Novel PDE4 inhibitor and use thereof

Abstract

A novel PDE4 inhibitor and the use thereof; the compound can be used for the treatment of PDE4-mediated diseases.

Description

Novel pde4 inhibitors and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and specifically relates to difluoromethoxyphenyl derivatives with anti-inflammatory and anti-psoriasis effects. BACKGROUND

[0002] Phosphodiesterases (PDEs) belong to the family of hydrolases. Their function is to hydrolyze two intracellular second messenger biologically active cyclic nucleotides, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), into biologically inactive linear nucleotides. Phosphodiesterases (PDEs) are divided into 11 subfamilies of PDEs, which play a key role in regulating cell functions by metabolizing the 3'-cyclic phosphate bond of cAMP and cGMP, and PDE4 is a subtype of PDE.

[0003] PDE4 is a cAMP-specific enzyme that converts the second messenger cAMP into 5'-AMP. On the other hand, cAMP has a profound effect on the functions of inflammatory cell pathways. Elevated intracellular cAMP levels inhibit the activation of T cells, can regulate macrophages and neutrophils, and cause bronchodilation. Increasing intracellular cAMP levels can also inhibit the release of fibrosis, inflammatory cytokines and chemotactic factors, the biological activity of proteases, the production of reactive oxygen species, and the generation of arachidonic acid metabolites. Inhibition of PDE4 increases cAMP levels, thereby relaxing airway smooth muscle and maintaining immune balance.

[0004] The PDE4 target has been developed for the treatment of various inflammatory diseases, including respiratory diseases (chronic obstructive pulmonary disease, asthma), various skin diseases (psoriasis, atopic dermatitis, etc.), and immune system diseases (systemic lupus erythematosus, rheumatoid arthritis, etc.). Common PDE4 inhibitors include Apremilast for moderate to severe psoriasis and rheumatoid arthritis, Crisaborole for mild to moderate psoriasis for systemic / local administration, and Roflumilast for the adjuvant treatment of chronic obstructive pulmonary disease (COPD). Currently, PDE4 inhibitors have become the first-line drug for the treatment of psoriasis and some other chronic inflammatory diseases, and their safe and effective efficacy has been widely recognized.

[0005] Inflammation is a common and frequently-occurring disease that threatens human health. There are many causes of inflammation, and bacteria, viruses, rickettsia, mycoplasma, fungi, etc. can all cause inflammation. Inflammation caused by biological pathogens is also called infection. The human body is complex in structure, and different parts can have different degrees of inflammation. For example, gastrointestinal inflammation, hepatitis, appendicitis, pancreatitis, pharyngitis, prostatitis, vaginitis, periarthritis, otitis media, etc. are all quite representative.

[0006] Psoriasis is an immune-mediated disease. Psoriasis is classified into four types according to clinical features: plaque, pustular, erythrodermic and arthropathic. Among them, plaque is the most common. The pathogenesis of psoriasis is characterized by redness of the skin, large areas of scaling, and water droplet-shaped distribution on various parts of the body. At the same time, it is accompanied by severe itching and burning sensation. Important cells in the pathogenesis of psoriasis are DC, Th17, Th1 and keratinocytes. DC is activated by various stimuli to produce and secrete TNF-a, IL-23 and IL-12. IL-23 induces T cell differentiation into Th17. Activated Th17 cells overproduce IL-17 and IL-22. TNF-a and IL-17 activate keratinocytes, promote epidermal proliferation, promote inflammatory cells (such as neutrophils), and induce the production of antimicrobial peptides (AMP). IL-12 produced by DC also induces Th1 to produce cytokine IFN-g, which exacerbates psoriasis.

[0007] Traditional therapeutic drugs have problems such as poor efficacy and toxic side effects, and the demand for innovative drugs in the clinic is expanding, and more new small molecule compounds are urgently needed.

[0008] SUMMARY

[0009] One or more embodiments of the present application provide a compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal or deuteride thereof

[0010] Formula I

[0011] wherein

[0012] R1and R2are each independently H, and R1and R2are not simultaneously H;

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

[0014] Z is NH or O;

[0015] R3is C6-14 aryl, 5-14 membered heteroaryl, C6-10 cycloalkyl, or 5-14 membered heterocyclyl; optionally, the C6-14 aryl, 5-14 membered heteroaryl, C6-10 cycloalkyl, or 5-14 membered heterocyclyl is substituted with one or more substituents selected from halogen, halogenated C1-C6 alkyl, carbonyl, C1-C6 alkyl, and C1-C6 alkoxy; the 5-14 membered heteroaryl or 5-14 membered heterocyclyl comprises 1-3 heteroatoms selected from N, O and S; preferably, the halogen is F, Cl, Br or I;

[0016] n is 0, 1, 2 or 3;

[0017] Each dashed line independently represents a bond that is present or absent.

[0018] In one or more embodiments, R1is R2is H.

[0019] In one or more embodiments, R1is R2is H.

[0020] In one or more embodiments, R1is R2is

[0021] In one or more embodiments, R1is R2is

[0022] In one or more embodiments, X is N and Y is NH.

[0023] In one or more embodiments, X is N and Y is O.

[0024] In one or more embodiments, X is N and Y is S.

[0025] In one or more embodiments, R3is C6-10 aryl, 5-10 membered heteroaryl, C6-10 cycloalkyl, or 5-10 membered heterocyclyl; optionally, the C6-10 aryl, 5-10 membered heteroaryl, C6-10 cycloalkyl, or 5-10 membered heterocyclyl is substituted with one or more substituents selected from F, carbonyl, C1-C4 alkyl, and C1-C4 alkoxy; the 5-10 membered heteroaryl or 5-10 membered heterocyclyl comprises 1, 2, or 3 heteroatoms selected from N, O, and S.

[0026] In one or more embodiments, wherein R3is

[0027] In one or more embodiments, wherein Formula I is R4is n is 0 or 1.

[0028] In one or more embodiments, wherein Formula I is wherein each R5is independently n is 0 or 1.

[0029] In one or more embodiments, wherein Formula I is wherein R5is

[0030] In one or more embodiments, wherein Formula I is wherein R5is

[0031] In one or more embodiments, wherein Formula I is R6is n is 0 or 1.

[0032] In one or more embodiments, wherein Formula I is R7is n is 0 or 1.

[0033] In one or more embodiments, wherein Formula I is R8is n is 0 or 1.

[0034] In one or more embodiments, wherein Formula I is R9is n is 0 or 1.

[0035] One or more embodiments of the present application provide a compound or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuteride thereof:

[0036] One or more embodiments of the present application provide an intermediate compound for preparing the compounds of the present application, which has the structure as follows:

[0037] One or more embodiments of the present application provide a method for preparing a compound of Formula II,

[0038] wherein

[0039] R1, R2, X, Y, R3, n are as described above;

[0040] The preparation method comprises:

[0041] (1)

[0042] wherein

[0043] a) reacting 3,3-dibromo-1,1,1-trifluoro-2-ketone with sodium acetate at 80-120 °C for 1-2 hours, adding a benzaldehyde solution substituted with R1, R2, difluoromethoxy, reacting at 20-50 °C for 1-8 hours;

[0044] b) hydrolysis of the product obtained in step a) at 50-90 °C for 1-8 hours;

[0045] c) reaction of the product obtained in step b) with R3-(CH2) n -NH2 at 20-50 °C for 5-7 hours;

[0046] (2)

[0047] wherein

[0048] d) reaction of the benzoic acid substituted with R1, R2, difluoromethoxy with L- serine methyl ester hydrochloride and SOCl2 at 0 °C for 10-14 hours;

[0049] e) stirring of the product obtained in step d) with DAST at -78 °C for 3-5 hours, addition of inorganic base and reaction at room temperature for 24 hours;

[0050] f) reaction of the product obtained in step e) with CBrCl3 and DBU at 0 °C for 15-24 hours;

[0051] g) hydrolysis of the product obtained in step f) at 20-50 °C for 1-8 hours;

[0052] h) reaction of the product obtained in step g) with R3-(CH2) n -NH2 at 20-50 °C for 3-8 hours;

[0053] or

[0054] (3)

[0055] reaction of the benzaldehyde substituted with R1, R2, difluoromethoxy with D- cysteine methyl ester hydrochloride, K2CO3 at 20-50 °C for 20-30 hours, then stirring at -15 °C, then addition of DBU and CBrCl3 at -20-0 °C and reaction at 20-50 °C for 12-24 hours;

[0056] i) hydrolysis of the product obtained in step i) at 20-50 °C for 8-12 hours;

[0057] k) reaction of the product obtained in step j) with R3-(CH2) n -NH2 at 20-50 °C for 4-8 hours.

[0058] One or more embodiments of the present application provide a pharmaceutical composition comprising a compound of the present application and a pharmaceutically acceptable adjuvant or excipient.

[0059] One or more embodiments of the present application provide use of a compound of the present application or a pharmaceutical composition of the present application in the manufacture of a medicament for preventing and / or treating an inflammatory disease, a respiratory disease, a skin disease, or an immune system disease.

[0060] One or more embodiments of the present application provide a compound of the present application for use as a medicament.

[0061] One or more embodiments of the present application provide a pharmaceutical composition of the present application for use as a medicament.

[0062] One or more embodiments of the present application provide a compound or composition of the present application for use in preventing and / or treating an inflammatory disease, a respiratory disease, a skin disease, or an immune system disease.

[0063] One or more embodiments of the present application provide a compound or composition of the present application for use in preventing and / or treating a PDE4-mediated disease.

[0064] One or more embodiments of the present application provide a compound or composition of the present application for use in inhibiting PDE4.

[0065] One or more embodiments of the present application provide a method of preventing and / or treating an inflammatory disease, a respiratory disease, a skin disease, or an immune system disease, the method comprising administering to a subject in need thereof a compound or composition of the present application.

[0066] One or more embodiments of the present application provide a method of preventing and / or treating a PDE4-mediated disease, the method comprising administering to a subject in need thereof a compound or composition of the present application.

[0067] One or more embodiments of the present application provide a method of inhibiting PDE4, the method comprising administering to a subject in need thereof a compound or composition of the present application.

[0068] In one or more embodiments, the inflammatory disease is an inflammatory skin disease.

[0069] In one or more embodiments, the respiratory disease is chronic obstructive pulmonary disease or asthma.

[0070] In one or more embodiments, the skin disease is psoriasis or atopic dermatitis.

[0071] In one or more embodiments, the immune system disease is systemic lupus erythematosus or rheumatoid arthritis.

[0072] One or more embodiments of the present application provide use of a compound of the present application or a pharmaceutical composition of the present application in the manufacture of a medicament for preventing and / or treating a PDE4-mediated disease or a PDE4 inhibitor.

[0073] In one or more embodiments, the compounds of the present application have anti-inflammatory and therapeutic effects on psoriasis. Biological experiments show that the compounds of the present application can significantly reduce the number of inflammatory cells in an inflammation model, or reduce the levels of IL-1β, IL-6, TNF-α, IL-17A in cells.

[0074] In one or more embodiments, the results of psoriasis model mouse experiments show that the compounds of the present application can significantly improve the severity of mouse psoriasis thickness, scales, and erythema, indicating that the compounds of the present application can reduce the release of pro-inflammatory factors and the production of pro-inflammatory mediators to inhibit the occurrence and development of inflammatory reactions, and can also protect the skin of mice and have anti-psoriasis effects.

[0075] The following explains the terms used in the technical solutions of the present application. As used in the specification and the appended claims, unless otherwise indicated, the terms of the present application have the following meanings:

[0076] The term "halogen" means fluorine, chlorine, bromine, or iodine.

[0077] The term "amino" means -NH2.

[0078] The term "hydroxy" means -OH.

[0079] "Alkyl" means a straight or branched chain saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, preferably 1 to 8 (e.g., 1, 2, 3, 4, 5, 6, 7, 8) carbon atoms, more preferably 1 to 6 carbon atoms, and even more preferably 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, neopentyl, t-butyl, n-pentyl, i-pentyl, neopentyl, n-hexyl, and various branched isomers thereof; when substituted, the alkyl group can be optionally further substituted with one or more substituents.

[0080] "Alkoxy" means a group formed by the substitution of at least one carbon atom of an alkyl group with an oxygen atom. Non-limiting examples include methoxy, ethoxy, n-propyloxy, i-propyloxy, n-butyloxy, s-butyloxy, t-butyloxy, n-pentyloxy, n-hexyloxy, cyclopropyloxy, and cyclobutyloxy. The definition of alkyl is the same as that described above for "alkyl".

[0081] "Aryl" means a substituted or unsubstituted aromatic ring, which for example can be 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 of 10 to 14 carbon atoms (e.g., 10, 11, 12, 13, 14 carbon atoms), which can be fused or spiro, non-limiting examples include phenyl, naphthyl. The aryl group can be optionally further substituted with one or more substituents.

[0082] "Heteroaryl" means a substituted or unsubstituted aromatic ring, which for example can be a monocyclic ring of 5 to 8 members (e.g., 5, 6, 7, 8 members), a bicyclic ring of 5 to 12 members (e.g., 5, 6, 7, 8, 9, 10, 11, 12 members), or a tricyclic ring of 10 to 14 members (e.g., 10, 11, 12, 13, 14 members), and contains 1 to 6 (e.g., 1, 2, 3, 4, 5, 6) heteroatoms selected from N, O, or S, preferably 5 to 8 membered heteroaryl, 1 to 4 (e.g., 1, 2, 3, 4) of the N, S in the ring of the heteroaryl group optionally oxidized to various oxidation states. The heteroaryl group can be attached at a heteroatom or carbon atom, the heteroaryl group can be fused or spiro, non-limiting examples include, cinnolinyl, furanyl, thienyl, pyranyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, imidazolyl, piperidinyl benzimidazolyl, benzopyridinyl, pyrrolopyridinyl. The heteroaryl group can be optionally further substituted with one or more substituents.

[0083] "Heterocyclyl" or "heterocycle" means a saturated or unsaturated non-aromatic heterocycle, which for example can be a 5- to 10-membered (e.g., 5-, 6-, 7-, 8-, 9-, 10-membered) monocyclic, 5- to 12-membered (e.g., 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-membered) bicyclic, or 10- to 14-membered (e.g., 10-, 11-, 12-, 13-, 14-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. Optionally 1 to 4 (e.g., 1, 2, 3, 4) of the N, S in the ring of the "heterocyclyl" or "heterocycle" can be oxidized into various oxidation states; the "heterocyclyl" or "heterocycle" can be attached at a heteroatom or carbon atom; the "heterocyclyl" or "heterocycle" can be a bridged or spirocyclic ring. Non-limiting examples of "heterocyclyl" or "heterocycle" include oxiranyl, oxetanyl, aziridinyl, oxetanyl, thietanyl, 1,3-dioxolanyl, 1,4-dioxolanyl, 1,3-dioxananyl, azepanyl, oxepanyl, thiepanyl, dioxepanyl, pyridinyl, piperidinyl, homopiperidinyl, furanyl, thienyl, pyranyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, homopiperazinyl, imidazolyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, thioxanyl, 1,3-dithianyl, dihydrofuranyl, dihydropyranyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, tetrahydrothiopyranyl, tetrahydropyrrolyl, tetrahydroimidazolyl, tetrahydrothiazolyl, tetrahydropyranyl, benzimidazolyl, benzopyridinyl, pyrrolopyridinyl, benzodihydrofuranyl, 2-pyrrolinyl, 3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydrothienyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 1,2,3,4-tetrahydroisoquinolinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, azabicyclo[2.2.2]hexanyl, 3H-indolizinyl, quinolizinyl, N-pyridinylurea, 1,1-dioxothiomorpholinyl, azabicyclo[3.2.1]octanyl, azabicyclo[5.2.0]nonyl, oxatricyclo[5.3.1.1]dodecanyl, azadamantanyl, and oxaspiro[3.3]heptanyl. The "heterocyclyl" or "heterocycle" can be optionally further substituted with one or more substituents.

[0084] "Cycloalkyl" means a saturated cyclic hydrocarbon group, which for example can be a 6- to 10-carbon atom (e.g., 6-, 7-, 8-, 9-, 10-carbon atom) monocyclic, bicyclic, or polycyclic ring, preferably 6- to 8-carbon atom ring. Non-limiting examples of "cycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. When the cycloalkyl is substituted, it can be optionally further substituted with one or more substituents.

[0085] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salt thereof means a salt of a compound of the present application which retains the biological effectiveness and properties of the free acids or free bases and is obtained by reaction of the free acid with a non-toxic inorganic or organic base, or the free base with a non-toxic inorganic or organic acid.

[0086] "Pharmaceutical composition" means a mixture of one or more compounds of the present application, pharmaceutically acceptable salts or prodrugs thereof, and other chemical components, such as pharmaceutically acceptable carriers, excipients, and / or one or more other therapeutic agents.

[0087] "Carrier" means a material that does not itself induce the production of antibodies to it, and which does not have an adverse effect on the activity and properties of a given compound.

[0088] "Excipient" means 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 and its derivatives, gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, binders, and disintegrating agents.

[0089] "Prodrug" means a compound that is converted into a biologically, pharmaceutically or therapeutically active compound of the present application after administration to a subject. Prodrugs of the present application are prepared by modifying the amino or carboxyl groups of the compounds of the present application in such a way that the modifications are cleaved in vivo to give the parent compound. When a prodrug of the present application is administered to a mammalian subject, the prodrug is cleaved to form the free amino or carboxyl groups.

[0090] "Cocrystal" means a crystal formed by the combination of an active pharmaceutical ingredient (API) and a cocrystal former (CCF) under the action of hydrogen bonds or other non-covalent bonds, wherein the pure state of the API and the CCF are both solid at room temperature, and there is a fixed stoichiometric ratio between the components. Cocrystals are a kind of multi-component crystals, including binary cocrystals formed between two neutral solids, and multi-component cocrystals formed between a neutral solid and a salt or a solvate.

