A (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound and its preparation method and application

Abstract

The present invention discloses a (4-phenoxyphenyl) (pyrrolopyrimidine-5-yl) ketone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs, the structural formula of which is shown in Formula I, and discloses a preparation method of the above-mentioned compound and its application in preparing BTK / HDAC dual inhibitors; the activity data of the compound of the present invention show that the disclosed compound as a BTK / HDAC dual inhibitor has obvious BTK kinase and HDAC enzyme activity inhibitory effects as shown in pharmacodynamic experiments, and can be used to prepare anti-tumor drugs.

Description

Technical Field

[0001] The present invention relates to a compound, a preparation method and an application thereof, and in particular to a (4-phenoxyphenyl) (pyrrolopyrimidine-5-yl) ketone compound and a preparation method thereof, as well as an application thereof in the preparation of a drug for treating diseases caused by overexpression of BTK kinase / HDAC enzyme. Background Art

[0002] BTK (Bruton's tyrosine kinase) exists in the cytoplasm. As a member of the Tec kinase family, it is a non-receptor tyrosine kinase (NRTK). BTK plays an important role in signal transduction in the B cell receptor signaling pathway, participates in B cell activation, proliferation, differentiation and survival, and is closely related to a variety of B cell tumors and autoimmune diseases. The occurrence of cancer is related to genomic changes and epigenetic modifications, such as DNA methylation and histone modifications, which can change chromatin structure. Among them, histone modifications mainly include histone acetylation, phosphorylation and methylation. Histone acetylation is the most common epigenetic process disorder in cancer. In the cell nucleus, histone acetylation and histone deacetylation are in a dynamic equilibrium and are closely regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs). HATs transfer the acetyl group of acetyl-CoA to specific lysine residues at the amino terminus of histones. HDAC deacetylates histones, which bind tightly to negatively charged DNA, densely coil chromatin and inhibit gene transcription. In fact, these enzymes can also regulate the functions of many non-histone proteins, including nuclear transport proteins, α-tubulin in microtubules, and transcription factors. Studies have shown that HDAC can regulate cell life activities by inhibiting gene transcription, inhibiting apoptosis, regulating DNA damage processes, etc., and HDAC is overexpressed in some malignant tumors. Inhibition of HDAC can play a significant anti-cancer effect through multiple mechanisms such as inducing differentiation, cell cycle arrest, cell apoptosis and reduced angiogenesis.

[0003] In the process of clinical medication, it was found that the combination of BTK inhibitors and HDAC inhibitors can achieve better therapeutic effects. Currently, the clinical research of Ibrutinib and Abexinostat for diffuse large B-cell / mantle cell lymphoma is in Phase 1 / 2. However, combination therapy generally has disadvantages such as drug interactions, stronger toxic side effects and poor patient compliance. Summary of the invention

[0004] Objectives of the invention: The first objective of the present invention is to provide a (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound;

[0005] The second object of the present invention is to provide a method for preparing the above compound;

[0006] The third object of the present invention is the use of the above compounds as BTK / HDAC dual inhibitors.

[0007] Technical solution: The structural formula of a (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound of the present invention is as shown in Formula I:

[0008]

[0009] Wherein: at least one of W and Y exists, and W is selected from the following linking groups: ;

[0010] The Y is selected from the following linking groups: ; said n is independently selected from 1-6;

[0011] The R is selected from hydroxyl, 2-aminophenyl, 2-amino-4-fluorophenyl;

[0012] The R a , R b are independently selected from: hydrogen, halogen, cyano, nitro, hydroxyl, carboxyl, substituted or unsubstituted C 1-6 acyl, substituted or unsubstituted amino, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 one or more of the alkoxy groups;

[0013] The substituted C 1-6 Acyl, substituted amino, substituted C 1-6 Alkyl, substituted C 1-6 The substituents in the alkoxy group are selected from one or more groups: C 1-5 Alkyl, C 2-5 Alkenyl, C 2-5 Alkynyl, C 1-5 Alkoxy, halogen, nitro, cyano, hydroxy, amino, carboxyl, carbonyl, ester, amide and oxo.

[0014] Further, at least one of W and Y exists, and W is selected from the following linking groups: ;

[0015] The Y is selected from the following linking groups: ;

[0016] Each n is independently selected from 1-6;

[0017] The R is selected from hydroxyl, 2-aminophenyl, 2-amino-4-fluorophenyl;

[0018] R a , R b Independently selected from: one or more of hydrogen, halogen, cyano, nitro, hydroxyl, and carboxyl;

[0019] The halogen is selected from F, Cl, and Br.

[0020] Further, the compound is selected from 1 to 43:

[0021] .

[0022] The method for preparing the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs comprises the following steps:

[0023]

[0024] First, compound A is used as a raw material, and a halogen-metal exchange reaction is carried out with n-butyl lithium to form an organic lithium compound, and the organic lithium compound then undergoes a nucleophilic addition reaction with 4-phenoxybenzoate to obtain compound B; then a weak base is used as an acid-binding agent, and compound B undergoes a nucleophilic substitution reaction with a compound containing an amine structure to obtain a compound of formula I.

[0025] A combination drug, the active ingredient of which is selected from the (4-phenoxyphenyl) (pyrrolopyrimidine-5-yl) ketone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs.

[0026] Use of the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl)methanone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs in the preparation of drugs for treating diseases causing overexpression of BTK kinase and / or HDAC enzyme, or use of a combination drug in the preparation of drugs for treating diseases causing overexpression of BTK kinase and / or HDAC enzyme.

[0027] Use of the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl)methanone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs in the preparation of drugs for treating diseases caused by overexpression of BTK kinase and / or HDAC enzyme, or use of a combination drug in the preparation of drugs for treating diseases caused by overexpression of BTK kinase and / or HDAC enzyme.

[0028] Use of the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl)methanone compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, pharmaceutically acceptable salts or prodrugs, or combined drugs in the preparation of BTK / HDAC dual inhibitors.

[0029] Furthermore, the disease includes one or more of autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative diseases and cancer.

[0030] Further, the diseases specifically include arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjögren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, rash, anti-neutrophil cytoplasmic antibody vasculitis, pus, pus, chronic obstructive pulmonary disease, psoriasis; breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular carcinoma, glioma, endometrial cancer, melanoma, kidney cancer, bladder cancer, Cancer, bile duct cancer, pancreatic cancer, lymphoma, hairy cell cancer, nasopharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, lung cancer, small cell lung cancer, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, uterine body cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphoid leukemia, myeloid leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia.

[0031] The term "pharmaceutically acceptable" as used herein refers to any substance that does not interfere with the effectiveness of the biological activity of the active ingredient and is non-toxic to the host to which it is administered.

[0032] The pharmaceutically acceptable excipients described in the present invention are a general term for all additional materials in the drug except the main drug. The excipients should have the following properties: (1) no toxic effects on the human body and almost no side effects; (2) stable chemical properties, not easily affected by temperature, pH, storage time, etc.; (3) no incompatibility with the main drug, and no impact on the efficacy and quality inspection of the main drug; (4) no interaction with the packaging material. The excipients in the present invention include but are not limited to fillers (diluents), lubricants (glidants or anti-adhesive agents), dispersants, wetting agents, adhesives, regulators, solubilizers, antioxidants, antibacterial agents, emulsifiers, disintegrants, etc. Binders include syrup, gum arabic, gelatin, sorbitol, tragacanth gum, cellulose and its derivatives, gelatin slurry, syrup starch slurry or polyvinyl pyrrolidone, etc.; fillers include lactose, powdered sugar, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salts, sorbitol or glycine, etc.; lubricants include micropowdered silica gel, magnesium stearate, talc, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol, etc.; disintegrants include starch and its derivatives, polyvinyl pyrrolidone or microcrystalline cellulose, etc., and wetting agents include ten Sodium dialkyl sulfate, water or alcohol, etc.; antioxidants include sodium sulfite, sodium bisulfite, sodium pyrosulfite, dibutyl benzoic acid, etc.; antibacterial agents include 0.5% phenol, 0.3% cresol, 0.5% chlorobutanol, etc.; regulators include citric acid, potassium hydroxide (sodium), sodium citrate and buffers (including disodium hydrogen phosphate and sodium dihydrogen phosphate), etc.; emulsifiers include polysorbate-80, sorbitan monohydrate, Pluronic F-68, lecithin, soybean lecithin, etc.; solubilizers include Tween-80, bile, glycerol, etc. The term "pharmaceutically acceptable salt" refers to salts formed by the compounds of the present invention and acids or bases that are suitable for use as drugs. The above acids and bases are Lewis acids and bases in a broad sense. Acids suitable for forming salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, and the like; and acidic amino acids such as aspartic acid and glutamic acid.

[0033] The administration method of the compound or pharmaceutical composition of the present invention is not particularly limited, and representative administration methods include (but are not limited to): oral, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0034] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or solubilizers, for example, starches, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage form may also contain a buffer. Solid dosage forms such as tablets, pills, capsules, pills and granules may be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions may be delayed in a certain part of the digestive tract. If necessary, the active compound may also be in microencapsulated form with one or more of the above excipients.

[0035] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances. In addition to these inert diluents, the composition may also contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances. In addition to the active compound, the suspension may contain a suspending agent.

[0036] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0037] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0038] The compounds of the present invention can also be used in injection preparations. The injection is selected from liquid injection (water injection), sterile powder for injection (powder injection) or tablet for injection (referring to molded tablets or machine-pressed tablets made by aseptic operation of the drug, which are dissolved in water for injection for subcutaneous or intramuscular injection). The powder for injection contains at least an excipient in addition to the above-mentioned compound. The excipient in the present invention is a component intentionally added to the drug, which should not have pharmacological properties in the amount used, but the excipient can help the drug processing, dissolution or dissolution, targeted administration route or stability.

[0039] As described herein, the compounds of the present invention may be optionally substituted with one or more substituents, such as the general formula compounds of the present invention, or as the specific examples, subclasses, and classes of compounds included in the embodiments. It should be understood that the term "optionally substituted" can be used interchangeably with the term "substituted and unsubstituted". In general, the term "optionally", whether or not it is preceded by the term "substituted", means that one or more hydrogen atoms in the given structure are replaced by a specific substituent. Unless otherwise indicated, an optional substituted group may have a substituent substituted at each substitutable position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents may be substituted at each position the same or different. The substituents may be, but are not limited to, hydroxy, hydroxyalkyl, amino, halogen, cyano, oxo (=O), aryl, heteroaryl, alkoxy, alkyl, haloalkyl, aminoalkyl, alkylamino, alkenyl, alkynyl, heterocyclyl, thiol, nitro, aryloxy or arylalkyl, etc.

[0040] The term "alkyl" as used herein includes saturated straight or branched monovalent hydrocarbon groups of 1 to 20 carbon atoms, wherein the alkyl group may be independently optionally substituted with one or more substituents described herein. The term "haloalkane" as used herein means that the alkyl group is substituted with one or more halogen atoms which may be the same or different, wherein the alkyl group has the meaning described herein, and the halogen atom is fluorine, chlorine, bromine or iodine.

[0041] The terms "heterocycle" or "heterocyclyl" are used interchangeably herein and refer to a monocyclic, bicyclic, or tricyclic ring system in which one or more atoms in the ring may be independently and optionally replaced by heteroatoms, the ring may be fully saturated or contain one or more unsaturations, but is never aromatic, and has one or more points of attachment to other molecules. One or more hydrogen atoms in the ring may be independently and optionally replaced by one or more substituents described herein. The heterocyclyl may be a carbon group or a heteroatom group. "Heterocyclyl" also includes groups formed by fusion of a heterocyclic group with a saturated or partially unsaturated ring or heterocycle. And the heterocyclyl may be substituted or unsubstituted, wherein the substituents may be, but are not limited to, hydroxy, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkyl, alkenyl, alkynyl, heterocyclyl, thiol, nitro, aryloxy, arylalkyl, and the like.

[0042] The term "heteroatom" means one or more of O, S, N, P and Si, including N, S and P in any oxidation state; in the form of primary, secondary, tertiary amines and quaternary ammonium salts; or in the form of a nitrogen atom in a heterocyclic ring in which hydrogen is substituted.

[0043] The term "halogen" or the prefix "halo" refers to F, Cl, Br or I.

[0044] The term "aryl" can be used alone or as a part of "aralkyl", "aralkyloxy" or "aryloxyalkyl" to refer to monocyclic, bicyclic and tricyclic carbon ring systems containing 6-14 ring members, wherein at least one ring system is aromatic, wherein each ring system contains 3-7 ring members and has only one point of attachment to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring". And the aryl group can be substituted or unsubstituted, wherein the substituents can be, but are not limited to, hydroxy, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkyl, alkenyl, alkynyl, heterocyclyl, thiol, nitro, aryloxy, arylalkyl and the like.

[0045] The term "arylalkyl" means an alkyl group substituted with one or more aryl groups, wherein the alkyl and aryl groups have the meanings described herein.

