HK40009413B

HK40009413B - Hdac6 selective inhibitors, preparation method therefor, and application thereof

Info

Publication number: HK40009413B
Application number: HK19132738.6A
Authority: HK
Inventors: 吴颢; 韦昌青; 郭强; 张贵芬; 刘斌; 廖勇刚; 肖瑶; 陈曙辉
Original assignee: 基石药业（苏州）有限公司; 拓石药业（上海）有限公司; 基石药业
Priority date: 2017-01-10
Filing date: 2018-01-10
Publication date: 2023-03-10

Description

HDAC6 selective inhibitor and preparation method and application thereof

Cross Reference to Related Applications

The present application claims priority of the chinese patent application CN201710017287.2 filed 2017.01.10, the contents of which are incorporated herein.

Technical Field

The invention relates to a compound serving as a selective inhibitor of histone deacetylase 6 (HDAC 6) and application thereof in preparing a medicament for treating HDAC6 related diseases. In particular to a compound shown as a formula (I) and pharmaceutically acceptable salts thereof.

Background

The WHO experts predict that the world population will reach 80 hundred million in 2020, the cancer incidence will reach 2000 million, and the death will reach 1200 million. Cancer will become the first killer of humans in the new century, and constitutes the most serious threat to human survival. China, which is in the process of industrialization, is the second cancer-high-incidence country in the world after the United states, and the cancer incidence and mortality rate of China show a remarkable rising trend. Urban cancers account for the first and rural cancers for the second of the overall cause of death. With the rapid increase of cancer morbidity and mortality in China, the medical cost for cancer is over 1,500 billion yuan per year in China.

HDAC inhibitors are widely used in a variety of cancers and can be combined with a variety of drugs to enhance the therapeutic effect of the drugs, and are well-established anti-tumor targets. Histone Deacetylases (HDACs) and Histone Acetyltransferases (HATs) co-regulate transcription of genes within the nucleus. In cancer cells, overexpression of HDACs leads to increased deacetylation, which increases the attraction between DNA and histones, making nucleosomes very compact and detrimental to the expression of tumor suppressor genes. The inhibitor (HDACI) can regulate the expression of apoptosis and differentiation related proteins by improving histone acetylation, induce apoptosis and differentiation, and become a new anti-tumor drug. Moreover, HDAC is also involved in the regulation of many metabolic diseases, such as Alzheimer's disease, parkinson's disease and other diseases, and HDACi inhibitors show good effects in both animal and human experiments.

Of the 18 deacetylase subtypes in total, HDAC6 is the only deacetylase subtype found in the cytoplasm, while the other 17 HDACs are present in the nucleus. HDAC6 does not directly catalyze histones, but rather takes tubulin (tubulin) and heat shock protein (Hsp 90) as substrates through which cell trafficking, adhesion and motility are regulated (i.e., no gene regulation). Current clinical experimental results have demonstrated that HDAC6 selective inhibitors are safe and effective (POC). Clinical studies with the first HDAC6 selective inhibitor, ACY-1215 (acetolon), demonstrated that the selective HDAC6 inhibitor has greater safety and thus greater commercial potential.

Proteasome inhibitors (Drug Design, development and Therapy 2016, 10-217-226) can block the degradation of a number of regulatory proteins, causing the disruption and overloading of intracellular signaling systems, resulting in the inhibition of cell growth, and ultimately in the slowing or even halting of tumor progression. The HDAC inhibitor is widely applied to various cancers and can be combined with various medicaments to enhance the treatment effect of the medicaments, for example, the combination of the HDAC inhibitor panobinostat and a proteasome inhibitor bortezomib can increase the curative effect of multiple myeloma and obviously reduce the toxicity.

Disclosure of Invention

The invention provides a compound shown as a formula (I), pharmaceutically acceptable salts and isomers thereof:

selected from: a single bond or a double bond;

n is selected from: 0 or 1;

T ₁ 、T ₂ each independently selected from: CH. CH (CH) ₂ 、-C(＝O)-、N；

T ₃ Selected from: c or N;

Z ₁ 、Z ₂ 、Z ₃ each independently selected from: CH or N;

L ₁ selected from: a single bond, -NH-, -C (= O) -NH-;

R ₁ selected from optionally substituted with 1,2 or 3R: c _1-3 Alkyl, phenyl, 6-membered heteroaryl;

R ₂ selected from: H. f, cl, br, I;

the A ring is selected from: 4-7 membered heterocycloalkyl;

r is selected from: F. cl, br, I;

said "hetero" of 6-membered heteroaryl, 4-7-membered heterocycloalkyl are each independently selected from: -NH-, N, -O-;

in any of the above cases, the number of heteroatoms or heteroatom groups is independently selected from 1,2 or 3, respectively.

In some embodiments of the invention, R is as defined above ₁ Selected from the group consisting of optionally substituted with 1,2 or 3R: methyl, ethyl, isopropyl, phenyl, pyridyl.

In some embodiments of the invention, R is as defined above ₁ Selected from the group consisting of optionally substituted with 1,2 or 3R: CH (CH) ₃ 、

In some embodiments of the present invention, R is ₁ Selected from: CH (CH) ₃ 、

In some embodiments of the invention, the above A ring is selected from: oxetanyl, tetrahydrofuryl, tetrahydropyranyl, 1, 3-dioxolanyl, 1, 4-dioxacycloheptyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, morpholinyl.

In some embodiments of the invention, the above A ring is selected from:

in some embodiments of the present invention, the structural unitSelected from:

in some embodiments of the invention, the structural unitSelected from:

in some embodiments of the invention, the structural unitSelected from the group consisting of:

in some embodiments of the present invention, the structural unitSelected from the group consisting of:

in some embodiments of the present invention, the structural unitSelected from:

in some embodiments of the invention, the structural unitSelected from the group consisting of: -CH ₂ -、-NH-、-C(＝O)-NH-、

In some embodiments of the invention, the structural unitSelected from:

in some embodiments of the invention, the structural unitSelected from:

in some embodiments of the invention, R is as defined above ₁ Selected from optionally substituted with 1,2 or 3R: methyl, ethyl, phenyl, pyridyl, the other variables being as defined herein.

In some embodiments of the invention, R is as defined above ₁ Selected from optionally substituted with 1,2 or 3R: CH (CH) ₃ 、Other variables are as defined herein.

In some embodiments of the invention, R is as defined above ₁ Selected from: CH (CH) ₃ 、Other variables are as defined herein.

In some embodiments of the invention, the above-mentioned ring A is selected from: oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 1, 3-dioxolanyl, 1, 4-dioxacycloheptanyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, morpholinyl, and other variables are as defined herein.

In some embodiments of the invention, the above A ring is selected from:other variables are as defined herein.

In some embodiments of the invention, the structural unitSelected from:other variables are as defined herein.

In some embodiments of the present invention, the structural unitSelected from:the other variables are as defined herein.

In some embodiments of the invention, the structural unitSelected from the group consisting of:other variables are as defined herein.

In some embodiments of the present invention, the structural unitSelected from:other variables are as defined herein.

In some embodiments of the present invention, the structural unitSelected from the group consisting of:the other variables are as defined herein.

In some embodiments of the invention, the structural unitSelected from:the other variables are as defined herein.

In some embodiments of the present invention, the structural unitSelected from: -CH ₂ -、-NH-、-C(＝O)-NH-、Other variables are as defined herein.

Still other embodiments of the present invention are derived from any combination of the above variables.

In some embodiments of the invention, the compounds, pharmaceutically acceptable salts, and isomers thereof, as described above, are selected from the group consisting of:

wherein, A ring, R ₁ 、R ₂ 、L ₁ And n is as defined herein.

In some embodiments of the invention, the compounds, pharmaceutically acceptable salts, and isomers thereof described above are selected from the group consisting of:

wherein the content of the first and second substances,

E ₁ 、E ₂ are each independently selected from-O-, -CH ₂ -and-CH ₂ -CH2-；

R、R ₂ 、L ₁ And n is as defined herein.

The present invention also provides the following compounds and pharmaceutically acceptable salts thereof, selected from:

the invention also provides a pharmaceutical composition, which comprises the compound or the pharmaceutically acceptable salt thereof with effective treatment amount as an active ingredient and a pharmaceutically acceptable carrier.

The invention also provides application of the compound or the pharmaceutically acceptable salt thereof in preparing a medicament for treating HDAC6 related diseases.

The invention also provides application of the composition in preparing a medicament for treating HDAC6 related diseases.

In some embodiments of the invention, the above-mentioned use, wherein the medicament is a medicament for treating multiple myeloma.

Technical effects

As a novel histone deacetylase 6 (HDAC 6) selective inhibitor, the compound has remarkable in vitro activity, has remarkable inhibition effect on HDAC6 enzyme, and has the characteristic of high selectivity on weak inhibition on HDAC 1; in addition, administration in combination with Ixazomib increases the efficacy of multiple myeloma and significantly reduces toxicity.

Definitions and explanations

As used herein, the following terms and phrases are intended to have the following meanings, unless otherwise indicated. A particular term or phrase, unless specifically defined, should not be considered as indefinite or unclear, but rather construed according to ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient. The term "pharmaceutically acceptable" as used herein is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from the compounds of the present invention found to have particular substituents, with relatively nontoxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts with inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like; and salts of organic acids including acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, methanesulfonic, and the like; also included are Salts of amino acids such as arginine, etc., and Salts of organic acids such as glucuronic acid (see Berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science 66 (1977). Certain specific compounds of the invention contain both basic and acidic functionalities and can thus be converted to either base or acid addition salts.

Preferably, the neutral form of the compound is regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms by certain physical properties, such as solubility in polar solvents.

As used herein, "pharmaceutically acceptable salts" are derivatives of the compounds of the present invention wherein the parent compound is modified by salification with an acid or salification with a base. Examples of pharmaceutically acceptable salts include, but are not limited to: inorganic or organic acid salts of bases such as amines, alkali metal or organic salts of acid groups such as carboxylic acids, and the like. Pharmaceutically acceptable salts include the conventional non-toxic salts or the quaternary ammonium salts of the parent compound, for example, salts formed with non-toxic inorganic or organic acids. Conventional non-toxic salts include, but are not limited to, those derived from inorganic or organic acids selected from the group consisting of 2-acetoxybenzoic acid, 2-hydroxyethanesulfonic acid, acetic acid, ascorbic acid, benzenesulfonic acid, benzoic acid, bicarbonate, carbonic acid, citric acid, edetic acid, ethanedisulfonic acid, ethanesulfonic acid, fumaric acid, glucoheptose, gluconic acid, glutamic acid, glycolic acid, hydrobromic acid, hydrochloric acid, hydroiodide, hydroxyl, hydroxynaphthalene, isethionic acid, lactic acid, lactose, dodecylsulfonic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, nitric acid, oxalic acid, pamoic acid, pantothenic acid, phenylacetic acid, phosphoric acid, polygalacturonic acid, propionic acid, salicylic acid, stearic acid, glycolic acid, succinic acid, sulfamic acid, sulfanilic acid, sulfuric acid, tannin, tartaric acid, and p-toluenesulfonic acid.

The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

In addition to salt forms, the compounds provided herein also exist in prodrug forms. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to convert to the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention in an in vivo environment by chemical or biochemical means.

Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, the solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention.

Certain compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereomers, geometric isomers and individual isomers are all included within the scope of the present invention.

Using wedge and dashed bonds, unless otherwise indicatedRepresenting an absolute configuration of a solid centre, by wavy linesRepresenting wedge or dashed bondsBy usingShowing the relative configuration of the stereocenters. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, they include E, Z geometric isomers unless otherwise specified. Likewise, all tautomeric forms are included within the scope of the invention.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-) -and (+) -enantiomers, (R) -and (S) -enantiomers, diastereomers, (D) -isomers, (L) -isomers, as well as racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Optically active (R) -and (S) -isomers as well as D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the invention is desired, it may be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to afford the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and the pure enantiomers are recovered. Furthermore, separation of enantiomers and diastereomers is typically accomplished by using chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., carbamate formation from amines).

Compounds of the inventionNon-natural proportions of atomic isotopes may be contained at one or more of the atoms constituting such compounds. For example, the compound may be labeled with a radioisotope, such as tritium (a) ³ H) Iodine-125 ( ¹²⁵ I) Or C-14 ( ¹⁴ C) In that respect All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention. "optional" or "optionally" means that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent, and may include variations of deuterium and hydrogen, so long as the valency of the particular atom is normal and the substituted compound is stable. When the substituent is keto (i.e = O), it means that two hydrogen atoms are substituted. The keto substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted and, unless otherwise specified, the type and number of substituents may be arbitrary on the basis of chemical feasibility.

When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of one linking group is 0, e.g. - (CRR) ₀ -, represents that the linking group is a single bond.

When one of the variables is selected from a single bond, it means that the two groups to which it is attached are directly connected, for example, where L represents a single bond in A-L-Z means that the structure is actually A-Z.

When a substituent is absent, it indicates that the substituent is absent, e.g., when X is absent in A-X, it indicates that the structure is actually A. When one isWhen the bonds of a substituent may be cross-linked to two atoms on a ring, such substituent may be bonded to any atom on the ring. When the substituents listed are not indicated by which atom they are attached to the compounds included in the general chemical structure formula but not specifically mentioned, such substituents may be bonded through any atom thereof. Combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds. For example, a structural unitMeaning that it may be substituted at any position on the cyclohexyl or cyclohexadiene.

Unless otherwise specified, the term "hetero" means a heteroatom or a group of heteroatoms (i.e., a group of atoms containing a heteroatom), including atoms other than carbon (C) and hydrogen (H) and groups of atoms containing such heteroatoms, for example, oxygen (O), nitrogen (N), sulfur (S), silicon (Si), germanium (Ge), aluminum (Al), boron (B), -O-, -S-, = O, = S, -C (= O) O-, -C (= O) -, -C (= S) -, -S (= O) 2-, and optionally substituted-C (= O) N (H) -, -C (= NH) -, -S (= O) ₂ N (H) -or-S (= O) N (H) -.

Unless otherwise specified, "ring" means a substituted or unsubstituted cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, aryl, or heteroaryl group. The term "ring" includes monocyclic, bicyclic, spiro, fused or bridged rings. The number of atoms in the ring is generally defined as the number of ring members, e.g., "5 to 7 membered ring" means 5 to 7 atoms arranged around the ring. Unless otherwise specified, the ring optionally contains 1 to 3 heteroatoms. Thus, "5-to 7-membered ring" includes, for example, phenyl, pyridine and piperidinyl; on the other hand, the term "5-to 7-membered heterocycloalkyl ring" includes pyridyl and piperidyl, but does not include phenyl. The term "ring" also includes ring systems containing at least one ring, each of which independently conforms to the above definition.

Unless otherwise specified, the term "heterocycle" or "heterocyclyl" means a stable heteroatom or heteroatom group containing monocyclic, bicyclic, or tricyclic ring which may be saturated, partially unsaturated, or unsaturated (aromatic), which comprises carbon atoms and 1,2,3, or 4 ring heteroatoms independently selected from N, O, and S, wherein any of the above heterocycles may be fused to a benzene ring to form a bicyclic ring. The nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S (O) p, p being 1 or 2). The nitrogen atom may be substituted or unsubstituted (i.e., N or NR, where R is H or other substituents already defined herein). The heterocyclic ring may be attached to any heteroatom or carbon pendant group to form a stable structure. The heterocyclic rings described herein may be substituted at the carbon or nitrogen position if the resulting compound is stable. The nitrogen atoms in the heterocycle are optionally quaternized. In a preferred embodiment, when the total number of S and O atoms in the heterocycle exceeds 1, the heteroatoms are not adjacent to one another. Another preferred embodiment is that the total number of S and O atoms in the heterocycle does not exceed 1. As used herein, the term "aromatic heterocyclic group" or "heteroaryl" means a stable 5,6, 7 membered monocyclic or bicyclic or 7, 8, 9 or 10 membered bicyclic heterocyclic group aromatic ring comprising carbon atoms and 1,2,3 or 4 ring heteroatoms independently selected from N, O and S. The nitrogen atom may be substituted or unsubstituted (i.e. N or NR, wherein R is H or other substituents already defined herein). The nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S (O) p, p being 1 or 2). It is noted that the total number of S and O atoms on the heteroaromatic ring does not exceed 1. Bridged rings are also included in the definition of heterocyclic. Bridged rings are formed when one or more atoms (i.e., C, O, N, or S) connect two non-adjacent carbon or nitrogen atoms. Preferred bridge rings include, but are not limited to: one carbon atom, two carbon atoms, one nitrogen atom, two nitrogen atoms, and one carbon-nitrogen group. It is worth noting that a bridge always converts a single ring into a three ring. In bridged rings, ring substituents may also be present on the bridge.