[0091] "Stereoisomer" means isomers that have the same molecular formula but differ in the arrangement of atoms in space. Stereoisomers include enantiomers (mirror image isomers), diastereomers (geometric isomers), and conformers.

[0092] "Optional" or "optionally" or "selective" or "selectively" means that the subsequently described event or circumstance can or can not occur, and this description includes instances in which the event or circumstance occurs and instances in which it does not. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group can or can not be present, and this description includes instances in which the heterocyclyl is substituted with alkyl, and instances in which the heterocyclyl is not substituted with alkyl. BRIEF DESCRIPTION OF DRAWINGS

[0093] Figures 1 and 2 represent the phenotypic presentation of the dorsal skin of each group (n=8, male) treated with Compound A5 and Compound D2 respectively in the active example 8 for 7 days.

[0094] Figures 3 and 4 represent the daily monitoring of the weight chart of Compound A5 and Compound D2 in the active example 8, where the erythema was monitored daily, scored and thickness scored according to PASI, the score ranged from 0 to 4, the total score was monitored daily according to PASI, the cumulative score ranged from 0 to 12 points.

[0095] Figures 5 and 6 represent the pathological changes observed under the microscope of the skin tissue of Compound A5 and Compound D2 in the active example 8.

[0096] Figures 7 and 8 represent the effect of Compound A5 and Compound D2 on the spleen in the mouse psoriasis model in the active example 8.

[0097] Figures 9 and 10 represent the Ki-67 antibody immunohistochemical staining of the skin tissue of Compound A5 and Compound D2 in the mouse psoriasis model in the active example 8.

[0098] Figures 11 and 12 represent the inhibition of the expression of inflammatory factors in the skin tissue of Compound A5 and Compound D2 in the mouse psoriasis model in the active example 8. DETAILED DESCRIPTION

[0099] The present application will be further described in conjunction with specific examples. These examples are for illustrative purposes only and are not intended to limit the scope and spirit of the present application. All other examples obtained by those of ordinary skill in the art based on the examples in the present application without creative work are within the scope of protection of the present application.

[0100] Example 1 Synthesis of a series of compounds

[0101] (1) Synthesis of compound 2:

[0102] Weigh 3 g of compound 3,4-dihydroxybenzaldehyde (21.72 mmol, 1.0 equiv.) and 6.91 g of sodium carbonate (65.16 mmol, 3.0 equiv.) and dissolve in 30 mL of N,N-dimethylformamide, finally add 3.44 g of ethyl difluorochloroacetate (21.72 mmol, 1.0 equiv.), react at 80°C for 8 hours, after the raw material is completely reacted by TLC, quench the reaction with water. Extract with ethyl acetate 3 times, wash the organic phase with saturated sodium chloride solution. Concentrate the organic phase and purify by flash column chromatography to obtain 1.4 g of white solid product (compound 2) with a yield of 34%.

[0103] The obtained compound 2 was identified by nuclear magnetic resonance spectrum, mass spectrometry, and the identification result was as follows: 1 H NMR (500MHz, CDCI3) δ 9.90 (s, 1H), 7.53 (d, J = 2.0 Hz, 1H), 7.44 (dd, J = 8.5, 2.0 Hz, 1H), 7.26 (d, J = 8.5 Hz, 1H), 6.66 (t, J = 73.5 Hz, 1H). ESI-HRMS m / z: calculated C8H6O3F2 + [M+Na] + , 211.0177; found 211.0166.

[0104] (2) Synthesis of compound 3:

[0105] Compound 2.1 g (11.16 mmol, 1.0 equiv.) was weighed, 3.09 g of potassium carbonate (22.32 mmol, 2.0 equiv.) was added, 20 mL of N, N-dimethylformamide was added, and finally 2.26 g of bromomethylcyclopropane (16.74 mmol, 1.5 equiv.) was added, and the reaction was carried out at 80°C for 8 hours. TLC was used to detect that the raw material was completely reacted, and water was added to quench the reaction. Ethyl acetate was used for extraction for 3 times, and the organic phase was washed with saturated sodium chloride solution. The organic phase was concentrated and column chromatography was used to obtain 2.3 g of yellow oily product (compound 3), and the yield was 85%.

[0106] The obtained compound 3 was identified by nuclear magnetic resonance spectrum, mass spectrometry, and the identification result was as follows: 1 H NMR (500MHz, DMSO-d6) δ 9.94 (s, 1H), 7.59-7.54 (m, 2H), 7.39 (d, J = 8.0 Hz, 1H), 7.28 (t, J = 73.5 Hz, 1H), 3.98 (d, J = 7.0 Hz, 2H), 1.30-1.21 (m, 1H), 0.61-0.55 (m, 2H), 0.39-0.33 (m, 2H). ESI-MS m / z: calculated C 12 H 13 O3F2 + [M+H] + , 243.1; found 243.1.

[0107] (3) Synthesis of compound 4:

[0108] Weigh 4.46 g of 3,3-dibromo-1,1,1-trifluoro-2-ketone (16.52 mmol, 2.0 equiv.) and 2.7 g of sodium acetate (33.04 mmol, 4.0 equiv.) into a reaction bottle, dissolve in a proper amount of water, stir at 100°C for 1 hour, cool to room temperature, then add 2 g of compound 3 (8.26 mmol, 1.0 equiv.) in methanol and 4 mL of ammonia water into the above solution, stir at room temperature, and when the raw material is no longer reduced by TLC detection, remove the methanol by rotary evaporation. Extract with ethyl acetate for 3 times, wash the organic phase with saturated sodium chloride solution once, and dry the organic phase over anhydrous sodium sulfate. Concentrate the organic phase and purify by flash column chromatography to obtain 1.37 g of yellow solid compound 4 with a yield of 48%.

[0109] The obtained compound 4 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1 H NMR (400MHz, CDC13) δ 12.56 (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 for C 15 H 14 O2N2F5 + [M+H] + , 349.1; found 349.1.

[0110] (4) Synthesis of compound 5:

[0111] Weigh 415 mg of compound 4 (1.19 mmol, 1.0 equiv.) and dissolve in a 1:1 mixture of ethanol and water, add 953 mg of sodium hydroxide (23.80 mmol, 20 equiv.), stir the reaction at 80°C, monitor by TLC, and after the reaction is complete, remove the ethanol by rotary evaporation, add a proper amount of water, adjust the pH to 6 with dilute hydrochloric acid. Filter to obtain a solid, and dry to obtain 322 mg of yellow solid compound 5 with a yield of 83%.

[0112] The obtained compound 5 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1H NMR (400MHz, DMSO-d6) δ 13.28 (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), OH (1H, not observed). ESI-HRMS m / z: calcd for C 15 H 14 O4N2F2Na + [M+Na] + , 347.0814; found, 347.0800.

[0113] (5) Synthesis of compounds A1-A17:

[0114] Weigh 50 mg of compound 5 (0.15 mmol, 1.0 equiv.) dissolved in N, N- dimethylformamide, add 59 mg of HATU (0.15 mmol, 1.0 equiv.) and 60 mg of DIPEA (0.45 mmol, 3.0 equiv.), stir at room temperature for 20 min, add the corresponding amine reagent (0.15 mmol, 1.0 equiv.), react at room temperature for 6 h. Monitor with TLC, and when the raw material is completely reacted, quench with water, extract with ethyl acetate, wash once with saturated sodium chloride solution, dry the ethyl acetate layer with anhydrous sodium sulfate, concentrate the solution, and purify by column chromatography to obtain compounds A1-A17.

[0115] The compounds A1-A17 are arranged in sequence as follows:

[0116] Synthesis of compound A1 in Example 2:

[0117] The synthesis steps are referred to Example 1, and the amine reagent of the last step reaction is selected as 2-aminothiazole to obtain compound A1, yield: 50%; 1 H NMR (500MHz, DMSO-d6) δ 13.27 (br, 1H), 11.54 (s, 1H), 8.19 (s, 1H), 7.82 (s, 1H), 7.65 (s, 1H), 7.53 (d, J = 3.5 Hz, 1H), 7.30 (s, 1H), 7.25 (d, J = 3.5 Hz, 1H), 7.15 (t, J = 74.5 Hz, 1H), 3.99 (d, J = 7.0 Hz, 2H), 1.38-1.24 (m, 1H), 0.66-0.56 (m, 2H), 0.43-0.34 (m, 2H).13 C NMR (151 MHz, DMSO-d6) δ 160.1, 157.9, 150.1, 146.0, 140.3, 137.7, 134.5, 127.9, 123.3, 121.3, 117.9, 116.7 (t, J = 256.5 Hz), 113.6, 111.4, 73.1, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 18 H 17 O3N4F2S + [M+H] + , 407.0984; found 407.0964.

[0118] Synthesis of compound A2 in Example 3:

[0119] The synthetic procedure was referred to Example 1, and the amine reagent in the last step reaction was chosen as 4-aminopyridine to give compound A2 in yield of 58%; 1 H NMR (400 MHz, DMSO-d6) δ 13.26 (br, 1H), 10.17 (br, 1H), 8.46 (d, J = 5.6 Hz, 2H), 8.08 (s, 1H), 7.88 (d, J = 4.8 Hz, 2H), 7.81 (s, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.31 (d, J = 8.4 Hz, 1H), 7.16 (t, J = 74.4 Hz, 1H), 3.99 (d, J = 6.8 Hz, 2H), 1.36 - 1.26 (m, 1H), 0.66 - 0.56 (m, 2H), 0.42 - 0.34 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 161.6, 150.2 (2 x C), 150.1, 145.7, 145.4, 140.2, 136.2, 128.0, 123.0, 121.3, 118.1, 116.7 (t, J = 256.4 Hz), 113.9 (2 x C), 111.4, 73.2, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 20 H 19 O3N4F2 + [M+H] + , 401.1420; found 401.1402.

[0120] Synthesis of compound A3 in Example 4:

[0121] The synthetic procedure was referred to Example 1, and the amine reagent in the last step reaction was chosen as 2,6-difluorobenzylamine to give compound A3 in yield of 7%.1 H NMR (600 MHz, MeOD-d4) δ 7.71 (s, 1H), 7.65 (d, J = 1.8 Hz, 1H), 7.37 (d, J = 7.2 Hz, 1H), 7.27-7.20 (m, 1H), 7.17 (d, J = 8.4 Hz, 1H), 6.82 (s, 2H), 6.77 (t, J = 75.0 Hz, 1H), 4.58 (s, 2H), 3.64 (s, 2H), 1.21-1.12 (m, 1H), 0.56 (d, J = 7.2 Hz, 2H), 0.23 (s, 2H), NH (2H, not observed). 13 C NMR (151 MHz, MeOD-d4) δ 164.7, 162.9 (dd, J = 247.5, 7.5 Hz, 2 x C), 152.2, 147.7, 142.4, 137.5, 131.1 (t, J = 10.5 Hz), 129.3, 123.3, 122.4, 118.7, 117.9 (t, J = 256.5 Hz), 114.5 (t, J = 18.0 Hz), 112.6, 112.2 (dd, J = 21.0, 6.0 Hz, 2 x C), 74.6, 31.5, 10.8, 3.5 (2 x C). ESI-HRMS m / z: calcd for C 22 H 20 O3N3F4 + [M+H] + , 450.1435; found 450.1408.

[0122] Synthesis of compound A4 of example 5:

[0123] Synthesis procedure as in example 1, amine reagent for last step reaction was chosen as 2-methylaminopyrimidine (CAS number: 75985-45-4) to give compound A4 in 14% yield; 1 H NMR (400 MHz, DMSO-d6) δ 13.02 (br, 1H), 8.77 (d, J = 4.8 Hz, 2H), 8.46 (br, 1H), 7.82 (s, 1H), 7.76 (s, 1H), 7.63 (d, J = 8.0 Hz, 1H), 7.40 (t, J = 4.8 Hz, 1H), 7.28 (d, J = 8.4 Hz, 1H), 7.14 (t, J = 74.4 Hz, 1H), 4.68 (d, J = 6.0 Hz, 2H), 3.97 (d, J = 6.8 Hz, 2H), 1.33-1.25 (m, 1H), 0.62-0.58 (m, 2H), 0.40-0.36 (m, 2H). 13C NMR (101 MHz, DMSO-d6) δ 166.9, 162.2, 157.4 (2 x C), 150.1, 147.1, 144.9, 140.0, 136.9, 121.4, 120.9, 119.9, 117.7, 116.7 (t, J = 256.4 Hz), 111.1, 73.2, 44.5, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 20 H 20 O3N5F2 + [M+H] + , 416.1529; found 416.1530.

[0124] Synthesis of compound A5 of Example 6:

[0125] Synthesis procedure was referred to Example 1, and the amine reagent of the last step reaction was chosen as 2,5-difluorobenzylamine to give compound A5, yield: 56%; 1 H NMR (600 MHz, MeOD-d4) δ 7.74 (s, 1H), 7.72 (d, J = 2.4 Hz, 1H), 7.42 (dd, J = 7.8, 1.8 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H), 6.99 - 6.91 (m, 3H), 6.77 (t, J = 75.6 Hz, 1H), 4.47 (s, 2H), 3.69 (d, J = 5.4 Hz, 2H), 1.20 - 1.15 (m, 1H), 0.57 - 0.53 (m, 2H), 0.22 (d, J = 4.8 Hz, 2H), NH (2H, not observed). 13 C NMR (151 MHz, MeOD-d4) δ 165.1, 160.0 (dd, J = 240.0, 3.0 Hz), 157.9 (dd, J = 240.0, 3.0 Hz), 152.2, 147.9, 142.5, 137.3, 129.3, 128.5 (dd, J = 18.0, 7.5 Hz), 123.4, 122.5, 118.7, 117.9 (t, J = 256.5 Hz), 117.4 (dd, J = 24.0, 4.5 Hz), 116.9 (dd, J = 33.0, 9.0 Hz), 116.3 (dd, J = 33.0, 9.0 Hz), 112.6, 74.7, 37.2, 10.8, 3.5 (2 x C). ESI-HRMS m / z: calcd for C 22 H 20 O3N3F4 + [M+H] + , 450.1435; found 450.1413.

[0126] Synthesis of compound A6:

[0127] The synthetic procedure was referenced to example 1, and the amine reagent of the last step reaction was selected as 2,4-difluorobenzylamine to give compound A6 in yield of 53%; 1 H NMR (500 MHz, MeOD-d4) δ 7.73 (s, 1H), 7.69 (d, J = 1.5 Hz, 1H), 7.47-7.42 (m, 1H), 7.34 (q, J = 7.5, 7.0 Hz, 1H), 7.21 (d, J = 8.0 Hz, 1H), 6.87-6.80 (m, 2H), 6.80 (t, J = 75.5 Hz, 1H), 4.54 (s, 2H), 3.84 (d, J = 6.0 Hz, 2H), 1.27-1.24 (m, 1H), 0.65-0.55 (m, 2H), 0.35-0.28 (m, 2H), NH (2H, not observed). 13 C NMR (151 MHz, MeOD-d4) δ 165.2, 163.7 (dd, J = 246, 12.0 Hz), 162.2 (dd, J = 248, 12.0 Hz), 152.2, 147.8, 142.5, 137.6, 132.0 (dd, J = 9.0, 6.0 Hz), 129.4, 123.4, 122.9 (dd, J = 15.0, 4.5 Hz), 122.3, 118.9, 117.9 (t, J = 256.5 Hz), 112.7, 112.1 (dd, J = 25.5, 4.5 Hz), 104.5 (t, J = 27.0 Hz), 74.9, 37.1, 10.9, 3.5 (2 x C). ESI-HRMS m / z: calcd for C 22 H 20 O3N3F4 + [M+H] + , 450.1435; found 450.1415.

[0128] Synthesis of compound A7:

[0129] The synthetic procedure was referenced to example 1, and the amine reagent of the last step reaction was selected as 3,4-difluorobenzylamine to give compound A7 in yield of 23%; 1H NMR (400 MHz, DMSO-d6) δ 13.02 (br, 1H), 8.66 (br, 1H), 7.81 (s, 1H), 7.75 (s, 1H), 7.61 (d, J = 8.0 Hz, 1H), 7.39 - 7.30 (m, 2H), 7.27 (d, J = 8.4 Hz, 1H), 7.17 - 7.14 (m, 1H), 7.12 (t, J = 74.4 Hz, 1H), 4.42 (d, J = 6.4 Hz, 2H), 3.94 (d, J = 7.2 Hz, 2H), 1.33 - 1.24 (m, 1H), 0.64 - 0.54 (m, 2H), 0.38 - 0.32 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 162.3, 150.1, 149.2 (dd, J = 244, 13 Hz), 148.3 (dd, J = 243, 16 Hz), 145.0, 140.0, 137.8, 136.8, 128.2, 123.9 (dd, J = 6.0, 3.0 Hz), 121.3, 120.9, 117.7, 117.1 (d, J = 17 Hz), 116.6 (t, J = 256.5 Hz), 116.3 (d, J = 17 Hz), 111.1, 73.1, 41.0, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 20 O3N3F4 + [M+H] + , 450.1435; found 450.1405.

[0130] Example 9 Synthesis of compound A8:

[0131] Synthesis procedure was referred to Example 1, the amine reagent of last step reaction was selected as aniline, to obtain compound A8, yield: 23%; 1 H NMR (500 MHz, MeOD-d4) δ 7.83 (s, 1H), 7.76 - 7.65 (m, 3H), 7.52 (d, J = 8.5 Hz, 1H), 7.35 (t, J = 8.0 Hz, 2H), 7.24 (d, J = 8.0 Hz, 1H), 7.13 (t, J = 7.5 Hz, 1H), 6.83 (t, J = 75.0 Hz, 1H), 4.00 (d, J = 7.0 Hz, 2H), 1.38 - 1.29 (m, 1H), 0.68 - 0.64 (m, 2H), 0.42 - 0.39 (m, 2H), NH (2H, not observed). 13C NMR (151 MHz, MeOD-d4) δ 163.2, 152.3, 148.0, 142.6, 139.5, 130.8, 129.9 (2 x C), 129.4, 125.3, 123.4, 122.9, 121.5 (2 x C), 119.3, 118.0 (t, J = 256.5 Hz), 112.9, 75.0, 11.0, 3.6 (2 x C). ESI-HRMS m / z: calcd for C 21 H 19 O3N3F2Na + [M+Na] + , 422.1287; found 422.1283.

