Aromatic amide compound, pharmaceutical composition and use thereof

Abstract

The present invention discloses an aromatic amide compound, a pharmaceutical composition and its use, and belongs to the field of chemical medicine. The present invention provides an aromatic amide compound with a structure as shown in general formula (I) and a pharmaceutically acceptable salt thereof, which can be used as a Bruton's tyrosine kinase (BTK) inhibitor, and can be used to prevent and / or treat cancer based on BTK inhibition.

Description

Technical Field

[0001] The present invention belongs to the field of chemical medicine, and relates to an aromatic amide compound, a pharmaceutical composition and use thereof. Background Art

[0002] Bruton's tyrosine kinase (BTK) is an important enzyme that mediates cell signal transduction and is present in plasma cells, including B cells. B cells are activated through the B cell receptor (BCR), and BTK plays a decisive role in the BCR-mediated signaling pathway. After the BCR on the B cell is activated, it causes the activation of BTK, leading to an increase in the concentration of downstream phospholipase C (PLC) and activation of the IP3 and DAG signaling pathways. This signaling pathway can promote cell proliferation, adhesion and survival, and plays an important role in the development of B cell lymphoma.

[0003] The presence of BTK mutants in the population proves that BTK can be a drug target. X-linked agammaglobulinemia (XLA) is a rare genetic disease that exists in 1 / 250,000 men. In this disease, the function of BTK is inhibited, resulting in the blockage of B cell production or maturation. Men with XLA disease have basically no B cells in their bodies, few circulating antibodies, and are prone to serious or even fatal infections. This shows that BTK plays an extremely important role in the growth and differentiation of B cells.

[0004] BTK inhibitors can inhibit the proliferation of B lymphoma cells, destroy the adhesion of tumor cells, and promote the apoptosis of tumor cells by inhibiting the activity of BTK, making BTK an attractive drug target in B cell-related cancers, especially for B cell lymphomas and leukemias, such as non-Hodgkin's lymphoma, chronic lymphocytic leukemia, and relapsed or refractory mantle cell lymphoma.

[0005] Ibrutinib, an approved irreversible BTK inhibitor, also irreversibly binds to interleukin-2 inducible tyrosine kinase (ITK). ITK plays a key role in FcR-stimulated natural killer (NK) cell function required for antibody-dependent NK cell-mediated cytotoxicity (ADCC). ADCC is a mechanism that anti-CD20 antibodies such as rituximab are thought to activate, and ibrutinib has been shown to antagonize this mechanism in vitro. Since rituximab combined with chemotherapy is the standard for the treatment of B-cell malignancies today, it is hoped that there will be a BTK inhibitor that is more selective for BTK than ITK. In the clinic, adverse events include atrial fibrillation, diarrhea, rash, arthralgia, and bleeding. Known BTK inhibitors such as ibrutinib are also thought to have gastrointestinal side effects, which are thought to be the result of secondary EGFR inhibitory activity. Therefore, a BTK inhibitor with high BTK inhibition and low EGFR inhibition is needed to reduce or avoid gastrointestinal side effects.

[0006] In the clinic, adverse events include atrial fibrillation, diarrhea, rash, arthralgia and bleeding. Known BTK inhibitors such as ibrutinib are also considered to have gastrointestinal side effects, which are considered to be the result of secondary EGFR inhibitory activity. Therefore, a BTK inhibitor with high BTK inhibition and low EGFR inhibition is needed to reduce or avoid gastrointestinal side effects. Summary of the invention

[0007] The inventors have discovered a new substituted pyrazole compound in the process of studying Bruton's tyrosine kinase (BTK) inhibitors, which has excellent inhibitory activity on BTK and lower toxicity and side effects, and provides a treatment plan for patients with BTK inhibitors that are resistant to C481 mutations, with a longer half-life and better safety. It is expected that such inhibitors will have good therapeutic effects, are expected to overcome drug resistance and toxicity problems, and have good development prospects.

[0008] The present invention provides an aromatic amide compound or a pharmaceutically acceptable salt thereof, a pharmaceutical composition and use thereof. The compound can be used as a BTK inhibitor and has better pharmacodynamics and higher metabolic stability.

[0009] In one aspect, the present invention provides an aromatic amide compound having a structure as shown in general formula (I) or a pharmaceutically acceptable salt thereof:

[0010]

[0011] Where:

[0012] Ring A is R1 and R2 are independently selected from hydrogen or deuterium; R3 is selected from C1-C4 straight chain or branched alkyl, C3-C6 cycloalkyl or methyl substituted with 1-3 deuterium atoms; R4 is selected from H, C1-C4 alkyl substituted with 0-3 deuterium atoms, C3-C6 cycloalkyl substituted with 0-3 deuterium atoms, C1-C4 alkyl substituted with halogen atoms;

[0013] Alternatively, ring A is R3 is selected from C1-C4 straight or branched alkyl, C3-C6 cycloalkyl or methyl substituted with 1-3 deuterium atoms; R1, R2, R5 are independently selected from hydrogen or deuterium, and at least one is deuterium.

[0014] In certain embodiments of the present invention, R3 is preferably selected from methyl or deuterated methyl (-CD3).

[0015] In certain embodiments of the present invention, R4 can be specifically selected from H, methyl, methyl substituted with 1-3 deuterium atoms, CF3. In certain embodiments of the present invention, alkyl refers to a straight chain or branched alkyl. Halogen atoms include F, Cl, Br. In certain embodiments of the present invention, the compound is selected from but not limited to:

[0016]

[0017] In certain embodiments of the present invention, the pharmaceutically acceptable salt is an inorganic salt or an organic salt. The inorganic salt includes hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from formates, acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, succinates, glutarates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, p-toluenesulfonates, and ascorbates.

