and then add iodine (9.52 g, 37.5 mmol, dissolved in 5 mL of dry tetrahydrofuran solution). Pour the reaction mixture into an aqueous ammonium chloride solution and extract with ethyl acetate. The organic phase is washed with a sodium thiosulfate solution, dried over magnesium sulfate, and concentrated under reduced pressure. The crude product is purified by silica gel chromatography (eluent: dichloromethane / methanol 100 / 0 - 95 / 5). Intermediate II-4b (1.5 g, yield 34%) is obtained. LC-MS (ESI): [M+1]+ = 292.9.

[0791] For the second, third, fourth, and fifth steps, referring to the synthesis method of III-7 in Example III-7, replace II-2b with II-4b to obtain Product III-9 (56 mg, yellow solid). LC-MS (ESI): [M+1]+ = 438.1.

[0792] Example III-10: Preparation of Compound III-10

[0793]

[0794] Synthetic route of Compound III-10:

[0795]

[0796] Referring to the synthesis method of III-7 in Example III-7, replace II-2a with II-5a to obtain the product III-10 (38 mg, yellow solid). LC-MS (ESI): [M+1]+ = 438.1.

[0797] Example III-11: Preparation of Compound III-11

[0798]

[0799] Synthetic route of Compound III-11:

[0800]

[0801] First step: Preparation of Intermediate II-6a

[0802] Dissolve 1a-1 (2.13 g, 10 mmol) in methanol (43 mL), then add potassium carbonate (2.76 g, 20 mmol) to the mixture, and then add a methanol (14 mL) solution of (1-diazo-2-oxo-propyl)-phosphonic acid dimethyl ester (2.14 g, 11 mmol) at room temperature, and stir the resulting mixture for 2 h. Subsequently, pour the mixture into 1 M sodium carbonate solution, extract with ethyl acetate, combine the organic matters, wash with saturated brine, dry with sodium sulfate, filter and concentrate. After separation and purification by silica gel column chromatography, Intermediate II-6a (1.29 g, 62%) is obtained as a yellow liquid. LC-MS (ESI): [M+1]+ = 210.1.

[0803] Second step: Preparation of Intermediate II-6c

[0804] In a 5 mL microwave tube, add II-6a (142.3 mg, 0.68 mmol), II-6b (197.9 mg, 0.68 mmol), tetrakis(triphenylphosphine)palladium (40 mg, 0.034 mmol), copper(I) iodide (0.02 g, 0.05 mmol) and triethylamine (0.28 mL, 2.04 mmol), and the solvent is acetonitrile (2.0 mL). Irradiate the suspension in a microwave reactor at 100 °C for 5 minutes. After cooling to room temperature, evaporate the solvent under vacuum. The residue is purified by silica gel column chromatography using n-hexane:dichloromethane (1:0 - 0:1) as the eluent to obtain II-6c (53 mg, 21%) as a yellow solid. LC-MS (ESI): [M+1]+ = 373.1.

[0805] The third and fourth steps refer to the synthesis method in Example III-6 to obtain the product III-11 (15 mg, yellow solid). LC-MS (ESI): [M+1]+ = 435.1.

[0806] Example III-12: Preparation of Compound III-12

[0807]

[0808] Synthetic route of Compound III-12:

[0809]

[0810] Referring to the synthesis method of II-1 in Example II-1, replace 5-bromo-2-(methylamino)pyridine with II-7b to obtain the product III-12 (63 mg, yellow solid). LC-MS (ESI): [M+1]+ = 454.2.

[0811] Example III-13: Preparation of Compound III-13

[0812]

[0813] Synthetic route of Compound III-13:

[0814]

[0815] First step: Preparation of Intermediate II-8b

[0816] Mix II-8a (4.68 g, 20 mmol), piperazine (7.83 mL, 100 mmol), copper(I) iodide (76.18 mg, 0.4 mmol), 2-[(2,6-dimethylphenyl)amino]-2-oxoacetic acid (386 mg, 2 mmol), and sodium carbonate (5.3 g, 50 mmol). Add N,N-dimethylformamide (35 mL) to a three-necked flask and displace the air with nitrogen. Heat the temperature to 100 °C for reaction. After the reaction is completed, concentrate the reaction solution and then separate it by column chromatography. Obtain Intermediate II-8b (1.5 g, 39%). LC-MS (ESI): [M+1]+ = 193.1.

[0817] Second step: Preparation of Intermediate II-8c

[0818] Under a nitrogen atmosphere, dissolve II-7d (358 mg, 2 mmol), II-8b (375 mg, 1.95 mmol), and triethyl orthoformate (2.49 mL, 15 mmol) in methanol (12 mL). Heat the reaction mixture to 70 °C for 18 hours. After cooling to room temperature, filter out the precipitate and wash it with ether. The obtained solid is dried under high vacuum at room temperature. Obtain Intermediate II-8c (119 mg, yield 16%). LC-MS (ESI): [M+1]+ = 382.2.

[0819] For the third and fourth steps, refer to the synthesis method in Example III-6 to obtain Product III-13 (23 mg, gray solid). LC-MS (ESI): [M+1]+ = 444.2.

[0820] Example III-14: Preparation of Compound III-14

[0821]

[0822] The synthesis method of Compound III-14 refers to the synthesis method in Example II-1. Replace 5-bromo-2-(methylamino)pyridine with 2-hydroxy-5-bromopyridine to obtain Product III-14 (56 mg, brownish-gray solid). LC-MS (ESI): [M+1]+ = 424.1.

[0823] Example III-15: Preparation of Compound III-15

[0824]

[0825] The synthesis method of Compound III-15 refers to the synthesis method in Example II-1. Replace 5-bromo-2-(methylamino)pyridine with 5-bromo-2-cyanopyridine to obtain Product III-15 (35 mg, yellow solid). LC-MS (ESI): [M+1]+ = 433.1.

[0826] Example III-16: Preparation of Compound III-16

[0827]

[0828] The synthesis method of Compound III-16 refers to the synthesis method in Example II-1. Replace 6-methoxy-2-methylbenzothiazole with 2-methyl-benzothiazole-6-carboxylic acid to obtain Product III-16 (78 mg, yellow solid). LC-MS (ESI): [M+1]+ = 389.1.

[0829] Example III-17: Preparation of Compound III-17

[0830]

[0831] Synthesis route of Compound III-17:

[0832]

[0833] First step: Prepare Intermediate II-12c

[0834] At room temperature, II-12a (114.1 mg, 0.5 mmol), N-methylpiperazine (75 mg, 0.75 mmol) and K3PO4 (425 mg, 2.0 mmol) were added to a dry tetrahydrofuran (5 mL) solution, and then the reaction mixture was purged with nitrogen for 15 minutes. Then Ruphos Pd G1 (81.7 mg, 0.1 mmol) was added. The reaction mixture was continuously purged with argon for 5 minutes and then stirred at 80 °C in a sealed tube for 24 hours. After the reaction was completed, it was filtered through a celite pad, and the celite pad was washed with ethyl acetate (50 mL). The filtrate was concentrated under reduced pressure. It was further separated by column chromatography. Intermediate II-12c (28.5 mg, yield 23%) was obtained. LC-MS (ESI): [M+1]+ = 248.1.