[0132] Synthesis of compound A9, Example 10:

[0133] Synthesis procedure as in Example 1, with the amine reagent in the last step reaction selected as 2-(aminomethyl)naphthalene (CAS Number: 2018-90-8) to give compound A9 in 81% yield; 1 H NMR (600 MHz, DMSO-d6) δ 8.71 (s, 1H), 7.90-7.83 (m, 3H), 7.82 (s, 1H), 7.78 (s, 2H), 7.62 (d, J = 7.8 Hz, 1H), 7.53-7.43 (m, 3H), 7.25 (s, 1H), 7.13 (t, J = 74.4 Hz, 1H), 4.63 (d, J = 6.0 Hz, 2H), 3.95 (d, J = 6.6 Hz, 2H), 1.32-1.24 (m, 1H), 0.62-0.55 (m, 2H), 0.40-0.29 (m, 2H), NH (not observed). 13 C NMR (151 MHz, DMSO-d6) δ 161.8, 150.1, 145.3, 140.0, 137.6, 132.9, 132.1, 129.7, 128.4, 127.8, 127.5 (2 x C), 127.3, 126.2, 126.1, 125.6, 125.4, 121.3, 117.8, 116.7 (t, J = 258.0 Hz), 111.2, 73.1, 42.0, 10.0, 3.0 (2 x C). ESI-HRMS m / z: calcd for C 26 H 23 O3N3F2Na + [M+Na] + , 486.1600; found 486.1582.

[0134] Synthesis of compound A10, Example 11:

[0135] The synthesis procedure was referred to Example 1, and the amine reagent for the last step reaction was selected as 2-methoxy-5-(aminomethyl)pyridine (CAS No.: 262295-96-5) to give compound A10 in a yield of 64%; 1 H NMR (600 MHz, DMSO-d6) δ 13.06 (br, 1H), 8.57 (s, 1H), 8.10 (s, 1H), 7.79 (s, 1H), 7.73 (s, 1H), 7.65 (dd, J = 8.4, 2.4 Hz, 1H), 7.59 (d, J = 8.4 Hz, 1H), 7.24 (d, J = 8.4 Hz, 1H), 7.12 (t, J = 74.4 Hz, 1H), 6.75 (d, J = 8.4 Hz, 1H), 4.37 (d, J = 6.0 Hz, 2H), 3.92 (d, J = 6.6 Hz, 2H), 3.81 (s, 3H), 1.32 - 1.24 (m, 1H), 0.62 - 0.56 (m, 2H), 0.38 - 0.32 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 162.7, 162.1, 150.1, 145.9, 145.0, 140.1, 139.0, 136.9, 128.5, 128.3, 121.4, 121.0, 117.8, 116.7 (t, J = 256.5 Hz), 111.2, 110.2, 73.2, 53.1, 39.0, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 22 O4N4F2Na + [M+Na] + , 467.1501; found 467.1485.

[0136] Synthesis of compound A11

[0137] The synthesis procedure was referred to Example 1, and the amine reagent for the last step reaction was selected as 2-methoxy-5-(aminomethyl)pyridine (CAS No.: 262295-96-5) to give compound A10 in a yield of 64%; 1H NMR (500 MHz, DMSO-d6) δ 13.04 (br, 1H), 8.66 (s, 1H), 8.61 (s, 1H), 8.58 (s, 1H), 8.53 (d, J = 2.5 Hz, 1H), 7.83 (s, 1H), 7.75 (s, 1H), 7.61 (d, J = 7.0 Hz, 1H), 7.28 (s, 1H), 7.13 (t, J = 74.5 Hz, 1H), 4.62 (d, J = 6.0 Hz, 2H), 3.96 (d, J = 7.0 Hz, 2H), 1.35 - 1.23 (m, 1H), 0.63 - 0.52 (m, 2H), 0.42 - 0.32 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 162.5, 154.5, 150.1, 145.0, 143.9, 143.4, 143.1, 140.0, 136.7, 128.3, 121.4, 121.1, 117.7, 116.7 (t, J = 258.0 Hz), 111.2, 73.2, 42.0, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 20 H 19 O3N5F2Na + [M+Na] + , 438.1348; found 438.1304.

[0138] Synthesis of compound A12 of example 13:

[0139] Synthesis procedure as in example 1, the amine reagent for the last step reaction was chosen to be benzylamine to give compound A12 in 57% yield; 1 H NMR (400 MHz, DMSO-d6) δ 13.01 (br, 1H), 8.52 (t, J = 6.4 Hz, 1H), 7.82 (s, 1H), 7.73 (d, J = 1.6 Hz, 1H), 7.59 (dd, J = 8.4, 1.6 Hz, 1H), 7.34 - 7.30 (m, 4H), 7.28 - 7.25 (m, 1H), 7.25 - 7.20 (m, 1H), 7.14 (t, J = 74.4 Hz, 1H), 4.46 (d, J = 6.4 Hz, 2H), 3.96 (d, J = 6.8 Hz, 2H), 1.32 - 1.28 (m, 1H), 0.65 - 0.57 (m, 2H), 0.39 - 0.34 (m, 2H). 13C NMR (101 MHz, DMSO-d6) δ 162.2, 150.1, 144.9, 140.1, 140.0, 137.0, 128.4, 128.3 (2 x C), 127.4 (2 x C), 126.7, 121.3, 120.9, 117.7, 116.7 (t, J = 258.0 Hz), 111.1, 73.1, 41.8, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 21 O3N3F2Na + [M+Na] + 436.1443; found 436.1435.

[0140] Synthesis of compound A13, Example 14:

[0141] The synthesis was performed according to the procedure described in Example 1, with the exception that the amine reagent in the last step was 2-aminomethylpyridine (CAS Number: 3731-51-9) to afford compound A13 in 46% yield. 1 H NMR (400 MHz, DMSO-d6) δ 13.02 (br, 1H), 8.59 (br, 1H), 8.53-8.50 (m, 1H), 7.84 (s, 1H), 7.76-7.70 (m, 2H), 7.60 (d, J = 8.0 Hz, 1H), 7.33-7.25 (m, 3H), 7.14 (t, J = 74.4 Hz, 1H), 4.56 (d, J = 6.0 Hz, 2H), 3.97 (d, J = 6.8 Hz, 2H), 1.34-1.29 (m, 1H), 0.63-0.58 (m, 2H), 0.40-0.35 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 162.3, 158.7, 150.1, 148.8, 144.9, 140.0, 136.9, 136.7, 128.3, 122.0, 121.4, 121.0, 120.9, 117.7, 116.7 (t, J = 258.0 Hz), 111.2, 73.1, 43.8, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 21 H 20 O3N4F2Na + [M+Na] + 437.1396; found 437.1382.

[0142] Synthesis of compound A14, Example 15:

[0143] The synthetic procedure was referred to Example 1, and the amine reagent for the last step reaction was chosen as 4-aminomethyltetrahydropyran (CAS No.: 130290-79-8) to give compound A14 in 55% yield; 1 H NMR (600 MHz, DMSO-d6) δ 12.98 (br, 1H), 7.98 (s, 1H), 7.77 (s, 1H), 7.74 (s, 1H), 7.59 (d, J = 7.8 Hz, 1H), 7.27 (d, J = 7.8 Hz, 1H), 7.06 (t, J = 74.4 Hz, 1H), 3.93 (d, J = 5.4 Hz, 2H), 3.82 (d, J = 10.2 Hz, 2H), 3.23 (t, J = 11.4 Hz, 2H), 3.18-3.08 (m, 2H), 1.75 (s, 1H), 1.53 (d, J = 12.6 Hz, 2H), 1.30-1.25 (m, 1H), 1.21-1.10 (m, 2H), 0.59 (q, J = 5.4 Hz, 2H), 0.35 (s, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 162.2, 150.1, 144.8, 140.0, 137.2, 128.3, 121.3, 120.6, 117.7, 116.7 (t, J = 258.0 Hz), 111.2, 73.1, 66.8 (2 x C), 43.9, 35.1, 30.5 (2 x C), 10.0, 3.0 (2 x C). ESI-HRMS m / z: calcd for C 21 H 25 O4N3F2Na + [M+Na] + , 444.1705; found 444.1693.

[0144] Example 16 Synthesis of compound A15:

[0145] The synthetic procedure was referred to Example 1, and the amine reagent for the last step reaction was chosen as 1-methyl-4-aminopyrazole (CAS No.: 69843-13-6) to give compound A15 in 40% yield; 1H NMR (500 MHz, DMSO-d6) δ 13.08 (br, 1H), 9.99 (s, 1H), 8.01 (s, 1H), 7.90 (s, 1H), 7.79 (s, 1H), 7.66 (d, J = 13.5 Hz, 2H), 7.30 (t, J = 4.0 Hz, 1H), 7.15 (t, J = 74.5 Hz, 1H), 3.99 (d, J = 7.0 Hz, 2H), 3.82 (s, 3H), 1.36 - 1.27 (m, 1H), 0.66 - 0.56 (m, 2H), 0.44 - 0.31 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 159.3, 150.1, 145.0, 140.0, 136.8, 130.3, 129.7, 128.2, 121.6, 121.4, 121.3, 117.9, 116.7 (t, J = 258.0 Hz), 111.3, 73.2, 38.7, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 19 H 19 O3N5F2Na + [M+Na] + , 426.1348; found 426.1337.

[0146] Synthesis of compound A16

[0147] Synthesis procedure as in example 1, the amine reagent for the last step reaction was chosen to be N-methyl-3-aminopyrazole (CAS number: 1904-31-0) to give compound A16 in 62% yield; 1 H NMR (500 MHz, DMSO-d6) δ 13.08 (br, 1H), 9.99 (s, 1H), 8.01 (s, 1H), 7.90 (s, 1H), 7.79 (s, 1H), 7.66 (d, J = 13.5 Hz, 2H), 7.30 (t, J = 4.0 Hz, 1H), 7.15 (t, J = 74.5 Hz, 1H), 3.99 (d, J = 7.0 Hz, 2H), 3.82 (s, 3H), 1.36 - 1.27 (m, 1H), 0.66 - 0.56 (m, 2H), 0.44 - 0.31 (m, 2H). 13C NMR (151 MHz, DMSO-d6) δ 159.5, 150.1, 146.4, 145.2, 140.1, 136.1, 131.2, 128.1, 121.7, 121.3, 117.8, 116.7 (t, J = 256.5 Hz), 111.4, 96.5, 73.2, 38.3, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 19 H 19 O3N5F2Na + [M+Na] + , 426.1348; found 426.1348.

[0148] Synthesis of compound A17 in Example 18:

[0149] Synthetic procedure was referred to Example 1, and the amine reagent of the last step reaction was chosen as 5-aminoindole to give compound A17 in yield of 32%; 1 H NMR (400 MHz, DMSO-d6) δ 13.13 (br, 1H), 11.04 (br, 1H), 9.60 (s, 1H), 8.02 (s, 1H), 7.96 (s, 1H), 7.84 (d, J = 1.6 Hz, 1H), 7.68 (dd, J = 8.4, 1.6 Hz, 1H), 7.45 - 7.41 (m, 1H), 7.39 - 7.27 (m, 3H), 7.16 (t, J = 74.4 Hz, 1H), 6.40 (s, 1H), 3.99 (d, J = 6.8 Hz, 2H), 1.36 - 1.29 (m, 1H), 0.66 - 0.57 (m, 2H), 0.44 - 0.35 (m, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 160.3, 150.2, 145.0, 140.1, 137.3, 132.9, 130.7, 128.3, 127.5, 126.0, 121.4, 121.3, 117.8, 116.8 (t, J = 256.3 Hz), 115.6, 111.4, 111.3, 111.1, 101.1, 73.2, 10.1, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 23 H 20 O3N4F2Na + [M+Na] + , 461.1396; found 461.1394.

[0150] Synthesis of compounds in series B in Example 19

[0151] (1) Synthesis of compound 7:

[0152] Dissolve 1.0 g of compound 3 (4.13 mmol, 1.0 equiv.) in a mixture of methanol and water, add 1.32 g of sodium hydroxide (33.04 mmol, 8.0 equiv.) and 1.32 g of 30% hydrogen peroxide solution (20.65 mmol, 5.0 equiv.), stir at 50 °C, monitor by TLC. Then remove methanol by rotary evaporation, adjust pH to 6 with dilute hydrochloric acid, wash the solid with water for 3 times, to get intermediate 7 (867.5 mg) with a yield of 89%.

[0153] The obtained compound 7 is identified by nuclear magnetic resonance spectroscopy, mass spectrometry, and the identification result is: 1 H NMR (400 MHz, CDC13) δ 7.74 (dd, J = 8.4, 2.0 Hz, 1H), 7.67 (d, J = 2.0 Hz, 1H), 7.25 (d, J = 8.4 Hz, 1H), 6.74 (t, J = 74.8 Hz, 1H), 3.95 (d, J = 6.8 Hz, 2H), 1.37-1.30 (m, 1H), 0.72-0.63 (m, 2H), 0.42-0.35 (m, 2H), OH (1H, not observed). ESI-MS m / z: calculated C 12 H 13 O4F2 + [M+Na] + , 259.0; found 259.0.

[0154] (2) Synthesis of compound 8:

[0155] Dissolve compound 7 (0.57 mmol, 1.0 equiv.) in thionyl chloride and stir at room temperature for 0.5 hours. Remove the solvent using rotary evaporation to get the acyl chloride intermediate. Add triethylamine (1.14 mmol, 2.0 equiv.) to a solution of L-methionine methyl ester hydrochloride (0.57 mmol, 1.0 equiv.) in dichloromethane, stir the mixture for 0.5 hours. Add the acyl chloride intermediate in dichloromethane dropwise at 0 °C. Stir the reaction mixture for 12 hours. Concentrate the reaction, purify by column chromatography to get compound 8. Yield 45%.

[0156] The obtained compound 8 is identified by nuclear magnetic resonance spectroscopy, mass spectrometry, and the identification result is: 1H NMR (400 MHz, CDC13) δ 7.47 (d, J = 2.0 Hz, 1H), 7.32 (dd, J = 8.4, 2.0 Hz, 1H), 7.20 - 7.13 (m, 2H), 6.69 (t, J = 75.2 Hz, 1H), 4.86 - 4.80 (m, 1H), 4.08 (dd, J = 11.6, 3.6 Hz, 1H), 4.02 (dd, J = 11.6, 3.6 Hz, 1H), 3.91 (d, J = 6.8 Hz, 2H), 3.81 (s, 3H), 2.51 (br, 1H), 1.33 - 1.27 (m, 1H), 0.68 - 0.60 (m, 2H), 0.38 - 0.31 (m, 2H). ESI-HRMS m / z: calcd for C 16 H 20 O6NF2 + [M+H] + , 360.1253; found 360.1261.

[0157] (3) Synthesis of compound 9:

[0158] Diethylamine sulfide trifluoride (20.51 mmol, 3.5 equiv.) was added dropwise to a solution of intermediate 8 (5.86 mmol, 1.0 equiv.) in dichloromethane. After stirring at -78 °C for 4 hours, potassium carbonate was added. The reaction mixture was then allowed to warm to room temperature and stirred for 24 hours. After addition of water, the organic layer was extracted with dichloromethane, washed with brine and dried over sodium sulfate. Purification by column chromatography afforded intermediate 9. Yield 52%.

[0159] The obtained compound 9 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were as follows: 1 H NMR (400 MHz, CDC13) δ 7.58 (d, J = 1.6 Hz, 1H), 7.54 (dd, J = 8.4, 1.6 Hz, 1H), 7.18 (d, J = 8.4 Hz, 1H), 6.69 (t, J = 75.2 Hz, 1H), 4.94 (dd, J = 10.4, 8.0 Hz, 1H), 4.68 (t, J = 8.0 Hz, 1H), 4.61 (dd, J = 10.4, 8.0 Hz, 1H), 3.94 (d, J = 6.8 Hz, 2H), 3.84 (s, 3H), 1.32 - 1.29 (m, 1H), 0.68 - 0.60 (m, 2H), 0.37 - 0.32 (m, 2H). ESI-MS m / z: calcd for C 16 H 18 O5NF2 + [M+H] +, 342.0; found, 342.0.

[0160] (4) Synthesis of compound 10:

[0161] Trichlorobromomethane (5.90 mmol, 5.0 equiv.) was added to a solution of compound 9 (1.18 mmol, 1.0 equiv.) and DBU (2.95 mmol, 2.5 equiv.) in DMF, and the reaction mixture was stirred at 0 °C for 20 h. The resulting precipitate was collected by filtration to give compound 10. Yield 65%.

[0162] The resulting compound 10 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were: 1 H NMR (600 MHz, DMSO-d6) δ 8.97 (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). 13 C NMR (151 MHz, DMSO-d6) δ 161.0, 160.6, 150.2, 145.9, 142.1, 133.5, 123.9, 121.2, 119.1, 116.5 (t, J = 256.5 Hz), 111.7, 73.3, 51.9, 9.9, 3.0 (2 x C). ESI-HRMS m / z: calculated for C 16 H 15 O5NF2Na + [M+Na] + , 362.0811; found, 362.0822.

[0163] (5) Synthesis of compound 11:

[0164] Compound 10 (8.84 mmol, 1.0 equiv.) was weighed into 15 mL of a 1:1 mixture of ethanol and water, and 2.12 g of sodium hydroxide (53.04 mmol, 6.0 equiv.) was added. The reaction was stirred at room temperature and monitored by TLC. After the reaction was complete, the ethanol was removed by rotary evaporation, and an appropriate amount of water was added. The pH was adjusted to 6 with dilute hydrochloric acid. The solid was filtered and dried to obtain 2.8 g of yellow solid compound 11, with a yield of 97%.

