Polymorphs of IRAK-4 degrading agent

Abstract

Disclosed in the present invention are polymorphs of an IRAK-4 degrading agent. Specifically, the present invention provides a plurality of crystal forms of an IRAK-4 degrading agent represented by the following formula (I), salts thereof and crystal forms thereof, as well as a pharmaceutical composition thereof, the preparation therefor and the use thereof. The crystal forms provided by the present invention can be used for preparing drugs for treating and / or preventing IRAK4-mediated related diseases.

Description

A polymorph of an IRAK-4 degrader

[0001] This application claims priority to Chinese Patent Application No. 2024111779751, filed August 26, 2024, and Chinese Patent Application No. 202511166071.3, filed August 19, 2025. The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of medicinal chemistry, specifically relating to multiple crystal forms of an IRAK-4 degrader, its salts and crystal forms thereof, its pharmaceutical compositions, preparation and uses. Background Technology

[0003] Proteolysis-targeting chimaeras (PROTACs) technology is an emerging drug development technology that has emerged in recent years. Unlike the mechanism of action of traditional small-molecule inhibitors, PROTACs work by delivering these proteins into the proteasome for complete degradation, thus breaking the traditional definition of drugs. Compared with traditional small-molecule inhibitors, they have several advantages, such as making the target "drugable" instead of "undruggable," not relying on "site-driven" mechanisms, and overcoming the problem of drug resistance.

[0004] Interleukin-1 receptor-associated kinase 4 (IRAK-4) is a member of the intracellular serine-threonine kinase family, the IRAK family. It mediates Toll-like receptor (TLR) and IL-1 receptor (IL-1R) activation-mediated inflammatory signaling pathways, playing a crucial role in innate immune signaling. IRAK4 can be activated by recruitment by MyD88 to form Myddosomes, thereby activating downstream IRAK1 / 2, leading to subsequent hyperphosphorylation, dissociation from the complex, and binding to TRAF6. The activated TRAF6 complex triggers downstream signaling pathways to produce pro-inflammatory cytokines (NF-κB, CREB, AP-1, IRF, etc.). Inhibition of IRAK4 may play a significant role in the pathogenesis and progression of inflammatory diseases. Currently, inhibitors targeting IRAK4 are not approved. Therefore, knocking out IRAK4 using PROTAC technology may be another strategy for treating IRAK4-related diseases.

[0005] PCT / CN2024 / 079646 describes an IRAK4 degrader with the structure shown in formula (I) below. Its chemical name is 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-(7-(4-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)but-3-yn-1-yl)-7-azaspiro[3.5]nonane-2-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-amide. This compound exhibits good in vitro and in vivo efficacy and is expected to be developed into a clinical drug.

[0006] In addition, the study of crystal forms plays an important role in the drug development process. Different crystal forms of the same drug have significant differences in physicochemical and biological properties. In order to better control the quality of drugs and meet the requirements of formulation, production, storage and transportation, we studied the salts and crystal forms of compounds of formula (I) in order to discover salts or crystal forms with good properties. Summary of the Invention

[0007] This disclosure relates to various crystal forms of the compound 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-(7-(4-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)but-3-yn-1-yl)-7-azaspiro[3.5]nonane-2-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-amide, methods for their preparation, and their use in the preparation of medicaments for the treatment and / or prevention of IRAK4-mediated related diseases.

[0008] All contents contained in PCT / CN2024 / 079646 by the applicant are incorporated herein by reference.

[0009] In a first aspect, this disclosure provides a crystal form A of a compound of formula (I),

[0010] Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.48, 10.34, 10.98 and 14.19, with a 2θ error range of ±0.2°.

[0011] In some embodiments of this disclosure, the crystal form A, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 5.31, 9.22, 10.76, 16.17, 18.30, 22.15, and 25.36, with a 2θ error range of ±0.2°.

[0012] In some embodiments of this disclosure, the crystal form A, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 11.76, 12.67, 13.32, 14.63, 15.11, 15.76, 16.89, 17.82, 18.76, 19.75, 20.52, 21.35, 22.87, 23.45, and 26.50, with a 2θ error range of ±0.2°.

[0013] In some embodiments of this disclosure, the crystal form A, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is essentially as shown in Figure 1.

[0014] In some embodiments of this disclosure, the crystal form A has an endothermic peak in its DSC spectrum between 180°C and 220°C; preferably, it has an endothermic peak at 204.8°C ± 2°C.

[0015] In some embodiments of this disclosure, the crystal form A has a TGA spectrum showing a weight loss of 0.7% between 35 and 180 °C.

[0016] In some embodiments of this disclosure, the TGA-DSC spectrum of crystal form A is basically as shown in Figure 2.

[0017] In some embodiments of this disclosure, the DVS spectrum of crystal form A is basically as shown in Figure 3.

