Inhibitor crystal form containing five-membered ring derivative as well as preparation method and application of inhibitor crystal form

Abstract

The invention relates to a five-membered ring derivative-containing inhibitor crystal form, and a preparation method and application thereof. Particularly, the invention relates to a crystal form containing a five-membered ring derivative inhibitor, a preparation method of the crystal form and a pharmaceutical composition containing a therapeutically effective amount of the crystal form. The invention also relates to application of the compound as a biological regulator in preparation of drugs for treating autoimmune diseases, chronic inflammatory diseases, acute inflammatory diseases, autoinflammatory diseases, atherosclerosis, diabetes, fibrotic diseases, metabolic diseases, cancers, tumors, leukemia and lymphoma.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a five-membered ring derivative inhibitor crystal form, a preparation method and application thereof. Background Art

[0002] MK2 (MAPK activated protein kinase 2) is a protein kinase that is an important downstream molecule of the classic inflammatory signal transduction pathway MAPK and regulates the expression of inflammatory factors. After cells are stimulated by inflammatory factors, the activated p38 MAPK protein phosphorylates and activates MK2, leading to the phosphorylation of MK2 substrate TTP, promoting the stability of pro-inflammatory cytokine mRNA, thereby increasing the protein expression level of inflammatory cytokines.

[0003] In both mouse arthritis models and chondrocytes of osteoarthritis patients, MK2 is activated. Inhibition of the p38-MK2 pathway by genetic or pharmacological means can inhibit the expression of inflammatory factors, indicating that MK2 is involved in mediating the inflammatory regulation of arthritis. p38-MK2 inhibitors are expected to treat patients with relapsed, refractory rheumatoid arthritis who have poor response to existing therapies.

[0004] Currently, only Aclaris Therapeutics' p38-MK2 inhibitor ATI-450 has entered clinical stage IIb for rheumatoid arthritis. Preliminary clinical trial results show that ATI-450 is safe and tolerable in patients with rheumatoid arthritis, with outstanding and sustained efficacy. Other MK2 inhibitors in clinical trials include BMS's CC-99677 and Moerae Matrix's MMI-0100, both of which can directly inhibit the activity of MK2, with a different mechanism from ATI-450.

[0005] As drugs, p38-MK2 inhibitors have good application prospects in the pharmaceutical industry. p38-MK2 inhibitors provide a safer and more effective treatment for patients with rheumatoid arthritis who have relapsed, are refractory, and respond poorly to existing therapies; p38-MK2 inhibitors have a broad spectrum of anti-inflammatory effects (TNF / IL-6 / IL-1), and can theoretically be used to treat a variety of inflammatory diseases and autoimmune diseases; p38-MK2 inhibitors are small molecule inhibitors, which have advantages over large molecule drugs such as TNF monoclonal antibodies, such as oral convenience and good patient compliance. At the same time, p38-MK2 inhibitors have not been reported to have the toxic side effects of JAK inhibitors such as infection, tumors, thrombosis, and cardiac toxicity. Therefore, there is a huge clinical demand for the development of new p38-MK2 inhibitors.

[0006] The international application (application number: PCT / CN2023 / 072393) discloses the structures of a series of heterocyclic derivative inhibitors. In subsequent research and development, in order to make the products easy to handle, filter, dry, store, have long-term stability, and have high bioavailability, the present invention has conducted a comprehensive study on the crystal forms of the above-mentioned substances and is committed to obtaining the most suitable crystal form. Summary of the invention

[0007] All contents involved in patent application PCT / CN2023 / 072393 are added to the present invention by citation.

[0008] The object of the present invention is to provide a crystalline form of a compound represented by general formula (I) or a stereoisomer thereof

[0009]

[0010] in:

[0011] X1 is selected from N or CR 1 ;

[0012] R 1 is selected from hydrogen or halogen;

[0013] R 2 Selected from hydrogen, C 1-3 Alkyl or halogen;

[0014] R 3 Selected from C 1-3 Alkyl, C 1-3 Deuterated alkyl or C 3-6 Cycloalkyl;

[0015] R aa is selected from hydrogen or deuterium; and

[0016] R bb Selected from hydrogen or deuterium.

[0017] In a preferred embodiment of the invention, the compound is selected from the group consisting of

[0018] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0019] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0020] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0021] (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0022] (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0023] (S)-N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0024] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide;

[0025] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3;

[0026] (S)-N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide;

[0027] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide; or

[0028] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide.

[0029] In a preferred embodiment of the present invention, the crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide is crystalline form A, and the X-ray powder diffraction pattern of the crystalline form A has a diffraction peak at 2θ of 5.3±0.2°; or has a diffraction peak at 9.4±0.2°; or has a diffraction peak at 10.7±0.2°; or has a diffraction peak at 1 The invention has a diffraction peak at 2.3±0.2°; or has a diffraction peak at 15.9±0.2°; or has a diffraction peak at 16.8±0.2°; or has a diffraction peak at 17.4±0.2°; or has a diffraction peak at 19.1±0.2°; or has a diffraction peak at 22.2±0.2°; or has a diffraction peak at 26.8±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 of them.

[0030] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A comprises at least one or more diffraction peaks located at 2θ of 10.7±0.2°, 15.9±0.2°, and 17.4±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 5.3±0.2°, 9.4±0.2°, 12.3±0.2°, 19.1±0.2°, and 22.2±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0031] 10.7±0.2°, 15.9±0.2°;

[0032] 10.7±0.2°, 17.4±0.2°;

[0033] 15.9±0.2°, 17.4±0.2°;

[0034] 10.7±0.2°, 15.9±0.2°, 17.4±0.2°;

[0035] 5.3±0.2°, 10.7±0.2°, 15.9±0.2°;

[0036] 5.3±0.2°, 15.9±0.2°, 17.4±0.2°;

[0037] 5.3±0.2°, 10.7±0.2°, 15.9±0.2°, 17.4±0.2°;

[0038] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 15.9±0.2°;

[0039] 5.3±0.2°, 9.4±0.2°, 15.9±0.2°, 17.4±0.2°;

[0040] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 15.9±0.2°, 17.4±0.2°;

[0041] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°;

[0042] 5.3±0.2°, 9.4±0.2°, 12.3±0.2°, 15.9±0.2°, 17.4±0.2°.

[0043] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A optionally further comprises one or more diffraction peaks located at 2θ of 13.4±0.2°, 13.9±0.2°, 16.8±0.2°, 18.8±0.2°, 22.6±0.2°, 26.8±0.2°, 28.6±0.2°; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, or 7 thereof; for example,

[0044] 16.8±0.2°, 26.8±0.2°;

[0045] 16.8±0.2°, 18.8±0.2°;

[0046] 16.8±0.2°, 18.8±0.2°, 26.8±0.2°;

[0047] 13.4±0.2°, 16.8±0.2°, 26.8±0.2°;

[0048] 13.4±0.2°, 16.8±0.2°, 18.8±0.2°, 26.8±0.2°;

[0049] 13.4±0.2°, 16.8±0.2°, 22.6±0.2°, 26.8±0.2°;

[0050] 13.4±0.2°, 16.8±0.2°, 18.8±0.2°, 22.6±0.2°, 26.8±0.2°;

[0051] 13.4±0.2°, 13.9±0.2°, 16.8±0.2°, 22.6±0.2°, 26.8±0.2°;

[0052] 13.4±0.2°, 13.9±0.2°, 16.8±0.2°, 18.8±0.2°, 22.6±0.2°, 26.8±0.2°;

[0053] 13.4±0.2°, 16.8±0.2°, 18.8±0.2°, 22.6±0.2°, 26.8±0.2°, 28.6±0.2°.

[0054] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A comprises one or more diffraction peaks located at 2θ of 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 13.4±0.2°, 15.9±0.2°, 16.8±0.2°, 17.4±0.2°, 18.8±0.2°, 19.1±0.2°, 22.2±0.2°, and 26.8±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 selected therefrom; for example, the X-ray powder diffraction pattern of the crystalline form A has diffraction peaks at the following positions at 2θ:

[0055] 9.4±0.2°, 10.7±0.2°, 15.9±0.2°, 17.4±0.2°;

[0056] 10.7±0.2°, 15.9±0.2°, 17.4±0.2°, 19.1±0.2°;

[0057] 5.3±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°, 17.4±0.2°;

[0058] 5.3±0.2°, 10.7±0.2°, 22.2±0.2°, 15.9±0.2°, 17.4±0.2°;

[0059] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 15.9±0.2°, 17.4±0.2°, 19.1±0.2°;

[0060] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°, 17.4±0.2°;

[0061] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°, 17.4±0.2°, 19.1±0.2°, 22.2±0.2°;

[0062] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°, 16.8±0.2°, 17.4±0.2°, 19.1±0.2°;

[0063] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 15.9±0.2°, 16.8±0.2°, 17.4±0.2°, 19.1±0.2°, 22.2±0.2°, 26.8±0.2°;

[0064] 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 13.4±0.2°, 15.9±0.2°, 17.4±0.2°, 18.8±0.2°, 19.1±0.2°, 22.2±0.2°.

[0065] In a preferred embodiment of the present invention, the crystal form A uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks represented by 2θ angles and interplanar spacing d values ​​are shown in Table 1.

[0066] Table 1

[0067]

[0068]

[0069] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A is substantially as follows Figure 1 shown.

[0070] In a preferred embodiment of the present invention, the DSC spectrum of the crystalline form A is as follows Figure 2 shown.

[0071] In a preferred embodiment of the present invention, the crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide is crystalline form B, and the X-ray powder diffraction pattern of crystalline form B has a diffraction peak at 2θ of 5.2±0.2°; or has a diffraction peak at 8.0±0.2°; or has a diffraction peak at 10.1±0.2°; or has a diffraction peak at 1 It has a diffraction peak at 0.4±0.2°; or a diffraction peak at 11.4±0.2°; or a diffraction peak at 12.0±0.2°; or a diffraction peak at 12.7±0.2°; or a diffraction peak at 15.5±0.2°; or a diffraction peak at 17.8±0.2°; or a diffraction peak at 19.1±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 of them.

[0072] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form B comprises at least one or more diffraction peaks located at 2θ of 10.4±0.2°, 11.4±0.2°, and 15.5±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 12.0±0.2°, and 17.8±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0073] 10.4±0.2°, 11.4±0.2°;

[0074] 10.4±0.2°, 15.5±0.2°;

[0075] 11.4±0.2°, 15.5±0.2°;

[0076] 10.4±0.2°, 11.4±0.2°, 15.5±0.2°;

[0077] 10.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0078] 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0079] 10.4±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0080] 8.0±0.2°, 10.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0081] 8.0±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0082] 8.0±0.2°, 10.4±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0083] 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0084] 8.0±0.2°, 10.1±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°.

[0085] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form B optionally further comprises one or more diffraction peaks located at 2θ of 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°, 21.7±0.2°, and 25.1±0.2°; preferably, at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, or 7 thereof; for example,

[0086] 12.7±0.2°, 19.1±0.2°;

[0087] 16.9±0.2°, 19.1±0.2°;

[0088] 12.7±0.2°, 16.9±0.2°, 19.1±0.2°;

[0089] 12.2±0.2°, 12.7±0.2°, 19.1±0.2°;

[0090] 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 19.1±0.2°;

[0091] 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°;

[0092] 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°;

[0093] 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 19.1±0.2°, 25.1±0.2°;

[0094] 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°, 25.1±0.2°;

[0095] 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°, 21.7±0.2°.

[0096] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form B comprises one or more diffraction peaks located at 2θ of 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 12.2±0.2°, 12.7±0.2°, 15.5±0.2°, 16.9±0.2°, 17.8±0.2°, and 19.1±0.2°; preferably, it comprises 4, 5, 6, 8, 10, or 12 selected therefrom; for example, the X-ray powder diffraction pattern of the crystalline form B has diffraction peaks at the following positions at 2θ:

[0097] 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 15.5±0.2°;

[0098] 8.0±0.2°, 10.4±0.2°, 11.4±0.2°, 15.5±0.2°;

[0099] 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0100] 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 15.5±0.2°, 17.8±0.2°;

[0101] 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 15.5±0.2°, 17.8±0.2°;

[0102] 8.0±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 15.5±0.2°, 17.8±0.2°;

[0103] 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 15.5±0.2°, 17.8±0.2°;

[0104] 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 15.5±0.2°, 17.8±0.2°, 19.1±0.2°;

[0105] 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 12.7±0.2°, 15.5±0.2°, 17.8±0.2°, 19.1±0.2°;

[0106] 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 12.2±0.2°, 15.5±0.2°, 16.9±0.2°, 17.8±0.2°.

[0107] In a preferred embodiment of the present invention, the crystal form B uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks represented by 2θ angles and interplanar spacing d values ​​are shown in Table 2.

[0108] Table 2

[0109]

[0110]

[0111] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form B is substantially as follows Figure 3 shown.

[0112] In a preferred embodiment of the present invention, the crystalline form B has Figure 4 DSC spectrum shown.

[0113] In a preferred embodiment of the present invention, the crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propane-2-yl)acetamide is crystalline form C, and the X-ray powder diffraction pattern of the crystalline form C has a diffraction peak at 2θ of 7.3±0.2°; or has a diffraction peak at 8.6±0.2°; or has a diffraction peak at 9.2±0.2°; or has a diffraction peak at 11 .7±0.2°; or has a diffraction peak at 15.2±0.2°; or has a diffraction peak at 17.2±0.2°; or has a diffraction peak at 19.8±0.2°; or has a diffraction peak at 21.6±0.2°; or has a diffraction peak at 22.3±0.2°; or has a diffraction peak at 24.2±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 of them.

[0114] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form C comprises at least one or more diffraction peaks located at 2θ of 7.3±0.2°, 8.6±0.2°, and 15.2±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 9.2±0.2°, 11.7±0.2°, 21.6±0.2°, 22.3±0.2°, and 24.2±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0115] 7.3±0.2°, 8.6±0.2°;

[0116] 8.6±0.2°, 15.2±0.2°;

[0117] 7.3±0.2°, 15.2±0.2°;

[0118] 7.3±0.2°, 8.6±0.2°, 15.2±0.2°;

[0119] 8.6±0.2°, 9.2±0.2°, 15.2±0.2°;

[0120] 7.3±0.2°, 9.2±0.2°, 15.2±0.2°;

[0121] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 15.2±0.2°;

[0122] 8.6±0.2°, 9.2±0.2, 15.2±0.2°, 24.2±0.2°;

[0123] 7.3±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°;

[0124] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 15.2±0.2°, 24.2±0.2°;

[0125] 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 24.2±0.2°;

[0126] 7.3±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 24.2±0.2°.

