Difluoromethoxy phenyl PDE3 and / or PDE4 inhibitor and application thereof

Abstract

The invention discloses a difluoromethoxy phenyl PDE3 and / or PDE4 inhibitor and application thereof. The difluoromethoxy phenyl PDE3 and / or PDE4 inhibitor can prevent and / or treat inflammatory diseases, respiratory diseases, skin diseases or immune system diseases.

Description

Technical Field

[0001] The present application belongs to the field of medical technology, and specifically relates to difluoromethoxyphenyl derivatives, which are inhibitors of PDE3 and / or PDE4. Background Art

[0002] Phosphodiesterase (PDE) is a type of hydrolase that hydrolyzes cAMP and cGMP. It plays a key role in intracellular signal transduction by selectively hydrolyzing the second messengers cAMP and cGMP that regulate proteins and transcription factors. Since EW Sutherland proposed the concept of second messenger based on the important role of cyclic adenosine monophosphate (cAMP) in cells in 1965, "second messenger" has become an extremely important tool for drug discovery. The second messengers mainly include: cyclic adenosine monophosphate, cyclic guanosine monophosphate (cGMP), inositol triphosphate (IP3), diacylglycerol (DAG), calcium ions (Ca 2+ ). Second messengers respond to the binding of extracellular signals to cell surface receptors through changes in their concentration (increase or decrease), regulate the activity of intracellular enzymes and non-enzyme proteins, and thus perform the function of carrying and amplifying signals in the cell signal transduction pathway. Phosphodiesterase has the function of hydrolyzing intracellular cAMP and cGMP, degrading intracellular cAMP or cGMP into 5-monophosphate nucleoside (5'AMP or 5'GMP), thereby terminating the biochemical effects transmitted by these second messengers. cAMP-PDE and cGMP-PDE have long been considered targets for drug development.

[0003] PDE3 is an enzyme that can hydrolyze both cAMP and cGMP. However, it hydrolyzes cAMP 10 times faster than cGMP. There are two subtypes of PDE3: PDE3A (mainly found in smooth muscle) and PDE3B (mainly found in adipose tissue). PDE3 is mainly concentrated in alveolar macrophages, endothelial cells, and platelets in the respiratory system; it is also involved in regulating cardiac contractility and vascular smooth muscle. Selective PDE3 inhibitors include amrinone, milrinone, cilostazol, and cilostazol. A large number of studies have found that PDE3 isoenzymes are considered to be dominant in the function of airway smooth muscle, and PDE3 inhibitors can relax airway smooth muscle and are now being developed as bronchodilators.

[0004] PDE4 is an enzyme that specifically hydrolyzes cAMP. Its family consists of four subtypes: PDE4A, PDE4B, PDE4C, and PDE4D. Each subtype has a corresponding gene encoding and has different cell distribution and functions. PDE4 is mainly distributed in airway smooth muscle cells and inflammatory cells and immune cells such as lymphocytes, mast cells, macrophages, neutrophils, eosinophils, basophils, monocytes, and epithelial cells, regulating the level of cAMP in these cells. Inhibition of PDE4 can cause an increase in the level of cAMP in inflammatory cells and immune regulatory cells, thereby inhibiting the function of inflammatory cells and relaxing airway smooth muscle. PDE4 is also expressed in fibroblasts. Selective PDE4 inhibitors can effectively inhibit the production of matrix metalloproteinases, showing the therapeutic value of inhibiting tissue remodeling in related lung diseases.

[0005] Bronchial asthma, referred to as asthma, is a heterogeneous disease with respiratory symptoms such as cough, chest tightness, wheezing, and shortness of breath as the main clinical manifestations. It recurs repeatedly and is difficult to cure. Asthma is prevalent worldwide. There are about 310 million bronchial asthma patients worldwide, and about 30 million people in my country suffer from bronchial asthma. Bronchial asthma is an allergic disease of the lungs and respiratory tract. The main characteristics are reversible airway obstruction and increased respiratory response to various stimuli. The main factors of airway obstruction include tracheal smooth muscle spasm, respiratory mucosal edema, increased respiratory mucus secretion, eosinophil infiltration, airway epithelial damage or desquamation, etc. Asthma belongs to the category of "asthma" in traditional Chinese medicine, and is mostly triggered by invasion of exogenous evil, improper diet, emotional disorders, and physical weakness and fatigue.

[0006] At present, asthma medications recommended by domestic and foreign guidelines are mostly bronchodilators, theophylline, cholinergic receptor antagonists, hormones, etc. Although they can improve symptoms and relieve the condition to a certain extent, they still cannot cure asthma. Moreover, asthma is a heterogeneous disease with multiple clinical phenotypes, and its treatment is quite difficult. Summary of the invention

[0007] One or more embodiments of the present application provide a compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated form thereof.

[0008]

[0009] in

[0010] R is

[0011] Ring A is a 5- to 10-membered heteroaryl group containing 1-3 heteroatoms selected from N, O, and S, wherein at least one heteroatom is N.

[0012] Ring B is

[0013] X 1 , Y 1 are each independently N or NH, O or S, and X 1 , Y 1 At least one of them is N or NH;

[0014] R 1 , R 2 , R 3 Each is independently H, C1-C6 alkyl or C1-C6 alkoxy;

[0015] n is 1 or 2;

[0016] *Indicates the connection location.

[0017] In one or more embodiments, the five- to ten-membered heteroaryl group is substituted with one or more (1, 2, 3 or 4) substituents selected from C1-C6 alkyl and C1-C6 alkoxy.

[0018] In one or more embodiments, the five-membered to ten-membered heteroaryl group is a monocyclic heteroaryl group or a condensed-ring heteroaryl group.

[0019] In one or more embodiments, Ring A is

[0020] In one or more embodiments, X 2 , Y 2 , Z 2 are each independently N or NH, O or S, and X 2 , Y 2 , Z 2 At least one of them is N or NH.

[0021] In one or more embodiments, Ring A is

[0022] In one or more embodiments, X 2 are each independently N or NH, Y 2 , Z 2 are each independently N or NH, O or S.

[0023] In one or more embodiments, Ring B is X 1 , Y 1 N or NH; R 2 It is H or C1-C6 alkyl.

[0024] In one or more embodiments, Ring B is

[0025] In one or more embodiments, the compound of the present application is

[0026]

[0027]

[0028] One or more embodiments of the present application provide an intermediate compound for preparing the compound of the present application, wherein the intermediate compound is

[0029]

[0030] One or more embodiments of the present application provide a method for preparing the compound of the present application, which comprises (1)

[0032]

[0033] a) reacting the compound of formula II with sodium acetate at 80°C-120°C for 1-2 hours, adding the compound of formula I, and reacting at 20°C-50°C for 1-8 hours;

[0034] b) hydrolyzing the compound of formula III obtained in step a) at 50° C.-90° C. for 1-8 hours;

[0035] c) reacting the product compound of formula IV obtained in step b) with R B环 -NH 2 The reaction was carried out at 20°C-50°C for 5-7 hours to obtain a compound of formula V; (2)

[0037]

[0038] d) Compound VI, compound VII and K 2 CO 3 Stir at 20°C-50°C for 20-30 hours, then stir at -15°C until the solution turns orange-yellow, then add DBU and bromochloromethane at -20°C-0°C, and react at 20°C-50°C for 6-24 hours;

[0039] e) stirring the product compound of formula VIII obtained in step d) at 20° C.-50° C. for 8-12 hours for hydrolysis reaction;

[0040] f) reacting the product compound of formula IX obtained in step e) with R B环 -NH 2 React at 20℃-50℃ for 4-8 hours; (3)

[0042]

[0043] g) reacting the compound of formula XI with the compound of formula XII at 60° C.-100° C. for 10-14 hours;

[0044] h) hydrolyzing the compound of formula XIII obtained in step g) at 20° C.-50° C. for 2-6 hours;

[0045] i) reacting the compound of formula XIV obtained in step h) with R B环 -NH 2 React at 20°C-50°C for 10-14 hours;

[0046] or (4)

[0048]

[0049] j) stirring the compound of formula XVI and the compound of formula XVII at 60° C.-100° C. for 10-14 hours;

[0050] k) stirring the product compound of formula XVIII obtained in step j) at 60° C.-100° C. for hydrolysis reaction for 2-6 hours;

[0051] l) reacting the product compound of formula XIX obtained in step k) with R B环 -NH 2 Stir the reaction at 20°C-50°C for 10-14 hours;

[0052] Wherein, R is as described in any one of claims 1 to 5, R B环 is the B ring as described in any one of claims 1 to 5, and X is O or S.

[0053] One or more embodiments of the present application provide a pharmaceutical composition comprising a compound of the present application and a pharmaceutically acceptable auxiliary material or excipient.

[0054] One or more embodiments of the present application provide use of the compound of the present application or the pharmaceutical composition of the present application in the preparation of a medicament for preventing and / or treating inflammatory diseases, respiratory diseases, skin diseases, or immune system diseases.

[0055] One or more embodiments of the present application provide a compound of the present application for use as a medicament.

[0056] One or more embodiments of the present application provide the pharmaceutical composition of the present application, which is used as a medicament.

[0057] One or more embodiments of the present application provide a compound or composition of the present application for preventing and / or treating inflammatory diseases, respiratory diseases, skin diseases, or immune system diseases.

[0058] One or more embodiments of the present application provide a compound or composition of the present application for preventing and / or treating a disease mediated by PDE3 and / or PDE4.

[0059] One or more embodiments of the present application provide a compound or composition of the present application for inhibiting PDE3 or PDE4.

[0060] One or more embodiments of the present application provide a compound or composition of the present application for inhibiting PDE3 and PDE4.

[0061] One or more embodiments of the present application provide a method for preventing and / or treating inflammatory diseases, respiratory diseases, skin diseases, or immune system diseases, which comprises administering a compound or composition of the present application to a subject in need thereof.

[0062] One or more embodiments of the present application provide a method for preventing and / or treating a disease mediated by PDE3 and / or PDE4, the method comprising administering a compound or composition of the present application to a subject in need thereof.

[0063] One or more embodiments of the present application provide a method for inhibiting PDE3 and / or PDE4, the method comprising administering a compound or composition of the present application to a subject in need thereof.

[0064] In one or more embodiments, the inflammatory disease is an inflammatory skin disease or an inflammatory lung disease.

[0065] In one or more embodiments, the pulmonary inflammatory disease is asthma or chronic obstructive pulmonary disease.

[0066] In one or more embodiments, the respiratory disease is chronic obstructive pulmonary disease or asthma.

[0067] In one or more embodiments, the skin disease is psoriasis or atopic dermatitis.

[0068] In one or more embodiments, the immune system disease is systemic lupus erythematosus or rheumatoid arthritis.

[0069] One or more embodiments of the present application provide the use of the compound of the present application or the pharmaceutical composition of the present application in the preparation of a medicament for preventing and / or treating a disease mediated by PDE3 and / or PDE4, an inhibitor of PDE3 or PDE4, or an inhibitor of PDE3 and PDE4.

[0070] The following is an explanation of the terms used in the technical solution of the present application. As used in the specification and the appended claims, unless otherwise specifically stated, the terms of the present application have the following meanings:

[0071] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0072] The term "amino" refers to -NH 2 .

[0073] The term "hydroxy" refers to -OH.

[0074] "Alkyl" refers to a straight or branched saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, preferably an alkyl group of 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8) carbon atoms, more preferably an alkyl group of 1 to 6 carbon atoms, and further preferably an alkyl group of 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, neobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and various branched isomers thereof; when the alkyl group is substituted, it may be optionally further substituted by one or more substituents.

[0075] "Alkoxy" refers to a group formed by replacing at least one carbon atom in an alkyl group with an oxygen atom. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexyloxy, cyclopropyloxy and cyclobutyloxy. The definition of alkyl is the same as that of "alkyl" described above.

[0076] "Aryl" refers to a substituted or unsubstituted aromatic ring, which can be, for example, a monocyclic ring of 6 to 8 carbon atoms (e.g., 6, 7, 8 carbon atoms), a bicyclic ring of 6 to 12 carbon atoms (e.g., 6, 7, 8, 9, 10, 11, 12 carbon atoms), or a tricyclic ring system of 10 to 14 carbon atoms (e.g., 10, 11, 12, 13, 14 carbon atoms), which can be a bridged ring or a spirocyclic ring, and non-limiting examples include phenyl and naphthyl. The aryl group can be optionally further substituted by one or more substituents.

[0077] "Heteroaryl" refers to a substituted or unsubstituted aromatic ring, which can be, for example, a 5- to 8-membered (e.g., 5, 6, 7, 8-membered) monocyclic ring, a 5- to 12-membered (e.g., 5, 6, 7, 8, 9, 10, 11, 12-membered) bicyclic ring, and contains 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) heteroatoms selected from N, O or S, preferably a 5- to 8-membered heteroaryl, and 1 to 4 (e.g., 1, 2, 3, 4) N, S selectively substituted in the ring of the heteroaryl can be oxidized to various oxidation states. The heteroaryl can be attached to a heteroatom or a carbon atom, and the heteroaryl can be a bridged ring or a spirocyclic ring, and non-limiting examples include cyclic pyridyl, furanyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinylbenzimidazolyl, benzopyridinyl, pyrrolopyridinyl. The heteroaryl group is optionally further substituted with one or more substituents.

[0078] "Heterocyclyl" or "heterocycle" refers to a saturated or unsaturated non-aromatic heterocycle, which can be, for example, a 5- to 10-membered (e.g., 5, 6, 7, 8, 9, 10-membered) monocyclic ring, a 5- to 12-membered (e.g., 5, 6, 7, 8, 9, 10, 11, 12-membered) bicyclic ring, or a 10- to 15-membered (e.g., 10, 11, 12, 13, 14, 15-membered) tricyclic ring system, and contains 1 to 4 (e.g., 1, 2, 3, 4) heteroatoms selected from N, O or S, preferably a 5- to 8-membered heterocyclyl. The 1 to 4 (e.g., 1, 2, 3, 4) N and S optionally substituted in the ring of the "heterocyclyl" or "heterocycle" can be oxidized to various oxidation states; the "heterocyclyl" or "heterocycle" can be attached to a heteroatom or a carbon atom; the "heterocyclyl" or "heterocycle" can be a bridged ring or a spirocycle. Non-limiting examples of “heterocyclyl” or “heterocycle” include oxirane, oxirane, aziridine, oxetanyl, azetidinyl, thietanyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxhexacyclyl, azepanyl, oxepinyl, thiepanyl, oxazepinyl, diazepinyl, thiazepinyl, pyridinyl, piperidinyl, homopiperidinyl, furanyl, pyridinyl, piperidinyl, homopiperidinyl, furanyl, pyridinyl, piperidinyl, piperidinyl, homopiperidinyl, furanyl, pyridinyl, piperidinyl, piperidinyl, homopiperidinyl, pyridinyl, piperidinyl, piperidinyl, py ... pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, pyridinyl, py pyranyl, thiophene, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, piperidinyl, morpholinyl, thiomorpholinyl, thioxanyl, 1,3-dithianyl, dihydrofuranyl, dithiolanyl, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, oxazolyl, benzopyridyl, pyrrolopyridyl, benzodihydrofuranyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxane, 1,3-dioxolane, pyrazolinyl, dithianyl, dithiolanyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3 .1.0]hexyl, 3-azabicyclo[4.1.0]heptyl, azabicyclo[2.2.2]hexyl, 3H-indolylquinolizinyl, N-pyridylurea, 1,1-dioxothiomorpholinyl, azabicyclo[3.2.1]octanyl, azabicyclo[5.2.0]nonanyl, oxatricyclo[5.3.1.1]dodecyl, azaadamantyl and oxaspiro[3.3]heptanyl. The "heterocyclyl" or "heterocycle" may be optionally further substituted by one or more substituents.