[0835] Step 2: Preparation of compound III-17

[0836] Intermediate 1a-1 (24.6 mg, 0.115 mmol) and 11-12c (28.5 mg, 0.115 mmol) were dissolved in dimethyl sulfoxide (2 mL), and an aqueous solution of 50% potassium hydroxide (1 mL) was slowly added dropwise, and the mixture was stirred at room temperature overnight. After the reaction was complete, a yellow solid precipitated. It was filtered, and the filter cake was washed with water. The solid was collected and dried to obtain product III-17 (35 mg, yellow solid, yield 68%). LC-MS (ESI): [M+1]+ = 443.2.

[0837] Example III-18: Preparation of compound III-18

[0838]

[0839] The synthesis method of compound III-18 was referred to the synthesis method in Example III-17, and N-methylpiperazine was replaced with 4-methylpiperazin-2-one to obtain product III-18 (12 mg, yellow solid). LC-MS (ESI): [M+1]+ = 457.2.

[0840] Example III-19: Preparation of compound III-19

[0841]

[0842] Synthetic route of compound III-19:

[0843]

[0844] Step 1: Preparation of intermediate II-14c

[0845] At room temperature, 3-fluoropropanoic acid II-14b (35.5 mg, 0.386 mmol) was dissolved in dichloromethane (4.0 mL), and N,N,N’,N’-tetramethyl-O-(7-azabenzotriazol-1-yl) hexafluorophosphate (293 mg, 0.77 mmol), N,N-diisopropylethylamine (127 μL, 0.77 mmol) and II-14a (63.4 mg, 0.386 mmol) were added. The reaction was carried out at room temperature for 40 minutes. After completion of the reaction, it was diluted with dichloromethane (100 mL), and the organic phase was washed successively with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 5 / 95). A pale yellow oil II-4c (74 mg, yield: 80%) was obtained. LC-MS (ESI): [M+1]+ = 239.1

[0846] For the second step, referring to the synthesis method in Example III-17, II-12c was replaced with II-14c to obtain the product III-19 (24 mg, yellow solid). LC-MS (ESI): [M+1]+ = 434.2.

[0847] Example III-20: Preparation of Compound III-20

[0848]

[0849] The synthesis of Compound III-20 was carried out referring to the synthesis method in Example II-1, replacing 5-bromo-2-(methylamino)pyridine with 7-bromo-N,N-dimethylnaphthalen-2-amine to obtain the product III-20 (5.6 mg, gray solid). LC-MS (ESI): [M+1]+ = 500.2.

[0850] Example III-21: Preparation of Compound III-21

[0851]

[0852] The synthesis of Compound III-21 was carried out referring to the synthesis method in Example III-17, replacing N-methylpiperazine with 2-fluoroethylamine hydrochloride to obtain the product III-21 (34 mg, yellow solid). LC-MS (ESI): [M+1]+ = 406.2.

[0853] Example III-22: Preparation of Compound III-22

[0854]

[0855] Synthetic route of Compound III-22:

[0856]

[0857] Step 1: Preparation of Intermediate II-17b

[0858] N-Bromosuccinimide (NBS, 585 mg, 3.29 mmol) was added to a solution of II-17a (323 mg, 3.29 mmol) in dimethylformamide (DMF, 17 mL). The reaction mixture was stirred at 30 °C for 1 hour. Then the reaction solution was concentrated and purified by flash silica gel chromatography (the proportion of methanol in ethyl acetate was increased from 0% to 20%) to obtain II-17b as a yellow solid (286 mg, yield 49%). LC-MS (ESI): [M+1]+ = 176.9.

[0859] Step 2: Preparation of Intermediate II-17d

[0860] II-17b (286 mg, 1.62 mmol), II-17c (244.5 mg, 1.62 mmol), 1,1-bis(diphenylphosphino)ferrocene dichloropalladium (118.5 mg, 0.162 mmol) and potassium carbonate (670 mg, 4.85 mmol) were dissolved in 1,4-dioxane (10 mL) and water (3 mL). After protecting with nitrogen, the reaction was carried out at 80 °C overnight. After the reaction was complete, it was purified by flash silica gel column chromatography, petroleum ether (PE) / ethyl acetate (EA), to obtain the product II-17d (178 mg, yield 54%). LC-MS (ESI): [M+1]+ = 204.1.

[0861] The third, fourth, and fifth steps were carried out according to the synthesis method in Example II-1, replacing 1a-1 with II-17d and replacing epifluorohydrin with fluoroethyl p-toluenesulfonate (II-17f) to obtain the product III-22 (59 mg, yellow solid). LC-MS (ESI): [M+1]+ = 397.1.

[0862] Example III-23: Preparation of Compound III-23

[0863]

[0864] Synthetic route of Compound III-23:

[0865]

[0866] Step 1: Synthesis of Precursor III-X

[0867] Id-38 (0.06 mmol), If-1 (0.29 mmol) and potassium carbonate (16 mg, 0.12 mmol) were added to DMF (0.5 mL), and the reaction was carried out at 80 °C for 2 h. After the reaction was complete, it was purified by flash silica column chromatography (PE / EA) to obtain product III-X (3.8 mg, 0.006 mmol), a yellow solid, with a yield of 57%. MS-ESI: m / z 703.2 [M+H] + 。

[0868] Step 2: Synthesis of precursor III-23

[0869] 150 mCi 18 of 18 F was enriched on a QMA column, and F was eluted with 0.6 mL of eluent (150 mg of Kryptofix 222, 15 mg of K2CO3 dissolved in 9 mL of acetonitrile and 1 mL of water) into the reaction flask; under high-purity nitrogen, the solvent was dried by heating at 110 °C for 20 min; 0.5 mg of I-X was added to the reaction flask and reacted at 140 °C for 10 min; then 0.2 mL of 2N hydrochloric acid was added and reacted at 105 °C for 8 min to remove the THP protecting group; the reaction solution was separated and purified by HPLC (HPLC conditions: CAPCELL PAK C18 UG80 10 mm×250 mm, acetonitrile: water = 25:75, flow rate: 3 mL / min), the product was collected and solid-phase extracted with a C18 column, and the final product III-23 was obtained by eluting with ethanol, and the labeling efficiency was 14%.