[0127] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form C optionally further comprises one or more diffraction peaks located at 2θ of 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.6±0.2°, 27.8±0.2°, and 37.0±0.2°; preferably, at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, or 7 thereof; for example,

[0128] 17.2±0.2°, 19.8±0.2°;

[0129] 18.5±0.2°, 19.8±0.2°;

[0130] 17.2±0.2°, 18.5±0.2°, 19.8±0.2°;

[0131] 17.2±0.2°, 19.8±0.2°, 27.8±0.2°;

[0132] 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.8±0.2°;

[0133] 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°;

[0134] 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.8±0.2°;

[0135] 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.8±0.2°, 37.0±0.2°;

[0136] 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.8±0.2°, 37.0±0.2°;

[0137] 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.6±0.2°, 27.8±0.2°.

[0138] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form C comprises one or more diffraction peaks located at 2θ of 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 21.6±0.2°, 22.3±0.2°, 24.2±0.2°, and 27.8±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 selected therefrom; for example, the X-ray powder diffraction pattern of the crystalline form C has diffraction peaks at the following positions at 2θ:

[0139] 7.3±0.2°, 8.6±0.2°, 15.2±0.2°, 24.2±0.2°;

[0140] 7.3±0.2°, 8.6±0.2°, 11.7±0.2°, 15.2±0.2°;

[0141] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 15.2±0.2°, 22.3±0.2°;

[0142] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 15.2±0.2°, 21.6±0.2°;

[0143] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 24.2±0.2°;

[0144] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 15.2±0.2°, 22.3±0.2°, 24.2±0.2°;

[0145] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 21.6±0.2°, 22.3±0.2°, 24.2±0.2°;

[0146] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 17.2±0.2°, 19.8±0.2°, 24.2±0.2°;

[0147] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 17.2±0.2°, 19.8±0.2°, 21.6±0.2°, 22.3±0.2°, 24.2±0.2°;

[0148] 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 18.5±0.2°, 21.6±0.2°, 22.3±0.2°, 24.2±0.2°, 27.8±0.2°.

[0149] In a preferred embodiment of the present invention, the crystal form C uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks represented by 2θ angles and interplanar spacing d values ​​are shown in Table 3.

[0150] Table 3

[0151]

[0152]

[0153] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form C is substantially as follows Figure 5 shown.

[0154] In a preferred embodiment of the present invention, the crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide is crystalline form D, and the X-ray powder diffraction pattern of the crystalline form D has a diffraction peak at 2θ of 7.2±0.2°; or has a diffraction peak at 8.6±0.2°; or has a diffraction peak at 9.3±0.2°; or has a diffraction peak at 11 .8±0.2°; or has a diffraction peak at 15.2±0.2°; or has a diffraction peak at 15.6±0.2°; or has a diffraction peak at 18.7±0.2°; or has a diffraction peak at 20.1±0.2°; or has a diffraction peak at 21.5±0.2°; or has a diffraction peak at 24.5±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 of them.

[0155] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form D comprises at least one or more diffraction peaks located at 2θ of 7.2±0.2°, 9.3±0.2°, and 15.2±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 8.6±0.2°, 11.8±0.2°, 18.7±0.2°, 20.1±0.2°, and 24.5±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0156] 7.2±0.2°, 9.3±0.2°;

[0157] 9.3±0.2°, 15.2±0.2°;

[0158] 7.2±0.2°, 15.2±0.2°;

[0159] 7.2±0.2°, 9.3±0.2°, 15.2±0.2°;

[0160] 8.6±0.2°, 9.3±0.2°, 15.2±0.2°;

[0161] 7.2±0.2°, 8.6±0.2°, 15.2±0.2°;

[0162] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 15.2±0.2°;

[0163] 8.6±0.2°, 9.3±0.2°, 15.2±0.2°, 24.5±0.2°;

[0164] 7.2±0.2°, 8.6±0.2°, 15.2±0.2°, 24.5±0.2°;

[0165] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 15.2±0.2°, 24.5±0.2°;

[0166] 8.6±0.2°, 9.3±0.2°, 15.2±0.2°, 18.7±0.2°, 24.5±0.2°;

[0167] 7.2±0.2°, 8.6±0.2°, 15.2±0.2°, 18.7±0.2°, 24.5±0.2°.

[0168] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form D optionally further comprises one or more diffraction peaks located at 2θ of 12.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 23.7±0.2°, 25.4±0.2°, 28.2±0.2°; preferably at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, or 7 thereof; for example,

[0169] 15.6±0.2°, 21.5±0.2°;

[0170] 21.5±0.2°, 28.2±0.2°;

[0171] 15.6±0.2°, 21.5±0.2°, 28.2±0.2°;

[0172] 15.6±0.2°, 17.2±0.2°, 21.5±0.2°;

[0173] 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 28.2±0.2°;

[0174] 15.6±0.2°, 21.5±0.2°, 25.4±0.2°, 28.2±0.2°;

[0175] 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 25.4±0.2°, 28.2±0.2°;

[0176] 12.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 28.2±0.2°;

[0177] 12.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 25.4±0.2°, 28.2±0.2°;

[0178] 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 23.7±0.2°, 225.4±0.2°, 28.2±0.2°.

[0179] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form D comprises one or more diffraction peaks located at 2θ of 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 15.6±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 21.5±0.2°, 24.5±0.2°, and 28.2±0.2°; preferably, it comprises 4, 5, 6, 8, 10, or 12 selected therefrom; for example, the X-ray powder diffraction pattern of the crystalline form D has diffraction peaks at the following positions at 2θ:

[0180] 7.2±0.2°, 9.3±0.2°, 15.2±0.2°, 24.5±0.2°;

[0181] 7.2±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°;

[0182] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°;

[0183] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 15.2±0.2°, 18.7±0.2°;

[0184] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 24.5±0.2°;

[0185] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 15.2±0.2°, 18.7±0.2°, 24.5±0.2°;

[0186] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 18.7±0.2°, 20.1±0.2°, 24.5±0.2°;

[0187] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 15.6±0.2°, 21.5±0.2°, 24.5±0.2°;

[0188] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 15.6±0.2°, 18.7±0.2°, 20.1±0.2°, 21.5±0.2°, 24.5±0.2°;

[0189] 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 24.5±0.2°, 28.2±0.2°.

[0190] In a preferred embodiment of the present invention, the crystal form D uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks represented by 2θ angles and interplanar spacing d values ​​are shown in Table 4.

[0191] Table 4

[0192]

[0193]

[0194] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form D is substantially as follows Figure 6 shown.

[0195] In a preferred embodiment of the present invention, the crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide is crystalline form E, and the X-ray powder diffraction pattern of the crystalline form E has a diffraction peak at 2θ of 4.4±0.2°; or has a diffraction peak at 10.8±0.2°; or has a diffraction peak at 11.6±0.2°; or has a diffraction peak at 1 The invention has a diffraction peak at 2.6±0.2°; or has a diffraction peak at 13.0±0.2°; or has a diffraction peak at 13.3±0.2°; or has a diffraction peak at 15.3±0.2°; or has a diffraction peak at 16.9±0.2°; or has a diffraction peak at 21.0±0.2°; or has a diffraction peak at 25.4±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 thereof.

[0196] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form E comprises at least one or more diffraction peaks located at 2θ of 4.4±0.2°, 13.3±0.2°, and 16.9±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 15.3±0.2°, and 25.4±0.2°, preferably 2, 3, 4 or 5 of them; for example,

[0197] 4.4±0.2°, 13.3±0.2°;

[0198] 13.3±0.2°, 16.9±0.2°;

[0199] 4.4±0.2°, 16.9±0.2°;

[0200] 4.4±0.2°, 13.3±0.2°, 16.9±0.2°;

[0201] 10.8±0.2°, 13.3±0.2°, 16.9±0.2°;

[0202] 4.4±0.2°, 10.8±0.2°, 16.9±0.2°;

[0203] 4.4±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2°;

[0204] 10.8±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2°;

[0205] 4.4±0.2°, 10.8±0.2°, 16.9±0.2°, 25.4±0.2;

[0206] 4.4±0.2°, 10.8±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0207] 10.8±0.2°, 12.6±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0208] 4.4±0.2°, 10.8±0.2°, 12.6±0.2°, 16.9±0.2°, 25.4±0.2.

[0209] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form E optionally further comprises one or more diffraction peaks located at 2θ of 13.0±0.2°, 14.2±0.2°, 17.5±0.2°, 18.7±0.2°, 21.0±0.2°, 21.9±0.2°, and 26.2±0.2°; preferably, at least any 2-3, or 4-5, or 6-7 thereof; further preferably, any 2, 3, 4, 5, 6, or 7 thereof; for example,

[0210] 13.0±0.2°, 21.0±0.2°;

[0211] 21.0±0.2°, 21.9±0.2°;

[0212] 13.0±0.2°, 21.0±0.2°, 21.9±0.2°;

[0213] 13.0±0.2°, 14.2±0.2°, 21.0±0.2°;

[0214] 13.0±0.2°, 14.2±0.2°, 21.0±0.2°, 21.9±0.2°;

[0215] 13.0±0.2°, 17.5±0.2°, 21.0±0.2°, 21.9±0.2°;

[0216] 13.0±0.2°, 14.2±0.2°, 17.5±0.2°, 21.0±0.2°, 21.9±0.2°;

[0217] 13.0±0.2°, 14.2±0.2°, 18.7±0.2°, 21.0±0.2°, 21.9±0.2°;

[0218] 13.0±0.2°, 14.2±0.2°, 17.5±0.2°, 18.7±0.2°, 21.0±0.2°, 21.9±0.2°;

[0219] 13.0±0.2°, 14.2±0.2°, 17.5±0.2°, 18.7±0.2°, 21.0±0.2°, 21.9±0.2°.

[0220] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form E comprises one or more diffraction peaks located at 2θ of 4.4±0.2°, 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 13.0±0.2°, 13.3±0.2°, 14.2±0.2°, 15.3±0.2°, 16.9±0.2°, 21.0±0.2°, 21.9±0.2°, and 25.4±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 selected therefrom; for example, the X-ray powder diffraction pattern of the crystalline form E has diffraction peaks at the following positions at 2θ:

[0221] 4.4±0.2°, 10.8±0.2°, 13.3±0.2°, 16.9±0.2°;

[0222] 4.4±0.2°, 12.6±0.2°, 13.3±0.2°, 16.9±0.2°;

[0223] 4.4±0.2°, 12.6±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0224] 4.4±0.2°, 13.3±0.2°, 15.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0225] 4.4±0.2°, 10.8±0.2°, 12.6±0.2°, 13.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0226] 4.4±0.2°, 10.8±0.2°, 13.3±0.2°, 15.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0227] 4.4±0.2°, 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 13.3±0.2°, 15.3±0.2°, 16.9±0.2°, 25.4±0.2;

[0228] 4.4±0.2°, 10.8±0.2°, 12.6±0.2°, 13.0±0.2°, 13.3±0.2°, 16.9±0.2°, 21.0±0.2°, 25.4±0.2;

[0229] 4.4±0.2°, 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 13.0±0.2°, 13.3±0.2°, 15.3±0.2°, 16.9±0.2°, 21.0±0.2°, 25.4±0.2;

[0230] 4.4±0.2°, 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 13.3±0.2°, 14.2±0.2°, 15.3±0.2°, 16.9±0.2°, 21.9±0.2°, 25.4±0.2.

[0231] In a preferred embodiment of the present invention, the crystal form E uses Cu-Kα radiation, and the X-ray characteristic diffraction peaks represented by 2θ angles and interplanar spacing d values ​​are shown in Table 5.

[0232] Table 5

[0233]

[0234] In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form E is substantially as follows Figure 7 shown.

[0235] In a preferred embodiment of the present invention, the 2θ error between the top ten diffraction peak positions with relative peak intensity in the X-ray powder diffraction pattern of the crystalline form AE and the diffraction peak at the corresponding position in the accompanying drawing is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°.

[0236] The present invention also provides a method for preparing the above-mentioned compound or a crystalline form of its stereoisomer, which is method one or method two:

[0237] Method 1 comprises the following steps: the compound is slurried with a solvent at a certain temperature to obtain a crystalline form, preferably, the temperature is 0 to 50°C; more preferably, the temperature is 20 to 30°C;

[0238] The solvent is selected from one or more of water, methanol, ethanol, isopropyl acetate, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol n-propanol, tert-butanol, 2-butanone, 3-pentanone and N-methylpyrrolidone; preferably one or more of water, methanol, ethanol, isopropyl acetate, acetone, ethyl acetate, acetonitrile, dichloromethane, 1,4-dioxane, isopropanol, 2-butanone or 3-pentanone;

[0239] Method 2 comprises the following steps: dissolving the compound in a good solvent, adding a poor solvent and stirring until a solid is precipitated to obtain a crystalline form;

[0240] The good solvent is selected from one or more of ethyl acetate, isopropyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran or 1,4-dioxane; preferably one or more of acetone, dichloromethane or acetonitrile;

[0241] The poor solvent is selected from one or more of n-heptane, cyclohexane, n-hexane, n-pentane, water, ethyl acetate, methyl tert-butyl ether, toluene or isopropyl ether; preferably one or more of water, n-hexane or methyl tert-butyl ether.

[0242] The present invention also provides a pharmaceutical composition, which contains a therapeutically effective amount of the crystalline form of the above compound and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0243] The pharmaceutical composition of the invention includes suitable administration routes such as oral administration and injection, preferably oral administration route. The dosage form includes tablets, capsules, dispersions and suspensions, preferably tablets.

[0244] The present invention further relates to a pharmaceutical composition, which contains a therapeutically effective amount of a crystalline form of the above-mentioned compound; further, the weight percentage of the crystalline form of the compound in the composition is 0.1% to 95%, preferably 0.5% to 85%, more preferably 1% to 60%, further preferably 10% to 50%, further preferably 15-40%, further preferably 20-30%, further preferably 20-25% (based on the total weight of the pharmaceutical composition).

[0245] The present invention further relates to the crystal form of the compound, or the use of the pharmaceutical composition in preparing drugs for treating diseases mediated by p38 kinase.