Examples of heterocyclic compounds include, but are not limited to: acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzomercaptofuranyl, benzomercaptophenyl, benzoxazolyl, benzoxazolinyl, benzothiazolyl, benzotriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4 aH-carbazolyl, carbolinyl, chromanyl, chromene, cinnolinyl decahydroquinolinyl, 2H,6H-1,5, 2-dithiazinyl, dihydrofuro [2,3-b ] tetrahydrofuranyl, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolylene, indolinyl, indolizinyl, indolyl, 3H-indolyl, isobenzofuranyl, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,3, 4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazine, phenothiazine, benzoxanthine, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, piperidyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2, 5-thiadiazinyl, 1,2, 3-thiadiazolyl, 1,2, 4-thiadiazolyl, 1,2, 5-thiadiazolyl, 1,3, 4-thiadiazolyl, thianthrenyl, thiazolyl, isothiazolylthio, thienooxazolyl, thienothiazolyl, thienoimidazolyl, thienyl, triazinyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, 1,2, 5-triazolyl, 1,3, 4-triazolyl, and xanthenyl. Fused ring and spiro compounds are also included.

Unless otherwise specified, the term "hydrocarbyl" or its derivatives (e.g., alkyl, alkenyl, alkynyl, aryl, etc.) by itself or as part of another substituent, means a straight, branched, or cyclic hydrocarbon radical, or combinations thereof, that may be fully saturated (e.g., alkyl), mono-or poly-unsaturated (e.g., alkenyl, alkynyl, aryl), that may be mono-or poly-substituted, that may be mono (e.g., methyl), di (e.g., methylene), or multi (e.g., monovalent (e.g., methyl), di (e.g., methylene), or poly (e.g., methylene)Methine group) which may include divalent or polyvalent radicals having a specified number of carbon atoms (e.g., C) ₁ -C ₁₂ Represents 1 to 12 carbons, C _1-12 Is selected from C ₁ 、C ₂ 、C ₃ 、C ₄ 、C ₅ 、C ₆ 、C ₇ 、C ₈ 、C ₉ 、C ₁₀ 、C ₁₁ And C ₁₂ ；C _3-12 Is selected from C ₃ 、C ₄ 、C ₅ 、C ₆ 、C ₇ 、C ₈ 、C ₉ 、C ₁₀ 、C ₁₁ And C ₁₂ . ). "hydrocarbyl" includes, but is not limited to, aliphatic hydrocarbyl including linear and cyclic, specifically including, but not limited to alkyl, alkenyl, alkynyl, and aromatic hydrocarbyl including, but not limited to, 6-12 membered aromatic hydrocarbyl such as benzene, naphthalene, and the like. In some embodiments, the term "hydrocarbyl" denotes a straight or branched chain radical or a combination thereof, which may be fully saturated, mono-or polyunsaturated, and may include divalent and polyvalent radicals. Examples of saturated hydrocarbon radicals include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, isobutyl, cyclohexyl, (cyclohexyl) methyl, cyclopropylmethyl, and homologs or isomers of radicals such as n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. Unsaturated hydrocarbon groups have one or more double or triple bonds, examples of which include, but are not limited to, ethenyl, 2-propenyl, butenyl, crotyl, 2-isopentenyl, 2- (butadienyl), 2, 4-pentadienyl, 3- (1, 4-pentadienyl), ethynyl, 1-and 3-propynyl, 3-butynyl, and higher homologs and isomers.

Unless otherwise specified, the term "heterohydrocarbyl" or a subset thereof (such as heteroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl, and the like) by itself or in combination with another term means a stable straight-chain, branched, or cyclic hydrocarbon radical, or combination thereof, consisting of a number of carbon atoms and at least one heteroatom. In some embodiments, the term "heteroalkyl," by itself or in combination with another term, means a stable straight-chain, branched-chain hydrocarbon radical, or combination thereof, having a number of carbonsAtoms and at least one heteroatom. In a typical embodiment, the heteroatoms are selected from B, O, N and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen heteroatom is optionally quaternized. The heteroatom or heteroatom group may be located at any internal position of the heterohydrocarbyl group, including the position at which the hydrocarbyl group is attached to the remainder of the molecule, but the terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional expressions to refer to those alkyl groups that are attached to the remainder of the molecule through an oxygen atom, an amino group, or a sulfur atom, respectively. Examples include, but are not limited to-CH ₂ -CH ₂ -O-CH ₃ 、-CH ₂ -CH ₂ -NH-CH ₃ 、-CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ 、-CH ₂ -S-CH ₂ -CH ₃ 、-CH ₂ -CH ₂ 、-S(O)-CH ₃ 、-CH ₂ -CH ₂ -S(O) ₂ -CH ₃ 、-CH＝CH-O-CH ₃ 、-CH ₂ -CH＝N-OCH ₃ and-CH = CH-N (CH) ₃ )-CH ₃ . Up to two heteroatoms may be consecutive, e.g. -CH ₂ -NH-OCH ₃ 。

Unless otherwise specified, the terms "cycloalkyl," "heterocycloalkyl," or subset thereof (e.g., aryl, heteroaryl, cycloalkyl, heterocycloalkyl, cycloalkenyl, heterocycloalkenyl, cycloalkynyl, heterocycloalkynyl, etc.) by themselves or in combination with other terms, mean a cyclized "hydrocarbyl," "heterocarbyl," respectively. Furthermore, in the case of heterohydrocarbyl or heterocycloalkcarbyl (e.g., heteroalkyl, heterocycloalkyl), the heteroatom may occupy a position where the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl groups include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Non-limiting examples of heterocyclyl groups include 1- (1, 2,5, 6-tetrahydropyridinyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran indol-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, and 2-piperazinyl.

Unless otherwise specified, the term "alkyl" is intended to mean a straight-chain or branched-chain saturated hydrocarbon radical, which may be monosubstituted (e.g., -CH) ₂ F) Or polysubstituted (e.g. -CF) ₃ ) And may be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). Examples of alkyl groups include methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, s-butyl, t-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), and the like.

Unless otherwise specified, "alkenyl" refers to an alkyl group having one or more carbon-carbon double bonds at any position in the chain, which may be mono-or poly-substituted, and which may be mono-, di-or polyvalent. Examples of alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, piperylene, hexadienyl, and the like.

Unless otherwise specified, "alkynyl" refers to an alkyl group having one or more carbon-carbon triple bonds at any position in the chain, and may be mono-or poly-substituted, and may be monovalent, divalent, or polyvalent. Examples of alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, and the like.

Unless otherwise specified, cycloalkyl includes any stable cyclic or polycyclic hydrocarbon group, any carbon atom of which is saturated, which may be mono-or poly-substituted, which may be mono-, di-or polyvalent. Examples of such cycloalkyl groups include, but are not limited to, cyclopropyl, norbornyl, [2.2.2] bicyclooctane, [4.4.0] bicyclodecane, and the like.

Unless otherwise specified, cycloalkenyl includes any stable cyclic or polycyclic hydrocarbon radical containing one or more unsaturated carbon-carbon double bonds at any position on the ring, which may be mono-or polysubstituted, and which may be mono-, di-or polyvalent. Examples of such cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, and the like.

Unless otherwise specified, cycloalkynyl includes any stable cyclic or polycyclic hydrocarbon radical containing one or more carbon-carbon triple bonds at any position in the ring, which may be mono-or poly-substituted, and which may be mono-, di-or polyvalent.

Unless otherwise specified, the term "halo" or "halogen" by itself or as part of another substituent meansFluorine, chlorine, bromine or iodine atoms. Furthermore, the term "haloalkyl" is intended to include monohaloalkyl and polyhaloalkyl. For example, the term "halo (C) ₁ -C ₄ ) Alkyl "is intended to include, but not be limited to, trifluoromethyl, 2-trifluoroethyl, 4-chlorobutyl, and 3-bromopropyl, and the like. Unless otherwise specified, examples of haloalkyl include, but are not limited to: trifluoromethyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl.

"alkoxy" represents the above alkyl group having the specified number of carbon atoms attached through an oxygen bridge, unless otherwise specified, C _1-6 Alkoxy radicals comprising C ₁ 、C ₂ 、C ₃ 、C ₄ 、C ₅ And C ₆ Alkoxy group of (2). Examples of alkoxy groups include, but are not limited to: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy and S-pentoxy.

Unless otherwise specified, the term "aryl" means a polyunsaturated aromatic hydrocarbon substituent, which may be mono-or polysubstituted, and which may be mono-, di-or polyvalent, and which may be mono-or polycyclic (e.g., 1 to 3 rings; wherein at least one ring is aromatic), fused together or linked covalently. The term "heteroaryl" refers to an aryl (or ring) containing one to four heteroatoms. In one illustrative example, the heteroatoms are selected from B, N, O, and S, wherein the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atoms are optionally quaternized. The heteroaryl group may be attached to the rest of the molecule through a heteroatom. Non-limiting examples of aryl or heteroaryl include phenyl, naphthyl, biphenyl, pyrrolyl, pyrazolyl, imidazolyl, pyrazinyl, oxazolyl, phenyl-oxazolyl, isoxazolyl, thiazolyl, furyl, thienyl, pyridyl, pyrimidinyl, benzothiazolyl, purinyl, benzimidazolyl, indolyl, isoquinolyl, quinoxalinyl, quinolyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidinyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-quinoxalyl, 5-isoquinolyl, 5-quinoxalinyl, and 6-isoquinolyl. The substituents for any of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below.

Unless otherwise specified, aryl when used in combination with other terms (e.g., aryloxy, arylthio, aralkyl) includes aryl and heteroaryl rings as defined above. Thus, the term "aralkyl" is intended to include those radicals in which an aryl group is attached to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl and the like), including those alkyl groups in which a carbon atom (e.g., methylene) has been replaced by, for example, an oxygen atom, such as phenoxymethyl, 2-pyridyloxymethyl 3- (1-naphthyloxy) propyl and the like.

The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., an affinity substitution reaction). For example, representative leaving groups include triflate; chlorine, bromine, iodine; sulfonate groups such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate and the like; acyloxy groups such as acetoxy, trifluoroacetyloxy, and the like.

The term "protecting group" includes, but is not limited to "amino protecting group", "hydroxyl protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for use in preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: a formyl group; acyl, for example alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyls such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl groups such as benzyl (Bn), trityl (Tr), 1-bis- (4' -methoxyphenyl) methyl; silyl groups, such as Trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS), and the like. The term "hydroxy protecting group" refers to a protecting group suitable for use in preventing side reactions of a hydroxy group. Representative hydroxy protecting groups include, but are not limited to: alkyl groups such as methyl, ethyl and tert-butyl; acyl groups, such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (benzhydryl, DPM); silyl groups, such as Trimethylsilyl (TMS) and t-butyldimethylsilyl (TBS), and the like.

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combinations thereof with other chemical synthetic methods, and equivalents thereof known to those skilled in the art, with preferred embodiments including, but not limited to, examples of the present invention.

The solvent used in the present invention can be commercially available. The invention employs the following abbreviations: aq represents water; HATU represents O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium hexafluorophosphate; EDC stands for N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride; m-CPBA represents 3-chloroperoxybenzoic acid; eq represents equivalent, equivalent; CDI represents carbonyldiimidazole; DCM represents dichloromethane; PE represents petroleum ether; DIAD represents diisopropyl azodicarboxylate; DMF represents N, N-dimethylformamide; DMSO represents dimethyl sulfoxide; etOAc for ethyl acetate; etOH represents ethanol; meOH represents methanol; CBz represents benzyloxycarbonyl, an amine protecting group; BOC represents tert-butylcarbonyl which is an amine protecting group; HOAc represents acetic acid; naCNBH ₃ Represents sodium cyanoborohydride; r.t. represents room temperature; O/N stands for overnight; THF represents tetrahydrofuran; boc ₂ O represents di-tert-butyl dicarbonate; TFA represents trifluoroacetic acid; DIPEA stands for diisopropylethylamine; SOCl ₂ Represents thionyl chloride; CS ₂ Represents carbon disulfide; tsOH represents p-toluenesulfonic acid; NFSI represents N-fluoro-N- (phenylsulfonyl) benzenesulfonamide; NCS represents 1-chloropyrrolidine-2, 5-dione; n-Bu ₄ NF represents tetrabutyl ammonium fluoride; iPrOH represents 2-propanol; mp represents melting point; LDA represents lithium diisopropylamide; EA represents ethyl acetate; DPPF represents 1,1' -bis diphenylphosphinoferrocene; et represents ethyl; me represents a methyl group; DCM represents dichloromethane; TMSCHN ₂ Represents trimethylsilylated diazomethane; DCE represents dichloroethane; BSA represents bovine serum albumin; TCEP represents tris (2-carboxyethyl) phosphine; BH ₃ -Me ₂ S represents borane methyl sulfide; pd (OAc) ₂ Represents palladium acetate; DPPP stands for 1, 3-bis (diphenylphosphino) propane; TEA stands for triethylamine; TMSCl represents trimethylchlorosilane; EDCI represents 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride; et (ethyl acetate) ₃ N represents triethylamine; meI represents methyl iodide; KHMDS represents potassium hexamethyldisilazide; n-BuLi represents n-butyllithium; pd ₂ (dba) ₃ Represents tris (dibenzylideneacetone) dipalladium; t-BuXPhOS represents 2-di-tert-butylphosphino-2 ',4',6' -triisopropylbiphenyl; DIEA stands for N, N-diisopropylethylamine.

The compounds are prepared by hand orThe software names, and the commercial compounds are under the supplier catalog name.

Detailed Description

The present invention is described in detail below by way of examples, but is not meant to be limited to any of the disadvantages of the present invention. Having described the invention in detail and having disclosed specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof.

Example 1

Step 1: compound 1-1 (11.30g.80.14mmol) was added to anhydrous THF (120.00 mL), iPrMgCl (2M, 40.07mL) was added dropwise at-78 deg.C, and stirred at 15 deg.C under nitrogen for 16 hours. After the reaction is finished, saturated NH is added ₄ Cl (50 mL), extracted and concentrated, and the residue was purified by silica gel column chromatography to give compound 1-2. ¹ HNMR(400MHz，CDCl ₃ )δ3.57(t，J＝6.0Hz，2H)，2.59～2.66(m，3H)，2.02～2.07(m，2H)，1.11(d，J＝6.8Hz，6H).

Step 2: compound 1-2 (3.70g, 24.89mmol) and 2-chloro-4-bromopyridine (4.79g, 24.89mmol) were added to anhydrous THF (50.00 mL), n-BuLi (2.5M, 9.96mL) was added dropwise at-78 deg.C, and stirred at 20 deg.C for 18 hours under nitrogen. After the reaction is finished, adding saturated NH ₄ And Cl, extracting and concentrating, and purifying the residue by silica gel column chromatography to obtain the compound 1-3. ¹ HNMR(400MHz，CDCl ₃ )δ8.31(d，J＝6.8Hz，1H)，7.33(d，J＝1.6Hz，1H)，7.15～7.19(m，1H)，3.92～3.98(m，1H)，3.75～3.81(m，1H)，2.08～2.17(m，2H)，1.89～2.01(m，2H)，1.60～1.72(m，2H)，0.78(d，J＝6.8Hz，3H)，0.72(d，J＝6.8Hz，3H).

And step 3: compound 1-3 (1.60g, 7.09mmol), KOH (795.64mg, 14.18mmol) was added to dioxane (15.00 mL)/H ₂ O (5.00 mL) in a mixed solvent, followed by addition of Pd ₂ (dba) ₃ (324.62mg, 354.50. Mu. Mol) and tBuXPhOS (301.07mg, 709.00. Mu. Mol), under nitrogen, stirring at 115 ℃ for 18 hours. After the reaction, the mixture was diluted with water and extracted with ethyl acetate, and the extract was dried with brine and anhydrous sodium sulfate and concentrated. The residue was purified by silica gel column chromatography to obtain compounds 1 to 4.MS ESI calculated value C ₁₂ H ₁₆ ClNO[M+H] ⁺ 208, found 208.

And 4, step 4: compounds 1-4 (200.00mg, 964.92. Mu. Mol) were added to anhydrous DMF (50.00 mL), naH (60%, 57.90mg, 1.45mmol) was added at 0 deg.C, stirring was carried out for 0.5 h at 25 deg.C, followed by methyl 4-bromomethylbenzoate (221.04mg, 964.92. Mu. Mol) at 0 deg.C and stirring was carried out for 0.5 h at 25 deg.C. And after the reaction is finished, adding 20mL of water, filtering, drying filter residues, and performing column chromatography purification to obtain the compound 1-5.MS ESI calculated value C ₂₁ H ₂₅ NO ₄ [M+H] ⁺ 356, found 356.