[0870] Reference Example 1: Preparation of reference compound 1

[0871]

[0872] Synthetic route of reference compound 1:

[0873]

[0874] Reference compound 1

[0875] Step 1: Preparation of intermediate 13

[0876] To a solution of Compound 11 (4 g, 25.64 mmol) and Compound 12 (3.6 g, 25.64 mmol) in 1,4-dioxane (100 mL) and water (20 mL) were added Pd(dppf)Cl2 (220 mg, 0.3 mmol) and K2CO3 (10.46 g, 76.92 mmol). The reaction mixture was heated to 80 °C under nitrogen protection and reacted for 15 hours. After the reaction was completed, the reaction mixture was filtered and concentrated. The residue was purified by column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain yellow solid Compound 13 (800 mg, 14.3% yield). 1H NMR (400 MHz, CDCl3) δ 9.90 (s, 1H), 7.73 (d, J = 4.0 Hz, 1H), 7.69 - 7.59 (m, 1H), 7.49 (t, J = 8.0 Hz, 1H), 7.04 (d, J = 6.8 Hz, 1H), 6.40 (d, J = 8.4 Hz, 1H), 2.99 (d, J = 4.0 Hz, 3H).

[0877] Step 2: Preparation of Reference Compound 1

[0878] Compound 13 (600 mg, 2.75 mmol) and Compound 14 (836 mg, 2.75 mmol) were dissolved in methanol (50 mL) and heated to 80 °C for reaction for 15 hours. After the reaction was completed, it was cooled to room temperature and filtered. The solid was recrystallized with methanol and washed, and after drying, yellow solid Reference Compound 1 (60 mg, 4.3%) was obtained. LCMS: m / z (ESI) = 378.2 [M+H]+. 1H NMR (400 MHz, DMSO) δ 8.45 - 8.41 (m, 2H), 8.28 (d, J = 8.6 Hz, 1H), 7.99 (d, J = 3.8 Hz, 1H), 7.87 (d, J = 15.2 Hz, 2H), 7.82 - 7.70 (m, 2H), 7.49 (t, J = 7.8 Hz, 1H), 7.20 (d, J = 7.4 Hz, 1H), 6.74 (d, J = 4.6 Hz, 1H), 6.49 (d, J = 8.4 Hz, 1H), 4.95 (d, J = 7.2 Hz, 2H), 2.88 (d, J = 4.8 Hz, 3H), 1.48 (t, J = 7.2 Hz, 3H).

[0879] Reference Example 2: Preparation of Reference Compound 2

[0880]

[0881] The synthesis of Reference Compound 2 was carried out with reference to the published patent KR20190090448. Reference Compound 2 was a yellow solid (61.7 mg), LCMS: m / z (ESI) = 435.6 [M+H]+ ..

[0882] Biological Test Example 1: Protein Level Activity Test

[0883] Fluorescence method and surface plasmon resonance (SPR) are commonly used experiments to detect the interaction between small molecules and proteins. The applicant used these two methods to test the binding affinity of the example compounds to α-synuclein.

[0884] 1.1 Experimental Method:

[0885] (1) Preparation of α-syn:

[0886] Co-transform α-syn plasmid and yeast N-acetyltransferase complex B plasmid into BL21(DE3) E. coli competent cells. Spread the transformation solution on a solid culture plate containing ampicillin and chloramphenicol (Gibco, catalog number 15140148) and incubate overnight at 37°C for plasmid transformation. Pick a single colony and place it in 20 ml of 2×YT medium (Sigma-Alcrich, 2× yeast extract tryptone medium, containing the corresponding penicillin and chloramphenicol), incubate overnight at 37°C and 220 rpm. Then inoculate the bacterial solution into 1 L of 2×YT medium at a volume ratio of 1:100. After 1 hour (when the OD600 of the cultured bacterial solution is 0.8 - 1), add 1 mM IPTG (isopropyl β-D-1-thiogalactopyranoside) and induce at 37°C for 4 h. Finally, collect Escherichia coli by centrifugation (4°C, 4500 rpm, 20 min). Resuspend the collected Escherichia coli with 60 - 80 ml of bacterial lysate (100 mM Tris-HCl, pH 8.0, 1 mM EDTA, 1 mM PMSF). Then use high-pressure crushing for bacterial lysis (4°C, 10 min); centrifuge the cell lysate (4°C, 15000 rpm, 25 min) and take the supernatant to boil in a water bath for 15 min; centrifuge again (4°C, 15000 rpm, 25 min) and add streptomycin sulfate (20 mg / ml, wt / vol) to the supernatant. Place the solution in a 4°C environment and stir continuously for 30 min; further centrifuge the lysate (4°C, 15000 rpm, 25 min) and adjust the pH of the supernatant to 3.5; finally, centrifuge (4°C, 15000 rpm, 25 min) and dialyze the supernatant overnight (4°C) (dialysis solution: 25 mM Tris-HCl, pH 8.0).

[0887] Purification was first carried out using an anion exchange column (Q column): First, wash the Q column with Buffer B (25 mM Tris-HCl, pH 8.0, 1 M NaCl), and equilibrate the Q column with Buffer A (25 mM Tris-HCl, pH 8.0) (5 - 10 column volumes). After filtering the protein dialysis solution through a 0.22 μm filter membrane, use a protein purification system to load the sample onto the Q column, and then wash away the unbound protein with Buffer A; set a gradient elution of 0 - 60% Buffer B for 40 min; use an automatic sample collection system to collect the eluate. Then, purification was carried out using a gel size exclusion column (Superdex 75): Collect the target protein solution eluted from the Q column, concentrate and filter it, and then load the sample using a sampling loop; elute the molecular sieve with DPBS buffer (Thermo Fisher, catalog number 14040117), and use an automatic sample collection system to collect the eluate. A pure α-synuclein monomer was prepared.

[0888] (2) Preparation method of α-synuclein aggregates: Place the α-syn monomer (200 μM, in 50 mM Tris, pH 7.5, 150 mM KCl buffer) on a ThermoMixer shaker and incubate at 37 °C and 900 rpm for 5 - 7 days to form amyloid fiber structures. Finally, use transmission electron microscopy (TEM) to detect the morphology of mature α-syn fibers.

[0889] (3) SPR test method: Wash the CMD 500 chip (GE) with double-distilled water, air-dry it, and then run the "d℃k" program to enter the Biacore T200 instrument. Remove impurities through the "desorb" program and run the "Prime" program to remove air bubbles. Dilute the incubated α-syn aggregates (5 mg / mL) with NaAc buffer at pH 4.0, 4.5, 5.0, and 5.5 to 20 μg / mL samples, inject them sequentially for 60 s, and elute with NaOH after injection to select the optimal coupling buffer according to the coupling amount. Inject 200 μL of the chip activation solution for 900 s to activate the carboxyl groups on the chip. Dilute the α-syn aggregates with NaAc buffer (pH = 4.0) to a 50 μg / mL protein sample, and then inject the protein sample for 110 s, repeating 10 times to allow the protein to fully couple with the chip. Inject the blocking solution (GE original protein blocking solution) for 900 s to block the unreacted carboxyl groups in the chip. Dilute the small molecule to be tested to different concentrations, run the "Kinetic" program in Biacore to inject samples sequentially, and use the system software to export the binding force and kinetic spectra of the small molecule.