[0079] "Cycloalkyl" refers to a saturated cyclic hydrocarbon group, which can be, for example, a monocyclic, bicyclic or polycyclic ring of 6 to 10 carbon atoms (e.g., 6, 7, 8, 9, 10 carbon atoms), preferably 6 to 8 carbon atoms. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. When the cycloalkyl is substituted, it can be optionally further substituted by one or more substituents.

[0080] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof" refers to a salt of the compound of the present invention that retains the biological effectiveness and properties of the free acid or free base, and the free acid is obtained by reacting with a non-toxic inorganic base or organic base, and the free base is obtained by reacting with a non-toxic inorganic acid or organic acid.

[0081] "Pharmaceutical composition" refers to a mixture of one or more compounds described herein, their pharmaceutically acceptable salts or prodrugs and other chemical components, wherein "other chemical components" refers to pharmaceutically acceptable carriers, excipients and / or one or more other therapeutic agents.

[0082] "Carrier" refers to a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0083] "Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

[0084] "Prodrug" refers to a compound of the present invention that can be converted into a biologically active compound through in vivo metabolism. The prodrug of the present invention is prepared by modifying the amino or carboxyl group in the compound of the present invention, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. When the prodrug of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free amino or carboxyl group.

[0085] "Co-crystal" refers to a crystal formed by the active pharmaceutical ingredient (API) and the co-crystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds, in which the pure state of API and CCF are solid at room temperature and there is a fixed stoichiometric ratio between the components. Co-crystal is a multi-component crystal, including binary eutectics formed between two neutral solids and multi-component eutectics formed between neutral solids and salts or solvates.

[0086] "Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformational isomers.

[0087] "Optional" or "optionally" or "selective" or "selectively" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group may but need not be present, and the description includes instances where the heterocyclyl group is substituted with alkyl group and instances where the heterocyclyl group is not substituted with alkyl group.

[0088] One or more embodiments of the present application provide a difluoromethoxyphenyl derivative, which is an inhibitor of PDE3 and / or PDE4 that can treat asthma.

[0089] In one or more embodiments, Formula I is R 5 for

[0090] In one or more embodiments, Formula I is R 6 for

[0091] In one or more embodiments, Formula I is R 7 for

[0092] In one or more embodiments, Formula I is R 8 for

[0093] In one or more embodiments, Formula I is R 9 for

[0094] In one or more embodiments, Formula I is R 10 for

[0095] In one or more embodiments, Formula I is R 11 for

[0096] In one or more embodiments, Formula I is R 12 for BRIEF DESCRIPTION OF THE DRAWINGS

[0097] Figure 1 It shows the effect of active compounds on the expression level of inflammatory factor IL-1β in Raw264.7 cells induced by LPS.

[0098] Figure 2It shows the effect of active compounds on the expression level of inflammatory factor IL-6 in Raw264.7 cells induced by LPS. Specific embodiments

[0099] The present invention will be further described below in conjunction with specific embodiments. These embodiments are only for the purpose of explanation and are not intended to limit the scope and essence of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present invention.

[0100] Example 1 Synthesis of Series A Compounds

[0101] (1) Synthesis of Compound 2

[0102]

[0103] Weigh 3g of compound 3,4-dihydroxybenzaldehyde (21.72mmol, 1.0equiv.) and 6.91g of sodium carbonate (65.16mmol, 3.0equiv.), dissolve in 30mL N,N-dimethylformamide, and finally add 3.44g of ethyl difluorochloroacetate (21.72mmol, 1.0equiv.), react at 80°C for 8 hours, detect the complete reaction of the raw materials by TLC, and add water to quench the reaction. Use ethyl acetate to extract 3 times, and wash the organic phase with saturated sodium chloride solution. The organic phase is concentrated and 1.4g of white solid product (compound 2) is obtained by rapid column chromatography, with a yield of 34%.

[0104] The obtained compound 2 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (500 MHz, CDCl 3 )δ9.90(s,1H),7.53(d,J=2.0Hz,1H),7.44(dd,J=8.5,2.0Hz,1H),7.26(d,J=8.5Hz,1H),6.66(br,1H).ESI-HRMS m / z: calculated value is C 8 H 6 O 3 F 2 Na + [M+Na] + , 211.0177; the measured value is 211.0166.

[0105] (2) Synthesis of Compound 3

[0106]

[0107] Weigh 2.1g of compound 2 (11.16mmol, 1.0equiv.), add 3.09g of potassium carbonate (22.32mmol, 2.0equiv.), add 20mL of N,N-dimethylformamide, and finally add 2.26g of bromomethylcyclopropane (16.74mmol, 1.5equiv.), react at 80°C for 8 hours, detect the complete reaction of the raw materials by TLC, and quench the reaction by adding water. Use ethyl acetate to extract 3 times, and wash the organic phase with saturated sodium chloride solution. Concentrate the organic phase and obtain 2.3g of yellow oily product (compound 3) by rapid column chromatography, with a yield of 85%.

[0108] The obtained compound 3 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (500 MHz, DMSO-d 6 )δ9.94(s,1H),7.59-7.54(m,2H),7.39(d,J=8.0Hz,1H),7.28(t,J=73.5Hz,1H),3.98(d,J=7.0Hz,2H),1.30-1.21(m,1H),0.61-0.55(m,2H),0.39-0.33(m,2H).ESI-MS m / z: calculated value is C 12 H 13 O 3 F 2 + [M+H] + , 243.1; the measured value is 243.1.

[0109] (3) Synthesis of Compound 4

[0110]

[0111] 3.21 g of methyl 2-amino-3-hydroxypropionate hydrochloride (20.64 mmol, 1.0 equiv.) was dissolved in 69 mL of N,N-dimethylformamide, and 8.55 g of potassium carbonate (61.92 mmol, 3.0 equiv.) was added. 2 After protection, 5.0 g of compound 3 (20.64 mmol, 1.0 equiv.) was added after stirring for 10 min, and the reaction was continued overnight at room temperature. After TLC monitoring, 12.28 g of bromochloroform (61.92 mmol, 3.0 equiv.) and 9.43 g of DBU (61.92 mmol, 3.0 equiv.) were added dropwise at -10 ° C. The reaction was continued for 24 h at room temperature. After TLC monitoring, water was added to quench the reaction. Ethyl acetate was used for extraction 3 times, and the organic phase was washed with a saturated sodium chloride solution. The organic phase was concentrated and subjected to rapid column chromatography to obtain 3.6 g of a white solid product (compound 4) with a yield of 51%.

[0112] The obtained compound 4 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (600 MHz, DMSO-d 6 )δ8.97 (s, 1H), 7.64-7.58 (m, 2H), 7.36 (s, 1H), 7.22 (t, J=73.8 Hz, 1H), 4.00 (d, J=7.2 Hz, 2H), 3.85 (s, 3H), 1.32-1.22 (m, 1H), 0.63-0.55 (m, 2H), 0.42-0.36 (m, 2H). ESI-HRMS m / z: calculated value is C 16 H 15 O 5 NF 2 Na + [M+Na] + , 362.0811; the measured value is 362.0822.

[0113] (4) Synthesis of Compound 5

[0114]

[0115] 3 g of compound 4 (8.84 mmol, 1.0 equiv.) was weighed and dissolved in 15 mL of a 1:1 mixed solvent of ethanol and water, and 2.12 g of sodium hydroxide (53.04 mmol, 6.0 equiv.) was added. The mixture was stirred at room temperature and monitored by TLC. After the reaction was complete, the ethanol was removed by rotary evaporation, an appropriate amount of water was added, and the pH was adjusted to 6 with dilute hydrochloric acid. The mixture was filtered, and the solid was washed with small amounts of water several times, and dried to obtain 2.8 g of a yellow solid product (compound 5), with a yield of 97%.

[0116] The obtained compound 5 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, CDCl 3 )δ8.39 (s, 1H), 7.71 (d, J = 2.0 Hz, 1H), 7.66 (dd, J = 8.4, 2.0 Hz, 1H), 7.26 (t, J = 8.4 Hz, 1H), 6.72 (t, J = 74.8 Hz, 1H), 3.96 (d, J = 6.8 Hz, 2H), 1.38-1.28 (m, 1H), 0.71-0.64 (m, 2H), 0.40-0.35 (m, 2H). ESI-MS m / z: calculated value is C 15 H 14 O 5 NF 2 + [M+H] +, 326.1; the measured value is 326.1.

[0117] (5) Synthesis of Compounds A1-A6

[0118]

[0119] 50 mg of compound 5 (0.15 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 59 mg of HATU (0.15 mmol, 1.0 equiv.) and 60 mg of DIPEA (0.45 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.15 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain products A1-A6; the compounds A1-A6 are as follows:

[0120]

[0121] Example 2 Synthesis of Compound A1

[0122] The synthesis steps were as in Example 1, and the amine reagent in the last step was 6-aminoindolone to obtain compound A1 with a yield of 55%. 1 H NMR (500 MHz, DMSO-d 6 )δ10.44(s, 1H), 10.08(s, 1H), 8.87(s, 1H), 7.73(d, J=2.0Hz, 1H), 7.68(d d, J=8.5, 2.0Hz, 1H), 7.52 (s, 1H), 7.38 (d, J=8.0Hz, 1H), 7.31 (dd, J=8.0, 2 .0Hz, 1H), 7.22 (d, J=74.0Hz, 1H), 7.16 (d, J=8.0Hz, 1H), 4.02 (d, J=7.0Hz, 2H), 3.43(s, 2H), 1.33-1.24(m, 1H), 0.64-0.58(m, 2H), 0.43-0.36(m, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ176.7, 159.9, 158.4, 150.2, 143.9, 143.1, 142.1, 137.7, 137.4, 124.3, 124.1, 121.3, 121.2, 119.3 (d, J = 256.5 Hz), 116.5, 113.1, 112.0, 102.0, 73.4, 35.5, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 23 H 19 O 5 N 3 F 2 Na + [M+Na] + , 478.1185; the measured value is 478.1186.

[0123] Example 3 Synthesis of Compound A2

[0124] The synthesis steps were as in Example 1. The amine reagent in the last step was 5-aminoindolin-2-one to obtain compound A2 with a yield of 50%. 1 H NMR (400 MHz, DMSO-d 6 )δ10.35(s, 1H), 9.99(s, 1H), 8.83(s, 1H), 7.73(d, J=2.0Hz, 1H), 7.69(s, 1H), 7.67 (dd, J=8.0, 1.6Hz, 1H), 7.55 (dd, J=8.4, 2.0Hz, 1H), 7.38 (d, J=8. 4Hz, 1H), 7.23 (t, J=74.0Hz, 1H), 6.80 (d, J=8.4Hz, 1H), 4.02 (d, J=8.0Hz, 2H), 3.50(s, 2H), 1.32-1.26(m, 1H), 0.65-0.54(m, 2H), 0.43-0.34(m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ176.5, 160.0, 158.3, 150.2, 143.0, 142.1, 140.1, 137.5, 132.2, 126.1, 124.2, 121.2, 120.3, 119.3, 118.0, 116.6 (t, J = 257.0 Hz), 111.9, 108.9, 73.4, 36.1, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 23 H 19 O 5 N 3 F 2 Na + [M+Na]+ , 478.1185; the measured value is 478.1189.

[0125] Example 4 Synthesis of Compound A3

[0126] The synthesis steps were as in Example 1. The amine reagent in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound A3 with a yield of 66%. 1 H NMR (500 MHz, DMSO-d 6 ) δ10.16 (s, 1H), 10.07 (s, 1H), 8.86 (s, 1H), 7.73 (d, J = 2.0Hz, 1H), 7.68 (dd, J = 8.4, 2. 0Hz, 1H), 7.44 (d, J=2.1Hz, 1H), 7.38 (d, J=8.3Hz, 1H), 7.28 (dd, J=8.1, 2.1Hz, 1H), 7.2 3(d, J=74.5Hz, 1H), 7.13 (d, J=8.2Hz, 1H), 4.02 (d, J=7.0Hz, 2H), 2.84 (t, J=7.5Hz, 2H) , 2.44 (dd, J=8.5, 6.6Hz, 2H), 1.33-1.26 (m, 1H), 0.64-0.58 (m, 2H), 0.42-0.37 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ170.3, 159.9, 158.4, 150.2, 143.1, 142.1, 138.4, 137.4, 137.1, 127.7, 124.2, 121.2, 119.4, 119.3, 116.5 (d, J = 256.5 Hz), 114.4, 112.0, 107.7, 73.3, 30.6, 24.4, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 24 H 21 O 5 N 3 F 2 Na + [M+Na] + , 492.1341; the measured value is 492.1357.

[0127] Example 5 Synthesis of Compound A4

[0128] The synthesis steps were as in Example 1. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound A4 with a yield of 66%. 1 H NMR (400 MHz, DMSO-d6 )δ10.06(s, 1H), 10.00(s, 1H), 8.83(s, 1H), 7.73(s, 1H), 7.68(d, J=8.0Hz, 1H), 7.63 (s, 1H), 7.54 (m, 1H), 7.39 (d, J=8.0Hz, 1H), 7.23 (t, J=74.0Hz, 1H ), 6.83 (d, J=8.0Hz, 1H), 4.02 (d, J=8.0Hz, 2H), 2.87 (t, J=8.0Hz, 2H), 2.45 (t, J=8.0Hz, 2H), 1.40-1.30 (m, 1H), 0.65-0.55 (m, 2H), 0.44-0.34 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ170.5, 160.4, 158.7, 150.6, 143.5, 142.5, 137.9, 135.1, 133.0, 124.6, 124.2, 121.7, 120.9, 120.2, 119.7, 117.0 (t, J = 257.4 Hz), 115.4, 112.3, 73.8, 30.8, 25.5, 10.4, 3.6 (2×C). ESI-HRMS m / z: calculated value is C 24 H 21 O 5 N 3 F 2 Na + [M+Na] + , 492.1341; the measured value is 492.1351.