And 5: compounds 1-5 (100mg, 281.35. Mu. Mol) were added to DCM/H ₂ O (15.00 mL) in a mixed solvent, 2M NaOH (2 mL) at 0 deg.C, and then 50% NH ₂ OH (2 mL), stirred at 25 ℃ for 0.5 h. Separation by preparative HPLC afforded the desired compounds 1-6. ¹ H NMR(400MHz，MeOD)δ7.74(d，J＝8.0Hz，2H)，7.68(d，J＝6.8Hz，1H)，7.40(d，J＝8.0Hz，2H)，6.61(d，J＝0.8Hz，1H)，6.46(dd，J＝6.8Hz&0.8hz, 1h), 5.25 (s, 2H), 3.91 to 3.97 (m, 1H), 3.79 to 3.84 (m, 1H), 2.10 to 2.19 (m, 3H), 2.01 to 2.05 (m, 1H), 1.79 to 1.94 (m, 1H), 0.95 (d, J =7.2hz, 3h), 0.83 (d, J =7.2hz, 3h), MS ESI calculated value C ₂₀ H ₂₄ N ₂ O ₄ [M+H] ⁺ 357, found value 357.

Example 2

Step 1: to tetrahydrofuran (100.00 mL) of the compound 2-1 (10.00g, 51.96mmol, 1.00eq) and 5-chloropent-2-one (6.27g, 51.96mmol, 1.00eq) was added dropwise n-butyllithium (2.5M, 24.94mL, 1.20eq) under a nitrogen atmosphere at-78 ℃ to 15 ℃ and stirred for 5 hours under a nitrogen atmosphere. The reaction mixture was concentrated, dispersed in water (100 mL) and ethyl acetate (100 mL), the combined organic phases were extracted, washed with saturated brine (100mL. Multidot.3), and washed with Na ₂ SO ₄ Drying, concentrating, and purifying with silica gel column to obtain compound 2-2.MS ESI calculated value C ₁₀ H ₁₂ ClNO[M+H] ⁺ 198, found 198.

Step 2: to a solution of Compound 2-2 (3.00g, 15.18mmol, 1.00eq) and potassium hydroxide (1.28g, 22.77mmol, 1.50eq) in dioxane (15.00 mL) and water (3.00 mL) under a nitrogen atmosphere was added Pd ₂ (dba) ₃ (1.39g, 1.52mmol, 0.10eq) and t-BuXphos (1.29g, 3.04mmol, 0.20eq). The mixture was purged with nitrogen 3 times, and then stirred at 110 ℃ for 12 hours under nitrogen atmosphere. The reaction solution was filtered by suction, poured into water, extracted with dichloromethane/methanol (10: 1, 20mL × 3), washed with saturated brine, dried over sodium sulfate, concentrated, and purified by silica gel column to obtain compound 2-3.MS ESI calculated value C ₁₀ H ₁₃ NO[M+H] ⁺ 180, found 180.

And 3, step 3: the compound 2-3 (230.00mg, 1.28mmol, 1.00eq), methyl 4- (bromomethyl) benzoate (293.97mg, 1.28mmol, 1.00eq), cesium carbonate (627.21mg, 1.93m m)mol,1.50 eq), potassium iodide (10.65mg, 64.17. Mu. Mol,0.05 eq) in tetrahydrofuran (20.00 mL) was replaced with nitrogen gas 3 times, followed by stirring at 66 ℃ under nitrogen atmosphere for 12 hours. The reaction solution was concentrated, then dissolved in water, extracted with ethyl acetate, washed with saturated brine, dried over sodium sulfate, and concentrated to obtain compound 2-4.MS ESI calculated value C ₁₉ H ₂₁ NO ₄ [M+H] ⁺ 328, found 328.

And 4, step 4: to compound 2-4 (300.00mg, 916.39. Mu. Mol,1.00 eq) in methanol (15.00 mL) was added dropwise aqueous sodium hydroxide (2M, 2.29mL, 5.00eq) at 0 ℃ under a nitrogen atmosphere. The reaction solution was stirred at 15 ℃ for 2.5 hours. The reaction mixture was concentrated, dissolved in water (15 mL), extracted with dichloromethane: methanol (10: 1, 15ml × 3), washed with saturated brine, dried over sodium sulfate, concentrated, and separated and purified by preparative HPLC to give compound 2-5. ¹ H NMR(400MHz，CDCl ₃ ) δ 7.40 (m, 2H), 7.19 (d, J =7.2, 1h), 6.87 (m, 2H), 6.72 (s, 1H), 6.27 (d, J =5.6hz, 2h), 4.01 (m, 2H), 3.86 (d, J =6.0, 2h), 2.46 (m, 2H), 2.16 (s, 2H), 1.96 (m, 2H), 1.65 (m, 4H), 1.32 (s, 4H). MS ESI calcd C ₁₈ H ₂₀ N ₂ O ₄ [M+H] ⁺ 329 found, value 329.

Example 3

Step 1: a cyclohexane solution of n-butyllithium (2.5M, 49.89mL, 1.20eq) was added dropwise to a toluene (30.00 mL) solution of 4-bromo-2-chloro-pyridine (20.00g, 103.93mmol, 1.00eq) at-65 ℃ to 75 ℃ and stirred at 65 ℃ to 75 ℃ for 1 hour, and then a toluene (100.00 mL) solution of the compound 3-1 (20.85g, 103.93mmol,17.09mL, 1.00eq) was added to the reaction system and stirred at 25 ℃ for 2 hours. To the reaction solution was added a saturated aqueous ammonium chloride solution, extracted with ethyl acetate, then dried over anhydrous sodium sulfate, filtered, and concentrated. After concentration, the mixture was purified by a silica gel column to obtain compound 3-2. ¹ H NMR(400MHz，CDCl ₃ )δ8.29(d，J＝5.3Hz，1H)，7.43-7.37(m，3H)7.24 (dd, J =1.6,5.2hz, 1h), 7.06-7.00 (m, 2H), 4.13-4.00 (m, 2H), 2.64-2.56 (m, 1H), 2.46 (m, 1H), 2.04-1.91 (m, 2H). MS ESI calculated value C ₁₅ H ₁₃ ClFNO[M+H]+278, found 278.

Step 2: a reaction solution of compound 3-2 (8.50g, 30.61mmol, 1.00eq), potassium hydroxide (3.44g, 61.22mmol, 2.00eq), tris (dibenzylideneacetone) dipalladium (5.61g, 6.12mmol, 0.20eq), 2-di-t-butylphosphine-2 ',4',6' -triisopropylbiphenyl (2.60g, 6.12mmol, 0.20eq) dioxane (2.00 mL) and water (1.00 mL) was heated to 100 ℃ and stirred for 2 hours. 1M dilute hydrochloric acid is added dropwise to the reaction liquid to adjust the pH value to 7, water is added, ethyl acetate is used for extraction, anhydrous sodium sulfate is used for drying, filtration and concentration are carried out to obtain a crude product. Pulping and purifying to obtain the compound 3-3. ¹ H NMR(400MHz，CDCl ₃ ) δ 12.84 (brs, 1H), 7.40 (dd, J =5.3,8.7hz, 2h), 7.25 (d, J =6.8hz, 1h), 7.01 (t, J =8.8hz, 2h), 6.69 (s, 1H), 6.29 (dd, J =1.4,6.8hz, 1h), 4.12-3.98 (m, 2H), 2.48 (t, J =7.2hz, 2h), 1.98 (m, 2H), 1.25 (dd, J =6.9,9.5hz, 2h). MS ESI calculated value C ₁₅ H ₁₄ FNO ₂ [M+H] ⁺ 260, measured value 260.

And 3, step 3: to a solution of compound 3-3 (7.00g, 27.00mmol, 1.00eq) in N, N-dimethylformamide (70.00 mL) was added sodium hydride (1.08g, 27.00mmol, 60% purity, 1.00 eq) at 0-5 ℃ and stirred at 0-5 ℃ for 10 minutes, then to the reaction system was added a solution of methyl 4-bromomethylbenzoate (6.18g, 27.00mmol, 1.00eq) in N, N-dimethylformamide (20.00 mL) and stirred at 10-25 ℃ for 2 minutes. Saturated aqueous ammonium chloride solution was added to the reaction mixture, and anhydrous sodium sulfate was extracted with ethyl acetate, dried, and concentrated. After concentration, compounds 3-4 were obtained and used directly in the next reaction.

And 4, step 4: to a mixture of compound 3 to 4 (11.00g, 27.00mmol, 1.00eq), aqueous hydroxylamine solution (100.00mL, 50% purity), methanol (20.00 mL) and methylene chloride (10.00 mL) were added aqueous sodium hydroxide solution (2M, 27.00mL, 2.00eq) and aqueous hydroxylamine solution (1.00mL, 50% purity), the mixture was heated to 40 to 50 ℃ and stirred for 1 hour. Concentrating the reaction solution, adjusting the pH to 7 with 1M hydrochloric acid, and extracting with dichloromethaneTaking, concentrating, and purifying by reversed phase preparative HPLC to obtain the compound 3-5. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.17 (s, 1H), 9.04 (br s, 1H), 7.74-7.64 (m, 3H), 7.55-7.45 (m, 2H), 7.32 (d, J =8.3hz, 2h), 7.14 (t, J =8.9hz, 2h), 6.50 (d, J =1.9hz, 1h), 6.25 (dd, J =2.0,7.2hz, 1h), 6.29-6.22 (m, 1H), 5.05 (s, 2H), 3.98-3.83 (m, 2H), 2.48-2.41 (m, 2H), 1.85 (m, 2H). MS ESI calculated value C ₂₃ H ₂₁ FN ₂ O ₄ [M+H] ⁺ 409, measured value 409.

Example 4

Step 1: to a solution of compound 4-1 (1.50g, 5.79mmol) in dimethylformamide (20.00 mL) was added sodium hydride (347.13mg, 8.69mmol,60% purity), and after stirring at 0 ℃ for 30 minutes, methyl 4-bromomethylbenzoate (1.99g, 8.69mmol) was added, warmed to room temperature, and stirred for 2 hours. Quenched with saturated aqueous ammonium chloride (15 mL) at 0 ℃ and diluted with water (20 mL), extracted with ethyl acetate EtOAc (20ml × 4). The combined organic phases were washed with water (20mL × 2) and saturated brine (20mL × 1), dried, filtered, concentrated, and purified by silica gel column chromatography (PE/EtOAc =3/1, 1: 1 to 0/1) to give a crude mixture. The crude mixture was resolved by chiral resolution (chiral column model: OJ (250mm. About.30mm, 10 μm), mobile phase: A: CO 2B: 0.05% diethylamine/EtOH, etOH (0.05% diethylamine) and supercritical fluid CO2 from 5% to 40%, flow rate 60 mL/min) to give compound 4-2 and compound 5-1, retention time 2.226min,2.835min, respectively. Preparation of Compound 4-2 ¹ H NMR(400MHz，DMSO-d ₆ ) δ =7.95-7.88 (m, 2H), 7.71 (d, J =7.2hz, 1h), 7.54-7.46 (m, 2H), 7.38 (d, J =8.4hz, 2h), 7.19-7.09 (m, 2H), 6.51 (d, J =2.0hz, 1h), 6.27 (dd, J =2.0,7.2hz, 1h), 5.09 (s, 2H), 3.98-3.85 (m, 2H), 3.83 (s, 3H), 2.49-2.41 (m, 2H), 1.91-1.75 (m, 2H), MS ESI calculated value C ₂₄ H ₂₂ FNO ₄ [M+H] ⁺ 407.43, found 408.1.

Step 2: to a solution of compound 4-2 (700.00mg, 1.72mmol) in methanol (5.00 mL) was added sodium hydroxide (103.20mg, 2.58mmol) and hydroxylamine (5.00mL, 50% aqueous solution), and the mixture was stirred at room temperature for 30 minutes, after concentration, the pH was adjusted to 7 with dilute hydrochloric acid (1M), and the mixture was subjected to preparative HPLC (0.225% FA) on the reverse phase to isolate and purify compound 4-3. ¹ H NMR (400mhz, dmso-d 6) δ =11.49-10.50 (br s, 1H), 9.04 (br s, 1H), 7.69 (m, 3H), 7.49 (m, 2H), 7.33 (m, 2H), 7.14 (m, 2H), 6.50 (m, 1H), 6.25 (m, 1H), 5.05 (s, 2H), 3.90 (m, 2H), 2.45 (m, 2H), 1.85 (m, 2H). MS ESI calculated value C ₂₃ H ₂₁ N ₂ O ₄ F[M+H] ⁺ 408.42, found 408.9.

Example 5

Step 1: to a solution of compound 5-1 (900.00mg, 2.21mmol) in methanol (5.00 mL) were added sodium hydroxide (132.60mg, 3.32mmol) and hydroxylamine (5.00mL, 50% aqueous solution), and the mixture was stirred at room temperature for 30 minutes. Concentrating, adjusting pH to 7 with dilute hydrochloric acid (1M), extracting with ethyl acetate (50mL. X.4), combining the organic phases and concentrating, purifying by preparative reverse phase HPLC ((0.225% FA) to obtain compound 5-2. ¹ H NMR(400MHz，DMSO-d ₆ ) Δ 12.07-10.09 (br s, 1H), 9.08 (br s, 1H), 7.68 (M, 3H), 7.56-7.44 (M, 2H), 7.32 (M, 2H), 7.14 (M, 2H), 6.50 (M, 1H), 6.25 (M, 1H), 5.01 (s, 2H), 4.09-3.74 (M, 2H), 2.45 (M, 2H), 1.94-1.76 (M, 2H), MS ESI calculated value C23H21FN2O4[ M + H ] M] ⁺ 408.42, found 408.8.

Example 6

Step 1: after 4-chlorobutyryl chloride (500.00mg, 3.55mmol, 396.83. Mu.L, 0.50 eq) was added to a mixture of compound 6-1 (809.15mg, 7.09mmol, 697.54. Mu.L, 1.00 eq) and aluminum trichloride (633.60mg, 4.75mmol, 259.67. Mu.L, 0.67 eq) at 5 ℃ and stirred at 25 ℃ for 3 hours, the reaction mixture was added to ice water, stirred, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 6-2. The liquid was used directly in the next reaction.

And 2, step: to a solution of 4-bromo-2-chloro-pyridine (527.29mg, 2.74mmol, 1.00eq) in toluene (5.00 mL) at-68 ℃ was added a solution of n-butyllithium in cyclohexane (2.5M, 1.21mL, 1.10eq), and the mixture was stirred at 68 ℃ for 10 minutes, followed by addition of a solution of compound 6-2 (600.00mg, 2.74mmol, 1.00eq) in toluene (5.00 mL) to the reaction system, followed by stirring at-68 ℃ for 1 hour and at 25 ℃ for 14 hours. The reaction mixture was added with saturated aqueous ammonium chloride solution and water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. After concentration, purification by TLC plate gave compound 6-3.MS ESI calculated value C ₁₅ H ₁₂ F ₂ ONCl[M+H] ⁺ 296, found 296.

And step 3: to a solution of compound 6-3 (87.00mg, 294.21. Mu. Mol,1.00 eq) in dioxane (2.00 mL) and water (1.00 mL) were added potassium hydroxide (33.02mg, 588.42. Mu. Mol,2.00 eq), tris (dibenzylideneacetone) dipalladium (26.94mg, 29.42. Mu. Mol,0.10 eq), 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (12.49mg, 29.42. Mu. Mol,0.10 eq), and stirred at 80 ℃ for 3 hours. Water was added to the reaction mixture, which was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. After concentration, purification on TLC plates afforded compound 6-4.MS ESI calculated value C ₁₅ H ₁₃ F ₂ O ₂ N[M+H] ⁺ 278, found in 278.

And 4, step 4: to a solution of the compound 6-4 (60.00mg, 216.40. Mu. Mol,1.00 eq) in N, N-dimethylformamide (1.00 mL) at 0 ℃ was added sodium hydride (17.31mg, 432.80. Mu. Mol, purity 60%,2.00 eq) and then methyl 4-bromomethylbenzoate (49.57mg, 216.40. Mu. Mol,1.00 eq) in N, N-dimethylformamide (1.00 mL) was added to the reaction systemThe solution was stirred at 25 ℃ for 2 hours. Water was added to the reaction mixture, which was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. After concentration, purification by TLC plate afforded Compound 6-5.MS ESI calculated value C ₂₄ H ₂₁ NO ₄ F ₂ [M+H] ⁺ 426, found value 426.