[0018] The present invention further relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture thereof, and pharmaceutically acceptable salts thereof.

[0019] In one embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0020] In one embodiment of the present invention, the pharmaceutically acceptable carrier includes: microspheres, nanoparticles and liposomes.

[0021] In one embodiment of the present invention, the pharmaceutical composition further comprises pharmaceutical excipients.

[0022] In one embodiment of the present invention, the pharmaceutical excipients include: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, flocculants and deflocculating agents, filter aids and release retardants.

[0023] In one embodiment of the present invention, the dosage forms of the pharmaceutical composition include: injection, lyophilized powder for injection, suspension, implant, embolic agent, capsule, tablet, pill and oral solution.

[0024] on the other hand,

[0025] The present invention also provides the use of the above aromatic amide compounds or pharmaceutically acceptable salts thereof in the preparation of BTK kinase agents.

[0026] The present invention also provides use of the aromatic amide compounds or pharmaceutically acceptable salts thereof in preparing a drug for treating a disease regulated by Bruton's tyrosine kinase (BTK).

[0027] The compounds of the invention are useful for treating conditions modulated by Bruton's tyrosine kinase (BTK). Conditions modulated by BTK are generally conditions that can be treated by inhibiting BTK using the compounds of the invention. Compounds of any formula of the invention are useful for treating conditions that can be treated by inhibiting Bruton's tyrosine kinase (BTK).

[0028] The conditions modulated by BTK described herein include cancer, lymphoma, leukemia, autoimmune disease, inflammatory disorder, heteroimmune disease or fibrosis.

[0029] The present invention also provides a method for treating BTK-related diseases or diseases associated with undesirable BTK activity, particularly allergic diseases, autoimmune diseases (e.g., rheumatoid arthritis), inflammatory diseases or cancers (e.g., B cell proliferative disorders, such as chronic lymphocytic lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, follicular lymphoma or chronic lymphocytic leukemia), which generally comprises administering an effective amount of a compound of the present invention, its N-oxide or a prodrug thereof to a mammal in need, and optionally detecting the improvement of the disease or symptom, or BTK inhibition. Another aspect of the present invention also provides a method for preparing a compound of formula (I), which is implemented by one of the following reaction schemes:

[0030] The compounds of the present invention and their pharmaceutically acceptable salts can be prepared from the following materials: (a) commercially available starting materials (b) known starting materials that can be prepared as described in the literature process (c) novel intermediates described in the schemes and experimental procedures herein. In the process of preparing the compounds of the present invention, the order of the synthetic steps can be changed to increase the yield of the desired product. Some compounds in the present invention can be produced by the following reaction schemes and their descriptions.

[0031] Reaction scheme 1:

[0032]

[0033] As shown in reaction formula 1, p-bromobenzoyl chloride S1 is used as a raw material, and reacts with malononitrile S2 under the action of a base to obtain an intermediate S3; intermediate S3 undergoes a substitution reaction with dimethyl sulfate or methyl iodide to obtain intermediate S4; intermediate S4 and the corresponding hydrazine derivative S5 are cyclized to obtain intermediate S6; intermediate S6 and the corresponding substituted potassium borate intermediate S7 are coupled to obtain intermediate S8; intermediate S8 is hydrolyzed under acidic conditions to obtain the corresponding compound S9; wherein R1 and R2 are both hydrogen or both deuterium; R3 is selected from a linear or branched C1-C4 alkyl, C3-C6 cycloalkyl or a methyl substituted with 1-3 deuterium atoms; R4 is selected from H, a linear or branched C1-C4 alkyl, C3-C6 cycloalkyl or a methyl substituted with 1-3 deuterium atoms;

[0034] or

[0035] Reaction scheme 2:

[0036]

[0037] Reaction 2

[0038] As shown in reaction formula 2, intermediate S6 is used as a raw material and Boc protection is performed to obtain intermediate S10; intermediate S10 is catalyzed by metal in a CO atmosphere to obtain intermediate S11; intermediate S11 reacts with the corresponding intermediate S12 to form a Schiff base, and then intermediate S13 is obtained under the action of sodium borodeuteride, and intermediate S13 is hydrolyzed by cyanide under acidic conditions to obtain the corresponding compound S14; wherein, R3 is selected from a linear or branched C1-C4 alkyl, a C3-C6 cycloalkyl or a methyl substituted with 1-3 deuterium atoms, and R4 is selected from H, a linear or branched C1-C4 alkyl, a C3-C6 cycloalkyl or a methyl substituted with 1-3 deuterium atoms;

[0039] or

[0040] Reaction scheme three:

[0041]

[0042] As shown in reaction formula 3, intermediate S15 and intermediate S4 undergo ring-closure reaction to obtain intermediate S16, the ester of intermediate S16 is reduced in a corresponding reducing agent to obtain intermediate S17, intermediate S17 is oxidized to obtain intermediate S18, intermediate S18 undergoes intramolecular ring-closure by reductive amination to obtain intermediate S19, intermediate S19 is coupled with the corresponding substituted intermediate S7 to obtain intermediate S20; intermediate S20 undergoes cyanohydrolysis under acidic conditions to obtain the corresponding compound S21; wherein R1 and R2 are both hydrogen or both deuterium; R3 is selected from a linear or branched C1-C4 alkyl group, a C3-C6 cycloalkyl group or a methyl group substituted with 1-3 deuterium atoms; R4 is selected from H, a C1-C4 alkyl group, a C3-C6 cycloalkyl group, a methyl group substituted with 1-3 deuterium atoms or CF3;

[0043]

[0044] As shown in reaction formula 4, intermediate S22 is used as a raw material and Boc protection is performed to obtain intermediate S23; intermediate S23 is catalyzed by metal in a CO atmosphere to obtain intermediate S24; intermediate S24 reacts with the corresponding S12 to form a Schiff base, and then intermediate S25 is obtained under the action of sodium borodeuteride; intermediate S25 is hydrolyzed by cyano group under acidic conditions to obtain the corresponding compound S26; wherein R3 is selected from C1-C4 alkyl, C3-C6 cycloalkyl or methyl substituted with 1-3 deuterium atoms, and R4 is selected from H, C1-C4 alkyl, C3-C6 cycloalkyl, methyl substituted with 1-3 deuterium atoms or CF3;

[0045] The specific reaction conditions of the above reaction can refer to the conditions in the following examples.

