A crystal form of a terpyridine diketone compound represented by formula (I) and a preparation method and application thereof

Abstract

The application belongs to the technical field of chemical medicines and relates to a crystal form of a terpyridine diketone compound shown in formula (I), which is a p38 / MK2 inhibitor, can inhibit the production of a cytokine TNF alpha and is used for treating diseases such as arthritis.

Description

Technical Field

[0001] This invention belongs to the field of chemical pharmaceutical technology, and relates to a class of terpyridinedione compounds, their preparation methods and applications. Background Technology

[0002] Mitogen-activated protein kinases (MAPKs) are a conserved family of enzymes that use phosphorylation cascades to transmit and deliver external stimuli in order to produce coordinated cellular responses to the environment. MAPKs are proline-guided serine / threonine-specific protein kinases that regulate cellular activities such as gene expression, mitosis, differentiation, and cell survival / apoptosis. To date, four distinct classes of mammalian MAPKs have been identified: extracellular signal transduction kinases (ERK1 and ERK2), c-jun N-terminal kinase-1 (JNK1-3), p38MAPKs (p38α, p38β, p38γ, and p38δ), and ERK5.

[0003] Scientific investigation of this pathway from a biological, cellular, and in vivo perspective is primarily achieved through the availability of highly effective, selective small-molecule inhibitors targeting the α-isomer of p38MAPK and, to a lesser extent, the β-isomer. p38αMAPK is a major isomer involved in immune and inflammatory responses. Therefore, its function is crucial for the production and activity of various pro-inflammatory cytokines in cells such as macrophages, monocytes, synovial cells, and endothelial cells, including TNFα, IL-1, IL-6, and IL-8. p38MAPK is also responsible for inducing key inflammatory enzymes, such as COX2 and iNOS, which are major sources of arachidonic acid and nitric oxide at inflammatory sites, respectively. Furthermore, the p38MAPK pathway regulates the expression of matrix metalloproteinases (MMPs), including MMP2, MMP9, and MMP13.

[0004] The use of selective and effective inhibitors has facilitated the discovery of several families of p38MAPK substrates, including transcription factors, MAPKAP kinases, and other enzymes. MAPKAP kinases (MK2, MK-3, and PRAK) are selectively phosphorylated by p38MAPK, while phosphorylation of MSK1 / 2, MNK1 / 2, and RSKb is catalyzed by both p38MAPK and ERK. Although substrate identification is challenging due to the lack of specific inhibitors, RSKb activation is thought to play a role in cell survival.

[0005] Once phosphorylated and activated by p38MAPK, MK-2, MK-3, and PRAK share similar substrate specificity. All of these kinases phosphorylate the small heat shock protein Hsp27. Studies have shown that PRAK- and MK3-deficient mice do not exhibit any tolerance to endotoxin shock or lipopolysaccharide (LPS)-induced reductions in cytokine production. In contrast, MK-2-deficient mice show tolerance to endotoxin shock and impaired inflammatory responses, as well as a significant reduction in the production of cytokines such as TNFα, IFNγ, and IL-6. Therefore, the p38 / MK2 axis is particularly necessary and sufficient for regulating pro-inflammatory responses.

[0006] By utilizing the p38:MK2 interaction and using MK2 as a p38 substrate, a novel p38α inhibitor exhibiting properties of interest was discovered (Davidson et al.). This inhibitor demonstrates substrate selectivity by preventing p38α-dependent phosphorylation of MK2 (Ki app 300 nM) while preserving p38α-dependent phosphorylation of ATF2 (Ki app > 20 μM). This novel inhibitor functions uniquely compared to conventional p38ATP-competitive inhibitors that block p38-dependent phosphorylation of all p38 substrates. A second independent study also described a p38 inhibitor with unique mechanistic properties. This work demonstrated a novel mechanism for selectively inhibiting p38-dependent phosphorylation of MK2. Unlike previous studies by Davidson et al., these compounds with unique mechanisms compete with ATP and stabilize the p38 / MK2 complex.

[0007] In summary, these two studies clearly demonstrate the concept that selective p38 / MK2 axis blockade can be achieved using small molecule inhibitors. Compared to conventional p38MAPK inhibitors, these p38 / MK2 inhibitors should retain or enhance efficacy and exhibit improved safety profiles in animal models of disease or in human clinical settings.

[0008] In its previous research, the applicant obtained a terpyridinedione compound of formula (I) which has a good p38MAPK inhibitory effect. The present invention further studies and develops the terpyridinedione compound to obtain a suitable pharmaceutical physical form. Summary of the Invention

[0009] The terpyridinedione compound crystal form provided in this application, as a p38 / MK2 inhibitor, can inhibit the production of cytokine TNFα, thereby regulating inflammatory responses and related diseases.

[0010] This application provides a polymorphic AO of the tripyridinedione compound represented by formula (I). Specifically, the present invention is achieved through the following technical solution:

[0011] In a first aspect, this application provides a crystalline form A of a terpyridinedione compound of formula (I), wherein,

[0012]

[0013] The X-ray diffraction pattern of crystal form A shows characteristic peaks at 5.00°, 7.70°, 12.82°, 14.08°, 15.44°, 16.72°, 17.17°, 19.70°, 20.41°, 20.88°, 23.08°, 23.78°, and 26.83°, with an error of ±0.2°.

[0014] Furthermore, the X-ray diffraction pattern of crystal form A, represented by the 2θ angle, also includes angles of 9.24°, 11.40°, 11.63°, 12.23°, 13.85°, 14.80°, 15.71°, 15.96°, 17.75°, 17.93°, 18.20°, 18.67°, 18.94°, 21.15°, 21.48°, 22.51°, and 22.71°. Characteristic peaks are observed at 22.44°, 24.65°, 25.20°, 25.66°, 25.84°, 26.60°, 27.47°, 27.74°, 27.92°, 28.81°, 29.57°, 30.27°, 30.72°, 31.38°, 32.11°, 32.70°, 33.57°, and 36.70°, with an error of ±0.2°.

[0015] Secondly, this application also provides a crystalline form B of the terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form B has characteristic peaks at 5.97°, 7.80°, 9.26°, 11.57°, 14.41°, 15.17°, 17.93°, 18.12°, 19.64°, 20.10°, 22.88°, and 23.50°, with an error of ±0.2°.

[0016] Thirdly, this application also provides a crystalline form C of the terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form C has characteristic peaks at 6.03°, 14.45°, 16.31°, 17.44°, 18.16°, 21.02°, 24.40°, 24.98°, 26.48°, and 27.84°, with an error of ±0.2°.

[0017] Fourthly, this application also provides a crystalline form D of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form D has characteristic peaks at 4.15° and 18.37°, indicated by an angle of 2θ, with an error of ±0.2°.

[0018] Fifthly, this application also provides a crystalline form E of the terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form E has characteristic peaks at 9.22°, 10.72°, 12.58°, 14.72°, 16.14°, 16.78°, 17.40°, 18.45°, 19.73°, 20.16°, 20.73°, 21.33°, 23.21°, 24.98°, 25.25°, and 27.82°, with an error of ±0.2°.

[0019] Sixthly, this application also provides a crystalline form F of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form F has characteristic peaks at 12.75°, 15.13°, 15.37°, 16.01°, 16.21°, 17.35°, 18.44°, and 19.30°, indicated by an angle of 2θ, with an error of ±0.2°.

[0020] Furthermore, the X-ray diffraction pattern of the crystal form F also shows characteristic peaks at 5.03°, 5.34°, 6.85°, 10.05°, 10.86°, 12.95°, 13.52°, 14.26°, 18.20°, 20.17°, 22.01°, 22.73°, 23.61°, 24.82°, 25.55°, 26.05°, 26.32°, and 28.09°, with an error of ±0.2°.

[0021] Furthermore, the X-ray diffraction pattern of the crystal form F also shows characteristic peaks at 3.80°, 10.64°, 13.72°, 16.95°, 21.29°, 22.30°, 27.21°, and 30.13°, with an error of ±0.2°, indicated by a 2θ angle.

[0022] Furthermore, the differential thermal analysis spectrum of crystal form F shows a characteristic absorption peak at 129.4℃, with an error of ±5℃, as detailed in the attached figure. Figure 17 As shown.