And 5: to a solution of compound 6-5 (32.00mg, 75.22. Mu. Mol,1.00 eq) in methanol (2.00 mL) was added an aqueous sodium hydroxide solution (6M, 50.00. Mu.L, 3.99 eq) and an aqueous hydroxylamine solution (1.00mL, 50% aqueous solution), and the mixture was stirred at 25 ℃ for 17 hours. After the reaction solution is concentrated, the compound 6-6 is obtained by reversed phase preparative column purification. ¹ H NMR (400mhz, meod) δ ppm 1.89-2.08 (m, 2H) 2.48-2.74 (m, 2H) 3.93-4.11 (m, 2H) 5.20 (s, 2H) 6.46 (dd, J =7.09,1.82hz, 1h) 6.66 (s, 1H) 6.88-7.03 (m, 2H) 7.37 (d, J =8.16hz, 2h) 7.63 (d, J =7.15hz, 1h) 7.67-7.72 (m, 2H) 7.72-7.75 (m, 1H). MS ESI calculated value C ₂₃ H ₂₀ N ₂ O ₄ F ₂ [M+H] ⁺ 427, found 427.

Example 7

Step 1: to a solution of compound 7-1 (70.00mg, 269.99. Mu. Mol) in dimethylformamide (5.00 mL) was added sodium hydride (21.60mg, 539.98. Mu. Mol,60% purity), stirred at 0 ℃ for 30 minutes, and to the mixture was added 6-bromomethyl-3-carboxylic acid methyl ester pyridine (124.23mg, 539.98. Mu. Mol), and stirred at 0 ℃ for 30 minutes. The temperature was raised to room temperature and stirring was continued for 30 minutes. The mixture was extracted with saturated aqueous ammonium chloride (1 mL), water (15 mL), ethyl acetate (10 mL × 4). The combined organic phases were washed with water (10 mL × 3) and saturated brine (10 mL), dried, filtered and concentrated. Plate separation (PE/EA = 1/1) yielded compound 7-2.MS ESI calculated value C ₂₃ H ₂₁ FN ₂ O ₄ [M+H] ⁺ 409, measured value 409.

And 2, step: to a solution of Compound 7-2 (70.00mg, 161.11. Mu. Mol) in methanol (5.00 mL) was added hydroxylamine (1.00mL, 50% aqueous solution) and hydrogen oxygenSodium hydroxide NaOH (6.44mg, 161.11. Mu. Mol), and stirred at room temperature for 30 minutes. The mixture was adjusted to pH 7 with dilute hydrochloric acid (1M) and purified to give compound 7-3. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 8.77 (d, J =1.8hz, 1h), 8.05 (dd, J =2.4,8.2hz, 1h), 7.69 (d, J =7.2hz, 1h), 7.57-7.48 (m, 2H), 7.30 (d, J =8.2hz, 1h), 7.21-7.11 (m, 2H), 6.48 (d, J =1.8hz, 1h), 6.28 (dd, J =2.0,7.2hz, 1h), 5.14 (s, 2H), 4.01-3.83 (m, 2H), 2.49-2.44 (m, 2H), 1.92-1.79 (m, 2H), MS ESI calculated value C ₂₂ H ₂₀ FN ₃ O ₄ [M+H] ⁺ 410, found value 410.

Example 8

Step 1: to a solution of compound 8-1 (70.00mg, 269.99. Mu. Mol) in dimethylformamide (5.00 mL) was added sodium hydride (21.60mg, 539.98. Mu. Mol, purity 60%), stirred at 0 ℃ for 30 minutes, then 2-bromomethyl 5-carboxylic acid methyl ester pyrimidine (132.33mg, 539.98. Mu. Mol), stirred at 0 ℃ for 30 minutes, warmed to room temperature and stirred for 30 minutes. The mixture was quenched with saturated ammonium chloride (1 mL), water (15 mL) was added, and extracted with ethyl acetate (10 mL 4). The combined organic phases were washed with water (5 mL × 3) and saturated brine (10 mL), dried, filtered, concentrated, and purified by preparative thin layer chromatography (PE/EA = 1/1) to give compound 8-2.MS ESI calculated value C22H20FN3O4[ M + H ]] ⁺ 424, found value 424.

Step 2: to a solution of Compound 8-2 (70.00mg, 160.72. Mu. Mol) in methanol (5.00 mL) were added hydroxylamine (1.00mL, 50% aqueous solution) and sodium hydroxide NaOH (6.43mg, 160.72. Mu. Mol), and the mixture was stirred at room temperature for 30 minutes. The mixture was adjusted to pH 7 with dilute hydrochloric acid (1M) and purified to give compound 8-3. ¹ H NMR (400MHz, DMSO-d 6) delta 8.97 (s, 2H), 7.69 (d, J =7.2Hz, 1H), 7.59-7.51 (m, 2H), 7.20-7.13 (m, 2H), 6.47 (d, J =2.0Hz, 1H), 6.31 (dd, J =2.0,7.2Hz, 1H), 5.26 (s, 2H), 4.00-3.87 (m, 2H), 2.53 (m, 2H), 1.93-1.82 (m, 2H), MS ESI calculated value C ₂₁ H ₁₉ FN ₄ O ₄ [M+H] ⁺ 411, found 411.

Example 9

Step 1: to a solution of compound 9-1 (70.00mg, 269.99. Mu. Mol) in dimethylformamide (5.00 mL) was added sodium hydride (21.60mg, 539.98. Mu. Mol, purity 60%), stirred at 0 ℃ for 30 minutes, followed by addition of methyl 2-carboxylate-5-bromomethylpyridine (124.23mg, 539.98. Mu. Mol), stirring was continued at 0 ℃ for 30 minutes, and after warming to room temperature, stirring was continued for 30 minutes. The mixture was quenched with saturated ammonium chloride (1ml × 1), water (15 mL) was added, and extracted with ethyl acetate (10ml × 3). The combined organic phases were washed with water (10 mL × 2) and saturated brine (10 mL), dried, filtered, concentrated, and purified by preparative TLC (PE/EA = 1/1) to give compound 9-2.MS ESI calculated value C ₂₃ H ₂₁ FN ₂ O ₄ [M+H] ⁺ 409, measured value 409.

Step 2: to a solution of compound 9-2 (50.00 mg, crude) in methanol (5.00 mL) was added hydroxylamine (1.00mL, 50% aqueous solution) and sodium hydroxide NaOH (3.06mg, 76.39. Mu. Mol), and the mixture was stirred at room temperature for 30 minutes. The mixture was adjusted to pH 7 with dilute hydrochloric acid (1M) and purified to give compound 9-3. ¹ H NMR(400MHz，DMSO-d ₆ ) Δ 8.53 (s, 1H), 7.97-7.87 (m, 1H), 7.86-7.72 (m, 2H), 7.50 (m, 2H), 7.14 (m, 2H), 6.51 (m, 1H), 6.28 (m, 1H), 5.10 (s, 2H), 4.01-3.81 (m, 2H), 2.48-2.42 (m, 2H), 1.92-1.76 (m, 2H). MS ESI calculated value C ₂₂ H ₂₀ FN ₃ O ₄ [M+H] ⁺ 410, found value 410.

Example 10

Step 1: to a mixture of compound 10-1 (4.68g, 35.46mmol,3.66mL, 2.00eq) and aluminum trichloride (3.14g, 23.58mmol,1.29mL, 1.33eq) was added dropwise 4-chlorobutyryl chloride (2.50g, 17.73mmol,1.98mL, 1.00eq) and stirred at 30 ℃ for 6 hours, and then the reaction mixture was added to ice water, stirred, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. After concentration, the mixture was purified by a silica gel column to obtain compound 10-2. The liquid was used directly in the next reaction.

Step 2: to a solution of 4-bromo-2-chloro-pyridine (488.80mg, 2.54mmol, 1.00eq) in toluene (3.00 mL) at-68 ℃ was added a cyclohexane solution of n-butyllithium (2.5M, 1.12mL, 1.10eq), and then to the reaction system was added a toluene (1.00 mL) solution of compound 10-2 (600.00mg, 2.54mmol, 1.00eq), and the mixture was stirred at 25 ℃ for 6 hours. To the reaction mixture were added a saturated aqueous ammonium chloride solution and water, followed by extraction with ethyl acetate, washing with saturated brine, drying over anhydrous sodium sulfate, and concentration. After concentration, the mixture was purified by a silica gel column to obtain compound 10-3.MS ESI calculated value C ₁₅ H ₁₁ F ₃ ONCl[M+H] ⁺ 314, measured value 314.

And step 3: to a solution of compound 10-3 (410.00mg, 1.31mmol, 1.00eq) in dioxane (5.00 mL) and water (1.00 mL) was added potassium hydroxide (146.67mg, 2.61mmol, 2.00eq), tris (dibenzylideneacetone) dipalladium (119.96mg, 131.00. Mu. Mol,0.10 eq), 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (55.50mg, 130.70. Mu. Mol,0.10 eq), and stirred at 80 ℃ for 4 hours. To the reaction solution was added a saturated aqueous ammonium chloride solution, water, and extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Concentrating, purifying with silica gel column to obtain compound 10-4.

And 4, step 4: a solution of compound 10-4 (100.00mg, 338.68. Mu. Mol,1.00 eq), methyl 4-bromomethylbenzoate (155.16mg, 677.36. Mu. Mol,2.00 eq), cesium carbonate (220.70mg, 677.37. Mu. Mol,2.00 eq) in acetonitrile (2.00 mL) was warmed to 80 ℃ and stirred for 2 hours. Water was added to the reaction mixture, which was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. After concentration, purification by preparative thin layer chromatography gave compound 10-5.

And 5: to a solution of compound 10-5 (40.00mg, 90.21. Mu. Mol,1.00 eq) in methanol (2.00 mL) was added sodium hydroxide (3.61mg, 90.21. Mu. Mol,1.00 eq) and an aqueous solution of hydroxylamine (1.00mL, 50% purity), and the mixture was stirred at 25 ℃ for 17 hours. After the reaction solution is concentrated, the compound 10-6 is obtained by reversed phase preparative column purification. ¹ H NMR (400mhz, meod) δ ppm 1.75-2.11 (M, 2H) 2.53-2.84 (M, 2H) 3.90-4.20 (M, 2H) 5.41 (s, 2H) 6.79-6.88 (M, 2H) 6.92 (s, 1H) 6.97 (dd, J =5.40,1.25hz, 1h) 7.54 (d, J =8.28hz, 2h) 7.76 (d, J =8.28hz, 2h) 8.05 (d, J =5.40hz, 1h), MS ESI calcd 23H19N2O4F3[ M + H4F 3 ] (M + H) ("J =5.40hz, 1h)] ⁺ 445, found 445.

Example 11

Step 1: at N ₂ To a solution of 4-bromo-2-chloro-3-fluoropyridine (4.00g, 19.01mmol, 1.00eq) in toluene (20.00 mL) at-70 ℃ was added n-BuLi (2.5M, 9.12mL, 1.20eq) dropwise, and after completion of the addition, stirring was continued at that temperature for 30 minutes, and then compound 11-1 (3.81g, 19.01mmol, 819.67. Mu.L, 1.00 eq) was added to the suspension. The temperature of the reaction system was gradually raised to 40 ℃ and stirred for 12 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc (40mL x3), and the combined extracts were washed with brine (50 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to silica gel column chromatography (PE/EtOAc = 10/1) to give compound 11-2.MS ESI calculated value C ₁₅ H ₁₂ ClF ₂ NO[M+H] ⁺ 296, found 296.

Step 2: at N ₂ Protection, compound 11-2 (3.86g, 13.05mmol, 1.00eq), t-BuXPhOS (554.16mg, 1.31mmol, 0.10eq), pd ₂ (dba) ₃ (1.20g, 1.31mmol, 0.10eq) and KOH (1.46g, 26.10mmol, 2.00eq) and H ₂ The O (20 mL) mixture was heated to 100 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc/MeOH (10/1, 30mL x3), and the combined extracts were washed withSaline (40 mL) was washed with brine, followed by anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. Pulping the obtained solid by PE (20 mL), filtering and drying to obtain the compound 11-3.MS ESI calculated value C ₁₅ H ₁₃ F ₂ NO ₂ [M+H] ⁺ 278, found 278.

And step 3: at N ₂ Protection, naH (553.97mg, 13.85mmol, 60% purity, 1.20 eq) was added to a solution of compound 11-3 (3.20g, 11.54mmol, 1.00eq) in DMF (60 mL) at 0 ℃ and stirred at that temperature for 20 minutes, then methyl 4-bromomethylbenzoate (2.64g, 11.54mmol, 1.10eq) was added to the suspension and the system temperature was raised to 20 ℃ and stirring was continued for 1 hour. The reaction solution was poured into H ₂ O (250 mL) and extracted with EtOAc/MeOH (10/1, 50mL x3), and the combined extracts were washed with brine (100 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to silica gel column chromatography (PE/EtOAc = 3/1) to obtain compound 11-4.MS ESI calculated value C ₂₄ H ₂₁ F ₂ NO ₄ [M+H] ⁺ 426, found value 426.

And 4, step 4: to a solution of compounds 11-4 (2.00g, 4.70mmol, 1.00eq) in MeOH (20.0 mL) and DCM (10.0 mL) was added NH in succession dropwise at 0 deg.C ₂ OH.H ₂ O (20.0 mL,50% aqueous solution) and NaOH (2M, 20.0 mL) aqueous solution, and the reaction was continued to stir at 0 ℃ for 1 hour. Concentrating under reduced pressure to remove most solvent, cooling the residual solution to 0 deg.C, adjusting pH =7-8 with concentrated hydrochloric acid, and post-treating to obtain compound 11-5.1H NMR (400MHz, DMSO-d) ₆ ) δ 11.18 (s, 1H), 9.90-9.90 (m, 1H), 9.04 (brs, 1H), 7.98 (d, J =6.78hz, 1H), 7.70 (d, J =8.03hz, 2h), 7.44 (dd, J =8.41,5.65hz, 2h), 7.37 (d, J =8.03hz, 2h), 7.15 (t, J =8.91hz, 2h), 6.60 (d, J =7.53hz, 1H), 4.90-5.15 (m, 2H), 3.80-4.05 (m, 2H), 2.60-2.75 (m, 1H), 2.38-2.44 (m, 1H), 1.93 (q, J =7.09hz, 2h), MS ESI calculated value C ₂₃ H ₂₀ F ₂ N ₂ O ₄ [M+H]+427, found 427.

Example 12

Step 1: in N ₂ To a solution of 4-bromo-2-chloropyridine (515.62mg, 2.68mmol, 1.00eq) in toluene (3.00 mL) at-70 ℃ was added n-BuLi (2.5M, 1.29mL, 1.20eq) dropwise, and after completion of the addition, stirring was continued at that temperature for 20 minutes, and then compound 12-1 (500.00mg, 2.68mmol, 1.00eq) was added to the above suspension. The reaction temperature was gradually raised to 25 ℃ and stirred for 6 hours. Toluene was removed by concentration under reduced pressure, the remaining solid was dissolved in DMF (3.00 mL), naH (160.76mg, 4.02mmol, purity 60%,1.50 eq) was added at 25 ℃ and the system was heated to 60 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc (30mL x3), and the combined extracts were washed with brine (50 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to silica gel column chromatography (PE/EtOAc = 10/1) to give compound 12-2.MS ESI calculated value C ₁₄ H ₁₁ ClFNO[M+H] ⁺ 264, found 264.

Step 2: at N ₂ Protection, compound 12-2 (320.00mg, 1.21mmol, 1.00eq), t-BuXPhOS (102.76mg, 242.00. Mu. Mol,0.20 eq), pd ₂ (dba) ₃ (221.60mg, 242.00. Mu. Mol,0.20 eq) and KOH (135.79mg, 2.42mmol, 2.00eq) in dioxane (10.00 mL) and H ₂ The O (3.00 mL) mixture was heated to 100 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL x3), and the combined extracts were washed with brine (60 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to preparative TLC (PE/EtOAc = 3/1) to afford compound 12-3.MS ESI calculated value C ₁₄ H ₁₂ FNO ₂ [M+H] ⁺ 246, found 246.

And step 3: in N ₂ Protection, compound 12-3 (290.00mg, 1.18mmol, 1.00eq) in DMF (5.00 mL) at 0 ℃ was added NaH (56.80 mg,1.42mmol, 60% purity, 1.20 eq) and stirred at this temperature for 20 minutes, then methyl 4-bromomethylbenzoate (297.33mg, 1.30mmol,1.10 eq) was added to the suspension and the system temperature was raised to 20 ℃ and stirring continued for 2 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc/MeOH (10/1, 30mL x3), and the combined extracts were washed with brine (50 mL) and then anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was isolated and purified by preparative TLC (PE/EtOAc = 1/1) to afford compound 12-4.MS ESI calculated value C ₂₃ H ₂₀ FNO ₄ [M+H] ⁺ 394, found 394.