[0046] Beneficial effects:

[0047] The present invention provides a compound for inhibiting Bruton's tyrosine kinase (BTK). In addition, the inhibitor of the present invention has better pharmacodynamic performance and higher metabolic stability, and can effectively reduce side effects such as rash and diarrhea. DETAILED DESCRIPTION

[0048] The technical solution of the present invention will be described in detail below in conjunction with embodiments.

[0049] As used herein, the term "disease" refers to any condition or disorder that damages or interferes with the normal function of a cell, organ, or tissue.

[0050] As used herein, the term "inhibitor" refers to a compound or agent that has the ability to inhibit the biological function of a targeted protein or polypeptide, for example, by inhibiting the activity or expression of the protein or polypeptide.

[0051] As used herein, the term "anti-neoplastic agent" refers to any agent useful in the treatment of neoplastic disorders.

[0052] As used herein, the term "pharmaceutically acceptable" refers to a component that is suitable for contact with the tissues of humans and other mammals without excessive toxicity, irritation, allergic response, etc., within a reasonable medical range and has a reasonable benefit / risk ratio. "Pharmaceutically acceptable salt" refers to any non-toxic salt that, after administration to a recipient, can directly or indirectly provide a compound of the present invention or a prodrug of the compound.

[0053] As used herein, the term "effective amount" or "effective therapeutic amount" refers to an amount of a compound or pharmaceutical composition described herein that is sufficient to achieve the intended application, including, but not limited to, treating a disease. In some embodiments, the amount is detected to be effective for killing or inhibiting cancer cell growth or spread; the size or number of tumors; or the severity level, stage and progression of cancer. The effective therapeutic amount may vary depending on the intended application, such as in vitro or in vivo, the condition and severity of the disease, the age, weight, or mode of administration of the subject, etc. The term also applies to doses that will induce a specific response in target cells, such as reducing cell migration. The specific dose will depend on, for example, the specific compound selected, the subject species and their age / existing health condition or health condition risk, the route of administration, the severity of the disease, administration in combination with other agents, the time of administration, the tissue to which it is administered, and the drug delivery device, etc.

[0054] "Administration" or "administering" a subject compound in the context of the present invention means providing the subject in need of treatment with a compound of the present invention.

[0055] The compounds of the present invention may contain one or more asymmetric centers and therefore occur as racemates and racemic mixtures, single enantiomers, individual diastereomers and diastereomeric mixtures. All such isomeric forms of these compounds are expressly included in the present invention. The compounds of the present invention may also exhibit multiple tautomeric forms, in which case, the present invention expressly includes all tautomeric forms of the compounds described herein. All such isomeric forms of such compounds are included in the present invention. All crystalline forms of the compounds described herein are expressly included in the present invention.

[0056] The following examples are used to illustrate but not to limit the synthesis of compounds of formula (I). All temperatures are in degrees Celsius. Unless otherwise stated, all evaporations were carried out under reduced pressure. Unless otherwise stated, reagents were purchased from commercial suppliers and used without further purification. The structures of the final products, intermediates and raw materials were confirmed by standard analytical methods, such as elemental analysis, spectral feature analysis, such as MS, NMR. The abbreviations used are conventional abbreviations in the art, and some intermediates were purchased from Yancheng Zhengchi Biotechnology Co., Ltd.

[0057] The following intermediate substances are involved in the specific embodiments, and can be synthesized by referring to the following route process:

[0058] 5-Amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide (C1)

[0059]

[0060] Step 1: Preparation of 2-[(4-bromophenyl)-hydroxy-methylene]malononitrile 1c

[0061] 4-Bromobenzoyl chloride 1a (100g, 456mmol) and malononitrile 1b (33.1g, 502mmol) were dissolved in toluene (1000mL) and THF (200mL) solution, nitrogen protection, cooled to -10℃, and N, N-diisopropylethylamine (117.6g, 912mmol) in toluene (500mL) solution was slowly added dropwise, keeping the internal temperature at about -10℃. After the addition was completed, the reaction mixture was stirred at 0℃ for 1 hour, and then stirred at room temperature for 18 hours. After the reaction was completed, hydrochloric acid (1M) and ethyl acetate were added, the layers were separated, and the aqueous layer was extracted with ethyl acetate. The combined organic layer was washed with HCl (1M) and saturated sodium chloride in turn, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2-[(4-bromophenyl)-hydroxy-methylene] malononitrile 1c (100g, yield: 87%) as a light yellow solid. MS-ESI(m / z):249,251[M+1] +

[0062] Step 2: Preparation of 2-[(4-bromophenyl)-methoxy-methylene]malononitrile 1d