[0890] (4) Fluorescence-based protein level binding assay: The small molecule was prepared into a 10 mM stock solution with DMSO and then diluted to 20 μM with PBS, followed by 7 serial dilutions (triple dilution each time); 30 μL of the test compound was added to a 384-well plate. In the experimental group, 30 μL of α-synuclein aggregates (Series of Example I and Example III:
μM; Series of Example II: 3 μM) was added, and in the control group, an equal volume of PBS was added. The 384-well plate was incubated at room temperature with shaking (50 rpm) for 1 hour; the maximum absorption and emission wavelengths of the small molecule were detected with a microplate reader, and the fluorescence value was detected at this wavelength. The fluorescence change values at different concentrations of the molecule were calculated by subtracting the control group from the experimental group, and the binding affinity of the small molecule to the protein was calculated using the Saturation binding module of GraphPad Prism.

[0891] 1.2 Experimental results:

[0892] Table 1 Test results of the α-synuclein aggregate binding ability of the test compounds

[0893]

[0894]

[0895] Note: A represents a K d value less than 0.01 - 0.5 μM, B represents a K d value between 0.5 and 2 μM, C represents a K d value between 2 and 10 μM, D represents a K d value greater than 10 μM, ND represents not tested.

[0896] Biological test example 2: Immunofluorescence staining of α-synuclein aggregated primary neurons

[0897] 2.1 Experimental method:

[0898] (1) Primary neuron culture: The glass slides were placed in a 24-well plate, and polylysine solution was added. SD rats at 15 - 18 days of pregnancy were dissected, and the left and right cerebral cortices of the fetal rats were taken and placed in HBSS buffer containing HEPES. After the fetal rat cortical tissue was rinsed with HBSS buffer, the brain tissue was digested with activated papain and DNase and placed in a 37 °C cell culture incubator, gently flipped several times every 5 minutes. After digestion, it was filtered through a 40 μm cell filter, and the filter mesh was rinsed with 15 ml of plating medium (DMEM (without pyruvate) with 10% FBS and 1% PS). The cell solution was centrifuged at 900 rpm for 5 minutes, resuspended with plating medium, and then cell counting was performed. At 10 - 15 × 10 4Plate with cells at a density of 1 cell / well and 0.5 ml of plating medium per well; after culturing in a cell incubator for 1 - 2 h, replace the medium with Neurobasal medium (Neurobasal medium containing 1% PS, B27 supplement, and 0.5 mM GlutaMAX), 1 ml per well. Finally, place it in a cell incubator to culture neurons until they mature (7 - 10 days).

[0899] (2) Cell treatment: After culturing primary neurons in vitro for 7 - 10 days, add α-syn PFF with a final concentration of 100 nM to the medium. The treated neurons are then cultured for another 30 d, and the neurons are collected for sample preparation.

[0900] (3) Immunofluorescence staining: Aspirate the cell culture medium, wash three times with PBS, then add 0.3% Triton X-100 and incubate for 10 min; after washing with PBS, add 10% goat serum to block for 1 h; after washing with PBS, add p-129-α-syn antibody (1:600, ab51253, Abcam) and incubate overnight at 4 °C; after washing with PBS, add secondary antibodies (1:1000, goat-anti-rabbit Alex Fluor 594 and goat-anti-mouse Alex Fluor 488, Invitrogen) and incubate for 2 h at room temperature; finally, add the test compound and incubate for 1 h at room temperature, wash with PBS, mount the slides, and take pictures using a fluorescence microscope or a laser confocal microscope (Olympus laser scanning confocal microscope FV3000).

[0901] 2.2 Experimental results:

[0902] The results of immunofluorescence staining are as Figure 1 shown. The compounds in this application series have green autofluorescence themselves. The experimental results show that the fluorescence of the test compound can co-localize with the fluorescence signal of the pS129 antibody (pathological α-syn), indicating that the compounds in this application can bind to α-syn aggregates in primary neurons, that is, the compounds in this application can detect α-syn aggregates in primary neurons.

[0903] Biological Test Example 3: Immunofluorescence Staining of PFF Mouse Brain Slices

[0904] 3.1 Experimental method:

[0905] (1) Construction of the fibril (PFF) mouse model (i.e., the PD mouse model): Weigh the mice and calculate the injection dose of α-syn PFF (0.2 μg / g). Anesthetize the 6-week-old mice and fix them on the stereotaxic apparatus. Adjust the position of the mouse brain to make it symmetric left and right. Cut the hair on the mouse's head and cut open the scalp to expose the skull. Under the microscope, combine with the micro-reader on the stereotaxic apparatus to adjust the mouse brain to make it flat and symmetric in the front, back, left, and right. Use a cranial drill to make holes in the dorsal striatum (dSTR), and then use a micro-injection pump to inject α-syn PFF and PBS into the bilateral brain regions of the mice respectively. The PD mice are cultured for 6-9 months after the operation, and then the mice are sacrificed and the brain tissues are obtained by dissection.

[0906] (2) Brain slice preparation: Fix the brain tissue with 4% paraformaldehyde solution overnight, dehydrate it with sucrose solution, embed it with a tissue freezing embedding machine, and use a slicer (Leica CM1860) to cut the brain tissue into 30-μm-thick slices. Collect the slices in a cryoprotectant solution (30% sucrose (w / v), 30% ethylene glycol (v / v) dissolved in PBS) and store them at -20 °C for subsequent fluorescence staining and imaging.

[0907] (3) Immunofluorescence staining: Wash the prepared brain slices three times with PBS, add 0.3% Triton X-100 and incubate for 10 min; wash three times with PBS, add 10% goat serum and block for 1 h; wash three times with PBS, add the p-129-α-syn antibody (1:600, ab51253, Abcam) and incubate overnight at 4 °C; wash three times with PBS, add the secondary antibody (1:1000, goat-anti-rabbit AlexFluor 594 and goat-anti-mouse Alex Fluor 488, Invitrogen) and incubate for 2 h at room temperature; wash three times with PBS, add the test compound and incubate for 1 h at room temperature; wash three times with PBS, mount the slices and observe with a fluorescence microscope or a laser confocal microscope.

[0908] 3.2 Experimental results:

[0909] The experimental results are as Figure 2 shown. The autofluorescence signal of the preferred compound (also known as the tracer) of this application co-localizes with the signal of the pS129 antibody (pathological α-syn), and there is no other non-specific binding signal, indicating that the compound of this application can specifically recognize the pathological α-syn aggregates in the brain tissue, verifying the ability of the compound of this application to bind to α-syn aggregates in the brain tissue.

[0910] Biological test example 4: Immunofluorescence staining of brain slices from PD patients

[0911] 4.1 Experimental method:

[0912] An Asian female PD patient, 86 years old, with a disease duration of 14 years, had brain tissue obtained by autopsy within 12 hours after her death. The immunofluorescence staining method was the same as that applied to the mouse brain slices in Biological Test Example 3.