[0018] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of crystal form A are shown in Table 1a below:

[0019] Table 1a shows the XRPD diffraction peak analysis data for crystal form A of compound (I).

[0020] This disclosure provides crystal form B of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.87, 10.82 and 11.52, with a 2θ error range of ±0.2°.

[0021] In some embodiments of this disclosure, the crystal form B, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 13.58, 16.45, 17.00, 18.86, 19.76, 20.43, 22.64, 23.76, and 27.88, with a 2θ error range of ±0.2°.

[0022] In some embodiments of this disclosure, the crystal form B, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is essentially as shown in Figure 4.

[0023] In some embodiments of this disclosure, the crystal form B has an endothermic peak in its DSC spectrum between 170°C and 210°C; preferably, it has an endothermic peak at 196.6°C ± 2°C.

[0024] In some embodiments of this disclosure, the crystal form B has a TGA spectrum showing a weight loss of 3.8% at 35–209 °C.

[0025] In some embodiments of this disclosure, the crystal form B is a methanol solvate.

[0026] In some embodiments of this disclosure, the crystal form B is a monomethanol solvate.

[0027] In some embodiments of this disclosure, the TGA-DSC spectrum of crystal form B is basically as shown in Figure 5.

[0028] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of crystal form B are shown in Table 2a below:

[0029] Table 2a shows the XRPD diffraction peak analysis data for crystal form B of compound (I).

[0030] This disclosure provides crystal form C of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.85, 8.80, 10.82, 11.46 and 13.54, with a 2θ error range of ±0.2°.

[0031] In some embodiments of this disclosure, the crystal form C, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 9.72, 12.80, 16.50, 16.87, 17.67, 18.87, 19.28, 19.80, 20.56, 22.65, 23.84, and 27.95, with a 2θ error range of ±0.2°.

[0032] In some embodiments of this disclosure, the crystal form C, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 22.10, 23.21, and 24.36, with a 2θ error range of ±0.2°.

[0033] In some embodiments of this disclosure, the crystal form C, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is essentially as shown in Figure 6.

[0034] In some embodiments of this disclosure, the DSC spectrum of the crystal form C shows an endothermic peak between 170°C and 220°C; preferably, it has an endothermic peak at 195.7°C ± 2°C.

[0035] In some embodiments of this disclosure, the crystal form C has a TGA spectrum showing a weight loss of 5.7% at 36–213 °C.

[0036] In some embodiments of this disclosure, the crystal form C is an ethanol solvate.

[0037] In some embodiments of this disclosure, the crystal form C is a monoethanol solvate.

[0038] In some embodiments of this disclosure, the TGA-DSC spectrum of the crystal form C is basically as shown in Figure 7.

[0039] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of crystal form C are shown in Table 3a below:

[0040] Table 3a shows the XRPD diffraction peak analysis data for crystal form C of compound (I).

[0041] This disclosure provides crystal form D of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.81, 8.81, 9.76, 10.89, 11.37 and 13.60, with a 2θ error range of ±0.2°.

[0042] In some embodiments of this disclosure, the crystal form D, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 3.92, 12.78, 16.60, 16.89, 17.67, 18.78, 19.26, 19.84, 20.41, 20.69, 22.06, 22.63, 23.91, and 28.10, with a 2θ error range of ±0.2°.

[0043] In some embodiments of this disclosure, the crystal form D, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is essentially as shown in Figure 8.

[0044] In some embodiments of this disclosure, the crystal form D has an endothermic peak in its DSC spectrum between 170°C and 220°C; preferably, it has an endothermic peak at 202.35°C ± 2°C.

[0045] In some embodiments of this disclosure, the crystal form D has a TGA spectrum showing a weight loss of 7.1% at 36–217 °C.

[0046] In some embodiments of this disclosure, the crystal form D is an isopropanol solvate.

[0047] In some embodiments of this disclosure, the crystal form D is a monoisopropanol solvate.

[0048] In some embodiments of this disclosure, the TGA-DSC spectrum of the crystal form D is basically as shown in Figure 9.

[0049] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of the crystal form D are shown in Table 4a below:

[0050] Table 4a shows the XRPD diffraction peak analysis data for crystal form D of compound (I).

[0051] This disclosure provides crystal form E of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.78, 9.74, 10.76, 11.48 and 13.46, with a 2θ error range of ±0.2°.

[0052] In some embodiments of this disclosure, the crystal form E, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 15.15, 16.54, 16.84, 17.56, 18.82, 19.28, 19.80, 20.30, 22.60, 23.80, and 27.80, with a 2θ error range of ±0.2°.

[0053] In some embodiments of this disclosure, the crystal form E is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is essentially as shown in Figure 10.