[0063] A solution of 2-[(4-bromophenyl)-hydroxy-methylene]malononitrile 1c (100 g, 401 mmol) in THF (200 mL) was added dropwise to a suspension of sodium hydride (19.2 g, 482 mmol) in THF (800 mL) and cooled to 0°C. After stirring at 0°C for 30 minutes, dimethyl sulfate (152 g, 1209 mmol) was added, heated to 80°C and stirred for 16 hours. After the reaction was completed, it was cooled to room temperature, quenched with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was further purified by silica gel flash column chromatography to give 2-[(4-bromophenyl)-methoxy-methylene]malononitrile 1d (65 g, yield: 61%) as a white solid. MS-ESI (m / z): 263 [M+1] +

[0064] Step 3: Preparation of 5-amino-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 1f

[0065] 2-[(4-bromophenyl)-methoxy-methylene]malononitrile 1d (10 g, 38 mmol) and triethylamine (11.5 g, 114 mmol) were dissolved in EtOH (200 mL) solution, (1,1,1-trifluoropropane-2-yl) hydrazine hydrochloride (7.4 g, 45 mmol) was added, and then heated to 100 ° C and stirred for 8 h. After the reaction was completed, it was concentrated under reduced pressure. Saturated aqueous ammonium chloride solution and ethyl acetate were added to the residue, the organic layer was separated, and washed with saturated sodium chloride in turn, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave 5-amino-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 1f (8 g, yield: 58%) as a white solid. MS-ESI (m / z): 359 [M+1] +

[0066] Step 4: Preparation of N-(4-(5-amino-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide 1h

[0067] (1) Preparation of 1 g of potassium trifluoro-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]borane:

[0068]

[0069] Step A: Preparation of 5-fluoro-2-deuterated methoxybenzoic acid deuterated methyl ester 1g-2

[0070] 5-Fluorosalicylic acid 1g-1 (5g, 32mmol) and potassium carbonate (8.84g, 64mmol) were dissolved in DMF (50mL), deuterated iodomethane (11.6g, 80mmol) was added, and stirred at room temperature for 16h. After the reaction was completed, the reaction solution was diluted with ethyl acetate, washed with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was further purified by silica gel flash column chromatography to obtain deuterated methyl 5-fluoro-2-deuterated methoxybenzoate 1g-2 (5g, yield: 82%) as a colorless oil.

[0071] Step B: Preparation of 5-fluoro-2-deuterated methoxybenzoic acid 1g-3

[0072] Dissolve 5-fluoro-2-deuterated methoxybenzoic acid deuterated methyl ester 1g-2 (5g, 26.3mmol) in tetrahydrofuran (50mL) and water (50mL), add lithium hydroxide hydrate (3.3g, 78.9mmol), and then stir at room temperature for 16h. After the reaction is completed, concentrate under reduced pressure to remove tetrahydrofuran, and adjust the pH of the residual aqueous solution to 3-4 with 1M hydrochloric acid. Solid precipitates, filter, and dry to obtain 5-fluoro-2-deuterated methoxybenzoic acid 1g-3 (3.5g, yield: 77%) as a white solid. MS-ESI (m / z): 174 [M+1] +

[0073] Step C: Preparation of 5-fluoro-2-deuterated methoxybenzoyl chloride 1g-4

[0074] 5-Fluoro-2-deuterated methoxybenzoic acid 1g-3 (3.5 g, 20.2 mmol) was dissolved in thionyl chloride (20 mL), heated to 50°C for 2 hours. After the reaction was completed, it was concentrated under reduced pressure to obtain 5-fluoro-2-deuterated methoxybenzoyl chloride 1g-4 (3.6 g, yield: 93%) as a colorless oil.

[0075] Step D: Preparation of potassium trifluoro-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]borane 1 g

[0076] A 1M toluene solution of potassium bis(trimethylsilyl)amide (19.7 mL, 19.7 mmol) was added dropwise to a solution of 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (4.34 g, 19.7 mmol) in anhydrous THF (100 mL). After stirring at -78°C for 25 minutes, the mixture was stirred at 0°C for another 10 minutes and then at room temperature for 30 minutes. Anhydrous methanol (20 mL) was added at room temperature and a precipitate was formed. The mixture was stirred at room temperature for another 1 hour and then the reaction mixture was concentrated under reduced pressure. The residue was dissolved in anhydrous THF (50 mL) and then a solution of 5-fluoro-2-deuterated methoxy-benzoyl chloride 1g-4 (3.6 g, 18.8 mmol) in THF (25 mL) was slowly added. The reaction mixture was then stirred at room temperature for 16 hours and concentrated under reduced pressure after the reaction was completed. The obtained residue was dissolved in cold MeOH (50 mL), and the mixture was then cooled to 0 ° C, followed by addition of a saturated solution of potassium bifluoride (7.33 g, 94 mmol) in water (20 mL). The reaction mixture was naturally warmed to room temperature and stirred for 16 hours, then concentrated under reduced pressure. The residue was azeotroped with toluene (3X 50 mL) twice to remove water. The residue was then washed with cold TBME and filtered. The white solid was washed with cold acetone (50 mL) and cold TBME to obtain trifluoro-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]borane potassium 1g (3 g, yield: 54%), a white solid.

[0077] 1 H NMR (400MHz, DMSO-d6) δ: 7.80-7.71 (m, l H), 7.64 (dd, J=9.8, 3.4Hz, 1H), 7.31-7.25 (m, l H), 7.16 (dd, J=9.2, 4.4Hz, 1H), 2.17-2.09 (m, 2H).