[0023] In a seventh aspect, this application also provides a terpyridine dione compound crystal form G of formula (I), wherein the X-ray diffraction pattern of the crystal form G has characteristic peaks at 5.06°, 6.46°, 9.24°, 10.07°, and 15.09°, represented by 2θ angles, with an error of ±0.2°.

[0024] Eighthly, this application also provides a crystalline form H of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form H has characteristic peaks at 4.69°, 6.52°, 10.13°, 14.97°, 15.91°, 16.94°, 18.41°, 19.17°, 19.99°, 21.27°, and 22.49°, with an error of ±0.2°.

[0025] Ninthly, this application also provides a crystalline form I of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form I has characteristic peaks at 9.47°, 13.46°, 15.07°, 17.91°, 19.19°, 21.33°, and 25.39°, indicated by an angle of 2θ, with an error of ±0.2°.

[0026] In a tenth aspect, this application also provides a crystalline form J of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form J has characteristic peaks at 5.18°, 10.25°, 12.93°, 13.51°, 14.53°, 15.24°, 16.55°, 17.52°, 18.45°, 19.38°, 24.83°, 25.72°, and 28.01°, with an error of ±0.2°.

[0027] Eleventhly, this application also provides a crystalline form K of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form K has characteristic peaks at 15.28°, 17.75°, 19.54°, 19.93°, 20.80°, and 22.34°, indicated by an angle of 2θ, with an error of ±0.2°.

[0028] Furthermore, the X-ray diffraction pattern of the crystal form K, represented by a 2θ angle, also shows characteristic peaks at 5.00°, 7.49°, 11.40°, 12.80°, 14.02°, 15.56°, 16.43°, 16.84°, 17.07°, 18.10°, 18.70°, 18.90°, 21.29°, 22.65°, 23.70°, 23.87°, 24.36°, 24.59°, 25.04°, 25.76°, 26.48°, 27.29°, 27.84°, 28.27°, 29.47°, 30.05°, and 31.30°, with an error of ±0.2°.

[0029] Furthermore, the X-ray diffraction pattern of the crystal form K, represented by a 2θ angle, also shows characteristic peaks at 9.12°, 9.86°, 12.15°, 13.20°, 14.72°, 18.47°, 22.90°, 30.52°, 30.89°, 32.66°, 33.53°, 34.18°, 35.55°, 35.92°, 36.14°, 36.51°, 37.38°, 38.12°, 38.49°, 38.94°, 40.18°, 42.49°, and 42.92°, with an error of ±0.2°.

[0030] In a twelfth aspect, this application also provides a crystalline form L of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form L has characteristic peaks at 16.62°, 17.56°, and 21.60°, indicated by an angle of 2θ, with an error of ±0.2°.

[0031] In a thirteenth aspect, this application also provides a crystalline form M of a terpyridine dione compound represented by formula (I), wherein the X-ray diffraction pattern of the crystalline form M has characteristic peaks at angles of 6.42°, 9.24°, 10.06°, 13.55°, 13.71°, 15.05°, 15.67°, 16.49°, 17.52°, 18.06°, 18.51°, 19.13°, 20.08°, 21.25°, 22.53°, 24.42°, 25.31°, and 25.58°, with an error of ±0.2°.

[0032] In a fourteenth aspect, this application also provides a terpyridine dione compound crystal form N represented by formula (I), wherein the X-ray diffraction pattern of the crystal form N has characteristic peaks at 16.49° and 17.46°, indicated by a 2θ angle, with an error of ±0.2°.

[0033] Furthermore, the X-ray diffraction pattern of the crystal form N also shows characteristic peaks at 7.88°, 8.71°, 13.13°, 18.63°, 19.58°, 19.93°, 21.48°, and 22.96°, with an error of ±0.2°, indicated by a 2θ angle.

[0034] Furthermore, the X-ray diffraction pattern of the crystal form N also shows characteristic peaks at 11.45°, 14.10°, 15.19°, 15.34°, 17.97°, 20.78°, 22.55°, 23.27°, 23.91°, 24.42°, 24.96°, 25.29°, 26.32°, 27.18°, 28.44°, 28.81°, 29.12°, 29.47°, 30.72°, and 34.27°, with an error of ±0.2°.

[0035] In a fifteenth aspect, this application also provides a terpyridine dione compound crystal form O represented by formula (I), wherein the X-ray diffraction pattern of the crystal form O has characteristic peaks at 5.22°, 14.68°, 16.66°, 18.61°, 19.15°, 19.83°, and 23.41°, indicated by an angle of 2θ, with an error of ±0.2°.

[0036] This application further provides a pharmaceutical composition comprising a therapeutically effective amount of any of the crystal forms described above and one or more pharmaceutically acceptable carriers.

[0037] This application further provides the use of the crystal form in the preparation of a medicament for treating a disease, specifically a p38 / MK2-related disease, selected from chronic inflammatory diseases and acute inflammatory diseases, wherein the chronic inflammatory disease is preferably rheumatoid arthritis.

[0038] The terpyridinedione compound shown in formula (I) of this application has good stability and good purity, making it suitable for pharmaceutical applications.

[0039] For clarity, this article defines the general terminology used in the description of compounds.

[0040] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium capable of delivering an effective amount of the active substance of this invention without interfering with the biological activity of the active substance and without toxic side effects on the host or patient. Representative carriers include water, oil, vegetables and minerals, ointment bases, lotion bases, and ointment bases. These bases include suspending agents, thickeners, and transdermal penetration enhancers. Their formulations are well known to those skilled in the art of cosmetics or topical pharmaceuticals. For further information on carriers, see Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0041] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the quantity required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0042] The compounds of this application can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of this invention. Attached Figure Description

[0043] Figure 1 This is a schematic diagram of the crystal structure of a single molecule of compound 1A.

[0044] Figure 2 The crystal parameters are shown for the crystal structure of compound 1A.

[0045] Figure 3 The atomic coordinates and isotropic shift parameters of the crystal structure of compound 1A are shown.

[0046] Figure 4 The diagram shows the bond length parameters of the crystal structure of compound 1A.

[0047] Figure 5 The diagram shows the bond angle parameters of the crystal structure of compound 1A.

[0048] Figure 6 The diagram shows the dihedral angle parameters of the crystal structure of compound 1A.

[0049] Figure 7 This is the X-ray diffraction pattern of crystal form A as characterized for the first time.

[0050] Figure 8 The X-ray diffraction pattern is the result of the second characterization of crystal form A.

[0051] Figure 9 The X-ray diffraction pattern is for crystal form B.

[0052] Figure 10 The image shows the X-ray diffraction pattern of crystal form C.

[0053] Figure 11 The thermogravimetric analysis and differential thermal analysis spectra of crystal form C are shown.

[0054] Figure 12 This is the X-ray diffraction pattern of crystal form D.

[0055] Figure 13 The thermogravimetric analysis and differential thermal analysis spectra of crystal form D are shown.

[0056] Figure 14 This is the X-ray diffraction pattern of crystal form E.

[0057] Figure 15 The thermogravimetric analysis and differential thermal analysis spectra of crystal form E are shown.

[0058] Figure 16 The image shows the X-ray diffraction pattern of crystal form F prepared in one example.

[0059] Figure 17 The thermogravimetric analysis and differential thermal analysis spectra of crystal form F are shown.

[0060] Figure 18 X-ray diffraction pattern of crystal form F prepared in another embodiment.

[0061] Figure 19 This is the X-ray diffraction pattern of crystal form G.

[0062] Figure 20 This is the X-ray diffraction pattern of crystal form H.

[0063] Figure 21 The thermogravimetric analysis and differential thermal analysis spectra of crystal form H are shown.

[0064] Figure 22 This is the X-ray diffraction pattern of crystal form I.

[0065] Figure 23 The thermogravimetric analysis and differential thermal analysis spectra are for crystal form I.

[0066] Figure 24 The image shows the X-ray diffraction pattern of crystal form J.

[0067] Figure 25 This is the X-ray diffraction pattern of crystal form K as characterized for the first time.

[0068] Figure 26 This is the X-ray diffraction pattern of crystal form K after its second characterization.

[0069] Figure 27 This is the X-ray diffraction pattern of crystal form L.

[0070] Figure 28 This is the X-ray diffraction pattern of crystal form M.

[0071] Figure 29 The thermogravimetric analysis and differential thermal analysis spectra of crystal form M are shown.

[0072] Figure 30 This is the X-ray diffraction pattern of the first characterization of crystal form N. Figure 31 The thermogravimetric analysis and differential thermal analysis spectra of crystalline N are shown.

