CN119707781A - A compound and its preparation method and use - Google Patents

Abstract

The present invention provides a compound having a structure as shown in formula (I) or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotope-labeled substance thereof. The present invention also provides a method for preparing the compound or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotope-labeled substance thereof and use thereof in preparing a ferroptosis inhibitor.

Description

Compound and preparation method and application thereof

Technical Field

The invention relates to the field of medicines, in particular to a compound, a preparation method thereof and application thereof in preparing an iron death inhibitor.

Background

Iron death (Ferroptosis) is a programmed cell death mode driven by iron-dependent and lipid peroxidation. Iron death is morphologically characterized by a decrease in mitochondrial volume, an increase in mitochondrial membrane density, a decrease or disappearance of mitochondrial cristae, a disruption of the mitochondrial outer membrane, and a normal cell nucleus size, which is a major morphological feature of iron death that is distinguished from apoptosis, necrosis, and autophagy. The biochemical features of iron death are mainly represented by intracellular iron and ROS accumulation, activation of Mitogen Activated Protein Kinase (MAPK) signaling system, inhibition of cystine/glutamate transporter system, and increased NADPH oxidation.

More and more studies confirm that cellular iron death is closely related to many conditions, disorders and diseases. Dixon et al found, at the earliest in 2012, that this pattern of cell death was associated with small molecule induced RAS tumor cell death (Scott J Dixon et al, cell.2012 May 25;149 (5): 1060-72.Doi: 10.1016/j.cell.2012.03.042). Recently, a great deal of research has further demonstrated the role of iron death in cancer, organ damage, tissue ischemia reperfusion injury, cerebral stroke, cardiovascular disease, degenerative disorders, etc. (Bo Li et al, frontiers in Pharmacology 2020, vol.11, doi:10.3389/fphar.2020.00239; xuejun Jiang et al ,Nat Rev Mol Cell Biol.2021,vol.22(4),pp.266-282;Brent R.Stockwell,Cell.2022,vol.185,pp.2401-2421,doi.org/10.1016/j.cell.2022.06.003;Xinhua Xia et al, CELL DEATH discovery.2024, vol.10, pp.265-281, doi.org/10.1038/s 41420-024-02037-9). In particular, it relates to the occurrence and development of various cancers, neurodegenerative diseases, cardiovascular and cerebrovascular diseases, immune-related diseases, liver and kidney failure, inflammation, metabolic diseases and the like, and particularly plays an important role in diseases such as Alzheimer's disease, parkinson's disease, tumors, cerebral apoplexy, ischemia reperfusion injury, atherosclerosis, liver and kidney failure, inflammation, diabetic complications and the like. By stimulating or inhibiting the occurrence of iron death, the occurrence and progression of related diseases can be tolerated, and thus iron death inhibitors are considered potential drugs for the treatment of these diseases.

Ferrostatin-1 (Fer-1) is a known, potent, selective iron death inhibitor (Scott J Dixon et al, cell.2012 May 25;149 (5): 1060-72.Doi: 10.1016/j.cell.2012.03.042) of the formula:

however, ferrostatin-1 activity was not high, and IC50 values were higher than 100nM in various in vitro iron death cell models. Thus, how to find and prepare a more active iron death inhibitor is a challenge to be solved.

Disclosure of Invention

To remedy the deficiencies of the prior art, it is an object of the present invention to provide a compound of a completely new structure which exhibits excellent iron death suppressing activity in a plurality of cell models and thus can be used as an iron death suppressing agent for the prevention and/or treatment of various diseases.

In a first aspect the present invention provides a compound of the structure shown in formula (I) or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof,

In formula (I), ring a is selected from C ₆～C₁₂ aryl optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

Ring B is selected from a 5-12 membered heterocyclyl or a 5-12 membered heteroaryl, said heterocyclyl or heteroaryl comprising at least one N atom as a ring atom, and said heterocyclyl or heteroaryl being optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy;

Each R ₁、R₂ is independently selected from hydrogen, C ₁～C₆ alkyl, or C ₃～C₁₀ cycloalkyl;

Each R ₃ is independently selected from hydrogen, halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ haloalkoxy, C ₃～C₁₀ cycloalkyl, 5-to 12-membered heterocyclyl, C ₆～C₁₂ aryl, or 5-to 12-membered heteroaryl, said hydroxy, carboxy, amino, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl being optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

m is selected from 0, 1,2, 3 or 4.

In some embodiments according to the invention, in formula (I),

Ring a is selected from C ₆～C₁₀ aryl;

Ring B is selected from a 5-8 membered heterocyclyl group comprising 1-3N atoms as ring atoms, and which heterocyclyl group is optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy;

Each R ₁、R₂ is independently selected from hydrogen or C ₁～C₆ alkyl;

Each R ₃ is independently selected from hydrogen, halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ haloalkoxy, 5-to 8-membered heterocyclyl, or 5-to 8-membered heteroaryl, optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

m is selected from 0,1, 2 or 3.

In some preferred embodiments according to the invention, in formula (I),

Ring a is selected from phenyl;

Ring B is selected from a 5-6 membered heterocyclyl group comprising 1-2N atoms as ring atoms (which may for example be selected from piperidinyl or piperazinyl) and which heterocyclyl group is optionally substituted with one or more substituents selected from C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy or C ₁～C₄ haloalkoxy;

Each R ₁、R₂ is independently selected from hydrogen or C ₁～C₄ alkyl;

Each R ₃ is independently selected from hydrogen, F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, C ₁～C₄ haloalkoxy, or a 5-to 6-membered heterocyclyl containing at least one N atom as a ring atom (which may be selected, for example, from piperidinyl, piperazinyl, or morpholinyl), and the heterocyclyl is optionally substituted with one or more substituents selected from F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, or C ₁～C₄ haloalkoxy;

m is selected from 0, 1 or 2.

In some more preferred embodiments according to the present invention, in formula (I), R ₃ may include, but is not limited to, H, F, cl, -OCH ₃、-CF₃,

In some embodiments according to the invention, the compound has a structure as shown in formula (II):

In formula (II), each Z ₁、Z₂、Z₃ is independently selected from-C (R ₄)₂-、-NR₄ -or a bond, preferably, Z ₁、Z₂、Z₃ is not simultaneously-NR ₄ -;

Each R ₄ is independently selected from hydrogen, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy, preferably R ₄ is the same or different and is independently selected from hydrogen, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy or C ₁～C₄ haloalkoxy;

R ₁、R₂、R₃, m are each independently defined in any of the above schemes.

In some embodiments according to the invention, the compound has a structure as shown in formula (III):

In the formula (III), R ₁、R₂、R₃、R₄ and m are defined as any one of the above technical schemes independently.

In some preferred embodiments according to the present invention, in formula (III), R ₁ is selected from hydrogen, R ₂、R₄ is each independently selected from methyl, ethyl, n-propyl or isopropyl;

each R ₃ is independently selected from hydrogen, F, cl, methoxy, or trifluoromethyl;

m is selected from 0, 1 or 2.

In some embodiments according to the invention, the compound has a structure as shown in formula (IV):

in the formula (IV), Y is selected from-C (R ₆)₂-、-NR₆ -, -O-or-S-;

R ₅、R₆ is each independently selected from hydrogen, halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy, preferably R ₅、R₆ is each independently selected from hydrogen, F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, or C ₁～C₄ haloalkoxy, more preferably R ₅、R₆ is each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, carboxy, or trifluoromethyl;

n is selected from 0, 1 or 2;

R ₁、R₂、R₄ is each independently as defined in any one of the above embodiments, preferably R ₁、R₂、R₄ is each independently selected from hydrogen, methyl, ethyl, n-propyl or isopropyl, more preferably R ₁ is selected from hydrogen, and R ₂、R₄ is each independently selected from methyl, ethyl, n-propyl or isopropyl.

In some embodiments according to the invention, the compound has a structure as shown in formula (IV-1) or formula (IV-2):

In the formula (IV-1) or the formula (IV-2), Y, R ₁、R₂、R₄、R₅ and n are each independently defined in any one of the above technical schemes.

In some preferred embodiments according to the invention, in formula (IV-1) or formula (IV-2), Y is selected from-C (R ₆)₂-、-NR₆ -or-O-;

Each R ₁、R₂、R₄ is independently selected from hydrogen or C ₁～C₄ alkyl (e.g., hydrogen, methyl, ethyl, n-propyl, isopropyl, etc.);

R ₅、R₆ is each independently selected from hydrogen, carboxyl, C ₁～C₄ alkyl, or C ₁～C₄ haloalkyl (e.g., hydrogen, methyl, ethyl, n-propyl, isopropyl, carboxyl, trifluoromethyl, etc.).

In some more preferred embodiments according to the present invention, in formula (IV-1) or formula (IV-2), Y is selected from-C (R ₆)₂-、-NH-、-N(CH₃) -or-O-;

R ₁ is selected from hydrogen, R ₂、R₄ is independently selected from hydrogen, methyl, ethyl, n-propyl or isopropyl;

R ₅、R₆ is each independently selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, carboxyl or trifluoromethyl.

In some most preferred embodiments according to the present invention, the compound may include the following compounds:

The second aspect of the present invention provides a method for preparing the compound according to any one of the above technical schemes or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, wherein the preparation method comprises the steps of reacting an aniline compound with a structure shown as formula (I-1) with a halogenide with a structure shown as formula (I-2) to obtain a first intermediate with a structure shown as formula (I-3), reacting the first intermediate with p-toluenesulfonyl hydrazine to obtain a second intermediate with a structure shown as formula (I-4), and reacting the second intermediate with a boric acid compound with a structure shown as formula (I-5) to obtain a compound with a structure shown as formula (I);

wherein X is selected from halogen, for example from Br;

Ring a, ring B, R ₁、R₂、R₃, m are each independently defined as in any one of the above claims.

In some embodiments according to the present invention, the reaction materials having the structures represented by the formulas (I-1), (I-2), and (I-5) may be commercially available products or synthesized by referring to the existing literature.

In a third aspect, the present invention provides a pharmaceutical composition comprising a compound according to any one of the above technical schemes or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, and one or more pharmaceutically acceptable excipients.

A fourth aspect of the invention provides the use of a compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, or a pharmaceutical composition according to any one of the preceding claims, for the preparation of an iron death inhibitor.

The compound provided by the invention or pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof and the pharmaceutical composition provided by the invention can be used for preparing iron death inhibitors (namely medicines for inhibiting cell iron death), and can be used for preventing and/or treating various diseases, such as neurodegenerative diseases, tissue ischemia reperfusion injury, cerebral apoplexy, cardiovascular diseases, renal failure, liver injury, diabetic complications, cancers and the like by inhibiting cell iron death.