[0165] The resulting compound 11 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were:1 H NMR (400 MHz, CDC13) δ 8.39 (s, 1H), 7.71 (d, J = 2.0 Hz, 1H), 7.66 (dd, J = 8.4, 2.0 Hz, 1H), 7.26 (d, 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). 13 C NMR (101 MHz, CDC13) δ 165.7, 162.1, 151.0, 145.3, 142.9, 133.8, 124.5, 122.8, 120.0, 115.9 (t, J = 259.3 Hz), 112.6, 74.4, 10.1, 3.4 (2 x C). ESI-MS m / z: Calcd for C 15 H 13 O5NF2 + [M+H] + , 326.1; Found 326.1.

[0166] (6) Synthesis of compounds B1-B12:

[0167] Take 100 mg of compound 11 (0.31 mmol, 1.0 equiv.) dissolved in DMF, then add 117 mg of HATU (0.31 mmol, 1.0 equiv.), amine reagent (0.31 mmol, 1.0 equiv.) and 80 mg of DIPEA (0.62 mmol, 2 equiv.). Stir the reaction at room temperature, monitor by TLC, when there is no starting material, quench with water, extract with ethyl acetate once, wash the organic phase with water three times, wash with saturated sodium chloride solution once, dry the ethyl acetate layer with anhydrous sodium sulfate, concentrate by rotary evaporation, add silica gel to the sample, and purify by column chromatography to obtain the product B1-B10. Add different amine reagents (0.31 mmol, 1.0 equiv.) and thionyl chloride to the DCM solution of intermediate 11 (0.31 mmol, 1.0 equiv.). Stir the reaction at 40°C for 8 hours, then add water, extract the mixture with ethyl acetate, wash the organic layer with brine, and dry with Na2S04. Purify by column chromatography to obtain compound B11 or compound B12.

[0168] The compounds B1-B12 are arranged in the following order:

[0169] Synthesis of compound B1 in Example 20:

[0170] The synthetic procedure was referred to example 19, the amine reagent of the last step reaction was chosen as 2,6-difluorobenzylamine to obtain compound B1, yield: 86%; 1 H NMR (400 MHz, Benzene-d6) δ 8.36 (t, J = 4.8 Hz, 1H), 8.23 - 8.15 (m, 3H), 7.56 - 7.47 (m, 2H), 7.06 (d, J = 8.4 Hz, 1H), 6.48 (t, J = 75.2 Hz, 1H), 6.33 - 6.29 (m, 1H), 5.08 (d, J = 5.2 Hz, 2H), 3.47 (d, J = 6.8 Hz, 2H), 1.46 - 1.35 (m, 1H), 0.45 - 0.38 (m, 2H), 0.20 - 0.12 (m, 2H). 13 C NMR (101 MHz, Benzene-d6) δ 161.7 (dd, J = 248, 8 Hz, 2 x C), 160.7, 160.2, 150.9, 142.6, 141.4, 137.2, 129.9 (t, J = 10.3 Hz), 124.9, 122.9, 119.8, 116.0 (t, J = 259.2 Hz), 113.6 (t, J = 19 Hz), 112.3, 111.6 (dd, J = 18, 6 Hz, 2 x C), 74.2, 31.0, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O4N2F4Na + [M+Na] + 473.1095; found 473.1097.

[0171] Synthesis of compound B2

[0172] The synthetic procedure was referred to example 19, the amine reagent of the last step reaction was chosen as 2-methylaminopyrimidine (CAS number: 75985-45-4) to obtain compound B2, yield: 86%; 1 H NMR (400 MHz, Benzene-d6) δ 8.36 (t, J = 4.8 Hz, 1H), 8.23 - 8.15 (m, 3H), 7.56 - 7.47 (m, 2H), 7.06 (d, J = 8.4 Hz, 1H), 6.48 (t, J = 75.2 Hz, 1H), 6.33 - 6.29 (m, 1H), 5.08 (d, J = 5.2 Hz, 2H), 3.47 (d, J = 6.8 Hz, 2H), 1.46 - 1.35 (m, 1H), 0.45 - 0.38 (m, 2H), 0.20 - 0.12 (m, 2H). 13C NMR (101 MHz, methanol-d4) δ 167.5, 163.2, 162.4, 158.7 (2 x C), 152.2, 144.2, 143.0, 138.4, 126.0, 123.3, 121.2, 120.6, 117.8 (t, J = 255.9 Hz), 113.4, 75.1, 45.8, 11.0, 3.6 (2 x C). ESI-HRMS m / z: calcd for C 20 H 18 O4N4F2Na + [M+Na] + 439.1185.

[0173] Synthesis of compound B3 in Example 22:

[0174] The synthesis procedure was referred to Example 19, and the amine reagent for the last step reaction was chosen as 6-aminobenzothiazole to give compound B3 in yield of 75%; 1 H NMR (400 MHz, CDCl3) δ 8.97 (br, 1H), 8.95 (s, 1H), 8.74 (d, J = 2.4 Hz, 1H), 8.36 (s, 1H), 8.11 (d, J = 8.8 Hz, 1H), 7.68 (dd, J = 8.4, 2.0 Hz, 1H), 7.65 (d, J = 2.0 Hz, 1H), 7.59 (dd, J = 8.8, 2.0 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 6.73 (t, J = 75.2 Hz, 1H), 4.01 (d, J = 7.2 Hz, 2H), 1.40 - 1.34 (m, 1H), 0.74 - 0.67 (m, 2H), 0.45 - 0.40 (m, 2H). 13 C NMR (101 MHz, CDCl3) δ 160.9, 158.6, 153.7, 151.0, 150.3, 142.8, 141.9, 137.5, 135.2, 135.0, 124.7, 123.9, 123.0, 120.0, 119.2, 115.9 (t, J = 259.4 Hz), 112.7, 112.4, 74.3, 10.2, 3.5 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O4N3SF2 + [M+Na] + 458.0970.

[0175] Synthesis of compound B4 in Example 23:

[0176] The synthetic procedure was referred to example 19, the amine reagent of the last step reaction was selected as 2,4-difluorobenzylamine to obtain compound B4, yield: 38%; 1 H NMR (400 MHz, CDC13) δ 8.24 (s, 1H), 7.61-7.56 (m, 2H), 7.45-7.37 (m, 2H), 7.24 (d, J = 8.0 Hz, 1H), 6.88-6.79 (m, 2H), 6.70 (t, J = 74.8 Hz, 1H), 4.6 (d, J = 6.0 Hz, 2H), 4.0 (d, J = 7.2 Hz, 2H), 1.37-1.30 (m, 1H), 0.70-0.65 (m, 2H), 0.41-0.36 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 162.6 (dd, J = 247, 11.9 Hz), 160.8, 160.7, 161.1 (dd, J = 247, 11.9 Hz), 150.9, 142.6, 141.3, 137.2, 131.4 (dd, J = 9.6, 5.8 Hz), 124.8, 122.9, 121.1 (dd, J = 15.0, 3.7 Hz), 119.8, 115.9 (t, J = 259.3 Hz), 112.3, 111.6 (dd, J = 21.0, 3.7 Hz), 104.1 (t, J = 25.4 Hz), 74.2, 36.5, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O4N2F4Na + [M+Na] + 473.1095; found 473.1094.

[0177] Synthesis of compound B5 in example 24:

[0178] The synthetic procedure was referred to example 19, the amine reagent of the last step reaction was selected as 2-aminothiazole to obtain compound B5, yield: 40%; 1 H NMR (500 MHz, DMSO-d6) δ 12.37 (br, 1H), 9.03 (s, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.66 (dd, J = 8.5, 2.0 Hz, 1H), 7.56 (d, J = 3.5 Hz, 1H), 7.38 (d, J = 8.0 Hz, 1H), 7.30 (d, J = 3.5 Hz, 1H), 7.23 (t, J = 74.0 Hz, 1H), 4.02 (d, J = 6.5 Hz, 2H), 1.39-1.23 (m, 1H), 0.66-0.51 (m, 2H), 0.45-0.33 (m, 2H).13 C NMR (151MHz, DMSO-d6) δ 160.3, 158.4, 157.9, 150.1, 143.9, 142.1, 137.6, 135.6, 124.0, 121.2, 119.2, 116.5 (t, J = 256.5Hz), 114.1, 111.9, 73.3, 9.9, 3.1 (2×C). ESI-HRMS m / z: calculated values ​​are for C 18 H 15 O4N3F2SNa + [M+Na] + , 430.0644; the measured value is 430.0656.

[0179] Example 25 Synthesis of compound B6:

[0180] The synthesis steps were as described in Example 19. The amine reagent used in the final reaction was 2,4,6-trifluorobenzylamine, which yielded compound B6 with a yield of 80%. 1 H NMR (400MHz, CDCl3) δ8.23 (s, 1H), 7.60-7.56 (m, 2H), 7.29 (t, J=6.0Hz, 1H), 7.23 (d, J=8.4Hz, 1H), 6.88-6.51 (m, 2H), 6.7 0 (t, J=73.2Hz, 1H), 4.68 (d, J=6.0Hz, 2H), 3.94 (d, J=6.8Hz, 2H), 1.35-1.29 (m, 1H), 0.70-0.65 (m, 2H), 0.40-0.36 (m, 2H). 13 C NMR (101MHz, CDCl3) δ 162.5 (dt, J = 248, 16Hz), 161.9 (ddd, J = 242, 15, 11Hz, 2×C), 160.8, 160.3, 150.9, 142.7, 141.4, 137.1, 124.9, 122.9, 119.9, 116.0 (t, J = 259.3Hz), 112.3, 110.1 (td, J = 19.8, 4.7Hz), 100.6 (ddd, J = 28, 25, 3Hz, 2×C), 74.3, 30.7, 10.2, 3.4 (2×C). ESI-HRMS m / z: calculated values ​​are for C 22 H 17 O4N2F5Na + [M+Na] + , 491.1001; the measured value is 491.1018.

[0181] Example 26 Synthesis of compound B7:

[0182] The synthetic procedure was referred to example 19, and the amine reagent for the last step reaction was chosen as 2-aminobenzothiazole to give compound B7 in 47% yield; 1 H NMR (400 MHz, CDC13) δ 10.33 (br, 1H), 8.42 (s, 1H), 7.88-7.83 (m, 2H), 7.66 (d, J = 2.0 Hz, 1H), 7.61 (dd, J = 8.4, 2.0 Hz, 1H), 7.50-7.46 (m, 1H), 7.37-7.33 (m, 1H), 7.29 (d, J = 8.4 Hz, 1H), 6.74 (t, J = 75.2 Hz, 1H), 4.02 (d, J = 6.8 Hz, 2H), 1.41-1.34 (m, 1H), 0.75-0.70 (m, 2H), 0.48-0.44 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 161.4, 158.3, 157.0, 151.1, 148.7, 142.9, 142.8, 135.7, 132.5, 126.6, 124.4, 124.3, 123.0, 121.7, 121.3, 119.7, 115.9 (t, J = 259.4 Hz), 112.3, 74.4, 10.2, 3.5 (2 x C). ESI-HRMS m / z: calcd for C 22 H 17 04N3F2SNa + [M+Na] + , 480.0800; found 480.0795.

[0183] Synthesis of compound B8 in example 27:

[0184] The synthetic procedure was referred to example 19, and the amine reagent for the last step reaction was chosen as aniline to give compound B8 in 85% yield. 1 H NMR (400 MHz, CDC13) δ 10.33 (br, 1H), 8.42 (s, 1H), 7.88-7.83 (m, 2H), 7.66 (d, J = 2.0 Hz, 1H), 7.61 (dd, J = 8.4, 2.0 Hz, 1H), 7.50-7.46 (m, 1H), 7.37-7.33 (m, 1H), 7.29 (d, J = 8.4 Hz, 1H), 6.74 (t, J = 75.2 Hz, 1H), 4.02 (d, J = 6.8 Hz, 2H), 1.41-1.34 (m, 1H), 0.75-0.70 (m, 2H), 0.48-0.44 (m, 2H). 13C NMR (101 MHz, CDC13) δ 160.8, 158.5, 151.0, 142.8, 141.8, 137.7, 137.5, 129.3 (2 x C), 124.8 (2 x C), 123.0, 120.0 (2 x C), 119.9, 116.0 (t, J = 258.2 Hz), 112.4, 74.3, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 21 H 18 04N2F2Na + [M + Na] + 423.1135.

[0185] Synthesis of compound B9 in Example 28:

[0186] The synthetic procedure was referred to Example 19, and the amine reagent in the last step reaction was chosen as benzylamine to give compound B9 in yield of 68%; 1 H NMR (400 MHz, CDC13) δ 8.26 (s, 1H), 7.61-7.56 (m, 2H), 7.41-7.29 (m, 6H), 7.24 (d, J = 8.4 Hz, 1H), 6.70 (t, J = 75.2 Hz, 1H), 4.66 (d, J = 6.0 Hz, 2H), 3.95 (d, J = 7.2 Hz, 2H), 1.36-1.30 (m, 1H), 0.70-0.64 (m, 2H), 0.41-0.36 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 160.7, 160.6, 150.9, 142.6, 141.3, 138.1, 137.4, 128.9 (2 x C), 128.1 (2 x C), 127.8, 124.9, 122.9, 119.8, 116.0 (t, J = 258.0 Hz), 112.3, 74.2, 43.2, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 22 H 20 04N2F2Na + [M + Na] + 437.1299.

[0187] Synthesis of compound B10 in Example 29:

[0188] The synthetic procedure was referred to Example 19, and the amine reagent in the last step reaction was chosen as 4-aminopyridine to give compound B10 in yield of 100%. 1H NMR (400 MHz, CDC13) δ 8.87 (br, 1H), 8.57 (d, J = 6.4 Hz, 2H), 8.36 (s, 1H), 7.70-7.61 (m, 4H), 7.29 (d, J = 8.0 Hz, 1H), 6.73 (t, J = 74.8 Hz, 1H), 4.01 (d, J = 6.8 Hz, 2H), 1.38-1.32 (m, 1H), 0.74-0.66 (m, 2H), 0.44-0.38 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 161.1, 159.0, 151.0 (2 x C), 144.4, 142.9, 142.4 (2 x C), 137.0, 124.5, 123.0, 120.1, 115.9 (t, J = 259.6 Hz), 113.8 (2 x C), 112.4, 74.4, 10.2, 3.5 (2 x C). ESI-MS m / z: calcd for C 20 H 17 O4N3F2 + [M+Na] + , 424.1; found 424.1.

[0189] Synthesis of compound B11 of example 30:

[0190] Synthesis step refer to example 19, the reagent of last step reaction is N-hydroxysuccinimide, to get compound B11, yield: 85%; 1 H NMR (400 MHz, CDC13) δ 8.50 (s, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.66 (dd, J = 8.0, 2.0 Hz, 1H), 7.31-7.27 (m, 1H), 6.72 (t, J = 74.8 Hz, 1H), 3.97 (d, J = 6.8 Hz, 2H), 2.92 (s, 4H), 1.35-1.31 (m, 1H), 0.70-0.65 (m, 2H), 0.41-0.36 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 168.9 (2 x C), 162.6, 156.5, 151.0, 146.4, 143.1, 130.0, 124.1, 122.9, 120.1, 115.9 (t, J = 258.2 Hz), 112.8, 74.4, 25.8 (2 x C), 10.2, 3.4 (2 x C). ESI-MS m / z: calcd for C 19 H 17 O7N2F2 + [M+H] +Found 423.1; calculated 423.1.

[0191] Example 31 Synthesis of compound B12:

[0192] Synthesis step refer to example 19, the reagent of last step reaction was selected as 1H- benzo[d][1,2,3]triazole, to obtain compound B12, yield: 53%; 1 H NMR (400 MHz, CDC13) δ 9.15 (s, 1H), 8.47 (d, J = 8.4 Hz, 1H), 8.19 (d, J = 8.4 Hz, 1H), 7.81 (d, J = 2.0 Hz, 1H), 7.77-7.72 (m, 2H), 7.61-7.56 (m, 1H), 7.30 (d, J = 8.4 Hz, 1H), 6.74 (t, J = 74.8 Hz, 1H), 4.01 (d, J = 7.2 Hz, 2H), 1.41-1.32 (m, 1H), 0.73-0.66 (m, 2H), 0.43-0.38 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 161.8, 158.1, 151.0, 147.9, 146.0, 143.0, 133.9, 132.0, 131.2, 126.9, 124.3, 122.9, 120.5, 120.2, 116.0 (t, J = 258.2 Hz), 114.9, 112.8, 74.5, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 21 H 16 O4N4F2 + [M+H] + Found 449.1032, calcd 449.1057.

[0193] Example 32 Synthesis of compounds in series C

[0194] (1) Synthesis of compound 12:

[0195] Dissolve 1.0 g of compound 1 (7.24 mmol, 1.0 equiv.) in 10 mL of DMF solvent, then add 0.72 g of sodium hydroxide (18.10 mmol, 2.5 equiv.) and 2.52 g of ethyl difluoro chloroacetate (15.93 mmol, 2.2 equiv.), heat to 80°C for 12h, TLC monitoring. After the reaction is completed, quench the reaction with water, extract with ethyl acetate, dry the ethyl acetate layer with anhydrous sodium sulfate, concentrate the organic phase, and purify by flash column chromatography to obtain 0.93 g of compound 12, yield 54%.

[0196] The obtained compound 12 was identified by nuclear magnetic resonance spectrum, mass spectrometry, and the identification result was: 1 H NMR (400 MHz, CDC13) δ 9.91 (s, 1H), 7.75-7.73 (m, 2H), 7.39 (d, J = 8.8 Hz, 1H), 6.64 (t, J = 72.8 Hz, 1H), 6.59 (t, J = 72.8 Hz, 1H). ESI-MS m / z: calculated C9H7O3F4 + [M+H] + , 239.0; found 239.0.