[0913] 4.2 Experimental results:

[0914] The experimental results ( Figure 3 ) showed that the autofluorescence signal of the compound (also known as the tracer) of this application co-localized with the α-syn antibody signal, indicating that the compound of this application could label the pathological α-syn in the patient's brain slices. Combining the above immunofluorescence staining results of cells and mouse brain slices, it shows that the compound of this application has good target binding ability in vitro.

[0915] Biological Test Example 5: Ex-vivo staining experiment of PFF mouse brain slices

[0916] 5.1 Experimental method:

[0917] Using the method described in Biological Test Example 3, a PFF mouse model was constructed. 1 mg / kg of the test compound was injected into the mice via the tail vein. After 2 hours, the mice were sacrificed and the brain tissues were taken out to prepare brain slices and perform immunofluorescence staining. 5.2 Experimental results:

[0918] The experimental results are as Figure 4 shown. The compound (also known as the tracer) of this application could cross the blood-brain barrier of the mice and specifically recognize the pathological α-syn in the mouse brain, indicating its good drug-likeness in vivo.

[0919] Biological Test Example 6: Comparison of the binding activities of the compound to α-Syn, Aβ and Tau

[0920] 6.1 Test compounds

[0921] Compound of Example II-1, Reference Compound 1, Reference Compound 2, and Prior Art Compound C0505: (Prepared according to WO2014 / 097474)

[0922] 6.2 Experimental method

[0923] 1) The preparation method of α-synuclein aggregates refers to 1(1) of Activity Test Example 1.

[0924] 2) Expression and purification of human Tau protein

[0925] The genes encoding human Tau protein (containing amino acid sequences 266–391 (3R) and 297–391) were inserted into the pRK172 vector respectively. The plasmids were transformed into competent Escherichia coli BL21(DE3) cells. After culturing at 37 °C until the OD600 reached 0.8 - 1.2, 1 mM IPTG (isopropyl β-D-1-thiogalactopyranoside) was added, and the expression was induced at 24 °C for 16 hours. The cells were lysed in washing buffer (50 mM MES, pH 6.0, 10 mM EDTA, 10 mM DTT, 0.1 mM PMSF), followed by centrifugation and filtration. After purification of the protein through an SP column, it was precipitated with ammonium sulfate, and the precipitate was resuspended in phosphate buffer (pH 7.2 - 7.4) containing DTT and further purified through a Superdex 75 gel filtration column. Each fraction was analyzed by SDS-PAGE, and the protein concentration was measured using a nanospectrometer (Winner801 type). The purified protein was stored in phosphate buffer (pH 7.4) containing DTT, quickly frozen, and stored at -80 °C.

[0926] 3) Preparation of Tau and Aβ fibrils

[0927] Preparation of Tau fibrils: 190 μM Tau monomers (commercially available, abcam, catalog number ab246003) were dissolved in a buffer containing 10 mM phosphate buffer (PB, pH 6.0), 10 mM DTT, 200 mM MgCl2, and 0.02% NaN3. Incubation was carried out in a Corning 96-well black polystyrene microplate (Thermo) at 37 °C with orbital shaking at 200 rpm for 48 hours using a FLUOstar Omega microplate reader (BMG LABTECH).

[0928] Preparation of Aβ fibrils: 200 μM Aβ(1 - 40, E22Δ) peptide was dissolved in a buffer containing 10 mM phosphate buffer (PB, pH 7.4), 100 mM NaCl, and 0.05% NaN3. It was shaken at 700 rpm in a ThermoMixer and incubated at 37 °C for 7 days.

[0929] 4) Fluorescence binding experiment

[0930] 1 μM of the test compound was mixed with 1 μM of the fibrils, added to a buffer containing 50 mM Tris (pH 7.5) and 150 mM KCl, and incubated at 25 °C for 1 hour. Subsequently, the fluorescence signal of the mixture was measured using a microplate reader. Meanwhile, a control reaction without fibrils was set up to evaluate non-specific fluorescence. The experimental results are shown in the following table.

[0931] 6.3. Experimental Results

[0932] Kd (μM) of the binding activities of representative compounds to α-Syn, Aβ, and Tau

[0933]

[0934]

[0935] Compared with other reference compounds, the binding activity of the compound of Example II-1 of this application to α-syn is significantly better than that of Aβ and Tau, indicating that the compound of the present invention has a highly selective binding activity to α-syn.

[0936] Activity Test Example 7: Autoradiography (ARG) of mouse and PD patient brain slices

[0937] 7.1 Preparation of mouse model

[0938] Male C57BL / 6J mice (2 months old, 18 - 20 g, purchased from Shanghai Lingchang Biotechnology Co., Ltd.), 5 mice per cage, 12-hour light-dark cycle, free access to food and water. The mice were anesthetized with a mixture of 0.5% isoflurane and 1% oxygen, and the body temperature was maintained with a heating pad. α-Syn PFF (5 μg) was stereotaxically injected into the dorsal striatum (dSTR) of both hemispheres, with the coordinates: AP ± 2.0 mm, ML + 0.2 mm, DV - 2.60 mm. The first 0.5 μL was infused at a rate of 0.5 μL / min using a 10 μL microsyringe, and then the remaining 2 μL was infused at a rate of 0.2 μL / min. Control animals received sterile PBS in the same manner. The mice were monitored regularly after postoperative recovery.

[0939] 7.2 Autoradiography (ARG)

[0940] Prepare brain slices of the mouse model in 7.1 according to the method of Biological Test Example 3, and prepare brain slices of PD patients according to the method of Biological Test Example 4. Pre-incubate the brain slices in PBS buffer for 30 minutes at 25 °C, and then incubate them with 10 nM 18F-FD4 (Compound of Example II-8) in PBS buffer containing 20% ethanol at 25 °C for 1 hour. After rinsing with pre-cooled PBS buffer containing 20% ethanol, dry the brain slices and place them on an imaging plate, cover with a storage phosphor screen overnight. Then scan the plate using a high-sensitivity imaging plate scanner (CR-35Bio plus), and analyze the images using AutoRAD morphological quantitative analysis software. Exclude the non-specific and off-target binding of 18F-FD4 by adding excessive unlabeled FD4 (Compound of Example II-1) (10 μM for mouse brain, 2 μM for human brain) and MAO-A / B inhibitors (Clorgyline / Selegiline, 10 μM) respectively.

[0941] 7.3 Experimental Results

[0942] This experiment further evaluated the sensitivity of 18F-FD4 to α-syn aggregates in mouse and human brain slices. The results are as Figure 5A and 5B shown. At a concentration of 10 nM, 18F-FD4 produced significant radio signals in brain slices of α-syn PFF-injected mice and Parkinson's disease (PD) patients; while no signals were detected in brain slices of healthy individuals. Referring to the experimental method of Biological Test Example 3.1, it was confirmed by antibody immunostaining (β-amyloid antibody: NAB228, Cell Signaling Technology; Tau antibody: AT8, BioLegend; α-syn antibody: ab138501, ab51253, Abcam) that the positive radio signals co-localized with α-syn aggregates, but not with Aβ and Tau aggregates. After adding excessive unlabeled FD4, the radio signals completely disappeared, excluding the non-specific binding of 18F-FD4. In addition, after adding MAO-A and MAO-B inhibitors, the radio signals remained unchanged, excluding the possible non-specific targeting of monoamine oxidase (MAO) binding. These results support the potential of FD4 as a PET imaging tracer with high sensitivity and high selectivity for α-syn pathological aggregates.