A fifth aspect of the present invention provides the use of a compound according to any one of the preceding claims, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, or a pharmaceutical composition according to any one of the preceding claims, for the manufacture of a medicament for the prevention and/or treatment of neurodegenerative diseases, tissue ischemia reperfusion injury, cerebral stroke, cardiovascular diseases, renal failure, liver injury, diabetic complications or cancer.

In some embodiments according to the invention, the neurodegenerative disease may be parkinsonism or alzheimer's disease.

In some embodiments according to the invention, the stroke may be ischemic stroke or hemorrhagic stroke.

In some embodiments according to the invention, the cancer may be pancreatic cancer.

The compounds provided by the invention or pharmaceutically acceptable salts, stereoisomers, solvates, hydrates or isotopic labels thereof, and the pharmaceutical compositions provided by the invention prevent and/or treat the diseases by inhibiting cell iron death.

The technical scheme provided by the invention has the following advantages:

(1) Compared with the existing iron death inhibitor, the compound provided by the invention has obvious chemical structure difference and has more excellent cell iron death inhibition activity, so that the compound can be used as an iron death inhibitor with great application potential, and the variety and application of the iron death inhibitor are expanded, so that the compound has very important economic and social significance.

(2) The preparation process of the compound provided by the invention is simple and convenient, the reaction condition is mild, the operation and control are convenient, and the production cost is low, so that the compound is very suitable for industrial production.

Detailed Description

Terminology

In the present invention, unless otherwise indicated, scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art.

Throughout the specification and claims, the words "comprise," "have," "include," and the like are to be construed as having an inclusive rather than an exclusive or exhaustive meaning, i.e., a meaning of "including but not limited to," unless the context clearly dictates otherwise. Unless otherwise indicated, "comprising" includes "consisting of.

The term "and/or", such as "X and/or Y", as used herein, alone or in combination, should be understood to mean "X and Y" or "X or Y" and should be used to provide explicit support for both meanings or either meaning.

As used herein, "C ₁～C_n" includes C ₁～C₂、C₁～C₃、……C₁～C_n. For example, the "C ₁～C₆" group refers to a moiety having 1 to 6 carbon atoms, i.e., the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms. Thus, for example, reference to "C ₁～C₄ alkyl" refers to an alkyl group containing 1 to 4 carbon atoms, i.e., the alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. The numerical ranges, such as "1 to 6", "6 to 12", etc., herein refer to the individual integers in the given range.

The term "alkyl" as used herein, alone or in combination, refers to an optionally substituted straight chain or optionally substituted branched saturated aliphatic hydrocarbon. The "alkyl" herein may preferably have 1 to 6 carbon atoms, for example, 1 to 6 carbon atoms, or 1 to 5 carbon atoms, or 1 to 4 carbon atoms, or 1 to 3 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-l-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-l-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-dimethyl-l-butyl, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl and the like. Where a group as defined herein, such as "alkyl" appears in numerical ranges, for example, "alkyl of C ₁～C₆" refers to an alkyl group that may be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, or 6 carbon atoms, and alkyl herein also includes instances where no numerical ranges are specified. Alkyl groups may be substituted or unsubstituted.

"Alkyl" as used herein in combination refers to an alkyl group attached to other groups, e.g., an alkyl group in an alkoxy, haloalkyl, haloalkoxy group, as defined above when used alone.

The term "haloalkyl" as used herein, alone or in combination, means that one or more, even all, of the hydrogens of the alkyl groups are replaced with halogen, and C ₁～C₆ haloalkyl includes, for example, C ₁～C₄ haloalkyl or C ₁～C₃ haloalkyl, preferably fluoroalkyl and chloroalkyl, non-limiting examples of haloalkyl include fluoromethyl, chloromethyl, difluoromethyl, dichloromethyl, trifluoromethyl, trichloromethyl, 2-fluoroethyl, 2-difluoroethyl, and the like. Haloalkyl may be substituted or unsubstituted.

The term "alkoxy", as used herein, alone or in combination, refers to an alkyl group, as defined above, attached to the parent molecular moiety through an oxygen atom, C ₁～C₆ alkoxy groups, including, for example, C ₁～C₄ alkoxy or C ₁～C₃ alkoxy groups, non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, pentyloxy, hexyloxy, and the like. Alkoxy groups may be substituted or unsubstituted.

The term "haloalkoxy" as used herein, alone or in combination, refers to replacement of one or more, or even all, of the hydrogens of the alkoxy groups with halogen, and C ₁～C₆ haloalkoxy includes, for example, C ₁～C₄ haloalkoxy or C ₁～C₃ haloalkoxy, non-limiting examples of haloalkoxy groups include monofluoromethoxy, chloromethoxy, difluoromethoxy, dichloromethoxy, trifluoromethoxy, trichloromethoxy, and the like. Haloalkoxy groups may be substituted or unsubstituted.

The term "cycloalkyl" as used herein, alone or in combination, refers to an optionally substituted non-aromatic saturated carbocyclic ring, which may include monocyclic (having one ring), bicyclic (having two rings), or polycyclic (having more than two rings), ring types including fused, bridged, and spiro rings. Preferably, the cycloalkyl group may have 3 to 10 ring-forming carbon atoms therein, for example, 3 to 8 ring-forming carbon atoms or 3 to 6 ring-forming carbon atoms. Non-limiting examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. Cycloalkyl groups may be substituted or unsubstituted.

The term "heterocyclyl", as used herein alone or in combination, refers to an optionally substituted non-aromatic 5-to 12-membered monocyclic, bicyclic or polycyclic ring of the type including fused, bridged and spiro rings, the ring forming atoms of which may contain 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Preferably, the heterocyclic group may have 5 to 10 ring-forming atoms therein, for example, 5 to 8 ring-forming atoms. Non-limiting examples of heterocyclyl groups include, but are not limited to, morpholinyl, oxetanyl, thiomorpholinyl, tetrahydrofuranyl, tetrahydropyranyl, 1-dioxo-thiomorpholinyl, piperidinyl, 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo [3.2.1] octyl, piperazinyl, hexahydropyrimidine, and the like. The heterocyclic group may be substituted or unsubstituted.

The term "aryl" as used herein, alone or in combination, refers to an optionally substituted aromatic hydrocarbon group having 6 to 12, such as 6 to 10, ring-forming carbon atoms, which may be a monocyclic aryl, bicyclic aryl, or polycyclic aryl group. The bicyclic aryl or more can be a monocyclic aryl fused to other independent rings, such as alicyclic, aromatic rings. Non-limiting examples of monocyclic aryl groups include, but are not limited to, phenyl and the like, non-limiting examples of bicyclic aryl groups include naphthyl, tetrahydronaphthyl, biphenyl and the like, and non-limiting examples of polycyclic aryl groups include, but are not limited to, phenanthryl, indenyl, anthracenyl, fluorenyl, azulenyl and the like. Aryl groups may be substituted or unsubstituted.

The term "heteroaryl" as used herein, alone or in combination, refers to an optionally substituted 5 to 12 membered monocyclic, bicyclic or polycyclic ring of the type including fused, bridged and spiro rings, the ring forming atoms of which may contain 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Preferably, the heteroaryl group may have 5 to 8 ring-forming atoms therein. Non-limiting examples of heteroaryl groups include, but are not limited to, furyl, pyridyl, 2-oxo-1, 2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2, 3-thiadiazolyl, benzodioxolyl, benzothienyl, benzimidazolyl, indolyl, isoindolyl, 1, 3-dioxo-isoindolyl, quinolinyl, indazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, isothiazolyl, 1H-1,2, 4-triazolyl, 4H-1,2, 4-triazolyl, pyridyl, pyrimidinyl, pyrazin-2 (1H) -onyl, pyrimidin-4 (3H) -onyl, pyridazin-3 (2H) -onyl, 1H-indolyl, benzo [ d ] pyrrolyl, imidazo [ d ] pyrrolyl, 3, c [ d ] pyrrolyl, 3, c ] pyrrolyl, and the like. Heteroaryl groups may be substituted or unsubstituted.

The term "halogen" as used herein, alone or in combination, refers to fluorine (F), chlorine (Cl), bromine (Br) and iodine (I).

The term "hydroxy", as used herein, alone or in combination, refers to-OH.

The term "carboxy", as used herein, alone or in combination, refers to-COOH.

The term "cyano", as used herein, alone or in combination, refers to-CN.

The term "amino" as used herein, alone or in combination, refers to-NH ₂.

The term "bond" as used herein, alone or in combination, means the absence of a substituent indicated, the two end portions of which are directly linked to form a bond.

The term "substituted" as used herein alone or in combination means that one or more hydrogen atoms, preferably 1 to 6, more preferably 1 to 3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents. The person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

The term "pharmaceutically acceptable" as used herein, alone or in combination, refers to a material that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., the material can be administered to an individual without causing an adverse biological reaction or interacting in an adverse manner with any of the components contained in the composition.

The term "pharmaceutically acceptable salt" as used herein, alone or in combination, refers to salts of the compounds of the present invention that retain the biological effectiveness and properties of the free acid or free base by reaction with a non-toxic inorganic or organic base. Available using standard procedures well known in the art. Suitable salts are listed in Remingtong's Pharmaceutical Scicences,17th ed., mack Publishing Company, easton, pa.,1985, p.1418 and Journal of Pharmaceutical Science,66,2 (1977).

The term "solvate" as used herein, alone or in combination, refers to a physical aggregate of a compound of the invention and one or more solvent molecules formed by solvation, the physical aggregate comprising varying degrees of ions and covalent bonds, such as hydrogen bonds. A "hydrate" is a solvate in which the water (H ₂ O) molecule is the solvent.

The compounds of the invention may contain asymmetric or chiral centers and thus exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, sterically hindered isomers and geometric (conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention. Unless otherwise indicated, structures described herein also include all stereoisomers, such as diastereomers, enantiomers, sterically hindered isomers and geometric (conformational) isomeric forms of such structures, e.g., R and S configurations for each asymmetric center, (Z) and (E) double bond isomers, sterically hindered isomers of biphenyl structures (see, basic organic chemistry (second edition) upper book, xing Jiyi et al ,p104-105);PAC,1996,68,2193.(Basic terminology of stereochemistry(IUPAC Recommendations 1996,on page 2201))、(Z) and (E) conformational isomers.