[0073] Figure 32 This is the X-ray diffraction pattern of the second characterization of crystal form N.

[0074] Figure 33 This is the X-ray diffraction pattern of crystal form O. Detailed Implementation

[0075] The present application will be described in further detail below with reference to the embodiments, but the implementation of the present application is not limited thereto.

[0076] Example 1

[0077]

[0078] The synthetic route for compound number 1 is as follows:

[0079] Step A: Synthesize 2-(bromomethyl)-3,5-difluoropyridine.

[0080]

[0081] At 0°C, triphenylphosphine (PPh3, 813.5 mg, 3.11 mmol) and carbon tetrabromide (CBr4, 823.8 mg, 2.48 mmol) were added sequentially to tetrahydrofuran (THF, 5.0 mL) containing (3,5-difluoro-2-pyridine)methanol (300.0 mg, 2.07 mmol), and the reaction was carried out at room temperature (rt) for 1 hour.

[0082] After the reaction was complete, the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1). 372.4 mg of a colorless oil, 2-(bromomethyl)-3,5-difluoropyridine, was obtained (yield: 86.5%). LC-MS: RT = 1.89 min, [M+H] + =208.01.

[0083] Step B: Synthesis of 3”-chloro-4”-((3,5-difluoropyridin-2-yl)methoxy)-3-(2-hydroxypropane-2-yl)-5',6”-dimethyl-2H,2”H-[1,2':4',1”-terpyridine]-2,2”-dione.

[0084]

[0085] At room temperature, 2-(bromomethyl)-3,5-difluoropyridine (37.4 mg, 0.18 mmol) and potassium carbonate (K2CO3, 33.12 mg, 0.24 mmol) were added to N,N-dimethylformamide (DMF, 2.0 mL) containing 3”-chloro-4”-hydroxy-3-(2-hydroxypropane-2-yl)-5',6”-dimethyl-2H,2”H-[1,2’:4’,1”-terpyridine]-2,2”-dione (50.0 mg, 0.12 mmol) and reacted at room temperature for 2 hours.

[0086] After the reaction was completed, the mixture was quenched with water, extracted with ethyl acetate (20 mL × 3 times), the organic phases were combined, washed with saturated brine (20 mL × 2 times), dried over anhydrous sodium sulfate, and concentrated. The residue was purified by silica gel column chromatography (eluent: ethyl acetate / n-hexane = 1 / 0). 28.0 mg of a white solid, 3”-chloro-4”-((3,5-difluoropyridin-2-yl)methoxy)-3-(2-hydroxypropane-2-yl)-5',6”-dimethyl-2H,2”H-[1,2':4',1”-terpyridine]-2,2”-dione, was obtained (yield: 53.7%).

[0087] LC-MS: RT = 1.86 min, [M+H] + =529.22. 1 H NMR (400MHz, DMSO) δ8.68(s,1H),8.60(d,J=2.3Hz,1H),8.13–8.05(m,1H),7.85(dd,J=6.9,2.1Hz,1H),7.78(s,1H),7.69(dd,J=7.0,2 .1Hz,1H),6.79(s,1H),6.42(t,J=7.0Hz,1H),5.47(d,J=1.6Hz,2H),5.22(s,1H),2.07(s,3H),2.00(s,3H),1.47(s,3H),1.46(s,3H).

[0088] The racemic compounds of Example 1 were separated using supercritical fluid chromatography (AS-H column) with a mobile phase of carbon dioxide and isopropanol, eluting successively the amorphous derivatives of Example 1A and 1B:

[0089]

[0090] Example 1A Compound: RT = 4.21 min (SFC, AS-H, 0.46 cm ID × 15 cm L column, 30% isopropanol isogradient method, flow rate 2.5 mL / min and circulation time 10 min); [a] D 25＝ –1.68° (MeOH, Rudolph Autopol I pyrolyte); 1H NMR (400MHz, DMSO) δ8.68(s,1H),8.59(d,J=2.3Hz,1H),8.11–8.05(m,1H),7.85(dd,J=6.9,2.0Hz,1H),7.79(s,1H),7.69(dd,J=7.0,2 .0Hz,1H),6.80(s,1H),6.42(t,J=6.9Hz,1H),5.48(d,J=1.2Hz,2H),5.23(s,1H),2.07(s,3H),2.00(s,3H),1.47(s,3H),1.46(s,3H).

[0091] Example 1B compound: RT = 4.68 min (SFC, AS-H, 0.46 cm ID × 15 cm L column, 30% isopropanol isogradient method, flow rate 2.5 mL / min and circulation time 10 min). [a] D 25 +1.64° (MeOH, Rudolph Autopol I pyrolyte); 1 H NMR (500MHz, DMSO) δ8.68(s,1H),8.59(d,J=2.3Hz,1H),8.11–8.06(m,1H),7.85(dd,J=6.9,2.1Hz,1H),7.78(s,1H),7.69(dd,J=7.0,2 .1Hz,1H),6.80(s,1H),6.42(t,J=6.9Hz,1H),5.47(d,J=1.3Hz,2H),5.24(s,1H),2.07(s,3H),2.00(s,3H),1.47(s,3H),1.46(s,3H).

[0092] Comparative Example 1

[0093] The structure of the compound 3-chloro-4-((3,5-difluoropyridin-2-yl)methoxy)-2'-(2-(2-hydroxypropyl-2-yl)pyrimidin-4-yl)-5',6-dimethyl-2H-[1,4'-bipyridin]-2-one is as follows:

[0094]

[0095] Synthetic routes are shown below. The synthetic route for compound 1 is described in Chinese Patent No. CN201480032278.5, specification number 49.

[0096] The racemic comparative example 1 compound was separated using supercritical fluid chromatography (OD-H column) with a mobile phase of carbon dioxide and ethanol, and the transisomers comparative example 1A and 1B were eluted sequentially.

[0097]

[0098] Comparative Example 1A: RT = 4.47 min (SFC, OD-H, 0.46 cm ID × 15 cm L column, 40% ethanol isogradient method, flow rate 2.5 mL / min and circulation time 10 min); [a] D 25 -0.66° (MeOH, Rudolph Autopol I pyrolyzer).

[0099] Comparative Example 1B compound: RT = 4.68 min (SFC, OD-H, 0.46 cm ID × 15 cm L column, 40% ethanol isogradient method, flow rate 2.5 mL / min and circulation time 10 min). [a] D 25 +0.68° (MeOH, Rudolph Autopol I pyrolyzer).

[0100] Example 2: LPS-induced TNFα release from U937 experiment

[0101] Cytokine Regulation in Human Monocytes: The p38 pathway has been shown to be key to the biosynthesis of various pro-inflammatory cytokines, including TNFα, IL-1β, and IL-6. Therefore, inhibition of the p38MAPK pathway reduces inflammatory responses by decreasing the biosynthesis of pro-inflammatory cytokines. This study demonstrates half the amount of the compound of the present invention required to inhibit the biosynthesis of TNFα (a pro-inflammatory cytokine). This reflects the effect of the compound of the present invention in reducing inflammation, which is beneficial for the treatment of many diseases, including chronic inflammatory conditions, acute inflammatory conditions, and autoinflammatory conditions. The efficacy and modality of the p38 inhibitor in blocking cytokine production were evaluated using the human U937 cell line.

[0102] Reagents and instruments:

[0103] 1640 medium, catalog number A10491-01, Gibco. Penicillin and streptomycin, catalog number 15140-122, Gibco. Fetal bovine serum, catalog number 10099-141C, Gibco. PBS, catalog number 10010-031, Gibco. LPS, catalog number L2880, Sigma. PMA, catalog number P1585, Sigma. Dimethyl sulfoxide, catalog number D8418-1L, Sigma. TNFα kit, catalog number K151QWD-4, MSD.

[0104] 96-well plate, item number 3599, Corning. Shaking plate shaker, item number QB-9002, Qilinbeier. Centrifuge, item number 5810R, Eppendorf. CO2 incubator, item number 371, Thermo. Counter, item number C10281, Gibco. Microscope, item number CKX41, OLYMPUS. MSD plate reader, 1201MESO SECTOR 600, MSD.

[0105] Experimental cells:

[0106] U937, ATCC, part number CRL-1593.2.