[0054] In some embodiments of this disclosure, the crystal form E has an endothermic peak in its DSC spectrum between 170°C and 220°C; preferably, it has an endothermic peak at 197.72°C ± 2°C.

[0055] In some embodiments of this disclosure, the crystal form E has a TGA spectrum showing a weight loss of 5.2% at 95–214 °C.

[0056] In some embodiments of this disclosure, the crystal form E is an acetone solvate.

[0057] In some embodiments of this disclosure, the crystal form E is a monoacetone solvate.

[0058] In some embodiments of this disclosure, the TGA-DSC spectrum of the crystal form E is basically as shown in Figure 11.

[0059] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of the crystal form E are shown in Table 5a below:

[0060] Table 5a shows the XRPD diffraction peak analysis data for crystal form E of compound E.

[0061] This disclosure provides crystal form F of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.93, 8.93, 11.04, 11.59 and 13.82, with a 2θ error range of ±0.2°.

[0062] In some embodiments of this disclosure, the crystal form F, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 4.00, 9.67, 12.87, 16.37, 17.02, 19.82, 20.71, 21.06, 24.17, and 28.34, with a 2θ error range of ±0.2°.

[0063] In some embodiments of this disclosure, the crystal form F, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 18.04, 18.52, 19.04, 19.45, 22.39, and 22.93, with a 2θ error range of ±0.2°.

[0064] In some embodiments of this disclosure, the crystal form F is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is basically as shown in Figure 12.

[0065] In some embodiments of this disclosure, the crystal form F has an endothermic peak in its DSC spectrum between 170°C and 220°C; preferably, it has an endothermic peak at 203.24°C ± 2°C.

[0066] In some embodiments of this disclosure, the crystal form F has a TGA spectrum showing a weight loss of 4.4% at 36–215 °C.

[0067] In some embodiments of this disclosure, the crystal form F is an acetonitrile solvate.

[0068] In some embodiments of this disclosure, the crystal form F is a monoacetonitrile solvate.

[0069] In some embodiments of this disclosure, the TGA-DSC spectrum of the crystal form F is basically as shown in Figure 13.

[0070] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of the crystal form F are shown in Table 6a below:

[0071] Table 6a shows the XRPD diffraction peak analysis data for crystal form F of compound (I).

[0072] This disclosure provides crystal form G of the compound of formula (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.11, 6.54, 7.91, 8.93, 9.37 and 11.06, with a 2θ error range of ±0.2°.

[0073] In some embodiments of this disclosure, the crystal form G, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 4.00, 11.56, 13.82, 16.41, 17.04, 18.02, 18.91, and 19.84, with a 2θ error range of ±0.2°.

[0074] In some embodiments of this disclosure, the crystal form G, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 12.91, 15.02, 16.04, 20.71, 20.98, 22.84, 24.19, 25.71, and 28.30, with a 2θ error range of ±0.2°.

[0075] In some embodiments of this disclosure, the crystal form G is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is essentially as shown in Figure 14.

[0076] In some embodiments of this disclosure, the crystal form G has an endothermic peak at 83.9℃±2℃ and 199.1℃±2℃ as shown in its DSC spectrum.

[0077] In some embodiments of this disclosure, the crystal form G has a TGA spectrum showing a weight loss of 2.7% at 35–118°C and a weight loss of 2.6% at 118–221°C.

[0078] In some embodiments of this disclosure, the crystal form G is an acetonitrile solvate.

[0079] In some embodiments of this disclosure, the crystal form G is a semiacetonitrile solvate.

[0080] In some embodiments of this disclosure, the TGA-DSC spectrum of the crystal form G is basically as shown in Figure 15.

[0081] In some embodiments of this disclosure, the X-ray powder diffraction pattern data of the crystal form G are shown in Table 7a below:

[0082] Table 7a shows the XRPD diffraction peak analysis data for crystal form G of compound G.

[0083] Secondly, the present invention provides pharmaceutically acceptable salts formed by a compound of formula (I) and one or more acid molecules.

[0084] The acid is selected from organic acids or inorganic acids.

[0085] In some embodiments of this disclosure, the pharmaceutically acceptable salt of the compound of formula (I) is selected from hydrochloride, maleate, p-toluenesulfonate, methanesulfonate, hydrobromide, succinate, L-malate, L-tartrate, benzenesulfonate, phosphate, gentianate, oxalate, fumarate, and citrate.

[0086] In some embodiments of this disclosure, the pharmaceutically acceptable salt of the compound of formula (I) is p-toluenesulfonate.

[0087] In some embodiments of this disclosure, the pharmaceutically acceptable salt of the compound of formula (I) is a mesylate.

[0088] Preferably, the pharmaceutically acceptable salt of the compound of formula (I) is a toluenesulfonate or a methanesulfonate.

[0089] This disclosure provides a compound of formula (I) p-toluenesulfonate crystal form I, using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 5.37, 7.41, 8.76, 9.91 and 14.24, with a 2θ error range of ±0.2°.