[0046] The term "heteroaryl" can be used alone or as a part of "heteroarylalkyl" or "heteroarylalkoxy" to refer to monocyclic, bicyclic, and tricyclic ring systems containing 5-14 ring members, wherein at least one ring system is aromatic, and at least one ring system contains one or more heteroatoms, wherein each ring system contains 3-7 ring members, and has only one point of attachment to the rest of the molecule. The term "heteroaryl" can be used interchangeably with the term "aromatic heterocycle" or "heteroaromatic compound". And the heteroaryl group can be substituted or unsubstituted, wherein the substituents can be, but are not limited to, hydroxy, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkyl, alkenyl, alkynyl, heterocyclyl, thiol, nitro, aryloxy, arylalkyl, etc.

[0047] The term "heteroarylalkyl" means an alkyl group substituted by one or more heteroaryl groups, wherein the alkyl group and the heteroaryl group have the meanings described herein.

[0048] The term "carboxy", whether used alone or in combination with other terms, such as "carboxyalkyl", refers to -COOH.

[0049] As described in the present invention, the substituent is connected to the central ring by a bond to form a ring system (as shown in a) and represents that any substitutable position on the ring can be substituted. For example, a represents that any possible substitutable position on the piperidine ring can be substituted. .

[0050] The term "prodrug" as used in the present invention refers to a compound that is converted in vivo to a compound of formula I. Such conversion is affected by the prodrug being hydrolyzed in the blood or being converted to the parent structure by enzymes in the blood or tissues. The prodrug compounds of the present invention may be esters. In the prior invention, esters that can be used as prodrugs include phenyl esters, aliphatic (C1- 24 ) esters, acyloxymethyl esters, carbonates, carbamates and amino acid esters. For example, a compound of the present invention contains a hydroxyl group, which can be acylated to obtain a prodrug form of the compound. Other prodrug forms include phosphates, such as these phosphate compounds obtained by phosphorylation of the hydroxyl group on the parent.

[0051] Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are included within the scope of the present invention. In addition, unless otherwise indicated, the structural formulas of the compounds described in the present invention include enriched isotopes of one or more different atoms.

[0052] "Metabolite" refers to a product obtained by metabolism of a specific compound or salt thereof in vivo. Metabolites of a compound can be identified by techniques known in the art. Such products can be obtained by administering a compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, etc. Accordingly, the present invention includes metabolites of a compound, including metabolites produced by contacting a compound of the present invention with a mammal for a period of time.

[0053] The compounds of the present invention may contain asymmetric centers or chiral centers and thus exist as different stereoisomers. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, atropisomers, and mixtures thereof, such as racemic mixtures, form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefix D, L or R, S is used to indicate the absolute configuration of the chiral center of the molecule. The prefix d, l or (+), (-) is used to name the sign of rotation of the plane polarized light of the compound, the prefix (-) or l means that the compound is levorotatory, and the prefix (+) or d means that the compound is dextrorotatory. The chemical structures of these stereoisomers are the same, but their stereostructures are different. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomeric mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier.

[0054] The term "solvate" of the present invention refers to an association formed by one or more solvent molecules and the compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association formed when the solvent molecule is water. The term "protecting group" or "Pg" refers to a substituent that reacts with other functional groups, usually used to block or protect a specific functional group.

[0055] Beneficial effects: Compared with the prior art, the present invention has the following significant advantages: (4-phenoxyphenyl) (pyrrolopyrimidine-5-yl) ketone compounds have the ability to inhibit the activity of BTK and HDAC. Pharmacological experiments show that they have a significant inhibitory effect on BTK / HDAC, providing a new solution for the treatment of diseases with BTK and HDAC as therapeutic targets, such as malignant tumors or autoimmune diseases, and can be used to prepare drugs for treating related diseases, with broad application prospects. DETAILED DESCRIPTION

[0056] The present invention is further described below. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified. The present application is described in detail below in conjunction with specific embodiments.

[0057] In the present invention, the structure of the compound is determined by mass spectrometry (MS) and / or nuclear magnetic resonance ( 1 H NMR) equipment. The chemical abbreviations have the following meanings: DMF: N,N -dimethylformamide; TLC: thin layer chromatography; n-BuLi: n-butyllithium; THF: tetrahydrofuran; TEA: triethylamine; PE: petroleum ether; EA: ethyl acetate; DCM: dichloromethane; MeOH: methanol; TFA: trifluoroacetic acid; EDCI: 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride; HOBT: 1-hydroxybenzotriazole.

[0058] The compounds of the present invention can be prepared according to conventional methods in the art, and using suitable reagents, raw materials and purification methods known to those skilled in the art. The preparation method of the compounds of the present invention is described in more detail below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily carried out by a technician in the field to which the present invention belongs.

[0059] 1. Synthesis of Compounds 1 to 43

[0060] Example 1: Preparation of 4-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]butyric acid-1-hydroxyamine (Compound 1)

[0061] Step 1: Synthesis of intermediate 1-3

[0062] Compound 1-1 (10.00 g, 53.03 mmol), compound 1-2 (5.99 g, 63.63 mmol), DMF (80 mL) were added to the reaction bottle, potassium carbonate (14.66 g, 106.05 mmol) was added under stirring, and the mixture was stirred at 90°C for 3 hours. TLC showed that the reaction was complete. The reaction system was poured into water, extracted twice with ethyl acetate, the ethyl acetate layers were combined, washed twice with water, washed with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA=20:1) to obtain 12 g of colorless transparent oily liquid, which was product 1-3, with a yield of 86.15%.

[0063] Step 2: Synthesis of intermediate 1-5

[0064] Compound 1-4 (5.00 g, 21.51 mmol) was added to a three-necked reaction flask, replaced with nitrogen, and anhydrous tetrahydrofuran (20 mL) was added. The reaction solution was cooled to -78 ° C, and a tetrahydrofuran solution of n-BuLi (17.90 mL, 2.4 M) was added dropwise to the reaction flask. After stirring for 1 hour, a tetrahydrofuran solution of intermediate 1-3 (6.22 g, 23.66 mmol) was added, and the reaction was kept warm for 2 hours. TLC showed that the reaction was complete. The reaction solution was warmed to room temperature, saturated ammonium chloride solution was added to quench the reaction, ethyl acetate was added to extract twice, the ethyl acetate layers were combined, washed twice with water, washed with saturated sodium chloride solution, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (PE:EA = 5:1) to obtain 5.00 g of white solid, which was product 1-5, with a yield of 60.50%.

[0065] Step 3: Synthesis of intermediate 1-7

[0066] To the reaction flask, intermediate 1-5 (1.0 g, 2.60 mmol), compound 1-6 (536.79 mg, 5.21 mmol), anhydrous sodium carbonate (317.23 mg, 2.99 mmol) and anhydrous ethanol (30 mL) were added. The mixture was refluxed at 70 °C for 3 h. TLC showed that the reaction was complete. The mixture was concentrated under reduced pressure, water was added, and the pH was adjusted to 5-6 with glacial acetic acid. Solid precipitated, which was filtered and dried to obtain 1.15 g of white solid, which was product 1-7. The yield was 98.00%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.51 (s, 1H), 8.75 (t, J = 5.5 Hz, 1H), 8.25 (s, 1H), 7.63 (s, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.48(dd,J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.23 – 7.16 (m, 3H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 3.60 – 3.53 (m, 2H), 2.36 (t, J = 7.4 Hz,2H), 1.88 (p, J = 7.2 Hz, 2H).

[0067] Step 4: Synthesis of Intermediate 1-9

[0068] Add intermediate 1-7 (200 mg, 0.44 mmol), EDCI (170.07 mg, 0.89 mmol), HOBT (89.91 mg, 0.67 mmol), triethylamine (448.87 mg, 4.44 mmol) and DMF (5 mL) to the reaction bottle, stir at room temperature for 1 h, then add compound 1-8 (103.93 mg, 0.89 mmol), stir overnight, TLC shows that the reaction is complete. Pour the reaction system into water, extract twice with ethyl acetate, combine the ethyl acetate layers, wash twice with water, wash with saturated sodium chloride solution, dry over anhydrous Na2SO4, filter, concentrate under reduced pressure, and purify the crude product by silica gel column chromatography (DCM: MeOH = 20:1) to obtain 158.91 mg of white solid, which is product 1-9, with a yield of 65.13%.

[0069] Step 5: Synthesis of compound 1

[0070] Intermediate 1-9 (158.9 mg, 0.29 mmol) and trifluoroacetic acid (5 mL) were added to the reaction flask and stirred at room temperature for 3 h. TLC showed that the reaction was complete. The reaction solution was concentrated and chromatographed on a C18 reverse phase column to obtain 70.43 mg of a white solid, which was compound 1, with a yield of 52.32%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.75 (brs, 1H), 10.44 (s, 1H), 8.75(s, 2H), 8.25 (s, 1H), 7.63 (s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.49 (t, J =7.4 Hz, 2H), 7.26 (t, J= 7.4 Hz, 1H), 7.22 – 7.17 (m, 3H), 7.03 (dd, J =8.4, 2.4 Hz, 1H), 3.54 (q, J = 6.7 Hz, 2H), 2.11 (t, J = 7.5 Hz, 2H), 1.87(p, J = 7.4 Hz, 2H).LC / MS: m / z=466.18[M+H] + .

[0071] Example 2: Preparation of 5-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]pentan-1-hydroxyacid (Compound 2)

[0072]

[0073] Step 1: Synthesis of intermediate 2-2

[0074] Referring to the method of Example 1, intermediate 2-2 was prepared using intermediates 1-5 and 2-1 as raw materials with a yield of 80.83%. 1 HNMR (300 MHz, DMSO- d 6) δ 8.74 (t, J = 5.5 Hz, 1H), 8.24 (s, 1H), 7.61 (s,1H), 7.58 (d, J = 8.5 Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J =7.4 Hz, 1H), 7.21 – 7.17 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 3.56 (t, J = 5.8 Hz, 2H), 2.33 – 2.24 (m, 2H), 1.72 – 1.59 (m, 4H).

[0075] Step 2: Synthesis of intermediate 2-3

[0076] Referring to the method of Example 1, intermediate 2-2 and 1-8 were used as raw materials to obtain intermediate 2-3 with a yield of 41.06%.

[0077] Step 3: Synthesis of compound 2

[0078] Referring to the method of Example 1, intermediate 2-3 was used as raw material to obtain compound 2 with a yield of 76.76%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 10.39 (s, 1H), 8.78 – 8.69 (m, 2H), 8.24 (s, 1H), 7.61 (s, 1H), 7.59 (d, J =8.5Hz, 1H), 7.49(t, J =7.5Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.17 (m, 3H), 7.03 (dd, J = 8.5, 2.4 Hz,1H), 3.54 (d, J = 5.7 Hz, 2H), 2.04 (d, J = 4.7 Hz, 2H), 1.63 (s, 4H).LC / MS: m / z=480.19[M+H] + .

[0079] Example 3: Preparation N -(2-aminophenyl)-5-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl]- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]pentanamide (Compound 3)

[0080]

[0081] Referring to the method of Example 1, intermediates 2-2 and 3-1 were used as raw materials to obtain compound 3 with a yield of 30.77%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.74 (brs, 1H), 9.14 (s, 1H), 8.78 (t, J = 5.7 Hz,1H), 8.25 (s, 1H), 7.62(s, 1H),7.59 (d, J = 8.5 Hz, 1H), 7.52 – 7.44 (m,2H), 7.26 (t, J= 7.4 Hz, 1H), 7.22 – 7.12 (m, 4H), 7.03 (dd, J = 8.4, 2.4Hz, 1H), 6.89 (t, J = 8.4 Hz, 1H), 6.71 (dd, J = 8.0, 1.4 Hz, 1H), 6.57 –6.47 (m, 1H), 4.88 (s, 2H), 3.59 (d, J = 5.9 Hz, 2H), 2.39 (d, J = 6.9 Hz, 2H), 1.80 – 1.66 (m, 4H).LC / MS: m / z=555.16[M+H] + .

[0082] Example 4: Preparation N -(2-amino-4-fluorophenyl)-5-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl]- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]pentanamide (Compound 4)

[0083]

[0084] Referring to the method of Example 1, intermediates 2-2 and 4-1 were used as raw materials to prepare compound 4 with a yield of 36.81%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.71 (brs, 1H), 9.05 (s, 1H), 8.77 (s, 1H), 8.25(s, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.4 Hz, 1H), 7.48 (t, J = 7.8 Hz, 2H),7.26 (t, J = 7.3 Hz, 1H), 7.20 - 7.18 (m, 3H), 7.10 (dd, J = 8.7, 6.3 Hz,1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 6.48 (dd, J = 11.2, 2.9 Hz, 1H), 6.31 -6.26 (m, 1H), 5.14 (s, 2H), 3.59 (d,J = 6.0 Hz, 2H), 2.39 (t, J = 5.2 Hz, 2H), 1.73 (s, 4H).LC / MS: m / z=573.14[M+H] + .