[0107] Drug preparation:

[0108] Weigh approximately 2 mg of the drug and prepare a 10 mM (based on free base) stock solution using DMSO. Dilute the stock solution 10-fold to 1 mM, then sequentially dilute it 4-fold to 250 μM, 62.5 μM, 15.6 μM, 3.9 μM, 0.97 μM, 0.24 μM, and 0.061 μM. Then, dilute each of these DMSO-concentrated drug solutions 20-fold with culture medium to prepare working solutions.

[0109] Experimental methods:

[0110] Day 0: Inoculate 10,000 cells / well, stimulate with 20 ng / ml PMA for 48 h, and incubate at 37°C and 5% CO2;

[0111] Day 2: 1. Remove the supernatant from the differentiated U937, wash once with PBS, and add 96 μl of 1640 medium;

[0112] 2. Add 2 μl of the compound containing (final concentration 0.1% DMSO), and incubate at 37°C with 5% CO2 for 30 min;

[0113] 3. Add 2 μl LPS (final concentration 100 ng / ml) to stimulate cells and incubate at 37°C and 5% CO2 for 4 h;

[0114] 4. Centrifuge, collect the supernatant, and use ELISA to determine the TNFα content in the supernatant.

[0115] Statistical methods:

[0116] The TNFα content for each well was calculated using the standard curve provided in the kit.

[0117] Example: Using GraphPad nonlinear fitting formula to calculate compound IC 50 The experimental results are shown in Table 1.

[0118] Table 1. IC50 of the compounds of this invention inhibiting TNFα production.50 value

[0119]

[0120] As can be seen from the experimental results in Table 1 above, the compounds of the present invention have significant inhibitory activity on TNFα production and can regulate inflammatory responses and related diseases.

[0121] Example 3: Determination of solubility.

[0122] 1. Preparation of control solution

[0123] Weigh approximately 0.5 mg of the sample to be tested into a centrifuge tube. First, add an appropriate amount of DMSO to completely dissolve the sample, and then add methanol to make up to 1 ml. The sample to be tested is compound number 1 and compound number 1 of comparative example.

[0124] 2. Preparation of the test solution

[0125] Weigh approximately 1.0 mg each of Comparative Example 1 and Compound No. 1 into two centrifuge tubes, and add 1 ml of PBS buffer solution (2.0 and 7.4) to each tube. (If multiple pH values ​​need to be investigated, the preparation method is similar).

[0126] 3. Place the prepared control solution and the test solution into a 37°C water bath and heat for 1 hour. After 1 hour, remove them and cool to room temperature. Filter the solution through a 0.22μm filter membrane before injection.

[0127] 4. Calculate the concentration of the sample in the test solution according to C=(A*(ms / vs)) / AS; the experimental results are shown in Table 2.

[0128] Note: ms, vs, and AS represent the weight, volume, and peak area of ​​the sample in the control solution, respectively.

[0129] A represents the peak area of ​​the test solution.

[0130] Table 2 shows the solubility of the compounds of this invention.

[0131]

[0132] As shown in Table 2, the compounds with application numbers 1, 2, 3 and 20 have excellent solubility, which is better than that of the compound in Comparative Example 1.

[0133] Example 4: p38α / MK2 complex kinase screening experiment

[0134] Table 3 Reagents:

[0135]

[0136] Table 4 Instruments:

[0137]

[0138] Drug preparation:

[0139] Weigh a certain amount of the drug and prepare a 10 mM stock solution with DMSO. Dilute the stock solution to 100 μM (100X working solution) in two steps, then dilute it 3-fold successively to 33.33 μM, 11.11 μM, 3.70 μM, 1.23 μM, 0.41 μM, 0.14 μM, 0.046 μM, 0.015 μM, and 0.005 μM. A DMSO control is also provided.

[0140] Then, the series of DMSO drugs diluted above were diluted 25 times with 1X IMAP Reaction Buffer to prepare 4X drug working solution.

[0141] Experimental methods:

[0142] a) Prepare the enzyme (MEK6 (active), p38α (unactive), MK2 (unactive), substrate (HSP27) and ATP using 1X IMAP Reaction Buffer.

[0143] b) In a black 384-well plate, add 10 μL of 2X enzyme & substrate working solution and 5 μL of 4X drug working solution sequentially. After rapid centrifugation, add 5 μL of ATP working solution to initiate the enzyme reaction. Each compound group has 10 concentration gradients, with 2 replicates. A DMSO control group and a negative control group (DMSO, no ATP) are also included. The final concentrations of each group are shown in Table 5 below.

[0144] Table 5

[0145]

[0146] c) After incubating at room temperature for 1 hour, add 1X Progressive Binding Solution to terminate the enzyme reaction. After incubating for 30 minutes, detect the fluorescence polarization (FP) signal [FP(Ex485 / Em520 / Em520nm)] using a BMGPHERASTER FSX multi-functional microplate reader.

[0147] Statistical methods:

[0148] Calculate the compound IC using a nonlinear fitting formula in GraphPad Prism 7. 50 The results are detailed in Table 6.

[0149] Table 6: Inhibition of IC50 by the compounds of the present invention on the p38α / MK2 complex 50 value

[0150]

[0151] As can be seen from the experimental results in Table 6 above, the compounds of the present invention have significant inhibitory activity against the p38α / MK2 complex and can regulate inflammatory responses and related diseases.

[0152] Example 5: p38α kinase screening experiment

[0153] Table 7 Reagents:

[0154]

[0155] Table 8 Instruments:

[0156]

[0157] Drug preparation:

[0158] Weigh a certain amount of the compound and prepare a 10 / 50mM stock solution with DMSO. Dilute the stock solution 3 times with DMSO to obtain 10 concentration points, which will be used as the working solution of the compound.

[0159] Experimental methods:

[0160] Transfer 50 nL of diluted compound working solution to each well of the reaction plate (784075, Greiner) using an Echo 655. Then add 2.5 μL (4 ng / μL) of p38α kinase solution and incubate at room temperature for 10 minutes. Next, add 2.5 μL of a mixture of kinase substrate (0.2 mg / mL) and ATP (150 μM) and react at room temperature for 60 minutes. Then add 4 μL of ADP Glo reagent and incubate at room temperature for 40 minutes. Finally, add 8 μL of kinase detection reagent and incubate at room temperature for 40 minutes. Read the luminescence signal using an Envision 2104.

[0161] Data Analysis

[0162] Calculate the compound IC using the nonlinear fitting formula of GraphPad Prism 8 50 The test results are shown in Table 9.

[0163] Table 9: Inhibition IC50 of the compounds of the present invention on p38α 50 value

[0164]

[0165] As can be seen from the experimental results in Table 9 above, compared with the prior art, the compound of the present invention has a higher activity fold relative to p38α / (p38α / MK2 complex) and better selectivity.

[0166] Note: p38α / MK2 complex kinase data are from Example 4.

[0167] Instruments used for crystal form characterization analysis

[0168] The XRD pattern and data of crystal form F were obtained using an Empyrean X-ray diffractometer with X-ray diffraction (XRD) spectra. The detection conditions were Cu-Kα radiation and wavelength. The divergence slit is 1 / 4°, the X-ray tube voltage is 45kV, the X-ray tube current is 40mA, the scanning range is 3-40° (2θ), the step size is 0.0262606°, and the dwell time per step is 33.915s.

[0169] The spectra and data of other crystal forms were obtained using a Bruker D8 Advance X-ray powder diffractometer (Bruker, GER). The scanning angle ranged from 2 to 45 degrees, with a scan step size of 0.02 μm and an exposure time of 0.12 seconds. The phototube voltage and current for the tested samples were 40 kV and 40 mA, respectively, and the sample disk was a zero-background sample disk.

[0170] Thermogravimetric analyzer (TGA): Model TA Discovery 550 (TA, US). Place 2-5 mg of sample in an aluminum sample pan, and then place another 2-5 mg of sample in a pre-equilibrated open aluminum sample pan. The samples are automatically weighed inside the TGA furnace. The samples are heated to the final temperature at a rate of 10 °C / min. The nitrogen purging rate at the sample location is 60 mL / min, and the nitrogen purging rate at the balance is 40 mL / min.

[0171] Differential scanning calorimetry (DSC): Model TA Discovery 250 (TA, US). 1-2 mg of sample was accurately weighed and placed in the perforated DSC Tzero sample pan. The sample was heated to the final temperature at a rate of 10 °C / min, and the nitrogen purging rate in the furnace was 50 mL / min.