[0246] The present invention further relates to the use of the crystal form of the compound or its pharmaceutical composition in the preparation of drugs for treating autoimmune diseases, metabolic diseases and tumors, etc. The autoimmune diseases are selected from chronic inflammatory diseases, acute inflammatory diseases, autoinflammatory diseases and atherosclerosis, etc.; the metabolic diseases are selected from diabetes and fibrotic diseases, etc.; the tumors are selected from leukemia and lymphoma, etc.

[0247] The present invention also relates to a method for treating, preventing and / or treating autoimmune diseases, metabolic diseases and tumors, which comprises administering to a patient a therapeutically effective dose of a crystalline form of the compound of the present invention, or a pharmaceutical composition thereof; wherein the autoimmune disease is selected from chronic inflammatory diseases, acute inflammatory diseases, autoinflammatory diseases and atherosclerosis, etc.; the metabolic disease is selected from diabetes and fibrotic diseases, etc.; the tumor is selected from leukemia and lymphoma, etc.

[0248] The present invention also provides methods of using the compounds or pharmaceutical compositions of the present invention to treat disease conditions, including but not limited to conditions associated with p38 kinase-mediated diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0249] Figure 1 This is an XRPD diagram of free base Form A.

[0250] Figure 2 This is a DSC diagram of free base form A.

[0251] Figure 3 This is the XRPD diagram of free base Form B.

[0252] Figure 4 This is the DSC diagram of free base form B.

[0253] Figure 5 This is an XRPD diagram of free base Form C.

[0254] Figure 6 This is the XRPD diagram of free base Form D.

[0255] Figure 7 This is an XRPD diagram of free base Form E.

[0256] Detailed description of the invention

[0257] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0258] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 8 carbon atoms, further preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 4-heptyl, 1-propylbutyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2-Methyl-2-ethyl-pentyl, 2-methyl-3-ethyl-pentyl, n-nonyl, 2-methyl-2-ethyl-hexyl, 2-methyl-3-ethyl-hexyl, 2,2-diethyl-pentyl, n-decyl, 3,3-diethyl-hexyl, 2,2-diethyl-hexyl, and various branched-chain isomers thereof. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, 4-heptyl, 1-propylbutyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available attachment point. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred in the present invention.

[0259] The term "alkylene" refers to a divalent alkyl group formed by further replacing one of the hydrogen atoms of an alkyl group, wherein alkyl is as defined above. For example, "methylene" refers to -CH2-, "ethylene" refers to -(CH2)2-, "propylene" refers to -(CH2)3-, "butylene" refers to -(CH2)4-, etc. The points of attachment of the alkylene chain to the rest of the molecule and to the radical group can be through one carbon or any two carbons within the chain. The alkylene group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available attachment point. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred in the present invention.

[0260] The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, which is a straight or branched group comprising 2 to 20 carbon atoms, preferably an alkenyl group containing 2 to 12 carbon atoms, more preferably an alkenyl group containing 2 to 8 carbon atoms, further preferably an alkenyl group containing 2 to 6 carbon atoms, and most preferably an alkenyl group containing 2 to 4 carbon atoms. For example, vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl or 3-butenyl, etc. Alkenyl can be substituted or unsubstituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0261] The term "alkenylene" refers to a divalent alkenyl group formed by further replacing one hydrogen atom of an alkenyl group, wherein the alkenyl group is as defined above, for example, vinylene, propenylene, butenylene, etc. The alkenylene group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0262] The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, which is a straight or branched group containing 2 to 20 carbon atoms, preferably an alkynyl group containing 2 to 12 carbon atoms, more preferably an alkynyl group containing 2 to 8 carbon atoms, further preferably an alkynyl group containing 2 to 6 carbon atoms, and most preferably an alkynyl group containing 2 to 4 carbon atoms. For example, ethynyl, propynyl, 1-butynyl, 2-butynyl or 3-butynyl, etc. Alkynyl can be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0263] The term "alkynylene" refers to a divalent alkynyl group formed by further replacing one of the hydrogen atoms of an alkynyl group, wherein the alkynyl group is as defined above. For example, ethynylene, propynylene, butynylene, etc. Alkynylidene can be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0264] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 carbon atoms, and further preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include cycloalkyls of spirocyclic, condensed and bridged rings, preferably cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl and cycloheptyl.

[0265] "Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples of haloalkyl groups include: trifluoromethyl, -CH2CF3,

[0266] "Haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0267] "Hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0268] The term "alkenylcarbonyl" refers to -C(O)-(alkenyl), wherein the definition of alkenyl is as described above. Non-limiting examples of alkenylcarbonyl include: vinylcarbonyl, propenylcarbonyl, butenylcarbonyl. Alkenylcarbonyl can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0269] "Aminocarbonyl" refers to NH2-C(O)-.

[0270] "Alkylaminocarbonyl" refers to an aminocarbonyl (NH2-C(O)-) group in which one or both of the two hydrogen atoms are replaced by alkyl groups, wherein alkyl is as defined above.

[0271] "Alkylamino" refers to an amino group in which one or both of the two hydrogen atoms are replaced by an alkyl group, wherein the alkyl group is as defined above.

[0272] "Alkylcarbonyl" or "acyl" refers to (alkyl)-C(O)- wherein alkyl is as previously described.

[0273] Different expressions such as “X is selected from A, B, or C”, “X is selected from A, B and C”, “X is A, B or C”, “X is A, B and C” all express the same meaning, that is, X can be any one or more of A, B, C.

[0274] The hydrogen atoms described in the present invention can be replaced by their isotope deuterium, and any hydrogen atom in the example compounds of the present invention can also be replaced by a deuterium atom.

[0275] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0276] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

[0277] "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0278] "Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0279] X-ray powder diffraction pattern (XRPD) refers to the experimentally observed diffraction pattern or the parameters derived therefrom, and the X-ray powder diffraction pattern is characterized by the peak position (abscissa) and the peak intensity (ordinate). It will be understood by those skilled in the art that the experimental error depends on the conditions of the instrument, the preparation of the sample and the purity of the sample. In particular, it is well known to those skilled in the art that the X-ray diffraction pattern will usually change with the conditions of the instrument, and those skilled in the art should understand that the appropriate error tolerance of XRPD can be: 2θ±0.5°; 2θ±0.4°; 2θ±0.3°; 2θ±0.2°. It should be particularly noted that the relative intensity of the X-ray diffraction pattern may also change with changes in experimental conditions, so the order of peak intensity cannot be used as the only or decisive factor. In addition, due to the influence of experimental factors such as sample height, the overall deviation of the peak angle will be caused, and a certain deviation is usually allowed. Therefore, it will be understood by those skilled in the art that any crystal form having the same or similar characteristic peaks as the spectrum of the present invention belongs to the scope of the present invention.

[0280] "TGA" refers to a thermogravimetric analysis (TGA) experiment.

[0281] "DSC" refers to a differential scanning calorimetry (DSC) experiment. DETAILED DESCRIPTION

[0282] The present invention is further described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0283] Example

[0284] The structure of the compound of the present invention is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS). The NMR chemical shift (δ) is given in parts per million (ppm). The NMR is measured using a Bruker AVANCE-400 nuclear magnetic spectrometer, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), deuterated chloroform (CDCl3) or deuterated water (D2O), and the internal standard is tetramethylsilane (TMS).

[0285] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1200 Infinity Series mass spectrometer, and HPLC was performed using an Agilent 1200DAD high pressure liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150×4.6 mm column).

[0286] Thin layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications used for TLC are 0.15mm-0.20mm, and the specifications used for thin layer chromatography separation and purification products are 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0287] The starting materials in the examples of the present invention are known and can be purchased on the market, or can be synthesized by or according to methods known in the art.

[0288] Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring in a dry nitrogen or argon atmosphere, the solvent is a dry solvent, and the reaction temperature is in degrees Celsius.

[0289] The eluent system of silica gel column chromatography and the developing solvent system of thin layer chromatography used for the intermediates and purified compounds in the examples include: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: dichloromethane and acetone system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

[0290] Unless otherwise specified, in the embodiments of the present invention, the ratios in the mobile phases in the HPLC chiral separation conditions and HPLC chiral analysis conditions are volume ratios.

[0291] Intermediate 1

[0292] 2-(1H-Pyrazol-3-yl)propan-2-ol

[0293]

[0294] first step

[0295] 1H-pyrazole-3-carboxylic acid methyl ester 1A (2g, 15.9mmol) was dissolved in tetrahydrofuran (50mL) and cooled to 0°C. Methylmagnesium chloride (1M, 47.58mL) was added dropwise to the reaction solution under stirring. After the addition was completed, the reaction was warmed to room temperature and stirred for 2 hours. Ethyl acetate (100mL) was added to the reaction solution, and the organic phase was washed with water and saturated ammonium chloride, dried, and concentrated to obtain the product 2-(1H-pyrazol-3-yl)propan-2-ol intermediate 1 (840mg), with a yield of 42%.

[0296] MS m / z(ESI):127.1[M+H] + .

[0297] 1 H NMR (400MHz, CDCl3) δ7.51(d,1H),6.64(s,2H),6.18(d,1H),1.62(s,6H).

[0298] Intermediate 2

[0299] 2'-Bromo-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one

[0300]

[0301] first step

[0302] 2-Bromo-5-methyl-pyridin-4-amine 2A (2.03 g, 10.86 mmol) and 2,2-dimethyl-6-(2-carbonylpropyl)-4H-1,3-dioxin-4-one (2 g, 10.86 mmol) were dissolved in 1,4-dioxane (80 mL) and stirred. The reaction was placed in a reactor preheated to 90°C and stirred for 3 hours. Concentrated sulfuric acid (1.8 g, 18.35 mmol) was added to the reaction solution and continued to stir for 1 hour. The reaction solution was cooled, the supernatant was removed, and water was added to the residual oil. The mixture was stirred at room temperature for 30 minutes and filtered to obtain 2'-bromo-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridine]-2-one 2B (1.7 g), with a yield of 53%.

[0303] MS m / z(ESI):296.1[M+H] + .

[0304] Step 2

[0305] 2'-Bromo-4-hydroxy-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 2B (1.7 g, 5.76 mmol) was dissolved in isopropanol (40 mL) and 1,2-dichloroethane (60 mL) and heated to 60°C with stirring. N-chlorosuccinamide (923 mg, 6.91 mmol) was added to the reaction solution in batches and stirring was continued for 2 hours. The reaction solution was concentrated, isopropanol (20 mL) was added to the residue, and the mixture was heated to 60°C and stirred for 0.5 hours. The reaction solution was cooled and filtered to obtain 1-(2-bromo-5-methyl-4-pyridyl)-3-chloro-4-hydroxy-6-methyl-pyridin-2-one 2C (1.71 g), with a yield of 90.1%.

[0306] MS m / z(ESI):330.9[M+H] + .

[0307] Step 3

[0308] 1-(2-bromo-5-methyl-4-pyridyl)-3-chloro-4-hydroxy-6-methyl-pyridin-2-one 2C (150 mg, 0.455 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (139 mg, 0.910 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred. 2-(Chloromethyl)-3,5-difluoro-pyridine (89 mg, 0.546 mmol) was added dropwise to the reaction solution, and heated to 80°C and stirred for 2 hours. Ethyl acetate (20 mL) was added to the reaction solution, and the organic phase was washed with water, saturated ammonium chloride and saturated sodium chloride, dried and concentrated. The residue was separated by silica gel column chromatography (eluent system A) to give 2'-bromo-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2H-[1,4'-bipyridinyl]-2-one intermediate 2 (140 mg) in a yield of 67.4%.

[0309] MS m / z(ESI):458.0[M+H] + .

[0310] Intermediate 3

[0311] 1-(Chloromethyl-d2)-2,4-difluorobenzene

[0312]

[0313] first step

[0314] Methyl 2,4-difluorobenzoate 3A (0.5 g, 2.90 mmol) was dissolved in tetrahydrofuran (8 mL) and stirred. A solution of lithium aluminum deuteride (122 mg, 2.90 mmol) in tetrahydrofuran (5 mL) was added to the reaction solution under ice bath. The reaction was stirred under ice bath for 1 hour. The reaction solution was quenched with saturated ammonium chloride. The aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined, washed with saturated sodium chloride, dried, and concentrated. The residue was chromatographed on a silica gel column (elution system B) to obtain (2,4-difluorophenyl)methanol-d2-ol 3B (0.4 g) with a yield of 94%.

[0315] Step 2

[0316] (2,4-Difluorophenyl)methanol-d2-ol 3B (400 mg, 2.74 mmol) and N,N-dimethylformamide (0.05 mL) were dissolved in dichloromethane (10 mL) and stirred. Sulfonyl chloride (749 mg, 6.30 mmol) was slowly added dropwise to the reaction solution under ice bath. The reaction was stirred at room temperature for 1 hour. The reaction solution was concentrated, and the residue was diluted with saturated sodium bicarbonate solution. The aqueous phase was extracted with dichloromethane (20 mL × 3), and the organic phases were combined, washed with saturated sodium chloride, dried, and concentrated to obtain 1-(chloromethyl-d2)-2,4-difluorobenzene intermediate 3 (0.42 g).

[0317] Intermediate 4

[0318] 2-(Chloromethyl-d2)-3,5-difluoropyridine

[0319]

[0320] first step

[0321] Lithium aluminum deuteride (388 mg, 9.24 mmol) was added in batches to a solution of methyl 3,5-difluoropyridine-2-carboxylate (1.00 g, 5.78 mmol) in tetrahydrofuran (20 mL) under an ice bath. The reaction was stirred at room temperature for 2 hours. The reaction was quenched with ice cubes and the reaction solution was filtered. The filtrate was concentrated and the residue was separated by silica gel column chromatography (eluent system B) to obtain the product (3,5-difluoropyridin-2-yl)methane-d2-ol 4A (345 mg) with a yield of 40.6%.

[0322] MS m / z(ESI):148.0[M+H] + .

[0323] Step 2

[0324] Thionyl chloride (839 mg, 7.06 mmol) was added dropwise to a solution of (3,5-difluoropyridin-2-yl)methane-d2-ol 4A (346 mg, 2.35 mmol) in dichloromethane (5 mL) and N,N-dimethylformamide (0.1 mL) under stirring. The reaction was stirred at room temperature for 3 hours. The reaction solution was diluted with ethyl acetate (160 mL), and the organic phase was washed with saturated sodium bicarbonate and water. The organic phase was concentrated, and the residue was separated by silica gel column chromatography (eluent system B) to give the product 2-(chloromethyl-d2)-3,5-difluoropyridine intermediate 4 (150 mg), yield: 38.5%.