[0078] (2) Preparation of N-(4-(5-amino-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide 1h:

[0079] 5-amino-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 1f (100 mg, 0.28 mmol), potassium trifluoro-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]borane 1g (82 mg, 0.28 mmol), cesium carbonate (182 mg, 0.56 mmol) and X-Phos (13 mg, 0.028 mmol) were suspended in THF (5 mL) and water (0.5 mL), and then palladium acetate (3 mg, 0.014 mmol) was added under nitrogen protection. The mixture was then heated to 80°C for 16 hours. After the reaction was completed, it was filtered. The filtrate was concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave N-(4-(5-amino-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide (100 mg, yield: 77%) as a pale yellow solid. MS-ESI (m / z): 465 [M+1] +

[0080] Step 5: Preparation of 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide 1

[0081] N-(4-(5-amino-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide (100 mg, 0.21 mmol) was dissolved in sulfuric acid (112 μL, 2.1 mmol) and trifluoroacetic acid (643 μL, 8.4 mmol). The reaction mixture was heated to 55°C and stirred for 5 h. After the reaction was completed, the reaction mixture was poured into an ice-water mixture, neutralized with sodium bicarbonate, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. Further purification by silica gel flash column chromatography, eluting with 0-10% MeOH in DCM, gave 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide (80 mg, yield: 77%) as a white solid. ESI-MS m / z: 483 [M+H] + .

[0082] 1H-NMR(DMSO-d6,400MHz)δ:8.83(t,J=6.1Hz,1H),7.51(dd,J=9.3,3.3Hz,1H),7.48-7.39(m,4 H),7.33(ddd,J=9.0,7.9,3.3Hz,1H),7.18(dd,J=9.0,4.3Hz,1H),6.67(s,2H),5.35-5.22(m,l H), 4.55 (d, J = 6.1Hz, 2H), 1.61 (d, J = 6.7Hz, 3H).

[0083] Example 1: 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]deuterated methyl]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide (2)

[0084]

[0085] The synthesis method refers to the preparation of the above C1 compound, wherein in the ninth step, 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane is replaced by 2-(bromodeuterated methyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane. (30 mg, white solid) ESI-MS m / z: 485 [M+H] + .

[0086] 1 H-NMR(DMSO-d6,400MHz)δ:8.84(t,J=6.1Hz,l H),7.52(dd,J=9.3,3.3Hz,1H),7.49-7.40(m,4H),7.38-7.31(m,l H),7.19(dd,J=9.0,4.3Hz,1H),6.68(s,2H),5.34-5.24(m,1H),1.62(d,J=6.7Hz,3H).

[0087] Example 2: 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl-d]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide (3)

[0088]

[0089] Step 1: Preparation of 5-(di-tert-butyloxycarbonylamino)-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole-4-carbonitrile 3a

[0090] 5-Amino-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole (5 g, 14 mmol) was dissolved in tetrahydrofuran (50 mL), DMAP (1.7 g, 14 mmol) and Boc anhydride (7.63 g, 35 mmol) were added, and then heated to 70 ° C and stirred for 16 h. After the reaction was completed, it was concentrated under reduced pressure and further purified by silica gel flash column chromatography to obtain 5-(di-tert-butyloxycarbonylamino)-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 3a (6 g, yield: 77%) as a white solid. ESI-MS m / z: 559 [M+H] +

[0091] Step 2: Preparation of 5-(di-tert-butyloxycarbonylamino)-4-cyano-3-(4-formylphenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 3b

[0092] 5-(Di-tert-butyloxycarbonylamino)-4-cyano-3-(4-bromophenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 3a (6 g, 10.7 mmol) was dissolved in DMSO (50 mL), and PdCl2(Amphos)2 (710 mg, 1 mmol), DIPEA (4.1 g, 32.1 mmol), triethylsilane (2.4 g, 21.1 mmol), C O atmosphere, heated to 100 ° C for 2 hours. After the reaction, cooled to room temperature, diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride for the organic phase, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and further purified by silica gel flash column chromatography to obtain 5-(di-tert-butyloxycarbonylamino)-4-cyano-3-(4-formylphenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 3b (4 g, yield: 73%) as a white solid. ESI-MSm / z: 509 [M+H] +

[0093] Step 3: Preparation of 5-fluoro-2-deuterated methoxybenzamide 3c

[0094] 5-Fluoro-2-deuterated methoxybenzoyl chloride 1g-4 (1g, 5.2mmol) was dissolved in tetrahydrofuran (10mL), and slowly added dropwise to the vigorously stirred ammonia water under ice bath. After the addition, it was stirred at room temperature for 1h. After the reaction was completed, it was extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 5-fluoro-2-deuterated methoxybenzamide 3c (0.8g, yield: 88%) as a white solid. ESI-MS m / z: 173[M+H] +

[0095] Step 4: Preparation of N-(4-(5-(di-tert-butyloxycarbonylamino)-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)-methyl-d-phenyl)-5-fluoro-2-(deuterated methoxy)benzamide 3d

[0096] 5-(Di-tert-butoxycarbonylamino)-4-cyano-3-(4-formylphenyl)-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazole 3b (500 mg, 0.98 mmol) and 5-fluoro-2-deuterated methoxybenzamide 3c (169 mg, 0.98 mmol) were dissolved in methanol (5 mL), acetic acid (59 mg, 0.98 mmol) was added, and the mixture was stirred at room temperature for 1 h. Then sodium borodeuteride (41 mg, 0.98 mmol) was added in batches. After the addition was complete, the mixture was stirred at room temperature for 16 h. After the reaction, the mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and further purified by silica gel flash column chromatography to obtain N-(4-(5-(di-tert-butyloxycarbonylamino)-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)-methyl-d-phenyl)-5-fluoro-2-(deuterated methoxy)benzamide 3d (500 mg, yield: 76%) as a white solid. ESI-MS m / z: 666[M+H] +

[0097] Step 5: Preparation of 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl-d]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide 3

[0098] N-(4-(5-(di-tert-butyloxycarbonylamino)-4-cyano-1-(1,1,1-trifluoropropane-2-yl)-1H-pyrazol-3-yl)-methyl-d-phenyl)-5-fluoro-2-(deuterated methoxy)benzamide 3d (100 mg, 0.15 mmol) was dissolved in sulfuric acid (80 μL, 1.5 mmol) and trifluoroacetic acid (459 μL, 6 mmol). The reaction mixture was heated to 55 °C and stirred for 5 h. After the reaction was completed, the reaction mixture was poured into an ice-water mixture, neutralized with sodium bicarbonate, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave 5-amino-3-[4-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl-d]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide 3 (60 mg, yield: 83%) as a white solid. ESI-MS m / z: 484 [M+H] + .