The carbon, hydrogen, oxygen, sulfur, nitrogen, F, cl, br, I and the like referred to in the compounds of the present invention include their isotopic conditions, the carbon, hydrogen, oxygen, sulfur or nitrogen referred to in the compounds of the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include ¹²C、¹³ C and ¹⁴ C, the isotopes of hydrogen include protium (H), deuterium (D, also known as heavy hydrogen), tritium (T, also known as super heavy hydrogen), the isotopes of oxygen include ¹⁶O、¹⁷ O and ¹⁸ O, the isotopes of sulfur include ³²S、³³S、³⁴ S and ³⁶ S, the isotopes of nitrogen include ¹⁴ N and ¹⁵ N, the isotopes of fluorine include ¹⁷ F and ¹⁹ F, the isotopes of chlorine include ³⁵ Cl and ³⁷ Cl, and the isotopes of bromine include ⁷⁹ Br and ⁸¹ Br.

The term "pharmaceutical composition" as used herein alone or in combination refers to a bioactive compound optionally admixed with at least one pharmaceutically acceptable chemical ingredient (i.e., adjuvant) including, but not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents, excipients, solvents, propellants, solubilizing agents, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure modifiers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, antioxidants, chelating agents, permeation enhancers, pH adjusting agents, buffers, plasticizers, surfactants, foaming agents, defoamers, inclusion agents, humectants, absorbents, flocculating and deflocculating agents, filter aids, release retarders, and the like.

The terms "independent" or "independently" as used herein alone or in combination mean that the definition of each group in the formula is independent of the other groups, and when two or more groups have the same selectable item, the selectable items they choose may be the same or different. For example, where the number of substituents R ₃ on the phenyl ring in formula (I) is two or more, the optional substituents for each R ₃ may be the same or different.

The terms "optional" or "optionally" used herein, alone or in combination, mean that the subsequently described event or condition may, but need not, occur, and that the description includes instances where the event or condition occurs and instances where it does not. For example, "optionally alkyl-substituted heterocyclyl" means that the alkyl group may, but need not, be present, and the description includes cases where the heterocyclyl group is substituted with an alkyl group, and cases where the heterocyclyl group is not substituted with an alkyl group.

The term "subject" or "patient" as used herein, alone or in combination, refers to an animal, including a human patient in need of treatment. In certain aspects, the disclosure may also be applied in veterinary practice to any mammal or other animal in need of such immune targeted therapies, including, but not limited to, non-human primates, dogs, felines, pigs, horses, and any other animal.

The term "treating" and other similar synonyms used herein, alone or in combination, include alleviating, alleviating or ameliorating symptoms of a disease or disorder, preventing other symptoms, ameliorating or preventing underlying metabolic causes that lead to symptoms, inhibiting the disease or disorder, e.g., arresting the development of a disease or disorder, alleviating a disease or disorder, ameliorating a disease or disorder, alleviating symptoms that result from a disease or disorder, or halting symptoms of a disease or disorder, and furthermore, the term encompasses prophylactic purposes. The term also includes obtaining a therapeutic effect and/or a prophylactic effect.

The term "room temperature" as used herein, alone or in combination, refers to 25±5 ℃.

The technical scheme of the invention is further described in detail below with reference to specific embodiments.

In the examples and comparative examples of the present invention, the raw materials or reagents used were all commercially available products, and the percentages used were all mass percentages unless otherwise specified.

In the examples and comparative examples of the present invention, silica gel column chromatography was used for purifying the compounds using 200-300 mesh silica gel as a carrier, and if not specified, the eluting system was petroleum ether/ethyl acetate.

Example 1 preparation of Compound 1

To the reaction vessel were successively added an aqueous formaldehyde solution (total 0.672g, concentration 37%,2.0 eq) and 1- (4-bromophenyl) piperazine (1 g,1.0 eq) and stirred. Formic acid (0.3831 g,2.0 eq) was added dropwise, after which the reaction was continued for 1h with a heat-preservation, at 80.+ -. 5 ℃ and TLC showed complete conversion of starting material. When the heating is turned off and the temperature of the reaction is reduced to 30 ℃, 20% NaOH aqueous solution is added into the reaction system, so that the pH value of the system is 8-9. Extraction with ethyl acetate, combining the organic phases, drying over anhydrous sodium sulfate, concentrating until no significant fraction, and transferring the material to a vacuum oven for drying at 40 ℃ gave 0.919g of compound 1b in 87% yield.

Tetrahydrofuran (10 mL) and compound 1b (900 mg,1.0 eq) were added sequentially to the reaction flask, cooled to below-60℃and n-butyllithium (1.7 mL,1.2eq, 2.5M) was added dropwise, and the mixture was stirred for 2h at the end of the addition. Trimethyl borate (550 mg,1.5eq, diluted with equal volume of tetrahydrofuran) is added dropwise, the temperature is controlled to be no more than-60 ℃ in the dropping process, after the dropping is completed, the mixture is stirred for 2 hours at a constant temperature, the mixture is naturally warmed to room temperature for 16 hours, saturated ammonium chloride solution is added dropwise to enable the pH value of the system to be about 8.3, the mixture is stirred for 2 hours, the mixture is filtered, filter cakes are sequentially leached by water and n-heptane, the filter cakes are pulped for 2 hours by a mixed solvent of water and ethyl acetate (volume ratio is 3:2), the mixture is filtered, and 528mg of compound 1C is obtained by baking in a vacuum oven at 40 ℃, and the yield is 68%.

To the reaction flask were added p-fluoronitrobenzene (2.50 g,1.0 eq), 4-methylpiperidine (1.76 g,1.0 eq) and potassium carbonate (3.67 g,1.5 eq) in this order, followed by DMF (50 mL), followed by stirring at room temperature for 24h, TLC showed complete conversion of the starting material, water (100 mL) was added to the reaction solution, stirring for 1h, filtering, washing the filter cake with water, and drying the filter cake in a vacuum oven under reduced pressure at 55℃for 16h to give 3.78g of Compound 1d in 96.85% yield.

To the reaction flask, compound 1d (3.78 g,1.0 eq) and methanol (75 mL) were added sequentially, nitrogen was replaced three times, then Pd/C catalyst (0.378 g,10% w/w) was added, and then replaced three times with hydrogen, and the reaction was stirred at room temperature under a hydrogen atmosphere for 5 hours, TLC showed complete conversion of the starting material, the reaction solution was filtered through celite pad, and the filtrate was concentrated, and the obtained crude product was purified by silica gel column chromatography to give 2.58g of compound 1e, yield: 79.02%. MS M/z (ESI): 191.1544 (M+H) ⁺.

To a reaction flask, compound 1e (0.5 g,1.0 eq), m-bromoacetophenone (0.323 g,1.0 eq), tris (dibenzylideneacetone) dipalladium (120 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (152 mg,0.1 eq), cesium carbonate (1.284 g,1.5 eq) and 1, 4-dioxane (10 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere with a heat-preservation condition at 105 ℃ for 16h, tlc showed complete conversion of the starting material, cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 250mg of compound 1f with a yield of 30.84%. MS M/z (ESI): 309.1874 (M+H) ⁺.

To the reaction flask, compound 1f (250 mg,1.0 eq), p-toluenesulfonyl hydrazide (181 mg,1.2 eq), 3M HCl in MeOH (0.14 mL,0.5 eq) and methanol (5 mL) were added in this order, and the reaction was continued under nitrogen for 6 hours at 50℃and cooled to room temperature, the reaction mixture was concentrated and purified by silica gel column chromatography to give 250mg of compound 1g in a yield of 64.75%.

1G (250 mg,1.0 eq) of the compound, 1C (174 mg,1.5 eq) of the compound, potassium carbonate (116 mg,1.6 eq), 2-methyltetrahydrofuran (3.75 mL) and 1, 4-dioxane (1.25 mL) are sequentially added into a reaction bottle, the reaction is carried out under the nitrogen environment at 80 ℃ for 18h, TLC shows complete conversion of the raw materials, the reaction liquid is cooled to room temperature, the reaction liquid is extracted by 5% sodium hydroxide aqueous solution, the organic phase is backwashed twice by water, dried by anhydrous sodium sulfate, filtered and concentrated, the silica gel column chromatography is carried out, the elution system is dichloromethane/methanol, 131mg of the compound 1 is obtained, and the yield is improved ：53.34％.MS m/z(ESI):469.3323(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.70(s,1H),7.08-7.02(m,3H),6.93(d,J＝8.4Hz,2H),6.86-6.82(m,4H),6.76(s,1H),6.71(d,J＝8.0Hz,1H),6.59(d,J＝7.6Hz,1H),3.95-3.90(m,1H),3.49(d,J＝12.0Hz,2H),3.07-3.05(m,4H),2.56(d,J＝12.0Hz,2H),2.44-2.42(m,4H),2.21(s,3H),1.68(d,J＝12.8Hz,2H),1.50-1.43(m,4H),1.29-1.22(m,2H),0.94(d,J＝6.4Hz,3H).

Example 2 preparation of Compound 2

To the reaction flask were successively added p-fluoronitrobenzene (5.0 g,1.0 eq), 4-trifluoromethylpiperidine (5.426 g,1.0 eq) and potassium carbonate (7.34 g,1.5 eq), followed by DMF (100 mL), stirred at room temperature for 24h, TLC showed complete conversion of the starting material, water (200 mL) was added to the reaction solution, stirred for 1h, filtered, water washed the filter cake, and the filter cake was dried in a vacuum oven under reduced pressure at 55℃for 16h to give 8.482g of Compound 2a in a yield of 87.28%.

To the reaction flask, compound 2a (8.0 g,1.0 eq) and methanol (160 mL) were added sequentially, nitrogen was replaced three times, then Pd/C catalyst (0.8 g,10% w/w) was added, and then replaced three times with hydrogen, and the reaction was stirred at room temperature for 5 hours under a hydrogen atmosphere, TLC showed complete conversion of the starting material, the reaction solution was filtered through celite pad, and the filtrate was concentrated, and the obtained crude product was purified by silica gel column chromatography to give 6.724g of compound 2b, yield: 94.28%. MS M/z (ESI): 245.1263 (M+H) ⁺.

To a reaction flask, compound 2b (640 mg,1.0 eq), m-bromoacetophenone (803 mg,1.0 eq), tris (dibenzylideneacetone) dipalladium (120 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (152 mg,0.1 eq), cesium carbonate (1.284 g,1.5 eq) and 1, 4-dioxane (12 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 105 ℃ for 24h, tlc showed complete conversion of the starting material, cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 350mg of compound 2C with a yield of 36.75%. MS M/z (ESI): 363.1611 (M+H) ⁺.