[0325] MS m / z(ESI):166.0[M+H] + .

[0326] Intermediate 5

[0327] 2'-Bromo-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one

[0328]

[0329] first step

[0330] 1-(2-bromo-5-methyl-4-pyridyl)-3-chloro-4-hydroxy-6-methyl-pyridin-2-one 2C (550 g, 1.67 mmol), 2-(chloromethyl-d2)-3,5-difluoropyridine intermediate 4 (387 mg, 2.34 mmol), potassium carbonate (577 mg, 4.17 mmol) and 18-crown-6 ether (662 mg, 2.50 mmol) were dissolved in N,N-dimethylformamide (25 mL) and heated to 70 ° C and stirred for 3 hours. Ethyl acetate (50 mL) was added to the reaction solution, and the organic phase was washed with water and saturated sodium chloride, dried and concentrated. The residue was purified by silica gel column chromatography (elution system A) to give 2'-bromo-3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one intermediate 5 (540 mg) in a yield of 70.6%.

[0331] MS m / z(ESI):458.0[M+H] + .

[0332] Example 1

[0333] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-

[0334] [1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0335]

[0336] first step

[0337] Under ice bath, methyl 4-fluoro-1H-pyrazole-3-carboxylate (600 mg, 4.16 mmol) was dissolved in anhydrous tetrahydrofuran (20 mL), and a tetrahydrofuran solution of methyl magnesium bromide (3 M, 6.9 mL, 20.7 mmol) was added dropwise to the reaction solution under stirring, and the reaction was stirred at room temperature for 3 hours. The reaction solution was slowly poured into a saturated ammonium chloride solution (50 mL) under stirring, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with water (50 mL) and a saturated sodium chloride solution (50 mL) in sequence, dried, and concentrated to obtain 2-(4-fluoro-1H-pyrazol-3-yl)propane-2-ol 1a (600 mg), which was used directly in the next step without purification.

[0338] MS m / z(ESI):145.1[M+H] + .

[0339] Step 2

[0340] 1a (600 mg) was dissolved in acetonitrile (15 mL) and stirred. Concentrated sulfuric acid (5 mL) was added dropwise to the reaction solution at 0°C. The reaction was heated to room temperature and stirred for 3 hours. The reaction solution was slowly poured into a saturated sodium bicarbonate solution (100 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with water (50 mL) and a saturated sodium chloride solution (50 mL) in sequence, dried, and concentrated. The residue was purified by silica gel column chromatography (elution system B) to obtain N-(2-(4-fluoro-1H-pyrazol-3-yl)propane-2-yl)acetamide 1b (510 mg) with a yield of 86.8%.

[0341] MS m / z(ESI):186.1[M+H] + .

[0342] Step 3

[0343] Under nitrogen protection, intermediate 5 (80 mg, 0.174 mmol) was dissolved in 1,4-dioxane (2 mL), and then 1b (48 mg, 0.261 mmol), (trans)-dimethyl-1,2-cyclohexanediamine (12 mg, 0.087 mmol), cesium carbonate (170 mg, 0.523 mmol) and cuprous iodide (66 mg, 0.349 mmol) were added in sequence, and the reaction was heated to 100 ° C and stirred for 2 hours. The reaction solution was cooled to room temperature, poured into water (50 mL), and the aqueous phase was extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed successively with water (30 mL) and saturated sodium chloride solution (30 mL), dried, concentrated, and the residue was purified by reverse phase chromatography with eluent system C to give N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide 1 (75 mg), yield: 76.1%.

[0344] MS m / z(ESI):563.2[M+H] + .

[0345] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.56(d,1H),8.54(s,1H),8.11-8.06(m,2H),7.78 (s,1H),6.80(s,1H),2.01(s,3H),1.99(s,3H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0346] Decomposition of Example 1

[0347] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridin-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridin-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0348] fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0349]

[0350] Example 1 (40 mg, 0.071 mmol) was subjected to chiral separation (OD column) to give 1-1 (18.3 mg, RT = 3.020 min), yield: 45%; 1-2 (17.3 mg, RT = 3.960 min), yield: 43%.

[0351] Example 1-1: MS m / z (ESI): 563.2 [M+H] + ;

[0352] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.56(d,1H),8.54(s,1H),8.12-8.06(m,2H),7.78 (s,1H),6.80(s,1H),2.00(s,3H),1.99(s,3H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0353] Example 1-2: MS m / z (ESI): 563.2 [M+H] + ;

[0354] HPLC chiral separation conditions:

[0355]

[0356] HPLC chiral analysis method:

[0357]

[0358]

[0359] Example 2

[0360] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-

[0361] [1,4'-Bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0362]

[0363] first step

[0364] Intermediate 1 (400 mg, 3.17 mmol) and acetonitrile (781 mg, 19.02 mmol) were dissolved in acetic acid (1.52 g, 25.37 mmol) and stirred. Concentrated sulfuric acid (1.52 mL, 28.54 mmol) was added dropwise to the reaction solution at 0°C. The reaction temperature was raised to 60°C and stirred for 2 hours. Saturated sodium bicarbonate solution (40 mL) was added to the reaction solution at 0°C and stirred for 0.5 hours. The aqueous phase was extracted with ethyl acetate (25 mL × 3). The organic phases were combined, dried, concentrated, and prepared by reverse phase HPLC (ammonium bicarbonate system) to obtain N-(2-(1H-pyrazol-3-yl)propane-2-yl)acetamide 2a (53 mg) with a yield of 10%.

[0365] MS m / z(ESI):168.1[M+H] + ;

[0366] Step 2

[0367] Under nitrogen protection, N-(2-(1H-pyrazol-3-yl)propan-2-yl)acetamide 2a (17.9 mg, 0.107 mmol), intermediate 2 (35 mg, 0.077 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (2 mg, 0.015 mmol), cuprous iodide (3 mg, 0.015 mmol) and cesium carbonate (50 mg, 0.153 mmol) were dissolved in 1,4-dioxane (1.5 mL), and the reaction was heated to 100 ° C and stirred for 16 hours. Saturated ammonium chloride solution (30 mL) was added to the reaction solution, the aqueous phase was extracted with ethyl acetate (15 mL×2), the organic phases were combined, dried and concentrated, and the residue was separated by silica gel column chromatography (elution system B) to give N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide 2 (28.6 mg), yield: 68.8%.

[0368] MS m / z(ESI):543.1[M+H] + ;

[0369] Decomposition of Example 2

[0370] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0371] 2-YL)PROPAN-2-YL)ACETAMIDE

[0372]

[0373] Example 2 (45 mg, 0.083 mmol) was subjected to chiral separation (OD column) to give the title product 2-1 (14.9 mg, RT = 3.14 min), yield: 33.11%; 2-2 (13.9 mg, RT = 3.62 min), yield: 30.89%.

[0374] Example 2-1, MS m / z (ESI): 543.1 [M+H] + ;

[0375] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.54(s,1H),8.48(d,1H),8.14-8.04(m,1H),7.93(s,1H),7.77 (s,1H),6.80(s,1H),6.42(d,1H),5.48(d,2H),2.01(s,6H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0376] Example 2-2, MS m / z (ESI): 543.1 [M+H] + ;

[0377] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.54(s,1H),8.48(d,1H),8.14-8.04(m,1H),7.93(s,1H),7.77 (s,1H),6.80(s,1H),6.42(d,1H),5.48(d,2H),2.01(s,6H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0378] HPLC chiral separation conditions:

[0379]

[0380] HPLC chiral analysis method:

[0381]

[0382] Example 3

[0383] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0384]

[0385] Referring to the synthesis method of Example 2, the target product N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide 3 was synthesized.

[0386] MS m / z(ESI):545.2[M+H] + ;

[0387] Decomposition of Example 3

[0388] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-1H-

[0389] Pyrazol-3-yl)propan-2-yl)acetamide

[0390]

[0391] Example 3 (75 mg, 0.138 mmol) was subjected to chiral separation (OD column) to give the title product 3-1 (24.3 mg, RT = 3.123 min), yield: 32.4%; 3-2 (24.6 mg, RT = 3.627 min), yield: 32.8%.

[0392] Example 3-1, MS m / z (ESI): 545.2 [M+H] + ;

[0393] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.54(s,1H),8.48(d,1H),8.13-8.06(m,,1H),7.93(s,1H) ),7.77(s,1H),6.80(d,1H),6.42(d,1H),2.01(s,6H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0394] Example 3-2, MS m / z (ESI): 545.2 [M+H] + ;

[0395] 1 H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.54(s,1H),8.48(d,1H),8.13-8.06(m,,1H),7.93(s,1H) ),7.77(s,1H),6.80(d,1H),6.42(d,1H),2.01(s,6H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0396] The HPLC chiral separation conditions and HPLC chiral analysis method are shown in Example 2.

[0397] Example 4

[0398] N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-

[0399] [bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0400]

[0401] first step

[0402] Under nitrogen protection, 2C (400 mg, 1.21 mmol), potassium carbonate (503 mg, 3.64 mmol) and 18-crown-6 (481 mg, 1.82 mmol) were dissolved in anhydrous N,N-dimethylformamide (6 mL), heated to 75 ° C and stirred, and intermediate 3 (300 mg, 1.82 mmol) was added dropwise to the reaction solution. The reaction was stirred at 75 ° C for 1 hour. Water (50 mL) was added to the reaction solution, and the aqueous phase was extracted with ethyl acetate (50 mL × 2). The organic phases were combined, washed successively with water (50 mL) and saturated sodium chloride solution (50 mL), dried, and the filtrate was concentrated. The residue was separated by silica gel column chromatography (elution system B) to give 2'-bromo-3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 4a (480 mg), yield: 86.4%.

[0403] MS m / z(ESI):457.0[M+H] + .

[0404] Step 2

[0405] Under nitrogen protection, 2'-bromo-3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2H-[1,4'-bipyridyl]-2-one 4a (100 mg, 0.218 mmol), 2a (73 mg, 0.437 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (31 mg, 0.218 mmol), cuprous iodide (42 mg, 0.218 mmol) and cesium carbonate (142 mg, 0.437 mmol) were dissolved in 1,4-dioxane (2 mL), and the reaction was heated to 100 ° C and stirred for 2 hours. Saturated sodium bicarbonate solution (30 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL×2). The organic phases were combined, dried, and concentrated. The residue was separated by silica gel column chromatography to give N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide 4 (75 mg), yield: 63.10%.

[0406] MS m / z(ESI):544.2[M+H] + .

[0407] Decomposition of Example 4

[0408] (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0409]

[0410] Example 4 (75 mg, 0.138 mmol) was subjected to chiral separation (AD column) to give the title product 4-1 (27.2 mg, RT = 4.827 min), yield: 36.27%; 4-2 (27.9 mg, RT = 5.697 min), yield: 37.2%.

[0411] Example 4-1, MS m / z (ESI): 544.2 [M+H] + ;

[0412] 1 H NMR(400MHz,DMSO-d6)δ8.55(s,1H),8.49(d,1H),7.96(s,1H),7.78(s,1H),7.76-7.68(m,1H),7.40-7.35(m,1H ),7.25-7.19(m,1H),6.83(d,1H),6.43(d,1H),2.02(s,3H),2.01(s,3H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0413] Example 4-2, MS m / z (ESI): 544.2 [M+H] + ;

[0414] 1 H NMR(400MHz,DMSO-d6)δ8.55(s,1H),8.49(d,1H),7.96(s,1H),7.78(s,1H),7.76-7.68(m,1H),7.40-7.35(m,1H ),7.25-7.19(m,1H),6.83(d,1H),6.43(d,1H),2.02(s,3H),2.01(s,3H),1.82(s,3H),1.61(s,3H),1.57(s,3H).

[0415] The HPLC chiral separation conditions and HPLC chiral analysis method are shown in Example 2.

[0416] Example 5

[0417] N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-

[0418] [bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0419]

[0420] Referring to the synthesis method of Example 4, the target product N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridine]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide 5 was synthesized.

[0421] MS m / z(ESI):562.1[M+H] + .

[0422] Decomposition of Example 5

[0423] (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0424] Pyrazol-3-yl)propan-2-yl)acetamide

[0425]

[0426] Example 5 (60 mg, 0.107 mmol) was subjected to chiral separation (OD column) to give 5-1 (19.4 mg, RT = 4.397 min), yield: 32%; 5-2 (9.7 mg, RT = 5.757 min), yield: 16%.

[0427] Example 5-1: MS m / z (ESI): 562.2 [M+H] + ;

[0428] 1H NMR(400MHz,DMSO-d6)δ8.57(d,1H),8.55(s,1H),8.11(s,1H),7.79(s,1H),7.74-7.68(m,1H),7.42-7.39 (m,1H),7.24-7.19(m,1H),6.83(s,1H),2.02(s,3H),2.01(s,3H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0429] Example 5-2: MS m / z (ESI): 562.2 [M+H] + ;

[0430] 1 H NMR(400MHz,DMSO-d6)δ8.57(d,1H),8.55(s,1H),8.11(s,1H),7.79(s,1H),7.73-7.68(m,1H),7.41-7.39 (m,1H),7.25-7.18(m,1H),6.83(s,1H),2.02(s,3H),2.01(s,3H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0431] The HPLC chiral separation conditions and HPLC chiral analysis method are shown in Example 1.

[0432] Example 6

[0433] N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0434]

[0435] Referring to the synthesis method of Example 4, the target product N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridine]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide 6 was synthesized.

[0436] MS m / z(ESI):580.2[M+H] + .

[0437] Decomposition of Example 6

[0438] (S)-N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0439]

[0440] Example 6 (60 mg, 0.107 mmol) was subjected to chiral separation (OD column) to give 6-1 (19.4 mg, RT = 4.397 min), yield: 32%; 6-2 (9.7 mg, RT = 5.757 min), yield: 16%.