[0085] Example 5: Preparation of 6-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]hexane-1-hydroxyamine (Compound 5)

[0086]

[0087] Step 1: Synthesis of intermediate 5-2

[0088] Referring to the method of Example 1, intermediates 1-5 and 5-1 were used as raw materials to prepare compound 5-2 with a yield of 92.26%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.30 (s, 1H), 8.74 (t, J = 5.5 Hz, 1H), 8.24 (s,1H), 7.61 (s, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.52 – 7.44 (m, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 – 7.17 (m, 3H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 3.54 (q, J = 6.6 Hz, 2H), 2.23 (t, J = 7.3 Hz, 2H), 1.73 – 1.54 (m, 4H), 1.43 (q, J =8.2 Hz, 2H).

[0089] Step 2: Synthesis of intermediate 5-3

[0090] Referring to the method of Example 1, intermediates 5-2 and 1-8 were used as raw materials to prepare compound 5-3 with a yield of 45.24%.

[0091] Step 3: Synthesis of compound 5

[0092] Referring to the method of Example 1, intermediate 5-3 was used as raw material to obtain compound 5 with a yield of 49.00%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.75 (brs, 1H), 10.36 (s, 1H), 8.90 – 8.59 (m, 2H), 8.25 (s, 1H), 7.62 (s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.49 (t, J = 8.5 Hz,2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.16 (m, 3H), 7.03 (dd, J = 8.5, 2.4Hz, 1H), 3.53 (q, J = 6.4 Hz, 2H), 1.97 (t, J = 7.3 Hz, 2H), 1.72 – 1.52 (m,4H), 1.40 (q, J =7.9 Hz, 2H).LC / MS: m / z=494.20[M+H] + .

[0093] Example 6: Preparation N -(2-aminophenyl)-6-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]hexanamide (Compound 6)

[0094]

[0095] Referring to the method of Example 1, intermediates 5-2 and 3-1 were used as raw materials to obtain compound 6 with a yield of 47.37%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.26 (brs, 1H),9.07 (s, 1H), 8.76 (t, J = 5.4 Hz,1H), 8.24 (s, 1H), 7.60 (s, 1H), 7.57 (d, J = 8.4 Hz, 1H), 7.52 – 7.46 (m,2H), 7.26 (t, J= 7.4 Hz, 1H), 7.22 – 7.17 (m, 3H), 7.14 (dd, J = 7.9, 1.5Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.91 – 6.85 (m, 1H), 6.70 (dd, J =8.0, 1.5 Hz, 1H), 6.53 – 6.47 (m, 1H), 4.82 (s, 2H), 3.56 (q, J = 6.8 Hz, 2H), 2.35 (t, J = 7.4 Hz, 2H), 1.75 – 1.63 (m, 4H), 1.53–1.48 (m, 2H).LC / MS: m / z=569.17[M+H] + .

[0096] Example 7: Preparation N -(2-amino-4-fluorophenyl)-6-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]hexanamide (Compound 7)

[0097]

[0098] Referring to the method of Example 1, intermediates 5-2 and 4-1 were used as raw materials to obtain compound 7 with a yield of 59.77%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.70 (brs, 1H), 9.01 (s, 1H), 8.75 (t, J = 5.5 Hz,1H), 8.25 (s, 1H), 7.60 (s, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.48 (t, J = 7.8Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.20 – 7.18 (m, 3H), 7.08 (dd, J = 8.7,6.3 Hz, 1H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 6.48 (dd, J= 11.2, 2.9 Hz, 1H), 6.27 – 6.22 (m, 1H), 5.12 (s, 2H), 3.57 (q, J = 6.6 Hz, 2H), 2.33 (t, J = 7.5Hz, 2H), 1.71 – 1.65 (m, 4H), 1.48 (q, J = 8.2 Hz, 2H).LC / MS: m / z=587.27[M+H] + .

[0099] Example 8: Preparation of 7-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]heptyl-1-hydroxyacid (Compound 8)

[0100]

[0101] Step 1: Synthesis of intermediate 8-2

[0102] Referring to the method of Example 1, intermediates 1-5 and 8-1 were used as raw materials to prepare compound 8-2 with a yield of 85.56%. 1 HNMR (300 MHz, DMSO- d 6) δ 8.75 (t, J = 5.5 Hz, 1H), 8.23 ​​(s, 1H), 7.58 (d, J =8.9 Hz, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.25 (t, J = 7.4 Hz, 1H), 7.22 –7.15 (m, 3H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 3.53 (q, J = 6.5 Hz, 2H), 2.18(t, J = 7.3 Hz, 2H), 1.68 – 1.59 (m, 2H), 1.55 – 1.46 (m, 2H), 1.45 – 1.30 (m, 4H).

[0103] Step 2: Synthesis of intermediate 8-3

[0104] Referring to the method of Example 1, intermediates 8-2 and 1-8 were used as raw materials to prepare compound 8-3 with a yield of 57.24%.

[0105] Step 3: Synthesis of compound 8

[0106] Referring to the method of Example 1, compound 8 was prepared using intermediate 8-3 as raw material with a yield of 50.30%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 10.35 (s, 1H), 8.77 (s, 1H), 8.68 (s,1H), 8.24 (s, 1H), 7.61 (s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.48 (t, J = 7.9Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.20-7.18 (m, 3H), 7.03 (dd, J = 8.4, 2.4Hz, 1H), 3.53 (q, J = 6.6 Hz, 2H), 1.95 (t, J = 7.3 Hz, 2H), 1.69 – 1.59 (m,2H), 1.56 – 1.46 (m, 2H), 1.44 – 1.30 (m, 4H).LC / MS: m / z=508.21[M+H] + .

[0107] Example 9: Preparation N -(2-aminophenyl)-7-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]heptylamide (Compound 9)

[0108]

[0109] Referring to the method of Example 1, intermediates 8-2 and 3-1 were used as raw materials to prepare compound 9 with a yield of 49.82%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.67 (brs, 1H), 9.10 (s, 1H), 8.76 (t, J = 5.6 Hz,,1H), 8.25 (s, 1H), 7.61 (s, 1H), 7.58 (d, J= 8.5 Hz, 1H), 7.48 (t, J = 7.9Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.20– 7.18 (m, 3H), 7.14 (d, J = 6.9 Hz,1H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 6.88 (t, J = 7.3 Hz, 1H), 6.71 (d, J =6.6 Hz, 1H), 6.52 (t, J = 7.6 Hz, 1H), 4.81 (s, 2H), 3.56 (q, J =5.8 Hz, 2H),2.32 (t, J = 7.3 Hz, 2H), 1.70 – 1.60 (m, 4H), 1.51 – 1.37 (m, 4H).LC / MS: m / z=583.18[M+H] + .

[0110] Example 10: Preparation N -(2-amino-4-fluorophenyl)-7-[(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino]heptylamide (Compound 10)

[0111]

[0112] Referring to the method of Example 1, intermediates 8-2 and 4-1 were used as raw materials to prepare compound 10 with a yield of 27.84%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.70 (brs, 1H), 9.01 (s, 1H), 8.75 (t, J = 5.4 Hz,1H), 8.25 (s, 1H), 7.60 (s, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.48 (t, J = 7.9Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.20– 7.18 (dd, J= 5.3, 2.8 Hz, 3H), 7.08(dd, J = 8.7, 6.3 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 6.50 – 6.46 (m,1H), 6.31 – 6.26 (m, 1H), 5.12 (s, 2H), 3.56 (q, J = 6.5 Hz, 2H), 2.31 (t, J = 7.5 Hz, 2H), 1.69 – 1.60(m, 4H), 1.49 – 1.37 (m, 4H).LC / MS: m / z=601.34[M+H] + .

[0113] Example 11: Preparation of [1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]methanehydroxyamine (Compound 11)

[0114]

[0115] Step 1: Synthesis of intermediate 11-2

[0116] Referring to the method of Example 1, intermediate 11-2 was prepared using intermediates 1-5 and 11-1 as raw materials, with a yield of 81.05%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.40 (s, 1H), 8.32 (s, 1H), 7.63 – 7.59 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.15 (m,3H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 4.17 – 4.12 (m, 2H), 3.04 (t, J = 10.9Hz, 2H), 2.49 – 2.42 (m, 1H), 1.85 – 1.80 (m, 2H), 1.72 – 1.64 (m, 2H).

[0117] Step 2: Synthesis of Intermediate 11-3

[0118] Referring to the method of Example 1, intermediate 11-2 and 1-8 were used as raw materials to obtain intermediate 11-3 with a yield of 60.74%.

[0119] Step 3: Synthesis of compound 11

[0120] Referring to the method of Example 1, compound 11 was prepared using intermediate 11-3 as raw material with a yield of 54.17%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.66 (brs, 1H), 10.46 (s, 1H), 8.72 (brs, 1H), 8.32 (s,1H), 7.65 – 7.58 (m, 2H), 7.53 – 7.43 (m, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.23– 7.15 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 4.27 (d, J = 12.9 Hz, 2H),3.02 – 2.86 (m, 2H), 2.34 – 2.16 (m, 1H), 1.81 – 1.52 (m, 4H).LC / MS: m / z=492.09[M+H] + .

[0121] Example 12: Preparation N -(2-aminophenyl)-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 12)

[0122]

[0123] Referring to the method of Example 1, intermediates 11-2 and 3-1 were used as raw materials to prepare compound 12 with a yield of 46.56%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.65 (brs, 1H), 9.13 (s, 1H), 8.34 (s, 1H), 7.64 –7.61 (m, 2H), 7.48 (t, J = 7.8 Hz, 2H), 7.26 (t, J= 7.4 Hz, 1H), 7.22 – 7.11(m, 4H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 6.89 (t, J = 7.5 Hz, 1H), 6.71 (d, J = 7.7 Hz, 1H), 6.53 (t, J = 7.6 Hz, 1H), 4.81 (s, 2H), 4.32 (d, J = 12.3 Hz,2H), 3.04 – 2.97 (m, 2H), 2.71 – 2.61 (m, 1H), 1.89 – 1.64 (m, 4H).LC / MS: m / z=567.19[M+H] + .

[0124] Example 13: Preparation N -(2-amino-4-fluorophenyl)-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 13)

[0125]

[0126] Referring to the method of Example 1, intermediates 11-2 and 4-1 were used as raw materials to prepare compound 13 with a yield of 45.14%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.63 (brs, 1H), 9.04 (s, 1H), 8.33 (s, 1H), 7.62(d, J = 8.2 Hz, 2H), 7.48 (dd, J = 8.6, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz,1H), 7.22 – 7.15 (m, 3H), 7.09 (dd, J = 8.7, 6.3 Hz, 1H), 7.03 (dd, J = 8.5,2.4 Hz, 1H), 6.49 (dd, J = 11.2, 2.9 Hz, 1H), 6.32 – 6.27 (m, 1H), 5.11 (s,2H), 4.31 (d,J = 12.6 Hz, 2H), 3.00 (t, J = 11.1 Hz, 2H), 2.68 – 2.59 (m,1H), 1.87 – 1.73 (m, 4H).LC / MS: m / z=585.28[M+H] + .

[0127] Example 14: Preparation of 4-({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]carbonyl}amino)butyric acid-1-hydroxyamine (Compound 14)

[0128]

[0129] Step 1: Synthesis of intermediate 14-2

[0130] Referring to the method of Example 1, intermediates 11-2 and 14-1 were used as raw materials to obtain compound 14-2 with a yield of 60.44%. LC / MS: m / z=576.33[M+H] + .

[0131] Step 2: Synthesis of compound 14-3

[0132] Add intermediate 14-2 (654.00 mg, 1.14 mmol), NaOH (90.82 mg, 2.27 mmol), methanol (8 mL) and water (2 mL) to the reaction flask, reflux at 40 °C for 3 h, TLC shows that the reaction is complete. Concentrate, add 50 mL of water, add acetic acid dropwise until the pH value of the solution is 5-6, a white solid precipitates, filter and dry to obtain intermediate 14-3 (566.90 mg), yield: 88.85%. 1 H NMR (400 MHz, DMSO- d 6) δ 8.30 (s, 1H), 7.88 (t, J = 5.5 Hz, 1H),7.60 (d, J = 8.4 Hz, 1H), 7.58 (s, 1H), 7.48 (dd, J = 8.5, 7.4 Hz, 2H), 7.25(t, J = 7.4 Hz, 1H), 7.21 – 7.17 (m, 2H), 7.15 (d, J = 2.4 Hz, 1H), 7.02 (dd,J = 8.5, 2.4 Hz, 1H), 4.25 (d, J = 13.1 Hz, 2H), 3.03 (q, J = 6.5 Hz, 2H),2.97 – 2.89 (m, 2H), 2.38 – 2.31 (m, 1H), 2.14 (t, J = 7.3 Hz, 2H), 1.69 –1.64 (m, 4H), 1.59 (t, J = 7.1 Hz, 2H). LC / MS: m / z=562.20[M+H] + .

[0133] Step 3: Synthesis of compound 14-4

[0134] Referring to the method of Example 1, intermediate 14-3 and 1-8 were used as raw materials to obtain intermediate 14-4 with a yield of 77.80%. LC / MS: m / z=661.42[M+H] + .