[0099] 1 H-NMR(DMSO-d6,400MHz)δ:8.83(t,J=6.1Hz,1H),7.51(dd,J=9.3,3.3Hz,1H),7.48-7.39(m,4 H),7.33(ddd,J=9.0,7.9,3.3Hz,1H),7.18(dd,J=9.0,4.3Hz,1H),6.67(s,2H),5.35-5.22(m,l H), 4.55 (d, J = 6.1Hz, 1H), 1.61 (d, J = 6.7Hz, 3H).

[0100] Example 3: 5-amino-3-[4-[[(5-fluoro-2-deuteromethoxy-benzoyl)amino]methyl]phenyl]-1-(2,2,2-trifluoro-1-deuteromethyl-ethyl)pyrazole-4-carboxamide

[0101]

[0102] The synthesis method refers to the preparation of the above C1 compound, wherein in the third step (1,1,1-trifluoropropane-2-yl)hydrazine hydrochloride is replaced by (1,1,1-trifluoropropane-2-yl-3,3,3-D3)hydrazine hydrochloride, (51 mg, white solid) ESI-MS m / z: 486 [M+H] +

[0103] 1 H-NMR(DMSO-d6,400MHz)δ:8.83(t,J=6.1Hz,1H),7.51(dd,J=9.3,3.3Hz,1H),7.48-7.39(m,4 H),7.33(ddd,J=9.0,7.9,3.3Hz,1H),7.18(dd,J=9.0,4.3Hz,1H),6.67(s,2H),5.35-5.22(m,l H),4.55(d,J=6.1Hz,2H).

[0104] Referring to the above examples, the following similar compounds were synthesized

[0105]

[0106]

[0107]

[0108] Example 11: 2-(4-((5-fluoro-2-(deuterated methoxy)benzamido)methyl)phenyl)-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide

[0109]

[0110] Step 1: Preparation of ethyl 3-(5-amino-3-(4-bromophenyl)-4-cyano-1H-pyrazol-1-yl)-4,4,4-trifluorobutyrate Ib

[0111] Ethyl 4,4,4-trifluoro-3-hydrazinobutyrate hydrochloride Ia (10.6 g, 45 mmol) and triethylamine (11.5 g, 114 mmol) were dissolved in EtOH (200 mL) solution, 2-[(4-bromophenyl)-methoxy-methylene]malononitrile 1d (10 g, 38 mmol) was added, and then heated to 100 ° C and stirred for 8 h. After the reaction was completed, it was concentrated under reduced pressure. Saturated aqueous ammonium chloride solution and ethyl acetate were added to the residue, the organic layer was separated, and washed with saturated sodium chloride in turn, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave ethyl 3-(5-amino-3-(4-bromophenyl)-4-cyano-1H-pyrazol-1-yl)-4,4,4-trifluorobutyrate Ib (12 g, yield: 73%) as a white solid. ESI-MS m / z: 431 [M+H] +

[0112] Step 2: Preparation of 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-hydroxybutan-2-yl)-4-cyano-1H-pyrazole Ic

[0113] 3-(5-amino-3-(4-bromophenyl)-4-cyano-1H-pyrazol-1-yl)-4,4,4-trifluorobutyric acid ethyl ester Ib (12 g, 27.8 mmol) was dissolved in methanol (100 mL), sodium borohydride (1 g, 27.8 mmol) was added in batches under ice bath, and stirred at room temperature for 16 h after the addition. After the reaction was completed, it was concentrated under reduced pressure, and saturated aqueous ammonium chloride solution and ethyl acetate were added to the residue. The organic layer was separated and washed with saturated sodium chloride in turn, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. It was further purified by silica gel flash column chromatography to obtain 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-hydroxybutan-2-yl)-4-cyano-1H-pyrazole Ic (8 g, yield: 74%) as a white solid. ESI-MS m / z: 389 [M+H] +

[0114] Step 3: Preparation of 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-oxobutane-2-yl)-4-nitrile-1H-pyrazole Id

[0115] 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-hydroxybutan-2-yl)-4-cyano-1H-pyrazole Ic (8 g, 20.5 mmol) was dissolved in dichloromethane (100 mL), and Dess-martine oxidant (10.4 g, 24.6 mmol) was added. The reaction was allowed to react at room temperature for 6 h. After the reaction was completed, the mixture was filtered, and the filtrate was washed with saturated sodium bicarbonate and saturated sodium chloride in turn, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-oxobutan-2-yl)-4-cyano-1H-pyrazole Id (5.5 g, yield: 69%) as a white solid. ESI-MS m / z: 387 [M+H] +

[0116] Step 4: Preparation of 2-(4-bromophenyl)-7-(trifluoromethyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine Ie