To the reaction flask, compound 2C (350 mg,1.0 eq), p-toluenesulfonyl hydrazide (216 mg,1.2 eq), 3M HCl in MeOH (0.16 mL,0.5 eq) and methanol (7 mL) were added in this order, the reaction was carried out under nitrogen atmosphere at 50℃for 6h, TLC showed complete conversion of the starting material, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 344mg of compound 2d in a yield of 66.84%.

To a reaction flask, compound 2d (340 mg,1.0 eq), compound 1C (212 mg,1.5 eq), potassium carbonate (142 mg,1.6 eq), 2-methyltetrahydrofuran (5.1 mL) and 1, 4-dioxane (1.7 mL) were sequentially added, the reaction was allowed to stand for 24 hours under nitrogen atmosphere at 80 ℃, the reaction solution was cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with an elution system of dichloromethane/methanol to obtain 148mg of compound 2 in yield ：44.17％.MS m/z(ESI):523.2964(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.76(s,1H),7.24(d,J＝8.0Hz,2H),7.06-7.02(m,2H),6.94(d,J＝8.8Hz,2H),6.85(t,J＝9.2Hz,4H),6.74(d,J＝7.6Hz,2H),5.40(s,1H),3.61(d,J＝12.0Hz,2H),3.09-3.07(m,4H),2.62(t,J＝12.0Hz,2H),2.45-2.43(m,5H),2.21(s,3H),1.88(d,J＝12.8Hz,2H),1.75(s,3H),1.63-1.53(m,2H).

To the reaction flask, p-fluoronitrobenzene (2.5 g,1.0 eq), morpholine (1.544 g,1.0 eq) and potassium carbonate (3.674 g,1.5 eq) were added in this order, DMF (50 mL) was added, the reaction was stirred at room temperature for 5h, TLC showed complete conversion of the starting material, ice water (150 mL) was added to the reaction solution, stirring was carried out for 1h, filtration was carried out, the filter cake was washed with water, and the filter cake was dried in a vacuum oven under reduced pressure at 55℃for 16h to give 3.56g of Compound 3a in 96.49% yield.

To the reaction flask, compound 3a (3.51 g,1.0 eq) and methanol (70 mL) were added sequentially, nitrogen was replaced three times, then Pd/C catalyst (0.35 g,10% w/w) was added, and then replaced three times with hydrogen, and the reaction was stirred at room temperature for 5h under a hydrogen atmosphere, TLC showed complete conversion of the starting material, the reaction solution was filtered through celite, and the filtrate was concentrated to give 3.0g of compound 3b, which was directly used in the next step, yield: 99.85%.

To a reaction flask, compound 3b (1.864 g,1.0 eq), m-bromoacetophenone (2.5 g,1.2 eq), tris (dibenzylideneacetone) dipalladium (480 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (610 mg,0.1 eq), cesium carbonate (5.11 g,1.5 eq) and 1, 4-dioxane (30 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere with a heat-preservation condition at 105 ℃ for 15h, tlc showed complete conversion of the starting material, cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 833mg of compound 3C in a yield of 26.88%.

To the reaction flask, compound 3C (830 mg,1.0 eq), p-toluenesulfonyl hydrazine (630 mg,1.2 eq), 3M HCl in MeOH (0.47 mL,0.5 eq) and methanol (16 mL) were added in this order, the reaction was carried out under nitrogen for 6h at 50℃with TLC showing complete conversion of the starting material, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 505mg of compound 3d in a yield of 38.81%. MS M/z (ESI): 465.1887 (M+H) ⁺.

Sequentially adding a compound 3d (500 mg,1.0 eq), a compound 1C (360 mg,1.5 eq), potassium carbonate (240 mg,1.6 eq), 2-methyltetrahydrofuran (7.5 mL) and 1, 4-dioxane (2.5 mL) into a reaction bottle, carrying out heat preservation reaction for 16h at 80 ℃ under nitrogen environment, cooling to room temperature, extracting a reaction solution with a 5% sodium hydroxide aqueous solution, backwashing an organic phase with water twice, drying with anhydrous sodium sulfate, filtering and concentrating, purifying by silica gel column chromatography, eluting with dichloromethane/methanol to obtain 256mg of the compound 3, and obtaining the yield ：52.09％.MS m/z(ESI):457.2961(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.78(s,1H),7.07(t,J＝8.0Hz,3H),6.99(d,J＝8.4Hz,2H),6.86-6.83(m,4H),6.78–6.72(m,2H),6.61(d,J＝7.6Hz,1H),3.93(q,J＝7.2Hz,1H),3.71(s,3H),3.37(s,4H),3.06(t,J＝4.8Hz,4H),2.45-2.42(m,4H),2.22-2.20(m,4H),1.49(d,J＝7.2Hz,3H).

EXAMPLE 4 preparation of Compound 4

M-fluoronitrobenzene (2.5 g,1.0 eq), 4-methylpiperidine (1.76 g,1.0 eq) and potassium carbonate (3.674 g,1.5 eq) were added sequentially to a reaction flask, DMF (50 mL) was added, the mixture was stirred at 110℃for 50h, cooled to room temperature, water and ethyl acetate were added to the reaction mixture, extraction was performed, the separated liquid was backwashed twice with saturated aqueous NaCl solution, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography to give 2.74g of compound 4a in a yield of 70.2%.

To the reaction flask, compound 4a (2.74 g,1.0 eq) and methanol (30 mL) were added sequentially, nitrogen was replaced three times, then Pd/C catalyst (0.274 g,10% w/w) was added, and then replaced three times with hydrogen, and then the reaction was stirred at room temperature under a hydrogen atmosphere for 6 hours, TLC showed complete conversion of the starting material, the reaction solution was filtered through celite pad, the filtrate was concentrated, and purified by silica gel column chromatography to give 1.32g of compound 4b in 55.7% yield.

To a reaction flask, compound 4b (1.219 g,1.0 eq), m-bromoacetophenone (1.534 g,1.2 eq), palladium acetate (72 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (402 mg,0.1 eq), cesium carbonate (3.13 g,1.5 eq) and toluene (40 mL) were added in this order, and the reaction was allowed to stand for 24 hours at 110℃under nitrogen atmosphere, cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated and purified by silica gel column chromatography to give 652mg of compound 4C in 33.0% yield.

To the reaction flask was added compound 4C (719 mg,1.0 eq), p-toluenesulfonyl hydrazine (521 mg,1.2 eq), 3M HCl in MeOH (0.4 mL,0.5 eq) and methanol (15 mL) in this order, and the reaction was incubated at 50℃for 12h under nitrogen, TLC showed substantially complete conversion of the starting material, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 384mg of compound 4d in 34.56% yield.

Sequentially adding a compound 4d (384 mg,1.0 eq), a compound 1C (266 mg,1.5 eq), potassium carbonate (178 mg,1.6 eq), 2-methyltetrahydrofuran (6 mL) and1, 4-dioxane (2 mL) into a reaction bottle, carrying out heat preservation reaction for 12h at 80 ℃ under the nitrogen environment, cooling to room temperature, extracting the reaction solution by using a 5% sodium hydroxide aqueous solution, backwashing an organic phase by using water twice, drying by using anhydrous sodium sulfate, filtering and concentrating, purifying by using a silica gel column chromatography, wherein an elution system is dichloromethane/methanol, and obtaining 378mg of the compound 4, wherein the yield is obtained ：31.22％.MS m/z(ESI):469.3319(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.94(s,1H),7.13–7.07(m,3H),7.01(t,J＝8.0Hz,1H),6.92(s,1H),6.84(d,J＝8.0Hz,3H),6.69(d,J＝8.0Hz,1H),6.56(s,1H),6.44-6.38(m,2H),3.95(q,J＝7.2Hz,1H),3.52(d,J＝12.4Hz,2H),3.07-3.05(m,4H),2.58(t,J＝12.0Hz,2H),2.45-2.42(t,J＝4.9Hz,4H),2.21(s,3H),1.64(d,J＝13.2Hz,2H),1.50(d,J＝7.2Hz,4H),1.26–1.17(m,3H),0.92(d,J＝6.4Hz,2H).

EXAMPLE 5 preparation of Compound 5

Aniline (1.064 g,1.0 eq), m-bromoacetophenone (2.5 g,1.2 eq), palladium acetate (128 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (711 mg,0.1 eq), cesium carbonate (7.44 g,1.5 eq) and toluene (35 mL) were added to the reaction flask in this order, the reaction was carried out under nitrogen atmosphere at 110 ℃ for 12h, cooled to room temperature, the reaction solution was filtered with celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 1.02g of compound 6a with a yield of 42.29%. MS M/z (ESI): 212.1069 (M+H) ⁺.

To the reaction flask was added compound 5a (1.02 g,1.0 eq), p-toluenesulfonyl hydrazine (1.079 g,1.2 eq), 3M HCl in MeOH (0.8 mL,0.5 eq) and methanol (20.5 mL) in this order, and the reaction was continued under nitrogen for 4h at 50℃until the starting material was substantially converted, cooled to room temperature, the reaction mixture was concentrated, and purified by silica gel column chromatography to give 1.76g of compound 5b in a yield of 96.02%.

Sequentially adding a compound 5b (1.0 g,1.0 eq), a compound 1C (0.87 g,1.5 eq), potassium carbonate (583 mg,1.6 eq), 2-methyltetrahydrofuran (15 mL) and 1, 4-dioxane (5 mL) into a reaction bottle, carrying out heat preservation reaction for 16h at 80 ℃ under nitrogen environment, cooling to room temperature, extracting a reaction solution by using a 5% sodium hydroxide aqueous solution, backwashing an organic phase by using water for two times, drying by using anhydrous sodium sulfate, filtering and concentrating, purifying by using silica gel column chromatography, eluting with methylene dichloride/methanol to obtain 561mg of the compound 5, and obtaining the yield ：31.22％.MS m/z(ESI):372.2437(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝8.10(s,1H),7.20(t,J＝7.6Hz,2H),7.14-7.08(m,3H),7.02(d,J＝8.0Hz,2H),6.90–6.84(m,4H),6.79(t,J＝7.3Hz,1H),6.72(d,J＝7.2Hz,1H),3.97(q,J＝7.2Hz,1H),3.06(t,J＝5.2Hz,4H),2.43(t,J＝5.2Hz,4H),2.21(s,3H),1.51(d,J＝7.2Hz,3H).