[0441] Example 6-1: MS m / z (ESI): 580.2 [M+H] + ;

[0442] 1 H NMR(400MHz,DMSO-d6)δ8.57(d,1H),8.54(s,1H),8.10(s,1H),7.80(s,1H),7.3 9-7.34(m,2H),6.84(s,1H),2.01(s,6H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0443] Example 6-2: MS m / z (ESI): 580.2 [M+H] + ;

[0444] 1 H NMR(400MHz,DMSO-d6)δ8.57(d,1H),8.54(s,1H),8.10(s,1H),7.80(s,1H),7.3 9-7.34(m,2H),6.84(s,1H),2.01(s,6H),1.80(s,3H),1.60(s,3H),1.57(s,3H).

[0445] The HPLC chiral separation conditions were as shown in Example 1, wherein the mobile phase was changed to Hexane:EtOH (+0.1% DEA) = 85:15; the HPLC chiral analysis method was as shown in Example 1, wherein the mobile phase was changed to Hexane:EtOH (+0.1% DEA) = 85:15.

[0446] Example 7

[0447] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-

[0448] bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide

[0449]

[0450] first step

[0451] Sulfuric acid (3.40 g, 34.69 mmol) was added dropwise to a solution of 1a (500 mg, 3.47 mmol) in 3-bromopropionitrile (4.65 g, 34.69 mmol) under ice bath and stirred. The reaction solution was heated to 30°C and stirred for 16 hours. The reaction solution was concentrated and the residue was separated using Prep-HPLC column method (formic acid system) to obtain the title product 3-bromo-N-(2-(4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide 7b (520 mg) with a yield of 53.9%.

[0452] MS m / z(ESI):278.0[M+H] + .

[0453] Step 2

[0454] 3-Bromo-N-(2-(4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide 7b (500 mg, 1.80 mmol) and zinc powder (1.17 g, 17.98 mmol) were dissolved in acetic acid (10 mL) and tetrahydrofuran (5 mL) solution, and the reaction was stirred at 25°C for 16 hours. The reaction solution was filtered, the filtrate was concentrated, and the residue was separated by Prep-HPLC column method (formic acid system) to obtain the title product N-(2-(4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide 7c (260 mg), with a yield of 72.6%.

[0455] MS m / z(ESI):200.1[M+H] + .

[0456] Step 3

[0457] N-(2-(4-Fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide 7c (65 mg, 0.33 mmol), intermediate 5 (100 mg, 0.22 mmol), (1S,2S)-N1,N2-dimethylcyclohexane-1,2-diamine (12 mg, 0.09 mmol), cesium carbonate (106 mg 0.33 mmol), and cuprous iodide (17 mg, 0.09 mmol) were dissolved in dioxane (2 mL), and the reaction was heated to 100 °C and stirred for 3 hours. The reaction solution was filtered, the filtrate was concentrated, and the residue was separated using normal phase silica gel column chromatography (eluent system A) to give the title product N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide 7 (80 mg), yield: 63.6%.

[0458] MS m / z(ESI):577.2[M+H] + .

[0459] Decomposition of Example 7

[0460] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridin-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridin-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide

[0461] fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide

[0462]

[0463] Example 7 (80 mg, 0.138 mmol) was subjected to chiral separation (OD column) to give 7-1 (31 mg, RT = 2.840 min), yield: 38%; 7-2 (27 mg, RT = 3.380 min), yield: 33%.

[0464] Example 7-1: MS m / z (ESI): 577.2 [M+H] + ;

[0465] 1H NMR (400MHz, DMSO-d6) δ8.59(m,3H),8.11(m,1H),8.02(s,1H),7.79(s,1H),6.81(d,1H),2.09(q,2H),2.01(d,6H),1.59(d,6H),0.95(t,3H).

[0466] Example 7-2: MS m / z (ESI): 577.2 [M+H] + ;

[0467] 1 H NMR (400MHz, DMSO-d6) δ8.59(m,3H),8.11(m,1H),8.02(s,1H),7.79(s,1H),6.81(d,1H),2.09(q,2H),2.01(d,6H),1.59(d,6H),0.95(t,3H).

[0468] HPLC chiral separation conditions:

[0469]

[0470] HPLC chiral analysis conditions:

[0471] instrument HPLC (Waters) Column Type DAICEL CHIRALCEL OD-H 4.6mm ID×150mmL 5.0μm Column pressure 45psi Mobile phase Hexane:EtOH=70:30(V:V) Flow rate 1mL / min Detection wavelength UV 214nm Column temperature 35℃

[0472] Example 8

[0473] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-

[0474] [1,4'-Bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3

[0475]

[0476] Referring to the synthesis method of Example 1, the target product N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3 8 was synthesized.

[0477] MS m / z(ESI):566.2[M+H] + .

[0478] Decomposition of Example 8

[0479] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3 and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3

[0480]

[0481] Example 8 (60 mg, 0.106 mmol) was subjected to chiral separation (OD column) to give 8-1 (23 mg, RT = 3.003 min), yield: 38%; 8-2 (27 mg, RT = 3.943 min), yield: 33%.

[0482] Example 8-1: MS m / z (ESI): 566.2 [M+H] + ;

[0483] 1 H NMR (400MHz, CDCl3) δ8.36(m,3H),7.67(s,1H),7.33(m,1H),6.41(d,1H),6.34(s,1H),2.06(d,6H),1.76(d,6H).

[0484] Example 8-2: MS m / z (ESI): 566.2 [M+H] + ;

[0485] 1 H NMR (400MHz, CDCl3) δ8.36(m,3H),7.67(s,1H),7.33(m,1H),6.41(d,1H),6.34(s,1H),2.06(d,6H),1.76(d,6H).

[0486] The HPLC chiral separation conditions and HPLC chiral analysis conditions are shown in Example 7.

[0487] Example 9

[0488] N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl

[0489] -2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0490]

[0491] Referring to the synthesis method of Example 1, the target product N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide 9 was synthesized.

[0492] MS m / z(ESI):579.1[M+H] + .

[0493] Decomposition of Example 9

[0494] (S)-N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-

[0495] 2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0496]

[0497] Example 9 (70 mg, 0.107 mmol) was subjected to chiral separation (OJ column) to give 9-1 (34.3 mg, RT = 3.080 min), yield: 48%; 9-2 (30.2 mg, RT = 5.023 min), yield: 42%.

[0498] Example 9-1: MS m / z (ESI): 579.1 [M+H] + ;

[0499] 1 H NMR(400MHz,DMSO-d6)δ8.67(s,1H),8.61(d,1H),8.56(s,1H),8.14(s,1H),8.14-8.07(m,1H ),7.81(s,1H),6.81(s,1H),2.01(s,3H),2.00(s,3H),1.81(s,3H),1.60(s,3H),1.56(s,3H).

[0500] Example 9-2: MS m / z (ESI): 579.1 [M+H] + ;

[0501] 1 H NMR(400MHz,DMSO-d6)δ8.67(s,1H),8.61(d,1H),8.56(s,1H),8.14(s,1H),8.12-8.09(m,1H ),7.81(s,1H),6.81(s,1H),2.01(s,3H),2.00(s,3H),1.81(s,3H),1.60(s,3H),1.56(s,3H).

[0502] HPLC chiral separation conditions:

[0503]

[0504] HPLC chiral analysis method:

[0505]

[0506] Example 10

[0507] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridinyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0508]

[0509] Referring to the synthesis method of Example 1, the target product N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide 10 was synthesized.

[0510] MS m / z(ESI):559.2[M+H] + .

[0511] 1H NMR(400MHz,DMSO-d6)δ8.60(d,1H),8.49(s,1H),8.28(d,1H),8.09(ddd,1H),8.04(s,1 H),7.70(s,1H),6.80(d,1H),2.06(d,3H),2.02–1.97(m,6H),1.58(s,3H),1.53(s,3H).

[0512] Decomposition of Example 10

[0513] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridinyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridinyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide

[0514]

[0515] Example 10 (120 mg, 0.215 mmol) was subjected to chiral separation (OD column) to give 10-1 (53 mg, RT = 3.000 min), yield: 44%; 10-2 (56 mg, RT = 3.577 min), yield: 46%.

[0516] Example 10-1: MS m / z (ESI): 559.2 [M+H] + ;

[0517] Example 10-2: MS m / z (ESI): 559.2 [M+H] + ;

[0518] The HPLC chiral separation conditions were as shown in Example 1, wherein the mobile phase was changed to Hexane:EtOH:MeOH:DEA=70:15:15:0.1%; the HPLC chiral analysis method was as shown in Example 1, wherein the mobile phase was changed to Hexane:EtOH:MeOH:DEA=70:15:15:0.1%.

[0519] Embodiment 11

[0520] N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-

[0521] [1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide

[0522]

[0523] Referring to the synthesis method of Example 1, the target product N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide 11 was synthesized.

[0524] MS m / z(ESI):589.2[M+H] + .

[0525] Decomposition of Example 11

[0526] (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide and (R)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridinyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide

[0527]

[0528] Example 11 (60 mg, 0.102 mmol) was subjected to chiral separation (OD column) to give 11-1 (22 mg, RT = 2.923 min), yield: 36.7%; 11-2 (23 mg, RT = 4.257 min), yield: 38.3%.

[0529] Example 11-1: MS m / z (ESI): 589.2 [M+H] + ;

[0530] 1 H NMR (400MHz, DMSO-d6) δ8.59(m,3H),8.30(s,1H),8.11(m,1H),7.79(s,1H),6.81(s,1H),2.00(d,6H),1.61(d,7H),0.58(m,4H).

[0531] Example 11-2: MS m / z (ESI): 589.2 [M+H]+ ;

[0532] 1 H NMR (400MHz, DMSO-d6) δ8.59(m,3H),8.30(s,1H),8.11(m,1H),7.79(s,1H),6.81(s,1H),2.00(d,6H),1.61(d,7H),0.58(m,4H).

[0533] The HPLC chiral separation conditions and HPLC chiral analysis conditions are shown in Example 7.

[0534] Example Test Evaluation

[0535] The present invention is further described and explained below in conjunction with test examples, but these embodiments are not intended to limit the scope of the present invention.

[0536] 1. Experimental instruments and reagents

[0537] 1. Instrument

[0538] EnVision (PerkinElmer, ENVISION 2015 or 2105-0020)

[0539] 2. Reagents

[0540] Active p38 alpha Protein (R&D systems, 5477-KS-010), Inactive MK2 (Thermo, PV3316), ULight-CREBtide (Perkinelmer, TRF0107), ATP Solution (Invitrogen, PV3227), DMSO (Sigma, D2650), HEPES (1M) (Gibco, 15630-080), MgCl2 (1M) (Invitrogen, AM9530G), Tween 20 (Sigma, P9416), EGTA (Alfa Aesar, J60767), DTT (1M) (Sigma, 43816), Europium-anti-phospho-CREB (PerkinElmer, TRF0200), EDTA (0.5M) (Invitrogen, AM9260G), 10X LANCE detection buffer (PerkinElmer, CR97-100), 384-well plate (Perkinelmer, 6007299), 96V bottom plate (PS) (Axygen, WIPP02280), U937 (ATCC, CRL-1593.2), Phospho-HSP27 (S78 / S82) DuoSet IC ELISA (R&D systems, DYC2314-5), DuoSet ELISA Ancillary Reagent Kit 2 (R&D systems, DY008), Phorbol 12-myristate 13-acetate (PMA) (Abcam, Ab120297), Lipopolysaccharides from Escherichia coli O111:B4 (Sigma-Aldrich, L4391), Human TNF-alpha DuoSet ELISA (R&D systems, DY210).

[0541] 2. Test Method

[0542] Test Example 1. Determination of the inhibitory effect of the compounds of the present invention on p38α / MK2 enzyme activity 1. Experimental purpose: The purpose of this test example is to measure the inhibitory ability of the compounds on p38α / MK2 enzyme activity. 2. Experimental method:

[0543] p38 / MK2 enzyme activity assay method: In 1× enzyme reaction buffer (50mM HEPES pH 7.5, 1mM EGTA, 10mMMgCl2, 2mM DTT, 0.01% Tween-20), incubate 0.06nM p38α (Active p38 alpha Protein), 1.2nM MK2 (Inactive MK2) and gradient dilutions of compounds for 1 hour. Add 50nM ULight-CREBtide and 25μM ATP (ATP Solution) and react at room temperature for 1.5 hours. Add an equal volume of detection and termination mixture (1×LANCE detection buffer containing 20mMEDTA and 2nM Europium-anti-phospho-CREB), mix and centrifuge, apply sealing film, and react at room temperature for 1 hour. Read the plate using the TR-FRET program of the EnVision instrument and detect the fluorescence value of each well at 665nm / 615nm.

[0544] p38 enzyme activity assay method: Incubate 3nM p38α (Active p38 alpha Protein) and gradient dilutions of compounds for 1 hour in 1× enzyme reaction buffer (50mM HEPES pH 7.5, 1mM EGTA, 10mMMgCl2, 2mM DTT, 0.01% Tween-20). Add 50nM ULight-MBP and 50μM ATP (ATP 10mM Solution) and react at room temperature for 1.5 hours. Add an equal volume of detection and termination mixture (1×LANCE detection buffer containing 20mM EDTA and 2nM Europium-anti-P-MBP), mix and centrifuge, apply sealing film, and react at room temperature for 1 hour. Read the plate using the TR-FRET program of the EnVision instrument and detect the fluorescence value of each well at 665nm / 615nm.

[0545] 3. Experimental data processing method:

[0546] The raw data (665nm / 615nm reading ratio) was calculated according to the following formula to obtain the inhibition rate: Inhibition rate % = [(average value of positive control wells - sample wells) / (average value of positive control wells - average value of negative control wells)] × 100, where the positive control wells are reaction wells without compound enzymes, and the negative control wells are reaction wells without enzymes.

[0547] Use log(inhibitor) vs.response--Variable slope(four parameters) in GraphPad Prism 6 to fit the compound concentration and the corresponding inhibition rate, fit the curve and obtain the compound IC 50 value.

[0548] The fitting calculation equation is Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0549] 4. Experimental results:

[0550] Table 6

[0551] Example <![CDATA[p38 / MK2 IC 50 (nM)]]> <![CDATA[p38 IC 50 (nM)]]> p38 / p38 / MK2 1-1 0.5 83.7 167 2-1 1.1 / / 3-1 1.3 113.7 87 4-2 0.1 / / 5-1 0.2 / / 6-1 0.2 / / 7-1 0.9 97.9 109 8-1 0.7 73.0 104 11-1 0.6 103.9 173

[0552] 5. Experimental conclusion: The above data show that the compounds of the present invention have strong inhibitory ability on p38α / MK2 enzyme activity and good selectivity.