[0129] Example 6 Synthesis of Compound A5

[0130] The synthesis steps were as in Example 1. The amine reagent in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound A5 with a yield of 20%. 1 H NMR (500 MHz, DMSO-d 6)δ10.92 (s, 1H), 10.28 (s, 1H), 8.90 (s, 1H), 7.91 (d, J = 9.0Hz, 2H), 7.80 (d, J = 9.0Hz, 2H ), 7.74 (d, J=2.0Hz, 1H), 7.69 (dd, J=8.5, 2.0Hz, 1H), 7.39 (d, J=8.5Hz, 1H), 7.23 (d, J=7 4.0Hz, 1H), 4.02 (d, J=7.0Hz, 2H), 3.45-3.36 (m, 1H), 2.73-2.66 (m, 1H), 2.24 (d, J=17.0 Hz, 1H), 1.34-1.26 (m, 1H), 1.09 (d, J=7.0Hz, 3H), 0.64-0.57 (m, 2H), 0.43-0.36 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.3, 160.0, 158.6, 152.2, 150.2, 143.4, 142.1, 139.3, 137.3, 130.3, 126.2 (2×C), 124.1, 121.2, 120.3 (2×C), 119.3, 116.5 (d, J=258.0 Hz), 112.0, 73.4, 33.5, 27.0, 15.9, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 26 H 25 O 5 N 4 F 2 + [M+H] + , 511.1788; the measured value is 511.1767.

[0131] Example 7 Synthesis of Compound A6

[0132] The synthesis steps were as in Example 1. The amine reagent in the last step was 6-(4-aminophenyl)-3(2H)-pyridazinone to obtain compound A6 with a yield of 43%. 1 H NMR (500 MHz, DMSO-d 6)δ13.15(s, 1H), 10.31(s, 1H), 8.91(s, 1H), 8.04(d, J=10.0Hz, 1H), 7.98-7. 92 (m, 2H), 7.90-7.84 (m, 2H), 7.74 (d, J = 2.0Hz, 1H), 7.69 (dd, J = 8.5, 2.0Hz, 1H), 7.9 (d, J=8.0Hz, 1H), 7.16 (d, J=74.0Hz, 1H), 6.99 (d, J=10.0Hz, 1H), 4. 02(d, J=7.0Hz, 2H), 1.33-1.25(m, 1H), 0.64-0.59(m, 2H), 0.43-0.38(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ160.2, 160.0, 158.7, 150.2, 143.5, 143.4, 142.1, 139.2, 137.3, 131.3, 130.1, 130.1, 126.1 (2×C), 124.1, 121.2, 120.6 (2×C), 119.3, 116.5 (d, J=256.5 Hz), 112.0, 73.4, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 25 H 21 O 5 N 2 F 2 Na + [M+Na] + , 495.1475; the measured value is 495.1491.

[0133] Example 8 Synthesis of Series B Compounds

[0134] (1) Synthesis of Compound 6

[0135]

[0136] Weigh 4.46g of 3,3-dibromo-1.1.1-trifluoro-2-one (16.52mmol, 2.0equiv.) and 2.7g of sodium acetate (33.04mmol, 4.0equiv.) into a reaction bottle, add an appropriate amount of water to dissolve, stir at 100°C for 1 hour, cool to room temperature, add 2g of compound 3 (8.26mmol, 1.0equiv.) in methanol and 4mL of ammonia water to the above solution, stir at room temperature, and when TLC detects that the raw material is no longer reduced, remove methanol by rotary evaporation and add water. Extract with ethyl acetate 3 times, wash the organic phase once with saturated sodium chloride solution, and dry the organic phase with anhydrous sodium sulfate. Concentrate the organic phase and obtain 1.37g of yellow solid product (compound 6) by rapid column chromatography, with a yield of 21%.

[0137] The obtained compound 6 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, CDCl 3 )δ12.56 (br, 1H), 7.50 (d, J = 2.0 Hz, 1H), 7.41 (s, 1H), 7.32 (dd, J = 8.4, 2.0 Hz, 1H), 7.09 (d, J = 8.4 Hz, 1H), 6.62 (t, J = 75.2 Hz, 1H), 3.72 (d, J = 7.2 Hz, 2H), 1.28-1.15 (m, 1H), 0.65-0.56 (m, 2H), 0.32-0.24 (m, 2H). ESI-MS m / z: calculated value is C 15 H 14 O 2 N 2 F 5 + [M+H] + , 349.1; the measured value is 349.1.

[0138] (2) Synthesis of Compound 7

[0139]

[0140] The synthesis steps were similar to those of compound 5, and a yellow solid product was obtained with a yield of 83%.

[0141] The obtained compound 7 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, DMSO-d 6)δ13.28 (br, 1H), 7.96-7.56 (m, 3H), 7.27 (d, J=8.4 Hz, 1H), 7.14 (t, J=74.4 Hz, 1H), 3.98 (d, J=6.8 Hz, 2H), 1.35-1.22 (m, 1H), 0.64-0.55 (m, 2H), 0.42-0.34 (m, 2H). ESI-HRMS m / z: calculated value is C 15 H 14 O 4 N 2 F 2 Na + [M+Na] + , 347.0814; the measured value is 347.0800.

[0142] (3) Synthesis of Compounds B1-B5

[0143]

[0144] 50 mg of compound 7 (0.15 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 59 mg of HATU (0.15 mmol, 1.0 equiv.) and 60 mg of DIPEA (0.45 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.15 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds B1-B5; the compounds B1-B5 are as follows:

[0145]

[0146] Example 9 Synthesis of Compound B1

[0147] The synthesis steps were as in Example 8. The amine reagent in the last step was 6-aminoindolone to obtain compound B1 with a yield of 73%. 1 H NMR (600 MHz, DMSO-d 6)δ13.20(br,1H),10.32(s,1H),9.65(s,1H),7.92(s,1H),7.79(s,1H),7 .72 (s, 1H), 7.65 (d, J = 8.4Hz, 1H), 7.55 (d, J = 9.0Hz, 1H), 7.28 (d, J = 8.4H z, 1H), 7.13 (t, J=74.5Hz, 1H), 6.79 (d, J=8.4Hz, 1H), 3.99 (d, J=7.2Hz, 2 H), 3.49 (s, 2H), 1.34-1.27 (m, 1H), 0.63-0.58 (m, 2H), 0.40-0.36 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.5, 160.5, 150.2, 145.1, 140.2, 139.5, 137.0, 132.9, 128.2, 126.1, 121.7, 121.4, 119.6, 118.0, 117.4, 116.8 (d, J = 256.5 Hz), 111.5, 108.9, 73.3, 36.2, 10.1, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 23 H 20 O 4 N 4 F 2 Na + [M+Na] + , 477.1345; the measured value is 477.1341.

[0148] Example 10 Synthesis of Compound B2

[0149] The synthesis steps were as in Example 8. The amine reagent in the last step was 5-aminoindolin-2-one to obtain compound B2 with a yield of 42%. 1 H NMR (400 MHz, DMSO-d 6 )δ13.14(br, 1H), 10.34(s, 1H), 9.66(s, 1H), 7.96(s, 1H), 7.81(s, 1H), 7 .74 (s, 1H), 7.66 (d, J = 8.8Hz, 1H), 7.57 (d, J = 8.4Hz, 1H), 7.29 (d, J = 8.4H z, 1H), 7.15 (t, J=74.4Hz, 1H), 6.78 (d, J=8.4Hz, 1H), 3.98 (d, J=7.2Hz, 2 H), 3.50 (s, 2H), 1.37-1.27 (m, 1H), 0.65-0.55 (m, 2H), 0.43-0.33 (m, 2H). 13C NMR (101 MHz, DMSO-d 6 )δ176.4, 160.4, 150.1, 145.0, 140.1, 139.5, 137.0, 132.9, 128.2, 126.0, 121.7, 121.3, 119.5, 117.9, 117.3, 116.8 (t, J = 256.7 Hz), 111.3, 108.9, 73.2, 36.1, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 23 H 20 O 4 N 4 F 2 Na + [M+Na] + , 477.1345; the measured value is 477.1340.

[0150] Example 11 Synthesis of Compound B3

[0151] The synthesis steps were as in Example 8. The amine reagent in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound B3 with a yield of 41%. 1 H NMR (500 MHz, DMSO-d 6 )δ13.16(s, 1H), 10.12(s, 1H), 9.68(s, 1H), 7.96(s, 1H), 7.81(s, 1H), 7.6 7(d, J=8.0Hz, 1H), 7.50(s, 1H), 7.33-7.22(m, 2H), 7.14(t, J=74.5Hz, 1H), 7.11 (d, J=74.5Hz, 1H), 3.99 (d, J=7.0Hz, 2H), 2.83 (t, J=7.5Hz, 2H), 2.44 (t, J=7.5Hz, 2H), 1.36-1.24(m, 1H), 0.66-0.56(m, 2H), 0.44-0.34(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ170.4, 160.5, 150.1, 145.1, 140.1, 138.4, 137.7, 136.8, 128.1, 127.7, 121.9, 121.3, 118.6, 117.9, 116.7 (d, J = 256.5 Hz), 113.8, 111.4, 107.1, 73.2, 30.7, 24.4, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 24 H 23 O 4 N4 F 2 + [M+H] + , 469.1682; the measured value is 469.1681.

[0152] Example 12 Synthesis of Compound B4

[0153] The synthesis steps were as in Example 8. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound B4 with a yield of 83%. 1 H NMR (400 MHz, DMSO-d 6 )δ13.14(s, 1H), 10.05(s, 1H), 9.65(s, 1H), 7.95(s, 1H), 7.80(s, 1H), 7.66 (s, 2H), 7.55 (d, J=8.8Hz, 1H), 7.29 (d, J=8.4Hz, 1H), 7.15 (t, J=74.4Hz, 1H ), 6.81 (d, J = 8.4Hz, 1H), 3.98 (d, J = 6.8Hz, 2H), 2.86 (t, J = 7.6Hz, 2H), 2.44 (t, J=7.6Hz, 2H), 1.36-1.26 (m, 1H), 0.65-0.55 (m, 2H), 0.43-0.33 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ170.0, 160.4, 150.1, 145.1, 140.1, 136.9, 134.1, 133.2, 128.2, 123.7, 121.7, 121.3, 119.8, 119.0, 117.9, 116.7 (t, J = 256.3 Hz), 115.0, 111.3, 73.2, 30.5, 25.1, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 24 H 22 O 4 N 4 F 2 Na + [M+Na] + , 491.1501; the measured value is 491.1496.

[0154] Example 13 Synthesis of Compound B5

[0155] The synthesis steps were as in Example 8. The amine reagent used in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound B5 with a yield of 15%. 1H NMR (500 MHz, DMSO-d 6 )δ13.22(br, 1H), 10.91(s, 1H), 9.98(s, 1H), 8.00(s, 1H), 7.92(d, J=8.5Hz, 2H), 7.84(s , 1H), 7.78 (d, J=9.0Hz, 2H), 7.70 (dd, J=8.0, 2.0Hz, 1H), 7.29 (d, J=8.5Hz, 1H), 7.07 (t, J =74.5Hz, 1H), 4.00 (d, J = 7.0Hz, 2H), 3.44-3.37 (m, 1H), 2.74-2.64 (m, 1H), 2.24 (d, J = 16. 0Hz, 1H), 1.35-1.26 (m, 1H), 1.09 (d, J=7.0Hz, 3H), 0.64-0.59 (m, 2H), 0.42-0.36 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.3, 160.5, 152.3, 150.1, 145.3, 140.2, 139.9, 136.6, 129.5, 128.1, 126.2 (2×C), 122.3, 121.3, 119.7 (2×C), 118.1, 116.7 (d, J=256.5 Hz), 111.5, 73.2, 33.6, 27.0, 15.9, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 26 H 25 O 4 N 5 F 2 Na + [M+Na] + , 532.1767; the measured value is 532.1747.

[0156] Example 14 Synthesis of C series compounds

[0157] (1) Synthesis of Compound 8

[0158]

[0159] The synthesis steps were similar to those of compound 4. Compound 3 and D-cysteine ​​methyl ester hydrochloride were reacted to obtain a yellow solid product with a yield of 15%.

[0160] The obtained compound 8 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, CDCl 3)δ8.17 (s, 1H), 7.66 (d, J = 2.0 Hz, 1H), 7.46 (dd, J = 8.4, 2.0 Hz, 1H), 7.22 (d, J = 8.0 Hz, 1H), 6.70 (t, J = 75.2 Hz, 1H), 4.00-3.98 (m, 5H), 1.38-1.27 (m, 1H), 0.70-0.64 (m, 2H), 0.41-0.36 (m, 2H). ESI-HRMS m / z: calculated value is C 16 H 15 O 4 NF 2 SNa + [M+Na] + , 378.0582; the measured value is 378.0581.

[0161] (2) Synthesis of Compound 9

[0162]

[0163] The synthesis steps were similar to those of compound 5, and a yellow solid product was obtained with a yield of 84%.

[0164] The obtained compound 9 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (500 MHz, DMSO-d 6 )δ8.18(s,1H),7.63(s,1H),7.51(d,J=8.5Hz,1H),7.28(d,J=8.0Hz,1H),7.18(t,J=74.0Hz,1H),4.01(d,J=7.0Hz,2H),1.32-1.24(m,1H),0.62-0.53(m,2H),0.45-0.33(m,2H),OH(not observed).ESI-MS m / z:calculated value is C 15 H 14 O 4 NF 2 S + [M+H] + , 342.1; the measured value is 342.1.