[0197] (2) Synthesis of compound 13:

[0198] 3,3-dibromo-1,1,1-trifluoro-2-ketone (42.0 mmol, 2.0 equiv.) was added to the aqueous sodium acetate (84.0 mmol, 4.0 equiv.) and stirred at 100°C for 1 hour, after cooling to room temperature, the intermediate 12 (21.0 mmol, 1.0 equiv.) was added to the mixture of methanol and ammonia water. The reaction mixture was stirred at room temperature overnight. Then the methanol was removed by rotary evaporation. Extracted with ethyl acetate for 3 times, washed with saturated sodium chloride solution, and purified by column chromatography to obtain the intermediate 13 with a yield of 64%.

[0199] The obtained compound 13 was identified by nuclear magnetic resonance spectrum, mass spectrometry, and the identification result was: 1 H NMR (500 MHz, CDC13) δ 12.06 (br, 1H), 7.58 (d, J = 2.0 Hz, 1H), 7.53 (dd, J = 8.5, 2.0 Hz, 1H), 7.49 (d, J = 1.0 Hz, 1H), 7.12 (d, J = 8.5 Hz, 1H), 6.48 (t, J = 73.0 Hz, 1H), 6.39 (t, J = 73.0 Hz, 1H). ESI-HRMS m / z: calculated C 12 H8O2N2F7 + [M+H] + , 345.0469; found 345.0429.

[0200] (3) Synthesis of compound 14:

[0201] Intermediate 13 (0.13 mmol, 1.0 equiv.) was dissolved in a mixture of ethanol and water 1:1, sodium hydroxide (2.60 mmol, 20 equiv.) was added, stirred at 80 °C, monitored by TLC. After the reaction was completed, ethanol was removed by rotary evaporation. Column chromatography was used to obtain intermediate 14. Yield: 63%.

[0202] The resulting compound 14 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were: 1 H NMR (400 MHz, DMSO-d6) δ 13.24 (br, 1H), 8.64 (t, J = 6.4 Hz, 1H), 7.98 (s, 1H), 7.94 (dd, J = 8.4, 1.6 Hz, 1H), 7.86 (s, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.28-7.22 (m, 1H), 7.28 (t, J = 73.2 Hz, 1H), 7.25 (t, J = 73.2 Hz, 1H), 7.17-7.07 (m, 2H), 4.48 (d, J = 6.0 Hz, 2H). 12 H9O4N2F4 + [M+H] + , 321.0; found 321.0.

[0203] (4) Synthesis of compounds C1-C3:

[0204] Intermediate 14 (0.16 mmol, 1.0 equiv.) was dissolved in DMF, HATU (0.16 mmol, 1.0 equiv.), amine reagent (0.16 mmol, 1.0 equiv.) and DIPEA (0.32 mmol, 2.0 equiv.) were added. Stirring at room temperature, using TLC for monitoring. The reaction was extracted with ethyl acetate, dried with Na2SO4. Purified by column chromatography to obtain compounds C1-C3.

[0205] Synthesis of compound C1 in Example 33:

[0206] The synthesis step was referred to Example 32, and the amine reagent of the last step reaction was selected as 2,5-difluorobenzylamine to obtain compound C1, yield: 25%; 1 H NMR (400 MHz, DMSO-d6) δ 13.24 (br, 1H), 8.64 (t, J = 6.4 Hz, 1H), 7.98 (s, 1H), 7.94 (dd, J = 8.4, 1.6 Hz, 1H), 7.86 (s, 1H), 7.49 (d, J = 8.4 Hz, 1H), 7.28-7.22 (m, 1H), 7.28 (t, J = 73.2 Hz, 1H), 7.25 (t, J = 73.2 Hz, 1H), 7.17-7.07 (m, 2H), 4.48 (d, J = 6.0 Hz, 2H). 13C NMR (101 MHz, DMSO-d6) δ 162.4, 158.2 (dd, J = 239.6, 2.0 Hz), 156.0 (d, J = 235 Hz), 144.0, 142.1, 141.9, 136.9, 128.9 (dd, J = 17.6, 7.3 Hz), 128.3, 123.3, 121.6, 121.2, 118.3, 116.7 (dd, J = 24, 9 Hz), 116.6 (t, J = 258.6 Hz), 116.4 (t, J = 258.6 Hz), 115.5 (dd, J = 24.7, 4.9 Hz), 115.0 (dd, J = 23.8, 8.6 Hz), 38.3. ESI-HRMS m / z: calcd for C 19 H 13 O3N3F6Na + [M+Na] + , 468.0753; found 468.0755.

[0207] Synthesis of compound C2 of example 34:

[0208] Synthesis step refer to example 32, the amine reagent of last step reaction was selected as 2,4-difluorobenzylamine, to obtain compound C2, yield: 29%; 1 H NMR (500 MHz, DMSO-d6) δ 13.47 (br, 1H), 8.56 (s, 1H), 8.05 (s, 1H), 7.99 (d, J = 8.5 Hz, 1H), 7.80 (s, 1H), 7.47 (d, J = 8.5 Hz, 1H), 7.43 - 7.37 (m, 1H), 7.28 (td, J = 73.0, 2.0 Hz, 2H), 7.20 (td, J = 9.5, 2.5 Hz, 1H), 7.05 (td, J = 8.5, 2.5 Hz, 1H), 4.47 (d, J = 6.0 Hz, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 162.2, 161.3 (dd, J = 243, 12.0 Hz), 159.8 (dd, J = 246, 12.0 Hz), 144.0, 141.9, 136.9, 130.6, 129.7, 128.2, 123.3, 123.0 (d, J = 13.5 Hz), 121.3, 121.2, 118.1, 116.5 (t, J = 258 Hz), 116.4 (t, J = 258 Hz), 111.3 (dd, J = 21.0, 3.0 Hz), 103.5 (t, J = 25.5 Hz), 35.2. ESI-HRMS m / z: calcd for C 19 H 13O3N3F6Na + [M+Na] + Found 468.0755.

[0209] Synthesis of compound C3

[0210] The synthesis was performed according to the procedure described in example 32, using 2-methylaminopyrimidine (CAS Number: 75985-45-4) as the amine reagent to give compound C3 in 30% yield. 1 H NMR (400 MHz, DMSO-d6) δ 13.23 (br, 1H), 8.78 (d, J = 4.8 Hz, 2H), 8.48 (t, J = 5.6 Hz, 1H), 7.99 (s, 1H), 7.95 (d, J = 8.8 Hz, 1H), 7.85 (s, 1H), 7.50 (d, J = 8.8 Hz, 1H), 7.41 (t, J = 4.8 Hz, 1H), 7.29 (t, J = 73.2 Hz, 1H), 7.28 (t, J = 73.2 Hz, 1H), 4.67 (d, J = 5.6 Hz, 2H). 13 C NMR (101 MHz, DMSO-d6) δ 166.9, 162.1, 157.4 (2 x C), 143.9, 142.1, 142.0, 137.1, 128.3, 123.4, 121.3, 121.3, 119.8, 118.1, 116.6 (t, J = 258.3 Hz), 116.4 (t, J = 258.3 Hz), 44.5. ESI-HRMS m / z: calcd for C 17 H 13 O3N5F4Na + [M+Na] + Found 434.0843.

[0211] Synthesis of compounds of series D

[0212] (1) Synthesis of compound 15:

[0213] Intermediate 3 (16.51 mmol, 1.0 equiv.) was added dropwise to DMF. Potassium carbonate was added. Stirring at room temperature for 24 hours. Stirring at -15 °C turned orange yellow, then at -5 °C, add trichlorobromomethane (33.02 mmol, 2.0 equiv.) and DBU (33.02 mmol, 2.0 equiv.), reaction 1-2 hours, quenched with water. The organic layer was washed with brine, dried over sodium sulfate, concentrated. Purification by column chromatography to obtain intermediate 15. Yield 15%.

[0214] The resulting compound 15 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were: 1 H NMR (400 MHz, CDC13) δ 8.17 (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).13C NMR (101 MHz, CDC13) δ 168.1, 162.0, 151.0, 147.8, 142.3, 131.3, 127.7, 122.9, 120.1, 116.0 (t, J = 258.8 Hz), 112.4, 74.4, 52.7, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 16 H 15 O4NF2SNa + [M+Na] + , 378.0582; found, 378.0581.

[0215] (2) Synthesis of compound 16:

[0216] Intermediate 15 (0.28 mmol, 1.0 equiv.) was added to a mixture of sodium hydroxide (1.40 mmol, 5.0 equiv.) in water and ethanol, stirred at room temperature for 10 hours. Add HCl to adjust the pH to 6, collect the precipitate obtained by filtration to obtain compound 16. Yield 96%.

[0217] The resulting compound 16 was identified by nuclear magnetic resonance spectroscopy, mass spectrometry techniques, and the identification results were: 1H NMR (500 MHz, DMSO-d6) δ 8.18 (s, 1H), 7.63 (s, 1H), 7.51 (d, J = 8.5 Hz, 1H), 7.28 (d, J = 8.0 Hz, 1H), 7.18 (t, J = 74.0 Hz, 1H), 4.01 (d, J = 7.0 Hz, 2H), 1.32 - 1.24 (m, 1H), 0.62 - 0.53 (m, 2H), 0.45 - 0.33 (m, 2H), OH (not observed). 13 C NMR (101 MHz, CDC13) δ 167.6, 164.5, 150.7, 148.5, 142.1, 130.7, 127.7, 122.5, 119.7, 115.9 (t, J = 258.8 Hz), 112.0, 74.0, 10.0, 3.2 (2 x C). ESI-MS m / z: calcd 342.1; found 342.1. 15 H 14 O4NF2S + [M+H] + , 342.1; found 342.1.

[0218] (3) Synthesis of compounds D1-D11:

[0219] Weigh 50 mg of compound 16 (0.15 mmol, 1.0 equiv.) dissolved in N, N- dimethylformamide, add 59 mg of HATU (0.15 mmol, 1.0 equiv.) and 60 mg of DIPEA (0.45 mmol, 3.0 equiv.), stir at room temperature for 20 min, add the appropriate amine reagent (0.15 mmol, 1.0 equiv.), react at room temperature for 6 h. Monitor with TLC, and when the raw material is completely reacted, quench with water, extract with ethyl acetate, wash once with saturated sodium chloride solution, dry the ethyl acetate layer with anhydrous sodium sulfate, concentrate the solution, and purify by column chromatography to obtain compounds D1-D11.

[0220] Synthesis of compound D1 in Example 37:

[0221] The synthesis step is referred to Example 36, and the amine reagent is selected as 2,6- difluorobenzylamine to obtain compound D1, yield: 75%; 1H NMR (600 MHz, DMSO-d6) δ 8.85 (t, J = 5.4 Hz, 1H), 8.31 (s, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.62 (dd, J = 8.4, 1.8 Hz, 1H), 7.42 - 7.34 (m, 1H), 7.31 (s, 1H), 7.19 (t, J = 74.4 Hz, 1H), 7.08 - 7.04 (m, 2H), 4.59 (d, J = 5.4 Hz, 2H), 4.00 (d, J = 7.2 Hz, 2H), 1.32 - 1.21 (m, 1H), 0.63 - 0.54 (m, 2H), 0.43 - 0.26 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 166.3, 161.1 (dd, J = 246.0, 9.0 Hz, 2 x C), 160.1, 150.3 (2 x C), 141.7, 130.6, 129.8 (t, J = 10.5 Hz), 124.7, 121.3, 119.4, 116.6 (t, J = 258.0 Hz), 114.2 (t, J = 19.5 Hz), 112.2, 111.5 (dd, J = 21.0, 6.0 Hz, 2 x C), 73.3, 31.0, 10.0, 3.0 (2 x C). ESI-HRMS m / z: [M+Na] calcd for C 22 H 18 O3N2F4SNa + [M+Na] + found 489.0849.

[0222] Synthesis of compound D2

[0223] Synthesis procedure as in example 36, with the amine reagent 2-methylaminopyrimidine (CAS number: 75985-45-4) to give compound D2 in 88% yield; 1 H NMR (600 MHz, DMSO-d6) δ 8.85 (t, J = 5.4 Hz, 1H), 8.31 (s, 1H), 7.68 (d, J = 2.4 Hz, 1H), 7.62 (dd, J = 8.4, 1.8 Hz, 1H), 7.42 - 7.34 (m, 1H), 7.31 (s, 1H), 7.19 (t, J = 74.4 Hz, 1H), 7.08 - 7.04 (m, 2H), 4.59 (d, J = 5.4 Hz, 2H), 4.00 (d, J = 7.2 Hz, 2H), 1.32 - 1.21 (m, 1H), 0.63 - 0.54 (m, 2H), 0.43 - 0.26 (m, 2H). 13C NMR (151 MHz, DMSO-d6) δ 166.7, 166.3, 160.6, 157.4 (2 x C), 150.4, 150.3, 141.7, 130.7, 124.5, 121.4, 119.9, 119.3, 116.6 (t, J = 258.5 Hz), 112.0, 73.3, 44.9, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 20 H 18 O3N4F2SNa + [M+Na] + found 455.0950.

[0224] Synthesis of compound D3 of example 39:

[0225] Synthesis procedure as example 36, amine reagent selected as 2,4-difluorobenzylamine to give compound D3 in 100% yield; 1 H NMR (600 MHz, DMSO-d6) δ 9.10 (t, J = 6.0 Hz, 1H), 8.23 (s, 1H), 7.66 (d, J = 2.4 Hz, 1H), 7.52 (dd, J = 8.4, 1.8 Hz, 1H), 7.35 - 7.29 (m, 1H), 7.23 (d, J = 8.4 Hz, 1H), 7.05 (t, J = 73.2 Hz, 1H), 7.08 - 7.02 (m, 1H), 6.95 (dd, J = 8.4, 2.4 Hz, 1H), 4.46 (d, J = 6.6 Hz, 2H), 3.87 (d, J = 7.2 Hz, 2H), 1.21 - 1.13 (m, 1H), 0.57 - 0.47 (m, 2H), 0.32 - 0.19 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 167.3, 162.0 (dd, J = 243, 12.0 Hz), 161.6, 160.6 (dd, J = 246, 12.0 Hz), 150.8, 150.5, 142.2, 131.3 (dd, J = 9.0, 6.0 Hz), 131.1, 125.4, 122.6 (dd, J = 15.0, 4.5 Hz), 121.9, 120.1, 117.0 (t, J = 258.0 Hz), 112.4, 111.9 (dd, J = 24.0, 3.0 Hz), 104.2 (t, J = 25.5 Hz), 74.0, 36.4, 10.5, 3.6 (2 x C). ESI-HRMS m / z: calcd for C 22 H 19 O3N2F4S + [M+H]+ 467.1047; found 467.1033.

[0226] Synthesis of compound D4:

[0227] Synthesis procedure as in example 36, amine reagent was selected as 2-aminothiazole to give compound D4 in 81% yield; 1 H NMR (600 MHz, DMSO-d6) δ 8.57 (s, 1H), 7.84 (d, J = 2.4 Hz, 1H), 7.63 (dd, J = 8.4, 1.8 Hz, 1H), 7.56 (d, J = 3.6 Hz, 1H), 7.30 (d, J = 3.6 Hz, 1H), 7.29 (s, 1H), 7.12 (t, J = 74.4 Hz, 1H), 4.00 (d, J = 6.6 Hz, 2H), 1.29 - 1.22 (m, 1H), 0.61 - 0.54 (m, 2H), 0.46 - 0.30 (m, 2H), NH (not observed). 13 C NMR (151 MHz, DMSO-d6) δ 167.5, 159.3, 158.2, 150.8, 148.5, 142.2, 138.3, 130.9, 127.8, 121.8, 120.2, 117.0 (t, J = 258.7 Hz), 115.0, 112.6, 73.9, 10.4, 3.6 (2 x C). ESI-HRMS m / z: calcd for C 18 H 16 O3N3F2S2 + [M+H] + 424.0596; found 424.0585.

[0228] Synthesis of compound D5:

[0229] Synthesis procedure as in example 36, amine reagent was selected as 2,4,6-trifluorobenzylamine to give compound D5 in 100% yield; 1 H NMR (600 MHz, DMSO-d6) δ 8.89 (t, J = 5.4 Hz, 1H), 8.30 (s, 1H), 7.68 (s, 1H), 7.62 (dd, J = 8.4, 1.8 Hz, 1H), 7.31 (d, J = 3.6 Hz, 1H), 7.19 - 7.14 (m, 2H), 7.18 (t, J = 74.4 Hz, 1H), 4.53 (d, J = 6.0 Hz, 2H), 4.01 (d, J = 7.2 Hz, 2H), 1.35 - 1.17 (m, 1H), 0.68 - 0.49 (m, 2H), 0.43 - 0.34 (m, 2H).13 C NMR (151 MHz, DMSO-d6) δ 166.3, 161.4 (dt, J = 246, 16.5 Hz), 161.3 (ddd, J = 248, 34.5, 30.0 Hz, 2 x C), 160.2, 150.3, 150.2, 141.7, 130.7, 124.7, 121.4, 119.4, 116.6 (t, J = 258.0 Hz), 112.3, 111.1 (td, J = 19.5, 4.5 Hz), 100.5 (dd, J = 30.0, 25.5 Hz, 2 x C), 73.4, 30.8, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O3N2F5S + [M+H] + , 485.0953; found 485.0933.

[0230] Synthesis of compound D6 of example 42:

[0231] Synthesis step refer to example 36, amine reagent is selected as 2-aminobenzothiazole, to obtain compound D6, yield: 67%; 1 H NMR (600 MHz, DMSO-d6) δ 12.55 (s, 1H), 8.74 (s, 1H), 8.05 (d, J = 7.8 Hz, 1H), 7.93 (d, J = 1.8 Hz, 1H), 7.82 (d, J = 7.8 Hz, 1H), 7.73 (dd, J = 8.4, 1.8 Hz, 1H), 7.55 - 7.46 (m, 1H), 7.38 - 7.31 (m, 2H), 7.22 (t, J = 74.4 Hz, 1H), 4.08 (d, J = 6.6 Hz, 2H), 1.35 - 1.27 (m, 1H), 0.66 - 0.57 (m, 2H), 0.48 - 0.32 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 166.8, 159.6, 157.9, 150.3, 148.5, 148.0, 141.9, 131.7, 130.5, 128.0, 126.3, 123.9, 121.8, 121.2, 120.5, 119.7, 116.6 (t, J = 258.0 Hz), 112.3, 73.4, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O3N3F2S2 + [M+H] + , 474.0752; found 474.0753.