[0553] Test Example 2: Determination of the inhibitory effect of the compounds of the present invention on p-Hsp27 in U937 cells 1. Experimental purpose: The purpose of this test example is to test the inhibitory ability of the compounds of the present invention on p-Hsp27 in U937 cells. 2. Experimental method:

[0554] U937 cells were treated with 20ng / ml PMA for differentiation overnight, and then fresh culture medium was replaced. U937 cells were rested overnight. U937 cells were digested and counted after differentiation, and 25,000 cells were seeded in 96-well plates per well until the cells adhered to the wall. Compound concentration gradients were prepared, and cells were pretreated with compounds for 1 hour (the highest final concentration of the compound was 1μM, and the concentration gradient was diluted 3 times), and 10ng / ml LPS (Lipopolysaccharides from Escherichia coli O111:B4) was added for stimulation for 0.5 hours. After washing the cells with PBS, the cells were lysed with lysis buffer. The p-Hsp27 in the cell lysate was detected using the Phospho-HSP27 (S78 / S82) DuoSet IC ELISA kit. The specific steps refer to the standard steps provided by the kit manufacturer.

[0555] 3. Experimental data processing method:

[0556] Calculate the p-Hsp27 inhibition rate: inhibition rate percentage = [(average p-Hsp27 in positive control wells - average p-Hsp27 in sample wells) / (average p-Hsp27 in positive control wells - average p-Hsp27 in negative control wells)] × 100%, where the positive control wells are wells without compound (DMSO-treated wells) and the negative control wells are wells without LPS stimulation.

[0557] Dose response curve fitting: Use log(inhibitor) vs.response--Variable slope(four parameters) in GraphPad Prism 6 to fit the compound concentration and the corresponding inhibition rate, fit the curve and obtain the compound IC 50 value.

[0558] The fitting calculation equation is Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0559] 4. Experimental results:

[0560] Table 7

[0561]

[0562] 5. Experimental conclusion: The above data show that the compounds of the present invention have a strong inhibitory ability on U937 cell p-Hsp27.

[0563] Test Example 3: Determination of the inhibitory effect of the compounds of the present invention on the expression level of TNF-α in U937 cells

[0564] 1. Experimental purpose: The purpose of this test example is to measure the inhibitory ability of the compounds of the present invention on the expression level of TNF-α in U937 cells.

[0565] 2. Experimental methods:

[0566] U937 cells were treated with 20ng / ml PMA for differentiation overnight, and then fresh culture medium was replaced. U937 cells were rested overnight. U937 cells were digested and counted after differentiation, and 50,000 cells were seeded in 96-well plates per well until the cells adhered to the wall. Compound concentration gradients were prepared, and cells were pretreated with compounds for 1 hour (the highest final concentration of the compound was 1μM, and the concentration gradient was diluted 3 times). After adding 10ng / ml LPS (Lipopolysaccharides from Escherichia coli O111:B4) for stimulation for 4 hours, the culture supernatant was collected by centrifugation for subsequent ELISA detection of TNF-α concentration in the supernatant. TNF-α concentration detection was performed according to the standard steps provided by the ELISA kit manufacturer.

[0567] 3. Experimental data processing method:

[0568] The standard curve of TNF-α standard was drawn by linear regression method, and the TNF-α concentration of the sample was converted according to the standard curve.

[0569] Calculate the inhibition rate: inhibition rate percentage = [(average TNF-α concentration in positive control wells - average TNF-α concentration in sample wells) / (average TNF-α concentration in positive control wells - average TNF-α concentration in negative control wells)] × 100%, where the positive control wells are compound-free wells (DMSO-treated wells) and the negative control wells are culture medium wells.

[0570] Dose response curve fitting: Use log(inhibitor) vs.response–Variable slope(four parameters) in GraphPad Prism 6 to fit the compound concentration and the corresponding inhibition rate, fit the curve and obtain the compound IC 50 The fitting calculation equation is Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)).

[0571] 4. Experimental results:

[0572] Table 8

[0573]

[0574] 5. Experimental conclusion: The above data show that the compounds of the present invention have a strong inhibitory ability on the expression level of TNF-α in U937 cells.

[0575] Test Example 4: Investigation of in vitro metabolic stability of the compounds of the present invention in human liver microsomes

[0576] 1. Experimental purpose: The purpose of this study is to evaluate the metabolic stability of the compound in phase I and part of phase II in human liver microsomes.

[0577] 2. Experimental Plan

[0578] 2.1 Trial Drugs:

[0579] The compound of the present invention is self-made.

[0580] Alamethicin (Abcam), 7-Hydroxycoumarin (Sigma), liver microsomes (XenoTech, Shanghai Quanyang Biotechnology Co., Ltd.), phosphate buffer (Gibco, Lot#SLBS7904 and Lot#SLBR3106V, pH 7.4), NADPH (reduced nicotinamide adenine dinucleotide phosphate, Shanghai BiDe Pharmaceutical Technology Co., Ltd.), UDPGA (Sigma), Alamethicin (Abcam), methanol (Merck), acetonitrile (Merck).

[0581] 2.2 Drug preparation: The compound of the present invention is prepared into a 10 mM stock solution with DMSO (or other suitable solution) and stored in a -20°C refrigerator until use.

[0582] 2.3 Experimental procedures

[0583] 1) Prepare buffer:

[0584] Take 4.01 mL of 1 M K2HPO4·3H2O (AR grade) and 0.99 mL of 1 M KH2PO4 (AR grade), dissolve them in ultrapure water and make up to 50 mL to prepare a phosphate buffer with a final concentration of 100 mM.

[0585] 2) Prepare compound working solution

[0586] Preparation of compound working solution: Add 2 μL of compound stock solution to 998 μL of phosphate buffer, with a final concentration of 20 μM. Depending on the properties of the compound, the configuration ratio can be adjusted appropriately to adjust the final concentration.

[0587] 3) Preparation of liver microsome working solution

[0588] 156.3 μL of 20 mg / mL microsomes were diluted to 5 mL with 100 mM phosphate buffer and mixed to a final concentration of 0.625 mg / mL.

[0589] 4) Prepare NADPH and UDPGA

[0590] Weigh 33.3 mg of NADPH and 25.8 mg of UDPGA, add 2 mL of 100 mM phosphate buffer, and the final concentration of both is 20 mM.

[0591] 5) Prepare the punch (Alamethicin)

[0592] Weigh 1 mg of Alamethicin and add 200 μL of methanol to prepare a 5 mg / mL solution. Then take out 10 μL of the solution and add it to 990 μL of phosphate buffer (pH 7.4) to a final concentration of 50 μg / mL.

[0593] 6) Prepare the reaction termination solution

[0594] Dilute the internal standard with acetonitrile (or other suitable solution) as the stop solution and store it in a refrigerator at 2-8°C.

[0595] 7) Incubation process

[0596] 400 μL of prepared liver microsomes, 25 μL of compound working solution (10 μM) and 25 μL of Lamethicin (50 μg / mL) were added to a 96-well plate and pre-incubated at 37°C for 10 min. Then 50 μL of prepared NADPH / UDPGA was added to start the reaction and incubated at 37°C. The total volume of the reaction system was 500 μL, and the final contents of each component were as follows:

[0597] Element content Liver microsomes 0.5mg / mL Compound 0.5μM NADPH 1mM UDPGA 1mM Alamethicin 2.5 μg / mL

[0598] 50 μL was taken out at 0, 5, 15, 30, 60 and 120 min, and 200 μL of cold stop solution containing internal standard was added to terminate the sample reaction. The sample was centrifuged at 3500 rpm for 10 min, and the supernatant was taken for LC-MS / MS analysis.

[0599] 2.4 Chromatographic analysis

[0600] 1) Chromatographic conditions

[0601] Instrument: Shimadzu LC-20AD;

[0602] Column: Phenomenex C18 (50*4.6mm, 5μm particle size);

[0603] Mobile phase: A: 0.1% formic acid aqueous solution, B: acetonitrile;

[0604] Washing gradient: 0.2-1.6 min 5% A to 95% A, 3.0-3.1 min 95% A to 5% A;

[0605] Flow rate: 1.0ml / min;

[0606] Running time: 4.0min;

[0607] Injection volume: 5 μL.

[0608] 2) Mass spectrometry conditions

[0609] Instrument: API4000 liquid chromatography-mass spectrometer, AB Sciex;

[0610] Ion source: electrospray ionization source (ESI);

[0611] Drying gas: N2, temperature 500℃;

[0612] Electrospray voltage: 5000 V;

[0613] Detection method: positive ion detection;

[0614] Scanning mode: reaction monitoring (MRM) mode;

[0615] Scan time: 0.8401s.

[0616] 3. Experimental results and data processing

[0617] T 1 / 2 =0.693 / Ke, where Ke represents the elimination rate constant.

[0618] Table 9

[0619]

[0620] 4. Experimental conclusion: The above data show that the compounds of the present invention exhibit good metabolic stability in human liver microsomes.

[0621] Test Example 5: Pharmacokinetics in SD rats

[0622] 1. Experimental purpose: SD rats were used as test animals to study the pharmacokinetic behavior of the compound of the present invention in rats (plasma) at a dose of 5 mg / kg orally and 1 mg / kg intravenously.

[0623] 2. Experimental Plan

[0624] 2.1 Test drugs: Compounds of the present invention, homemade.

[0625] 2.2 Experimental animals: 3 SD rats per group, male, Shanghai Jiesijie Experimental Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006N0.311620400001794).

[0626] 2.3 Drug preparation:

[0627] PO, 10% solutol HS15, dissolve by ultrasound, prepare as a clear solution or homogeneous suspension.

[0628] IV, 5% DMSO + 10% Solutol HS15 + 85% PBS, sonicated to prepare a clear solution.

[0629] 2.4 Dosage regimen: 3 male SD rats per group; after fasting overnight, the rats were administered orally at a dose of 5 mg / kg in a volume of 10 mL / kg; and iv at a dose of 1 mg / kg in a volume of 5 mL / kg.

[0630] 2.5 Sample collection: After oral administration to rats, at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours; after intravenous administration to rats, at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours; 0.2 mL of blood was collected from the jugular vein, placed in an EDTA-K2 test tube, centrifuged at 6000 rpm at 4°C for 6 min to separate plasma, and stored at -80°C.

[0631] 2.6 Sample processing:

[0632] 1) Add 100uL acetonitrile to 20uL of plasma sample for precipitation, mix and centrifuge at 3500×g for 5 to 20 minutes.

[0633] 2) The supernatant solution after treatment is subjected to LC / MS / MS analysis to determine the concentration of the test compound. The LC / MS / MS analysis instrument is AB Sciex API 4000Qtrap.

[0634] 2.7 Liquid phase analysis:

[0635] Liquid phase conditions: Shimadzu LC-20AD pump

[0636] ●Chromatographic column: Waters Xbridge C18 5μm, 4.6X 50mm Mobile phase: Liquid A is 0.1% formic acid aqueous solution, Liquid B is methanol

[0637] Flow rate: 1.0mL / min

[0638] Elution time: 0-4.0 minutes, eluent is as follows:

[0639]

[0640] 3. Test results and data processing:

[0641] The main pharmacokinetic parameters were calculated using WinNonlin 6.1.

[0642] Table 10

[0643]

[0644] 4. Test conclusion:

[0645] The results of the pharmacokinetic assay in SD rats showed that the compound of the present invention exhibited a good PK advantage. The pharmacokinetics of the compound of the present invention were also determined in mice and beagle dogs, and the results showed that there was little difference in the PK species of the compound of the present invention.

[0646] Test Example 6: Pharmacokinetics in SD rats

[0647] 1. Experimental purpose: SD rats were used as test animals to study the pharmacokinetic behavior of the compound of the present invention in rats (plasma and brain) at a dose of 5 mg / kg after oral administration.

[0648] 2. Experimental Plan

[0649] 2.1 Test drugs: Compounds of the present invention, homemade.

[0650] 2.2 Experimental animals: 12 male SD rats per group, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0651] 2.3 Drug preparation: PO, 10% solutol HS15, dissolve by ultrasound, and prepare into a clear solution or homogeneous suspension.

[0652] 2.4 Dosage regimen: 12 male SD rats per group; after fasting overnight, the dose was 5 mg / kg, and the administration volume was 10 mL / kg.

[0653] 2.5 Sample collection: After oral administration of the rats, plasma and brain tissue were collected at 0.5, 1, and 2 hours; 0.25 mL of blood was collected from the jugular vein and anticoagulated with sodium heparin. The blood samples were placed on ice after collection and centrifuged within 1 hour to separate the plasma (centrifugation conditions: 6000g, 3 minutes, 2-8°C). Plasma samples were stored in a -80°C refrigerator before analysis.

[0654] 2.6 Sample processing:

[0655] 1) 50uL of plasma sample was added with 500uL of 50% methanol-acetonitrile solution containing internal standard (100ng / mL), mixed, centrifuged at 4000rpm and 4°C for 10 minutes, and analyzed by LC-MS / MS.

[0656] 2) After all samples are completely thawed, weigh a certain amount of brain tissue, add 4 times the volume of 50% methanol / water, and homogenize. Transfer 50uL of the homogenized brain tissue sample to a 96-well plate, and add 500uL of 50% methanol acetonitrile solution containing internal standard (100ng / mL). Vortex the sample for 5 minutes, centrifuge at 4000rpm and 4℃ for 10 minutes, and analyze by LC-MS / MS.

[0657] 3) Pipette 80uL of supernatant into 80uL of water, mix well and perform LC / MS / MS analysis on the concentration of the compound to be tested. LC / MS / MS analysis instrument: AB Sciex Triple Quad 5500+.

[0658] 2.7 Liquid phase analysis:

[0659] ·Chromatographic column: Synergi 4μm Fusion-RP 80A LC Column 50*2mm Mobile phase: Liquid A is 0.1% formic acid aqueous solution, Liquid B is acetonitrile

[0660] Flow rate: 0.8mL / min

[0661] Elution time: 0-2.2 minutes, eluent is as follows:

[0662]

[0663]

[0664] 3. Experimental results and data processing:

[0665] The main pharmacokinetic parameters were calculated using WinNonlin 6.1.