EXAMPLE 6 preparation of Compound 6

To the reaction flask were successively added p-fluoroaniline (1.84 g,1.0 eq), m-bromoacetophenone (2.5 g,1.2 eq), palladium acetate (128 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (711 mg,0.1 eq), cesium carbonate (7.44 g,1.5 eq) and toluene (35 mL), reacted under nitrogen atmosphere at 110 ℃ for 12 hours, cooled to room temperature, the reaction solution was filtered with celite, and the filtrate was concentrated to obtain 1.37g of compound 6a by silica gel column chromatography, yield: 42.96%. MS M/z (ESI): 280.0943 (M+H) ⁺.

To the reaction flask, compound 6a (0.7 g,1.0 eq), p-toluenesulfonyl hydrazine (560 mg,1.2 eq), 3M HCl in MeOH (0.43 mL,0.5 eq) and methanol (14 mL) were added in this order, the reaction was carried out under nitrogen for 6h at 50℃until the conversion of the starting material was substantially complete, cooled to room temperature, the reaction mixture was concentrated and purified by silica gel column chromatography to give 0.706g of compound 6b in 62.92% yield.

Sequentially adding a compound 6b (600 mg,1.0 eq), a compound 1C (447 mg,1.5 eq), potassium carbonate (287 mg,1.6 eq), 2-methyltetrahydrofuran (9 mL) and 1, 4-dioxane (3 mL) into a reaction bottle, carrying out heat preservation reaction for 16h at 80 ℃ under nitrogen environment, cooling to room temperature, extracting the reaction solution with a 5% sodium hydroxide aqueous solution, backwashing an organic phase with water twice, drying with anhydrous sodium sulfate, filtering and concentrating, purifying by silica gel column chromatography, wherein an elution system is dichloromethane/methanol, obtaining 320mg of the compound 6 with the yield ：56％.MS m/z(ESI):440.2307(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝8.65(s,1H),7.48(d,J＝8.4Hz,2H),7.21(t,J＝8.0Hz,1H),7.09(t,J＝8.8Hz,4H),7.00–6.96(m,2H),6.88-6.85(m,3H),4.06-3.99(m,1H),3.07(t,J＝4.8Hz,4H),2.44(t,J＝4.8Hz,4H),2.22(s,3H),1.53(d,J＝7.2Hz,3H).

EXAMPLE 7 preparation of Compound 7

To the reaction flask, p-fluoronitrobenzene (2.5 g,1.0 eq), N-methylpiperazine (1.775 g,1.0 eq) and potassium carbonate (3.674 g,1.5 eq) were added in this order, DMF (50 mL) was added, stirring was performed for 2h at 90℃and TLC showed complete conversion of the starting material, ice water (150 mL) was added to the reaction solution, stirring was performed for 1h, filtering, washing the filter cake with water, and placing the filter cake in a reduced pressure vacuum oven, drying at 55℃for 16h to give 3.065g of Compound 7a, yield: 78.19%.

To the reaction flask, compound 7a (2.76 g,1.0 eq) and methanol (55 mL) were added sequentially, nitrogen was replaced three times, then Pd/C catalyst (0.275 g,10% w/w) was added, and then replaced three times with hydrogen, and the reaction was stirred at room temperature under a hydrogen atmosphere for 5 hours, TLC showed complete conversion of the starting material, the reaction solution was filtered through celite pad, and the filtrate was concentrated to give 2.3g of compound 7b, which was directly used in the next step, yield: 96.39%.

To a reaction flask, compound 7b (1.845 g,1.0 eq), m-bromoacetophenone (2.3 g,1.2 eq), palladium acetate (110 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (600 mg,0.1 eq), cesium carbonate (4.72 g,1.5 eq) and toluene (15 mL) were added in this order, the reaction was carried out under nitrogen atmosphere at 110 ℃ for 16h, cooled to room temperature, the reaction solution was filtered with celite, and the filtrate was concentrated to obtain 1.33g of compound 7C by silica gel column chromatography, the yield was 44.63%.

To the reaction flask was added compound 7C (1.33 g,1.0 eq), p-toluenesulfonyl hydrazine (0.969 g,1.2 eq), 3M HCl in MeOH (0.72 mL,0.5 eq) and methanol (27 mL) in this order, and the reaction was continued under nitrogen for 16h at 50℃until the starting material was substantially converted, cooled to room temperature, the reaction mixture was concentrated, and purified by silica gel column chromatography to give 1.607g of compound 7d in 78.21% yield.

To a reaction flask, compound 7d (1.0 g,1.0 eq), compound 1C (0.69 g,1.5 eq), potassium carbonate (0.46 g,1.6 eq), 2-methyltetrahydrofuran (15 mL) and 1, 4-dioxane (6 mL) were added in this order, the reaction was allowed to stand under nitrogen for 16 hours at 80℃and cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with methylene chloride/methanol as an eluting system to give 345mg of compound 7 in a yield ：35.08％.MS m/z(ESI):470.3248(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.72(s,1H),7.08–7.02(m,3H),6.94(d,J＝8.8Hz,2H),6.86–6.83(m,4H),6.76-6.75(m,1H),6.73-6.70(m,1H),6.59(d,J＝7.2Hz,1H),3.92(q,J＝7.2Hz,1H),3.08-3.02(m,7H),2.46-2.42(m,9H),2.22(d,J＝4.4Hz,6H),1.49(d,J＝7.2Hz,3H).

EXAMPLE 8 preparation of Compound 8

Methyl 1-t-butoxycarbonyl-4-methylpiperidine-4-carboxylate (1.0 g,1.0 eq) and ethyl acetate (20 mL) were added sequentially to the reaction flask, and after stirring to dissolve, p-toluenesulfonic acid monohydrate (2.22 g,3.0 eq) was added, and the reaction mixture was stirred at 65℃for 0.5h, and concentrated to dryness to give compound 8a, which was directly used in the next step.

To the reaction flask, the unpurified compound 8a obtained in the previous step, DMF (25 mL), p-fluoronitrobenzene (550 mg,1.0 eq) and potassium carbonate (2.69 g,5 eq) were added in this order, the reaction was stirred at room temperature for 24 hours, water (75 mL) was added, stirring was continued for 0.5 hour, filtration, washing of the filter cake with water, and drying under reduced pressure at 50℃gave 988mg of compound 8b, yield: 91.36%.

To the reaction flask, compound 8b (988 mg,1.0 eq) and methanol (15 mL) were successively added, nitrogen was substituted three times, then Pd/C catalyst (99 mg,10% w/w) was added, and then the mixture was replaced three times with hydrogen, and the reaction was stirred at room temperature for 2 hours under a hydrogen atmosphere, and the reaction solution was filtered through celite, and the filtrate was concentrated to give 817mg of compound 8C in a yield of 92.84%.

To a reaction flask, compound 8C (817 mg,1.0 eq), m-bromoacetophenone (786 mg,1.2 eq), palladium acetate (37 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (205 mg,0.1 eq), cesium carbonate (1.61 g,1.5 eq) and toluene (15 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 110 ℃ for 16 hours, cooled to room temperature, and the reaction solution was filtered with celite pad, and the filtrate was concentrated and purified by silica gel column chromatography to obtain 695mg of compound 8d with a yield of 57.64%.

To the reaction flask, compound 8d (695 mg,1.0 eq), p-toluenesulfonyl hydrazine (425 mg,1.2 eq), 3M HCl in MeOH (0.32 mL,0.5 eq) and methanol (14 mL) were added in this order, and the reaction was carried out under nitrogen for 6 hours at 45℃and cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to obtain 707mg of compound 8e in a yield of 69.72%.

To the reaction flask, compound 8e (700 mg,1.0 eq), compound 1C (430 mg,1.5 eq), potassium carbonate (290 mg,1.6 eq), 2-methyltetrahydrofuran (10.5 mL) and 1, 4-dioxane (3.5 mL) were added in this order, the reaction was allowed to stand for 16 hours at 80℃under nitrogen atmosphere, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and the silica gel column was purified by chromatography with methylene chloride/methanol as an elution system to obtain 462mg of compound 8f in 67% yield.

Sequentially adding a compound 8f (349mg, 1.0 eq), tetrahydrofuran (4 mL), methanol (0.7 mL) and water (1 mL) into a reaction bottle, uniformly dispersing, adding lithium hydroxide monohydrate (82 mg,3.0 eq), stirring at 50 ℃ for reacting for 5 hours, concentrating to dryness, adding water-dispersible agent, adjusting pH=6-7 by using saturated citric acid aqueous solution, extracting and separating liquid by using dichloromethane, merging organic phases, backwashing the organic phases by using saturated sodium chloride solution, drying by using anhydrous sodium sulfate, filtering, concentrating, purifying by using silica gel column chromatography, obtaining 198mg of compound 8 by using a eluting system of dichloromethane/methanol, and obtaining the yield ：59.48％.MS m/z(ESI):513.3230(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.72(s,1H),7.08-7.02(m,3H),6.92-6.81(m,6H),6.73-6.69(m,2H),6.60(d,J＝7.6Hz,1H),3.95-3.90(m,1H),3.27-3.22(m,2H),3.08-3.06(m,4H),2.76(t,J＝10.8Hz,2H),2.48-2.45(m,4H),2.23(s,3H),2.06(d,J＝13.2Hz,2H),1.49(d,J＝6.8Hz,5H),1.16(s,3H).

EXAMPLE 9 preparation of Compound 9

To the reaction flask, p-fluoronitrobenzene (2.5 g,1.0 eq), 2, 6-dimethylmorpholine (2.041 g,1.0 eq) and potassium carbonate (3.6754 g,1.5 eq) were added in this order, DMF (50 mL) was added, the reaction was stirred at 90℃for 4 hours, ice water (150 mL) was added to the reaction mixture, stirred for 1 hour, filtered, the filter cake was washed with water, and the filter cake was dried at 55℃for 16 hours in a vacuum oven under reduced pressure to give 4.006g of Compound 9a in 96.49% yield.

To the reaction flask, compound 9a (3.95 g,1.0 eq) and methanol (80 mL) were sequentially added, nitrogen was replaced three times, pd/C catalyst (0.390 g,10% w/w) was further added, and replaced three times with hydrogen, the reaction was stirred at room temperature under a hydrogen atmosphere for 4 hours, the reaction solution was filtered through celite, and the filtrate was concentrated to obtain 3.57g of compound 9b, which was directly used in the next step.