[0165] (3) Synthesis of Compounds C1-C7

[0166]

[0167] 50 mg of compound 9 (0.15 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 59 mg of HATU (0.15 mmol, 1.0 equiv.) and 60 mg of DIPEA (0.45 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.15 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction of the raw material after the reaction was complete, extracted with ethyl acetate, washed once with a saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds C1-C7; the compounds C1-C7 are as follows:

[0168]

[0169] Example 15 Synthesis of Compound C1

[0170] The synthesis steps were as in Example 14. The amine reagent in the last step was 6-aminoindolone to obtain compound C1 with a yield of 18%. 1 H NMR (600 MHz, DMSO-d 6 ) δ10.46 (s, 1H), 10.19 (s, 1H), 8.46 (s, 1H), 7.84 (s, 1H), 7.70 (d, J = 8.4Hz, 1H), 7.55 (s, 1H), 7.39-7.27 (m, 2H), 7.20 (d, J = 5. 4Hz, 1H), 7.18 (t, J=60.6Hz, 1H), 4.07 (d, J=6.6Hz, 2H), 3.45 (s, 2H), 1.36-1.25 (m, 1H), 0.65-0.55 (m, 2H), 0.45-0.33 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.8, 166.4, 158.9, 150.4, 150.3, 144.0, 141.8, 137.7, 130.6, 125.6, 124.4, 121.3, 121.2, 119.6, 116.6 (d, J = 256.5 Hz), 113.3, 112.4, 102.1, 73.4, 35.5, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 23 H 19 O 4 N 3 F 2 SNa + [M+Na] + , 494.0957; the measured value is 494.0966.

[0171] Example 16 Synthesis of Compound C2

[0172] The synthesis steps were as in Example 14. The amine reagent used in the last step was 5-aminoindolin-2-one to obtain compound C2 with a yield of 57%. 1 H NMR (400 MHz, DMSO-d 6 )δ10.39 (s, 1H), 10.14 (s, 1H), 8.44 (s, 1H), 7.84 (d, J=2.0Hz, 1H), 7.73 (s , 1H), 7.70 (dd, J=8.4, 2.0Hz, 1H), 7.59 (dd, J=8.4, 2.0Hz, 1H), 7.33 (d, J=8 .4Hz, 1H), 7.22 (t, J=74.4Hz, 1H), 6.82 (d, J=8.4Hz, 1H), 4.06 (d, J=6.8Hz, 2H), 3.52(s, 2H), 1.36-1.30(m, 1H), 0.65-0.54(m, 2H), 0.45-0.34(m, 2H). 13 CNMR (101 MHz, DMSO-d 6 )δ176.4, 166.3, 158.7, 150.6, 150.3, 141.8, 140.2, 132.2, 130.6, 126.1, 125.3, 121.3, 120.4, 119.6, 118.1, 116.6 (t, J = 257.2 Hz), 112.3, 108.9, 73.4, 36.1, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 23 H 19 O 4 N 3 F 2 SNa + [M+Na] + , 494.0957; the measured value is 494.0965.

[0173] Example 17 Synthesis of Compound C3

[0174] The synthesis steps were as in Example 14. The amine reagent used in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound C3 with a yield of 59%. 1 H NMR (400 MHz, DMSO-d 6) δ10.20 (s, 2H), 8.47 (s, 1H), 7.84 (d, J = 2.0Hz, 1H), 7.70 (dd, J = 8.4, 2.0Hz, 1H), 7.50 (d, J=2.0Hz, 1H), 7.34 (d, J=8.4Hz, 1H), 7.29 (dd, J=8.4, 2.4Hz, 1H), 7.22 (t, J=74.4Hz, 1H), 7.16 (d, J=8.4Hz, 1H), 4.06 (d, J=7.2Hz, 2H), 2.85 (t, J=7.6Hz, 2H ), 2.45 (t, J=7.6Hz, 2H), 1.36-1.29 (m, 1H), 0.66-0.56 (m, 2H), 0.43-0.36 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ170.4, 166.4, 158.9, 150.4, 150.3, 141.8, 138.5, 137.2, 130.6, 127.8, 125.6, 121.3, 119.6, 119.5, 116.6 (t, J = 256.7 Hz), 114.6, 112.3, 107.8, 73.4, 30.6, 24.4, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 24 H 21 O 4 N 3 F 2 SNa + [M+Na] + , 508.1113; the measured value is 508.1133.

[0175] Example 18 Synthesis of Compound C4

[0176] The synthesis steps were as in Example 14. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound C4 with a yield of 53%. 1 H NMR (500 MHz, DMSO-d 6)δ10.11 (s, 1H), 10.08 (s, 1H), 8.43 (s, 1H), 7.82 (d, J=2.0Hz, 1H), 7.70 (dd, J=8.5, 2 .0Hz, 1H), 7.67 (d, J=2.0Hz, 1H), 7.57 (dd, J=8.5, 2.5Hz, 1H), 7.33 (d, J=8.5Hz, 1H), 7 .13 (d, J=74.0Hz, 1H), 6.85 (d, J=8.5Hz, 1H), 4.06 (d, J=7.0Hz, 2H), 2.89 (t, J=7.5Hz , 2H), 2.46 (t, J=7.5Hz, 2H), 1.35-1.24 (m, 1H), 0.68-0.54 (m, 2H), 0.46-0.33 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ170.0, 166.3, 158.7, 150.5, 150.3, 141.8, 134.7, 132.5, 130.6, 125.2, 123.7, 121.3, 120.6, 119.9, 119.6, 116.6 (d, J = 256.5 Hz), 115.0, 112.4, 73.4, 30.4, 25.1, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 24 H 21 O 4 N 3 F 2 SNa + [M+Na] + , 508.1113; the measured value is 508.1168.

[0177] Example 19 Synthesis of Compound C5

[0178] The synthesis steps were as in Example 14. The amine reagent used in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound C5 with a yield of 70%. 1 H NMR (500 MHz, DMSO-d 6)δ10.94 (s, 1H), 10.36 (s, 1H), 8.46 (s, 1H), 7.95-7.89 (m, 2H), 7.82 (d, J=2.0Hz, 1H), 7. 82-7.77 (m, 2H), 7.67 (dd, J=8.0, 2.0Hz, 1H), 7.31 (d, J=8.5Hz, 1H), 7.09 (t, J=74.0Hz, 1H ), 4.04 (d, J=6.5Hz, 2H), 3.44-3.35 (m, 1H), 2.68 (dd, J=17.0, 7.0Hz, 1H), 2.24 (d, J=17. 0Hz, 1H), 1.31-1.23 (m, 1H), 1.08 (d, J=7.0Hz, 3H), 0.62-0.57 (m, 2H), 0.41-0.36 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.7 (2×C), 159.3, 152.6, 150.5, 150.3, 142.0, 139.4, 130.7, 130.5, 126.4 (2×C), 126.1, 121.5, 120.7 (2×C), 119.8, 116.7 (t, J=256.5 Hz), 112.5, 73.6, 33.7, 27.2, 16.1, 10.2, 3.3 (2×C). ESI-HRMS m / z: calculated value is C 26 H 25 O 4 N 4 F 2 S + [M+H] + , 527.1559; the measured value is 527.1542.

[0179] Example 20 Synthesis of Compound C6

[0180] The synthesis steps were as in Example 14. The amine reagent used in the last step was 6-(4-aminophenyl)-3(2H)-pyridazinone to obtain compound C6 with a yield of 49%. 1 H NMR (600 MHz, DMSO-d 6) δ13.16 (s, 1H), 10.38 (s, 1H), 8.52 (s, 1H), 8.05 (d, J = 9.6Hz, 1H), 7.99 ( d, J=8.4Hz, 2H), 7.94-7.88 (m, 2H), 7.84 (s, 1H), 7.72 (d, J=8.4Hz, 1H), 7. 34(d, J=7.8Hz, 1H), 7.21(t, J=74.2Hz, 1H), 6.99(d, J=9.8Hz, 1H), 4.07( d, J=7.1Hz, 2H), 1.33-1.27(m, 1H), 0.64-0.58(m, 2H), 0.43-0.38(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.5, 160.2, 159.1, 150.3, 150.2, 143.5, 141.8, 139.3, 131.3, 130.6, 130.1, 126.1 (2×C), 125.9, 121.3, 120.7 (2×C), 120.2, 119.6, 116.6 (t, J=257.0 Hz), 112.4, 73.4, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 25 H 20 O 4 N 4 F 2 SNa + [M+Na] + , 533.1066; the measured value is 533.1052.

[0181] Example 21 Synthesis of Compound C7

[0182] The synthesis steps were as in Example 14. The amine reagent used in the last step was 6-amino-3(2H)-pyridazinone to obtain compound C7 with a yield of 35%. 1 H NMR (400 MHz, DMSO-d 6 ) δ12.91 (s, 1H), 10.61 (s, 1H), 8.55 (s, 1H), 7.94 (d, J = 10.0Hz, 1H), 7.85 (d, J = 2.0Hz, 1H), 7.68 (dd, J = 8.4, 2.0Hz, 1H), 7.32 (d, J = 8.4H z, 1H), 7.21 (t, J=74.0Hz, 1H), 6.99 (d, J=10.0Hz, 1H), 4.05 (d, J=6.8Hz, 2H), 1.32-1.26 (m, 1H), 0.64-0.55 (m, 2H), 0.43-0.34 (m, 2H). 13C NMR (101 MHz, DMSO-d 6 )δ166.6, 160.0, 159.6, 150.3, 149.1, 141.8, 140.7, 130.9, 130.8, 130.5, 126.7, 121.3, 119.6, 116.6 (t, J = 257.0 Hz), 112.2, 73.3, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 19 H 16 O 4 N 4 F 2 SNa + [M+Na] + , 457.0753; the measured value is 457.0740.

[0183] Example 22 Synthesis of D series compounds

[0184] (1) Synthesis of Compound 10

[0185]

[0186] 150 mg of compound 3 (0.62 mmol, 1.0 equiv.) was dissolved in 1.5 mL of N,N-dimethylformamide, and then 103 mg of methyl 2,3-diaminobenzoate (0.62 mmol, 1.0 equiv.) and 141 mg of sodium pyrosulfite (0.74 mmol, 1.2 equiv.) were added, and the mixture was heated to 80°C for overnight reaction and monitored by thin layer chromatography. After the reaction, water was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was dried over anhydrous sodium sulfate, and the solution was concentrated. 151 mg of compound 10 was obtained by column chromatography with a yield of 63%.

[0187] The obtained compound 10 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (500 MHz, CDCl 3 )δ10.71(br, 1H), 7.87(d, J=8.0 Hz, 1H), 7.76(d, J=7.5 Hz, 1H), 7.59(s, 1H), 7.34(d, J=8.5 Hz, 1H), 7.19(t, J=8.5 Hz, 1H), 7.09(d, J=8.5 Hz, 1H), 6.65(t, J=75.0 Hz, 1H), 3.88(s, 3H), 3.82(d, J=7.0 Hz, 2H), 1.23-1.20(m, 1H), 0.59-0.55(m, 2H), 0.29-0.26(m, 2H).ESI-MS m / z: calculated value is C 20 H19 O 4 N 2 F 2 + [M+H] + , 389.1; the measured value is 389.1.

[0188] (2) Synthesis of Compound 11

[0189]

[0190] The synthesis steps were similar to those of compound 5, and a white solid product was obtained with a yield of 87%.

[0191] The obtained compound 11 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (400 MHz, CD 3 OD) δ 7.89-7.84 (m, 2H), 7.76-7.72 (m, 2H), 7.30-7.26 (m, 2H), 6.87 (t, J = 75.2 Hz, 1H), 4.06 (d, J = 6.8 Hz, 2H), 1.42-1.35 (m, 1H), 0.69-0.65 (m, 2H), 0.43-0.41 (m, 2H). ESI-HRMS m / z: calculated value is C 19 H 17 O 4 N 2 F 2 + [M+H] + , 375.1151; the measured value is 375.1160.

[0192] (3) Synthesis of Compounds D1-D7

[0193]

[0194] 50 mg of compound 11 (0.13 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 51 mg of HATU (0.13 mmol, 1.0 equiv.) and 52 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.13 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds D1-D7; the compounds D1-D7 are as follows:

[0195]

[0196] Example 23 Synthesis of Compound D1

[0197] The synthesis steps were as in Example 22. The amine reagent in the last step was 6-aminoindolone to obtain compound D1 with a yield of 81%. 1 H NMR (400 MHz, DMSO-d 6 ) δ13.57 (s, 1H), 12.12 (s, 1H), 10.39 (s, 1H), 8.01 (d, J = 2.0Hz, 1H), 7.96 (dd, J = 7.6, 1.2Hz , 1H), 7.89 (dd, J=8.4, 2.0Hz, 1H), 7.85 (d, J=2.0Hz, 1H), 7.79 (dd, J=8.0, 0.8Hz, 1H), 7.60 (dd, J=8.4, 2.0Hz, 1H), 7.46-7.40 (m, 2H), 7.25 (t, J=74.4Hz, 1H), 6.84 (d, J=8.4Hz, 1H), 4 .10(d, J=6.8Hz, 2H), 3.54(s, 2H), 1.42-1.31(m, 1H), 0.68-0.58(m, 2H), 0.47-0.38(m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ176.3, 162.4, 151.3, 150.1, 141.6, 140.7, 139.8, 135.2, 133.2, 126.8, 126.6, 123.0, 122.9, 122.0, 121.3, 119.5, 118.5, 116.6 (t, J = 257.0 Hz), 116.5, 115.5, 112.4, 109.2, 73.2, 36.2, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated for C 27 H 22 O 4 N 4 F 2 Na + [M+Na] + , 527.1501; the measured value is 527.1495.

[0198] Example 24 Synthesis of Compound D2

[0199] The synthesis steps were as in Example 22. The amine reagent used in the last step was 5-aminoindolin-2-one to obtain compound D2 with a yield of 79%. 1 H NMR (500 MHz, DMSO-d 6)δ13.56 (s, 1H), 12.25 (s, 1H), 10.44 (s, 1H), 8.00 (d, J=2.0Hz, 1H), 7.97 (d, J= 7.5Hz, 1H), 7.89 (dd, J=8.5, 2.0Hz, 1H), 7.83-7.78 (m, 2H), 7.47-7.40 (m, 2H), 7.24 (d, J=74.0Hz, 1H), 7.19 (d, J=8.0Hz, 1H), 7.11-7.05 (m, 1H), 4.09 (d, J=7. 0Hz, 2H), 3.47(s, 2H), 1.41-1.32(m, 1H), 0.68-0.62(m, 2H), 0.45-0.38(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.8, 162.7, 151.3, 150.1, 144.4, 141.6, 140.7, 138.5, 135.2, 126.8, 124.6, 123.1, 122.9, 121.8, 121.3, 120.7, 119.5, 116.6 (d, J = 258.0 Hz), 115.7, 112.5, 111.7, 101.0, 73.2, 35.5, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated for C 27 H 23 O 4 N 4 F 2 + [M+H] + , 505.1682; the measured value is 505.1672.