[0666] Table 11

[0667]

[0668] 5. Experimental Conclusion

[0669] The results of pharmacokinetic measurements in SD rats showed that the compound of the present invention had an extremely low drug concentration in the brain. The low brain-blood ratio indicated that the compound of the present invention had a poor ability to enter the brain, and the side effects such as headaches it caused were likely to be lower.

[0670] Test Example 7: PD test in Wistar rat acute inflammation model

[0671] 1. Experimental purpose: Using 6-8 week old Wistar rats as test animals, the pharmacodynamic behavior of the compound of the present invention in rats (serum) after oral administration at different doses and time points was studied.

[0672] 2. Experimental Plan

[0673] 2.1 Test sample: Compounds of the present invention, homemade. Wistar rats, Vital River Company.

[0674] 2.2 Drug preparation: Weigh an appropriate amount of compound, add the corresponding solvent 10% Solutol HS15, and obtain a suspension with a final concentration of 0.2 mg / mL by ultrasound, vortex and stirring.

[0675] 2.3 Dosage regimen:

[0676] Eight male Wistar rats were used in each group. After fasting overnight, the drug was orally administered at a dose of 1 mg / kg in a volume of 5 mL / kg.

[0677] The experiment was started when the rats weighed 200-230 g. Vehicle or drug was administered 1 h, 5 h, 11 h, 17 h, and 23 h before the administration of the modeling agent LPS. Samples were collected 1 h after the tail vein injection of the modeling agent LPS or PBS.

[0678] 2.4 Sample collection: Blood was collected from the jugular vein of rats, and the collected blood was added to a centrifuge tube, placed at room temperature for 60 minutes, and centrifuged at 8000 rpm for 5 minutes. 50 μL of the serum after centrifugation was transferred to a new labeled centrifuge tube, and 2 portions were taken, quickly frozen with dry ice, and stored in a -80°C refrigerator for testing.

[0679] 2.5 Sample processing: Serum samples: After thawing, the normal group and the 6-hour administration group were diluted 1.5 times, that is, 80.0 μL of sample was added to 40.0 μL 1x reagent diluent buffer (prepared by R&D kit); the modeling group and the 18-hour administration group were diluted 3 times, that is, 40.0 μL of sample was added to 80.0 μL 1x reagent diluent buffer.

[0680] 2.6 Experimental Procedure

[0681] TNF-α test (Kit DY510):

[0682] 1) Coat the test plate one day in advance: Take appropriate coating antibody and add it to PBS to obtain a coating antibody solution with a concentration of about 4.00 μg / mL, add it to the test plate, 100.0 μL per well, and place it in a 24°C incubator for overnight incubation.

[0683] 2) Washing: Discard the coating solution in the test plate, add 300.0ul of wash buffer to each well, and repeat washing 3 times, removing all residual liquid in the test plate each time.

[0684] 3) Blocking: Add 300.0 μL of 1× blocking buffer to each well for blocking and incubate at 24°C for at least 1 hour. 4) Prepare standard: First dilute the Rat TNFa standard to 1000 pg / ml, take 4.17 μL of the standard and add 495.83 μL of 1xreagent dilute, then dilute it twice with 1xRegent dilute to prepare 8 concentrations of standard.

[0685] 5) Take out the sealed test plate and wash the plate.

[0686] 6) Sample incubation: Take 100.0 μL of the diluted sample or standard into the corresponding test well of the ELISA test plate, seal it with adhesive tape and place it in a 24°C incubator for 2 hours.

[0687] 7) Wash the plate.

[0688] 8) Detection antibody incubation: Prepare the detection antibody working solution: Add appropriate detection antibody to 1xRegent Dilute to obtain the detection antibody solution. Add 100.0 μL of detection antibody working solution to each well, seal with new adhesive tape, and incubate in a 24°C incubator for 2 hours.

[0689] 9) Wash the plate.

[0690] 10) Secondary antibody HRP incubation in dark: Take appropriate amount of 40xStreptavidin-HRP and dilute it to 1x in 1xRegent Dilute. Add 100.0μL to each well and incubate in 24℃ incubator for about 20min.

[0691] 11) Wash the plate and repeat step 2.

[0692] 12) Color development and light protection: Mix equal volumes of color reagent A and B to prepare substrate solution, add 100.0ul to each well, and incubate at room temperature for about 20 minutes.

[0693] 13) Stop: When the color development is appropriate, add 50.0 μL STOP solution to each well.

[0694] IL-6 detection (Kit K15059D-2):

[0695] 1) Reagent preparation: Preparation: Take out the reagent kit from the refrigerator and equilibrate it at room temperature for 30 minutes.

[0696] 2) Prepare the standard: add 1 ml of diluent 42 to the standard, shake thoroughly, and leave at room temperature for 15-20 minutes. As the highest concentration point, make 4-fold dilutions in sequence, 7 standards + 1 Blank.

[0697] 3) Prepare samples: dilute rat serum appropriately

[0698] 4) Prepare the detection antibody diluent: aspirate the detection antibody and make up the volume with diluent 40.

[0699] 5) Prepare the cleaning solution: prepare 1*PBS (0.05% Tween-20) for later use.

[0700] 6) Prepare plate reading solution: Prepare 2* plate reading solution. (Prepare 10 minutes before the end of antibody incubation)

[0701] 7) Take out the rewarmed MSD plate and Blocker H, add 150ul Block H to each well and incubate at room temperature with shaking at 800rpm for 1h.

[0702] 8) Wash the MSD plate 3 times, each time with 150ul of washing solution.

[0703] 9) Add 50ul of diluted samples and standards to each well, seal with sealing film, and shake at room temperature for 2h.

[0704] 10) Wash the MSD plate 3 times, each time with 150ul of washing solution.

[0705] 11) Add 25ul of the prepared detection antibody to each well, seal with sealing film, and shake at room temperature for 2 hours.

[0706] 12) Wash the MSD plate 3 times, each time with 150ul of washing solution.

[0707] 13) Add 150ul of prepared plate reader solution to each well and test on the machine.

[0708] 3 Experimental results and conclusions:

[0709] Data processing was performed using Graph pad; the results of the PD experiment on the Wistar rat acute inflammation model showed that the compound of the present invention exhibited an excellent PD effect and had a high inhibition rate on both TNF-α and IL-6.

[0710] Crystalline Form Study of Examples

[0711] It is well known to those skilled in the art that the above-mentioned example compounds have been shown to have a good inhibitory effect on the expression level of TNF-α in U937 cells, and their crystal forms often have the same pharmacological and pharmacodynamic activity. On this basis, the inventors further studied the physical and chemical properties of the crystal forms of the corresponding compounds, but the preparation and characterization of the following specific crystal forms do not represent a limitation on the scope of protection of the present invention. Those skilled in the art can obtain more crystals of the compounds of the present invention based on the present invention by conventional crystallization methods, and these crystals are all the solutions protected by the present invention. Specifically as follows:

[0712] 1.1 Experimental Instruments

[0713] 1.1.1 Some parameters of physical and chemical testing instruments

[0714]

[0715]

[0716] 1.2 Instruments and liquid phase analysis conditions

[0717] 1.2.1 Instruments and equipment

[0718] Analytical balance (Sartorius BSA224S-CW), water purifier (Milli-Q Plus, Millipore), high performance liquid chromatograph (Agilent1260), pump (Agilent G1311B), injector (G1329B), column oven (G1316A), detector (G1316A).

[0719] 1.2.2 Chromatographic conditions

[0720] Chromatographic column: ZORBAX SB C8 (4.6*75mm, 3.5μm)

[0721] Flow rate: 1.0mL / min

[0722] Column temperature: 35°C

[0723] Detection wavelength: 254nm

[0724] Injection volume: 3 μL

[0725] Run time: 10 minutes

[0726] Diluent: DMSO

[0727] Mobile phase: A: water (0.05% trifluoroacetic acid); B: acetonitrile (0.05% trifluoroacetic acid)

[0728] T(min) A(％) B(％) 0.00 90.0 10.0 4.00 20.0 80.0 7.00 20.0 80.0 7.01 90.0 10.0 10.00 90.0 10.0

[0729] 1. Preparation of different crystal forms of the compound of Example 1

[0730] 1.1 Preparation of Form A

[0731] Weigh 10 mg of the free base amorphous form, add 50 μL of acetone, precipitate after slight turbidity, slurry at 50°C for 1 day, centrifuge, remove the supernatant, and dry the solid at 40°C in vacuum. Figure 1 The XRPD pattern shown in Figure 2 The DSC graph shown is Form A.

[0732] 1.2 Preparation of Form A

[0733] Weigh 10 mg of the free base amorphous form, add 50 μL of methanol, dissolve and precipitate, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and XRPD is detected to be Form A.

[0734] 1.3 Preparation of Form A

[0735] Weigh 10 mg of the free base amorphous form, add 50 μL of ethanol, dissolve and precipitate, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and then detected by XRPD to be Form A.

[0736] 1.4 Preparation of Form A

[0737] Weigh 10 mg of the free base amorphous form, add 50 μL of dichloromethane, dissolve and precipitate, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and XRPD is detected to be Form A.

[0738] 1.5 Preparation of Form A

[0739] Weigh 10 mg of the free base amorphous form, add 50 μL of acetonitrile, dissolve and precipitate, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and then detected by XRPD to be Form A.

[0740] 1.6 Preparation of Form A

[0741] Weigh 10 mg of the free base amorphous form, add 50 μL of 1,4-dioxane, dissolve and precipitate, slurry at 50°C for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40°C and XRPD is detected to be Form A.

[0742] 1.7 Preparation of Form A

[0743] Weigh 10 mg of the free base amorphous form, add 50 μL of isopropanol, dissolve and precipitate, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and XRPD is detected to be Form A.

[0744] 1.8 Preparation of Form A

[0745] Weigh 10 mg of the free base amorphous form, add 50 μL of 2-butanone, dissolve and precipitate, slurry at 50°C for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40°C and XRPD is detected to be Form A.

[0746] 1.9 Preparation of Form A

[0747] Weigh 10 mg of the free base amorphous form, add 50 μL of isopropyl acetate, precipitate after slight turbidity, slurry at 50° C. for 1 day, centrifuge, remove the supernatant, and the obtained solid is vacuum dried at 40° C. and XRPD is detected to be Form A.

[0748] 1.10 Preparation of Form A

[0749] Weigh 10 mg of the free base amorphous form, add 40 μL of acetone, sonicate to dissolve it, add 200 μL of the anti-solvent methyl tert-butyl ether to precipitate a solid, centrifuge, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form A.

[0750] 1.11 Preparation of Form B

[0751] Weigh 10 mg of the free base amorphous form, add 50 μL of ethyl acetate, beat at 50°C for 1 day, centrifuge, remove the supernatant, and dry the obtained solid in vacuum at 40°C. Figure 3 The XRPD pattern shown in Figure 4 The DSC graph shown is Form B.

[0752] 1.12 Preparation of Form B

[0753] Weigh 10 mg of the free base amorphous form, add 40 μL of acetone, sonicate to dissolve it, add 100 μL of anti-solvent water to precipitate solid, centrifuge, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form B.

[0754] 1.13 Preparation of Form C

[0755] Weigh 100 mg of the free base amorphous form, add 1 mL of 3-pentanone, beat at 50°C for 1 day, vacuum filter, and dry the filter cake at 40°C to obtain a solid. Figure 5 The XRPD pattern shown is Form C.

[0756] 1.14 Preparation of Form D

[0757] Weigh 10 mg of the free base amorphous form, add 40 μL of acetonitrile, sonicate to dissolve it, add 100 μL of anti-solvent water to precipitate solid, centrifuge immediately, remove the supernatant, and dry the solid at 40°C in vacuum. Figure 6 The XRPD pattern shown is Form D.

[0758] 1.15 Preparation of Form D

[0759] Weigh 10 mg of the free base amorphous form, add 40 μL of acetonitrile, sonicate to dissolve it, add 100 μL of the anti-solvent methyl tert-butyl ether, solid precipitates, centrifuge immediately, remove the supernatant, and the obtained solid is vacuum dried at 40°C and XRPD is detected to be Form D.

[0760] 1.16 Preparation of Form D

[0761] Weigh 10 mg of the free base amorphous form, add 40 μL of acetonitrile, sonicate to dissolve it, add 100 μL of the anti-solvent n-hexane to precipitate a solid, immediately centrifuge, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form D.

[0762] 1.17 Preparation of Form D

[0763] Weigh 10 mg of the free base amorphous form, add 60 μL of dichloromethane, sonicate to dissolve it, add 100 μL of anti-solvent water to precipitate solid, immediately centrifuge, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form D.

[0764] 1.18 Preparation of Form D

[0765] Weigh 10 mg of the free base amorphous form, add 60 μL of dichloromethane, sonicate to dissolve it, add 200 μL of anti-solvent methyl tert-butyl ether to precipitate a solid, centrifuge immediately, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form D.

[0766] 1.19 Preparation of Form D

[0767] Weigh 10 mg of the free base amorphous form, add 60 μL of dichloromethane, sonicate to dissolve it, add 100 μL of anti-solvent n-hexane to precipitate solid, centrifuge immediately, remove the supernatant, and dry the obtained solid in vacuum at 40°C. XRPD detection shows that it is Form D.

[0768] 1.20 Preparation of Form E

[0769] Weigh 100 mg of the free base amorphous, add 1 mL of water, slurry at 50°C for 4 days, vacuum filter, and vacuum dry the filter cake at 40°C to obtain a solid. Figure 7 The XRPD pattern shown is Form E.

[0770] 2. Solid stability test

[0771] 2.1 Experimental Purpose:

[0772] The physicochemical stability of the crystalline compound of the present invention was investigated under high temperature 60°C, room temperature and high humidity RH=92.5% and 50°C RH=75% for 7 days and 14 days.

[0773] 2.2 Experimental plan:

[0774] About 1 mg of the crystalline form of the present invention was taken and placed in an oven at 60°C, room temperature and high humidity RH=92.5% and 50°C RH=75% for 7 days and 14 days, and the changes of related substances were calculated using the HPLC peak area normalization method.

[0775] 2.3 Experimental results:

[0776] The physicochemical stability results of Form A under different conditions:

[0777] Table 6

[0778]

[0779]

[0780] From the stability data, it can be seen that Form A is relatively stable under high temperature, high humidity and high temperature and high humidity conditions, and no significant increase in impurities is observed.