[0943] Activity Test Example 8: PET Imaging of the Compounds of the Present Application in Rodents, Marmosets and Human Subjects

[0944] 8.1 Preparation of Rat Model

[0945] Sprague Dawley female rats (10 - 12 weeks old, 250 - 350 g, purchased from Shanghai Lingchang Biotechnology Co., Ltd.) were housed three per cage under a 12-hour light / dark cycle with free access to food and water. The rats were anesthetized with a mixture of Zoletil 50 (15 mg / kg) and xylazine hydrochloride (5 mg / kg). A heating pad was used to maintain their body temperature. α-Syn PFF (18 μg) was stereotaxically injected into the dorsal striatum (dSTR) of the left hemisphere at the coordinates: AP +0.1 mm, ML -2.5 mm, DV -4.5 mm. The first 0.5 μL was infused at a rate of 0.5 μL / min using a glass microelectrode, and then the remaining PFF was infused at a rate of 0.2 μL / min. Control animals received sterile PBS in the same manner. The rats were monitored regularly after surgical recovery.

[0946] 8.2 Preparation of the marmoset model

[0947] Marmosets (2 years old, 400 - 600 g) were housed at the Jiuting Experimental Center of the Institute of Neuroscience. Marmosets were housed two per cage under a 12-hour light / dark cycle with free access to food and water. The marmosets were anesthetized with a mixture of 0.5% isoflurane and 1% oxygen, and a heating pad was used to maintain their body temperature. α-Syn PFF (200 μg) was stereotaxically injected into the caudate nucleus and putamen of the right hemisphere. The coordinates for the caudate nucleus were: 9.5 mm interaural, 3 mm lateral, 7.6 mm depth, and the coordinates for the putamen were: 9.5 mm interaural, 6 mm lateral, 9.3 mm depth. Infusion was performed using a 10 μL microsyringe at a rate of 10 nL / sec. The marmosets were monitored regularly after postoperative recovery.

[0948] 8.3 Human subjects

[0949] Fourteen subjects were included in this experiment. All subjects or their legal guardians provided written informed consent in accordance with the Declaration of Helsinki before entering the study. This study was approved by the Ethics Review Board of Huashan Hospital Affiliated to Fudan University (HIRB) (approval number: (2024) review number (592)). This study was registered at the Chinese Clinical Trial Registry (registration number: ChiCTR2400093128).

[0950] Diagnosis was confirmed by three movement disorder specialists. The PD diagnosis was based on the Movement Disorder Society (MDS) criteria in 201563, and the MSA diagnosis was based on the MDS criteria in 202264, meeting the definition of clinically probable or clinically definite MSA. Healthy controls had no history of neurological or psychiatric diseases; in addition, they showed no deficits in neurological examinations or pre-imaging evaluations. Pre-imaging evaluations included motor and cognitive assessments, complete blood count (CBC), urine analysis (UA), liver function tests (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]), renal function tests (serum creatinine and blood urea nitrogen [BUN]), serum electrolyte analysis, and electrocardiogram (ECG). Vital signs were monitored at baseline and at 10 and 30 minutes after injection. The two-week follow-up included repeated laboratory evaluations and recording of adverse events.

[0951] 8.4. PET Imaging in Rodents and Marmosets

[0952] The above model rats and marmosets were anesthetized with 2.0 - 4.0% isoflurane during PET scanning. 18F-FD4 was injected via the tail vein at a dose of "1 - 1.5 mCi per rat" and "1.5 - 2.0 mCi for marmosets". Dynamic PET scans were performed for 90 minutes using a Siemens INVEON PET / CT scanner. CT was used for attenuation correction during PET acquisition, and the CT scan lasted approximately 5 minutes. PET images were reconstructed using a filtered back-projection algorithm with CT attenuation correction. The reconstructed pixel size was 0.2 mm × 0.2 mm × 0.8 mm, and then image analysis was performed using PMOD software. The uptake in key brain regions was analyzed using the ROI method on the Inveon platform. Each experiment for each group was performed twice.

[0953] 8.5 PET Imaging in Human Subjects

[0954] 18F-FD4 imaging was performed in 3D mode on a PET / CT scanner (u780 / UExplorer, United Imaging Healthcare, Shanghai, China) at Huashan Hospital, Fudan University, Shanghai, China. Each participant received an injection of 6 - 8 mCi of 18F-FD4. Low-dose CT transmission was acquired before the PET scan for attenuation correction. The healthy control group received dynamic imaging from 0 - 100 minutes after injection, while PD and MSA patients received 20-minute PET imaging (60 - 80 minutes after injection). Image reconstruction was performed using the ordered subset expectation maximization 3D (OSEM 3D) method. In addition, all subjects who underwent 18F-FD4 PET / CT imaging also received a separate 3D T1 MRI scan using a Siemens MAGNETOM Prisma scanner.

[0955] 8.6 Experimental results

[0956] Figure 6A The PET imaging of PFF-injected rats (right striatum, 0.5 months) developed for 50 - 60 min is shown, indicating that 18F-FD4 can effectively cross the blood-brain barrier and accumulate in the right striatum region of α-syn model rats, suggesting that this molecule can image α-syn PFF around the striatum.

[0957] Figure 6B The PET imaging of PFF-injected marmosets (right striatum, 6 months) developed for 50 - 60 min and a schematic diagram comparing the standardized uptake value ratio (SUVR) of the right striatum and frontal cortex are shown. The illustrated information shows that F18-FD4 can effectively cross the blood-brain barrier and the uptake increases in the right striatum region and frontal cortex region of α-syn model marmosets, suggesting that this molecule can image α-syn aggregation around these brain regions.

[0958] Figure 6C The PET imaging of PFF-injected rats (left striatum, 4 months) developed for 55 - 60 min is shown, indicating that FD17 (Compound of Example II-29) can effectively cross the blood-brain barrier and the uptake increases in the striatum on both sides of α-syn model rats, and the signal in the left striatum is higher than that in the right striatum. This signal is consistent with the distribution of α-syn in the model rats' brains, suggesting that FD17 can effectively label α-syn.

[0959] Figure 6D The PET imaging of PFF-injected marmosets (right striatum, 5 months) developed for 50 - 60 min is shown, indicating that FD17 can effectively cross the blood-brain barrier and accumulate in the striatum region of α-syn model marmosets, where the uptake in the right striatum is higher than that in the left striatum. Meanwhile, the uptake in the striatum of WT marmosets does not increase, suggesting that this molecule can image α-syn pathology around the striatum.