[0232] Synthesis of compound D7:

[0233] The synthesis was performed according to the procedure described in example 36, using 4-aminopyridine as amine reagent to give compound D7 in 86% yield; 1 H NMR (500 MHz, DMSO-d6) δ 10.55 (s, 1H), 8.58 (s, 1H), 8.51 (d, J = 6.5 Hz, 2H), 7.89 (d, J = 6.5 Hz, 2H), 7.82 (d, J = 2.0 Hz, 1H), 7.71 (dd, J = 8.0, 2.0 Hz, 1H), 7.35 (d, J = 8.5 Hz, 1H), 7.22 (t, J = 74.0 Hz, 1H), 4.07 (d, J = 7.0 Hz, 2H), 1.35 - 1.25 (m, 1H), 0.65 - 0.56 (m, 2H), 0.46 - 0.32 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 166.6, 159.8, 150.3, 150.3 (2 x C), 149.6, 145.2, 141.9, 130.4, 126.8, 121.3, 119.7, 116.6 (t, J = 258.5 Hz), 114.3 (2 x C), 112.4, 73.4, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 20 H 18 O3N3F2S + [M+H] + , 418.1031; found 418, 1023.

[0234] Synthesis of compound D8:

[0235] The synthesis was performed according to the procedure described in example 36, using 2,3-difluorobenzylamine as amine reagent to give compound D8 in 100% yield; 1 H NMR (500 MHz, DMSO-d6) δ 9.14 (t, J = 6.5 Hz, 1H), 8.35 (s, 1H), 7.73 (d, J = 2.0 Hz, 1H), 7.64 (dd, J = 8.5, 2.5 Hz, 1H), 7.36 - 7.25 (m, 2H), 7.22 - 7.15 (m, 2H), 7.19 (t, J = 74.5 Hz, 1H), 4.60 (d, J = 6.0 Hz, 2H), 4.02 (d, J = 7.0 Hz, 2H), 1.33 - 1.24 (m, 1H), 0.63 - 0.56 (m, 2H), 0.40 - 0.36 (m, 2H). 13C NMR (151 MHz, DMSO-d6) δ 166.3, 160.7, 150.3, 150.2, 149.6 (dd, J = 244.5, 13.5 Hz), 147.6 (dd, J = 244.5, 13.5 Hz), 141.7, 130.6, 128.8 (d, J = 12.0 Hz), 124.8, 124.6 (dd, J = 7.5, 4.5 Hz), 124.5 (t, J = 3.0 Hz), 121.3, 119.4, 116.6 (t, J = 258.0 Hz), 115.9 (d, J = 16.5 Hz), 112.1, 73.3, 35.8, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 22 H 18 O3N2F4SNa + [M+Na] + , 489.0866; found, 489.0853.

[0236] Synthesis of compound D9 of example 45:

[0237] Synthesis step refers to example 36, amine reagent is selected as 2-(aminomethyl)naphthalene (CAS No.: 2018-90-8), to obtain compound D9, yield: 100%; 1 H NMR (500 MHz, DMSO-d6) δ 9.21 (t, J = 6.5 Hz, 1H), 8.36 (s, 1H), 7.91-7.85 (m, 3H), 7.82 (s, 1H), 7.75 (d, J = 2.0 Hz, 1H), 7.64 (dd, J = 8.5, 2.0 Hz, 1H), 7.53 (dd, J = 8.5, 1.5 Hz, 1H), 7.50-7.46 (m, 2H), 7.32 (d, J = 8.5 Hz, 1H), 7.19 (t, J = 74.5 Hz, 1H), 4.70 (d, J = 6.5 Hz, 2H), 4.02 (d, J = 7.0 Hz, 2H), 1.32-1.24 (m, 1H), 0.64-0.55 (m, 2H), 0.42-0.29 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 166.3, 160.6, 150.5, 150.3, 141.7, 137.1, 132.9, 132.1, 130.7, 127.9, 127.5, 127.5, 126.2, 126.0, 125.6, 125.4, 124.6, 121.3, 119.4, 116.6 (t, J = 258.3 Hz), 112.1, 73.3, 42.5, 10.0, 3.0 (2 x C). ESI-HRMS m / z: calcd for C26 H 22 O3N2F2SNa + [M+Na] + , 503.1211; found 503.1191.

[0238] Synthesis of compound D10 in Example 46:

[0239] The synthetic procedure was referred to Example 36, and the amine reagent was selected as benzylamine to give compound D10 in 49% yield; 1 H NMR (500 MHz, DMSO-d6) δ 9.10 (t, J = 6.5 Hz, 1H), 8.33 (s, 1H), 7.74 (d, J = 2.0 Hz, 1H), 7.63 (dd, J = 8.0, 2.0 Hz, 1H), 7.36 - 7.31 (m, 5H), 7.27 - 7.22 (m, 1H), 7.19 (t, J = 74.0 Hz, 1H), 4.53 (d, J = 6.5 Hz, 2H), 4.02 (d, J = 7.0 Hz, 2H), 1.32 - 1.24 (m, 1H), 0.65 - 0.54 (m, 2H), 0.43 - 0.30 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 167.3, 161.1, 151.0, 150.8, 142.3, 138.3, 131.3, 128.9 (2 x C), 128.0 (2 x C), 127.7, 123.7, 123.1, 119.9, 116.0 (t, J = 259.1 Hz), 112.2, 74.3, 43.5, 10.2, 3.4 (2 x C). ESI-HRMS m / z: calcd for C 22 H 21 O3N2F2S + [M+H] + , 431.1235; found 431.1229.

[0240] Synthesis of compound D11 in Example 47:

[0241] The synthetic procedure was referred to Example 36, and the amine reagent was selected as aniline to give compound D11 in 46% yield; 1H NMR (600 MHz, DMSO-d6) δ 10.22 (s, 1H), 8.48 (s, 1H), 7.87-7.82 (m, 2H), 7.71 (dd, J = 8.4, 1.8 Hz, 1H), 7.42-7.36 (m, 2H), 7.33 (d, J = 2.4 Hz, 1H), 7.36-7.06 (m, 2H), 7.15 (tt, J = 7.2, 1.2 Hz, 1H), 4.07 (d, J = 6.6 Hz, 2H), 1.35-1.26 (m, 1H), 0.67-0.58 (m, 2H), 0.49-0.36 (m, 2H). 13 C NMR (151 MHz, DMSO-d6) δ 166.4, 159.0, 150.4, 150.3, 141.8, 138.3, 130.6, 128.7 (2 x C), 125.6, 124.1, 121.3, 120.7 (2 x C), 119.6, 116.6 (t, J = 256.5 Hz), 112.4, 73.4, 10.0, 3.1 (2 x C). ESI-HRMS m / z: calcd for C 21 H 19 O3N2F2S + [M+H] + , 417.1079; found 417.1058.

[0242] Example 48 Synthesis of compounds of series E

[0243] (1) Synthesis of compound 18:

[0244] Take 200 mg of raw material 17 (1.10 mmol, 1.0 equiv.), add 116.2 mg of sodium carbonate (1.10 mmol, 1.0 equiv.), add 2.5 mL of reagent N, N-dimethylformamide, and finally add 174 mg of ethyl difluoro chloroacetate (1.10 mmol, 1.0 equiv.), react for 12 hours at a temperature of 80°C, TLC detects that the raw material is completely reacted, add water solution to quench the reaction. Extract 3 times with ethyl acetate, wash the organic phase with saturated sodium chloride solution. Concentrate the organic phase and purify by flash column chromatography to obtain 60 mg of product, with a yield of 24%.

[0245] The obtained compound 18 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows: 1 H NMR (500 MHz, CDCl3) δ 9.92 (s, 1H), 7.16 (s, 2H), 6.66 (t, J = 76.0 Hz, 1H), 3.96 (s, 6H). ESI-MS m / z: calcd for C10 H 11 O2F2 + [M+H] + , 233.1 ; found 233.1.

[0246] (2) Synthesis of compound 19:

[0247] Weigh 1.76 g of 3,3-dibromo-1,1,1-trifluoro-2-ketone (8.6 mmol, 2.0 equiv.) and 1.42 g of sodium acetate (17.2 mmol, 4.0 equiv.) into a reaction bottle, dissolve in a proper amount of water, stir at 100°C for 1 hour, cool to room temperature, then add 1 g of compound 18 (4.3 mmol, 1.0 equiv.) in methanol solution and 4 mL of ammonia water into the above solution, stir at room temperature, when TLC detection shows that the raw material no longer decreases, remove the methanol by rotary evaporation. Add water, extract with ethyl acetate for 3 times, wash the organic phase with saturated sodium chloride solution once, dry the organic phase with anhydrous sodium sulfate. Concentrate the organic phase and purify by flash column chromatography to obtain 890 mg of yellow solid product (compound 19) with a yield of 61%.

[0248] The obtained compound 19 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1 H NMR (500 MHz, DMSO-d6) δ 13.26 (br, 1H), 7.98 (s, 1H), 7.36 (s, 2H), 6.89 (t, J = 75.5 Hz, 1H), 3.89 (s, 6H). ESI-MS m / z: calculated value for C 13 H 12 02N2F5 + [M+H] + , 339.1 ; found 339.1.

[0249] (3) Synthesis of compound 20:

[0250] Weigh 1.6 g of compound 19 (4.73 mmol, 1.0 equiv.) into a mixture of ethanol and water (1:1), add 3.8 g of sodium hydroxide (94.6 mmol, 20 equiv.), stir the reaction at 80°C, monitor by TLC, after the reaction is complete, remove the ethanol by rotary evaporation, add a proper amount of water, adjust the pH to 6 with dilute hydrochloric acid. Filter to obtain a solid, and dry to obtain 778 mg of yellow solid product with a yield of 52%.

[0251] The obtained compound 20 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1H NMR (500 MHz, DMSO-d6) δ 7.88 (s, 1H), 7.48 (s, 2H), 6.90 (t, J = 75.0 Hz, 1H), 3.89 (s, 6H). OH (1H, not observed), NH (1H, not observed). ESI-MS m / z: calcd for C 13 H 13 O5N2F2 + [M+H] + , 315.0; found 315.0.

[0252] (4) Synthesis of compounds E1-E3:

[0253] Take 50 mg of compound 20 (0.09 mmol, 1.0 equiv.) dissolved in DMF, then add 53 mg of HATU (0.09 mmol, 1.0 equiv.), 15 mg of amine reagent (0.1 mmol, 1.1 equiv.) and 36.5 mg of DIPEA (0.27 mmol, 3.0 equiv.). Stir the reaction at room temperature, monitor by TLC, quench with water, extract with ethyl acetate, wash the organic phase with water three times, and saturated sodium chloride solution once, dry the ethyl acetate layer over anhydrous sodium sulfate, concentrate, add silica gel to the sample, and purify by column chromatography to obtain the product E1-E3.

[0254] Example 49 Synthesis of compound E1:

[0255] Synthesis step according to Example 48, with the amine reagent selected as 2- methylaminopyrimidine (CAS number: 75985-45-4) to obtain compound E1, yield: 19%; 1 H NMR (500 MHz, DMSO-d6) δ 8.77 (d, J = 4.5 Hz, 2H), 7.77 (s, 1H), 7.54 (s, 2H), 7.40 (t, J = 5.0 Hz, 1H), 6.88 (t, J = 75.0 Hz, 1H), 4.67 (d, J = 6.0 Hz, 2H), 3.90 (s, 6H), NH (2H, not observed). 13 C NMR (101 MHz, DMSO-d6) δ 167.1, 162.3, 157.4 (2 x C), 152.8 (2 x C), 145.0, 137.0, 128.7, 128.3, 121.1, 119.8, 117.4 (t, J = 258.2 Hz), 102.4 (2 x C), 56.3 (2 x C), 44.5. ESI-HRMS m / z: calcd for C 18 H17 O4N5F2Na + [M+Na] + , 428.1141 ; found 428.1141.

[0256] Synthesis of compound E2 of Example 50:

[0257] The synthesis procedure was referred to Example 48, and the amine reagent was selected as 6-aminobenzothiazole to give compound E2 in 9% yield; 1 H NMR (600 MHz, DMSO-d6 ) δ 13.26 (br, 1H), 10.10 (s, 1H), 9.28 (s, 1H), 8.72 (s, 1H), 8.11-8.02 (m, 2H), 7.99-7.88 (m, 1H), 7.52 (s, 2H), 6.91 (t, J = 75.0 Hz, 1H), 3.92 (s, 6H). 13 C NMR (151 MHz, DMSO-d6) δ 160.9, 154.6, 152.8 (2 x C), 149.2, 145.3, 136.7 (2 x C), 134.1, 128.8, 128.1, 122.8, 122.3, 119.7, 117.4 (t, J = 256.5 Hz), 112.4, 102.7 (2 x C), 56.4 (2 x C). ESI-HRMS m / z: calcd for C 20 H 16 O4N4SF2Na + [M+Na] + , 469.0753; found 469.0737.

[0258] Synthesis of compound E3 of Example 51:

[0259] The synthesis procedure was referred to Example 48, and the amine reagent was selected as 2,6-difluorobenzylamine to give compound E3 in 11% yield; 1 H NMR (500 MHz, DMSO-d6) δ 13.04 (br, 1H), 8.26 (t, J = 246 Hz, 1H), 7.84 (t, J = 211 Hz, 1H), 7.38 (s, 3H), 7.13-7.02 (m, 2H), 6.87 (t, J = 75.5 Hz, 1H), 4.54 (d, J = 5.5 Hz, 2H), 3.86 (s, 6H). 13C NMR (151 MHz, DMSO-d6) δ 161.7, 161.1 (dd, J = 246.0, 7.5 Hz, 2 x C), 152.7 (2 x C), 145.0, 136.8, 129.7, 128.7, 128.2, 121.1, 117.3 (t, J = 258.0 Hz), 117.3, 114.6, 111.5 (dd, J = 21, 6.0 Hz, 2 x C), 102.4, 56.3 (2 x C), 30.3. ESI-HRMS m / z: calcd for C 20 H 17 O4N3F4Na + [M+Na] + , 462.1047; found, 462.1046.

[0260] Synthesis of compounds of Example 52F series

[0261] (1) Synthesis of compound 22:

[0262] Take 200 mg of 3,4,5-trihydroxybenzaldehyde (1.29 mmol, 1.0 equiv.), add 195 mg of sodium carbonate (1.29 mmol, 1.0 equiv.), add 2.5 mL of solvent N,N-dimethylformamide, and finally add 205.7 mg of ethyl difluorochloroacetate (1.29 mmol, 1.0 equiv.). After reacting at a temperature of 70 °C for 12 hours, TLC detects that the raw material is completely reacted, and water is added to quench the reaction. Extract with ethyl acetate 3 times, and wash the organic phase with saturated sodium chloride solution. Concentrate the organic phase and purify by flash column chromatography to obtain 58 mg of product as a white solid, with a yield of 22%.

[0263] The obtained compound 22 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results are as follows: 1 H NMR (500MHz, DMSO-d6) δ 10.33 (br, 2H), 9.77 (s, 1H), 6.96 (t, J = 75.5 Hz, 1H), 6.93 (s, 2H). ESI-MS m / z: calcd for C8H7O4F2 + [M+H] + , 205.0; found, 205.0.

[0264] (2) Synthesis of compound 23:

[0265] Take 180 mg of compound 22 (0.88 mmol, 1.0 equiv.), add 305 mg of potassium carbonate (2.2 mmol, 2.5 equiv.), add 10 mL of solvent N, N-dimethylformamide, and finally add 261.9 mg of bromomethylcyclopropane (1.94 mmol, 2.2 equiv.), react at a temperature of 80°C for 5 hours, TLC detection shows that the raw material is completely reacted, and water is added to quench the reaction. Extract with ethyl acetate 3 times, and wash the organic phase with saturated sodium chloride solution. Concentrate the organic phase and purify by flash column chromatography to obtain 169 mg of product (compound 23) with a yield of 62%.

[0266] The obtained compound 23 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1 H NMR (400MHz, CDCI3) δ 9.86 (s, 1H), 7.10 (s, 2H), 6.74 (t, J = 74.4 Hz, 1H), 3.95 (d, J = 6.8 Hz, 4H), 1.34-1.30 (m, 2H), 0.68-0.64 (m, 4H), 0.40-0.36 (m, 4H). ESI-MS m / z: calculated value of C 16 H 19 O4F2 + [M+H] + , 313.1; found 313.1.

[0267] (3) Synthesis of compound 24:

[0268] Weigh 1.3 g of 3,3-dibromo-1,1,1-trifluoro-2-ketone (4.8 mmol, 1.5 equiv.) and 1.1 g of sodium acetate (12.8 mmol, 4.0 equiv.) into a reaction bottle, dissolve in water, stir at 100°C for 1 hour, cool to room temperature, then add a solution of 1 g of compound 23 (3.2 mmol, 1.0 equiv.) in methanol and 4 mL of ammonia water to the above solution, stir at room temperature, TLC detection, and remove the methanol by rotary evaporation. Add water, extract with ethyl acetate 3 times, wash the organic phase with saturated sodium chloride solution once, and dry the organic phase over anhydrous sodium sulfate. Concentrate the organic phase and purify by flash column chromatography to obtain 1.0 g of yellow solid product with a yield of 74%.