[0117] 5-amino-3-(4-bromophenyl)-1-(1,1,1-trifluoro-4-oxobutane-2-yl)-4-nitrile-1H-pyrazole Id (5 g, 13 mmol) was dissolved in methanol (5 mL), acetic acid (780 mg, 13 mmol) was added, and the mixture was stirred at room temperature for 1 h. Then sodium cyanoborohydride (816 mg, 13 mmol) was added in batches. After the addition, the mixture was stirred at room temperature for 16 h. After the reaction was completed, the mixture was diluted with water, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and further purified by silica gel flash column chromatography to obtain 2-(4-bromophenyl)-7-(trifluoromethyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine Ie (3.5 g, yield: 73%) as a white solid. ESI-MS m / z: 371 [M+H] +

[0118] Step 5: Preparation of N-(4-(3-cyano-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide If

[0119] 2-(4-bromophenyl)-7-(trifluoromethyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine Ie (100 mg, 0.27 mmol), potassium trifluoro-[[(5-fluoro-2-deuterated methoxy-benzoyl)amino]methyl]borane 1 g (79 mg, 0.27 mmol), cesium carbonate (176 mg, 0.54 mmol) and X-Phos (13 mg, 0.027 mmol) were suspended in THF (5 mL) and water (0.5 mL), and then palladium acetate (3 mg, 0.014 mmol) was added under nitrogen protection. The mixture was then heated to 80°C for 16 h. After the reaction was completed, it was filtered. The filtrate was concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave N-(4-(3-cyano-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide If (80 mg, yield: 62%) as a light yellow solid. ESI-MS m / z: 477 [M+H] +

[0120] Step 6: 2-(4-((5-fluoro-2-(deuterated methoxy)benzamido)methyl)phenyl)-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide

[0121] N-(4-(3-cyano-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidin-2-yl)benzyl)-5-fluoro-2-(deuterated methoxy)benzamide If (80 mg, 0.17 mmol) was dissolved in sulfuric acid (80 μL, 1.5 mmol) and trifluoroacetic acid (459 μL, 6 mmol). The reaction mixture was heated to 55°C and stirred for 5 h. After the reaction was completed, the reaction mixture was poured into an ice-water mixture, neutralized with sodium bicarbonate, extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. Further purification by silica gel flash column chromatography gave 2-(4-((5-fluoro-2-(deuterated methoxy)benzamido)methyl)phenyl)-7-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine-3-carboxamide (40 mg, yield: 48%) as a white solid. ESI-MS m / z: 495 [M+H] + .

[0122] 1H-NMR(DMSO-d6,400MHz)δ:8.82(t,J=6.1Hz,1H),7.52(dd,J=9.3,3.3Hz,1H),7.47-7.38(m,4H),7.33(ddd,J=9.0,7.9,3.3Hz,1H),7 .18(dd,J=9.0,4.3Hz,1H),6.73(s,1H),6.66(s,2H),5.39-5.21(m,1H),4.55(d,J=6.1Hz,2H),3.36-2.94(m,2H),2.35-1.89(m,2H).

[0123] Intermediate A: 2-(4-bromophenyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine

[0124]

[0125] Reference Example 11, Preparation of Intermediate 12e, wherein 4,4,4-trifluoro-3-hydrazinobutyric acid ethyl ester hydrochloride is replaced by 3-hydrazinopropionic acid ethyl ester hydrochloride. White solid. ESI-MS m / z: 303 [M+H] + .

[0126] Intermediate B: 2-(4-bromophenyl)-7-(methyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine

[0127]

[0128] Synthesis method:

[0129]

[0130] Step 1: Preparation of tert-butyl 2-(3-hydroxypropyl)hydrazine-1-carboxylate B-2

[0131] Dissolve tert-butyl carbazate (5 g, 37.8 mmol) and 4-hydroxy-2-butanone (3.3 g, 37.8 mmol) in methanol (50 mL), add acetic acid (2.2 g, 37.8 mmol), and stir the reaction solution at room temperature for 3 hours. Then, add sodium cyanoborohydride (2.3 g, 37.8 mmol) in batches, and stir the reaction solution at room temperature overnight. After the reaction is completed, quench with water, extract with ethyl acetate, wash the organic phase with saturated sodium chloride, dry with anhydrous sodium sulfate and concentrate under reduced pressure to obtain tert-butyl 2-(3-hydroxypropyl)hydrazine-1-carboxylate B-2 (6 g, yield: 83%), a colorless liquid.

[0132] ESI-MS m / z:191[M+H] + .

[0133] Step 2: Preparation of 3-hydrazinobutanol hydrochloride B-3

[0134] Dissolve 2-(3-hydroxypropyl)hydrazine-1-carboxylic acid tert-butyl ester B-2 (6 g, 31.5 mmol) in HCl-dioxane (4 M, 50 mL) and stir at room temperature for 2 hours. After the reaction is completed, directly concentrate under reduced pressure to obtain 3-hydrazinobutanol hydrochloride B-3 (5 g, yield: 90%) as a white solid. ESI-MS m / z: 105 [M+H] + .

[0135] From the third step to the fifth step, reference can be made to the synthesis of Example 11.

[0136] White solid, ESI-MS m / z: 317[M+H] + .

[0137] Intermediate C: 2-(4-bromophenyl)-7-(deuterated methyl)-3-nitrile-4,5,6,7-tetrahydropyrazolo[1,5-a]pyrimidine

[0138] The synthesis method refers to the synthesis of intermediate B, in which 4-hydroxy-2-butanone is replaced by 4-hydroxybutane-2-one-1,1,1-d3, white solid. ESI-MS m / z: 320 [M+H] + .