[0781] 3. Hygroscopicity test

[0782] 3.1 Experimental Purpose

[0783] The hygroscopicity of the crystalline form of the compound of the present invention under different relative humidity conditions was investigated.

[0784] 3.2 Experimental plan:

[0785] The crystal form of the compound of the present invention is placed in saturated water vapor with different relative humidity to allow the crystal form and the water vapor to reach dynamic equilibrium, and the percentage of moisture absorption and weight gain of the crystal form after equilibrium is calculated.

[0786] 3.3 Experimental results:

[0787] Crystalline Form A absorbs moisture at RH80% and gains weight by about 1.059%, which is slightly hygroscopic. After 2 cycles of moisture absorption and desorption at 0-95% relative humidity, the XRPD spectrum of Crystalline Form A does not change, that is, the crystal form does not change.

Claims

1. A crystalline form of a compound represented by general formula (I) or a stereoisomer thereof, in: X1 is selected from N or CR 1 ; R 1 is selected from hydrogen or halogen; R 2 Selected from hydrogen, C 1-3 Alkyl or halogen; R 3 Selected from C 1-3 Alkyl, C 1-3 Deuterated alkyl or C 3-6 Cycloalkyl; R aa is selected from hydrogen or deuterium; and R bb Selected from hydrogen or deuterium.

2. The crystalline form of the compound of formula (I) or its stereoisomer according to claim 1, characterized in that: The compound is selected from: (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((2,4-difluorophenyl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-5',6-dimethyl-2-carbonyl-4-((2,4,6-trifluorophenyl)methoxy-d2)-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)propanamide; (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide-2,2,2-d3; (S)-N-(2-(4-chloro-1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-1H-pyrazol-3-yl)propan-2-yl)acetamide; (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-oxo-2H-[1,4'-bipyridyl]-2'-yl)-4-methyl-1H-pyrazol-3-yl)propan-2-yl)acetamide; or (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)cyclopropanecarboxamide.

3. The crystalline form of the compound or its stereoisomer according to claim 2, characterized in that: The crystalline form of the compound (S)-N-(2-(1-(3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy-d2)-5',6-dimethyl-2-carbonyl-2H-[1,4'-bipyridyl]-2'-yl)-4-fluoro-1H-pyrazol-3-yl)propan-2-yl)acetamide is Form A, Form B, Form C, Form D or Form E; in: The X-ray powder diffraction spectrum of the crystalline form A has a diffraction peak at 2θ of 5.3±0.2°; or a diffraction peak at 9.4±0.2°; or a diffraction peak at 10.7±0.2°; or a diffraction peak at 12.3±0.2°; or a diffraction peak at 15.9±0.2°; or a diffraction peak at 16.8±0.2°; or a diffraction peak at 17.4±0.2°; or a diffraction peak at 19.1±0.2°; or a diffraction peak at 22.2±0.2°; or a diffraction peak at 26.8±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; The X-ray powder diffraction spectrum of the crystalline form B has a diffraction peak at 2θ of 5.2±0.2°; or a diffraction peak at 8.0±0.2°; or a diffraction peak at 10.1±0.2°; or a diffraction peak at 10.4±0.2°; or a diffraction peak at 11.4±0.2°; or a diffraction peak at 12.0±0.2°; or a diffraction peak at 12.7±0.2°; or a diffraction peak at 15.5±0.2°; or a diffraction peak at 17.8±0.2°; or a diffraction peak at 19.1±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; The X-ray powder diffraction spectrum of the crystalline form C has a diffraction peak at 2θ of 7.3±0.2°; or a diffraction peak at 8.6±0.2°; or a diffraction peak at 9.2±0.2°; or a diffraction peak at 11.7±0.2°; or a diffraction peak at 15.2±0.2°; or a diffraction peak at 17.2±0.2°; or a diffraction peak at 19.8±0.2°; or a diffraction peak at 21.6±0.2°; or a diffraction peak at 22.3±0.2°; or a diffraction peak at 24.2±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; The X-ray powder diffraction spectrum of the crystalline form D has a diffraction peak at 2θ of 7.2±0.2°; or a diffraction peak at 8.6±0.2°; or a diffraction peak at 9.3±0.2°; or a diffraction peak at 11.8±0.2°; or a diffraction peak at 15.2±0.2°; or a diffraction peak at 15.6±0.2°; or a diffraction peak at 18.7±0.2°; or a diffraction peak at 20.1±0.2°; or a diffraction peak at 21.5±0.2°; or a diffraction peak at 24.5±0.2°; preferably including any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above diffraction peaks, more preferably including any 6, 7 or 8 thereof; The X-ray powder diffraction pattern of the crystalline form E has a diffraction peak at 2θ of 4.4±0.2°; or a diffraction peak at 10.8±0.2°; or a diffraction peak at 11.6±0.2°; or a diffraction peak at 12.6±0.2°; or a diffraction peak at 13.0±0.2°; or a diffraction peak at 13.3±0.2°; or a diffraction peak at 15.3±0.2°; or a diffraction peak at 16.9±0.2°; or a diffraction peak at 21.0±0.2°; or a diffraction peak at 25.4±0.2°; preferably includes any 2-5, or 3-5, or 3-6, or 3-8, or 5-8, or 6-8 of the above-mentioned diffraction peaks, and more preferably includes any 6, 7 or 8 thereof.

4. The crystalline form of the compound or its stereoisomer according to claim 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form A comprises at least one or more diffraction peaks located at 2θ of 10.7±0.2°, 15.9±0.2°, and 17.4±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 5.3±0.2°, 9.4±0.2°, 12.3±0.2°, 19.1±0.2°, and 22.2±0.2°, preferably comprising 2, 3, 4 or 5; optionally further comprising one or more diffraction peaks located at 2θ of 13.4±0.2°, 13.9±0.2°, 16.8±0.2°, 18.8±0.2°, 22.6±0.2°, 26.8±0.2°, 28.6±0.2°; preferably comprising at least any 2-3, or 4-5, or 6-7 thereof; further preferably, comprising any 2, 3, 4, 5, 6, 7 thereof; The X-ray powder diffraction pattern of the crystalline form B at least comprises one or more diffraction peaks located at 2θ of 10.4±0.2°, 11.4±0.2°, and 15.5±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 12.0±0.2°, and 17.8±0.2°, preferably comprising 2, 3, 4 or 5; optionally further comprising one or more diffraction peaks located at 2θ of 12.2±0.2°, 12.7±0.2°, 16.9±0.2°, 17.5±0.2°, 19.1±0.2°, 21.7±0.2°, 25.1±0.2°; preferably comprising at least any 2-3, or 4-5, or 6-7 thereof; further preferably, comprising any 2, 3, 4, 5, 6, 7 thereof; The X-ray powder diffraction pattern of the crystalline form C comprises at least one or more diffraction peaks located at 2θ of 7.3±0.2°, 8.6±0.2°, and 15.2±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 9.2±0.2°, 11.7±0.2°, 21.6±0.2°, 22.3±0.2°, and 24.2±0.2°, preferably 2 of them. , 3, 4 or 5; optionally further comprising one or more diffraction peaks located at 2θ of 15.6±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 27.6±0.2°, 27.8±0.2°, 37.0±0.2°; preferably comprising at least any 2-3, or 4-5, or 6-7 thereof; further preferably, comprising any 2, 3, 4, 5, 6, 7 thereof; The X-ray powder diffraction pattern of the crystalline form D comprises at least one or more diffraction peaks located at 2θ of 7.2±0.2°, 9.3±0.2°, and 15.2±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 8.6±0.2°, 11.8±0.2°, 18.7±0.2°, 20.1±0.2°, and 24.5±0.2°, preferably 2 of them. , 3, 4 or 5; optionally further comprising one or more diffraction peaks located at 2θ of 12.4±0.2°, 15.6±0.2°, 17.2±0.2°, 21.5±0.2°, 23.7±0.2°, 25.4±0.2°, 28.2±0.2°; preferably comprising at least any 2-3, or 4-5, or 6-7 thereof; further preferably, comprising any 2, 3, 4, 5, 6, 7 thereof; The X-ray powder diffraction pattern of the crystalline form E at least comprises one or more diffraction peaks located at 2θ of 4.4±0.2°, 13.3±0.2°, and 16.9±0.2°, preferably two of them, and more preferably three of them; optionally, it may further comprise at least one of 2θ of 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 15.3±0.2°, and 25.4±0.2°, preferably comprising 2, 3, 4 or 5; optionally further comprising one or more diffraction peaks located at 2θ of 13.0±0.2°, 14.2±0.2°, 17.5±0.2°, 18.7±0.2°, 21.0±0.2°, 21.9±0.2°, 26.2±0.2°; preferably comprising at least any 2-3, or 4-5, or 6-7 thereof; further preferably, comprising any 2, 3, 4, 5, 6, or 7 thereof.

5. The crystalline form of the compound or its stereoisomer according to claim 3, characterized in that: The X-ray powder diffraction pattern of the crystalline form A comprises one or more diffraction peaks located at 2θ of 5.3±0.2°, 9.4±0.2°, 10.7±0.2°, 12.3±0.2°, 13.4±0.2°, 15.9±0.2°, 16.8±0.2°, 17.4±0.2°, 18.8±0.2°, 19.1±0.2°, 22.2±0.2°, and 26.8±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 of them; The X-ray powder diffraction pattern of the crystalline form B comprises one or more diffraction peaks located at 2θ of 5.2±0.2°, 8.0±0.2°, 10.1±0.2°, 10.4±0.2°, 11.4±0.2°, 12.0±0.2°, 12.2±0.2°, 12.7±0.2°, 15.5±0.2°, 16.9±0.2°, 17.8±0.2°, and 19.1±0.2°; preferably, it comprises 4, 5, 6, 8, 10, or 12 of them; The X-ray powder diffraction pattern of the crystalline form C comprises one or more diffraction peaks located at 2θ of 7.3±0.2°, 8.6±0.2°, 9.2±0.2°, 11.7±0.2°, 15.2±0.2°, 17.2±0.2°, 18.5±0.2°, 19.8±0.2°, 21.6±0.2°, 22.3±0.2°, 24.2±0.2°, and 27.8±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 of them; The X-ray powder diffraction pattern of the crystalline form D comprises one or more diffraction peaks located at 2θ of 7.2±0.2°, 8.6±0.2°, 9.3±0.2°, 11.8±0.2°, 15.2±0.2°, 15.6±0.2°, 17.2±0.2°, 18.7±0.2°, 20.1±0.2°, 21.5±0.2°, 24.5±0.2°, and 28.2±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 of them; The X-ray powder diffraction pattern of the crystalline form E comprises one or more diffraction peaks located at 2θ of 4.4±0.2°, 10.8±0.2°, 11.6±0.2°, 12.6±0.2°, 13.0±0.2°, 13.3±0.2°, 14.2±0.2°, 15.3±0.2°, 16.9±0.2°, 21.0±0.2°, 21.9±0.2°, and 25.4±0.2°; preferably, it comprises 4, 5, 6, 8, 10 or 12 selected therefrom.

6. The compound according to any one of claims 3 to 5 or a crystalline form of its stereoisomer, characterized in that: The 2θ error between the diffraction peak positions with the top ten relative peak intensities in the X-ray powder diffraction pattern of the crystalline form A, form B, form C, form D or form E and the diffraction peaks at the corresponding positions in Figures 1, 3, 5, 6 or 7 is ±0.2° to ±0.5°, preferably ±0.2° to ±0.3°, and most preferably ±0.2°.

7. The compound according to any one of claims 3 to 6 or a crystalline form of its stereoisomer, characterized in that: The X-ray powder diffraction pattern of the crystalline form A is shown in FIG1 ; and / or the crystalline form A has a DSC pattern as shown in FIG2 ; The X-ray powder diffraction pattern of the crystalline form B is shown in FIG3 ; and / or the crystalline form B has a DSC pattern as shown in FIG4 ; The X-ray powder diffraction pattern of the crystalline form C is shown in FIG5 ; The X-ray powder diffraction pattern of the crystal form D is shown in FIG6 ; The X-ray powder diffraction pattern of the crystalline form E is shown in FIG7 .

8. A method for preparing a crystalline form of the compound or its stereoisomer according to any one of claims 1 to 7: Method 1 comprises the following steps: the compound is slurried with a solvent at a certain temperature to obtain a crystalline form, preferably, the temperature is 0 to 50°C; more preferably, the temperature is 20 to 30°C; The solvent is selected from one or more of water, methanol, ethanol, isopropyl acetate, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol n-propanol, tert-butanol, 2-butanone, 3-pentanone and N-methylpyrrolidone; preferably one or more of water, methanol, ethanol, isopropyl acetate, acetone, ethyl acetate, acetonitrile, dichloromethane, 1,4-dioxane, isopropanol, 2-butanone or 3-pentanone; Method 2 comprises the following steps: dissolving the compound in a good solvent, adding a poor solvent and stirring until a solid is precipitated to obtain a crystalline form; The good solvent is selected from one or more of ethyl acetate, isopropyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran or 1,4-dioxane; preferably one or more of acetone, dichloromethane or acetonitrile; The poor solvent is selected from one or more of n-heptane, cyclohexane, n-hexane, n-pentane, water, ethyl acetate, methyl tert-butyl ether, toluene or isopropyl ether; preferably one or more of water, n-hexane or methyl tert-butyl ether.

9. A pharmaceutical composition comprising a therapeutically effective dose of the compound according to any one of claims 1 to 7 or a crystalline form of its stereoisomer and one or more pharmaceutically acceptable carriers, diluents or excipients.

10. The pharmaceutical composition according to claim 9, wherein The weight percentage of the crystalline form of the compound or its stereoisomer is 0.1% to 95%, preferably 0.5% to 85%, more preferably 1% to 60%, further preferably 10% to 50%, further preferably 15-40%, further preferably 20-30%, further preferably 20-25%.

11. Use of the compound according to any one of claims 1 to 7 or the crystal form of its stereoisomer, or the pharmaceutical composition according to claim 9, in the preparation of a drug for treating a disease mediated by p38 kinase.

12. Use of the compound according to any one of claims 1 to 7 or the crystalline form of its stereoisomer, or the pharmaceutical composition according to claim 9, in the preparation of a medicament for treating autoimmune diseases, chronic inflammatory diseases, acute inflammatory conditions, autoinflammatory diseases, atherosclerosis, diabetes, fibrotic diseases, metabolic diseases, cancers, tumors, leukemia and lymphoma.

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