[0960] Figure 6E The PET imaging of PFF-injected rats (left striatum, 0.5 months) developed for 30 - 40 min is shown, indicating that FA35 (Compound of Example III-23) can effectively cross the blood-brain barrier and accumulate in the striatum region of α-syn model rats, suggesting that this molecule can image α-syn PFF around the striatum.

[0961] Figure 6FPET imaging of PFF-injected rats (left striatum, 0.5 months) developed for 60 - 80 min shows that FA23 (Compound I-48 of Example I) can effectively cross the blood-brain barrier and accumulate in the striatal region of α-syn model rats, indicating that this molecule can image α-syn PFF injected into the striatum.

[0962] Figure 7 PET imaging of human subjects treated with the 18F FD4 compound is shown. 18F FD4 shows a significant increase in the uptake of the radioligand, which can be observed at the midbrain level (as indicated by the arrow). The non-specific signal in the SN and SNc is very low.

[0963] Figure 8 PET imaging of human subjects treated with the prior art compound C0505 (Endo et al., Imaging α-synuclein pathologies in animal models and patients with Parkinson’s and related diseases, Neuron 112, 1 - 18, August 7, 2024) is shown. Although the total uptake of the midbrain tracer is higher in PD / DLB patients, non-specific signals are also observed in healthy volunteers (A). In addition, the SUVR values of many PD / DLB cases are close to or even lower than those of healthy volunteers (B), indicating that C0505 is ineffective for PD / DLB imaging.

[0964] All documents mentioned in the present invention are incorporated herein by reference as if each document was individually incorporated by reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

1. A compound that specifically recognizes α-synuclein aggregates, characterized in that, The compound is a compound of formula A, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein, X, Y and U are each independently selected from CH and N, and when any one of them is CH, the hydrogen atom on the CH can be substituted by R1; W is selected from CH and N; Z is selected from CH2, NH, O and S; L is selected from -CH=CH- and -C≡C-, and when n is greater than 1, -(L) n - is a chain formed by Ls that are the same as or different from each other; Ring A is selected from C 6-8 cycloalkanes, C 6-12 aryl rings, 6-10 membered heteroaryl rings and 6-8 membered heterocyclic rings; Ring B is selected from C 4-8 cycloalkanes, C 6-12 aryl rings, 5- to 10-membered heteroaryl rings, and 4- to 8-membered heterocyclic rings; M is a direct bond or a 4- to 8-membered heterocycle; Each R1 is independently selected from: deuterium, tritium, hydroxyl, amino, halogen, nitro, cyano, -COOH, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -SO2-C 1-6 alkyl, -CO-C 1-6 alkyl, -C(O)OC 1-6 alkyl, -C(O)NH2, -C(O)NHC 1-6 alkyl, -C(O)N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl, -SO2NH2, -SO2NHC 1-6 alkyl, -SO2N(C 1-6 alkyl)2, -NHSO2-C 1-6 alkyl, -N(C 1-6 alkyl)-SO2-C 1-6 alkyl, -NH-C 3-8 cycloalkyl, -N(C 1-6 alkyl)(C 3-8 cycloalkyl), -C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl, Among them, -C in R1 1-6 alkyl or C as part of a group 1-6 alkyl, -C 3-8 cycloalkyl or -C as part of a group 3-8 cycloalkyl, 3- to 8-membered heterocyclic group and 5- to 10-membered heteroaryl are each independently optionally substituted with one or more substituents, and the substituents are each independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), and two substituents attached to the same C atom optionally together with the C atom to which they are attached form a 3- to 6-membered heterocycle; R2 and R3 are each independently selected from: deuterium, tritium, hydroxyl, amino, halogen, nitro, cyano, -COOH, -C 1-6 alkyl, -C 2-6 alkenyl, -C 2-6 alkynyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -SO2-C 1-6 alkyl, -CO-C 1-6 alkyl, -C(O)OC 1-6 alkyl, -C(O)NH2, -C(O)NHC 1-6 alkyl, -C(O)N(C 1-6 alkyl)2, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl, -SO2NH2, -SO2NHC 1-6 alkyl, -SO2N(C 1-6 alkyl)2, -NHSO2-C 1-6 alkyl, -N(C 1-6 alkyl)-SO2-C 1-6 alkyl, -NH-C 3-8 cycloalkyl, -N(C 1-6 alkyl)(C 3-8 cycloalkyl), -C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl and C 6-12 aryl, wherein the -C 3-8 cycloalkyl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl and C 6-12 aryl are each independently optionally substituted by halogen, hydroxyl, -OC 1-6 alkyl, -NHC 1-6 alkyl or -N(C 1-6 alkyl)2, wherein the -C 1-6 alkyl or C 1-6 alkyl as part of another group or a substituent is optionally substituted by one or more substituents each independently selected from: deuterium, tritium, halogen, hydroxyl, -O-(3-6 membered heterocyclic group), -O-(p-toluenesulfonyl); n is an integer from 0 to 3; p is an integer from 1 to 4; q and t are each independently selected from integers from 0 to 5; wherein the heteroaryl and heterocyclic groups each independently contain 1, 2, 3 or 4 heteroatoms selected from N, S or O; and wherein each occurrence of a halogen is optionally in its isotopic form, and the carbon atoms and / or the hydrogen atoms thereon in the substituents are optionally in their isotopic forms.

2. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to claim 1, wherein W is selected from CH and N; and Z is selected from O and S.

3. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to claim 1 or 2, wherein X, Y and U are all CH, or one or two of X, Y and U are N and the rest are CH.

4. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 3, wherein the fused ring containing X, Y, U, W and Z is selected from:

5. A compound according to any one of claims 1 to 4, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is an integer from 1 to 2, and each R1 is independently selected from: halogen, nitro, hydroxy, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl and 5- to 8-membered heterocyclic groups, wherein, -C in R1 1-6 alkyl or C as part of a group 1-6 alkyl and 5- to 8-membered heterocyclic groups are each independently optionally substituted with one or more, for example 1 to 2, substituents, each independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -O-(3- to 6-membered heterocyclic group), -O-(p-toluenesulfonyl), wherein the halogen occurring in R1 is optionally in isotopic form, for example 18F.

6. A compound according to any one of claims 1 to 5, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is an integer from 1 to 2, and each R1 is independently selected from halogen, nitro, hydroxy, -O-C 1-6 alkyl, -O-C 1-6 haloalkyl, -O-C 1-6 halo-hydroxyalkyl, -O-C substituted by O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -NHC 1-6 alkyl, -NHC 1-6 haloalkyl, -NHC 1-6 halo-hydroxyalkyl, -NH-substituted by O-(3-6 membered heterocyclic group) and / or -O-(p-toluenesulfonyl) 1-6 alkyl, -N(C 1-6 alkyl)2, -COOH, -NHCO-C 1-6 alkyl, -N(C 1-6 alkyl)-CO-C 1-6 alkyl and 5-6 membered heterocyclic group, wherein the 5-6 membered heterocyclic group is optionally substituted by one or more, for example 1-2 substituents, each substituent being independently selected from deuterium, tritium, -O-C 1-6 alkyl, -C 1-6 alkyl, halogen, hydroxy, oxo, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, wherein the halogen occurring in R1 is optionally in isotopic form, such as F18.