[0139] Referring to the above examples, the following similar compounds were synthesized

[0140]

[0141]

[0142]

[0143]

[0144] Comparative Example:

[0145] 5-Amino-3-[4-[[(5-fluoro-2-methoxy-benzoyl)amino]methyl]phenyl]-1-(2,2,2-trifluoro-1-methyl-ethyl)pyrazole-4-carboxamide (C2)

[0146]

[0147] ESI-MS m / z:480[M+H] + .H 1 NMR(400MHz,DMS0-d6)δ:8.84(t,J=6.1Hz,lH),7.53(dd,

[0148] J=9.2,3.3Hz,l H),7.48-7.41(m,4H),7.37-7.33(m,1H),7.18(dd,J=9.2,4.3Hz,1H),6.67(s,2H),5.35-5.23(m,l H), 4.56 (d, J = 6.0Hz, 2H), 3.91 (s, 3H), 1.62 (d, J = 6.9Hz, 3H).

[0149] Example 17 BTK kinase assay

[0150] In an experiment based on time-resolved fluorescence resonance energy transfer, the inhibitory effect of the example compounds on BTK kinase activity was tested. Recombinant BTK was pre-incubated with each example compound at room temperature in an assay buffer containing 50 mM Tris pH 7.4, 10 mM MgCl2, 2 mM MnCl2, 0.1 mM EDTA, 1 mM DTT, 20 nM SEB, 0.1% BSA, 0.005% tween-20 for 1 hour. The reaction was initiated by adding ATP (at ATP Km concentration) and peptide substrate (Biotin-AVLESEEELYSSARQ-NH2). After incubation at room temperature for 1 hour, an equal volume of a stop solution containing 50 mM HEPES pH 7.0, 800 mM KF, 20 mM EDTA, 0.1% BSA, p-Tyr66 antibody linked to Eu cryptate, and streptavidin-labeled XL665 was added to terminate the reaction. The plate was incubated for another hour at room temperature before the TR-FRET signal was read on a BMG PHERAstar FS instrument (ex 337 nm, em 620 nm / 665 nm). The residual enzyme activity and the IC value of each compound were calculated based on the ratio of the fluorescence at 615 nm to the fluorescence at 665 nm. 50 The data were fitted by a four-parameter logistic equation using Graphpad Prism software.

[0151] Table 1 Specific IC50 values ​​of the compounds of the present invention and comparative example compounds

[0152]

[0153]

[0154] Embodiment 18

[0155] The stability of the compounds was evaluated using human liver microsomes.

[0156] Assay system: The metabolic stability of the compounds of the invention was tested using liver microsomes from a mixture of men and women with 1 mM NADPH. The samples were analyzed using a mass spectrometer. HRMS was used to determine the peak area response ratio (the peak area corresponding to the test compound or control divided by the peak area of ​​the analytical internal standard) without running a standard curve. In order to detect all possible metabolites, HRMS scans were performed over the appropriate m / z range.

[0157] Assay conditions: The assay was performed with one incubation (N=1). Test compounds were incubated at 37°C in buffer containing 0.5 mg / ml liver microsomal protein. Reactions were initiated by the addition of cofactors and samples were taken at 0, 1, 2, 4, 8, 16, 32, 48 hours. A positive control (5 μM testosterone) was incubated in parallel and samples were taken at 0, 1, 2, 4, 8, 16, 32, 48 hours.

[0158] Assay quality control: The control compound testosterone was run in parallel to confirm (liver) microsomal enzyme activity. After the final time point, fluorimetry was used to confirm the addition of NADPH to the reaction mixture. The T1 / 2 of the control met the acceptable internal standard.

[0159] Analytical methods:

[0160] Liquid chromatography column: Thermo BDS Hypersil C18 30X2.0mm, 3μm, with guard column MP, buffer: 25mM formic acid buffer, pH 3.5;

[0161] Aqueous phase (A): 90% water, 10% buffer;

[0162] Organic phase (B): 90% acetonitrile, 10% buffer;

[0163] Flow rate: 300 μl / min;

[0164] Autosampler: injection volume 10 μl;

[0165] The gradient program is shown in Table 2.

[0166] Table 2 Gradient program

[0167]

[0168]

[0169] By using human liver microsomes, Examples 15, 16, and 17 described in the present invention showed a metabolic half-life of more than 22 hours, which was higher than the metabolic half-life of 16 hours in the comparative example. The results show that the relatively long metabolic half-life makes them have the potential to reduce medical doses and expand dosing intervals.

[0170] Although the present invention has been described by the specific embodiments above, it should not be construed as being limited thereto; rather, the present invention encompasses the general aspects disclosed above. Various modifications and various embodiments are possible without departing from the spirit and scope of the present invention.

Claims

1. An aromatic amide compound having a structure as shown in general formula (I) or a pharmaceutically acceptable salt thereof, The compound is specifically selected from: 。 2. The aromatic amide compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The pharmaceutically acceptable salt is an inorganic salt or an organic salt. The inorganic salt is selected from hydrochloride, hydrobromide, hydroiodide, perchlorate, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic salt is selected from formates, acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, succinates, glutarates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, p-toluenesulfonates, and ascorbates.

3. A pharmaceutical composition comprising the aromatic amide compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2.

4. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition also includes a pharmaceutically acceptable carrier, which is selected from the group consisting of microspheres, nanoparticles and liposomes.

5. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition also includes pharmaceutical excipients, which are selected from the group consisting of solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, flocculants and deflocculating agents, filter aids, and release retardants.

6. The pharmaceutical composition according to claim 3, characterized in that The dosage forms of the pharmaceutical composition include injection, freeze-dried powder for injection, suspension, implant, embolic agent, capsule, tablet, pill and oral solution.

7. Use of the aromatic amide compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 in the preparation of a BTK kinase agent.

8. Use of the aromatic amide compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 in the preparation of a medicament for treating a disease regulated by BTK.

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