[0135] Step 4: Synthesis of compound 14

[0136] Referring to the method of Example 1, intermediate 14-4 was used as raw material to obtain compound 14 with a yield of 73.43%. 1 H NMR (400 MHz, DMSO- d 6) δ 12.64 (brs, 1H), 10.36 (s, 1H), 8.70 (s, 1H), 8.32 (s,1H), 7.81 (t, J = 5.6 Hz, 1H), 7.61 (d, J = 5.8 Hz, 1H), 7.60 (s, 1H), 7.48(dd, J = 8.6, 7.3 Hz, 2H), 7.26 (t, J = 7.5 Hz, 1H), 7.19 (d, J = 7.4 Hz,2H), 7.16 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 4.26 (d, J =13.0 Hz, 2H), 3.01 (q, J= 6.8 Hz, 2H), 2.96 – 2.91 (m, 2H), 2.39 – 2.31 (m,1H), 1.93 (t, J = 7.6 Hz, 2H), 1.73 – 1.64 (m, 4H), 1.63 – 1.56 (m, 2H).LC / MS: m / z=577.29[M+H] + .

[0137] Example 15: Preparation N -{4-[(2-aminophenyl)amino]-4-oxydebutylene}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 15)

[0138]

[0139] Referring to the method of Example 1, intermediates 14-3 and 3-1 were used as raw materials to prepare compound 15 with a yield of 44.04%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.64 (brs, 1H), 9.09 (s, 1H), 8.32 (s, 1H), 7.86(t, J = 5.7 Hz, 1H), 7.65 – 7.57 (m, 2H), 7.48 (dd, J = 8.6, 7.3 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.20 (d, J = 1.3 Hz, 1H), 7.18 (d, J = 1.0 Hz, 1H),7.17 (d, J = 2.4 Hz, 1H), 7.13 (dd, J = 7.9, 1.5 Hz, 1H), 7.03 (dd, J = 8.5,2.4 Hz, 1H), 6.88 (td, J = 7.6, 1.5 Hz, 1H), 6.70 (dd, J = 8.0, 1.5 Hz, 1H), 6.54 – 6.50 (m, 1H), 4.86 (s, 2H), 4.27 (d,J = 13.1 Hz, 2H), 3.12 – 3.07 (m,2H), 3.01 – 2.89 (m, 2H), 2.41 – 2.33 (m, 1H), 2.30 (t, J = 7.4 Hz, 2H), 1.74– 1.59 (m, 6H).LC / MS: m / z=652.40[M+H] + .

[0140] Example 16: Preparation N -{4-[(2-amino-4-fluorophenyl)amino]-4-oxydebutylene}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 16)

[0141]

[0142] Referring to the method of Example 1, intermediates 14-3 and 4-1 were used as raw materials to prepare compound 16 with a yield of 31.28%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.64 (brs, 1H), 9.01 (s, 1H), 8.32 (s, 1H), 7.85(t, J = 5.7 Hz, 1H), 7.61 (t, J = 4.3 Hz, 2H), 7.48 (dd, J = 8.5, 7.3 Hz,2H), 7.26 (t, J = 7.4 Hz, 1H), 7.19 (d, J = 7.4 Hz, 2H), 7.16 (d, J = 2.4 Hz,1H), 7.07 (dd, J = 8.7, 6.3 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 6.47(dd, J = 11.2, 2.9 Hz, 1H), 6.31 – 6.26 (m, 1H), 5.19 (s, 2H), 4.27 (d, J=13.0 Hz, 2H), 3.11 – 3.06 (m, 2H), 2.98 – 2.92 (m, 2H), 2.41 – 2.33 (m, 1H), 2.28 (t, J = 7.4 Hz, 2H), 1.73 – 1.66 (m, 6H).LC / MS: m / z=670.31[M+H] + .

[0143] Example 17: Preparation of 5-({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]carbonyl}amino)pentan-1-hydroxyamine (Compound 17)

[0144]

[0145] Step 1: Synthesis of Intermediate 17-2

[0146] Referring to the method of Example 1, intermediates 11-2 and 17-1 were used as raw materials to obtain compound 17-2 with a yield of 83.62%. LC / MS: m / z=590.34[M+H] + .

[0147] Step 2: Synthesis of Intermediate 17-3

[0148] Referring to the method of Example 14, compound 17-3 was prepared using intermediate 17-2 as raw material with a yield of 95.96%. 1 HNMR (400 MHz, DMSO- d 6) δ 7.79 (t, J = 5.7 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H),7.58 (s, 1H), 7.48 (dd, J = 8.6, 7.4 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21– 7.17 (m, 2H), 7.15 (d, J = 2.4 Hz, 1H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H),4.26 (d, J = 13.3 Hz, 2H), 3.01 (q, J= 6.4 Hz, 2H), 2.97 – 2.90 (m, 2H), 2.39 – 2.31 (m, 1H), 2.15 (t, J = 7.2 Hz, 2H), 1.72 – 1.62 (m, 4H), 1.49 –1.35 (m, 4H). LC / MS: m / z=576.21[M+H] + .

[0149] Step 3: Synthesis of compound 17-4

[0150] Referring to the method of Example 1, intermediate 17-3 and compound 1-8 were used as raw materials to obtain compound 17-4 with a yield of 68.44%. LC / MS: m / z=675.37[M+H] + .

[0151] Step 4: Synthesis of compound 17

[0152] Referring to the method of Example 1, intermediate 17-4 was used as raw material to prepare compound 17 with a yield of 71.36%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.64 (brs, 1H), 10.37 (s, 1H), 8.68 (s, 1H), 8.31 (s,1H), 7.79 (t, J = 5.6 Hz, 1H), 7.63 – 7.58 (m, 2H), 7.48 (dd, J = 8.5, 7.3Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.16 (m, 3H), 7.03 (dd, J = 8.5,2.4 Hz, 1H), 4.26 (d, J = 12.8 Hz, 2H), 3.06 – 2.89 (m, 4H), 2.40 – 2.30 (m,1H), 1.93 (t, J = 7.1 Hz, 2H), 1.70 – 1.61 (s, 4H), 1.51 – 1.36 (m, 4H).LC / MS: m / z=591.30[M+H] + .

[0153] Example 18: Preparation N-{5-[(2-aminophenyl)amino]-5-oxypentylene}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 18)

[0154]

[0155] Referring to the method of Example 1, intermediates 17-3 and 3-1 were used as raw materials to prepare compound 18 with a yield of 27.07%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.63 (brs, 1H), 9.09 (s, 1H), 8.31 (s, 1H), 7.82(t, J = 5.7 Hz, 1H), 7.63 – 7.58 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.13 (m, 4H), 7.02 (dd, J = 8.5, 2.4 Hz,1H), 6.91 – 6.85 (m, 1H), 6.72 – 6.69 (m, 1H), 6.55 – 6.49 (m, 1H), 4.82 (s,2H), 4.26 (d, J = 12.4 Hz, 2H), 3.06 (q, J = 6.6 Hz, 2H), 3.01 – 2.87 (m,2H), 2.41 – 2.28(m, 3H), 1.67 (s, 4H), 1.60 – 1.40 (m, 4H).LC / MS: m / z=666.35[M+H] + .

[0156] Example 19: Preparation N -{5-[(2-amino-4-fluorophenyl)amino]-5-oxopentyl}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 19)

[0157]

[0158] Referring to the method of Example 1, intermediates 17-3 and 4-1 were used as raw materials to prepare compound 19 with a yield of 22.48%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.62 (brs, 1H), 9.02 (s, 1H), 8.31 (s, 1H), 7.82(t, J = 5.6 Hz, 1H), 7.62 – 7.59 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.15 (m, 3H), 7.08 (dd, J = 8.7, 6.3 Hz,1H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 6.50 – 6.45 (m, 1H), 6.32 – 6.26 (m,1H), 5.14 (s, 2H), 4.26 (d, J = 12.3 Hz, 2H), 3.08 – 2.89 (m, 4H), 2.41 –2.26 (m, 3H), 1.71 – 1.62 (m, 4H), 1.61 – 1.51 (m, 2H), 1.47 – 1.39 (m, 2H).LC / MS: m / z=684.32[M+H] + .

[0159] Example 20: Preparation of 6-({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]carbonyl}amino)hexane-1-hydroxyamine (Compound 20)

[0160]

[0161] Step 1: Synthesis of intermediate 20-2

[0162] Referring to the method of Example 1, intermediates 11-2 and 20-1 were used as raw materials to obtain compound 20-2 with a yield of 52.89%. LC / MS: m / z=604.35[M+H] + .

[0163] Step 2: Synthesis of intermediate 20-3

[0164] Referring to the method of Example 14, compound 20-3 was prepared using intermediate 20-2 as raw material. The yield was 89.96%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.31 (s, 1H), 7.75 (t, J = 5.6 Hz, 1H), 7.62 – 7.58(m, 2H), 7.48 (dd, J = 8.6, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 –7.17 (m, 2H), 7.16 (d, J = 2.4 Hz, 1H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 4.26(d, J = 13.1 Hz, 2H), 3.03 – 2.98 (m, 2H), 2.95 – 2.90 (m, 2H), 2.39 – 2.31(m, 1H), 2.17 (t, J = 7.3 Hz, 2H), 1.69 – 1.63 (m, 4H), 1.50 – 1.45 (m, 2H), 1.40 – 1.34 (m, 2H), 1.24 (q, J = 5.2, 4.2 Hz, 2H). LC / MS: m / z=590.22[M+H] + .

[0165] Step 3: Synthesis of intermediate 20-4

[0166] Referring to the method of Example 1, intermediate 20-3 and 1-8 were used as raw materials to obtain intermediate 20-4, with a yield of 44.46%. LC / MS: m / z=689.38[M+H] + .

[0167] Step 4: Synthesis of compound 20

[0168] Referring to the method of Example 1, intermediate 20-4 was used as raw material to prepare compound 20 with a yield of 63.01%. 1 H NMR (400 MHz, DMSO- d6) δ 12.64 (brs, 1H), 10.34 (s, 1H), 8.68 (s, 1H), 8.31 (s,1H), 7.77 (t, J = 5.6 Hz, 1H), 7.61 (d, J = 6.3 Hz, 1H), 7.60 (s, 1H), 7.48(dd, J = 8.6, 7.4 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 – 7.18 (m, 2H), 7.16 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 4.26 (d, J = 13.0Hz, 2H), 3.00 (q, J = 6.5 Hz, 2H), 2.97 – 2.89 (m, 2H), 2.38 – 2.31 (m, 1H),1.92 (t, J = 7.4 Hz, 2H), 1.71 – 1.63 (m, 4H), 1.51 – 1.43 (m, 2H), 1.40 –1.33 (m, 2H), 1.24 – 1.16 (m, 2H).LC / MS: m / z=605.31[M+H] + .

[0169] Example 21: Preparation N -{6-[(2-aminophenyl)amino]-6-oxyhexylene}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 21)

[0170]

[0171] Referring to the method of Example 1, intermediates 20-3 and 3-1 were used as raw materials to prepare compound 21 with a yield of 58.21%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.63 (brs, 1H), 9.09 (s, 1H), 8.31 (s, 1H), 7.79(t, J= 5.6 Hz, 1H), 7.63 – 7.57 (m, 2H), 7.48 (dd, J = 8.6, 7.4 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.17 (m, 2H), 7.16 (d, J = 2.4 Hz, 1H), 7.14(dd, J = 7.9, 1.5 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.90 – 6.85 (m, 1H), 6.72 – 6.69 (m, 1H), 6.54 – 6.50 (m, 1H), 4.81 (s, 2H), 4.26 (d, J =13.0 Hz, 2H), 3.03 (q, J = 6.3 Hz, 2H), 2.97 – 2.90 (m, 2H), 2.39 – 2.33 (m,1H), 2.29 (t, J = 7.4 Hz, 2H), 1.70 – 1.64 (m, 4H), 1.62 – 1.54 (m, 2H), 1.45 – 1.38 (m, 2H), 1.33 – 1.27 (m, 2H).LC / MS: m / z=680.36[M+H] + .

[0172] Example 22: Preparation N -{6-[(2-amino-4-fluorophenyl)amino]-6-oxyhexylene}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 22)

[0173]

[0174] Referring to the method of Example 1, intermediates 20-3 and 4-1 were used as raw materials to prepare compound 22 with a yield of 68.20%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.64 (brs, 1H), 9.01 (s, 1H), 8.31 (s, 1H), 7.79(t, J= 5.6 Hz, 1H), 7.62 – 7.58 (m, 2H), 7.48 (dd, J = 8.6, 7.3 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.17 (m, 2H), 7.16 (d, J = 2.4 Hz, 1H), 7.08(dd, J = 8.7, 6.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.49 – 6.46 (m,1H), 6.31 – 6.26 (m, 1H), 5.13 (s, 2H), 4.26 (d, J = 13.0 Hz, 2H), 3.03 (q, J = 6.5 Hz, 2H), 2.98 – 2.89 (m, 2H), 2.37 – 2.31 (m, 1H), 2.28 (t, J = 7.4 Hz,2H), 1.70 – 1.64 (m, 4H), 1.61 – 1.53 (m, 2H), 1.45 – 1.37 (m, 2H), 1.32 –1.24 (m, 2H).LC / MS: m / z=698.40[M+H] + .