[0200] Example 25 Synthesis of Compound D3

[0201] The synthesis steps were as in Example 22. The amine reagent used in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound D3 with a yield of 83%. 1 H NMR (400 MHz, DMSO-d 6)δ13.60 (s, 1H), 12.22 (s, 1H), 10.20 (s, 1H), 8.01 (d, J=2.0Hz, 1H), 7.98 (dd, J=7.6, 2.0Hz, 1H), 7.9 3 (dd, J=8.4, 2.0Hz, 1H), 7.82 (dd, J=8.0, 1.2Hz, 1H), 7.57 (d, J=2.0Hz, 1H), 7.45 (dd, J=8.0, 3.2Hz, 2H), 7.33 (dd, J=8.0, 2.0Hz, 1H), 7.26 (t, J=74.0Hz, 1H), 7.18 (d, J=8.0Hz, 1H), 4.09 (d, J=7.2Hz, 2H ), 2.88(t, J=8.0Hz, 2H), 2.49-2.43(m, 2H), 1.42-1.30(m, 1H), 0.68-0.59(m, 2H), 0.47-0.38(m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ170.4, 162.6, 151.4, 150.1, 141.6, 140.8, 138.9, 138.0, 135.3, 128.2, 126.7, 123.1, 123.0, 121.8, 121.4, 119.6, 119.0, 116.6 (t, J = 257.0 Hz), 115.7, 112.8, 112.4, 106.3, 73.2, 30.6, 24.4, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated for C 28 H 24 O 4 N 4 F 2 Na + [M+Na] + , 541.1658; the measured value is 541.1652.

[0202] Example 26 Synthesis of Compound D4

[0203] The synthesis steps were as in Example 22. The amine reagent used in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound D4 with a yield of 77%. 1 H NMR (500 MHz, DMSO-d 6)δ13.54(s, 1H), 12.12(s, 1H), 10.10(s, 1H), 8.02(s, 1H), 7.96(d, J=7.5Hz, 1H), 7.90(d , J=8.5Hz, 1H), 7.79 (d, J=8.0Hz, 1H), 7.75 (s, 1H), 7.60 (d, J=9.0Hz, 1H), 7.50-7.38 (m, 2H), 7.24 (t, J=74.0Hz, 1H), 6.91 (d, J=8.5Hz, 1H), 4.11 (d, J=7.0Hz, 2H), 2.94 (t, J=7.5 Hz, 2H), 2.48 (d, J=5.5Hz, 2H), 1.42-1.31 (m, 1H), 0.70-0.58 (m, 2H), 0.47-0.38 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ169.9, 162.4, 151.3, 150.1, 141.6, 140.7, 135.2, 134.4, 133.4, 126.7, 124.3, 123.0, 122.9, 121.9, 121.4, 119.5, 119.0, 118.2, 116.6 (d, J = 256.5 Hz), 115.5, 115.4, 112.5, 73.1, 30.4, 25.2, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated for C 28 H 24 O 4 N 4 F 2 Na + [M+Na] + , 541.1658; the measured value is 541.1647.

[0204] Example 27 Synthesis of Compound D5

[0205] The synthesis steps were as in Example 22. The amine reagent used in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound D5 with a yield of 27%. 1 H NMR (400 MHz, DMSO-d 6)δ13.59 (br, 1H), 12.43 (s, 1H), 10.97 (s, 1H), 8.04 (d, J = 2.0Hz, 1H), 7.98 (d, J = 7.6Hz, 1H) , 7.96-7.90 (m, 3H), 7.86 (s, 1H), 7.85-7.80 (m, 2H), 7.46-7.42 (m, 2H), 7.25 (t, J=74.4Hz, 1 H), 4.11 (d, J=7.2Hz, 2H), 3.42 (t, J=8.8Hz, 1H), 2.73 (dd, J=16.8, 6.8Hz, 1H), 2.26 (d, J=1 6.8Hz, 1H), 1.43-1.31 (m, 1H), 1.12 (d, J=7.2Hz, 3H), 0.69-0.58 (m, 2H), 0.50-0.37 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ166.4, 162.9, 152.1, 151.4, 150.1, 141.6, 140.9, 139.9, 135.4, 130.0 (2×C), 126.7, 126.6 (2×C), 123.2, 122.9, 121.6, 121.3, 119.5, 119.2, 116.6 (t, J=257.0 Hz), 115.9, 112.5, 73.2, 33.6, 27.0, 16.0, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 30 H 27 O 4 N 5 F 2 Na + [M+Na] + , 582.1923; the measured value is 582.1901.

[0206] Example 28 Synthesis of Compound D6

[0207] The synthesis steps were as in Example 22. The amine reagent used in the last step was 6-(4-aminophenyl)-3(2H)-pyridazinone to obtain compound D6 with a yield of 70%. 1 H NMR (500 MHz, DMSO-d 6)δ14.21(s, 1H), 13.17(s, 1H), 12.42(s, 1H), 8.19(d, J=2.0Hz, 1H), 8.05(d, J=10.0Hz, 1H), 8.03-7.96(m, 4H), 7.95-7.90(m, 2H), 7.83(d, J=8.0Hz, 1H), 7.45-7.40 (m, 2H), 7.25 (t, J=75.5Hz, 1H), 7.00 (dd, J=10.0, 2.0Hz, 1H), 4.1 5(d, J=7.0Hz, 2H), 1.41-1.32(m, 1H), 0.67-0.61(m, 2H), 0.46-0.41(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ163.0, 160.2, 151.5, 150.1, 143.4, 141.6, 140.8, 139.9, 135.4, 131.2, 130.2, 129.8, 126.8, 126.5 (2×C), 123.1, 122.8, 121.5, 121.3, 119.7, 119.5 (2×C), 116.6 (t, J=256.5 Hz), 115.9, 112.9, 73.3, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 29 H 23 O 4 N 5 F 2 Na + [M+Na] + , 566.1610; the measured value is 566.1592.

[0208] Example 29 Synthesis of Compound D7

[0209] The synthesis steps were as in Example 22. The amine reagent used in the last step was 6-amino-3(2H)-pyridazinone to obtain compound D7 with a yield of 18%. 1 H NMR (600 MHz, DMSO-d 6) δ12.83 (s, 1H), 12.40 (s, 1H), 8.34 (d, J = 10.2Hz, 1H), 7.97 (s, 2H), 7.84 (t, J = 8.4Hz, 2H), 7.45-7.38 (m, 2H), 7.22 (t, J = 7 4.4Hz, 1H), 7.02 (d, J=9.6Hz, 1H), 4.05 (d, J=6.6Hz, 2H), 1.36-1.30 (m, 1H), 0.64-0.60 (m, 2H), 0.44-0.40 (m, 2H), NH (not observed). 13 C NMR (151 MHz, DMSO-d 6 )δ163.4, 160.1, 151.7, 150.2, 141.9, 141.8, 141.0, 135.5, 131.1, 129.3, 126.9, 123.3, 122.9, 121.4, 120.5, 119.7, 116.7 (t, J = 256.8 Hz), 116.5, 112.9, 73.4, 10.1, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 23 H 19 O 4 N 5 F 2 Na + [M+Na] + , 490.1297; the measured value is 490.1284.

[0210] Example 30 Synthesis of E series compounds

[0211] (1) Synthesis of Compound 12

[0212]

[0213] The synthesis steps were similar to those of compound 10. Compound 3 and methyl 3,4-diaminobenzoate were reacted to obtain a yellow solid product with a yield of 71%.

[0214] The obtained compound 12 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (400 MHz, DMSO-d 6)δ13.27 (br, 1H), 8.20 (s, 1H), 7.90 (s, 1H), 7.85 (d, J = 8.4 Hz, 1H), 7.79 (dd, J = 8.4, 2.0 Hz, 1H), 7.68 (s, 1H), 7.37 (d, J = 8.4 Hz, 1H), 7.21 (t, J = 74.0 Hz, 1H), 4.02 (d, J = 7.2 Hz, 2H), 3.87 (s, 3H), 1.35-1.28 (m, 1H), 0.64-0.59 (m, 2H), 0.43-0.39 (m, 2H). ESI-HRMS m / z: calculated value is C 20 H 19 O 4 N 2 F 2 + [M+H] + , 389.1307; the measured value is 389.1323.

[0215] (2) Synthesis of Compound 13

[0216]

[0217] The synthesis steps were similar to those of compound 5, and a white solid product was obtained with a yield of 63%.

[0218] The obtained compound 13 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (500 MHz, DMSO-d 6 )δ8.20 (d, J=1.5Hz, 1H), 7.95 (d, J=2.0Hz, 1H), 7.85 (dd, J=8.5, 1.5Hz, 1H), 7.82 (dd, J=8.5, 2.0Hz, 1H), 7.66 (d, J=8.5Hz, 1H), 7. 36 (d, J=8.5Hz, 1H), 7.19 (t, J=74.5Hz, 1H), 4.03 (d, J=7.0Hz, 2H), 1.34-1.28 (m, 1H), 0.63-0.58 (m, 2H), 0.42-0.38 (m, 2H), OH (not observed), NH (not observed).ESI-HRMS m / z: The calculated value is C 19 H 17 O 4 N 2 F 2 + [M+H] + , 375.1151; the measured value is 375.1164.

[0219] (3) Synthesis of Compounds E1-E3

[0220]

[0221] 50 mg of compound 13 (0.13 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 51 mg of HATU (0.13 mmol, 1.0 equiv.) and 52 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.13 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds E1-E3; the compounds E1-E3 are arranged in the following order:

[0222]

[0223] Example 31 Synthesis of Compound E1

[0224] The synthesis steps were as in Example 30. The amine reagent in the last step was 5-aminoindolin-2-one to obtain compound E1 with a yield of 33%. 1 H NMR (500 MHz, DMSO-d 6 )δ13.52(br, 1H), 12.09(s, 1H), 10.36(s, 1H), 7.98(s, 1H), 7.93(d, J=7.5Hz, 1H) , 7.87 (d, J=8.0Hz, 1H), 7.82 (s, 1H), 7.75 (d, J=8.0Hz, 1H), 7.57 (dd, J=8.0, 2.0Hz , 1H), 7.43-7.36 (m, 2H), 7.22 (d, J=74.0Hz, 1H), 6.83 (d, J=8.5Hz, 1H), 4.07 (d, J= 7.0Hz, 2H), 3.52(s, 2H), 1.40-1.31(m, 1H), 0.67-0.60(m, 2H), 0.45-0.39(m, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ176.3, 162.4, 151.2, 150.1, 141.6, 140.7, 139.7, 135.3, 133.2, 126.8, 126.5, 122.9, 122.8, 122.0, 121.3, 119.5, 118.5, 116.6 (t, J = 256.5 Hz), 116.5, 115.4, 112.5, 109.2, 73.1, 36.2, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated for C 27 H 22 O 4 N 4 F 2 Na + [M+Na] + , 527.1501; the measured value is 527.1534.

[0225] Example 32 Synthesis of Compound E2

[0226] The synthesis steps were as in Example 30. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound E2 with a yield of 51%. 1 H NMR (500 MHz, DMSO-d 6 )δ13.56 (s, 1H), 12.12 (s, 1H), 10.11 (s, 1H), 8.02 (s, 1H), 7.96 (d, J=8.0Hz, 1H), 7.90 (dd, J =8.0, 2.0Hz, 1H), 7.79 (d, J = 8.0Hz, 1H), 7.75 (d, J = 2.0Hz, 1H), 7.60 (dd, J = 8.5, 2.0Hz, 1H), 7 .47-7.39 (m, 2H), 7.23 (t, J=74.0Hz, 1H), 6.91 (d, J=8.5Hz, 1H), 4.11 (d, J=2.0Hz, 2H), 2.94 ( t, J=7.5Hz, 2H), 2.50-2.45(m, 2H), 1.42-1.30(m, 1H), 0.68-0.61(m, 2H), 0.45-0.39(m, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ170.0, 162.4, 151.3, 150.1, 141.6, 140.7, 135.3, 134.4, 133.5, 126.8, 124.3, 123.0, 122.9, 121.9, 121.4, 119.5, 119.0, 118.2, 116.6 (d, J = 256.5 Hz), 115.5, 115.4, 112.5, 73.1, 30.4, 25.2, 10.0, 3.2 (2×C). ESI-HRMS m / z: calculated for C 28 H 24 O 4 N 4 F 2 Na + [M+Na] + , 541.1658; the measured value is 541.1680.

[0227] Example 33 Synthesis of Compound E3

[0228] Synthesis steps refer to Example 30, the amine reagent in the last step of the reaction is 6-(4-aminophenyl)-3(2H)-pyridazinone to obtain compound E3 with a yield of 45%; Conformer A: 1 H NMR (500 MHz, DMSO-d 6 )δ13.25(s, 1H), 13.14(s, 1H), 10.47(s, 1H), 8.27(d, J=123.5Hz, 1H), 8.0 5 (s, 1H), 8.00-7.92 (m, 3H), 7.92-7.84 (m, 3H), 7.83 (s, 1H), 7.73 (d, J=61. 0Hz, 1H), 7.39 (d, J=3.5Hz, 1H), 7.21 (t, J=74.0Hz, 1H), 7.00 (s, 1H), 4.05 (d, J=7.0Hz, 2H), 1.37-1.27(m, 1H), 0.66-0.59(m, 2H), 0.46-0.38(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.1, 160.2, 152.3, 150.1, 146.2, 143.6, 141.3, 140.5, 137.5, 131.3 (2×C), 130.1, 129.5, 128.6, 127.9, 126. 0(2×C), 122.9, 121.3, 120.2, 119.3, 118.7, 116.6(t, J=256.5Hz), 112.4, 111.2, 73.3, 10.0, 3.1(2×C).Conformer B:1 H NMR (500 MHz, DMSO-d 6 )δ13.21(s, 1H), 13.14(s, 1H), 10.39(s, 1H), 8.27(d, J=123.5Hz, 1H), 8.0 3(s, 1H), 8.00-7.92(m, 3H), 7.92-7.84(m, 3H), 7.82(s, 1H), 7.72(d, J=61. 0Hz, 1H), 7.38 (d, J=3.5Hz, 1H), 7.21 (d, J=74.0Hz, 1H), 6.98 (s, 1H), 4.05 (d, J=7.0Hz, 2H), 1.37-1.27(m, 1H), 0.66-0.59(m, 2H), 0.46-0.38(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.0, 160.2, 152.9, 150.1, 146.2, 143.2, 141.4, 140.5, 134.7, 131.3 (2×C), 130.1, 129.5, 129.2, 127.8, 126.0 (2×C), 121.8, 121.3, 120.3, 119.3, 118.4, 116.6 (t, J=256.5 Hz), 112.5, 111.5, 73.3, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 29 H 23 O 4 N 5 F 2 Na + [M+Na] + , 566.1610; the measured value is 566.1601.