[0172] High performance liquid chromatography (HPLC): model ACQUITY ARC-2489 (Waters, US).

[0173] Unless otherwise specified, the Pos. [°2θ] error of the crystal form of the present invention is ±0.2°.

[0174] Example 6: Preparation and characterization of crystal form A

[0175] 6.1 Preparation of Crystal Form A: 1.0036 g of compound 1A was added to 20.0 mL of ethanol / n-heptane (1 / 9, v / v) to form a suspension. The suspension was stirred at 50 °C for 1 day. The resulting solid was then vacuum dried overnight at 40 °C to obtain 0.9405 g of crystal form A (yield: 93.7%). A schematic diagram of the crystal structure of a single molecule is shown below. Figure 1 As shown, the crystal parameter diagram of the crystal structure is as follows: Figure 2 As shown, the atomic coordinates and isotropic displacement parameters of the crystal structure are plotted as follows: Figure 3 As shown, the bond length parameter diagram of the crystal structure is as follows: Figure 4 As shown, the bond angle parameter diagram of the crystal structure is as follows: Figure 5 As shown, the dihedral angle parameter diagram of the crystal structure is as follows: Figure 6 As shown in Figure 6.2. Characterization of Crystal Form A

[0176] The crystal form A obtained in section 6.1 above was used for XRD characterization. The specific characterization results are as follows: Figure 7 As shown in the table. The main characteristic absorption peaks of crystal form A are shown in Table 10:

[0177] Table 10 shows the XRD data for crystal form A.

[0178]

[0179] Crystal form A was prepared using the method described in Example 1 of section 6.1. Crystal form A was then used again for XRD characterization, and the specific characterization results are as follows. Figure 8 As shown in Table 11, the XRD characterization results of the two crystal forms A are compared, with an error of ±0.2°.

[0180] Table 11 shows the XRD data of crystal form A after two characterizations.

[0181]

[0182]

[0183] Where No. = serial number, and Rel.Int = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form A.

[0184] From Table 10 and Figure 7 It can be seen that, expressed as 2θ angle, crystal form A has the strongest characteristic absorption peak at 20.41°, with a relative absorption intensity of 100%.

[0185] Furthermore, crystal form A also has characteristic peaks at 5.00°, 7.70°, 12.82°, 14.08°, 15.44°, 16.72°, 17.17°, 19.70°, 20.88°, 23.08°, 23.78°, and 26.83°, with relative absorption intensities greater than 50%.

[0186] Furthermore, crystal form A also exhibits crystalline values ​​at 9.24°, 11.40°, 11.63°, 12.23°, 13.85°, 14.80°, 15.71°, 15.96°, 17.75°, 17.93°, 18.20°, 18.67°, 18.94°, 21.15°, 21.48°, 22.51°, 22.71°, and 22.44°. Characteristic peaks are observed at 24.65°, 25.20°, 25.66°, 25.84°, 26.60°, 27.47°, 27.74°, 27.92°, 28.81°, 29.57°, 30.27°, 30.72°, 31.38°, 32.11°, 32.70°, 33.57°, and 36.70°, with relative absorption intensities greater than 20%.

[0187] Furthermore, crystal form A also exhibits characteristic peaks at 9.88° and 13.30°, with relative absorption intensities greater than 10%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0188] Example 7: Preparation and characterization of crystal form B

[0189] 7.1 Preparation of Crystal Form B

[0190] Weigh 19.1 mg of compound 1A, add 0.9 mL of ethyl acetate / n-heptane (1 / 2, v / v), suspend and stir at low temperature (-20℃) for a certain period of time, centrifuge the suspension and dry it under vacuum at 40℃ to obtain crystal form B.

[0191] 7.2 Characterization of Crystal Form B

[0192] The crystal form B prepared in 7.1 above was subjected to XRD characterization, and the relevant characterization results are as follows: Figure 9 As shown in Table 12, the main characteristic absorption peaks of crystal form B are as follows:

[0193] Table 12 shows the XRD data for crystal form B.

[0194]

[0195] Where No. = serial number, and Rel.Int = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form B.

[0196] From Table 12 and Figure 9 It can be seen that, expressed as 2θ angle, crystal form B has the strongest characteristic absorption peak at 5.97°, with a relative absorption intensity of 100%.

[0197] Furthermore, crystal form B also has characteristic peaks at 7.80°, 9.26°, 11.57°, 14.41°, 15.17°, 17.93°, 18.12°, 19.64°, 20.10°, 22.88°, and 23.50°, with relative absorption intensities greater than 50%.

[0198] Furthermore, crystal form B also exhibits characteristic peaks at 6.73°, 10.07°, 11.12°, 12.64°, 13.44°, 14.21°, 15.63°, 15.96°, 16.74°, 17.19°, 17.42°, 18.55°, 19.29°, 20.69°, 21.52°, 22.59°, 24.59°, 25.27°, 25.66°, 26.07°, 27.22°, 28.64°, 29.41°, and 31.26°, with relative absorption intensities greater than 20%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors; for those skilled in the art, these other absorption peaks are unnecessary for characterizing this crystal form.

[0199] Example 8: Preparation and characterization of crystal form C

[0200] 8.1 Preparation of Crystal Form C

[0201] Weigh 100 mg of compound 1A, add 1 mL of isopropyl ether, suspend and stir at room temperature for a certain period of time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form C.

[0202] 8.2 Characterization of Crystal Form C

[0203] The crystalline form C obtained in step 8.1 was characterized by XRD, TGA, and DSC. The relevant characterization results are as follows: Figure 10 and Figure 11 As shown in Table 13, the main characteristic absorption peaks of crystal form C are as follows:

[0204] Table 13 shows the XRD data for crystal form C.

[0205]

[0206]

[0207] Where No. = serial number, and Rel.Int = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form C.

[0208] From Table 13 and Figure 10 It can be seen that, expressed as an angle of 2θ, crystal form C has the strongest characteristic absorption peak at 17.44°, with a relative absorption intensity of 100%.

[0209] Furthermore, crystalline form C also has characteristic peaks at 6.03°, 14.45°, 16.31°, 18.16°, 21.02°, 24.40°, 24.98°, 26.48°, and 27.84°, with relative absorption intensities greater than 50%.

[0210] Furthermore, crystalline form C also exhibits characteristic peaks at 9.28°, 10.46°, 12.37°, 12.95°, 13.61°, 15.32°, 18.86°, 22.01°, 22.77°, 23.41°, 25.60°, 26.98°, 29.08°, 30.58°, 31.71°, and 32.72°, with relative absorption intensities greater than 20%. This allows for a more detailed differentiation between other substances representing this crystalline form. Other weak absorption peaks may vary significantly due to experimental errors; for those skilled in the art, these other absorption peaks are unnecessary for characterizing this crystalline form.

[0211] Example 9: Preparation and characterization of crystal form D

[0212] 9.1 Preparation of crystal form D

[0213] Weigh 100 mg of compound 1A, add 1 mL of methyl tert-butyl ether, suspend and stir at room temperature for a certain period of time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form D.

[0214] 9.2 Characterization of crystal form D

[0215] The crystal form D prepared in section 9.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 12 and Figure 13 As shown in Table 14, the main characteristic absorption peaks of crystal form D are as follows:

[0216] Table 14 XRD data for crystal form D

[0217]

[0218] Where No. = serial number, and Rel.Int = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form D.

[0219] From Table 14 and Figure 12 It can be seen that, expressed as 2θ angle, crystal form D has the strongest characteristic absorption peak at 18.37°, with a relative absorption intensity of 100%.

[0220] Furthermore, crystal form D also has a characteristic peak at 4.15°, with a relative absorption intensity greater than 50%.

[0221] Furthermore, crystal form D also exhibits characteristic peaks at 7.12°, 9.18°, 10.46°, 12.31°, 12.62°, 13.05°, 14.31°, 15.17°, 15.52°, 15.89°, 16.86°, 19.68°, 20.74°, 21.09°, 22.14°, 24.46°, 24.90°, and 25.58°, with relative absorption intensities greater than 20%.

[0222] Furthermore, crystal form D also exhibits characteristic peaks at 13.51°, 14.70°, 16.53°, 17.29°, 26.69°, 27.45°, 28.19°, 28.91°, 30.05°, 30.58°, 30.81°, 32.43°, and 33.55°, with relative absorption intensities greater than 10%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0223] Example 10: Preparation and characterization of crystal form E

[0224] 10.1 Preparation of Crystal Form E

[0225] Weigh 100 mg of compound 1A, add 1 mL of isopropyl acetate, suspend and stir at room temperature for a certain period of time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form E.