[0175] Example 23: Preparation of 7-({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]carbonyl}amino)heptyl-1-hydroxyamine (Compound 23)

[0176]

[0177] Step 1: Synthesis of intermediate 23-2

[0178] Referring to the method of Example 1, intermediates 11-2 and 23-1 were used as raw materials to obtain compound 23-2 with a yield of 54.30%. LC / MS: m / z=618.55[M+H] + .

[0179] Step 2: Synthesis of intermediate 23-3

[0180] Referring to the method of Example 14, compound 23-3 was prepared using intermediate 23-2 as raw material. The yield was 100%. 1 H NMR (400 MHz, DMSO- d 6) δ 12.65 (s, 1H), 8.31 (s, 1H), 7.74 (t, J = 5.6 Hz, 1H),7.62 – 7.58 (m, 2H), 7.48 (t, J = 7.9 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.19(d, J = 8.0 Hz, 2H), 7.16 (d, J = 2.3 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz,1H), 4.26 (d, J = 13.0 Hz, 2H), 3.03 – 2.98 (m, 2H), 2.97 – 2.90 (m, 2H), 2.39 – 2.31 (m, 1H), 2.18 (t, J = 7.3 Hz, 2H), 1.69 – 1.64 (m, 4H), 1.47 (t, J = 7.1 Hz, 2H), 1.36 (t, J = 6.8 Hz, 2H), 1.26 – 1.22 (m, 4H). LC / MS: m / z=604.23[M+H] + .

[0181] Step 3: Synthesis of Intermediate 23-4

[0182] Referring to the method of Example 1, intermediate 23-3 and 1-8 were used as raw materials to obtain intermediate 23-4 with a yield of 67.21%. LC / MS: m / z=703.52[M+H] + .

[0183] Step 4: Synthesis of compound 23

[0184] Referring to the method of Example 1, intermediate 23-4 was used as raw material to prepare compound 23 with a yield of 84.75%. 1 H NMR (400 MHz, DMSO- d6) δ 12.65 (brs, 1H), 10.34 (s, 1H), 8.68 (brs, 1H), 8.32 (s,1H), 7.76 (t, J = 5.7 Hz, 1H), 7.61 (d, J = 5.8 Hz, 1H), 7.60 (s, 1H), 7.48(dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 – 7.17 (m, 2H), 7.16 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 4.26 (d, J = 12.9Hz, 2H), 3.02 – 2.97 (m, 2H), 2.96 – 2.91 (m, 2H), 2.40 – 2.30 (m, 1H), 1.92(t, J = 7.4 Hz, 2H), 1.69 – 1.64 (m, 4H), 1.50 – 1.42 (m, 2H), 1.37 – 1.32(m, 2H), 1.26 – 1.18 (m, 4H).LC / MS: m / z=619.38[M+H] + .

[0185] Example 24: Preparation N -{7-[(2-aminophenyl)amino]-7-oxyheptanyl}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 24)

[0186]

[0187] Referring to the method of Example 1, intermediates 23-3 and 3-1 were used as raw materials to prepare compound 24 with a yield of 60.48%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.63 (brs, 1H), 9.11 (s, 1H), 8.32 (s, 1H), 7.77(t, J= 5.6 Hz, 1H), 7.63 – 7.59 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H),7.26 (t, J = 7.4 Hz, 1H), 7.19 (dd, J = 7.4, 1.6 Hz, 2H), 7.16 (d, J = 2.4Hz, 1H), 7.14 (dd, J = 7.9, 1.5 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.91– 6.87 (m, 1H), 6.73 – 6.70 (m, 1H), 6.56 – 6.52 (m, 1H), 4.89 (brs, 2H), 4.26 (d, J = 13.0 Hz, 2H), 3.02 (q, J = 6.5 Hz, 2H), 2.97 – 2.90 (m, 2H), 2.39 – 2.33 (m, 1H), 2.30 (t, J = 7.4 Hz, 2H), 1.73 – 1.63 (m, 4H), 1.61 –1.54 (m, 2H), 1.43 – 1.36 (m, 2H), 1.33 – 1.27 (m, 4H).LC / MS: m / z=694.50[M+H] + .

[0188] Example 25: Preparation N -{7-[(2-amino-4-fluorophenyl)amino]-7-oxyheptanyl}-1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridine-4-carboxamide (Compound 25)

[0189]

[0190] Referring to the method of Example 1, intermediates 23-3 and 4-1 were used as raw materials to prepare compound 25 with a yield of 59.37%. 1 HNMR (400 MHz, DMSO- d6) δ 12.64 (brs, 1H), 9.02 (s, 1H), 8.32 (s, 1H), 7.77(t, J = 5.6 Hz, 1H), 7.61 (d, J = 5.8 Hz, 1H), 7.60 (s, 1H), 7.48 (dd, J =8.6, 7.3 Hz, 2H), 7.27 (d, J = 7.4 Hz, 1H), 7.21 – 7.17 (m, 2H), 7.16 (d, J =2.4 Hz, 1H), 7.08 (dd, J = 8.7, 6.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.50 – 6.46 (m, 1H), 6.32 – 6.27 (m, 1H), 5.13 (brs, 2H), 4.26 (d, J = 13.1Hz, 2H), 3.02 (q, J = 6.6 Hz, 2H), 2.97 – 2.90 (m, 2H), 2.39 – 2.32 (m, 1H), 2.28 (t, J = 7.4 Hz, 2H), 1.71 – 1.64 (m, 4H), 1.60 – 1.53 (m, 2H), 1.42 –1.35 (m, 2H), 1.33 – 1.23 (m, 4H). LC / MS: m / z=712.47[M+H] + .

[0191] Example 26: Preparation of {4-[({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)hexahydropyridin-4-yl]carbonyl}amino)methyl]phenyl}methanehydroxylamine (Compound 26)

[0192]

[0193] Step 1: Synthesis of intermediate 26-2

[0194] Referring to the method of Example 1, intermediates 11-2 and 26-1 were used as raw materials to obtain compound 26-2 with a yield of 40.04%. LC / MS: m / z=624.44[M+H] + .

[0195] Step 2: Synthesis of intermediate 26-3

[0196] Referring to the method of Example 14, compound 26-3 was prepared using intermediate 26-2 as raw material with a yield of 97.61%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.64 (s, 1H), 8.41 (t, J = 6.0 Hz, 1H), 8.32 (s,1H), 7.88 (d, J = 8.3 Hz, 2H), 7.61 (d, J = 8.8 Hz, 2H), 7.48 (d, J = 15.9Hz, 2H), 7.33 (d, J = 8.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 – 7.18 (m,2H), 7.16 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 4.33 (d, J =5.9 Hz, 2H), 4.28 (d, J = 13.4 Hz, 2H), 3.01 – 2.94 (m, 2H), 2.49 – 2.44 (m,1H), 1.77 – 1.70 (m, 4H).LC / MS: m / z=610.30[M+H] + .

[0197] Step 3: Synthesis of compound 26-4

[0198] Referring to the method of Example 1, intermediate 26-3 and 1-8 were used as raw materials to obtain intermediate 26-4 with a yield of 49.44%. LC / MS: m / z=709.34[M+H] + .

[0199] Step 4: Synthesis of compound 26

[0200] Referring to the method of Example 1, compound 26 was prepared using intermediate 26-4 as raw material. The yield was 44.42%. 1 H NMR (400 MHz, DMSO- d 6) δ 12.65 (brs, 1H), 11.17 (s, 1H), 9.01 (s, 1H), 8.41 (t, J = 5.9 Hz, 1H), 8.32 (s, 1H), 7.69 (d, J = 8.3 Hz, 2H), 7.63 – 7.59 (m, 2H), 7.48 (dd, J = 8.5, 7.4 Hz, 2H), 7.31 – 7.24 (m, 3H), 7.20 (dd, J = 8.6, 1.1Hz, 2H), 7.17 (d, J = 2.4 Hz, 1H), 7.03 (dd, J = 8.4, 2.4 Hz, 1H), 4.30 –4.26 (m, 4H), 3.01 – 2.91 (m, 2H), 2.48 – 2.43 (m, 1H), 1.78 – 1.66 (m, 4H). LC / MS: m / z=625.27[M+H] + .

[0201] Example 27: Preparation N -(2-aminophenyl)-4-[({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl]- H -pyrrolo[2,3- d ]pyrimidin-4-yl)piperidin-4-yl]carbonyl}amino)methyl]benzamide (Compound 27)

[0202]

[0203] Referring to the method of Example 1, intermediates 26-3 and 3-1 were used as raw materials to obtain compound 27 with a yield of 31.45%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.65 (brs, 1H), 9.62 (s, 1H), 8.45 (t, J = 6.0 Hz,1H), 8.33 (s, 1H), 7.92 (d, J= 7.9 Hz, 2H), 7.63 – 7.59 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.35 (d, J = 8.0 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21– 7.18 (m, 2H), 7.17 (d, J = 2.4 Hz, 1H), 7.16 – 7.13 (m, 1H), 7.03 (dd, J =8.4, 2.4 Hz, 1H), 6.99 – 6.95 (m, 1H), 6.79 – 6.77 (m, 1H), 6.62 – 6.58 (m,1H), 4.90 (s, 2H), 4.33 (d, J = 5.7 Hz, 2H), 4.28 (d, J = 13.0 Hz, 2H), 3.35– 3.34 (m, 1H), 3.04 – 2.90 (m, 2H), 1.76 – 1.71 (m, 4H).LC / MS: m / z=700.32[M+H] + .

[0204] Example 28: Preparation N -(2-amino-4-fluorophenyl)-4-[({[1-(5-{[2-chloro-4-(phenyloxy)phenyl]carbonyl}-7-yl]- H -pyrrolo[2,3- d ]pyrimidin-4-yl)piperidin-4-yl]carbonyl}amino)methyl]benzamide (Compound 28)

[0205]

[0206] Referring to the method of Example 1, intermediates 26-3 and 4-1 were used as raw materials to prepare compound 28 with a yield of 35.00%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.65 (brs, 1H), 9.55 (s, 1H), 8.45 (t, J = 6.1 Hz,1H), 8.32 (s, 1H), 7.92 (d, J = 8.1 Hz, 2H), 7.62 – 7.60 (m, 2H), 7.48 (dd,J = 8.6, 7.3 Hz, 2H), 7.34 (d, J = 7.9 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21– 7.18 (m, 2H), 7.17 (d, J = 2.4 Hz, 1H), 7.11 (dd, J = 8.7, 6.3 Hz, 1H),7.03 (dd, J = 8.4, 2.4 Hz, 1H), 6.54 (dd, J = 11.3, 2.9 Hz, 1H), 6.38 – 6.33(m, 1H), 5.23 (s, 2H), 4.33 (d, J = 5.7 Hz, 2H), 4.28 (d, J = 13.3 Hz, 2H),3.03 – 2.91 (m, 2H), 2.48 – 2.44 (m, 1H), 1.79 – 1.69 (m, 4H).LC / MS: m / z=718.36[M+H] + .

[0207] Example 29: Preparation (1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)- N -(4-(Hydroxyamino)-4-oxobutyl)cyclohexane-1-carboxamide (Compound 29)

[0208]

[0209] Step 1: Synthesis of intermediate 29-2

[0210] Referring to the method of Example 1, intermediate 1-5 and compound 29-1 were used as raw materials to prepare intermediate 29-2 with a yield of 99.70%. 1 H NMR (300 MHz, DMSO- d 6) δ 8.71 (d, J = 7.5 Hz, 1H), 8.23 ​​(s, 1H), 7.58 (d, J = 8.4 Hz, 2H), 7.48 (dd, J= 8.5, 7.3 Hz, 2H), 7.25 (t, J = 7.4Hz, 1H), 7.22 – 7.16 (m, 3H), 7.01 (dd, J = 8.4, 2.4 Hz, 1H), 4.03 – 3.96 (m,1H), 2.31 – 2.22 (m, 1H), 2.14 (dd, J = 12.2, 3.8 Hz, 2H), 1.98 (dd, J =13.3, 3.4 Hz, 2H), 1.55 – 1.42 (m, 2H), 1.41 – 1.27 (m, 2H).LC / MS: m / z=491.23[M+H] + .

[0211] Step 2: Synthesis of intermediate 29-3

[0212] Referring to the method of Example 1, intermediate 29-2 and 14-1 were used as raw materials to obtain intermediate 29-3 with a yield of 87.36%. LC / MS: m / z=590.34[M+H] + .

[0213] Step 3: Synthesis of Intermediate 29-4

[0214] Referring to the method of Example 14, intermediate 29-3 was used as a raw material to obtain intermediate 29-4 with a yield of 90.00%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.69 (d, J = 7.4 Hz, 1H), 8.23 ​​(s, 1H), 7.85 (t, J =5.5 Hz, 1H), 7.59 – 7.56 (m, 2H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.25 (t, J = 7.4 Hz, 1H), 7.21 – 7.16 (m, 3H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 4.00 –3.96 (m, 1H), 3.05 (q, J = 6.5 Hz, 2H), 2.19 – 2.11 (m, 5H), 1.82 (d, J=10.4 Hz, 2H), 1.65 – 1.57 (m, 2H), 1.57 – 1.47 (m, 2H), 1.34 – 1.24 (m, 2H).LC / MS: m / z=576.39[M+H] + .