And 4, step 4: to a solution of compounds 12-4 (150.00mg, 381.28. Mu. Mol,1.00 eq) in DCM (2.00 mL) and MeOH (2.00 mL) was added dropwise NH in that order at 0 deg.C ₂ OH.H ₂ O (150.00. Mu.L of 50% aqueous solution) and NaOH (2M, 150.00. Mu.L) aqueous solution, and the reaction was then continued to stir at 0 ℃ for 2 hours. Concentrating under reduced pressure to remove most of the solvent, and subjecting the remaining solution to preparative HPLC (0.1% NH) ₄ OH) is separated and purified to obtain the compound 12-5. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 10.99 (brs, 1H), 8.67-9.79 (brs, 1H), 7.77 (d, J =7.28hz, 1h), 7.69 (d, J =8.03hz, 2h), 7.46 (dd, J =8.66,5.40hz, 2h), 7.34 (d, J =8.03hz, 2h), 7.20 (t, J =8.91hz, 2h), 6.52 (d, J =1.51hz, 1h), 6.18 (dd, J =7.03,1.76hz, 1h), 5.08 (s, 2H), 4.39-4.58 (m, 2H), 3.01-3.19 (m, 2H), MS ESI calcd C ₂₂ H ₁₉ FN ₂ O ₄ [M+H]+395, measured value 395.

Example 13

Step 1: in N ₂ Protection n-BuLi (2.5M, 1.03mL, 1.20eq) was added dropwise to a toluene (8.00 mL) solution of 4-bromo-2-chloro-3-fluoropyridine (450.32mg, 2.14mmol, 1.00eq) at-70 ℃ and after completion of the addition, stirring was continued at that temperature for 20 minutes, and then Compound 13-1 (400.00mg, 2.14mmol, 1.00eq) was added to the above suspension. Gradually raising the temperature of the reaction system to 25 DEG CStirred for 6 hours. Toluene was removed by concentration under reduced pressure, the remaining solid was dissolved in DMF (8.00 mL), naH (128.40mg, 3.21mmol, purity 60%,1.50 eq) was added at 25 ℃ and the system was heated to 60 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc (30mL x3), and the combined extracts were washed with brine (50 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to silica gel column chromatography (PE/EtOAc = 10/1) to obtain compound 13-2.MSESI calculated value C ₁₄ H ₁₀ ClF ₂ NO[M+H] ⁺ 282, measured value 282.

Step 2: in N ₂ Protection, compound 13-2 (440.00mg, 1.56mmol, 1.00eq), t-BuXPhOS (132.49mg, 312.00. Mu. Mol,0.20 eq), pd ₂ (dba) ₃ (285.70mg, 312.00. Mu. Mol,0.20 eq) and KOH (175.06mg, 3.12mmol, 2.00eq) in dioxane (10.00 mL) and H ₂ A mixture of O (3.00 mL) was heated to 100 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (60 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to preparative TLC (PE/EtOAc = 3/1) to afford compound 13-3.MS ESI calculated value C ₁₄ H ₁₁ F ₂ NO ₂ [M+H] ⁺ 264, found 264.

And step 3: in N ₂ Protection, naH (69.29mg, 1.73mmol, 60% pure, 1.20 eq) was added to a solution of compound 13-3 (380.00mg, 1.44mmol, 1.00eq) in DMF (5.00 mL) at 0 ℃ and stirred at this temperature for 20 minutes, then methyl 4-bromomethylbenzoate (363.74mg, 1.59mmol, 1.10eq) was added to the suspension and the system temperature was raised to 20 ℃ and stirring continued for 2 hours. The reaction solution was poured into H ₂ O (80 mL) and extracted with EtOAc/MeOH (10/1, 30mL x3), and the combined extracts were washed with brine (50 mL) and then anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The obtained solid was isolated and purified by preparative TLC (PE/EtOAc = 1/1) to give compound 13-4.MS ESI calculated value C ₂₃ H ₁₉ F ₂ NO ₄ [M+H] ⁺ 412, measured value 412.

And 4, step 4: to a solution of compounds 13-4 (350.00mg, 850.75. Mu. Mol,1.00 eq) in DCM (3.00 mL) and MeOH (3.00 mL) was added dropwise NH in that order at 0 deg.C ₂ OH.H ₂ O (3.00mL, 50% aqueous solution) and NaOH (2M, 3.00mL) in water, and the reaction was allowed to continue stirring at 0 deg.C for 1 hour. Concentrating under reduced pressure to remove most of the solvent, and separating and purifying the remaining solution by preparative HPLC (0.1% TFA) to obtain Compound 13-5. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.19 (brs, 1H), 8.06 (d, J =6.52hz, 1h), 7.71 (d, J =8.28hz, 2h), 7.38-7.50 (m, 5H), 7.22 (t, J =8.91hz, 2h), 6.65 (d, J =7.28hz, 1h), 4.98-5.08 (m, 2H), 4.49-4.59 (m, 2H), 3.25-3.34 (m, 1H), 3.05-3.15 (m, 1H). MS ESI calcd C ESI ₂₂ H ₁₈ F ₂ N ₂ O ₄ [M+H]+413, found 413.

Example 14

Step 1: in N ₂ Protection n-BuLi (2.5M, 6.84mL, 1.20eq) was added dropwise to a toluene (80.00 mL) solution of 4-bromo-2-chloro-5-fluoropyridine (3.00g, 14.26mmol, 1.00eq) at-70 ℃ and after completion of the addition, stirring was continued at that temperature for 30 minutes, and then compound 14-1 (2.86g, 14.26mmol,2.34mL, 1.00eq) was added to the above suspension. The temperature of the reaction system was gradually raised to 40 ℃ and stirred for 12 hours. The reaction solution was poured into H ₂ O (300 mL) and extracted with EtOAc (100mL x3), and the combined extracts were washed with brine (100 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to silica gel column chromatography (PE/EtOAc = 10/1) to give compound 14-2.MS ESI calculated value C ₁₅ H ₁₂ ClF ₂ NO[M+H] ⁺ 296, found 296.

Step 2: in N ₂ Protection, compound 14-2 (300.00mg, 1.01mmol, 1.00eq), t-BuXPhOS(85.78mg，202.00μmol，0.20eq)，Pd ₂ (dba) ₃ (184.98mg, 202.00. Mu. Mol,0.20 eq) and KOH (113.34mg, 2.02mmol, 2.00eq) dioxane (10.00 mL) and H ₂ A mixture of O (3.00 mL) was heated to 100 ℃ and stirred for 2 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (60 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was subjected to preparative TLC (PE/EtOAc = 3/1) to afford compound 14-3.MS ESI calculated value C ₁₅ H ₁₃ F ₂ NO ₂ [M+H] ⁺ 278, found in 278.

And step 3: at N ₂ Protection, naH (34.62mg, 865.58. Mu. Mol,60% purity, 1.20 eq) was added to a solution of compound 14-3 (200.00mg, 721.32. Mu. Mol,1.00 eq) in DMF (5.00 mL) at 0 ℃ and stirred at this temperature for 20 minutes, then methyl 4-bromomethylbenzoate (165.23mg, 721.32. Mu. Mol,1.00 eq) was added to the suspension and the temperature of the system was raised to 20 ℃ and stirring was continued for 1 hour. The reaction solution was poured into H ₂ O (50 mL) and extracted with EtOAc/MeOH (10/1, 20mL of x3), and the combined extracts were washed with brine (30 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure. The obtained solid was isolated and purified by preparative TLC (PE/EtOAc = 1/1) to give compound 14-4.MS ESI calculated value C ₂₄ H ₂₁ F ₂ NO ₄ [M+H] ⁺ 426, found value 426.

And 4, step 4: to a solution of compound 14-4 (100.00mg, 235.06. Mu. Mol,1.00 eq) in DCM (2.00 mL) and MeOH (4.00 mL) at 0 ℃ is added NH dropwise in sequence ₂ OH.H ₂ O (2.00mL, 50% aqueous solution) and NaOH (2.00mL, 2M) in water, and the reaction was allowed to continue stirring at 0 ℃ for 1 hour. Concentrating under reduced pressure to remove most solvent, cooling the rest solution to 0 deg.C, adjusting pH =6-7 with concentrated hydrochloric acid, and post-treating to obtain compound 14-5. ¹ H NMR(400MHz，DMSO-d ₆ )δ11.17(brs，1H)，9.05(brs，1H)，8.61-9.43(m，1H)，7.64-7.73(m，3H)，7.46(dd，J＝8.5，5.5Hz，2H)，7.34(d，J＝8.0Hz，2H)，7.16(t，J＝8.9Hz，2H)，6.48(t，J＝6.9Hz，1H)，5.07-5.18(m，2H)，3.90(t，J＝7.2Hz，2H)，2.64-2.70 (m, 1H), 2.41-2.47 (m, 1H), 1.82-1.97 (m, 2H). MS ESI calculated value C ₂₃ H ₂₀ F ₂ N ₂ O ₄ [M+H]+427, found 427.

Example 15

Step 1: compound 15-1 (6.20g, 64.50mmol,6.08mL, 2.00eq) and aluminum trichloride (4.73g, 35.48mmol, 1.10eq) were charged into a 100-mL three-necked flask at 0 ℃ and 5-chloropentanoyl chloride (5.00g, 32.25mmol,4.17mL, 1.00eq) was added, and the mixture was warmed to room temperature and stirred for two hours. The mixture was quenched with ice water (20 mL), water (100 mL) was added, extracted with ethyl acetate (100ml × 2), the organic phases were combined, washed with saturated brine, dried over sodium sulfate, filtered and concentrated to give compound 15-2. ¹ H NMR(400MHz，CDCl ₃ )δ8.08-7.91(m，2H)，7.13(m，2H)，3.63-3.55(m，2H)，2.99(m，2H)，1.98-1.82(m，4H).

And 2, step: tert-butyllithium (2.5M, 13.32mL, 1.10eq) was slowly added dropwise to a toluene (100.00 mL) solution of 4-bromo-2-chloropyridine (5.83g, 30.28mmol, 1.00eq) under a nitrogen atmosphere at-78 ℃, and after stirring for ten minutes, compound 15-2 (6.50g, 30.28mmol, 1.00eq) was slowly added dropwise and stirring was continued for thirty minutes. After quenching with saturated ammonium chloride (20 mL), water (100 mL) was added, extraction was performed with ethyl acetate (100ml × 3), the combined organic phases were washed with saturated brine (100 mL), dried over sodium sulfate, filtered, concentrated, and column separated (silica gel column, petroleum ether/ethyl acetate = 10/1) to obtain compound 15-3.MS ESI calculated value C ₁₆ H ₁₆ Cl ₂ FNO[M+H] ⁺ 328.2, found 328.2.

And 3, step 3: to a solution of compound 15-3 (6.00g, 18.28mmol, 1.00eq) in N, N-dimethylformamide (30.00 mL) was added NaH (877.49mg, 21.94mmol,60% pure, 1.20 eq) under a nitrogen atmosphere at 0 ℃ and stirred for thirty minutes at 0-5 ℃. Quenching with saturated ammonium chloride (30 mL), adding water (100 mL), extracting with ethyl acetate (80mL. Multidot.3), and mixing the organic phases, extracting with water (100mL. Multidot.3), and saturatingWashed with brine (50ml × 1), dried over sodium sulfate, filtered and concentrated to give compound 15-4.MS ESI calculated value C ₁₆ H ₁₅ ClFNO[M+H] ⁺ 291.75, found 292.0.

And 4, step 4: under nitrogen protection, a mixed solution of 15-4 (1.00g, 3.43mmol, 1.00eq), potassium hydroxide (384.64mg, 6.86mmol, 2.00eq), tris (dibenzylideneacetone) dipalladium (627.74mg, 686.00. Mu. Mol,0.20 eq) and 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (291.10mg, 686.00. Mu. Mol,0.20 eq) were added to a mixed solution of water (5.00 mL) and 1, 4-dioxane (10.00 mL), and the mixture was heated to 90 ℃ and stirred for three hours. The mixture was concentrated, water (50 mL) was added, ethyl acetate (20ml × 3) was extracted, and the combined organic phase was washed with saturated brine (50 mL), dried over sodium sulfate, filtered, concentrated, and column-purified (silica gel column, petroleum ether/ethyl acetate =10/1 to 2/1, dichloromethane/methanol = 20/1) to obtain compound 15-5.MS ESI calculated value C ₁₆ H ₁₆ FNO ₂ [M+H] ⁺ 273.30, found 274.1.

And 5: to a solution of compounds 15-5 (150.00mg, 439.08. Mu. Mol,1.00 eq) in N, N-dimethylformamide (5.00 mL) was added sodium hydrogen (17.56mg, 439.08. Mu. Mol,60% purity, 1.00 eq) under nitrogen protection at 0 ℃, the mixture was stirred for ten minutes, then methyl 4-bromomethylbenzoate (125.72mg, 548.85. Mu. Mol,1.00 eq) was added, the mixture was allowed to warm to room temperature, and stirring was continued for thirty minutes. To the system was added saturated ammonium chloride (2 mL), water (15 mL) was added, extraction was performed with ethyl acetate (10 mL × 3), and the combined organic phases were washed with saturated brine (15 mL), dried over sodium sulfate, filtered, concentrated, and purified to obtain compound 15-6.MS ESI calculated value C ₂₅ H ₂₄ FNO ₄ [M+H] ⁺ 421.46, found 422.2.

Step 6: to a solution of compound 15-6 (145.00mg, 292.44. Mu. Mol,1.00 eq) in methanol (10.00 mL) were added an aqueous hydroxylamine solution (2.00mL, 50% aqueous solution) and sodium hydroxide (35.09mg, 877.32. Mu. Mol,3.00 eq), and the mixture was concentrated by stirring at room temperature for one hour, followed by preparative HPLC (0.225% FA) separation to give compound 15-7. ¹ H NMR(400MHz，DMSO-d ₆ )δ＝7.73-7.62(m，3H)，7.46(m，2H)，7.32(d，J＝8.2hz, 2h), 7.16 (t, J =8.8hz, 2h), 6.47 (d, J =1.6hz, 1h), 6.20 (dd, J =1.6,7.2hz, 1h), 5.04 (s, 2H), 3.71-3.53 (m, 2H), 2.34-2.07 (m, 2H), 1.65-1.46 (m, 4H), MS ESI calcd C ₂₄ H ₂₃ FN ₂ O ₄ [M+H] ⁺ 422.45, found 423.4.

Example 16

Step 1: under the protection of nitrogen at-78 ℃, n-butyllithium (2.5m, 88.63ml, 1.10eq) is slowly dropwise added into a toluene (250.00 mL) solution of the compound 16-1 (38.76g, 201.43mmol, 1.00eq), after stirring for thirty minutes, p-fluorobenzaldehyde (25.00g, 201.43mmol,21.19ml, 1.00eq) is slowly dropwise added into the reaction system, the system slowly rises to room temperature within 30 minutes, a saturated ammonium chloride (100 mL) solution is added into the system, water (200 mL) is added, ethyl acetate (150ml × 3) is used for extraction, organic phases are combined, the organic phases are washed by saturated saline water (150 mL), dried by sodium sulfate, filtered, concentrated, and separated and purified by a column (silica gel column, petroleum ether: ethyl acetate = 5: 1) to obtain the compound 16-2. ¹ H NMR(400MHz，CDCl ₃ ) δ 8.30 (d, J =5.2hz, 1h), 7.43-7.37 (m, 1H), 7.35-7.29 (m, 2H), 7.20 (dd, J =0.8,5.2hz, 1h), 7.11-7.03 (m, 2H), 5.78 (d, J =2.6hz, 1h), 2.67 (d, J =3.4hz, 1h), MS ESI calculated value C ₁₂ H ₉ ClFNO[M+H] ⁺ 237.66, found 238.2.

And 2, step: to compound 16-2 (23.00g, 96.78mmol, 1.00eq) in dichloromethane (230.00 mL) was added dess-martin oxidant (45.15g, 106.46mmol,32.96mL, 1.10eq) at 0 ℃ and the system was warmed to 25 ℃ and stirred for two hours. The mixed system was filtered, and the filtered filtrate was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to obtain compound 16-3.MS ESI calculated value C ₁₂ H ₇ ClFNO[M+H] ⁺ 235.64, found 236.2.