[0229] Example 34 Synthesis of F series compounds

[0230] (1) Synthesis of Compound 14

[0231]

[0232] 1.0 g of compound 1 (7.24 mmol, 1.0 equiv.) was dissolved in 10 mL of DMF solvent, and then 0.72 g of sodium hydroxide (18.10 mmol, 2.5 equiv.) and 2.52 g of ethyl difluorochloroacetate (15.93 mmol, 2.2 equiv.) were added, and the mixture was heated to 80° C. for 12 h and monitored by thin layer chromatography. After the reaction was completed, water was added to quench the reaction, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was dried over anhydrous sodium sulfate, and the organic phase was concentrated. 0.93 g of compound 14 was obtained by flash column chromatography with a yield of 54%.

[0233] The obtained compound 14 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (400 MHz, CDCl 3 )δ9.91(s,1H),7.75-7.73(m,2H),7.39(d,J=8.8Hz,1H),6.64(t,J=72.8Hz,1H),6.59(t,J=72.8Hz,1H).ESI-MS m / z: calculated value is C 9 H 7 O 3 F 4 + [M+H] + , 239.0; the measured value is 239.0.

[0234] (2) Synthesis of Compound 15

[0235]

[0236] The synthesis steps were similar to those of compound 10. Compound 14 was reacted with methyl 2,3-diaminobenzoate to obtain a yellow solid product with a yield of 50%.

[0237] The obtained compound 15 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (400 MHz, CDCl 3 )δ10.75 (br, 1H), 8.04 (d, J = 2.0 Hz, 1H), 8.01 (d, J = 8.0 Hz, 1H), 7.93-7.91 (m, 2H), 7.41 (d, J = 8.4 Hz, 1H), 7.32 (t, J = 8.0 Hz, 1H), 6.65 (t, J = 73.2 Hz, 1H), 6.62 (t, J = 72.8 Hz, 1H), 4.03 (s, 3H). ESI-HRMS m / z: calculated value is C 17 H 13 O 4 N 2 F4 + [M+H] + , 385.0806; the measured value is 385.0801.

[0238] (3) Synthesis of Compound 16

[0239]

[0240] The synthesis steps were similar to those of compound 5, and a white solid product was obtained with a yield of 90%.

[0241] The obtained compound 16 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (500 MHz, DMSO-d 6 )δ12.57 (br, 1H), 12.52 (br, 1H), 8.33 (s, 1H), 8.30 (d, J = 8.5 Hz, 1H), 7.95 (d, J = 8.0 Hz, 1H), 7.84 (dd, J = 7.5, 1.0 Hz, 1H), 7.52 (d, J = 8.5 Hz, 1H), 7.49-7.19 (m, 3H). ESI-HRMS m / z: calculated value is C 16 H 11 O 4 N 2 F 4 N + [M+H] + , 371.0649; the measured value is 371.0652.

[0242] (4) Synthesis of Compounds F1-F4

[0243]

[0244] 50 mg of compound 16 (0.13 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 51 mg of HATU (0.13 mmol, 1.0 equiv.) and 52 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.13 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and compounds F1-F4 were obtained by column chromatography; the compounds F1-F4 are as follows:

[0245]

[0246] Example 35 Synthesis of Compound F1

[0247] The synthesis steps were as in Example 34. The amine reagent in the last step was 6-aminoindolone to obtain compound F1 with a yield of 44%. 1 H NMR (600 MHz, DMSO-d 6 )δ13.71 (br, 1H), 12.18 (s, 1H), 10.46 (s, 1H), 8.25 (s, 1H), 8.22 (d, J = 9.0Hz, 1H), 7.98 (d, J = 7.2Hz, 1H), 7.83 (s, 1H), 7.81 (s, 1H), 7.63 (d, J =8.4Hz, 1H), 7.43 (t, J = 7.8Hz, 1H), 7.37 (t, J = 73.2Hz, 1H), 7.37 (d, J = 73.2Hz, 1H), 7.20 (d, J = 7.8Hz, 1H), 7.07 (d, J = 8.4Hz, 1H), 3.46 (s, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.8, 162.6, 150.3, 144.4, 143.5, 142.0, 140.7, 138.4, 135.4, 126.9, 124.9, 124.6, 123.2, 123.1, 122.0, 121.3, 120.8, 119.4, 116.6 (t, J = 259.5 Hz), 116.3 (t, J = 259.5 Hz), 115.9, 111.8, 101.1, 35.5. ESI-HRMS m / z: calculated value is C 24 H 16 O 4 N 4 F 4 Na + [M+Na] + , 523.1000; the measured value is 523.1016.

[0248] Example 36 Synthesis of Compound F2

[0249] The synthesis steps were as in Example 34. The amine reagent used in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound F2 with a yield of 23%. 1 H NMR (600 MHz, DMSO-d 6)δ14.11(s, 1H), 12.14(s, 1H), 10.20(s, 1H), 8.34(s, 1H), 8.31(d, J=8.4Hz, 1H) , 7.98 (d, J = 7.8Hz, 1H), 7.82 (d, J = 8.4Hz, 1H), 7.66 (s, 1H), 7.64 (d, J = 8.4Hz, 1H) , 7.45 (t, J=74.0Hz, 1H), 7.43 (d, J=4.8Hz, 1H), 7.39 (t, J=73.2Hz, 1H), 7.26 (d, J =7.2Hz, 1H), 7.18 (d, J = 8.4Hz, 1H), 2.87 (t, J = 7.8Hz, 2H), 2.47 (t, J = 7.8Hz, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ170.4, 162.5, 150.3, 143.4, 142.0, 140.7, 138.9, 137.9, 135.4, 128.1, 126.8, 125.1, 123.2, 123.1, 121.9, 121.3, 119.3, 119.0, 116.6 (t, J = 259.5 Hz), 116.4 (t, J = 259.5 Hz), 115.9, 112.8, 106.4, 30.6, 24.4. ESI-HRMS m / z: calculated for C 25 H 18 O 4 N 4 F 4 Na + [M+Na] + , 537.1156; the measured value is 537.1187.

[0250] Example 37 Synthesis of Compound F3

[0251] The synthesis steps were as in Example 34. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound F3 with a yield of 23%. 1 H NMR (500 MHz, DMSO-d 6)δ13.69 (s, 1H), 12.06 (s, 1H), 10.09 (s, 1H), 8.26 (d, J=2.0Hz, 1H), 8.20 (dd, J=8.5, 2.0Hz, 1H), 7.97 (d, J=7.5Hz, 1H), 7.80 (d, J=8.0Hz, 1H), 7.6 9(d, J=2.5Hz, 1H), 7.67-7.61(m, 2H), 7.58-7.26(m, 2H), 7.37(t, J=74.0 Hz, 1H), 6.90 (d, J=8.5Hz, 1H), 2.93 (t, J=7.0Hz, 2H), 2.49-2.44 (m, 2H). 13 CNMR (151 MHz, DMSO-d 6 )δ169.9, 162.3, 150.2, 143.4, 142.0, 140.6, 135.3, 134.4, 133.4, 126.8, 124.8, 124.2, 123.2, 123.1, 122.1, 121.4, 119.1, 118.9, 118.1, 116.6 (t, J = 259.5 Hz), 116.3 (t, J = 259.5 Hz), 115.7, 115.4, 30.4, 25.0. ESI-HRMS m / z: calculated for C 25 H 18 O 4 N 4 F 4 Na + [M+Na] + , 537.1156; the measured value is 537.1185.

[0252] Example 38 Synthesis of Compound F4

[0253] The synthesis steps were as in Example 34. The amine reagent used in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound F4 with a yield of 44%. 1 H NMR (500 MHz, DMSO-d 6) δ13.75 (s, 1H), 12.34 (s, 1H), 10.94 (s, 1H), 8.28 (d, J = 2.0Hz, 1H), 8.23 ​​(dd, J = 9.0, 2. 0Hz, 1H), 8.00 (d, J=7.5Hz, 1H), 7.94 (d, J=8.5Hz, 2H), 7.88-7.81 (m, 3H), 7.64 (d, J=8. 5Hz, 1H), 7.45 (t, J=7.5Hz, 1H), 7.42 (d, J=74.0Hz, 1H), 7.37 (d, J=74.0Hz, 1H), 3.48-3 .39 (m, 1H), 2.72 (dd, J=17.0, 7.0Hz, 1H), 2.26 (d, J=16.5Hz, 1H), 1.11 (d, J=7.5Hz, 3H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.3, 162.8, 152.1, 150.3, 143.5, 142.0, 140.7, 139.9, 135.3, 130.0, 126.7, 126.6 (2×C), 124.9, 123.3, 123.2, 121.8, 121.4, 119.3, 119.1 (2×C), 116.6 (t, J=258.0 Hz), 116.3 (t, J=259.5 Hz), 116.0, 33.5, 27.0, 15.9. ESI-HRMS m / z: calculated value is C 27 H 21 O 4 N 5 F 4 Na + [M+Na] + , 578.1422; the measured value is 578.1450.

[0254] Example 39 Synthesis of G series compounds

[0255] (1) Synthesis of Compound 18

[0256]

[0257] The synthesis steps refer to the synthesis of compound 2. 3,4-Dihydroxyacetophenone and ethyl difluorochloroacetate are reacted to obtain a white solid product with a yield of 36%.

[0258] The obtained compound 18 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (600 MHz, CDCl 3)δ7.63 (d, J=1.8 Hz, 1H), 7.51 (dd, J=9.0, 2.4 Hz, 1H), 7.18 (d, J=8.4 Hz, 1H), 6.64 (t, J=73.2 Hz, 1H), 6.33 (br, 1H), 2.58 (s, 3H). ESI-HRMS m / z: calculated value is C 9 H 8 O 3 F 2 Na + [M+Na] + , 225.0334; the measured value is 225.0311.

[0259] (2) Synthesis of Compound 19

[0260]

[0261] The synthesis steps were similar to those of compound 3, and a yellow oily product was obtained with a yield of 56%.

[0262] The obtained compound 19 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, DMSO-d 6 )δ7.87 (dd, J=8.4, 2.0 Hz, 1H), 7.69 (d, J=2.0 Hz, 1H), 7.24 (d, J=8.8 Hz, 1H), 7.17 (t, J=74.4 Hz, 1H), 3.99 (d, J=7.2 Hz, 2H), 2.53 (s, 3H), 1.30-1.23 (m, 1H), 0.65-0.52 (m, 2H), 0.43-0.27 (m, 2H). ESI-HRMS m / z: calculated value is C 13 H 14 O 3 F 2 Na + [M+Na] + , 279.0803; the measured value is 279.0808.

[0263] (3) Synthesis of Compound 20

[0264]

[0265] 2 g of compound 19 (7.81 mmol, 1.0 equiv.) and 1.13 g of N, N-dimethylformamide dimethyl acetal (9.37 mmol, 1.2 equiv.) were weighed into a reaction bottle, and 2 mL of N, N-dimethylformamide was added. The mixture was stirred overnight at 120°C and monitored by TLC. The mixture was extracted with ethyl acetate three times, the organic phase was washed once with a saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was concentrated and subjected to flash column chromatography to obtain 1.8 g of a yellow solid product with a yield of 74%.

[0266] The obtained compound 20 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, DMSO-d 6 )δ7.70 (d, J = 12.4 Hz, 1H), 7.57-7.47 (m, 2H), 7.19 (d, J = 8.0 Hz, 1H), 7.16 (t, J = 74.4 Hz, 1H), 5.81 (d, J = 12.4 Hz, 1H), 3.94 (d, J = 6.8 Hz, 2H), 3.14 (s, 3H), 2.92 (s, 3H), 1.27 (d, J = 8.1 Hz, 1H), 0.61-0.49 (m, 2H), 0.42-0.30 (m, 2H). ESI-HRMS m / z: calculated value is C 16 H 19 O 3 NF 2 Na + [M+Na] + , 334.1225; the measured value is 334.1225.

[0267] (4) Synthesis of Compound 21

[0268]

[0269] 2.48 g of compound 20 (7.97 mmol, 1.0 equiv.) and 1.46 g of 5-amino-1H-pyrazole-3-carboxylic acid methyl ester (23.91 mmol, 3.0 equiv.) were weighed into a reaction bottle, 10 mL of acetic acid was added, and the mixture was stirred at 80°C overnight and monitored by TLC. The mixture was extracted with ethyl acetate three times, the organic phase was washed once with a saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate. The organic phase was concentrated and subjected to rapid column chromatography to obtain 2.6 g of a white solid product with a yield of 84%.

[0270] The obtained compound 21 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (400 MHz, DMSO-d 6)δ8.72 (d, J=4.4 Hz, 1H), 7.89 (d, J=2.0 Hz, 1H), 7.71 (dd, J=8.4, 2.0 Hz, 1H), 7.45-7.39 (m, 2H), 7.28 (s, 1H), 7.27 (t, J=74.0 Hz, 1H), 3.99 (d, J=6.8 Hz, 2H), 3.88 (s, 3H), 1.42-1.26 (m, 1H), 0.66-0.51 (m, 2H), 0.43-0.24 (m, 2H). ESI-HRMS m / z: calculated value is C 19 H 17 O 4 N 3 F 2 Na + [M+Na] + , 412.1079; the measured value is 412.1083.

[0271] (51 Synthesis of Compound 22

[0272]

[0273] The synthesis steps were similar to those of compound 5, and a yellow solid product was obtained with a yield of 98%.

[0274] The obtained compound 22 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400 MHz, DMSO-d 6 )δ8.58 (d, J = 4.4 Hz, 1H), 7.85 (d, J = 1.6 Hz, 1H), 7.78 (dd, J = 8.4, 2.0 Hz, 1H), 7.38 (d, J = 8.4 Hz, 1H), 7.27 (d, J = 4.0 Hz, 1H), 7.25 (t, J = 74.0 Hz, 1H), 6.95 (s, 1H), 3.99 (d, J = 6.8 Hz, 2H), 1.37-1.30 (m, 1H), 0.65-0.54 (m, 2H), 0.39-0.26 (m, 2H), OH (not observed). ESI-HRMS m / z: calculated for C 18 H 15 O 4 N 3 F 2 Na + [M+Na] + , 398.0923; the measured value is 398.0909.