[0215] Step 4: Synthesis of Intermediate 29-5

[0216] Referring to the method of Example 1, intermediate 29-4 and 1-8 were used as raw materials to obtain intermediate 29-5, with a yield of 59.91%. LC / MS: m / z=675.56[M+H] + .

[0217] Step 5: Synthesis of compound 29

[0218] Referring to the method of Example 1, intermediate 29-5 was used as raw material to prepare compound 29 with a yield of 81.77%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 10.39 (s, 1H), 8.73 (s, 1H), 8.69 (d, J = 7.4 Hz, 1H), 8.24 (s, 1H), 7.81 (t, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H),7.20 – 7.19 (m, 2H), 7.18 – 7.16 (m, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H),4.04 – 3.94 (m, 1H), 3.06 – 3.00 (m, 2H), 2.20 – 2.12 (m, 3H), 1.95 (t, J =7.5 Hz, 2H), 1.85 – 1.78 (m, 2H), 1.66 – 1.59 (m, 2H), 1.55 – 1.47 (m, 2H),1.39 – 1.22 (m, 2H).LC / MS: m / z=591.62[M+H] + .

[0219] Example 30: Preparation (1 R , 4 R )- N -(4-((2-aminophenyl)amino)-4-oxobutyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl) H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 30)

[0220]

[0221] Referring to the method of Example 1, intermediates 29-4 and 3-1 were used as raw materials to prepare compound 30 with a yield of 47.30%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.10 (s, 1H), 8.70 (d, J = 7.4 Hz,1H), 8.25 (s, 1H), 7.84 (t, J = 5.6 Hz, 1H), 7.62 (s, 1H), 7.58 (d, J = 8.4Hz, 1H), 7.49 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 –7.17 (m, 3H), 7.15 (dd, J = 7.9, 1.5 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H),6.92 – 6.87 (m, 1H), 6.73 – 6.70 (m, 1H), 6.56 – 6.51 (m, 1H), 4.87 (s, 2H),4.02 – 4.97(m, 1H), 3.11 (q, J = 6.6 Hz, 2H), 2.32 (t, J = 7.4 Hz, 2H), 2.25– 2.11 (m, 3H), 1.87 – 1.83 (m, 2H), 1.77 – 1.68 (m, 2H), 1.61 – 1.49 (m,2H), 1.41 – 1.22 (m, 2H).LC / MS: m / z=666.60[M+H] + .

[0222] Example 31: Preparation (1 R , 4 R )- N -(4-(2-amino-4-fluorophenyl)amino)-4-oxobutyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl) H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 31)

[0223]

[0224] Referring to the method of Example 1, intermediates 29-4 and 4-1 were used as raw materials to prepare compound 31 with a yield of 59.82%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.02 (s, 1H), 8.70 (d, J = 7.4 Hz,1H), 8.25 (s, 1H), 7.84 (t, J = 5.7 Hz, 1H), 7.62 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 –7.19 (m, 2H), 7.18 – 7.16 (m, 1H), 7.08 (dd, J = 8.7, 6.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.50 – 6.46 (m, 1H), 6.33 – 6.26 (m, 1H), 5.21 (s, 2H), 4.02 – 3.95 (m, 1H), 3.11 (q, J = 6.5 Hz, 2H), 2.30 (t, J = 7.4 Hz, 2H), 2.21– 2.08 (m, 3H), 1.87 – 1.82 (m, 2H), 1.76 – 1.67 (m, 2H), 1.61 – 1.49 (m,2H), 1.40 – 1.22 (m, 2H).LC / MS: m / z=684.58[M+H]+ .

[0225] Example 32: Preparation (1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)- N -(5-(Hydroxyamino)-5-oxopentyl)cyclohexane-1-carboxamide (Compound 32)

[0226]

[0227] Step 1: Synthesis of Intermediate 32-1

[0228] Referring to the method of Example 1, intermediates 29-2 and 17-1 were used as raw materials to obtain compound 32-1 with a yield of 73.14%. LC / MS: m / z=604.41[M+H] + .

[0229] Step 2: Synthesis of compound 32-2

[0230] Referring to the method of Example 14, intermediate 32-1 was used as a raw material to obtain intermediate 32-2 with a yield of 99.31%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.69 (d, J = 7.4 Hz, 1H), 8.23 ​​(s, 1H), 7.77 (t, J =5.6 Hz, 1H), 7.57 (d, J = 8.3 Hz, 2H), 7.51 – 7.45 (m, 2H), 7.26 (t, J = 7.4Hz, 1H), 7.21 – 7.16 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 4.01 – 3.96 (m,1H), 3.04 (q, J = 6.4 Hz, 2H), 2.21 – 2.12 (m, 5H), 1.84 – 1.79 (m, 2H), 1.57 – 1.45 (m, 4H), 1.43 – 1.38 (m, 2H), 1.34 – 1.25 (m, 2H). LC / MS: m / z=590.47[M+H] + .

[0231] Step 3: Synthesis of Intermediate 32-3

[0232] Referring to the method of Example 1, intermediates 32-2 and 1-8 were used as raw materials to obtain compound 32-3 with a yield of 49.52%. LC / MS: m / z=689.44[M+H] + .

[0233] Step 4: Synthesis of compound 32

[0234] Referring to the method of Example 1, compound 32 was prepared using intermediate 32-3 as raw material. The yield was 63.91%. 1 H NMR (300 MHz, DMSO- d 6 ) δ 12.73 (brs, 1H), 10.36 (s, 1H), 8.69 (d, J = 7.2 Hz,2H), 8.24 (s, 1H), 7.77 (t, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.4 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 –7.17 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 4.01 – 3.96 (m, 1H), 3.06 –2.99 (m, 2H), 2.22 – 2.11 (m, 3H), 1.95 (t, J = 7.1 Hz, 2H), 1.83 – 1.80 (m,2H), 1.57 – 1.28 (m, 8H).LC / MS: m / z=605.37[M+H] + .

[0235] Example 33: Preparation (1 R , 4 R )- N -(5-(2-aminophenyl)-5-oxopentyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 33)

[0236]

[0237] Referring to the method of Example 1, intermediates 32-2 and 3-1 were used as raw materials to prepare compound 33 with a yield of 65.32%. 1 HNMR (400 MHz, DMSO- d 6) δ 12.71 (brs, 1H), 9.09 (s, 1H), 8.69 (d, J = 7.4 Hz,1H), 8.25 (s, 1H), 7.79 (t, J = 5.6 Hz, 1H), 7.60 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.4 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 –7.15 (m, 4H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.91 – 6.87 (m, 1H), 6.73 –6.71 (m, 1H), 6.56 – 6.52 (m, 1H), 4.83 (s, 2H), 4.03 – 3.96 (m, 1H), 3.08(q, J = 6.6 Hz, 2H), 2.33 (t, J = 7.3 Hz, 2H), 2.23 – 2.12 (m, 3H), 1.88 –1.79 (m, 2H), 1.62 – 1.42 (m, 6H), 1.31 (dt, J = 12.9, 9.6 Hz, 2H).LC / MS: m / z=680.42[M+H] + .

[0238] Example 34: Preparation (1 R , 4 R )- N -(5-(2-aminophenyl)-5-oxopentyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 34)

[0239]

[0240] Referring to the method of Example 1, intermediates 32-2 and 4-1 were used as raw materials to prepare compound 33 with a yield of 47.04%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.04 (s, 1H), 8.69 (d, J = 7.4 Hz,1H), 8.24 (s, 1H), 7.84 – 7.78 (m, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5 Hz,1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.16(m, 3H), 7.10 (dd, J = 8.7, 6.4 Hz, 1H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 6.51– 6.46 (m, 1H), 6.34 – 6.27 (m, 1H), 5.16 (s, 2H), 4.08 – 3.92 (m, 1H), 3.11– 3.04 (m, 2H), 2.31 (t, J = 7.3 Hz, 2H), 2.24 – 2.11 (m, 3H), 1.84 – 1.81(m, 2H), 1.63 – 1.41 (m, 6H), 1.36 – 1.25 (m, 2H).LC / MS: m / z=698.40[M+H] + .

[0241] Example 35: Preparation (1 R , 4 R )- N -(5-(2-amino-4-fluorophenyl)amino)-5-oxopentyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl) H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 35)

[0242]

[0243] Step 1: Synthesis of Intermediate 35-1

[0244] Referring to the method of Example 1, intermediate 29-2 and 20-1 were used as raw materials to obtain intermediate 35-1 with a yield of 79.42%. LC / MS: m / z=618.42[M+H] + .

[0245] Step 2: Synthesis of Intermediate 35-2

[0246] Referring to the method of Example 14, intermediate 35-1 was used as a raw material to obtain intermediate 35-2 with a yield of 94.21%. 1 HNMR (400 MHz, DMSO- d 6) δ 8.69 (d, J = 7.4 Hz, 1H), 8.24 (s, 1H), 7.73 (t, J =5.6 Hz, 1H), 7.61 – 7.56 (m, 2H), 7.48 (dd, J = 8.5, 7.4 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.21 – 7.17 (m, 3H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 4.00 –3.96 (m, 1H), 3.03 (q, J = 6.5 Hz, 2H), 2.20 – 2.11 (m, 5H), 1.84 – 1.79 (m,2H), 1.57 – 1.45 (m, 4H), 1.39 (p, J = 7.1 Hz, 2H), 1.35 – 1.22 (m, 4H). LC / MS: m / z=604.48[M+H] + .

[0247] Step 3: Synthesis of Intermediate 35-3

[0248] Referring to the method of Example 1, intermediate 35-2 and 1-8 were used as raw materials to obtain intermediate 35-3, with a yield of 63.44%. LC / MS: m / z=703.52[M+H] + .

[0249] Step 4: Synthesis of compound 35

[0250] Referring to the method of Example 1, compound 35 was prepared using intermediate 35-3 as raw material. The yield was 87.67%. 1 H NMR (300 MHz, DMSO- d6) δ 12.75 (s, 1H), 10.36 (s, 1H), 8.71 (d, J = 7.4 Hz, 2H),8.25 (s, 1H), 7.75 (t, J = 5.6 Hz, 1H), 7.62 (s, 1H), 7.58 (d, J = 8.5 Hz,1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 – 7.16(m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 4.01 – 3.94 (m, 1H), 3.02 (q, J =6.5 Hz, 2H), 2.20 – 2.11 (m, 3H), 1.94 (t, J = 7.3 Hz, 2H), 1.88 – 1.75 (m,2H), 1.60 – 1.19 (m, 10H).LC / MS: m / z=619.64[M+H] + .

[0251] Example 36: Preparation (1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)- N -(6-(Hydroxyamino)-6-oxohexyl)cyclohexane-1-carboxamide (Compound 36)

[0252]

[0253] Referring to the method of Example 1, intermediates 35-2 and 3-1 were used as raw materials to prepare compound 36 with a yield of 22.71%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.72 (brs, 1H), 9.10 (s, 1H), 8.69 (d, J = 7.4 Hz,1H), 8.25 (s, 1H), 7.77 (t, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J= 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 –7.14 (m, 4H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 6.91 – 6.86 (m, 1H), 6.73 –6.70 (m, 1H), 6.56 – 6.51 (m, 1H), 4.83 (s, 2H), 4.04 – 3.94 (m, 1H), 3.09 –3.02 (m, 2H), 2.31 (t, J = 7.4 Hz, 2H), 2.19 – 2.12 (m, 3H), 1.88 – 1.77 (m,2H), 1.64 – 1.25 (m, 10H).LC / MS: m / z=694.50[M+H] + .

[0254] Example 37: Preparation (1 R , 4 R )- N -(6-(2-amino-4-fluorophenyl)amino)-6-oxohexyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7-yl)- H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 37)

[0255]

[0256] Referring to the method of Example 1, intermediates 35-2 and 4-1 were used as raw materials to prepare compound 37 with a yield of 19.51%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.03 (s, 1H), 8.69 (d, J = 7.3 Hz,1H), 8.24 (s, 1H), 7.77 (t, J = 5.7 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J= 7.4 Hz, 1H), 7.22 –7.16 (m, 3H), 7.10 (dd, J = 8.7, 6.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H),6.51 – 6.46 (m, 1H), 6.33 – 6.27 (m, 1H), 5.14 (s, 2H), 4.01 – 3.91 (m, 1H),3.05 (q, J = 6.4 Hz, 2H), 2.29 (t, J = 7.4 Hz, 2H), 2.23 – 2.10 (m, 3H), 1.85 – 1.80 (m, 2H), 1.65 – 1.39 (m, 6H), 1.36 – 1.23 (m, 4H). LC / MS: m / z=712.47[M+H] + .

[0257] Example 38: Preparation (1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)- N -(7-(Hydroxyamino)-7-oxoheptyl)cyclohexane-1-carboxamide (Compound 38)

[0258]

[0259] Step 1: Synthesis of Intermediate 38-1

[0260] Referring to the method of Example 1, intermediates 29-2 and 23-1 were used as raw materials to obtain compound 38-1 with a yield of 77.66%. LC / MS: m / z=632.50[M+H] + .