And step 3: to a solution of compound 16-3 (3.00g, 12.73mmol, 1.00eq) in toluene (100.00 mL) was addedP-toluenesulfonic acid (7.27g, 38.19mmol, 3.00eq) and ethylene glycol (1.58g, 25.46mmol,1.42mL, 2.00eq) were added, and the reaction was heated to 110 ℃ and stirred for 5 hours. The mixture was concentrated and column purified (petroleum ether: ethyl acetate = 10: 1 to 5: 1) to give compound 16-4.MS ESI calculated value C ₁₄ H ₁₁ ClFNO ₂ [M+H] ⁺ 279.69, found 280.0.

And 4, step 4: to a mixture of compounds 16-4 (100.00mg, 336.09. Mu. Mol,1.00 eq) in 1, 4-dioxane (5.00 mL) were added tris (dibenzylideneacetone) dipalladium (61.55mg, 67.22. Mu. Mol,0.20 eq), potassium hydroxide (40.12mg, 715.08. Mu. Mol,2.00 eq) and 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (28.54mg, 67.22. Mu. Mol,0.20 eq), and the system was heated to 90 ℃ and stirred for 3 hours. The mixture was concentrated, water (10 mL) was added, extraction was performed with ethyl acetate (10 mL × 3), and the organic phases were combined, washed with saturated brine (10 mL), dried over sodium sulfate, filtered, concentrated, and purified to give compound 16-5.MS ESI calculated value C ₁₄ H ₁₂ FNO ₃ [M+H] ⁺ 261.25, found 262.2.

And 5: to a mixture of compounds 16-5 (80.00mg, 281.72. Mu. Mol,1.00 eq) in N, N-dimethylformamide (10.00 mL) was added sodium hydrogen (22.54mg, 563.44. Mu. Mol,60% purity, 2.00 eq) under nitrogen at 0 ℃ and after stirring the system for ten minutes, methyl 4-bromomethylbenzoate (129.07mg, 563.44. Mu. Mol,2.00 eq) was added and stirring was continued for thirty minutes. The mixture was quenched with water (5 mL), extracted with ethyl acetate (5 mL × 3), and the combined organic phases were washed with water (10 mL), saturated brine (10 mL), dried over sodium sulfate, filtered, concentrated and purified to give compound 16-6.MS ESI calculated value C ₂₃ H ₂₀ FNO ₅ [M+H] ⁺ 409.41, found 410.3.

Step 6: to a solution of compound 16-6 (70.00mg, 141.91. Mu. Mol,1.00 eq) in methanol (5.00 mL) were added an aqueous hydroxylamine solution (1.00mL, 50% purity) and sodium hydroxide (17.03mg, 425.73. Mu. Mol,3.00 eq) and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated and purified by preparative HPLC (0.225% by weight FA) to give compounds 16-7. ¹ H NMR(400MHz，DMSO-d ₆ )δ＝7.77(d，J＝7.2Hz，1H)，7.68 (d, J =8.2hz, 2h), 7.53-7.45 (m, 2H), 7.33 (d, J =8.2hz, 2h), 7.19 (t, J =8.8hz, 2h), 6.48 (d, J =1.8hz, 1h), 6.20 (dd, J =1.8,7.2hz, 1h), 5.08 (s, 2H), 3.99 (m, 4H). MS ESI calcd C ₂₂ H ₁₉ FN ₂ O ₅ [M+H] ⁺ 410.40, found 411.1.

Example 17

Step 1: in N ₂ Under protection, a solution of methyltriphenylphosphonium iodide (2.83g, 7.01mmol,1.10 eq) and t-BuOK (1M, 7.01mL,1.10 eq) in THF (20.00 mL) was heated under reflux for 1 hour. The suspension was then cooled to 0 ℃ and compound 17-1 (1.50g, 6.37mmol, 1.00eq) was added and the reaction stirred at 20 ℃ for 12 h. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (50 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was isolated and purified by silica gel column chromatography (PE/EtOAc = 20/1) to give compound 17-2.MS ESI calculated value C ₁₃ H ₉ ClFN[M+H]+234, found 234.

And 2, step: mixing the compound 17-2 (800.00mg, 3.42mmol, 1.00eq) and OsO ₄ (869.47mg, 3.42mmol, 177.44. Mu.L, 1.00 eq) in THF (10.00 mL) and H ₂ The O (2.00 mL) solution was stirred at 20 ℃ for 4 hours. Then the reaction solution was treated with 2M Na ₂ SO ₃ The aqueous solution (80 mL) was washed with water and extracted with EtOAc (30mL x3), and the combined extract phases were washed with brine (50 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain compound 17-3.MS ESI calculated value C ₁₃ H ₁₁ ClFNO ₂ [M+H]+268, found value 268.

And 3, step 3: the compound 17-3 (800.00mg, 2.99mmol, 1.00eq), 1,2-dibromoethane (561.45mg, 2.99mmol, 225.48. Mu.L, 1.00 eq) and t-BuOK (503.03mg, 4.49mmol, 1.50eq) in DMSO (10.00 mL) were heated to 70 ℃ and stirred for 6 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc/MeOH (10/1, 50mL x3), and the combined extracts were washed with brine (80 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting solid was isolated and purified by preparative TLC (PE/EtOAc = 2/1) to afford compound 17-4.MS ESI calculated value C ₁₅ H ₁₃ ClFNO ₂ [M+H]+294, found 294.

And 4, step 4: in N ₂ Under the protection, compound 17-4 (180.00mg, 612.83. Mu. Mol,1.00 eq), t-BuXPhOS (26.02mg, 61.28. Mu. Mol,0.10 eq), pd ₂ (dba) ₃ (56.12mg, 61.28. Mu. Mol,0.10 eq) and KOH (68.77mg, 1.23mmol, 2.00eq) in dioxane (10.00 mL) and H ₂ The O (2.00 mL) solution was heated to 100 ℃ and stirred for 1 hour. The reaction solution was poured into H ₂ O (80 mL) and adjusted pH =6 with 2M HCl, etoac (30mL × 3) extraction, combined extract phase washed with brine (50 mL) and then anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was isolated and purified by preparative TLC (PE/EtOAc = 3/1) to afford compound 17-5.MS ESI calculated value C ₁₅ H ₁₄ FNO ₃ [M+H]+276, found 276.

And 5: at N ₂ To a solution of compound 17-5 (90.00mg, 326.95. Mu. Mol,1.00 eq) in DMF (5.00 mL) under protection, naH (15.69mg, 392.34. Mu. Mol,60% purity, 1.20 eq) was added at 0 ℃ and stirring continued at this temperature for 20 minutes. Methyl 4- (bromomethyl) benzoate (74.89mg, 326.95. Mu. Mol,1.00 eq) was then added to the suspension, and the resulting reaction mixture was heated to 15 ℃ and stirred for 1 hour. The reaction solution was poured into H ₂ O (50 mL) and extracted with EtOAc (20mL. Times.3), and the combined extract was washed with brine (50 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting solid was isolated and purified by preparative TLC (PE/EtOAc = 1/2) to afford compound 17-6.MS ESI calculated value C ₂₄ H ₂₂ FNO ₅ [M+H]+424, found value 424.

Step 6: to compound 17-6 (100.00mg, 236.17. Mu. Mol,1.00 eq) in DCM (1) at 0 ℃.00 mL) and MeOH (2.00 mL) in sequence ₂ OH.H ₂ O (1.00mL, 50% aqueous solution) and aqueous NaOH (2M, 1.00mL). After completion of the dropwise addition, the reaction solution was stirred at 0 ℃ for 6 hours. The solvent was removed by concentration under reduced pressure. The remaining solution was cooled to 0 ℃ and adjusted pH =8 with 2M HCl, filtered and diluted with H ₂ And repeatedly rinsing O (5mL x3), and finally drying in vacuum to obtain the compound 17-7. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.17 (brs, 1H), 9.04 (brs, 1H), 7.74 (d, J =7.28hz, 1h), 7.69 (d, J =8.28hz, 2h), 7.46 (dd, J =9.03,5.52hz, 2h), 7.33 (d, J =8.28hz, 2h), 7.19 (t, J =8.91hz, 2h), 6.57 (d, J =1.76hz, 1h), 6.13 (dd, J =7.15,1.88hz, 1h), 5.07 (s, 2H), 4.11-4.24 (m, 1H), 3.99-4.08 (m, 1H), 3.54-3.71 (m, 4H) · MS ESI calculated value C ₂₃ H ₂₁ FN ₂ O ₅ [M+H]+425, measured value 425.

Example 18

Step 1: to a solution of 1, 4-butyrolactone (3.45g, 40.11mmol,3.05mL, 1.10eq) in toluene (30.00 mL) at 0 ℃ was added sodium hydride (1.75g, 43.75mmol, purity 60%,1.20 eq) and a solution of compound 18-1 (5.00g, 36.46mmol,4.39mL, 1.00eq) in toluene (10.00 mL) at 0 ℃ and the reaction was stirred at 20 ℃ for 12 hours. Saturated aqueous ammonium chloride solution and water were added to the reaction solution, followed by extraction with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give compound 18-2 which was used directly in the next reaction.

And 2, step: a mixture containing 18-2 (3.22g, 16.84mmol, 1.00eq) and 36% hydrochloric acid (10.20g, 100.71mmol,10.00mL, 5.98eq) was heated to 80 ℃ and reacted for 3 hours with stirring. The reaction solution was poured into a saturated aqueous solution of potassium carbonate at 0 ℃, water was added, extraction was performed with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified with silica gel column to obtain compound 18-3 which was used directly in the next reaction.

And step 3: at-68 ℃, an n-butyllithium-n-hexane solution (2.5M, 1.34mL, 1.10eq) was added dropwise to a toluene (5.00 mL) solution of 2-chloro-4-iodo-pyridine (730.20mg, 3.05mmol, 1.00eq), and then a toluene (1.00 mL) solution of the compound 18-3 (560.00mg, 3.05mmol, 1.00eq) was added, and the reaction was gradually warmed to 20 ℃ and stirred for 2 hours. Adding saturated ammonium chloride aqueous solution into the reaction solution, water, extracting with ethyl acetate, washing the organic phase with saturated saline solution, drying with anhydrous sodium sulfate, filtering, concentrating, and purifying with silica gel column to obtain compound 18-4.MS ESI calculated value C ₁₄ H ₁₃ ClN ₂ O[M+H]+261.0, found 261.0.

And 4, step 4: to a solution of compound 18-4 (750.00mg, 2.88mmol, 1.00eq) in dioxane (5.00 mL) and water (1.00 mL) were added potassium hydroxide (322.82mg, 5.75mmol, 2.00eq), tris (dibenzylideneacetone) dipalladium (263.42mg, 287.66. Mu. Mol,0.10 eq), 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (122.15mg, 287.66. Mu. Mol,0.10 eq), and stirred at 100 ℃ for 4 hours. To the reaction solution were added saturated ammonium chloride and water, followed by extraction with ethyl acetate, washing with saturated brine, drying over anhydrous sodium sulfate, filtration and concentration. Concentrating, purifying with silica gel column to obtain compound 18-5.MS ESI calculated value C ₁₄ H ₁₄ N ₂ O ₂ [M+H]+243, measured value 243.

And 5: to a solution of compound 18-5 (163.00mg, 672.80. Mu. Mol,1.00 eq) in N, N-dimethylformamide (2.00 mL) was added sodium hydride (32.29mg, 807.36. Mu. Mol,60% purity, 1.20 eq) at 0 ℃ and then a solution of methyl 4-bromomethylbenzoate (154.12mg, 672.80. Mu. Mol,1.00 eq) in N, N-dimethylformamide (1.00 mL) was added to the reaction system, followed by stirring at 20 ℃ for 2 hours. Saturated aqueous ammonium chloride solution and water were added to the reaction mixture, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to give compound 18-6.MS ESI calculated value C ₂₃ H ₂₂ N ₂ O ₄ [M+H]+391.0, found 391.0.

And 6: to a solution of compound 18-6 (220.00mg, 563.48. Mu. Mol,1.00 eq) in methanol (2.00 mL) was added sodium hydroxide (22.54mg, 563.48. Mu. Mol,1.00 eq) and an aqueous solution of hydroxylamine (1.00mL, 50% aqueous solution), and the mixture was stirred at 20 ℃ for 6 hours. The reaction was concentrated and purified by reverse phase preparative HPLC to give compound 18-7. ¹ H NMR (400mhz, metanol-d 4) δ =8.53 (dd, J =0.8,4.8hz, 1h), 7.82-7.74 (m, 1H), 7.70 (d, J =8.2hz, 2h), 7.67 (d, J =7.9hz, 1h), 7.60 (d, J =7.1hz, 1h), 7.36 (d, J =8.3hz, 2h), 7.27 (ddd, J =1.1,4.9,7.5hz, 1h), 6.79 (d, J =1.8hz, 1h), 6.60 (dd, J =2.0,7.1hz, 1h), 5.18 (s, 2H), 4.17-3.96 (m, 2H), 3.04-2.91 (m, 1H), 2.38 (td, J =7.6, 12.6hz, 1h), 2.06-1.80 (m, 2H), MS ESI calculated value C ₂₂ H ₂₁ N ₃ O ₄ [M+H]+392, found 392.

Example 19

Step 1: to a solution of 1, 4-butyrolactone (1.00g, 11.62mmol, 884.96. Mu.L, 1.00 eq) in tetrahydrofuran (5.00 mL) at 0 ℃ was added sodium hydride (1.06g, 26.38mmol, 60% purity, 2.27 eq), and the mixture was stirred at 0 ℃ for 30 minutes, followed by addition of a solution of compound 19-1 (2.39g, 17.43mmol,2.06mL, 1.50eq) in tetrahydrofuran (2.00 mL) at 0 ℃ and stirring at 20 ℃ for 2 hours. To the reaction mixture were added saturated aqueous ammonium chloride solution, water and ethyl acetate. The water phase is concentrated and dried, acetone is added, stirring and filtering are carried out, and the mother liquor is concentrated and dried to obtain the compound 19-2 which is directly used for the next reaction.

Step 2: a mixture containing 19-2 (1.50g, 7.85mmol, 1.00eq) and 36% hydrochloric acid (15.30g, 151.11mmol,15.00mL, 19.26eq) was heated to 80 ℃ and stirred for 2 hours. The reaction solution was poured into ice water at 0 ℃ and extracted with ethyl acetate, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified with silica gel column to give compound 19-3 which was used directly in the next reaction.

And 3, step 3: at-68 deg.C to 2-chloro-4A solution of iodine-pyridine (782.97mg, 3.27mmol, 1.00eq) in toluene (5.00 mL) was added dropwise to a solution of n-butyllithium in n-hexane (2.5M, 1.44mL, 1.10eq) followed by a solution of compound 19-3 (600.00mg, 3.27mmol, 1.00eq) in toluene (1.00 mL), the reaction was gradually warmed to 20 ℃ and stirred for 12 hours. Adding saturated ammonium chloride aqueous solution into the reaction solution, water, extracting with ethyl acetate, washing the organic phase with saturated saline solution, drying with anhydrous sodium sulfate, filtering, concentrating, and purifying with silica gel column to obtain compound 19-4.MS ESI calculated value C ₁₄ H ₁₃ ClN ₂ O[M+H]+261.0, found 261.0.

And 4, step 4: to a solution of compound 19-4 (513.00mg, 1.97mmol, 1.00eq) in dioxane (5.00 mL) and water (1.00 mL) were added potassium hydroxide (221.07mg, 3.94mmol, 2.00eq), tris (dibenzylideneacetone) dipalladium (180.40mg, 197.00. Mu. Mol,0.10 eq) and 2-di-tert-butylphosphine-2 ',4',6' -triisopropylbiphenyl (83.65mg, 197.00. Mu. Mol,0.10 eq), and stirred at 100 ℃ for 2 hours. Water was added to the reaction mixture, which was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. Concentrating, purifying with silica gel column to obtain compound 19-5.MS ESI calculated value C ₁₄ H ₁₄ N ₂ O ₂ [M+H]+243, measured value 243.