7. A compound according to any one of claims 1 to 6, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein p is an integer from 1 to 2, and each R1 is independently selected from: F, Br, 8. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 7, wherein p is 1 and R1 is located at the ortho-ring carbon atom of Y; or p is 2 and R1 is respectively connected to Y and the ortho-ring carbon atom of Y, or R1 is respectively connected to X and the ring carbon atom between X and Y.

9. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 8, wherein p is 1.

10. A compound according to any one of claims 1 to 9, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotopic variant thereof, wherein each R1 is independently selected from 11. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 10, wherein n is not 0 and M does not exist, and formula (A) has the following sub-formula:

12. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 10, wherein n is 0 and M is a 4- to 8-membered heterocycle, and formula (A) has the following sub-formula:

13. The compound or its stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant according to any one of claims 1 to 11, wherein n is 1 and L is -CH=CH-, or L is -C≡C-; or n is 2 and (L)2 is -CH=CH-CH=CH-, -CH=CH-C≡C-, -C≡C-CH=CH- or -C≡C-C≡C-, preferably -CH=CH-CH=CH- or -CH=CH-C≡C-; or n is 3, and (L)3 is -CH=CH-CH=CH-CH=CH-, -CH=CH-CH=CH-C≡C-, -CH=CH-C≡C-CH=CH-, -C≡C-CH=CH-CH=CH-, -CH=CH-C≡C-C≡C-, -C≡C-CH=CH-C≡C- or -C≡C-C≡C-CH=CH-, preferably -CH=CH-CH=CH-CH=CH-, -CH=CH-CH=CH-C≡C-, -CH=CH-C≡C-CH=CH- or -CH=CH-C≡C-C≡C-.

16. A compound according to any one of claims 1 to 15, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is linked to the remainder of the molecule through a ring carbon atom, or when ring A is a heterocycle, it may also be linked to the remainder of the molecule through a ring heteroatom.

14. A compound according to any one of claims 1 to 13, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from C 6-10 an aromatic ring, a 6- to 8-membered heteroaromatic ring and a 6- to 8-membered heterocyclic ring, for example ring A is selected from a benzene ring and a 6-membered heteroaromatic ring or 6-membered heterocyclic ring containing 1 or 2 nitrogen heteroatoms, for example A is selected from 15. A compound according to any one of claims 1 to 14, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is selected from 23. A compound according to any one of claims 1 to 22, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein t is 1.

17. A compound according to any one of claims 1 to 16, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring A is unsubstituted or ring A is substituted by one R2, and R2 is selected from halogen, nitro, cyano and halogen-substituted -C 1-6 alkyl, such as selected from halogen, nitro and cyano, such as R2 is selected from halogen or its isotope, such as R2 is selected from F and 18F.

18. A compound according to any one of claims 1 to 17, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from C 6-10 an aromatic ring and a 5- to 8-membered heteroaromatic ring, for example ring B is selected from a benzene ring, a naphthalene ring and a 5- to 6-membered heteroaromatic ring containing 1 to 3, for example 1 or 2, heteroatoms selected from nitrogen, oxygen and sulfur, for example B is selected from 19. A compound according to any one of claims 1 to 18, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is selected from 20. A compound according to any one of claims 1 to 19, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is optionally substituted by 1 or 2 R3 groups, and R3 is selected from hydroxy, amino, halogen, nitro, cyano, -C 1-6 alkyl, -O-C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -C 3-8 cycloalkyl, 5- to 8-membered heterocyclic group and C 6-10 aryl, wherein the -C 3-8 cycloalkyl, 5- to 8-membered heterocyclic group and C 6-10 aryl are preferably -C 3-6 cycloalkyl, 5- to 7-membered heterocyclic group and C6 aryl, each independently optionally substituted by halogen, hydroxy, -OC 1-6 alkyl, -NHC 1-6 alkyl or -N(C 1-6 alkyl)2, wherein the -C 1-6 alkyl in R3 is optionally substituted by 1 or 2 substituents each independently selected from: deuterium, tritium, halogen, hydroxy, -O-(3- to 6-membered heterocyclic group) and -O-(p-toluenesulfonyl), preferably deuterium, tritium, halogen and hydroxy.

21. A compound according to any one of claims 1 to 20, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is optionally substituted by 1 or 2 R3 groups, and R3 is selected from: hydroxy, amino, F, Br, nitro, cyano, -O-CH3, -NHCH3, -N(CH3alkyl)2, 22. A compound according to any one of claims 1 to 21, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein ring B is substituted by -NHC 1-6 alkyl.

25. A compound according to any one of claims 1 to 24, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, said isotope variant comprising an isotope selected from 2H, 3H, 13C, 11C, 14C, 15N, 18O, 17O, 31P, 32P, 35S, 18F and 36Cl, preferably comprising 18F.

24. A compound according to any one of claims 1 to 8 and 10 to 17, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, wherein M is a 6- to 8-membered nitrogen-containing heterocycle, such as a 6-membered nitrogen-containing heterocycle, such as 26. A compound according to claim 1, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, selected from:

27. A compound according to claim 26, which is selected from the following compounds and their stereoisomers, or their pharmaceutically acceptable salts or solvates:

28. A compound or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof as defined in any one of claims 1 to 27 for the treatment or diagnosis of neurodegenerative diseases associated with α-synuclein aggregates and other misfolded protein aggregates.

29. A composition specifically binding to α-synuclein aggregates, comprising a compound according to any one of claims 1 to 28, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, and one or more pharmaceutically acceptable carriers.

30. Use of a compound according to any one of claims 1 to 28, or a stereoisomer, pharmaceutically acceptable salt, solvate or stable isotope variant thereof, or a composition according to claim 29 in the preparation of a medicament for the treatment or diagnosis of neurodegenerative diseases associated with α-synuclein aggregates and other misfolded protein aggregates.

31. Use according to claim 30, wherein the neurodegenerative disease is selected from Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, Alzheimer's disease, amyotrophic lateral sclerosis, progressive muscular atrophy, progressive supranuclear palsy, preferably Parkinson's disease and progressive muscular atrophy. ​ 32. A method for detecting α-synuclein aggregates in a subject, comprising the following steps: (A) Administering to the subject a safe and effective amount of a compound according to any one of claims 1 to 28 or a composition according to claim 28; and (B) Detecting the binding of the above compound or composition to α-synuclein aggregates in the subject.

33. The method according to claim 32, characterized in that, The detection in step (B) is carried out by imaging techniques, preferably by the following imaging techniques: positron emission tomography, single photon emission computed tomography, near-infrared brain functional imaging, optical imaging, or a combination thereof.

Images