[0269] The obtained compound 24 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification result is: 1H NMR (400MHz, DMSO-d6) δ 13.21 (br, 1H), 7.98 (s, 1H), 7.30 (s, 2H), 6.90 (t, J = 75.2 Hz, 1H), 3.95 (d, J = 6.8 Hz, 4H), 1.31-1.22 (m, 2H), 0.61-0.55 (m, 4H), 0.39-0.34 (m, 4H). ESI-MS m / z: calcd for C 19 H 20 O3N2F5 + [M+H] + , 419.3; found 419.3.

[0270] (4) Synthesis of compound 25:

[0271] Compound 24 (63 mg, 0.27 mmol, 1.0 equiv.) was dissolved in a mixture of ethanol and water (1:1) and sodium hydroxide (21.7 mg, 0.54 mmol, 2.0 equiv.) was added. The reaction was stirred at 80 °C and monitored by TLC. After the reaction was completed, ethanol was removed by rotary evaporation, and water was added. The pH was adjusted to 6 with dilute hydrochloric acid. The solid was filtered and dried to give 42 mg, with a yield of 39%.

[0272] The obtained compound 25 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were: 1 H NMR (400MHz, DMSO-d6) δ 13.21 (br, 1H), 7.98 (s, 1H), 7.30 (s, 2H), 6.90 (t, J = 75.2 Hz, 1H), 3.95 (d, J = 6.8 Hz, 4H), 1.31-1.22 (m, 2H), 0.61-0.55 (m, 4H), 0.39-0.34 (m, 4H). ESI-MS m / z: calcd for C 19 H 21 O5N2F2 + [M+H] + , 395.1; found 395.1.

[0273] (5) Synthesis of compound F1:

[0274] To 50 mg of compound 25 (0.13 mmol, 1.0 equiv.) dissolved in DMF, 50 mg of HATU (0.13 mmol, 1.0 equiv.), amine reagent (0.14 mmol, 1.1 equiv.) and 50 mg of DIPEA (0.39 mmol, 3.0 equiv.) were added. The reaction was stirred at room temperature, monitored by TLC, quenched with water when starting material was consumed, extracted with ethyl acetate once, washed with water three times, washed with saturated sodium chloride solution once, dried over anhydrous sodium sulfate, concentrated, added silica gel to the sample, and purified by column chromatography to give the product as a yellow solid.

[0275] Synthesis of compound F1

[0276] The synthesis was performed according to the procedure described in example 52, using 2-methylaminopyrimidine (CAS number: 75985-45-4) as amine reagent to give compound F1 in 8% yield; 1 H NMR (500 MHz, DMSO-d6) δ 13.53 (br, 1H), 8.76 (d, J = 4.5 Hz, 2H), 8.52 (s, 1H), 7.78 (s, 1H), 7.50 (s, 2H), 7.39 (t, J = 4.5 Hz, 1H), 6.89 (t, J = 75.5 Hz, 1H), 4.66 (d, J = 6.0 Hz, 2H), 3.97 (d, J = 7.0 Hz, 4H), 1.30 - 1.23 (m, 2H), 0.61 - 0.55 (m, 4H), 0.38 - 0.35 (m, 4H). 13 C NMR (101 MHz, DMSO-d6) δ 167.1, 162.4, 157.4 (2 x C), 152.2 (2 x C), 145.2, 136.8, 129.3, 128.2, 120.9, 119.9 (2 x C), 117.5 (t, J = 256.4 Hz), 103.7, 73.3 (2 x C), 44.5, 10.1 (2 x C), 3.1 (4 x C). ESI-HRMS m / z: calcd for C 24 H 25 O4N5F2Na + [M+Na] + 508.1767; found 508.1760.

[0277] Synthesis of compounds of series G

[0278] (1) Synthesis of compound 27:

[0279] Weigh 200 mg of 3,3-dibromo-1,1,1 -trifluoro-2-ketone (0.74 mmol, 1.12 equiv.) and 193 mg of sodium acetate (1.48 mmol, 2.24 equiv.) into a reaction flask, dissolve in a suitable amount of water, stir at 100°C for 1 hour, cool to room temperature, then add a solution of 88.8 mg of compound 26 (0.66 mmol, 1.0 equiv.) in methanol and 4 mL of ammonia water to the above solution, stir at room temperature, monitor by TLC, and remove the methanol by rotary evaporation. Add water, extract three times with ethyl acetate, wash the organic phase once with saturated sodium chloride solution, and dry the organic phase over anhydrous sodium sulfate. Concentrate the organic phase and purify by flash column chromatography to obtain 83 mg of yellow solid product with a yield of 52%.

[0280] The obtained compound 27 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1 H), 7.84-7.75 (m, 2H), 7.22 (d, J = 8.0 Hz, 1 H), 2.26 (s, 3H), 2.24 (s, 3H), OH (1 H, not observed). NH (1 H, not observed). ESI-MS m / z: calculated value C 12 H 12 N2F3 + [M+H] + , 241.0947; found 241.0908.

[0281] (2) Synthesis of compound 28:

[0282] Weigh 39 mg of compound 27 (0.16 mmol, 1.0 equiv.) into a mixture of ethanol and water (1 :1 ), add 8.9 mg of sodium hydroxide (0.23 mmol, 1.4 equiv.), stir the reaction at 80°C, monitor by TLC, and after the reaction is complete, remove the ethanol by rotary evaporation, add a suitable amount of water, and adjust the pH to 6 with dilute hydrochloric acid. Filter to obtain a solid, and dry to obtain 30 mg with a yield of 86%.

[0283] The obtained compound 28 was identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results were as follows: 1 H NMR (400 MHz, DMSO-d6) δ 7.89 (s, 1 H), 7.84-7.75 (m, 2H), 7.22 (d, J = 8.0 Hz, 1 H), 2.26 (s, 3H), 2.24 (s, 3H), OH (1 H, not observed). NH (1 H, not observed). ESI-MS m / z: calculated value C12 H 12 O2N2 + [M+H] + , 217.0; found 217.0.

[0284] (3) Synthesis of compound G1:

[0285] Take 100 mg of compound 28 (0.31 mmol, 1.0 equiv.) dissolved in DMF, then add 117 mg of HATU (0.31 mmol, 1.0 equiv.), 44 mg of 4-aminopyridine (0.31 mmol, 1.0 equiv.) and 80 mg of DIPEA (0.62 mmol, 2.0 equiv.). Stir the reaction at room temperature, monitor by TLC, when the starting material is consumed, quench with water, extract with ethyl acetate three times, wash with saturated sodium chloride solution once, dry the ethyl acetate layer over anhydrous sodium sulfate, concentrate by rotary evaporation, add silica gel to the sample, purify by column chromatography to obtain 78 mg of yellow solid product G1, with a yield of 86%.

[0286] The obtained compound G1 is identified by nuclear magnetic resonance spectroscopy and mass spectrometry, and the identification results are as follows: 1 H NMR (500 MHz, DMSO-d6) δ 13.41 (br, 1H), 10.23 (br, 1H), 8.46 (d, J = 5.5 Hz, 2H), 8.00 (s, 1H), 7.94 (s, 1H), 7.91 (d, J = 5.5 Hz, 2H), 7.85 (d, J = 7.5 Hz, 1H), 7.24 (d, J = 7.5 Hz, 1H), 2.29 (s, 3H), 2.26 (s, 3H). ESI-HRMS m / z: calculated C 17 H 17 ON4 + [M+H] + , 293.1397; found 293.1390.

[0287] Active examples

[0288] Enzyme activity test of active example 1

[0289] The compounds of the present application are subjected to biological tests by the following method:

[0290] 1. Preparation of reaction buffer and reaction termination solution (reagents are shown in Table 1)

[0291] (1) Preparation of 1-fold reaction buffer

[0292] IMAP reaction buffer containing 0.1% BSA (5x) (provided with IMAP FP IPP Explorer Kit) was diluted into 1x reaction buffer with 1 mM DTT.

[0293] (2) Preparation of reaction stop solution

[0294] IMAP process binding buffer A (5x), IMAP process binding buffer B (5x), and IMAP process binding reagent (provided with IMAP FP IPP Explorer Kit) were prepared according to the instruction manual to prepare the reaction stop solution.

[0295] 2. Compound preparation

[0296] (1) Compound dilution

[0297] A solution of 100 times the final concentration of the compound was prepared. The compound was diluted in gradient to the set number of concentration points using an automated microplate pipette (Precision PRC384U) as follows: if 5 times dilution was performed, 50 μL of the starting concentration of the compound DMSO solution was added to well A2 in the Echo 384 well plate, and 40 μL of 100% DMSO was added to wells A3-A11; 10 μL of the compound from well A2 was added to well A3, mixed, and 5 times dilution was performed in sequence to obtain 10 concentration points; 40 μL of 100% DMSO was added to wells Al and A12.

[0298] (2) Transfer of compound to 384 reaction plate

[0299] 200 nL of the compound was transferred from the above diluted Echo 384 well plate to a 384 well reaction plate using an Echo 550 instrument, and 200 nL of 100% DMSO was transferred to the negative control and the positive control.

[0300] 3. Enzymatic reaction

[0301] (1) Preparation of 2 times enzyme solution

[0302] PDE4D was added to 1 times reaction buffer to form a 2 times enzyme solution (PDE4D final concentration: 0.00625 μg / ml).

[0303] (2) Preparation of 2 times substrate solution

[0304] For enzyme PDE4D, add FAM-labeled cAMP to 1x reaction buffer to make a 2x substrate solution (FAM-cAMP final concentration: 0.1 μM).

[0305] (3) Add enzyme solution to 384-well plate

[0306] Add 10 μL of 2x enzyme solution to each well of the 384-well reaction plate. For enzyme- free control wells, replace enzyme solution with 10 μL of 1x reaction buffer. Centrifuge at 1000 rpm for 1 min and incubate at room temperature for 15 min.

[0307] (4) Add substrate solution to 384-well plate to start enzymatic reaction

[0308] Add 10 μL of 2x substrate solution to each well of the 384-well reaction plate. Centrifuge at 1000 rpm for 1 min. Incubate at 25°C for 30 min.

[0309] (5) Termination of enzyme reaction

[0310] Add 60 μL of reaction termination solution to each well of the 384-well reaction plate to stop the reaction. Incubate at room temperature for 60 min with shaking at 600 rpm in the dark.

[0311] 4. Read data with EnVision and calculate data

[0312] Read data with EnVision.

[0313] 5. Calculate inhibition and IC 50 Curve fitting

[0314] Copy data from EnVision, where the maximum value is the DMSO control reading and the minimum value is the enzyme-free control reading. Inhibition (%) = (maximum value - sample value) / (maximum value - minimum value) x 100%.

[0315] Import data into MS Excel and fit IC values with XLFit excel add-in version 5.4.0.8 50 ;

[0316] Fit equation: Y = Bottom + (Top - Bottom) / (1 + (IC 50 / X)A HillSlope)

[0317] Table 1 Reagent information

[0318] The PDE4B, PDE4D enzyme inhibitory effect of the compounds provided by the examples was determined according to the above method, and the results are shown in Table 2, which is the determination results of the PDE4D enzyme inhibitory effect of the compounds of the present application.

[0319] Table 2 Inhibition rate of compounds on PDE4D

[0320] The IC of the active example 2 compound on PDE4B, PDE4D 50 detection

[0321] The IC value was obtained by referring to the active example 1. 50

[0322] The IC value of the compound is shown in Table 3. 50

[0323] Table 3 IC value of compounds 50

[0324] Inhibition of RAW 264.7 cell inflammation by the compounds of the present application in active example 3

[0325] The cell inflammatory factor expression level inhibition test of the compounds of the present application was carried out by using the conventional ELISA method.

[0326] 1. Cell culture

[0327] The mouse monocyte macrophage Raw 264.7 was cultured in DMEM high-sugar medium added with 10% (V / V) FBS, 100 μg / mL penicillin and 100 μg / mL streptomycin. The culture condition was 37℃, 5% CO2, and the cells were subcultured when they reached 80% confluence.

[0328] 2. Cell administration and cell inflammation induction

[0329] The cells in the logarithmic growth phase were used for the experiment, and the cells were inoculated in a 12-well plate at 2*10 5 / cm2, and were cultured at 37℃, 5% CO2 until they grew to 70%, and were ready for use. The groups were set: blank group, LPS stimulation group, positive control group (Roflumilast) and administration group (compound of the present application). The culture medium was carefully removed, and fresh complete medium containing the compound was added to the positive control group and the administration group, and the same volume of DMSO was added to the blank group and the LPS stimulation group. After 1 hour, 1 μg / mL LPS was added to each group except the blank group to induce cell inflammation for 4 hours. The well plate was taken out, and the ELISA experiment was carried out according to the kit instructions.

[0330] ​​​Collecting culture 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.

[0331] The secretion of mouse TNF-α was detected by enzyme-linked immunosorbent assay (ELISA) kit. According to the operation steps of the kit, the specific steps are as follows:

[0332] (1) Reagent preparation

[0333] ①. Take out from the refrigerator and place at room temperature for 20 minutes. ②. Dilute the wash solution (20X) with double distilled water to 1X to prepare the required wash solution. ③. Add the standard diluent to the standard solution according to the volume marked on the label, and incubate at room temperature for 15 minutes. ④. Take 5 clean 1.5 mL centrifuge tubes, and add 250 μL of standard diluent to each tube for standard dilution. Finally, six standard concentrations are obtained, and the diluted standard is added to the pre-coated plate wells in turn. The standard diluent is directly added as 0 pg / mL concentration, and there are seven standard concentrations.

[0334] (2) Operation steps

[0335] ①. Calculate the number of pre-coated plates required for one experiment, and take the required plates and place them in a 96-well frame.

[0336] ②. Add 100 μL of sample or different concentrations of standard to the corresponding wells, cover the reaction wells with sealing film (transparent), and incubate at room temperature for 120 minutes.

[0337] ③. Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0338] ④. Add biotinylated antibody 100 μL / well, cover the reaction wells with sealing film (transparent), and incubate at room temperature for 60 minutes.

[0339] ⑤. Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0340] ⑥. Add 100 μL of horseradish peroxide labeled Streptavidin to each well, cover the reaction wells with sealing film (white), and incubate at room temperature in the dark for 20 minutes.

[0341] ⑦. Wash the plate 5 times, and the last time is placed on thick absorbent paper to dry.

[0342] ⑧. Add 100 μl of color developing agent TMB solution to each well, cover the reaction wells with sealing film (white), and incubate at room temperature in the dark for 20 minutes.

[0343] ⑨. Add 50 μL of stop solution to each well, mix well, and immediately measure the A450 value.

[0344] The test results are shown in Table 4. According to the test results of the mouse monocyte macrophage RAW 264.7 and the enzyme activity, the compounds A5, D2 and A4 can effectively inhibit the expression level of TNF-α. The above experimental results show that the compounds A5, D2 and A4 of the present application have good anti-inflammatory activity and can be used for the research of anti-inflammatory drugs.

[0345] Table 4 Anti-inflammatory activity results of the compounds of the present application on RAW 264.7 cells

[0346] Activity Example 4

[0347] The compounds A4, A5, D1 and D2 of the present application with better anti-inflammatory activity and enzyme activity were detected for cytotoxicity and inflammatory factor IC 50 .

[0348] 1. Experimental materials

[0349] (1) Experimental cells

[0350] Human immortalized epidermal cells HaCaT and mouse monocyte macrophage Raw 264.7

[0351] (2) Cell culture

[0352] The cells were cultured in DMEM (Gibco, Thermo Fisher Scientific) with the addition of 10% fetal bovine serum (Fetal Bovine Serum, Gibco, Thermo Fisher Scientific) and cultured at 37°C in 5% carbon dioxide. The culture medium was replaced every 2 days, and subculture was performed when the cells reached 90% confluence.

[0353] (3) Experimental instruments

[0354] Heating stirrer, paraffin sectioning machine, microscope, camera, syringe, enzyme label instrument, balance, oven, refrigerated centrifuge, etc.

[0355] Table 5

[0356] As shown in Table 5, the compounds A4, A5, D1 and D2 significantly down-regulated the expression of TNF-α in Raw264.7 and HaCaT cells. It is shown that the compounds A4, A5, D1 and D2 have certain anti-inflammatory effect and no toxicity to cells.

[0357] Activity Example 5

[0358] In vitro liver microsomal metabolic stability experiments were performed on 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(2,5-difluorobenzyl)-1 H- imidazole-4-carboxamide (A5) and 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(pyrimidin-2-ylmethyl)thiazole-4-carboxamide (D2) of the present application.

[0359] The 0.5 μM compound was incubated with liver microsomes (1 mg / mL) at 37 °C. 100 μL of reaction solution was taken at each time point of 0, 5, 15, 30, 45 min and 60 min, respectively. 200 μL acetonitrile containing internal standard was added to 100 μL reaction solution to extract the compound to be tested. The resulting mixture was centrifuged and the supernatant was analyzed by LC-MS / MS. The results are shown in Table 6, which shows that D2 has a half-life of 296.1 min in human liver microsomes and a half-life of 28.6 min in mouse liver microsomes, and has good metabolic stability.

[0360] Table 6

[0361] Active Example 6

[0362] In vitro Caco-2 permeability experiments were performed on 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(2,5-difluorobenzyl)-1 H-imidazole-4- carboxamide (A5) and 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N- (pyrimidin-2-ylmethyl)thiazole-4-carboxamide (D2) of the present application.

[0363] Caco-2 monolayer assays were performed for compounds A5, D2, respectively, using a standard procedure previously reported. Transport of the compounds was measured simultaneously from apical to basolateral (A-B) and in the opposite direction (B-A) under identical conditions. Propranolol and nadolol were used as high and low permeability controls, respectively. Digoxin was used as a positive control for PgP-mediated drug efflux. After washing the monolayers three times with Hanks Balanced Salt Solution (HBSS, Sigma-Aldrich), compound A5 or D2 was added to the appropriate wells (apical pH 6.8, basolateral pH 7.4). Incubation was performed at 37°C for 95 min. Samples were collected from the donor side at 5 min and 95 min, and from the acceptor side at 35 min and 95 min after incubation. Concentrations of samples were determined using liquid chromatography-mass spectrometry (LC-MS) / mass spectrometry. Mean values are the average of three independent experiments, each performed in triplicate.