[0275] (6) Synthesis of Compounds G1-G4

[0276]

[0277] 50 mg of compound 22 (0.13 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 51 mg of HATU (0.13 mmol, 1.0 equiv.) and 52 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.13 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds G1-G4; the compounds G1-G4 are as follows:

[0278]

[0279] Example 40 Synthesis of Compound G1

[0280] Synthesis steps: Refer to Example 39, the amine reagent in the last step of the reaction is 5-aminodihydroindole-2-one, and compound G1 is obtained with a yield of 38%; Conformer A: 1 H NMR (600 MHz, DMSO-d 6 )δ10.20 (s, 1H), 10.16 (s, 1H), 8.56 (d, J = 4.2Hz, 1H), 7.93 (s, 1H), 7.75 (d, J = 8.4Hz, 1H), 7.69 (d, J=4.2Hz, 1H), 7.59 (s, 1H), 7.48 (d, J=8.4Hz, 1H), 7.27 (d, J=8.4Hz, lH), 7.14 (s, 1H), 7.13 (t, J=74.4Hz, 1H), 6.68 (s, 1H), 4.06 (d, J=3.0 Hz, 2H), 3.50 (s, 2H), 1.38-1.32 (m, 1H), 0.63-0.58 (m, 2H), 0.43-0.37 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.4, 159.6, 155.3, 150.3, 150.1, 148.2, 144.8, 142.0, 140.1, 136.4, 132.4, 122.6, 126.1, 120.6 , 120.0, 117.7, 116.6 (t, J=256Hz), 115.9, 108.8, 107.6, 96.9, 73.4, 36.1, 10.0, 3.1 (2×C).Conformer B: 1HNMR (600 MHz, DMSO-d 6 )δ10.22(s, 1H), 9.93(s, 1H), 9.06(d, J=7.2Hz, 1H), 7.76(s, 1H), 7.71(d, J=8 .4Hz, 1H), 7.32 (d, J=4.2Hz, 1H), 7.53 (s, 1H), 7.41 (d, J=8.4Hz, 1H), 7.20 (d, J =8.4Hz, 1H), 7.05 (s, 1H), 7.08 (t, J = 74.4Hz, 1H), 6.65 (s, 1H), 4.05 (d, J = 3.0 Hz, 2H), 3.50 (s, 2H), 1.32-1.26 (m, 1H), 0.59-0.52 (m, 2H), 0.37-0.30 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.3, 159.4, 151.2, 150.6, 150.0, 149.4, 144.8, 141.9, 140.0, 134.3, 132.5, 127.9, 126.0, 120.9, 120.3, 117.7, 108.8, 116.5 (t, J = 256 Hz), 112.9, 109.4, 97.0, 73.3, 36.0, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 26 H 21 O 4 N 5 F 2 Na + [M+Na] + , 528.1454; the measured value is 528.1469.

[0281] Example 41 Synthesis of Compound G2

[0282] The synthesis steps were as in Example 39. The amine reagent in the last step was 6-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound G2 with a yield of 20%. 1 H NMR (600 MHz, DMSO-d 6) δ10.09 (s, 1H), 10.07 (s, 1H), 8.71 (d, J = 4.2Hz, 1H), 8.09 (s, 1H), 7.88 (d, J = 9.0Hz, 1H), 7.61 (s, 1H), 7.55 (dd, J=8.4, 2.4Hz, 1H), 7.47 (d, J=4.2Hz, 1H), 7.42 (d, J=8.4Hz, 1H), 7.2 8 (s, 1H), 7.28 (t, J=74.4Hz, 1H), 6.84 (d, J=8.4Hz, 1H), 4.06 (d, J=6.6Hz, 2H), 2.88 (t, J=7 .2Hz, 2H), 2.46 (t, J=7.2Hz, 2H), 1.38-1.31 (m, 1H), 0.59-0.52 (m, 2H), 0.37-0.30 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ170.0, 159.5, 155.4, 151.1, 150.1, 148.2, 142.1, 136.4, 134.6, 134.3, 132.8, 123.7, 120.9, 120.3, 120.2, 119.5, 116.6 (t, J = 256.5 Hz), 115.0, 112.9, 107.7, 96.9, 73.3, 30.4, 25.1, 10.0, 3.1 (2×C). ESI-HRMS m / z: calculated for C 27 H 23 O 4 N 5 F 2 Na + [M+Na] + , 542.1610; the measured value is 542.1624.

[0283] Example 42 Synthesis of Compound G3

[0284] Synthesis steps: Refer to Example 39, the amine reagent in the last step of the reaction is 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound G3 with a yield of 12%; 1 H NMR (500 MHz, DMSO-d 6)δ10.93 (s, 1H), 10.73 (s, 1H), 9.25 (d, J = 7.5Hz, 1H), 7.99 (s, 1H), 7.97 (s, 1H), 7.93 (d, J = 2.0Hz, 1H ), 7.89 (d, J = 7.5Hz, 1H), 7.87 (dd, J = 8.5, 2.0Hz, 1H), 7.80 (s, 1H), 7.79 (s, 1H), 7.36 (d, J = 8.5Hz, 1H) , 7.34 (s, 1H), 7.25 (d, J=74.0Hz, 1H), 4.06 (d, J=7.0Hz, 2H), 3.44-3.38 (m, 1H), 2.74-2.65 (m, 1H), 2. 24 (d, J=16.5Hz, 1H), 1.34-1.25 (m, 1H), 1.09 (d, J=7.5Hz, 3H), 0.64-0.57 (m, 2H), 0.43-0.37 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ166.4, 160.2, 152.2, 150.8, 150.1, 150.0, 149.4, 144.9, 141.9, 139.5, 130.3, 127.9, 126.3 (2×C), 122.6, 120.6, 120.2 (2×C), 116.6 (t, J=257.0 Hz), 115.9, 109.6, 97.3, 73.4, 33.6, 27.0, 16.0, 9.9, 3.2 (2×C). ESI-HRMS m / z: calculated value is C 29 H 26 O 4 N 6 F 2 Na + [M+Na] + , 583.1876; the measured value is 583.1894.

[0285] Example 43 Synthesis of Compound G4

[0286] The synthesis steps were as in Example 39. The amine reagent in the last step was 6-amino-3(2H)-pyridazinone to obtain compound G4 with a yield of 30%. 1 H NMR (500 MHz, DMSO-d 6)δ12.86 (s, 1H), 10.60 (s, 1H), 8.73 (d, J = 4.5Hz, 1H), 7.96 (d, J = 10.0Hz, 1H), 7.95-7.91 (m, 2H), 7.48 (d, J = 4.5Hz, 1H), 7.41 (d, J = 8.0Hz, 1H) , 7.38 (s, 1H), 7.27 (t, J=74.5Hz, 1H), 6.98 (d, J=10.0Hz, 1H), 4.03 (d, J=7.0Hz, 2H), 1.38-1.28(m, 1H), 0.60-0.54(m, 2H), 0.38-0.31(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ160.6, 160.0, 150.8, 149.9, 149.5, 148.7, 145.0, 141.9, 140.7, 130.8, 130.4, 127.8, 122.7, 120.5, 116.5 (d, J = 256.5 Hz), 115.8, 109.8, 97.5, 73.4, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 22 H 18 O 4 N 6 F 2 Na + [M+Na] + , 491.1250; the measured value is 491.1266.

[0287] Example 44 Synthesis of H series compounds

[0288] (1) Synthesis of Compound 23

[0289]

[0290] The synthesis steps were similar to those of compound 21. Compound 20 was reacted with methyl 3-aminopyrazole-4-carboxylate to obtain a yellow solid product with a yield of 56%.

[0291] The obtained compound 23 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (500 MHz, CDCl 3)δ8.79(d, J=4.5 Hz, 1H), 8.61(s, 1H), 7.70(d, J=2.0 Hz, 1H), 7.51(dd, J=8.5, 2.0 Hz, 1H), 7.33(d, J=8.5 Hz, 1H), 7.07(d, J=4.5 Hz, 1H), 6.74(t, J=74.5 Hz, 1H), 3.97(s, 3H), 3.94(d, J=7.0 Hz, 2H), 1.39-1.27(m, 1H), 0.76-0.52(m, 2H), 0.41-0.32(m, 2H).ESI-HRMS m / z: calculated value is C 19 H 17 O 4 N 3 F 2 Na + [M+Na] + , 412.1079; the measured value is 412.1072.

[0292] (2) Synthesis of Compound 24

[0293]

[0294] The synthesis steps were similar to those of compound 5, and a yellow solid product was obtained with a yield of 98%.

[0295] The obtained compound 24 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result was: 1 H NMR (600 MHz, DMSO-d 6 )δ8.62(d, J=4.2 Hz, 1H), 8.33(s, 1H), 7.80(d, J=1.8 Hz, 1H), 7.74(dd, J=8.4,1.8 Hz, 1H), 7.38(d, J=8.4 Hz, 1H), 7.24(d, J=4.2 Hz, 1H), 7.23(t, J=73.8 Hz, 1H), 3.98(d, J=7.2 Hz, 2H), 1.35-1.24(m, 1H), 0.64-0.54(m, 2H), 0.40-0.30(m, 2H).OH(not observed).ESI-HRMS m / z: calculated for C 18 H 15 O 4 N 3 F 2 Na + [M+Na] + , 398.0923; the measured value is 398.0915.

[0296] (3) Synthesis of compounds H1-H4

[0297]

[0298] 50 mg of compound 24 (0.13 mmol, 1.0 equiv.) was weighed and dissolved in N, N-dimethylformamide, 51 mg of HATU (0.13 mmol, 1.0 equiv.) and 52 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added, stirred at room temperature for 20 min, and the corresponding amine reagent (0.13 mmol, 1.0 equiv.) was added, and the reaction was carried out at room temperature for 6 h. Monitored by TLC, water was added to quench the reaction after the raw material reaction was complete, extracted with ethyl acetate 3 times, washed once with saturated sodium chloride solution, the ethyl acetate layer was dried with anhydrous sodium sulfate, the solution was concentrated, and purified by column chromatography to obtain compounds H1-H4; the compounds H1-H4 are as follows:

[0299]

[0300] Example 45 Synthesis of Compound H1

[0301] The synthesis steps were as in Example 44. The amine reagent in the last step was 6-aminoindolone to obtain compound H1 with a yield of 78%. 1 H NMR (600 MHz, DMSO-d 6 )δ10.43 (s, 1H), 10.10 (s, 1H), 8.94 (d, J = 4.8Hz, 1H), 8.73 (s, 1H), 7.86 (d, J = 1.8Hz, 1H) , 7.81 (dd, J=8.4, 1.8Hz, 1H), 7.58 (d, J=4.8Hz, 1H), 7.52 (d, J=1.8Hz, 1H), 7.42 (d, J=8. 4Hz, 1H), 7.27 (t, J=74.4Hz, 1H), 7.17 (d, J=7.8Hz, 1H), 7.09 (dd, J=7.8, 1.8Hz, 1H), 4.0 0(d, J=7.2Hz, 2H), 3.43(s, 2H), 1.34-1.27(m, 1H), 0.63-0.58(m, 2H), 0.40-0.35(m, 2H). 13 C NMR (151 MHz, DMSO-d 6)δ176.8, 159.4, 152.1, 149.5, 146.5, 146.5, 145.8, 144.3, 142.2, 138.1, 127.5, 124.6, 122.9, 120.6, 120.5, 116.5 (d, J = 256.5 Hz), 116.0, 111.7, 109.6, 105.1, 100.9, 73.4, 35.4, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated for C 26 H 21 O 4 N 5 F 2 Na + [M+Na] + , 528.1454; the measured value is 528.1454.

[0302] Example 46 Synthesis of Compound H2

[0303] The synthesis steps were as in Example 44. The amine reagent in the last step was 5-aminoindolin-2-one to obtain compound H2 with a yield of 77%. 1 H NMR (500 MHz, DMSO-d 6 )δ10.35 (s, 1H), 9.99 (s, 1H), 8.93 (d, J = 4.5Hz, 1H), 8.72 (s, 1H), 7.86 (s, 1H), 7 .81 (d, J=8.5Hz, 1H), 7.66 (s, 1H), 7.57 (d, J=4.5Hz, 1H), 7.49 (d, J=8.5Hz, 1H), 7 .43 (d, J=8.5Hz, 1H), 7.19 (t, J=74.0Hz, 1H), 6.82 (d, J=8.5Hz, 1H), 4.00 (d, J=6 .5Hz, 2H), 3.51(s, 2H), 1.37-1.27(m, 1H), 0.68-0.54(m, 2H), 0.45-0.32(m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.3, 159.2, 152.1, 149.5, 146.5, 146.4, 145.7, 142.1, 139.7, 132.7, 127.6, 126.4, 122.8, 120.5, 118.8, 116.8, 116.5 (t, J = 256.5 Hz), 116.0, 109.5, 109.1, 105.2, 73.4, 36.1, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated for C 26 H 21 O4 N 5 F 2 Na + [M+Na] + , 528.1454; the measured value is 528.1470.

[0304] Example 47 Synthesis of Compound H3

[0305] The synthesis steps were as in Example 44. The amine reagent used in the last step was 7-amino-3,4-dihydro-2(1H)-quinolinone to obtain compound H3 with a yield of 57%. 1 H NMR (500 MHz, DMSO-d 6 )δ10.12 (s, 1H), 10.02 (s, 1H), 8.94 (d, J = 3.6Hz, 1H), 8.74 (s, 1H), 7.87 (d, J = 1.6Hz, 1H), 7.81 (dd, J=6.8, 1.6Hz, 1H), 7.58 (d, J=3.6Hz, 1H), 7.45-7.40 (m, 2H), 7.27 (t, J=59.2Hz, 1 H), 7.20 (dd, J=6.4, 1.6Hz, 1H), 7.15 (d, J=6.4Hz, 1H), 4.01 (d, J=6.0Hz, 2H), 2.85 (t, J=6 .8Hz, 2H), 2.45 (t, J=6.4Hz, 2H), 1.36-1.30 (m, 1H), 0.63-0.57 (m, 2H), 0.39-0.35 (m, 2H). 13 C NMR (101 MHz, DMSO-d 6 )δ170.3, 159.4, 152.2, 149.5, 146.6, 146.5, 145.9, 142.2, 138.8, 137.6, 128.1, 127.6, 122.9, 120.5, 118.8, 116.5, 116.0 (t, J = 257.4 Hz), 113.0, 109.6, 106.4, 105.1, 73.4, 30.6, 24.4, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated for C 27 H 23 O 4 N 5 F 2 Na + [M+Na] + , 542.1610; the measured value is 542.1633.