[0261] Step 2: Synthesis of Intermediate 38-2

[0262] Referring to the method of Example 14, compound 38-2 was prepared using intermediate 38-1 as raw material with a yield of 96.88%. 1 HNMR (300 MHz, DMSO- d 6) δ 8.69 (d, J = 7.4 Hz, 1H), 8.22 (s, 1H), 7.76 (t, J=5.6 Hz, 1H), 7.60 – 7.55 (m, 2H), 7.51 – 7.45 (m, 2H), 7.25 (t, J = 7.4 Hz,1H), 7.21 – 7.16 (m, 3H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 4.00 – 3.95 (m,1H), 3.02 (q, J = 6.0, 5.6 Hz, 2H), 2.19 – 2.11 (m, 5H), 1.83 – 1.78 (m, 2H), 1.60 – 1.45 (m, 4H), 1.45 – 1.29 (m, 4H), 1.27 – 1.22 (m, 4H).LC / MS: m / z=618.42[M+H] + .

[0263] Step 3: Synthesis of Intermediate 38-3

[0264] Referring to the method of Example 1, intermediates 38-2 and 1-8 were used as raw materials to obtain compound 38-3, with a yield of 66.62%. LC / MS: m / z=717.72[M+H] + .

[0265] Step 4: Synthesis of compound 38

[0266] Referring to the method of Example 1, compound 38 was prepared using intermediate 38-3 as raw material. The yield was 76.94%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.75 (brs, 1H), 10.35 (s, 1H), 8.72 (d, J = 7.6 Hz,1H), 8.25 (s, 1H), 7.75 (t, J = 5.6 Hz, 1H), 7.62 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 –7.15 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H), 4.03 – 3.93 (m, 1H), 3.02 (q,J = 6.6 Hz, 2H), 2.23 – 2.11 (m, 3H), 1.94 (t, J = 7.3 Hz, 2H), 1.87 – 1.76 (m,2H), 1.60 – 1.42 (m, 4H), 1.42 – 1.20 (m, 8H).LC / MS: m / z=633.52[M+H] + .

[0267] Example 39: Preparation (1 R , 4 R )- N -(7-(2-aminophenyl)-7-oxoheptyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 39)

[0268]

[0269] Referring to the method of Example 1, intermediates 38-2 and 3-1 were used as raw materials to prepare compound 39 with a yield of 68.42%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.11 (s, 1H), 8.69 (d, J = 7.4 Hz,1H), 8.24 (s, 1H), 7.76 (t, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22 –7.13 (m, 4H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 6.92 – 6.86 (m, 1H), 6.73 –6.70 (m, 1H), 6.56 – 6.51 (m, 1H), 4.83 (s, 2H), 4.06 – 3.92 (m, 1H), 3.04(q, J = 6.4 Hz, 2H), 2.31 (t, J= 7.4 Hz, 2H), 2.24 – 2.10 (m, 3H), 1.84 –1.80 (m, 2H), 1.64 – 1.47 (m, 4H), 1.46 – 1.24 (m, 8H). LC / MS: m / z=708.70[M+H] + .

[0270] Example 40: Preparation (1 R , 4 R )- N -(7-(2-amino-4-fluorophenyl)amino)-7-oxoheptyl)-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide (Compound 40)

[0271]

[0272] Referring to the method of Example 1, intermediates 38-2 and 4-1 were used as raw materials to prepare compound 40 with a yield of 69.83%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.03 (s, 1H), 8.69 (d, J = 7.4 Hz,1H), 8.24 (s, 1H), 7.76 (t, J = 5.6 Hz, 1H), 7.61 (s, 1H), 7.58 (d, J = 8.5Hz, 1H), 7.48 (dd, J = 8.5, 7.3 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.19 (dt, J = 5.7, 1.3 Hz, 3H), 7.09 (dd, J = 8.7, 6.3 Hz, 1H), 7.02 (dd, J = 8.5, 2.4Hz, 1H), 6.51 – 6.46 (m, 1H), 6.34 – 6.27 (m, 1H), 5.14 (s, 2H), 4.01 – 3.94(m, 1H), 3.04 (q, J = 6.6 Hz, 2H), 2.29 (t, J= 7.4 Hz, 2H), 2.23 – 2.08 (m,3H), 1.84 – 1.80 (m, 2H), 1.64 – 1.48 (m, 4H), 1.44 – 1.23 (m, 8H).LC / MS: m / z=726.68[M+H] + .

[0273] Example 41: Preparation of 4-((1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide)- N -Hydroxybenzamide (Compound 41)

[0274]

[0275] Step 1: Synthesis of Intermediate 41-1

[0276] Referring to the method of Example 1, intermediate 41-1 was prepared using intermediates 29-2 and 26-1 as raw materials, with a yield of 53.86%. LC / MS: m / z=638.51[M+H] + .

[0277] Step 2: Synthesis of Intermediate 41-2

[0278] Referring to the method of Example 14, intermediate 41-1 was used as a raw material to obtain intermediate 41-2 with a yield of 73.47%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.79 (s, 1H), 8.71 (d, J = 7.4 Hz, 1H), 8.43 (t, J = 6.0 Hz, 1H), 8.25 (s, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.65 – 7.56 (m, 2H), 7.53 – 7.44 (m, 2H), 7.34 (d, J = 8.1 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.22– 7.16 (m, 3H), 7.02 (dd, J = 8.4, 2.4 Hz, 1H), 4.34 (d,J = 5.7 Hz, 2H),4.04 – 3.99 (m, 1H), 2.34 – 2.25 (m, 1H), 2.24 – 2.14 (m, 2H), 1.96 – 1.84(m, 2H), 1.67 – 1.50 (m, 2H), 1.42 – 1.25 (m, 2H).

[0279] Step 3: Synthesis of Intermediate 41-3

[0280] Referring to the method of Example 1, intermediate 41-2 and 1-8 were used as raw materials to obtain intermediate 41-3, with a yield of 56.40%. LC / MS: m / z=723.54[M+H] + .

[0281] Step 4: Synthesis of compound 41

[0282] Referring to the method of Example 1, intermediate 41-3 was used as raw material to prepare compound 41 with a yield of 69.14%. 1 H NMR (300 MHz, DMSO- d 6) δ 12.74 (s, 1H), 11.19 (s, 1H), 9.02 (s, 1H), 8.71 (d, J =7.3 Hz, 1H), 8.40 (t, J = 6.0 Hz, 1H), 8.25 (s, 1H), 7.71 (d, J = 8.3 Hz, 2H), 7.62 (s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.49 (dd, J = 8.5, 7.3 Hz, 2H),7.34 – 7.23 (m, 3H), 7.22 – 7.16 (m, 3H), 7.02 (dd, J = 8.5, 2.4 Hz, 1H),4.31 (d, J = 5.6 Hz, 2H), 4.04 – 3.98 (m, 1H), 2.33 – 2.13 (m, 3H), 1.91 –1.87 (m, 2H), 1.42 – 1.25 (m, 2H). LC / MS: m / z=639.54[M+H] + .

[0283] Example 42: Preparation N-(2-aminophenyl)-4-((1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H -pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide)methyl)benzamide (Compound 42)

[0284]

[0285] Referring to the method of Example 1, intermediates 41-2 and 3-1 were used as raw materials to prepare compound 42 with a yield of 23.68%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.74 (brs, 1H), 9.64 (s, 1H), 8.71 (d, J = 7.3 Hz,1H), 8.45 (d, J = 5.4 Hz, 1H), 8.25 (s, 1H), 7.94 (d, J = 8.0 Hz, 2H), 7.62(s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.53 – 7.44 (m, 2H), 7.37 (d, J = 8.1 Hz,2H), 7.26 (t, J = 7.4 Hz, 1H), 7.23 – 7.15 (m, 4H), 7.07 – 6.93 (m, 2H), 6.80 – 6.77 (m, 1H), 6.65 – 6.56 (m, 1H), 4.91 (s, 2H), 4.35 (d, J = 5.2 Hz, 2H),4.09 – 3.93 (m, 1H), 2.36 – 2.15 (m, 3H), 1.93 – 1.88 (m, 2H), 1.65 – 1.53(m, 2H), 1.40 – 1.33 (m, 2H).LC / MS: m / z=714.52[M+H] + .

[0286] Example 43: Preparation N -(2-amino-4-fluorophenyl)-4-((1 R , 4 R )-4-(5-(2-chloro-4-phenoxybenzoyl)-7 H-pyrrolo[2,3- d ]pyrimidin-4-yl)amino)cyclohexane-1-carboxamide)methyl)benzamide (Compound 43)

[0287]

[0288] Referring to the method of Example 1, intermediates 41-2 and 4-1 were used as raw materials to prepare compound 43 with a yield of 19.35%. 1 HNMR (300 MHz, DMSO- d 6) δ 12.73 (brs, 1H), 9.57 (s, 1H), 8.71 (d, J = 7.4 Hz,1H), 8.44 (t, J = 6.0 Hz, 1H), 8.25 (s, 1H), 7.94 (d, J = 8.0 Hz, 2H), 7.62(s, 1H), 7.59 (d, J = 8.5 Hz, 1H), 7.49 (dd, J = 8.5, 7.3 Hz, 2H), 7.36 (d, J = 8.0 Hz, 2H), 7.26 (t, J = 7.4 Hz, 1H), 7.23 – 7.17 (m, 3H), 7.12 (dd, J =8.7, 6.3 Hz, 1H), 7.03 (dd, J = 8.5, 2.4 Hz, 1H), 6.57 – 6.52 (m, 1H), 6.40 –6.33 (m, 1H), 5.24 (s, 2H), 4.39 – 4.34 (m, 2H), 4.11 – 3.95 (m, 1H), 2.35 –2.15 (m, 3H), 1.92 – 1.88 (m, 2H), 1.65-1.52 (m, 2H), 1.40 – 1.29 (m, 2H).LC / MS: m / z=732.50[M+H] + .

[0289] 2. Biological Activity Test

[0290] 1. In vitro BTK kinase activity inhibition assay

[0291] 1: Preparation of compound storage solution

[0292] Dissolve the compound powder in 100% DMSO to prepare a 10 mM stock solution and store at -20℃ in the dark.

[0293] 2: Kinase reaction process

[0294] BTK used wt Kinases and BTK C481S The kinases were purchased from Promega, and the two types of BTK kinase reactions were performed according to the following steps:

[0295] (1) Prepare 4X kinase buffer and 1X kinase buffer; (2) Preparation of compound solution: The concentration of the tested compound is 50 μM. It is first diluted to 100 times the final concentration of 100% DMSO solution, and then diluted to 5 times the final concentration of 5% DMSO solution using 4X kinase buffer. Transfer 1 μL to a 384-well plate with a dispenser, with duplicate wells for each compound; (3) Prepare a kinase solution with a final concentration of 2.5 times using 4X kinase buffer; (4) Add 2 μL of the kinase solution with a final concentration of 2.5 times to the compound and positive control wells, and add 3 μL of 1X kinase buffer to the negative control wells; (5) Vortex the reaction plate to mix well and incubate at room temperature for 10 minutes; (6) Prepare a mixed solution of ATP and Kinase substrate with a final concentration of 2.5 times using 4X kinase buffer; (7) Add 2 μL of ATP and Kinase (8) Mix the solution of substrate to start the reaction; (9) Add 5 μL ADP-Glo ​​reagent, mix it by shaking, terminate the reaction, and incubate it at room temperature for 60 min to consume the residual ATP; (10) Add 10 μL kinase detection reagent, mix it by shaking, and incubate it at room temperature for 40 min; (11) Read the data using an ELISA reader and calculate the inhibition rate.

[0296] 3: Data Analysis

[0297] Calculation formula:

[0298] Wherein: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min: the mean of the negative control wells, representing the conversion rate reading of the wells without enzyme activity; Conversion%_max: the mean of the ratio of the negative control wells, representing the conversion rate reading of the wells without compound inhibition; The results of the inhibition of two types of kinase activity of BTK in vitro by the compounds prepared in Examples 1-43 are shown in Table 1 below.

[0299] (II) In vitro BTK kinase activity inhibition comparison test

[0300] The specific experimental method is basically the same as that of the inhibition test (I), except that the compound solution to be tested is replaced with a self-prepared ARQ531 inhibitor solution, and the preparation method provided in the Chinese patent publication with application number 2015800858320 is adopted. After comparative experiments, the inhibition results of the ARQ531 inhibitor on two types of BTK kinase activities in vitro are shown in Table 1 below.

[0301] (III) In vitro HDAC enzyme activity inhibition assay

[0302] 1: Compound solution preparation

[0303] The compounds prepared in Examples 1-43 were weighed to prepare 10 mM DMSO stock solutions, 1 μL was added to 999 μL of deionized water (diluted 1000 times) to obtain a 10 μM compound solution, and 10 μL of the solution was added to 500 μL of deionized water (diluted 50 times) to obtain a 200 nM compound solution (2 times the final concentration).