And 5: to a solution of 19 to 5 (240.00mg, 990.63. Mu. Mol,1.00 eq) in N, N-dimethylformamide (5.00 mL) at 0 ℃ was added sodium hydride (47.55mg, 1.19mmol, purity 60%,1.20 eq), and then to the reaction system was added a solution of methyl 4-bromomethylbenzoate (226.92mg, 990.63. Mu. Mol,1.00 eq) in N, N-dimethylformamide (1.00 mL), and the mixture was stirred at 20 ℃ for 2 hours. Adding saturated ammonium chloride aqueous solution and water into the reaction solution, extracting with ethyl acetate, washing with saturated saline solution, drying with anhydrous sodium sulfate, concentrating, and purifying with chromatography plate to obtain compound 19-6.MS ESI calculated value C ₂₃ H ₂₂ N ₂ O ₄ [M+H]+391.1, found 391.1

And 6: to a solution of compound 19-6 (66.00mg, 169.04. Mu. Mol,1.00 eq) in methanol (5.00 mL) were added sodium hydroxide (13.52mg, 338.08. Mu. Mol,2.00 eq) and aqueous hydroxylamine solution (1.00mL, 50% purity), and the mixture was stirred at 20 ℃ for 12 hours. After the reaction solution is concentrated,purifying by reverse phase preparative column to obtain the compound 19-7. ¹ H NMR (400mhz, meod) δ 8.51 (d, J =6.1hz, 2h), 8.45 (brs, 1H), 8.47-8.41 (m, 1H), 7.71 (d, J =8.3hz, 2h), 7.65 (d, J =7.2hz, 1h), 7.62-7.56 (m, 2H), 7.37 (d, J =8.3hz, 2h), 6.76 (d, J =1.9hz, 1h), 6.49 (dd, J =1.9,7.2hz, 1h), 5.20 (s, 2H), 4.13-3.99 (m, 2H), 2.58 (t, J =7.2hz, 2h), 2.09-1.88 (m, 2H), MS ESI C calculated value ₂₂ H ₂₁ N ₃ O ₄ [M+H]+392, found 392.

Example 20

Step 1: at N ₂ Protection n-BuLi (2.5M, 23.33mL, 1.10eq) was added dropwise to a solution of 1-bromo-4-iodobenzene (15.00g, 53.02mmol, 1.00eq) in THF (150.00 mL) at-70 ℃ and, after completion of the addition, stirring was continued at that temperature for 30 minutes. Then, compound 20-1 (10.64g, 53.02mmol,8.72mL, 1.00eq) was added dropwise to the above yellow suspension, and after completion of the dropwise addition, the reaction system was gradually warmed to 20 ℃ and stirred for 12 hours. After completion of the reaction, water (300 mL) was added and quenched and extracted with EtOAc (100mL. Times.3), and the combined extracts were washed with brine (100 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain compound 20-2. ¹ H NMR(400MHz，CDCl ₃ )δ7.25-7.50(m，6H)，6.90-7.00(m，2H)，4.03(t，J＝6.8Hz，2H)，2.40-2.55(m，2H)，1.85-2.00(m，2H).

Step 2: in N ₂ Protection n-BuLi (2.5M, 1.24mL, 1.00eq) was added dropwise at-70 ℃ to a solution of compound 20-2 (1.00g, 3.11mmol, 1.00eq) in THF (15.00 mL) and, after completion of the addition, stirring was continued at this temperature for 30 minutes. Dry ice (about 5 g) was then added to the reaction system and gradually warmed to 20 ℃ with stirring for 2 hours. After completion of the reaction, the reaction solution was poured into H ₂ O (30 mL), pH =9 with 2M KOH, and extracted with EtOAc (10mL × 2). The aqueous phase was then adjusted to pH =5 with 2M HCl and extracted with EtOAc (20mL × 3). The extract was washed with brine (30 mL) and then with anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain compound 20-3. ¹ H NMR(400MHz，DMSO-d ₆ )δ7.87(d，J＝8.0Hz，2H)，7.56(d，J＝7.6Hz，2H)，7.40-7.50(m，2H)，7.12(t，J＝8.8Hz，2H)，3.90-4.00(m，2H)，2.51-2.60(m，2H)，1.75-1.90(m，2H).

And step 3: a solution of compound 20-3 (100.00mg, 349.28. Mu. Mol,1.00 eq), HATU (159.37mg, 419.14. Mu. Mol,1.20 eq) and DIEA (67.71mg, 523.92. Mu. Mol, 91.50. Mu.L, 1.50 eq) in MeCN (10.00 mL) was stirred at 20 ℃ for 30 minutes, then methyl 4- (aminomethyl) benzoate (70.43mg, 349.28. Mu. Mol,1.00eq, HCl salt) was added to the above solution and stirring was continued at 20 ℃ for 12 hours. After the reaction is completed, the reaction solution is concentrated, and the residue is treated with H ₂ Pulping with O (3 mL), filtering, and vacuum drying to obtain compound 20-4, which is directly used in the next step. MS ESI calculated value C ₂₆ H ₂₄ FNO ₄ [M+H] ⁺ 434, found 434.

And 4, step 4: to a solution of compound 20-4 (150.00mg, 346.04. Mu. Mol,1.00 eq) in DCM (4.00 mL) and MeOH (2.00 mL) was added NH dropwise in that order at 20 deg.C ₂ OH.H ₂ O (3.00mL, 50% aqueous solution) and aqueous NaOH (2M, 1.50mL). After completion of the dropwise addition, the reaction solution was heated under reflux for 6 hours. Then directly separated and purified by preparative HPLC (0.1% TFA) to give 20-5. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.15 (brs, 1H), 8.95-9.05 (m, 2H), 7.80 (d, J =8.0hz, 2h), 7.67 (d, J =8.0hz, 2h), 7.52 (d, J =8.4hz, 1h), 7.40-7.50 (m, 2H), 7.33 (d, J =8.4hz, 2h), 7.02-7.10 (m, 2H), 4.67 (d, J =5.6hz, 2h), 3.92 (t, J =6.8hz, 2h), 2.51-2.60 (m, 2H), 1.85-1.95 (m, 2H). MS calculated value C ESI ₂₅ H ₂₃ FN ₂ O ₄ [M+H]+435, found 435.

Example 21

Step 1: compound 21-1 (1g, 2.31mmol) was separated by preparative SFC (chiral separation column model: OJ (250mm. About.30mm, 5 um)) Eluent: a: supercritical fluid CO 2B: 0.1% of NH ₃ H ₂ O MeOH, increasing the B phase from 5% to 40% in 5.5 minutes and keeping the B phase at 40% for 3 minutes, finally 5% B phase was kept for 1.5 minutes at a flow rate of 60 mL/min), resulting in two isomers 21-2 (P1) and 22-1 (P2) with retention times of 6.474min and 7.256min, respectively.

Step 2: to a solution of compound 21-2 (P1) (450.00mg, 1.04mmol, 1.00eq) in DCM (4.00 mL) and MeOH (4.00 mL) was added NH, one after the other, dropwise at 0 deg.C ₂ OH.H ₂ O (4.00mL, 50% aqueous solution) and NaOH (2M, 4.00mL) were added dropwise and stirring was continued at this temperature for 2 hours. Concentrating under reduced pressure to remove most of the solvent, cooling the remaining solvent to 0 deg.C and adjusting pH =7-8 with 8M HCl, filtering the precipitated solid, and filtering the filter cake with H ₂ O (5mL. Multidot.2) and then dried under reduced pressure to obtain the compound 21-3. ¹ H NMR (400mhz, meod) δ 7.79 (d, J =8.4hz, 2h), 7.71 (d, J =8.4hz, 2h), 7.54 (d, J =8.4hz, 2h), 7.40-7.05 (m, 4H), 6.95-7.05 (m, 2H), 4.60 (d, J =4.0hz, 2h), 4.02 (t, J =7.2hz, 2h), 2.50-2.60 (m, 2H), 1.85-2.00 (m, 2H). MS ESI calcd C ₂₅ H ₂₃ FN ₂ O ₄ [M+H] ⁺ 435, found 435.

Example 22

Step 1: to a solution of compound 22-1 (P2) (450.00mg, 1.04mmol, 1.00eq) in DCM (4.00 mL) and MeOH (4.00 mL) at 0 deg.C was added dropwise NH in that order ₂ OH.H ₂ O (4.00mL, 50% aqueous solution) and NaOH (2M, 4.00mL) were added dropwise and stirring was continued at this temperature for 2 hours. Concentration under reduced pressure removed most of the solvent, cooling the remaining solvent to 0 ℃ and adjusting pH =7-8 with 8M HCl, and work-up afforded compound 22-2 (P2). ¹ H NMR (400mhz, meod) δ 7.79 (d, J =8.4hz, 2h), 7.71 (d, J =8.4hz, 2h), 7.54 (d, J =8.4hz, 2h), 7.40-7.05 (m, 4H), 6.95-7.05 (m, 2H), 4.60 (s, 2H), 4.02 (t, J =7.2hz, 2h), 2.50-2.60 (m, 2H), 1.85-2.00 (m, 2H). MS ESI calculated value C ₂₅ H ₂₃ FN ₂ O ₄ [M+H] ⁺ 435, found 435.

Example 23

Step 1: a mixture of compound 23-1 (1.00g, 3.11mmol, 1.00eq), tert-butoxycarbonylamino (546.50mg, 4.67mmol, 1.50eq), xantphos (179.95mg, 311.00. Mu. Mol,0.10 eq), pd ₂ (dba) ₃ (142.39mg, 155.50. Mu. Mol,0.05 eq) and Cs ₂ CO ₃ (1.52g, 4.67mmol, 1.50eq) in DMF (15.00 mL) in N ₂ Heating to 100 ℃ under protection and stirring for 12 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (80 mL) and then over anhydrous Na ₂ Drying SO4, filtering, and concentrating under reduced pressure. The resulting product was subjected to silica gel column chromatography (PE/EtOAc = 9/1) to afford compound 23-2.MS ESI calculated value C ₂₁ H ₂₄ FNO ₃ [M+H]+358, found 358.

Step 2: to a solution of compound 23-2 (250.00mg, 699.46. Mu. Mol,1.00 eq) in DCM (5.00 mL) was added TFA (2.00 mL) at 25 ℃ and stirred for 30 min. Then carrying out reduced pressure concentration to obtain the compound 23-3.MS ESI calculated value C ₁₆ H ₁₆ FNO[M+H] ⁺ 258, found 258.

And step 3: compounds 23-3 (250.00mg, 673.26. Mu. Mol,1.00eq, TFA salt), methyl 4- (bromomethyl) benzoate (154.22mg, 673.26. Mu. Mol,1.00 eq) and K ₂ CO ₃ A solution of (139.58mg, 1.01mmol, 1.50eq) in DMF (5.00 mL) was stirred at 25 ℃ for 6 h. The reaction solution was then poured into H ₂ O (50 mL) and extracted with EtOAc (30mL. Times.2), and the combined extracts were washed with brine (30 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain compound 23-4 which can be directly used in the next reaction. MS ESI calculated value C ₂₅ H ₂₄ FNO ₃ [M+H] ⁺ 406, measured value 406.

And 4, step 4: to compound 23-4 (250.00mg, 616.58. Mu. Mol,1.00 eq) in DCM (4) at 25 deg.C00 mL) and MeOH (2.00 mL) in sequence ₂ OH.H ₂ O (3.00mL, 50% aqueous solution) and NaOH (2M, 1.50mL) in water, and the reaction was warmed to 50 ℃ and stirred for 2 hours. Concentrating under reduced pressure to remove most of the solvent, and subjecting the remaining solution to preparative HPLC (0.1% ₄ OH) is separated and purified to obtain the compound 23-5. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.04 (brs, 1H), 9.03 (brs, 1H), 7.67 (d, J =8.0hz, 2h), 7.35-7.45 (m, 4H), 6.95-7.05 (m, 4H), 6.45 (d, J =8.4hz, 2h), 6.28 (t, J =5.6hz, 1h), 4.25 (d, J =6.0hz, 2h), 3.70-3.90 (m, 2H), 2.20-2.45 (m, 4H), 1.65-1.90 (m, 2H). MS ESI calcd C ₂₅ H ₂₄ FNO ₃ [M+H]+406, found 406.

Example 24

Step 1: compounds No. 24-1 (300.00mg, 934.06. Mu. Mol,1.00 eq), methyl 4-aminobenzoate (141.19mg, 934.06. Mu. Mol,1.00 eq), xantphos (108.09mg, 186.81. Mu. Mol,0.20 eq), cs ₂ CO ₃ (456.50mg, 1.40mmol, 1.50eq) and Pd ₂ (dba) ₃ (85.53mg, 93.41. Mu. Mol,0.10 eq) in DMF (15.00 mL) in N ₂ Heated to 100 ℃ under protection and stirred for 8 hours. The reaction solution was poured into H ₂ O (150 mL) and extracted with EtOAc (80mL. Times.2), and the combined extracts were washed with brine (50 mL) and then over anhydrous Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure. The obtained product is separated and purified by silica gel column chromatography (PE/EtOAc = 5/1) to obtain the compound 24-2.MS ESI calculated value C ₂₄ H ₂₂ FNO ₃ [M+H]+392, found 392.

Step 2: to a solution of compound 24-2 (250.00mg, 638.67. Mu. Mol,1.00 eq) in DCM (2.00 mL) and MeOH (4.00 mL) was added dropwise, in that order, NH2OH.H2O (2.00mL, 50% aqueous solution) and NaOH (2M, 2.00mL) aqueous solution at 25 ℃. After completion of the dropwise addition, the reaction solution was further stirred at 20 ℃ for 12 hours. Concentrating under reduced pressure to remove most of the solvent, then dissolving in DMSO (5 mL), and subjecting the resulting solution to preparative HPLC (0.1% NH) ₄ OH) is separated and purified to obtain the compound 24-3.

¹ H NMR (400MHz, DMSO-d 6) delta 10.88 (brs, 1H), 8.85 (brs, 1H), 8.50 (brs, 1H), 6.90-7.80 (m, 12H), 3.80-4.00 (m, 2H), 2.30-2.45 (m, 2H), 1.80-1.90 (m, 2H). MS ESI calcd for C ₂₃ H ₂₁ FN ₂ O ₃ [M+H]+393, found 393.

Example 25

Step 1: a solution of compound 25-1 (800.00mg, 2.79mmol, 1.00eq), HATU (1.59g, 4.19mmol, 1.50eq) and DIEA (722.26mg, 5.59mmol, 976.03. Mu.L, 2.00 eq) in MeCN (5.00 mL) was stirred at 20 ℃ for 20 minutes, then methyl 4-aminobenzoate (422.38mg, 2.79mmol, 1.00eq) was added to the solution and stirring continued at this temperature for 12 hours. After removal of the solvent by concentration under reduced pressure, the remaining solid was dissolved in EtOAc (80 mL), washed successively with 2M aqueous NaOH (30mL. Times.2), brine (50 mL), and then washed with anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. The resulting product was subjected to silica gel column chromatography (PE/EtOAc = 10/1) to afford compound 25-2.MS ESI calculated value C ₂₅ H ₂₂ FNO ₄ [M+H]+420, found 420.

Step 2: to a solution of compound 25-2 (1.10 g,2.62mmol, 1.00eq) in DCM (10.00 mL) and MeOH (20.00 mL) was added NH dropwise in that order at 20 deg.C ₂ OH.H ₂ O (11.00mL, 50% aqueous solution) and NaOH (2M, 10.00mL) in water, and stirring was continued at this temperature for 12 hours after the addition was complete. Most of the solvent was removed by concentration under reduced pressure, then dissolved in DMSO (5 mL), and the resulting solution was purified by preparative HPLC (0.1% HCl) to obtain Compound 25-3.

¹ H NMR(400MHz，DMSO-d ₆ )δ11.14(brs，1H)，10.39(brs，1H)，7.86(d，J＝8.4Hz，2H)，7.82(d，J＝8.4Hz，2H)，7.40(d，J＝8.8Hz，2H)，7.61(d，J＝8.8Hz，2H)，7.45-7.55(m，2H)，7.13(t，J＝8.8Hz，2H)，3.96(t，J＝7.6Hz，2H)，2.58(t，J＝6.8Hz，2H) MS ESI calculated value C of 1.80-1.95 (m, 2H) ₂₄ H ₂₁ FN ₂ O ₄ [M+H]+421, measured value 421.

Example 26

Step 1: in N ₂ Protection n-BuLi (2.5M, 12.47mL, 1.20eq) was added dropwise to a solution of compound 26-1 (5.00g, 25.98mmol, 1.00eq) in toluene (50.00 mL) at-70 ℃ and, after completion of the addition, stirring was continued at that temperature for 30 minutes. 4-chloro-1- (4-fluorophenyl) butyl-1-one (5.21g, 25.98mmol,4.27mL, 1.00eq) was then added to the yellow suspension. And the reaction was warmed to 15 ℃ and stirred for 12 hours. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (80 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering and concentrating under reduced pressure. The obtained product is separated and purified by silica gel column chromatography (PE/EtOAc = 10/1) to obtain the compound 26-2.MS ESI calculated value C ₁₅ H ₁₃ ClFNO[M+H]+278, actual measurement value 278.