To a reaction flask, compound 9b (1.193 g,1.0 eq), m-bromoacetophenone (1.382 g,1.2 eq), palladium acetate (65 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (376 mg,0.1 eq), cesium carbonate (2.836 g,1.5 eq) and toluene (21 mL) were added in this order, and the reaction was allowed to stand for 16h at 110℃under nitrogen atmosphere, cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated and purified by silica gel column chromatography to give 729mg of compound 9C in a yield of 38.85%.

To the reaction flask, compound 9C (729 mg,1.0 eq), p-toluenesulfonyl hydrazine (502 mg,1.2 eq), 3M HCl in MeOH (0.4 mL,0.5 eq) and methanol (15 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 50℃for 6 hours, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to obtain 615mg of compound 9d in a yield of 55.56%.

Sequentially adding a compound 9d (616 mg,1.0 eq), a compound 1C (412 mg,1.5 eq), potassium carbonate (276 mg,1.6 eq), 2-methyltetrahydrofuran (10 mL) and 1, 4-dioxane (3 mL) into a reaction bottle, carrying out heat preservation reaction for 16h at 80 ℃ under the nitrogen environment, cooling to room temperature, extracting the reaction solution by using a 5% sodium hydroxide aqueous solution, backwashing an organic phase by using water twice, drying by using anhydrous sodium sulfate, filtering and concentrating, purifying by using a silica gel column chromatography, wherein an elution system is dichloromethane/methanol, and obtaining 77mg of the compound 9, wherein the yield is ：12.73％.MS m/z(ESI):485.3277(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.73(s,1H),7.20(t,J＝7.6Hz,1H),7.08–7.02(m,3H),6.93(t,J＝9.6Hz,3H),6.84(d,J＝8.4Hz,3H),6.79–6.71(m,2H),6.59(d,J＝7.6Hz,1H),3.73-3.65(m,2H),3.11(t,J＝4.8Hz,2H),3.06(t,J＝4.8Hz,3H),2.46-2.42(m,6H),2.22–2.15(m,6H),1.49(d,J＝7.2Hz,3H),1.14(d,J＝6.0Hz,4H).

Example 10 preparation of Compound 10

To the reaction flask, 2-fluoro-4-chloro-aniline (2.0 g,1.0 eq), m-bromoacetophenone (3.271 g,1.2 eq), palladium acetate (154 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (890 mg,0.1 eq), cesium carbonate (6.710 g,1.5 eq) and toluene (50 mL) were added in this order, and the reaction was allowed to proceed under nitrogen atmosphere at 110℃for 24 hours, cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated and purified by silica gel column chromatography to give 985mg of compound 10a in 27.19% yield.

To the reaction flask, compound 10a (1.051 g,1.0 eq), p-toluenesulfonyl hydrazine (891 mg,1.2 eq), 3M HCl in MeOH (0.67 mL,0.5 eq) and methanol (20 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 50℃for 6 hours, cooled to room temperature, the reaction mixture was concentrated, and purified by silica gel column chromatography to give 1.409g of compound 10b in a yield of 81.82%.

To a reaction flask, compound 10b (1.409 g,1.0 eq), compound 1C (1.077 g,1.5 eq), potassium carbonate (721 mg,1.6 eq), 2-methyltetrahydrofuran (21 mL) and 1, 4-dioxane (7 mL) were added in this order, the reaction was allowed to stand for 12 hours under nitrogen atmosphere at 80℃and cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed with water twice, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with an elution system of dichloromethane/methanol to give 812mg of compound 10 in yield ：58.71％.MS m/z(ESI):424.1947(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.99(s,1H),7.38-7.35(m,1H),7.22–7.07(m,5H),6.89–6.76(m,5H),3.97(q,J＝7.2Hz,1H),3.08-3.05(m,4H),2.43(t,J＝4.8Hz,4H),2.21(s,3H),1.51(d,J＝7.2Hz,3H).

EXAMPLE 11 preparation of Compound 11

To a reaction flask, p-methoxyaniline (1.69 g,1.0 eq), m-bromoacetophenone (3.28 g,1.2 eq), tris (dibenzylideneacetone) dipalladium (630 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (800 mg,0.1 eq), cesium carbonate (6.72 g,1.5 eq) and 1, 4-dioxane (40 mL) were added in this order, and the reaction was incubated under nitrogen for 12h at 105℃until the starting materials were completely converted, cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 1.93g of compound 11a in a yield of 58.29%.

To the reaction flask, compound 11a (1.93 g,1.0 eq), p-toluenesulfonyl hydrazine (1.79 g,1.2 eq), 3M HCl in MeOH (1.33 mL,0.5 eq) and methanol (38 mL) were added in this order, and the reaction was continued under nitrogen for 6 hours at 50℃and cooled to room temperature, the reaction mixture was concentrated and purified by silica gel column chromatography to give 1.0g of compound 11b in a yield of 30.53%.

To a reaction flask, compound 11b (1.0 g,1.0 eq), compound 1C (0.81 g,1.5 eq), potassium carbonate (540 mg,1.6 eq), 2-methyltetrahydrofuran (15 mL) and 1, 4-dioxane (5 mL) were sequentially added, the reaction was allowed to stand at 80℃for 16 hours under nitrogen atmosphere, the reaction mixture was cooled to room temperature, the reaction mixture was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and the silica gel column was purified by chromatography with methylene chloride/methanol as an eluting system to obtain 420mg of compound 11 in a yield 42.83％.MS m/z(ESI):402.2539(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.78(s,1H),7.08–6.98(m,5H),6.85–6.82(m,4H),6.77(s,1H),6.73(d,J＝8.4Hz,1H),6.60(d,J＝7.6Hz,1H),3.93(q,J＝7.2Hz,1H),3.70(s,3H),3.07-3.45(m,4H),2.45-2.41(m,4H),2.20(s,3H),1.49(d,J＝7.2Hz,3H).

EXAMPLE 12 preparation of Compound 12

To a reaction flask, m-methoxyaniline (1.0 g,1.0 eq), m-bromoacetophenone (1.94 g,1.2 eq), palladium acetate (91 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (506 mg,0.1 eq), cesium carbonate (3.97 g,1.5 eq) and toluene (20 mL) were added in this order, and the reaction was allowed to stand for 12 hours at 110℃under nitrogen atmosphere, cooled to room temperature, the reaction solution was filtered through celite, and the filtrate was concentrated and purified by silica gel column chromatography to give 1.092g of compound 12a in a yield of 55.74%.

To the reaction flask, compound 12a (1.092 g,1.0 eq), p-toluenesulfonyl hydrazine (1.01 g,1.2 eq), 3M HCl in MeOH (0.8 mL,0.5 eq) and methanol (20 mL) were added in this order, and the reaction was continued under nitrogen for 5 hours at 50℃and cooled to room temperature, the reaction mixture was concentrated and purified by silica gel column chromatography to give 1.273g of compound 12b in a yield of 68.7%.

To a reaction flask, compound 12b (1.2793 g,1.0 eq), compound 1C (1.03 g,1.5 eq), potassium carbonate (690 mg,1.6 eq), 2-methyltetrahydrofuran (15 mL) and 1, 4-dioxane (5 mL) were added in this order, the reaction was allowed to stand for 12 hours under nitrogen atmosphere at 80℃and cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with an elution system of methylene chloride/methanol to obtain 626mg of compound 12 in yield ：50.15％.MS m/z(ESI):402.2553(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝8.11(s,1H),7.15–7.07(m,4H),6.93–6.88(m,2H),6.85(d,J＝8.4Hz,2H),6.74(d,J＝7.6Hz,1H),6.60–6.55(m,2H),6.36(dd,J＝8.4,2.4Hz,1H),3.98(q,J＝7.2Hz,1H),3.66(s,3H),3.06(t,J＝4.8Hz,4H),2.44–2.42(m,4H),2.21(s,3H),1.51(d,J＝7.2Hz,3H).

EXAMPLE 13 preparation of Compound 13

To the reaction flask were successively added the compounds o-methoxyaniline (1.0 g,1.0 eq), m-bromoacetophenone (2.1 g,1.3 eq), tris (dibenzylideneacetone) dipalladium (370 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (470 mg,0.1 eq), cesium carbonate (3.97 g,1.5 eq) and 1, 4-dioxane (20 mL), and the reaction was carried out under nitrogen atmosphere with a heat-preservation condition at 105 ℃ for 12h, tlc showed complete conversion of the starting material, cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 0.924g of compound 13a in a yield of 47.17%.

To the reaction flask, compound 13a (924 mg,1.0 eq), p-toluenesulfonyl hydrazine (850 mg,1.2 eq), 3M HCl in MeOH (0.7 mL,0.5 eq) and methanol (20 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 50℃for 5 hours, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 0.719g of compound 13b in a yield of 45.85%.

To a reaction flask, compound 13b (719 mg,1.0 eq), compound 1C (580 mg,1.5 eq), potassium carbonate (390 mg,1.6 eq), 2-methyltetrahydrofuran (10.8 mL) and 1, 4-dioxane (3.6 mL) were sequentially added, the reaction was allowed to stand for 12 hours under nitrogen atmosphere at 80℃and cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed with water twice, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with an elution system of methylene chloride/methanol to give 543mg of compound 13 in yield ：77.02％.MS m/z(ESI):402.2542(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.23(s,1H),7.14(dd,J＝7.6,2.0Hz,1H),7.10-7.06(m,3H),6.98(dd,J＝7.6,2.0Hz,1H),6.93-6.91(m,1H),6.88-6.79(m,5H),6.68(d,J＝7.6Hz,1H),3.95(q,J＝7.2Hz,1H),3.79(s,3H),3.06(t,J＝6.0Hz,4H),2.44–2.42(m,4H),2.21(s,3H),1.50(d,J＝7.2Hz,3H).

EXAMPLE 14 preparation of Compound 14

To the reaction flask, 2, 4-dimethoxyaniline (1.245 g,1.0 eq), m-bromoacetophenone (2.1 g,1.3 eq), tris (dibenzylideneacetone) dipalladium (370 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (470 mg,0.1 eq), cesium carbonate (3.97 g,1.5 eq) and 1, 4-dioxane (25 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere with a heat-preservation condition of 105 ℃ for 12h, tlc showed complete conversion of the raw materials, and was cooled to room temperature, the reaction solution was filtered with celite, and the filtrate was concentrated and purified by silica gel column chromatography to give 400mg of compound 14a in 18.16% yield.

To the reaction flask, 14a (400 mg,1.0 eq), p-toluenesulfonyl hydrazide (399 mg,1.2 eq), 3M HCl in MeOH (0.3 mL,0.5 eq) and methanol (8 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 50℃for 5 hours, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 579mg of 14b, yield: 89.35%.