[0304] 2: Kinase experiment part

[0305] (1) Pipette 50 μL of compound and add it to a 96-well plate; (2) 2 μL HDAC Inhibitor (purchased from abcam) and 48 μL deionized water as a negative control group (N) were added to a 96-well plate; (3) 50 μL deionized water was added to a 96-well plate as a positive control group (P); (4) 33 μL deionized water, 2 μL HeLa Nuclear Extract, 10 μL 10X HDAC Assay Buffer and 5 μL HDAC Substrate were added to the 96-well plate and mixed immediately and added to the 96-well plate to mix with the compound; (5) Incubate at 37 °C for 30 min; (6) Add 10 μL Lysine Developer to each well and mix; (7) Incubate at 37 °C for 30 min; (8) Read the luminescence value RLU (Ex / Em = 360 / 450 nm) with an enzyme reader.

[0306] Calculation formula: %Inhibition=100-[RLU−Mean(NC)] / [Mean(PC)−Mean(NC)]×100

[0307] Wherein: RLU: chemiluminescence value of the sample; Mean (NC): mean value of the negative control wells; Mean (PC): mean value of the positive control wells; The results of the inhibition of HDAC enzyme activity in vitro by the compounds prepared in Examples 1-43 are shown in Table 1 below.

[0308] (IV) In vitro HDAC enzyme activity inhibition comparison test

[0309] The specific experimental method is basically the same as that of inhibition test (III), except that the solution of the compound to be tested is replaced with SAHA solution. SAHA was purchased from Anage Chemical Company. The results of the comparative test on the inhibition of HDAC enzyme activity in vitro by SAHA are shown in Table 1 below.

[0310] Table 1

[0311] Compound <![CDATA[BTK wt (10nM) inhibition rate]]> <![CDATA[BTK C481S (10 nM)]]> Inhibition rate of HDAC (50nM) Compound <![CDATA[BTK wt (10nM) inhibition rate]]> <![CDATA[BTK C481S (10 nM)]]> Inhibition rate of HDAC (50nM) Compound <![CDATA[BTK wt (10nM) inhibition rate]]> <![CDATA[BTK C481S (10 nM)]]> Inhibition rate of HDAC (50nM) 1 45.66% 44.26% 77.16% 16 2.45% 48.88% 6.56% 31 28.03% 53.69% 49.39% 2 39.54% 47.90% 96.98% 17 45.01% 46.61% 89.86% 32 31.20% 48.45% 84.12% 3 30.54% 38.18% 42.26% 18 4.91% 47.32% 29.69% 33 42.02% 42.00% 24.06% 4 30.03% 53.07% 11.27% 19 46.84% 37.16% <5% 34 33.30% 37.67% 27.06% 5 44.50% 63.61% 97.88% 20 3.60% 34.44% 88.55% 35 36.54% 40.62% 94.70% 6 35.60% 31.10% 34.50% 21 43.01% 47.41% 5.47% 36 54.58% 38.61% 35.16% 7 33.10% 55.57% 18.68% 22 13.32% 30.99% 106.09% 37 50.11% 53.14% 20.71% 8 44.47% 48.67% 96.21% 23 46.26% 52.40% 107.77% 38 57.63% 54.58% 93.99% 9 36.89% 46.46% 19.40% 24 32.46% 41.48% 48.98% 39 36.75% 32.05% 27.09% 10 44.26% 65.39% 9.96% 25 7.24% 49.55% 48.91% 40 43.30% 42.15% 22.57% 11 29.32% 39.03% 57.44% 26 0.15% 38.84% 106.71% 41 32.99% 42.95% 72.01% 12 50.50% 33.90% 3.78% 27 53.62% 50.12% 74.34% 42 42.54% 47.55% 47.08% 13 8.47% ＜5% ＜5% 28 32.79% 60.13% 66.72% 43 53.26% 60.12% 30.91% 14 1.16% 28.76% 76.06% 29 25.34% 47.38% 53.74% ARQ531 69.86% 67.43% ND 15 25.07% 47.19% 21.46% 30 34.09% 34.45% 19.28% SAHA ND ND 95.11%

[0312] As can be seen from the data in Table 1, ARQ531 is a clinical drug targeting BTK, which is used as a reference for the BTK inhibition rate of the compound. SAHA is a marketed drug targeting HDAC, which is used as a reference for the inhibition rate of HDAC of the compound. The single compound of this scheme has a dual inhibitory effect on BTK and HDAC, and the inhibitory effect of a single kinase of some compounds is comparable to or even better than that of positive drugs. The successful design of dual-target compounds has been demonstrated, and the inhibitory effects of single compounds such as 17, 23, 35, 38, etc. on both kinases are close to those of positive drugs.

[0313] (V) Determination of the inhibitory activity of compounds on in vitro cell proliferation (SU-DHL-4)

[0314] 1: SU-DHL-4 diffuse large B-cell lymphoma cell line was purchased from Guangzhou Geneo Biotechnology Co., Ltd. and cultured in RPMI-1640 plus 10% fetal bovine serum (Gibcol) at 37°C, 5% CO2, and 95% humidity.

[0315] Cell lines Cell Type Number of cells / well Culture medium SU-DHL-4 Suspension 10000 RPMI-1640 + 10% FBS

[0316] 2: The following are general experimental steps

[0317] The cells in the logarithmic growth phase were harvested and the cell viability was detected by trypan blue staining. The cells were counted using a cell counter (thermo countness II). The appropriate amount of cells was taken and the cell concentration was adjusted. 50 μL of the cell suspension was added to a 96-well plate, 5000 cells per well, and cultured overnight.

[0318] The compounds prepared in Example 1-43 were formulated into test compound solutions, and 50 μL of the test compound solutions were added to each well of cells. The final concentration of each test compound was 0.5 μM, and three replicate wells were set for each compound concentration. The cells were cultured in an incubator for 72 hours.

[0319] After 72 hours of culture after administration, 10 μL of CCK-8 solution was added to each well, and the 96-well plate was placed in an incubator. After 2 hours of continued culture, the absorbance was read at 450 nm and the relative inhibition rate was calculated; the results are shown in Table 2 below.

[0320] 6. Comparative experimental determination of in vitro cell proliferation (SU-DHL-4) inhibitory activity

[0321] The cells and experimental steps used were basically the same as those of the assay experiment (V), except that the compound solutions used were replaced with ARQ531 solution and SAHA solution, respectively, with a final concentration of 0.5 μM. The assay results are shown in Table 2.

[0322] Table 2

[0323] Compound The cell activity inhibition rate at a concentration of 0.5 μM <![CDATA[IC 50 (nM)]]> Compound The cell activity inhibition rate at a concentration of 0.5 μM <![CDATA[IC 50 (nM)]]> Compound The cell activity inhibition rate at a concentration of 0.5 μM <![CDATA[IC 50 (nM)]]> Compound The cell activity inhibition rate at a concentration of 0.5 μM <![CDATA[IC 50 (nM)]]> 1 60.32% 367.9 13 8.61% NT 25 19.59% 982.5 37 71.40% 68.99 2 46.22% 277.7 14 <5% NT 26 <5% NT 38 37.72% 76.7 a3 19.55% 1336 15 33.05% 4244 27 53.46% 807 39 21.49% 106.6 4 12.01% 1276 16 35.54% 2305 28 19.85% 1189 40 51.77% 221.6 5 67.14% 1498 17 <5% NT 29 23.81% NT 41 8.93% 1889 6 17.98% 1639 18 9.77% NT 30 37.91% 949.8 42 42.86% 109.7 7 10.69% NT 19 13.34% NT 31 35.54% 928.9 43 11.78% 136.7 8 19.35% 33283 20 20.12% NT 32 6.55% NT ARQ531 71.29% 116 9 26.87% 1079 21 12.75% 1047 33 41.71% 900.5 SAHA NT 688.4 10 11.96% 1509 22 13.89% NT 34 <5% 904.3 11 16.73% 1049 23 5.53% 3059 35 19.30% 571.4 12 14.52% 2932 24 25.68% 694.2 36 70.31% 97.22

[0324] From the data in Table 2, we can see that B cell malignancies were selected as indications, and human B lymphoma SU-DHL-4 cells were selected as test cells to verify the tumor inhibition effect of the compound. It can be seen that such compounds have a certain killing effect on tumor cells, and single compounds such as 37, 38, 39, and 42 have better inhibition on tumor cells than two positive drugs.

[0325] It can be seen from the above examples that the compounds prepared in Examples 1-43 of the present invention have strong inhibitory effects on both BTK kinase and HDAC enzyme, can be used as BTK / HDAC dual inhibitors, and have considerable anti-tumor cell proliferation activity, and can be used to prepare drugs for treating diseases caused by overexpression of BTK kinase and / or HDAC enzyme.

Claims

1. A (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound or its tautomer or pharmaceutically acceptable salt, characterized in that: The structural formula of the (4-phenoxyphenyl)pyrrolopyrimidin-5-yl)methanone compound is as shown in Formula I: Wherein: at least one of W and Y exists, and W is selected from the following linking groups: The Y is selected from the following linking groups: The N atom in the structural fragment defined by W is connected to pyrrolopyrimidine, the left side of the structural fragment defined by Y is connected to the RNH acyl group and the right side is connected to the W structure; each n is independently selected from 1-6; The R is selected from hydroxyl, 2-aminophenyl, 2-amino-4-fluorophenyl; The R a , R b are independently selected from: hydrogen, halogen, cyano, nitro, hydroxyl, carboxyl, substituted or unsubstituted C 1-6 acyl, substituted or unsubstituted amino, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 one or more of the alkoxy groups; The substituted C 1-6 Acyl, substituted amino, substituted C 1-6 Alkyl, substituted C 1-6 The substituents in the alkoxy group are selected from one or more of the following groups: 1-5 Alkyl, C 2-5 Alkenyl, C 2-5 Alkynyl, C 1-5 Alkoxy, halogen, nitro, cyano, hydroxy, amino, carboxyl, carbonyl, ester, amide and oxo.

2. The (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound or its tautomer or pharmaceutically acceptable salt according to claim 1, characterized in that: At least one of W and Y exists, and W is selected from the following linking groups: The Y is selected from the following linking groups: Each n is independently selected from 1-6; The R is selected from hydroxyl, 2-aminophenyl, 2-amino-4-fluorophenyl; The R a , R b Independently selected from: one or more of hydrogen, halogen, cyano, nitro, hydroxyl, and carboxyl; The halogen is selected from F, Cl, and Br.

3. The (4-phenoxyphenyl)(pyrrolopyrimidin-5-yl)methanone compound or its tautomer or pharmaceutically acceptable salt according to claim 1, characterized in that: The compound is selected from 1 to 43:

4. A method for preparing the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound having the structure of formula I as claimed in claim 1 or its tautomer, pharmaceutically acceptable salt, comprising the following steps: First, compound A is used as a raw material, and a halogen-metal exchange reaction is carried out with n-butyl lithium to form an organic lithium compound, and the organic lithium compound then undergoes a nucleophilic addition reaction with 4-phenoxybenzoate to obtain compound B; then, a weak base is used as an acid-binding agent, and compound B undergoes a nucleophilic substitution reaction with a compound containing an amine structure to obtain a compound of formula I.

5. A combination drug, characterized in that: The active ingredient is selected from the (4-phenoxyphenyl) (pyrrolopyrimidin-5-yl) ketone compound or its tautomer, and a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3.

6. Use of the compound according to any one of claims 1 to 3 or its tautomers or pharmaceutically acceptable salts in the preparation of a medicament for treating a disease caused by overexpression of BTK kinase and / or HDAC enzyme, or use of the combination drug according to claim 5 in the preparation of a medicament for treating a disease caused by overexpression of BTK kinase and / or HDAC enzyme; the disease is selected from one or more of autoimmune diseases, inflammatory diseases, thromboembolic diseases, allergies, infectious diseases, proliferative diseases and cancer.

7. The use according to claim 6, characterized in that: The disease is selected from arthritis, rheumatoid arthritis, urticaria, vitiligo, organ transplant rejection, ulcerative colitis, Crohn's disease, dermatitis, asthma, Sjögren's syndrome, systemic lupus erythematosus, multiple sclerosis, idiopathic thrombocytopenic purpura, rash, anti-neutrophil cytoplasmic antibody vasculitis, pus, pus, chronic obstructive pulmonary disease, psoriasis; breast cancer, mantle cell lymphoma, ovarian cancer, esophageal cancer, laryngeal cancer, glioblastoma, neuroblastoma, gastric cancer, hepatocellular carcinoma, glioma, endometrial cancer, melanoma, kidney cancer, bladder cancer, biliary tract cancer. Cancer, pancreatic cancer, lymphoma, hairy cell cancer, nasopharyngeal cancer, colorectal cancer, rectal cancer, brain and central nervous system cancer, cervical cancer, prostate cancer, testicular cancer, genitourinary tract cancer, lung cancer, small cell lung cancer, small cell carcinoma, lung adenocarcinoma, bone cancer, colon cancer, adenoma, pancreatic cancer, thyroid cancer, follicular carcinoma, Hodgkin's leukemia, bronchial cancer, uterine corpus cancer, cervical cancer, multiple myeloma, acute myeloid leukemia, chronic myeloid leukemia, lymphocytic leukemia, chronic lymphoid leukemia, myeloid leukemia, non-Hodgkin's lymphoma, primary macroglobulinemia.

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