[0226] 10.2 Characterization of Crystal Form E

[0227] The crystal form E prepared in section 10.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 14 and Figure 15 As shown in Table 15, the main characteristic absorption peaks of crystal form E are as follows:

[0228] Table 15 shows the XRD data for crystal form E.

[0229]

[0230] Where No. = serial number, and Rel.Int = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form E.

[0231] From Table 15 and Figure 14 It can be seen that, expressed as 2θ angle, crystal form E has the strongest characteristic absorption peak at 17.40°, with a relative absorption intensity of 100%.

[0232] Furthermore, crystal form E also exhibits characteristic peaks at 9.22°, 10.72°, 12.58°, 14.72°, 16.14°, 16.78°, 18.45°, 19.73°, 20.16°, 20.73°, 21.33°, 23.21°, 24.98°, 25.25°, and 27.82°, with relative absorption intensities greater than 20%.

[0233] Furthermore, crystal form E also exhibits characteristic peaks at 8.44°, 13.24°, 15.21°, 18.00°, 19.02°, 22.36°, 23.58°, 24.22°, 26.19°, 26.60°, 27.29°, 29.34°, 30.05°, 31.90°, 32.41°, 34.12°, and 40.18°, with relative absorption intensities greater than 10%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors; for those skilled in the art, these other absorption peaks are unnecessary for characterizing this crystal form.

[0234] Example 11: Preparation and characterization of crystal form F

[0235] 11.1 Preparation of Crystal Form F - Example 1

[0236] Weigh 100 mg of compound 1A, add 1 mL of dichloromethane, and suspend and stir at room temperature for a certain period of time until dissolved. Add 4 mL of n-heptane and stir to precipitate a solid. Centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form F.

[0237] The X-ray diffraction pattern of the obtained crystal form F is as follows Figure 16As shown, the specific characteristic absorption peaks are: 3.80°, 5.03°, 5.34°, 6.85°, 10.05°, 10.64°, 10.86°, 12.75°, 12.95°, 13.52°, 13.72°, 14.26°, 15.13°, 15.37°, 16.01°, 16.21°, 16.95°, and 17. Characteristic peaks are found at 35°, 18.20°, 18.44°, 19.30°, 20.17°, 21.29°, 22.01°, 22.30°, 22.73°, 23.61°, 24.82°, 25.55°, 26.05°, 26.32°, 27.21°, 28.09°, and 30.13°, with an error of ±0.2°.

[0238] The obtained crystal form F was characterized by TGA and DSC, and the relevant characterization results are as follows: Figure 17 As shown, the DSC has a characteristic absorption peak at 129.4°C, with an error of ±5°C.

[0239] 11.2 Preparation of Crystal Form F - Example 2

[0240] Weigh out X kg of crude compound 1A, start stirring, add 0.1 kg of activated carbon, protect from light, heat to 50–60 °C, stir and decolorize for about 30 min, filter while hot, and wash with ethyl acetate. Concentrate the mother liquor under reduced pressure at 40–50 °C until no obvious runoff occurs, protect from light, add 3.2 kg of ethanol, stir to dissolve, cool to 20–30 °C and stir until a large amount of solid precipitates (if no precipitation occurs after 4 h, add a small amount of seed crystals), slowly add 8.2 kg of n-heptane, stir to grow crystals for 2–3 h after addition, centrifuge, and wash with n-heptane. Dry the wet product under vacuum in water at 45–55 °C to obtain a white solid crystalline form F.

[0241] The X-ray diffraction pattern of the obtained crystal form F is as follows Figure 18 As shown, the specific characteristic absorption peaks are: 3.79°, 5.01°, 5.32°, 6.83°, 10.02°, 10.64°, 10.84°, 12.73°, 12.95°, 13.51°, 13.73°, 14.21°, 15.11°, 15.35°, 15.97°, 16.19°, 16.94°, and 17. Characteristic peaks are found at 34°, 18.12°, 18.41°, 19.27°, 20.13°, 21.27°, 21.96°, 22.25°, 22.70°, 23.60°, 24.81°, 25.51°, 26.04°, 26.28°, 27.17°, 28.00°, and 30.07°, with an error of ±0.2°.

[0242] in, Figure 16 and Figure 18The comparison of the main absorption peaks is shown in Table 16 below, with an error of ±0.2°.

[0243] Table 16 shows the XRD data for crystal form F.

[0244]

[0245] Where No. = serial number, Rel.Int. = Relative Intensity, with an error of ±0.2°. Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form F.

[0246] Summary: Based on Figure 16 and Figure 18 The XRD spectrum and XRD characteristic peak data in Table 15 are shown. The strongest characteristic absorption peak is at 17.35°, expressed as a 2θ angle, with an error of ±0.2° and a relative absorption intensity of 100%.

[0247] Furthermore, the crystal form F also has characteristic peaks at 12.75°, 15.13°, 15.37°, 16.01°, 16.21°, 18.44°, and 19.30° with an error of ±0.2° and a relative absorption intensity greater than 50%.

[0248] Furthermore, the crystal form F has characteristic peaks at 5.03°, 5.34°, 6.85°, 10.05°, 10.86°, 12.95°, 13.52°, 14.26°, 18.20°, 20.17°, 22.01°, 22.73°, 23.61°, 24.82°, 25.55°, 26.05°, 26.32°, and 28.09°, with an error of ±0.2° and a relative absorption intensity greater than 20%.

[0249] Furthermore, the described crystal form F also exhibits characteristic peaks at 3.80°, 10.64°, 13.72°, 16.95°, 21.29°, 22.30°, 27.21°, and 30.13°, with an error of ±0.2° and a relative absorption intensity greater than 10%. This allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0250] Example 12: Preparation and characterization of crystal form G

[0251] 12.1 Preparation of crystal form G

[0252] Weigh 100 mg of compound 1A, add 1 mL of ethyl acetate, and suspend and stir at room temperature for a certain period of time until dissolved. Add 4 mL of n-heptane and stir to precipitate a solid. Centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form G.

[0253] 12.2 Characterization of crystal form G

[0254] The crystal form G prepared in step 12.2 was characterized by XRD, and the relevant characterization results are as follows: Figure 19 As shown in Table 17, the main characteristic absorption peaks of crystal form G are as follows:

[0255] Table 17 shows the XRD data for crystal form G.

[0256]

[0257] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form G.

[0258] Depend on Figure 19 As shown in Table 17, crystal form G has the strongest characteristic absorption peak at 10.07°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0259] Furthermore, the crystal form G also has characteristic peaks at 5.06°, 6.46°, 9.24°, and 15.09°, with a relative absorption intensity greater than 50%.

[0260] Furthermore, the crystal form G also has characteristic peaks at 8.25°, 10.29°, 11.32°, 12.39°, 17.69°, 18.02°, 18.51°, 19.33°, 20.14°, 21.08°, 21.33°, 22.53°, 22.88°, 25.80°, and 29.34°, with a relative absorption intensity greater than 20%.

[0261] Furthermore, the described crystal form G also exhibits characteristic peaks at 12.82°, 13.61°, 14.20°, 14.62°, 15.79°, 16.29°, 16.57°, 20.59°, 21.78°, 23.21°, 23.66°, 24.24°, 24.85°, 27.70°, 30.37°, and 31.51°, with relative absorption intensities greater than 10%. These peaks allow for more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors; therefore, for those skilled in the art, these other absorption peaks are unnecessary for characterizing this crystal form.

[0262] Example 13: Preparation and characterization of crystal form H

[0263] 13.1 Characterization of crystal form H

[0264] Take 50 mg of crystal form G and place it in a vacuum drying oven. Dry it at 50°C for 1 day to obtain crystal form H.

[0265] 13.2 Characterization of crystal form H

[0266] The crystal form H obtained in section 13.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 20 and Figure 21 As shown in Table 18, the main characteristic absorption peak data for crystal form H are as follows:

[0267] Table 18 shows the XRD data for crystal form H.

[0268]

[0269] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form H.