[0306] Example 48 Synthesis of Compound H4

[0307] The synthesis steps were as in Example 44. The amine reagent used in the last step was 6-(4-aminophenyl)-4,5-dihydro-5-methyl-3(2H)-pyridazinone to obtain compound H4 with a yield of 36%. 1 H NMR (500 MHz, DMSO-d 6 ) δ10.92 (s, 1H), 10.26 (s, 1H), 8.96 (d, J = 4.5Hz, 1H), 8.76 (s, 1H), 7.87 (d, J = 2.0Hz, 1H), 7. 82 (s, 4H), 7.79 (d, J=2.0Hz, 1H), 7.59 (d, J=4.5Hz, 1H), 7.43 (d, J=8.0Hz, 1H), 7.27 (t, J=74. 0Hz, 1H), 4.00 (d, J=7.0Hz, 2H), 3.42-3.39 (m, 1H), 2.70 (dd, J=16.5, 7.0Hz, 1H), 2.25 (d, J= 16.5Hz, 1H), 1.34-1.28 (m, 1H), 1.09 (d, J=7.5Hz, 3H), 0.63-0.58 (m, 2H), 0.41-0.35 (m, 2H). 13 C NMR (151 MHz, DMSO-d 6 )δ166.4, 159.6, 152.3, 152.2, 149.6, 146.7 (2×C), 145.9, 142.2, 139.7, 129.8, 127.6, 126.6 (2×C), 122.9, 120.6, 119.2 (2×C), 116.5 (d, J=256.0 Hz), 116.0, 109.8, 105.0, 73.5, 33.6, 27.0, 16.0, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated value is C 29 H 26 O 4 N 6 F 2 Na + [M+Na] + , 583.1876; the measured value is 583.1889.

[0308] Active Examples

[0309] Activity Example 1 Detection of the Inhibition Rate of the Compounds of the Present Application on PDE3A and PDE4B1

[0310] 1. Preparation of reaction buffer and reaction stop solution (reagents see Table 1)

[0311] (1) Preparation of 1x reaction buffer

[0312] IMAP reaction buffer (provided by IMAP FP IPP Explorer Kit) containing 0.1% BSA (5×) was diluted into 1× reaction buffer containing 1 mM DTT.

[0313] (2) Preparation of reaction termination solution

[0314] The reaction termination solution was prepared by mixing IMAP Progressive Binding Buffer A (5×), IMAP Progressive Binding Buffer B (5×), and IMAP Progressive Binding Reagent (provided by IMAP FP IPP Explorer Kit) according to the instruction manual.

[0315] 2. Compound Preparation

[0316] (1) Compound dilution

[0317] Prepare a solution with a concentration of 100 times the final concentration of the compound to be tested. Use an automatic microporous pipette (PrecisionPRC384U) to dilute the compound gradient to the set number of concentration points, as follows: For a 5-fold dilution, add 50 μL of the compound DMSO solution at the starting concentration to well A2 on the Echo 384-well plate, and add 40 μL of 100% DMSO to wells A3-A11; take 10 μL of the compound from well A2 and add well A3, mix well, and make 5-fold dilutions in turn, diluting 10 concentration points; add 40 μL of 100% DMSO to wells A1 and A12.

[0318] (2) Transfer compounds to 384 reaction plates

[0319] Use the Echo550 instrument to transfer 200 nL of the diluted compound from the Echo384-well plate to a 384-well reaction plate, and transfer 200 nL of 100% DMSO to both the negative control and the positive control.

[0320] 3. Enzymatic reaction

[0321] (1) Prepare 2x enzyme solution

[0322] PDE4B1 was added to 1x reaction buffer to form a 2x enzyme solution (final concentration of PDE4B1: 0.00625 μg / ml).

[0323] (2) Prepare 2 times the substrate solution

[0324] For the enzyme PDE4B1, FAM-labeled cAMP was added to 1x reaction buffer to form a 2x substrate solution (final concentration of FAM-cAMP: 0.1 μM).

[0325] (3) Add enzyme solution to the 384-well plate

[0326] Add 10 μL of 2x enzyme solution to each well of a 384-well reaction plate. For the no-enzyme control well, replace the enzyme solution with 10 μL of 1x reaction buffer. Centrifuge at 1000 rpm for 1 min and incubate at room temperature for 15 min.

[0327] (4) Add substrate solution to the 384-well plate to start the enzymatic reaction

[0328] Add 10 μL of 2x substrate solution to each well of the 384-well reaction plate. Centrifuge at 1000 rpm for 1 min. The reaction was continued for 30 minutes.

[0329] (5) Termination of enzyme reaction

[0330] 60 μL of the reaction stop solution was added to each well of the 384-well reaction plate to terminate the reaction, and the plate was incubated at room temperature for 60 minutes on a shaker at 600 rpm in the dark.

[0331] 4. Read data and calculate data using EnVision

[0332] Readings were taken with EnVision.

[0333] 5. Calculation of Inhibition Rate and IC 50 Curve Fitting

[0334] Data were copied from EnVision, where the maximum value refers to the reading of the DMSO control and the minimum value refers to the reading of the no enzyme activity control. Inhibition rate (%) = (maximum value - sample value) / (maximum value - minimum value) × 100%.

[0335] The data were imported into MS Excel and IC fitted using XLFit excel add-in version 5.4.0.8 50 value;

[0336] Fitting formula: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope)

[0337] For the detection method of PDE3A, refer to PDE4B1.

[0338] Table 1 Reagent information

[0339] name brand Part Number PDE4B1 BPS 60041 PDE3A BPS 60032 Trequinsin TOCRIS 2337 / 10 IMAP FP IPP Kit Molecular Device R8124 FAM-CAMP Molecular Device R7506

[0340] The inhibitory effects of the compounds of the examples of the present application on PDE3A and PDE4B1 enzymes were detected according to the above method, and the results are shown in Table 2. The results of the determination of the inhibitory effects of the compounds of the examples of the present application on PDE3A and PDE4B1 enzymes are listed in Table 2.

[0341] Table 2 Inhibition rate of compounds on PDE3A and PDE4B1

[0342]

[0343] IC of the compound of Activity Example 2 for PDE3A and PDE4B1 50 Detection

[0344] Implementation steps refer to Activity Example 1 to obtain IC 50 value.

[0345] IC of compounds B5, C5, D5, D6, D7, F4 and G3 50 The values ​​are shown in Table 3.

[0346] Table 3 Compound IC 50 value

[0347] Compound <![CDATA[PDE3A IC 50 (nM)]]> <![CDATA[PDE4B1 IC 50 (nM)]]> B5 44 ＞500 C5 163 395 D5 10 9.4 D6 97 5.3 D7 1370 14 F4 78 1.7 G3 20 57

[0348] In vitro anti-inflammatory activity of the compound of Activity Example 3

[0349] The test method adopts the conventional ELISA method to carry out the inhibition test of the expression level of cellular inflammatory factors on the compounds of the present application.

[0350] 1. Cell Culture

[0351] Mouse mononuclear macrophage Raw 264.7 cells were cultured in DMEM high-glucose medium containing 10% (V / V) FBS, 100 μg / mL penicillin, and 100 μg / mL streptomycin. The culture conditions were 37°C and 5% CO. 2 The cells were passaged when they reached 80% confluence.

[0352] 2. Cellular administration and induction of cellular inflammation

[0353] The experiment was conducted using cells in the logarithmic growth phase. The cells were cultured at a rate of 2×10 5 / well of a 12-well plate and incubate at 37°C, 5% CO 2Culture in an environment until the cells grow to 70% and set aside. Set up groups: blank group, LPS stimulation group, positive control group (dexamethasone) and drug-treated group (compound of the present application). Carefully remove the culture medium, add fresh complete culture medium containing the compound to the positive control group and the drug-treated group, respectively, and add equal volumes of DMSO to the blank group and LPS stimulation group. After 1 hour, 1 μg / mL LPS was added to all groups except the blank group, and cell inflammation was induced after 4 hours. Take out the well plate and perform the ELISA experiment according to the instructions of the kit.

[0354] Collect the culture medium supernatant: collect the supernatant into a 1.5 mL EP tube, centrifuge at 1000 rpm for 10 minutes, and take the supernatant for ELISA detection of cytokines.

[0355] The secretion of mouse IL-1β and IL-6 in the culture medium supernatant was detected using an enzyme-linked immunosorbent assay (ELISA) kit. The specific steps are as follows:

[0356] (1) Reagent preparation

[0357] ① Take out the refrigerator and place at room temperature to equilibrate for 20 minutes. ② Dilute the washing solution (20×) with double distilled water to 1× to prepare the required washing solution. ③ Add the standard diluent to 1 bottle of standard according to the volume marked on the standard label and incubate at room temperature for 15 minutes. ④ Take 5 clean 1.5 ml centrifuge tubes, add 250μL of standard diluent to each tube in advance, and dilute the standard in multiples to obtain a total of six standard concentrations of 1000, 500, 250, 125, 62.5, and 31.25pg / mL. Finally, add the diluted standards to the pre-coated plate wells in turn, and add the standard diluent directly as 0pg / mL concentration. There are a total of seven standard concentrations. ⑤ Add 300μL to each well, and proceed to the next time after about 15-30 seconds. Wash the plate five times in total and pat dry on paper.

[0358] (2) Operation steps

[0359] ① Calculate the number of pre-coated strips required for one experiment, take out the required strips and place them in the 96-well frame.

[0360] ② Add samples or standards of different concentrations into the corresponding wells at 100 μL / well, seal the reaction wells with a sealing film (transparent), and incubate at room temperature for 120 minutes.

[0361] ③ Wash the plate 5 times and pat dry on thick absorbent paper for the last wash.

[0362] ④ Add 100 μL / well of biotinylated antibody, seal the reaction wells with a sealing film (transparent), and incubate at room temperature for 60 minutes.

[0363] ⑤ Wash the plate 5 times and pat dry on thick absorbent paper for the last wash.

[0364] ⑥Add 100 μL / well of horseradish peroxidase-labeled Streptavidin, seal the reaction wells with a sealing film (white), and incubate at room temperature in the dark for 20 minutes.

[0365] ⑦ Wash the plate 5 times and pat dry on thick absorbent paper for the last wash.

[0366] ⑧Add 100 μL / well of TMB solution, seal the reaction wells with a sealing film (white), and incubate at room temperature in the dark for 20 minutes.

[0367] ⑨Add 50 μL / well of stop solution, mix well and measure the A450 value immediately.

[0368] The test results are as follows Figure 1 and Figure 2 shown.

[0369] Depend on Figure 1 It can be seen that the inhibitory effects of compounds D4, D1, A6, A2, A3 and A1 on the inflammatory factor IL-1β were higher than those of the control group. Figure 2 It can be seen that compounds A6, C5, D4, H5, E3 and D5 have good inhibitory effects on the inflammatory factor IL-6.

[0370] From the above, it can be seen that the compounds of the present application have good anti-inflammatory effects.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated form thereof in R is Ring A is a five-membered to ten-membered heteroaryl group, comprising 1-3 heteroatoms selected from N, O, and S, wherein at least one heteroatom is N; optionally, the five-membered to ten-membered heteroaryl group is substituted by one or more substituents selected from C1-C6 alkyl and C1-C6 alkoxy; Ring B is X1 and Y1 are each independently N or NH, O or S, and at least one of X1 and Y1 is N or NH; R1, R2, and R3 are each independently H, C1-C6 alkyl, or C1-C6 alkoxy; n is 1 or 2; *Indicates the connection location.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated product thereof, wherein the five-membered to ten-membered heteroaryl group is a monocyclic heteroaryl group or a condensed-ring heteroaryl group; Preferably, Ring A is X2, Y2, and Z2 are each independently N or NH, O or S, and at least one of X2, Y2, and Z2 is N or NH; Preferably, Ring A is 3. The compound according to claim 2, or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated substance thereof, wherein X2 is each independently N or NH, and Y2 and Z2 are each independently N or NH, O or S.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated form thereof, wherein Ring B is X1, Y1 are N or NH; R2 is H or C1-C6 alkyl; preferably, ring B is 5. The compound of claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated form thereof, wherein Formula I is R5 is Preferably, Formula I is R6 is Preferably, Formula I is R7 is Preferably, Formula I is R8 is Preferably, Formula I is R9 is Preferably, Formula I is R10 is Preferably, Formula I is R 11 for Preferably, Formula I is R 12 for 6. A compound or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, cocrystal or deuterated form thereof, wherein the compound is 7. Intermediate compound, which is the following compound 8. A method for preparing a compound according to any one of claims 1 to 5, comprising (1) a) reacting the compound of formula II with sodium acetate at 80°C-120°C for 1-2 hours, adding the compound of formula I, and reacting at 20°C-50°C for 1-8 hours; b) hydrolyzing the compound of formula III obtained in step a) at 50° C.-90° C. for 1-8 hours; c) reacting the product compound of formula IV obtained in step b) with RB 环 -NH2 is reacted at 20°C-50°C for 5-7 hours to obtain a compound of formula V; (2) d) stirring the compound of formula VI, the compound of formula VII and K2CO3 at 20°C-50°C for 20-30 hours, then stirring at -15°C until the color turns orange-yellow, then adding DBU and bromochloromethane at -20°C-0°C, and reacting at 20°C-50°C for 6-24 hours; e) stirring the product compound of formula VIII obtained in step d) at 20° C.-50° C. for 8-12 hours for hydrolysis reaction; f) reacting the product compound of formula IX obtained in step e) with R B环 -NH2 reacts at 20℃-50℃ for 4-8 hours; (3) g) reacting the compound of formula XI with the compound of formula XII at 60° C.-100° C. for 10-14 hours; h) hydrolyzing the compound of formula XIII obtained in step g) at 20° C.-50° C. for 2-6 hours; i) reacting the compound of formula XIV obtained in step h) with R B环 -NH2 reacts at 20℃-50℃ for 10-14 hours; or (4) j) stirring the compound of formula XVI and the compound of formula XVII at 60° C.-100° C. for 10-14 hours; k) stirring the product compound of formula XVIII obtained in step j) at 60° C.-100° C. for hydrolysis reaction for 2-6 hours; l) reacting the product compound of formula XIX obtained in step k) with RB 环 -NH2 was stirred at 20°C-50°C for 10-14 hours; in, R as claimed in any one of claims 1 to 5, RB 环 is the B ring as described in any one of claims 1 to 5, and X is O or S.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 and a pharmaceutically acceptable adjuvant or excipient.

10. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing and / or treating inflammatory diseases, respiratory diseases, skin diseases, or immune system diseases; more preferably, the inflammatory disease is an inflammatory skin disease or a lung inflammatory disease; more preferably, the lung inflammatory disease is asthma or chronic obstructive pulmonary disease; more preferably, the respiratory disease is chronic obstructive pulmonary disease or asthma; more preferably, the skin disease is psoriasis or atopic dermatitis; more preferably, the immune system disease is systemic lupus erythematosus or rheumatoid arthritis.

11. Use of the compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing and / or treating a disease mediated by PDE3 and / or PDE4, an inhibitor of PDE3 or PDE4, or an inhibitor of PDE3 and PDE4.

Images