To a reaction flask, compound 14b (579 mg,1.0 eq), compound 1C (440 mg,1.5 eq), potassium carbonate (290 mg,1.6 eq), 2-methyltetrahydrofuran (8.7 mL) and 1, 4-dioxane (2.9 mL) were sequentially added, the reaction was allowed to stand for 12 hours at 80℃under nitrogen atmosphere, the reaction solution was cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed twice with water, dried over anhydrous sodium sulfate, filtered and concentrated, and the silica gel column was purified by chromatography with methylene chloride/methanol as an eluting system to obtain 364mg of compound 14 in yield ：64.03％.MS m/z(ESI):432.2657(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.08-7.04(m,3H),7.01–6.97(m,2H),6.84(d,J＝8.8Hz,2H),6.70(t,J＝2.0Hz,1H),6.61(d,J＝2.8Hz,1H),6.57–6.53(m,2H),6.45(dd,J＝8.8,2.8Hz,1H),3.89(q,J＝7.2Hz,1H),3.74(s,6H),3.06(t,J＝4.8Hz,4H),2.43(t,J＝4.8Hz,4H),2.21(s,3H),1.48(d,J＝7.2Hz,3H).

EXAMPLE 15 preparation of Compound 15

M-fluoronitrobenzene (2.0 g,1.0 eq), 4-trifluoromethyl piperidine (2.17 g,1.0 eq) and potassium carbonate (2.94 g,1.5 eq) are added into a reaction bottle in sequence, DMSO (30 mL) is added, stirring reaction is carried out for 50h at 120 ℃, cooling is carried out to room temperature, water and ethyl acetate are added into the reaction liquid, extraction and liquid separation are carried out, the extraction liquid is backwashed twice by saturated NaCl aqueous solution, anhydrous sodium sulfate is dried, filtration and concentration are carried out, and silica gel column chromatography purification are carried out, thus obtaining 1.275g of compound 15a, and the yield is 32.8%.

To the reaction flask, compound 15a (1.275 g,1.0 eq) and methanol (30 mL) were added successively, nitrogen was replaced three times, pd/C catalyst (0.127 g,10% w/w) was added three times, and then the mixture was replaced three times with hydrogen, the reaction was stirred at room temperature under a hydrogen atmosphere for 2 hours, the reaction solution was filtered through celite, and the filtrate was concentrated to obtain 1.071g of compound 15b, yield: 94.31%.

To a reaction flask, compound 15b (1.0 g,1.0 eq), m-bromoacetophenone (1.22 g,1.5 eq), palladium acetate (46 mg,0.05 eq), 1 '-binaphthyl-2, 2' -bisdiphenylphosphine (250 mg,0.1 eq), cesium carbonate (2.0 g,1.5 eq) and toluene (15 mL) were added in this order, and the reaction was allowed to stand for 12 hours at 110 ℃ under nitrogen atmosphere, cooled to room temperature, the reaction solution was filtered with celite, and the filtrate was concentrated to obtain 547mg of compound 15C by silica gel column chromatography, with a yield of 36.87%.

To the reaction flask, compound 15C (547 mg,1.0 eq), p-toluenesulfonyl hydrazine (337 mg,1.2 eq), 3M HCl in MeOH (0.24 mL,0.5 eq) and methanol (11 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere at 50℃for 6 hours, cooled to room temperature, the reaction solution was concentrated, and purified by silica gel column chromatography to give 480mg of compound 15d in 59.92% yield.

To a reaction flask, compound 15d (480 mg,1.0 eq), compound 1C (298 mg,1.5 eq), potassium carbonate (199mg, 1.6 eq), 2-methyltetrahydrofuran (7.2 mL) and 1, 4-dioxane (2.4 mL) were added in this order, the reaction was allowed to stand for 12 hours under nitrogen atmosphere at 80℃and cooled to room temperature, the reaction solution was extracted with 5% aqueous sodium hydroxide solution, the organic phase was backwashed with water twice, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography with an elution system of dichloromethane/methanol to give 255mg of compound 15 in yield ：53.91％.MS m/z(ESI):523.3012(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝7.97(s,1H),7.14-7.02(m,4H),6.92-6.91(m,1H),6.85–6.83(m,3H),6.71(d,J＝8.0Hz,1H),6.57(t,J＝2.4Hz,1H),6.48-6.41(m,2H),3.96(q,J＝7.2Hz,1H),3.64(d,J＝12.4Hz,2H),3.06(t,J＝4.8Hz,4H),2.69-2.62(m,2H),2.43(t,J＝4.8Hz,5H),2.21(s,3H),1.87–1.82(m,2H),1.57–1.47(m,5H).

Comparative example 1 Compound Ferrostain-1

Compound Ferrostain-1 was purchased from SELLECKCHEM, cat# S7243, lot # S724303.

Preparation of comparative example 2 Compound ref-01

Phenoxazine (2.0 g,1.0 eq) and toluene (20 mL) were added sequentially to the reaction flask, acetyl chloride (1.714g, 2.0 eq) was added thereto, the reaction was continued at 50℃for 1 hour, the temperature was lowered to room temperature, and the reaction solution was concentrated under reduced pressure to obtain compound ref-01a, which was directly used in the next step. MS M/z (ESI): 226.0870 (M+H) ⁺.

To the reaction flask, compound ref-01a (2.618 g,1.0 eq) and dichloromethane (50 mL) were added in this order, the temperature was lowered to 0℃and acetyl chloride (0.895 g,1.0 eq) and aluminum chloride (5.776 g,3.8 eq) were then added, the reaction was stirred for 15min at 0℃and then warmed to room temperature, the reaction was continued for 0.5h at room temperature, the temperature was warmed to 40℃and then allowed to react for 3h at room temperature, the reaction was cooled to room temperature, water-quenching reaction was slowly added, dichloromethane extraction was added, the organic phase was backwashed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated, and silica gel column chromatography was performed to purify 2.01g of compound ref-01b, the yield was 65.97%. MS M/z (ESI): 290.0795 (M+Na) ⁺.

To the reaction flask, compound ref-01b (2.01 g,1.0 eq), absolute ethyl alcohol (40 mL) and concentrated hydrochloric acid (7.5 mL) were added in this order, the reaction was carried out under nitrogen atmosphere at 70℃for 1h, cooled to room temperature, quenched with saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate, the organic phase was backwashed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated to obtain 1.419g of compound ref-01C, yield: 83.77%. MS M/z (ESI): 226.0849 (M+H) ⁺.

To the reaction flask, compound ref-01C (1.319 g,1.0 eq), p-toluenesulfonyl hydrazide (1.408 g,1.2 eq), 3M HCl in MeOH (1.05 mL,0.5 eq) and methanol (28 mL) were added in this order, and the reaction was allowed to stand at 45℃for 3 hours under nitrogen atmosphere, cooled to room temperature, and the reaction solution was concentrated to obtain compound ref-01d, which was directly used in the next step. MS M/z (ESI): 394.1204 (M+H) ⁺.

Sequentially adding a compound ref-01d (500 mg,1.0 eq), a compound 1C (420 mg,1.5 eq), potassium carbonate (270 mg,1.5 eq) and 1, 4-dioxane (25 mL) into a reaction bottle, carrying out heat preservation reaction for 3h at 110 ℃ under a nitrogen environment, cooling to room temperature, quenching reaction liquid by using saturated sodium bicarbonate aqueous solution, adding dichloromethane for extraction, backwashing an organic phase by water twice, drying by anhydrous sodium sulfate, filtering and concentrating, purifying by silica gel column chromatography, eluting by using dichloromethane/methanol to obtain 365mg of the compound ref-01, and obtaining the yield ：74.55％.MS m/z(ESI):386.2234(M+H)⁺.¹HNMR(400MHz,DMSO-d6)δ＝8.10(s,1H),7.05(d,J＝8.4Hz,2H),6.85(d,J＝8.4Hz,2H),6.70(td,J＝7.4,2.0Hz,1H),6.59–6.50(m,3H),6.45(dd,J＝8.0,2.0Hz,1H),6.41(dd,J＝7.6,1.6Hz,1H),6.24(d,J＝2.0Hz,1H),3.83(q,J＝7.2Hz,1H),3.08–3.05(m,4H),2.44–2.42(m,4H),2.21(s,3H),1.44(d,J＝7.2Hz,3H).

Comparative example 3 preparation of Compound ref-02

To a reaction flask, compound 1e (1 g,1.0 eq), 4-bromobenzaldehyde (1.07 g,1.1 eq), tris (dibenzylideneacetone) dipalladium (241 mg,0.05 eq), 4, 5-bis-diphenylphosphine-9, 9-dimethylxanthene (304 mg,0.1 eq), cesium carbonate (2.618 g,1.5 eq) and 1, 4-dioxane (20 mL) were added in this order, and the reaction was carried out under nitrogen atmosphere with a heat-preservation condition at 105℃for 6 hours, TLC showed complete conversion of the starting material, and cooled to room temperature, the reaction solution was filtered through celite, the filtrate was concentrated, and purified by silica gel column chromatography to give 460mg of compound ref-02a in 29.75% yield.

To the reaction flask, compound ref-02a (460 mg,1.0 eq), ethanol (11.5 mL), water (2.3 mL) and tetrahydrofuran (4.6 mL) were sequentially added, hydroxylamine hydrochloride (120 mg,1.1 eq) was further added, the reaction was carried out at room temperature for 20 hours, tlc showed complete conversion of the starting material, and the reaction solution was concentrated to obtain compound ref-02b, which was directly used in the next step.

To the reaction flask, compound ref-02b (284 mg,1.0 eq), ethanol (15 mL) and borane-pyridine (290 mg,2.0 eq) were added sequentially, cooled to 0 ℃, then 10% hydrochloric acid (3.1 mL) was added, the reaction was stirred for 10min at 0 ℃, warmed to room temperature, continued to react for 1h at room temperature, tlc showed complete conversion of the raw materials, the reaction solution was quenched with saturated aqueous sodium bicarbonate, extracted with dichloromethane, the organic phase was backwashed once with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound ref-02C, which was directly used in the next step.