[0270] Depend on Figure 20 As shown in Table 18, crystal form H has the strongest characteristic absorption peak at 4.69°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0271] Furthermore, the crystal form H also has characteristic peaks at 6.52°, 10.13°, 14.97°, 15.91°, 16.94°, 18.41°, 19.17°, 19.99°, 21.27°, and 22.49°, with a relative absorption intensity greater than 50%.

[0272] Furthermore, the crystal form H also exhibits characteristic peaks at 9.24°, 10.50°, 12.39°, 13.59°, and 25.66°, with relative absorption intensities greater than 20%. This allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0273] Example 14: Preparation and characterization of crystal form I

[0274] 14.1 Preparation of Crystal Form I

[0275] Weigh 100 mg of compound 1A, add 0.5 mL of cyclohexanone, suspend and stir at room temperature for a certain time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form I.

[0276] 14.2 Characterization of Crystal Form I

[0277] The crystal form I prepared in section 14.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 22 and Figure 23 As shown in Table 19, the main characteristic absorption peak data for crystal form I are as follows:

[0278] Table 19 shows the XRD data for crystal form I.

[0279]

[0280] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation of the specific crystal form I.

[0281] Depend on Figure 22 As shown in Table 19, crystal form I has the strongest characteristic absorption peak at 17.91°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0282] Furthermore, the crystal form I also has characteristic peaks at 9.47°, 13.46°, 15.07°, 19.19°, 21.33°, and 25.39°, with a relative absorption intensity greater than 50%.

[0283] Furthermore, crystal form I also exhibits characteristic peaks at 5.93°, 8.79°, 11.36°, 12.02°, 12.60°, 13.98°, 14.35°, 15.52°, 16.31°, 16.80°, 17.29°, 19.97°, 21.87°, 22.38°, 22.82°, 23.93°, 24.22°, 25.12°, 26.65°, 27.16°, 28.31°, 28.77°, 29.08°, 29.67°, 30.52°, and 31.42°, with relative absorption intensities greater than 20%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0284] Example 15: Preparation and characterization of crystal form J

[0285] 15.1 Preparation of crystal form J

[0286] Weigh 200 mg of compound 1A, add 0.5 mL of dichloromethane, and suspend and stir at room temperature for a certain period of time until dissolved. Add 2 mL of n-heptane and stir to precipitate a solid. Centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form J.

[0287] 15.2 Characterization of crystal form J

[0288] The crystal form J prepared in section 15.1 above was subjected to XRD characterization, and the relevant characterization results are as follows: Figure 24 As shown in Table 20, the main characteristic absorption peaks of crystal form J are as follows:

[0289] Table 20 shows the XRD data for crystal form J.

[0290]

[0291] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form J.

[0292] Depend on Figure 24 As shown in Table 20, crystal form J has the strongest characteristic absorption peak at 5.18°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0293] Furthermore, the crystal form J also exhibits characteristic peaks at 10.25°, 12.93°, 13.51°, 14.53°, 15.24°, 16.55°, 17.52°, 18.45°, 19.38°, 24.83°, 25.72°, and 28.01°, with relative absorption intensities greater than 50%. This allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0294] Example 16: Preparation and characterization of crystal form K

[0295] 16.1 Preparation of crystal form K

[0296] Weigh 400 mg of compound 1A, add 2 mL of anisole, suspend and stir at room temperature for a certain period of time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form K.

[0297] 16.2 Characterization of crystal form K

[0298] The crystal form K obtained in section 16.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 25 As shown in Table 21, the main characteristic absorption peaks of crystal form K are as follows:

[0299] Table 21 shows the XRD data for crystal form K.

[0300]

[0301]

[0302] Crystal form K was prepared using the preparation method described in section 16.1. Crystal form K was then used again for XRD characterization. The specific characterization results are as follows: Figure 26 As shown in Table 22, the XRD characterization results of the two crystal forms K are compared, with an error of ±0.2°.

[0303] Table 22 shows the XRD data of crystal form K after two characterizations.

[0304]

[0305]

[0306] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form K.

[0307] Depend on Figure 25 As shown in Table 21, crystal form K has the strongest characteristic absorption peak at 19.93°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0308] Furthermore, the crystal form K also has characteristic peaks at 15.28°, 17.75°, 19.54°, 20.80°, and 22.34°, with a relative absorption intensity greater than 50%.

[0309] Furthermore, the crystal form K also has characteristic peaks at 5.00°, 7.49°, 11.40°, 12.80°, 14.02°, 15.56°, 16.43°, 16.84°, 17.07°, 18.10°, 18.70°, 18.90°, 21.29°, 22.65°, 23.70°, 23.87°, 24.36°, 24.59°, 25.04°, 25.76°, 26.48°, 27.29°, 27.84°, 28.27°, 29.47°, 30.05°, and 31.30°, with a relative absorption intensity greater than 20%.

[0310] Furthermore, the crystal form K also exhibits characteristic peaks at 9.12°, 9.86°, 12.15°, 13.20°, 14.72°, 18.47°, 22.90°, 30.52°, 30.89°, 32.66°, 33.53°, 34.18°, 35.55°, 35.92°, 36.14°, 36.51°, 37.38°, 38.12°, 38.49°, 38.94°, 40.18°, 42.49°, and 42.92°, with relative absorption intensities greater than 10%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0311] Example 17: Preparation and characterization of crystal form L

[0312] 17.1 Preparation of crystal form L

[0313] Weigh 400 mg of compound 1A, add 2 mL of cyclopentanone, suspend and stir at room temperature for a certain time, then add 8 mL of methyl ether, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form L.

[0314] 17.2 Characterization of crystal form L

[0315] The crystal form L obtained in section 17.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 27 As shown in Table 23, the main characteristic absorption peak data of crystal form L are as follows:

[0316] Table 23 shows the XRD data for crystal form L.

[0317]

[0318] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form L.

[0319] Depend on Figure 27 As shown in Table 23, crystal form L has the strongest characteristic absorption peak at 16.62°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0320] Furthermore, the crystal form L also has characteristic peaks at 17.56° and 21.60°, with a relative absorption intensity greater than 50%;

[0321] Furthermore, the crystal form L also has characteristic peaks at 6.87°, 7.98°, 8.81°, 12.37°, 13.22°, 14.20°, 14.64°, 15.24°, 15.73°, 18.16°, 18.63°, 19.70°, 20.10°, 20.41°, 20.76°, 22.24°, 23.08°, 23.74°, 24.53°, 25.00°, 25.35°, 26.38°, 26.71°, 27.49°, 27.84°, 28.46°, 29.86°, and 30.83°, with a relative absorption intensity greater than 20%.

[0322] Furthermore, the crystal form L also exhibits characteristic peaks at 9.34°, 10.06°, 13.59°, 13.85°, 31.63°, 32.23°, and 33.49°, with relative absorption intensities greater than 10%. This allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0323] Example 18: Preparation and characterization of crystal form M

[0324] 18.1 Preparation of crystal form M

[0325] Weigh 400 mg of compound 1A, add 1 mL of ethyl acetate, suspend and stir at room temperature for a certain period of time, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form M.

[0326] 18.2 Characterization of crystal form M

[0327] The crystal form M prepared in section 18.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 28 and Figure 29 As shown in Table 22, the main characteristic absorption peaks of crystal form M are as follows:

[0328] Table 24 shows the XRD data for crystal form M.

[0329]

[0330] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form M.

[0331] Depend on Figure 28 As shown in Table 24, crystal form M has the strongest characteristic absorption peak at 20.08°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0332] Furthermore, the crystal form M also has characteristic peaks at 6.42°, 9.24°, 10.06°, 13.55°, 13.71°, 15.05°, 15.67°, 16.49°, 17.52°, 18.06°, 18.51°, 19.13°, 21.25°, 22.53°, 24.42°, 25.31°, and 25.58°, with relative absorption intensities greater than 50%.

[0333] Furthermore, the crystal form M also exhibits characteristic peaks at 8.19°, 11.26°, 12.19°, 12.93°, 14.47°, 17.07°, 20.88°, 21.85°, 23.43°, 24.05°, 24.75°, 26.05°, 26.48°, 27.24°, 27.94°, 29.04°, 29.80°, 30.23°, 30.99°, 31.32°, 32.06°, and 33.32°, with relative absorption intensities greater than 20%. This allows for a more detailed differentiation between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0334] Example 19: Preparation and characterization of crystalline N

[0335] 19.1 Preparation of Crystal Form N

[0336] Weigh 400 mg of compound 1A, add 1 mL of cyclic ketone, suspend and stir at room temperature for a certain time, then add 4 mL of methyl ether, centrifuge the suspension and dry it under vacuum at 40 °C to obtain crystal form N.