Step 2: mixing the compound 26-2 (1.00g, 3.60mmol, 1.00eq), DPPP (148.51mg, 360.00. Mu. Mol,0.10 eq), pd ₂ (dba) ₃ A mixture of (164.86mg, 180.00. Mu. Mol,0.05 eq) and TEA (546.54mg, 5.40mmol, 748.68. Mu.L, 1.50 eq) in DMSO (10.00 mL) and MeOH (3.00 mL) was heated to 80 ℃ and stirred under an atmosphere of CO (50 psi) for 12 h. The reaction solution was poured into H ₂ O (100 mL) and extracted with EtOAc (50mL. Times.3), and the combined extracts were washed with brine (80 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure to obtain compound 26-3.MS ESI calculated value C ₁₇ H ₁₆ FNO ₃ [M+H]+302, found 302.

And 3, step 3: compounds 26-3 (1.00g, 3.32mmol, 1.00eq) and NaOH (265.60mg, 6.64mmol, 2.00eq) in MeOH (10.00 mL) and H ₂ The O (10.00 mL) mixture solution was heated to reflux and stirred for 2 hours. MeOH was then removed by concentration under reduced pressure, the resulting aqueous phase was extracted with EtOAc (10mL. Times.2), and then water was addedAdjusting pH of the obtained phase with concentrated hydrochloric acid =6, and concentrating under reduced pressure to obtain compound 26-4.MS ESI calculated value C ₁₆ H ₁₄ FNO ₃ [M+H]+288, found 288.

And 4, step 4: a solution of 26-4 (900.00mg, 3.13mmol, 1.00eq), HATU (1.79g, 4.70mmol, 1.50eq) and DIEA (1.21g, 9.40mmol, 3.00eq) in MeCN (15.00 mL) was stirred at 20 ℃ for 30 minutes. Methyl 4- (aminomethyl) benzoate (631.71mg, 3.13mmol,1.00eq, HCl salt) was then added to the above solution and stirring was continued at 20 ℃ for 3 hours. Concentrating under reduced pressure to dry. Using water H ₂ O (80 mL) was diluted and then extracted with EtOAc (30mL x3), and the combined extract was washed with brine (30 mL) and over anhydrous Na ₂ SO ₄ Drying, filtering, and concentrating under reduced pressure. And separating and purifying by silica gel column chromatography (PE/EtOAc =5/1 to 3/1) to obtain 26-5.MS ESI calculated value C ₂₅ H ₂₃ FN ₂ O ₄ [M+H]+435, found 435.

And 5: to a solution of 26-5 (200.00mg, 460.34. Mu. Mol,1.00 eq) in DCM (2.00 mL) and MeOH (4.00 mL) at 0 ℃ is added NH in succession ₂ OH.H ₂ O (47.04mg, 460.34. Mu. Mol,2.00mL,50% aqueous solution) and NaOH (2M, 230.17. Mu.L) aqueous solution, and stirring was continued at this temperature for 3 hours after the addition was complete. Concentrating under reduced pressure to remove most of the solvent, dissolving in DMSO (5 mL), and separating and purifying the solution by preparative HPLC (0.05% HCl) to obtain 26-6. ¹ H NMR(400MHz，DMSO-d ₆ ) δ 11.15 (brs, 1H), 10.22 (brs, 1H), 9.35 (t, J =6.4hz, 1h), 8.72 (d, J =1.5hz, 1h), 8.01-8.07 (m, 1H), 7.94-8.00 (m, 1H), 7.68 (d, J =8.3hz, 2h), 7.49-7.56 (m, 2H), 7.34 (d, J =8.3hz, 2h), 7.15 (t, J =8.9hz, 2h), 4.51 (d, J =6.3hz, 2h), 3.98 (t, J =7.2hz, 2h), 2.62 (t, J =7.2hz, 2qh), 1.89 (d, J =7.2,5.1hz, 2h), MS calculated value C ₂₄ H ₂₂ FN ₃ O ₄ [M+H]+436, found 436.

Experimental example 1: in vitro evaluation

By passingMultifunctional microplate reader for detecting fluorescence in 384 microporous plateThe change of signal intensity is used for measuring the deacetylation degree of an acetylated substrate, and the inhibition effect of the compound on Histone Deacetylase (HDAC) is evaluated by taking the half inhibition concentration IC50 value of the compound inhibitor as an index.

1. Experimental procedures and methods:

1.1 dilution of Compound Loading:

compounds were diluted to 2mM with DMSO into 384 compound plates, 3-fold diluted using Bravo full-automatic liquid workstation, 10 gradients, double wells; the Echo ultrasonic liquid workstation equipment (Echo liquid handler) then transferred 0.15. Mu.L of compound from the 384 compound plate to the 384 lab plate.

1.2 Assay buffer configuration: the preparation contained 20mM hepes,137mM NaCl,2.7mM KCl, and 1mM MgCl at the final concentration ₂ 0.05% BSA, 1 × assay buffer of 0.5mM TCEP.

1.3 1.5 Xpreparation of enzyme mixture:

the fast dissolving histone deacetylase was added to a 1 × assay buffer and mixed well. Add 1.5 Xthe enzyme mixture to 20. Mu.L to 384 well assay plates, mix by centrifugation, place in a 23 ℃ incubator, and incubate the enzyme with the compound for 20 minutes.

1.43 × preparation of substrate mixture:

prepare 3 × substrate solution with 1 × assay buffer, mix well. The mixture was added to a 384-well plate in an amount of 10. Mu.L, centrifuged, mixed, and incubated in an incubator at 23 ℃ for 90 minutes to effect the reaction.

1.5 preparation of stop mix:

a termination mixture of trypsin and a positive compound SAHA (Vorinostat) is prepared by 1x assay buffer and mixed uniformly. The reaction was stopped by adding 30. Mu.L to a 384-well plate, mixed by centrifugation, and incubated at 23 ℃ for 60 minutes in an incubator.

1.6 reading:

after terminating the reaction for 60 minutes, 384 well assay plates were placed on an Envision multifunctional microplate reader and fluorescence was read.

1.7 analytical data: the data were analyzed using XLfit5 software and IC50 values for the compounds were calculated.

TABLE 1 in vitro screening test results for the compounds of the invention

Note: a is less than or equal to 10nM; b is more than 10nM and less than or equal to 50nM; c is more than 50nM and less than or equal to 200nM; d is more than 200nM;

and (4) conclusion: the compound has the characteristics of obvious inhibition effect on HDAC6, weak inhibition on HDAC1 and high selectivity.

Experimental example 2: compound pharmacokinetic evaluation

The purpose of the experiment is as follows: test Compounds pharmacokinetics in Gongbig dogs

Experimental materials:

beagle dog (Gong, 6-15 kg, age 6 months-4, marshall Bioresources, beijing, china)

And (3) experimental operation:

beagle pharmacokinetic profiles of compounds after oral administration were tested in a standard protocol, and candidate compounds were formulated as homogeneous suspensions in the experiment and given to beagle dogs for a single oral administration. The oral solvent is polyethylene glycol-polyoxyethylene castor oil water solution with a certain proportion. Collecting a whole blood sample within 24 hours, centrifuging at 3000g for 10 minutes, separating supernatant to obtain a plasma sample, adding 30 mu L of the plasma sample into 300 mu L of acetonitrile solution containing an internal standard to precipitate protein, performing vortex centrifugation, adding 30 mu L of supernatant into equal-volume water, performing vortex centrifugation, taking the supernatant for sample injection, quantitatively analyzing the blood concentration by an LC-MS/MS analysis method, and calculating pharmacological parameters such as peak-reaching concentration, peak-reaching time, half-life period, area under a drug-hour curve and the like.

The results of the experiment are shown in table 2:

TABLE 2 pharmacokinetic test results

And (4) conclusion: the compound has good single or partial indexes of canine pharmacokinetics.

Experimental example 3: in vivo pharmacodynamic study of Combined Ixazomib administration on human myeloma MM.1S cell subcutaneous xenograft tumor CB-17 SCID model

1. Purpose of the experiment:

HDAC inhibitors are widely used in a variety of cancers and can be combined with a variety of drugs to enhance the therapeutic effect of the drugs, and are well-established anti-tumor targets. HDAC inhibitors are widely used in various cancers and can be combined with various medicaments to enhance the treatment effect of the medicaments, for example, panobinostat and proteasome inhibitor bortezomib (bortezomib) are combined to increase the curative effect of multiple myeloma and obviously reduce toxicity.

The purpose of this experiment was to investigate the efficacy of the compounds of the invention in combination with the proteasome inhibitor Ixazomib in the evaluation of the in vivo efficacy of human myeloma mm.1s cell subcutaneous xenografts in the CB-17 SCID model.

2. Experimental methods and procedures

2.1 cell culture

In vitro monolayer culture of MM.1S human myeloma cells in RPMI-1640 medium (supplier: gibco; cat # 22400-089) supplemented with 10% fetal bovine serum at 37 deg.C with 5% CO ₂ And (5) incubator culture. Passage was performed twice a week with conventional digestion treatment with pancreatin-EDTA. When the saturation degree of the cells is 80% -90%, collecting the cells, counting and inoculating.

2.2 tumor cell inoculation

0.2mL of 5X 10 ⁶ One mm.1s cell was inoculated subcutaneously on the right back (PBS: matrigel = 1: 1) of each mouse (beijing vintongli biotechnology ltd, female, 6-8 weeks old). The average tumor volume reaches 100-150mm ³ The grouped drug administration is started.

2.3 preparation of the test substances

TABLE 3 test substance preparation method

Note: the drug needs to be gently mixed well before administration to the animal.

2.4 daily Observation of Experimental animals

Animals are monitored daily for health and mortality, and routine examinations include observations of the effects of tumor growth and drug treatment on the animal's daily performance such as behavioral activity, food intake (visual only), weight changes (measured every other day), signs of appearance, or other abnormalities. The number of deaths and side effects of animals in the groups were recorded based on the number of animals in each group.

2.5 tumor measurement and Experimental indices

The experimental index is to investigate whether the tumor growth is inhibited, delayed or cured. Tumor diameters were measured three times a week with a vernier caliper. The formula for tumor volume is: v =0.5a × b ² And a and b represent the major and minor diameters of the tumor, respectively.

The antitumor therapeutic effect of the compound was evaluated by TGI (%) or relative tumor proliferation rate T/C (%). TGI (%), reflecting the rate of tumor growth inhibition. Calculation of TGI (%): TGI (%) = ((1- (average tumor volume at the end of administration of a certain treatment group-average tumor volume at the start of administration of the treatment group))/(average tumor volume at the end of treatment of solvent control group-average tumor volume at the start of treatment of solvent control group)) × 100%.

Relative tumor proliferation rate T/C (%): the calculation formula is as follows: T/C% = T _RTV /C _RTV ×100％(T _RTV : treatment group RTV; c _RTV : negative control group RTV). Calculating Relative Tumor Volume (RTV) according to the tumor measurement result, wherein the calculation formula is RTV = V _t /V ₀ In which V is ₀ When administered in groups (i.e. d) ₀ ) Measurement of the mean tumor volume, V _t Mean tumor volume at a certain measurement, T _RTV And C _RTV The same day data was taken.

After the experiment is finished, the tumor weight is detected, and T/C is calculated _weight Percent, T _weight And C _weight Tumor weights of the administered group and the vehicle control group are shown, respectively.

3. Statistical analysis

Statistical analysis, mean and Standard Error (SEM) of tumor volume for each time point for each group. Treatment groups showed the best treatment effect at day 21 after dosing at the end of the trial, and therefore statistical analysis was performed based on this data to assess differences between groups. The comparisons between two groups were analyzed using T-test, the comparisons between three or more groups were analyzed using one-way ANOVA, and if there was a significant difference in F-value, the measurements were performed using the Games-Howell method. If there is no significant difference in F value, analysis is performed using the Dunnet (2-sized) method. All data analyses were performed with SPSS 17.0. Significant differences were considered for p < 0.05.

4. Experimental results and discussion

In this experiment, the compound of example 26 was evaluated for in vivo efficacy in combination with Ixazomib in a model of human myeloma mm.1s cell xenograft tumor. Tumor volume data for this group at different time points indicate. On day 21 after the start of the administration, the tumor volume of the tumor-bearing mice in the solvent control group reached 2611mm ³ The test substance Ixazomib (4 mg/kg) alone has significant tumor inhibition effect (T/C =34.97%, TGI =68.65%, p = 0.030) compared with the solvent control group, and the tumor volume is 916mm ³ . The compound of example 26 (30 mg/kg) in combination with Ixazomib (4 mg/kg) showed significant tumor suppression (T/C =8.34%, TGI =96.88%, p = 0.001) compared to the solvent control group, with a tumor volume of 218mm ³ 。

In conclusion, the combined administration group of the compound (30 mg/kg) and Ixazomib (4 mg/kg) of example 26 in the human myeloma MM.1S cell xenograft tumor model showed the best tumor suppression effect compared with the solvent control group, and the mice showed good tolerance and no significant weight loss.

Claims

1. A compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

selected from the group consisting of: a single bond or a double bond;

n is selected from: 0 or 1;

T ₃ Selected from the group consisting of: c or N;

Z ₁ 、Z ₂ 、Z ₃ each independently selected from: CH or N;

L ₁ selected from: a single bond, -NH-, -C (= O) -NH-;

R ₁ selected from optionally substituted with 1,2 or 3R: phenyl, 6-membered heteroaryl;

R ₂ selected from: H. f, cl, br, I;

the A ring is selected from: 4-to 7-membered heterocycloalkyl;

r is selected from: F. cl, br, I;

2. A compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1, wherein R ₁ Selected from optionally substituted with 1,2 or 3R: phenyl, pyridyl.

3. A compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 2, wherein R ₁ Selected from optionally substituted with 1,2 or 3R:

4. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 3, wherein R ₁ Selected from:

5. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein ring A is selected from: oxetanyl, tetrahydrofuryl, tetrahydropyranyl, 1, 3-dioxolanyl, 1, 4-dioxacycloheptyl, 1, 4-dioxanyl, 1, 4-oxazepanyl, morpholinyl.

6. A compound, pharmaceutically acceptable salt thereof, or stereoisomer thereof according to claim 5, wherein ring A is selected from:

7. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein the structural unitSelected from:

8. a compound according to any one of claims 1 to 4, which is pharmaceutically acceptableA salt or a stereoisomer thereof, wherein the structural unitSelected from the group consisting of:

9. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to claim 8, wherein the structural unitSelected from:

10. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein the structural unitSelected from:

11. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 10, wherein the structural unitSelected from:

12. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 11, wherein the structural unitSelected from:

13. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 12, wherein the structural unitSelected from:

14. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 1 or 13, wherein the structural unitSelected from:

15. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 14, wherein the structural unitSelected from the group consisting of:

16. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein the structural unitSelected from: -CH ₂ -、-NH-、-C(＝O)-NH-、

17. The compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein the structural unitSelected from:

18. the compound, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof according to any one of claims 1 to 4, wherein the structural unitSelected from the group consisting of:

19. the compound according to any one of claims 1 to 4, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from:

wherein the content of the first and second substances,

a ring, R ₂ 、L ₁ And n is as defined in claim 1;

R ₁ as defined in any one of claims 1 to 4.

20. The compound of claim 19, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, selected from:

wherein the content of the first and second substances,

E ₁ 、E ₂ are each independently selected from-O-, -CH ₂ -and-CH ₂ -CH ₂ -；

R、R ₂ 、L ₁ And n is as defined in claim 1.

21. The following compound, or a pharmaceutically acceptable salt thereof, is selected from:

22. the compound of claim 21, or a pharmaceutically acceptable salt thereof, selected from:

23. a compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

structural unitSelected from:

n is selected from: 0 or 1;

Z ₁ 、Z ₂ 、Z ₃ each independently selected from: CH or N;

L ₁ selected from: a single bond, -NH-, -C (= O) -NH-;

R ₂ selected from: H. f, cl, br and I.

24. A compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 23, wherein the structural unitSelected from:

25. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 24, wherein the structural unitSelected from:

26. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 23, wherein the structural unitSelected from: -CH ₂ -、-NH-、-C(＝O)-NH-、

27. A compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 23, wherein the structural unitSelected from:

28. a compound, pharmaceutically acceptable salt or stereoisomer thereof according to claim 23, wherein the structural unitSelected from:

29. a pharmaceutical composition comprising as an active ingredient a therapeutically effective amount of a compound according to any one of claims 1 to 28 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

30. Use of a compound according to any one of claims 1 to 28, or a pharmaceutically acceptable salt thereof, or a composition according to claim 29, in the manufacture of a medicament for the treatment of an HDAC 6-related disorder.

31. The use of claim 30, wherein the medicament is a medicament for treating multiple myeloma.