[0364] Table 7

[0365] The results show that compound D2 has moderate cell permeability for the cells.

[0366] Note: a: Papp values in A to B (A-B) or B to A (B-A) were calculated by the following equation: Papp = (VA / (area x time)) x (drug acceptor / drug initial donor), where VA = volume in the acceptor well (in this assay: apical 0.1, basolateral 0.3 mL), area = surface area of the membrane (in this experiment: 0.143 cm 2 ), time = total transport time in seconds (in this experiment: 7200 s), and Papp values are expressed as 10 -6 cm / s. b: B-A / A-B ratio was calculated by dividing the Papp value from B to A by the Papp from A to B, as follows. Efflux ratio (B-A / A-B) = Papp (B to A) / Papp (A to B).

[0367] Active Example 7

[0368] In vivo pharmacokinetic experiments were performed for the compounds 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(2,5-difluorobenzyl)-lH-imidazole-4- carboxamide (A5) and 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N- (pyrimidin-2-ylmethyl)thiazole-4-carboxamide (D2) of the present application.

[0369] Pharmacokinetic experiments were performed for each compound with 6 five- to six-week-old male ICR mice, each weighing 18-22 g. Specifically, 6 ICR mice were randomly divided into 2 groups. Compound A5 or D2 was first dissolved in phosphate buffered saline containing DMSO and 5% Tween-80, and then administered orally (PO, 10 mg / kg) and intravenously (IV, 5 mg / kg) after 12 h of fasting, with feeding after 4 h of administration. Blood was collected at each time point, with IV blood collection points (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 24 h) and PO blood collection points (15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h). Blood was collected from the submandibular vein or other suitable means, with each sample being collected at about 30 μL / time point, and K2-EDTA was used for anticoagulation. After blood sample collection, the blood was placed on ice, and the plasma was separated by centrifugation (centrifugation conditions: 6800 g, 6 min, 2-8 °C) within 1 h. The plasma samples were stored in a -80 °C refrigerator before analysis. LC-MS / MS (Agilent 1260 Infinity LC system and Agilent 6460 triple quadrupole mass spectrometer system) was used to analyze the plasma concentration of the ICR mice.

[0370] Table 8 Pharmacokinetic data of compound A5 in ICR mice

[0371] Table 9 Pharmacokinetic data of compound D2 in ICR mice

[0372] Active Example 8

[0373] The in vivo efficacy evaluation of the mouse anti-psoriasis of 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(2,5-difluorobenzyl)-1H-imidazole-4-carboxamide (A5) and 2-(3-(cyclopropylmethoxy)-4-(difluoromethoxy)phenyl)-N-(pyrimidin-2-ylmethyl)thiazole-4-carboxamide (D2) of the present application was carried out.

[0374] I. Experimental purpose

[0375] To investigate the therapeutic effect of compounds A5 and D2 on imiquimod-induced mouse psoriasis.

[0376] II. Experimental materials

[0377] Experimental animals: Balb / c male mice, 56 for each compound, age: 6-8 weeks; weight: about 18-22g.

[0378] Animal grouping, modeling and sampling

[0379] Experimental instruments: heating stirrer, paraffin sectioning machine, microscope, syringe, enzyme label instrument, balance, oven, refrigerated centrifuge, etc.

[0380] Experimental reagents: HE staining related reagents, PBS solution, etc., imiquimod, ELISA kit of IL-17A, IL-1β and IL-6.

[0381] III. Animal experiment

[0382] 1. Establishment of mouse psoriasis model and drug administration

[0383] ① The back of the mouse was shaved in an area of 2cm×3cm three days before modeling. The shaving effect was observed, and the fur and new hair that grew out were shaved off at the beginning of modeling.

[0384] ② The mice were randomly divided into 7 groups: blank group, model group (5% IMQ 62.5mg / d, continuously for 9 days), compound group (0.3%, 0.6%, 1.2%), positive control group (Roflumilast, 0.3%), auxiliary material group (only cream auxiliary material formula was applied), 8 in each group, a total of 56.

[0385] Blank group: from the first day of psoriasis modeling, no treatment was given to the mice.

[0386] Model group: from the first day, 62.5 mg of 5% IMQ was applied to the back of the mice to establish a psoriasis model, and the application was continued for 9 days.

[0387] Positive treatment control group: from the first day of psoriasis modeling, 62.5 mg of 5% IMQ was applied to the back of the mice, and from the third day, 62.5 mg of 5% IMQ was applied to the back of the mice in the morning, and 4 hours later, 62.5 mg of roflumilast cream (0.3%) was applied to each mouse, and the application was continued for 7 days.

[0388] Compound group (0.3%, 0.6%, 1.2%): from the first day of psoriasis modeling, 62.5 mg of 5% IMQ was applied to the back of the mice, and from the third day, 62.5 mg of 5% IMQ was applied to the back of the mice in the morning, and 4 hours later, low, medium, and high doses of compound cream were applied to each group of mice for treatment, and the application was continued for 7 days.

[0389] Adjuvant group: from the first day of psoriasis modeling, 62.5 mg of adjuvant cream was applied to the back of the mice, and the application was continued for 7 days.

[0390] 2. Experimental operation and results

[0391] 2.1 Scoring of the mouse psoriasis area and severity index (PSAI)

[0392] The degree of scaling, the size of erythema, and the degree of skin thickening on the back of the mice were observed by naked eye, and the PASI score was processed. The PASI score rule: the severity of psoriasis was evaluated from three angles of erythema, scaling, and thickening. After scoring, the skin of the mice was photographed.

[0393] As shown in FIG. 1, the model group mice showed a typical psoriasis-like dermatitis phenotype, with redness, scales, and thickness. Application of compound A5 significantly reduced the symptoms of psoriasis.

[0394] As shown in FIG. 2, the model group mice showed a typical psoriasis-like dermatitis phenotype, with redness, scales, and thickness. Application of compound D2 significantly reduced the symptoms of psoriasis.

[0395] As shown in FIG. 3, compound A5 (1.2%) had the same efficacy as roflumilast in terms of redness score, thickness, and total score. No significant weight loss was observed, indicating that A5 has a low risk of toxicity in vivo.

[0396] As can be seen from Figure 4, the efficacy of compound D2 (0.6%, 1.2%) in terms of scores of erythema score, thickness, total score, and body weight has the same efficacy as that of roflumilast, and the risk of toxicity in vivo is low.

[0397] 2.2 Mouse sacrifice

[0398] Euthanasia was performed by intraperitoneal injection of an overdose of 0.3% sodium pentobarbital solution.

[0399] 2.3 Paraffin embedding and sectioning of skin tissue

[0400] 1) Cut the full-thickness skin parallel to the vertebral line and place it in 10% neutral formaldehyde solution for fixation for more than 24 h;

[0401] 2) Place the trimmed tissue block in an embedding box and wash it with running water for 24 h to completely remove residual formaldehyde;

[0402] 3) Dehydrate the tissue by alcohol gradient, the specific steps are as follows: 70% alcohol overnight, 80% alcohol for 1.5 h, 95% alcohol I for 45 min, 95% alcohol II for 30 min, 100% alcohol I for 25 min, and 100% alcohol II for 20 min;

[0403] 4) After dehydration, place the tissue in an alcohol / dichloromethane (1:1, v / v) solution for 20 min;

[0404] 5) Dichloromethane transparency I and II for 20 min and 10 min;

[0405] 6) Place the tissue in preheated melted paraffin I, II and embedding paraffin in a 60-65°C oven for 1 h each;

[0406] 7) Pour a small amount of embedding paraffin into a preheated metal embedding frame, place the skin tissue block in it vertically to the embedding frame, pour paraffin again, embed the tissue, and cool it down;

[0407] 8) Paraffin sectioning: Fix the tissue paraffin block on a Leica microtome. The section thickness is 5 μM, and continuous sectioning is performed. Place the section in 40°C water using a toothless forceps for expansion, and then use a glass slide to fish the section. After baking the section at 60°C for 2 h, store it in a section box at room temperature for standby.

[0408] 2.4 HE staining

[0409] 1) De-waxing and hydration: Place the skin tissue paraffin section in dichloromethane I and II for 15 min each, and then in 100% ethanol I and II for 3 min each, 95% ethanol I and II for 3 min each, 80% ethanol for 3 min, and double distilled water for 1 min;

[0410] 2) Hematoxylin staining for 15 min, washing with water, and loading the slide with excess dye;

[0411] 3) 1% hydrochloric acid ethanol (99 mL 70% ethanol + 1 mL concentrated hydrochloric acid) for 3 s, and the nucleus and chromatin were observed under a microscope to be clear;

[0412] 4) Rinse with running water for 15 min, and distilled water for 1 min;

[0413] 5) Eosin for 2 min, and running water for 1 min;

[0414] 6) Dehydration with 80% and 100% ethanol for 2 s and 7 min, respectively;

[0415] 7) Xylene I and II for 5 min each;

[0416] 8) Mounting: the slide was taken out of xylene II, neutral balsam was added at the tissue, a cover glass was gently placed on top, and natural air drying was performed.

[0417] 9) Observation: pathological changes were observed under a microscope, and photographs and analysis were performed.

[0418] As shown in FIGS. 5 and 6, the skin thickness of the model group was thickened, and a large number of inflammatory cells infiltrated, and the dosed groups (A5, D2) had a dose-dependent protective effect on the skin.

[0419] 2.5 Eight mice in each group were taken for the spleen, and A5 and D2 were photographed and recorded.

[0420] The spleen is the largest immune organ in the human body, contains a large number of immune cells, and the body produces an inflammatory response to a certain extent and has a great connection with the immune system. In the experiment, the spleen of each treatment group was observed, and the weight of the spleen was measured. The spleen of the model group was enlarged, and the spleen of the dosed group was significantly reduced, and it was found that the dosed group (A5, D2) had an inhibitory effect on the inflammatory infiltration of the spleen, see FIGS. 7 and 8.

[0421] Note: The weight index of the mouse spleen: Spleen index = Spleen weight (mg) / Body weight (g) on the last day.

[0422] 2.6 Ki-67 antibody immunohistochemical staining

[0423] 1) The skin tissue paraffin section was placed in an oven, and baked at 60°C for 1 h.

[0424] 2) Dewaxing: xylene for 10 min three times → anhydrous ethanol I for 5 min → anhydrous ethanol II for 5 min → 95% ethanol for 5 min → 85% ethanol for 5 min → 75% ethanol for 5 min → ddH2O for 5 min.

[0425] 3) Antigen retrieval: 400 ml of antigen retrieval solution (800 ml of dd H2O + 3 g of sodium citrate + 400 mg of citric acid, then make up to 1000 ml) is boiled in advance, the slices are placed in a beaker and boiled for 20 min, after being placed at room temperature, the slices are washed with PBS for 3 min each time.

[0426] 4) Remove PBS, take out the slices from PBS, spin the slices twice, mark with an immunohistochemical pen, then place them in PBS, spin the slices again after marking, add 3% hydrogen peroxide dropwise, incubate at room temperature for 5 min, wash the slices with PBS for 3 min each time.

[0427] 5) Remove PBS, add 10% goat serum blocking solution dropwise, and incubate at room temperature for 1 h.

[0428] 6) Remove the goat serum, spin dry, and then directly add the primary antibody dropwise, and incubate the wet box at 4°C (about 14 h).

[0429] 7) Take out the slices, rewarm for 30 min, and wash with PBS for 3 min each time.

[0430] 8) Remove PBS, add the secondary antibody for immunohistochemistry to each sample dropwise, incubate at room temperature for 15 min, and wash with PBS for 3 min each time.

[0431] 9) Spin the slices dry, immediately place them under a microscope, add DAB staining dropwise, and wash with PBS for 5 min after color development.

[0432] 10) Hematoxylin counterstaining for 40 s → rinse with running water for 2 min → return to blue for 7 s → rinse with running water for 10-15 min → soak in dd H2O for 5 min.

[0433] 11) 95% ethanol (1 min) → anhydrous ethanol I (1 min) → anhydrous ethanol II (1 min) → xylene (1 min) → xylene (1 min) → xylene (1 min) neutral resin mounting.

[0434] The results show (see FIGS. 9 and 10) that the expression of Ki-67 antibody in the model group is significantly increased, while the expression of Ki-67 antibody in the drug groups (A5, D2) is significantly reduced, indicating that the compounds have a significant inhibitory effect on skin thickening.

[0435] Four, expression of inflammatory factors in the skin of psoriasis mice after different treatments.

[0436] ① Protein levels of IL-17A, TNF-a, IL-6, and IL-1b in skin lesions

[0437] 1) Take 100 mg of the lesioned skin into an ep tube, add 1 mL of normal saline, and grind using a tissue grinder.

[0438] 2) 4℃, 3500rpm and 12000rpm each centrifugation once, take supernatant, use BCA method to determine the protein concentration in tissue grinding liquid, ELISA kit to determine the content of IL-1β, IL-6, IL-17A and TNF-α, according to the kit instructions.

[0439] The results are shown in Figure 11, showing that compound A5 has a down-regulation effect on IL-1β, IL-6, IL-17A and TNF-α inflammatory factors, indicating that compound A5 has the effect of treating psoriasis-related inflammation.

[0440] The results of Figure 12 show that compound D2 has a down-regulation effect on IL-1β, IL-6, IL-17A and TNF-α inflammatory factors, indicating that compound D2 has the effect of treating psoriasis-related inflammation.

Claims

1. A compound of Formula I or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof wherein R1and R2are each independently H, and R1and R2are not simultaneously H; X and Y are each independently N or NH, O, or S, at least one of X and Y being N; Z is NH or O; R3is C6-14aryl, 5-14 membered heteroaryl, C6-10cycloalkyl, or 5-14 membered heterocyclyl; optionally, the C6-14aryl, 5-14 membered heteroaryl, C6-10cycloalkyl, or 5-14 membered heterocyclyl is substituted with one or more substituents selected from halogen, halogenated C1-C6alkyl, carbonyl, C1-C6alkyl, and C1-C6alkoxy; the 5-14 membered heteroaryl or 5-14 membered heterocyclyl comprises 1-3 heteroatoms selected from N, O, and S; preferably, the halogen is F, Cl, Br, or I; n is 0, 1, 2, or 3; each dotted line independently represents a bond that is present or absent.

2. The compound of claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof, wherein R1 is R2 is H; Preferably, R1 is R2is H; Preferably, R1 is R2 is Preferably, R1 is R2 is 3. The compound of claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof, wherein X is N, Y is NH; Preferably, X is N, Y is O; Preferably, X is N, Y is S.

4. The compound of claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof, wherein R3is C6-10aryl, 5-10 membered heteroaryl, C6-10cycloalkyl, or 5-10 membered heterocyclyl; optionally, the C6-10aryl, 5-10 membered heteroaryl, C6-10cycloalkyl, or 5-10 membered heterocyclyl is substituted with one or more substituents selected from F, carbonyl, C1-C4alkyl, and C1-C4alkoxy; the 5-10 membered heteroaryl or 5-10 membered heterocyclyl comprises 1-3 heteroatoms selected from N, O, and S; Preferably, R3 is 5. The compound of claim 1 or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof, wherein Formula I is R4is n is 0 or 1 ; Preferably, formula I is wherein each R5is independently n is 0 or 1; Preferably, formula 1 is R6 is n is 0 or 1; Preferably, formula I is R7 is n is 0 or 1; Preferably, formula I is R8 is n is 0 or 1; Preferably, formula I is R9 is n is 0 or 1.

6. A compound or a pharmaceutically acceptable salt, solvate, hydrate, prodrug, stereoisomer, polymorph, co-crystal, or deuterated form thereof:

7. An intermediate compound which is the following compound:

8. A process for the preparation of a compound of formula II, ###00010### II wherein R1, R2, X, Y, R3, n are as described in any one of claims 1-5; the preparation method comprising: (1) wherein a) reacting 3,3-dibromo-1,1,1-trifluoro-2-ketone with sodium acetate at 80-120 °C for 1-2 hours, adding a solution of benzaldehyde substituted with R1, R2, difluoromethoxy, reacting at 20-50 °C for 1-8 hours; b) hydrolyzing the product obtained in step a) at 50-90 °C for 1-8 hours; c) reacting the product from step b) with R3-(CH2) n -NH2 at 20 °C to 50 °C for 5 to 7 hours; (2) wherein d) reacting the benzoic acid substituted with R1, R2, difluoromethoxy with L- serine methyl ester hydrochloride and SOCl2 at 0 °C for 10-14 hours; e) stirring the product obtained in step d) with DAST at -78 °C for 3-5 hours, adding an inorganic base, reacting at 20-50 °C for 24 hours; f) reacting the product obtained in step e) with CBrCl3 and DBU at 0 °C for 15-24 hours; g) hydrolyzing the product obtained in step f) at 20-50 °C for 1-8 hours; h) reacting the product from step g) with R3-(CH2) n -NH2 at 20 °C to 50 °C for 3 to 8 hours; or (3) i) stirring benzaldehyde substituted by R1, R2, difluoromethoxy with D-cysteine methyl ester hydrochloride, K2CO3 at 20-50 °C for 20-30 hours, then stirring at -15 °C, then adding DBU and CBrCl3 at -20-0 °C, reacting at 20-50 °C for 12-24 hours; j) hydrolyzing the product obtained in step i) by stirring at 20-50 °C for 8-12 hours; k) reacting the product from step j) with R3-(CH2) n -NH2 at 20 °C to 50 °C for 4 to 8 hours.

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

10. Use of a compound of any one of claims 1-6 or a pharmaceutical composition of claim 9 in the manufacture of a medicament for preventing and / or treating an inflammatory disease, a respiratory disease, a skin disease, or an immune system disease; more preferably, the inflammatory disease is an inflammatory skin 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 a compound of any one of claims 1-6 or a pharmaceutical composition of claim 9 in the manufacture of a medicament for preventing and / or treating a PDE4-mediated disease or a PDE4 inhibitor.

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