[0337] 19.2 Characterization of N crystal form

[0338] The N crystal form prepared in section 19.1 above was characterized by XRD, TGA and DSC. The relevant characterization results are as follows: Figure 30 and Figure 31 As shown in the table. The main characteristic absorption peaks of crystalline N are shown in Table 25:

[0339] Table 25 shows the XRD data for crystal form N.

[0340]

[0341] Crystal form N was prepared using the method described in section 19.1. Crystal form N was then used again for XRD characterization. The specific characterization results are as follows: Figure 32 As shown in the figure. The comparison of the XRD characterization results of the two crystal forms N is shown in Table 26, with an error of ±0.2°.

[0342] Table 26 shows the XRD data of crystal form N after two characterizations.

[0343]

[0344] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form N.

[0345] Depend on Figure 30 As shown in Table 25, crystal form N has the strongest characteristic absorption peak at 16.49°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0346] Furthermore, the N crystal form also has a characteristic peak at 17.46°, with a relative absorption intensity greater than 50%;

[0347] Furthermore, the crystalline form N also has characteristic peaks at 7.88°, 8.71°, 13.13°, 18.63°, 19.58°, 19.93°, 21.48°, and 22.96°, with a relative absorption intensity greater than 20%.

[0348] Furthermore, the crystal form N also exhibits characteristic peaks at 11.45°, 14.10°, 15.19°, 15.34°, 17.97°, 20.78°, 22.55°, 23.27°, 23.91°, 24.42°, 24.96°, 25.29°, 26.32°, 27.18°, 28.44°, 28.81°, 29.12°, 29.47°, 30.72°, and 34.27°, with relative absorption intensities greater than 10%. This allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may vary significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0349] Example 20: Preparation and characterization of crystal form O

[0350] 20.1 Preparation of Crystal Form O

[0351] Crystalline O can be obtained by suspending a wet sample of a highly concentrated amorphous raw material in ethanol at room temperature.

[0352] Weigh 100 mg of compound 1A, add 1 mL of ethanol, suspend and stir at room temperature for a certain period of time, and then centrifuge the suspension to obtain crystal form O.

[0353] 20.2 Characterization of crystal form O

[0354] The crystal form O prepared in step 20.1 above was characterized by XRD, and the relevant characterization results are as follows: Figure 33 As shown in Table 27, the main characteristic absorption peak data of crystal form O are as follows:

[0355] Table 27 shows the XRD data for crystal form O.

[0356]

[0357] Where No. = serial number, and Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form O.

[0358] Depend on Figure 33 As shown in Table 27, crystal form O has the strongest characteristic absorption peak at 14.68°, with a relative absorption intensity of 100%, expressed as 2θ angle.

[0359] Furthermore, the crystal form O also has characteristic peaks at 5.22°, 16.66°, 18.61°, 19.15°, 19.83°, and 23.41°, with a relative absorption intensity greater than 50%.

[0360] Furthermore, the crystalline form O also exhibits characteristic peaks at 6.54°, 8.31°, 9.32°, 9.59°, 10.37°, 13.03°, 15.26°, 15.54°, 17.64°, 18.02°, 18.74°, 19.58°, 20.51°, 21.23°, 21.85°, 23.74°, 24.28°, 24.57°, 25.04°, 26.11°, 27.08°, 28.11°, 28.40°, 28.83°, 29.53°, and 31.28°, with relative absorption intensities greater than 20%.

[0361] Furthermore, the crystal form O also has characteristic peaks at 33.13° and 33.63°, with relative absorption intensities greater than 10%; this allows for a more detailed distinction between other substances representing this crystal form. Other weak absorption peaks may change significantly due to experimental errors, and for those skilled in the art, these other absorption peaks are considered unnecessary for characterizing this crystal form.

[0362] Example 21: Stability Study

[0363] The stability of crystal forms A, F, K, N, and amorphous samples under illumination (25℃ / 4500 Lux) was studied. Sampling times were 0 days and 26 days, and the HPLC purity of the samples was examined. The results are shown in Table 28.

[0364] Table 28 shows the stability study results for crystalline forms A, F, K, N, and amorphous forms.

[0365]

[0366]

[0367] As shown in Table 28, crystal forms A, F, K, and N have good stability under light conditions, and crystal forms A, F, K, and N have higher purity and better stability than the amorphous samples.

[0368] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A crystalline form A of a terpyridinedione compound of formula (I), characterized in that: The X-ray diffraction pattern of crystal form A shows characteristic peaks at 5.00°, 7.70°, 9.24°, 11.40°, 12.23°, 12.82°, 14.08°, 14.80°, 15.44°, 15.71°, 16.72°, 17.17°, 17.75°, 18.67°, 18.94°, 19.70°, 20.41°, 20.88°, 21.15°, 21.48°, 22.71°, 23.78°, 24.44°, 25.20°, 25.66°, 25.84°, 26.60°, 27.47°, 28.81°, 29.57°, and 30.27°, with an error of ±0.2°.

2. The crystalline form A of the terpyridinedione compound of formula (I) according to claim 1, characterized in that, The X-ray diffraction pattern of crystal form A is basically shown in Figure 7 or Figure 8.

3. A crystalline form F of a terpyridinedione compound represented by formula (I), characterized in that: The X-ray diffraction patterns of crystal form F are represented by 2θ angles at 3.80°, 5.03°, 5.34°, 6.85°, 10.05°, 10.64°, 10.86°, 12.75°, 12.95°, 13.52°, 13.72°, 14.26°, 15.13°, 15.37°, 16.01°, 16.21°, and 16.95°. Characteristic peaks are observed at 17.35°, 18.20°, 18.44°, 19.30°, 20.17°, 21.29°, 22.01°, 22.30°, 22.73°, 23.61°, 24.82°, 25.55°, 26.05°, 26.32°, 27.21°, 28.09°, and 30.13°, with an error of ±0.2°.

4. The crystalline form F of the terpyridinedione compound of formula (I) according to claim 3, characterized in that, The X-ray diffraction pattern of the crystal form F is basically shown in Figure 16 or Figure 18.

5. The crystalline form F of the terpyridinedione compound of formula (I) according to claim 3, characterized in that: The differential thermal analysis spectrum of the crystal form F shows a characteristic absorption peak at 129.4℃, with an error of ±5℃.

6. A crystalline form K of a terpyridinedione compound represented by formula (I), characterized in that: The X-ray diffraction pattern of crystal form K is represented by a 2θ angle at 5.00°, 7.49°, 9.12°, 11.40°, 12.80°, 14.02°, 14.72°, 15.28°, 15.56°, 16.43°, 16.84°, 17.07°, 17.75°, 18.10°, 18.47°, 18.70°, and 18.90°. Characteristic peaks are observed at 19.54°, 19.93°, 20.80°, 21.29°, 22.34°, 22.65°, 23.70°, 24.36°, 24.59°, 25.04°, 25.76°, 26.48°, 27.29°, 27.84°, 28.27°, 29.47°, and 30.05°, with an error of ±0.2°.

7. The crystalline form K of the terpyridinedione compound of formula (I) according to claim 6, characterized in that, The X-ray diffraction pattern of crystal form K is basically shown in Figure 25 or Figure 26.

8. A crystalline form N of a terpyridinedione compound of formula (I), characterized in that: The X-ray diffraction pattern of the N crystal form shows characteristic peaks at 7.88°, 8.71°, 13.13°, 14.10°, 15.34°, 16.49°, 17.46°, 18.63°, 19.58°, 19.93°, 21.48°, 22.96°, 25.29°, and 26.32°, with an error of ±0.2°.

9. The crystalline form N of the terpyridinedione compound of formula (I) according to claim 8, characterized in that, The X-ray diffraction pattern of the N crystal form is basically shown in Figure 30 or Figure 32.

10. A pharmaceutical composition, characterized in that, The crystal form and pharmaceutically acceptable carrier comprising a therapeutically effective amount of any one of claims 1-9.

11. Use of the crystal form according to any one of claims 1-9 in the preparation of a medicament for treating a disease, said disease being a p38 / MK2-related disease selected from chronic inflammatory diseases and acute inflammatory diseases.

12. The use according to claim 11, wherein the chronic inflammatory condition is rheumatoid arthritis.

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