Sequentially adding a compound ref-02C (240 mg,1.0 eq), 2- (4-methylpiperazin-1-yl) acetic acid (160 mg,1.3 eq) and N, N-dimethylformamide (2.5 mL) into a reaction bottle, cooling to 0 ℃, adding N, N-diisopropylethylamine (130 mg,1.3 eq), stirring at 0 ℃ for reaction for 10min, adding DMT-MM (280 mg,1.3 eq), continuously stirring at 0 ℃ for reaction for 30min, heating to room temperature, continuously reacting at room temperature for 1.5h, TLC (thin layer chromatography) shows complete conversion of raw materials, adding water for quenching reaction, adding dichloromethane for extraction, backwashing an organic phase by saturated sodium chloride aqueous solution, drying by anhydrous sodium sulfate, filtering and concentrating, purifying by silica gel column chromatography, eluting the system by dichloromethane/methanol to obtain 108mg of the compound ref-02, and obtaining the yield ：31.01％.MS m/z(ESI):452.3025(M+H)⁺.¹HNMR(400MHz,MeOH-d4)δ＝7.16–6.92(m,8H),4.65(s,2H),3.16(d,J＝11.2Hz,3H),2.67–2.59(m,9H),2.33(s,3H),1.78(d,J＝12.4Hz,2H),1.50–1.24(m,5H),1.01(s,3H).

Study of the inhibition ratio of the Compounds of Experimental examples to Ferroptosis

In order to study the inhibition rate of the compound to Ferroptosis, a Ferroptosis screening model is constructed, and the specific method is that a Ferroptosis screening model mainly adopts a CCK8 cell viability detection method. Human fibrosarcoma cells HT1080 (China academy of sciences cell bank, TCHu) and human neuroblastoma cells SH-SY5Y (China academy of sciences cell bank, TCHu 97) were first cultured in dishes, wherein HT1080 cell culture medium was MEM+10% FBS+2mM L-Glutamine, and SH-SY5Y cell culture medium was DMEM+10% FBS. Cells in the logarithmic growth phase were seeded in 96-well plates (5000 cells/well) at 80. Mu.L per well and then placed in incubator at 37℃in 5% CO ₂ for culture to allow the cells to adhere. After 24h, 10. Mu.L of the compound of the invention (10. Mu.M or 1. Mu.M starting, 3-fold dilution, 8 concentrations) and 10. Mu.L of Ferroptosis inducer Erastin (Selleck, S7242) at a final concentration of 10. Mu.M were added at various concentrations prepared using cell culture medium, 3 multiplex wells were set per compound to ensure accuracy of the results. Meanwhile, a positive control group (initial 1 mu M concentration prepared by adding 10 mu L of the same culture medium, 3 times dilution, 8 concentrations of compounds Ferrostain-1, ref-01 and ref-02), a blank control group (adding the specified equal volume cell culture medium containing the same DMSO concentration and containing no cells) and a solvent control group (adding the specified equal volume cell culture medium containing the same DMSO concentration and containing cells) are arranged, and 3 duplicate wells are arranged in each group, so that the accuracy of the result is ensured. After dosing, the mixture is put into a incubator and cultured for 24 hours. mu.L of CCK8 solution (Topscience, C0005) was added to each well, incubated in incubator for 2-4 hours, and absorbance at 450nm was measured with a microplate reader to calculate the inhibition of Ferroptosis by the compound. The inhibition ratio was calculated using the following formula (inhibitionrate, IR) = [1- (a experimental group-a blank)/(a solvent-a blank) ]x100%

Inhibition rate change curves were fitted using GraphPadPrism6 software and IC ₅₀ values calculated.

Experimental results show that the compounds 1-15 have good inhibitory activity on iron death of HT1080 cells and SH-SY5Y cells, the IC ₅₀ values are lower than 200nM, and partial activity results of the structures such as the compound 1 are shown in Table 1.

Table 1 results of iron death inhibitory Activity of partial Compounds on HT1080 cells and SH-SY5Y cells

As shown in Table 1, the compounds of the present invention have excellent iron-death suppressing activity, and the structures of compound 1 and the like are significantly superior to those of conventional control compounds Ferrostain-1, ref-01 and ref-02, for example, 11.6 to 31.4 times as high as Ferrostain-1, 9.4 to 25.4 times as high as ref-01 and 12.9 to 35.1 times as high as ref-02 in terms of the inhibitory activity of HT1080 cells, and 6.9 to 12.2 times as high as Ferrostain-1, 7.9 to 13.9 times as high as ref-01 and 8.0 to 14.0 times as high as ref-02 in terms of the inhibitory activity of SH-SY5Y cells in terms of compound 1, compound 3 and compound 4. Thus, the compounds of the present invention can be used for preparing iron death inhibitors to prevent and/or treat various diseases related to iron death, such as neurodegenerative diseases, parkinsonism, cerebral apoplexy, cardiovascular diseases, renal failure, diabetic complications, cancers and the like.

Unless otherwise defined, all terms used herein are intended to have the meanings commonly understood by those skilled in the art.

The described embodiments of the present invention are intended to be illustrative only and not to limit the scope of the invention, and various other alternatives, modifications, and improvements may be made by those skilled in the art within the scope of the invention, and therefore the invention is not limited to the above embodiments but only by the claims.

Claims (10)

1. A compound with a structure shown as a formula (I) or pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof,

In formula (I), ring a is selected from C ₆～C₁₂ aryl optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

Ring B is selected from a 5-12 membered heterocyclyl or a 5-12 membered heteroaryl, said heterocyclyl or heteroaryl comprising at least one N atom as a ring atom, and said heterocyclyl or heteroaryl being optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy;

Each R ₁、R₂ is independently selected from hydrogen, C ₁～C₆ alkyl, or C ₃～C₁₀ cycloalkyl;

Each R ₃ is independently selected from hydrogen, halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ haloalkoxy, C ₃～C₁₀ cycloalkyl, 5-to 12-membered heterocyclyl, C ₆～C₁₂ aryl, or 5-to 12-membered heteroaryl, said hydroxy, carboxy, amino, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, or heteroaryl being optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, cyano, amino, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

m is selected from 0, 1,2, 3 or 4.

2. The compound of claim 1, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, wherein in formula (I),

Ring a is selected from C ₆～C₁₀ aryl;

Ring B is selected from a 5-8 membered heterocyclyl group comprising 1-3N atoms as ring atoms, and which heterocyclyl group is optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy;

Each R ₁、R₂ is independently selected from hydrogen or C ₁～C₆ alkyl;

Each R ₃ is independently selected from hydrogen, halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, C ₁～C₆ haloalkoxy, 5-to 8-membered heterocyclyl, or 5-to 8-membered heteroaryl, optionally substituted with one or more substituents selected from halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy;

m is selected from 0,1, 2 or 3.

3. The compound of claim 2, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, wherein in formula (I),

Ring a is selected from phenyl;

Ring B is selected from a 5-6 membered heterocyclic group containing 1-2N atoms as ring atoms, and optionally substituted with one or more substituents selected from C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy or C ₁～C₄ haloalkoxy, said heterocyclic group preferably being selected from piperidinyl or piperazinyl;

Each R ₁、R₂ is independently selected from hydrogen or C ₁～C₄ alkyl;

Each R ₃ is independently selected from hydrogen, F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, C ₁～C₄ haloalkoxy, or a 5-to 6-membered heterocyclyl containing at least one N atom as a ring atom, and the heterocyclyl is optionally substituted with one or more substituents selected from F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, or C ₁～C₄ haloalkoxy;

m is selected from 0, 1 or 2.

4. A compound according to any one of claims 1-3, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, or isotopic label thereof, wherein the compound has a structure as shown in formula (II):

In formula (II), each Z ₁、Z₂、Z₃ is independently selected from-C (R ₄)₂-、-NR₄ -or a bond;

Each R ₄ is independently selected from hydrogen, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy, or C ₁～C₆ haloalkoxy, preferably selected from hydrogen, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy, or C ₁～C₄ haloalkoxy;

R ₁、R₂、R₃, m are each independently as defined in any one of claims 1 to 3;

preferably, the compound has a structure as shown in formula (III):

in formula (III), R ₁、R₂、R₃、R₄, m are each independently defined as in formula (II).

5. The compound of claim 4, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, or isotopic label thereof, wherein the compound has a structure according to formula (IV):

in the formula (IV), Y is selected from-C (R ₆)₂-、-NR₆ -, -O-or-S-;

R ₅、R₆ is each independently selected from hydrogen, halogen, hydroxy, carboxy, C ₁～C₆ alkyl, C ₁～C₆ haloalkyl, C ₁～C₆ alkoxy or C ₁～C₆ haloalkoxy, preferably selected from hydrogen, F, cl, hydroxy, carboxy, C ₁～C₄ alkyl, C ₁～C₄ haloalkyl, C ₁～C₄ alkoxy or C ₁～C₄ haloalkoxy;

n is selected from 0, 1 or 2;

Each R ₁、R₂、R₄ is independently as defined in claim 4;

preferably, the compound has a structure as shown in formula (IV-1) or formula (IV-2):

In formula (IV-1) or formula (IV-2), Y, R ₁、R₂、R₄、R₅, n are each independently as defined in formula (IV);

more preferably, each R ₁、R₂、R₄ is independently selected from hydrogen, methyl, ethyl, n-propyl or isopropyl.

6. A compound or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, wherein the compound comprises:

7. The method for preparing the compound or the pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof according to claim 1, wherein the preparation method comprises the steps of reacting an aniline compound with a structure shown in a formula (I-1) with a halogenide with a structure shown in a formula (I-2) to obtain a first intermediate with a structure shown in a formula (I-3), reacting the first intermediate with p-toluenesulfonyl hydrazine to obtain a second intermediate with a structure shown in a formula (I-4), and reacting the second intermediate with a boric acid compound with a structure shown in a formula (I-5) to obtain the compound with the structure shown in the formula (I);

Wherein X is selected from halogen, preferably from Br;

ring a, ring B, R ₁、R₂、R₃, m are each independently as defined in claim 1.

8. A pharmaceutical composition comprising a compound of any one of claims 1-6, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate, or isotopic label thereof, and one or more pharmaceutically acceptable excipients.

9. Use of a compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, or a pharmaceutical composition according to claim 8, for the preparation of an iron death inhibitor.

10. Use of a compound according to any one of claims 1-6, or a pharmaceutically acceptable salt, stereoisomer, solvate, hydrate or isotopic label thereof, or a pharmaceutical composition according to claim 8, for the manufacture of a medicament for the prevention and/or treatment of neurodegenerative diseases, tissue ischemia reperfusion injury, stroke, cardiovascular disease, renal failure, liver injury, diabetic complications or cancer;

Preferably, the neurodegenerative disease is parkinsonism or alzheimer's disease;

Preferably, the cerebral apoplexy is ischemic cerebral apoplexy or hemorrhagic cerebral apoplexy;

Preferably, the cancer is pancreatic cancer.