[0090] In some embodiments of this disclosure, the p-toluenesulfonate crystal form I of the compound of formula (I), when subjected to Cu-Kα radiation, has a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 10.86, 11.74, 14.89, 15.21, 16.43, 18.28, 21.39, and 21.67, with a 2θ error range of ±0.2°.

[0091] In certain embodiments of this disclosure, the X-ray powder diffraction pattern of compound (I) p-toluenesulfonate crystal form I, when subjected to Cu-Kα irradiation, also exhibits characteristic peaks at one or more of the following 2θ values: 12.61, 12.91, 19.43, 20.37, 22.54, 23.26, 24.21, 24.69, 25.43, 25.93, 26.84, 27.73, and 29.17, with a 2θ error range of ±0.2°.

[0092] In some embodiments of this disclosure, the X-ray powder diffraction pattern of the compound of formula (I) p-toluenesulfonate, crystal form I, using Cu-Kα radiation, is essentially as shown in Figure 16.

[0093] In some embodiments of this disclosure, the p-toluenesulfonate crystal form I of the compound of formula (I) has an endothermic peak at 50℃-130℃, preferably at 95.2℃±2℃, as shown in its DSC spectrum; and an endothermic peak at 220℃-250℃, preferably at 237.7℃±2℃.

[0094] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form I has a TGA spectrum showing a weight loss of 2.1% at 35–134 °C.

[0095] In some embodiments of this disclosure, the TGA-DSC spectrum of the compound of formula (I) p-toluenesulfonate is essentially as shown in Figure 17.

[0096] In some embodiments of this disclosure, the DVS spectrum of the compound of formula (I) p-toluenesulfonate is essentially as shown in Figure 18.

[0097] In some embodiments of this disclosure, the compound of formula (I) is p-toluenesulfonate crystal form I, and the molar ratio of the compound of formula (I) to p-toluenesulfonic acid is 1:1.

[0098] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form I is a hydrate.

[0099] In some embodiments of this disclosure, the p-toluenesulfonate crystal form I of the compound of formula (I) has a water content of no more than 6%, preferably no more than 5 ± 0.5%, preferably 1%-5%, preferably 1.5%-3.5%, preferably no more than 2.5%, and preferably no more than 2%.

[0100] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form I is a monohydrate.

[0101] In some embodiments of this disclosure, the water content in the p-toluenesulfonate crystal form I hydrate of the compound of formula (I) does not affect its crystal form.

[0102] In some embodiments of this disclosure, the single-crystal structure data of p-toluenesulfonate I of the compound of formula (I) are as follows:

[0103] The space group is P21, and the cell parameters are... α=90°, β=91.1(4)°, γ=90°, The smallest asymmetric unit contains two molecules of compound (I), two p-toluenesulfonic acid molecules, and two water molecules, and each unit cell contains two smallest asymmetric units.

[0104] In some embodiments of this disclosure, the two compound molecules of formula (I) in the single crystal structure of p-toluenesulfonate I are enantiomers.

[0105] In some embodiments of this disclosure, the X-ray powder diffraction data of the p-toluenesulfonate crystal form I of compound (I) are shown in Table 8a below:

[0106] Table 8a shows the XRPD diffraction peak analysis data for p-toluenesulfonate crystal form I of compound (I).

[0107] In some embodiments of this disclosure, the crystallographic parameters of the single-crystal diffraction structure of p-toluenesulfonate of formula (I) are shown in Table 8b below:

[0108] Table 8b shows the single-crystal structure information of compound (I) p-toluenesulfonate crystal form I.

[0109] This disclosure provides a p-toluenesulfonate crystal form II of compound (I), using Cu-K αThe radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.35, 8.74, 9.84, 11.69, 14.65 and 18.19, with a 2θ error range of ±0.2°.

[0110] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form II is used with Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 12.54, 16.43 and 20.60, with a 2θ error range of ±0.2°.

[0111] In some embodiments of this disclosure, the X-ray powder diffraction pattern of the compound of formula (I) p-toluenesulfonate II, when subjected to Cu-Kα radiation, is essentially as shown in Figure 22.

[0112] In some embodiments of this disclosure, the p-toluenesulfonate crystal form II of compound (I) has a DSC spectrum showing an endothermic peak at 70℃-130℃, preferably at 100.14℃±2℃; and an endothermic peak at 220℃-250℃, preferably at 223.2℃±2℃.

[0113] In some embodiments of this disclosure, the p-toluenesulfonate crystal form II of compound (I) shows a weight loss of 2.3% at 36–169 °C according to its TGA spectrum.

[0114] In some embodiments of this disclosure, the TGA-DSC spectrum of the compound of formula (I) p-toluenesulfonate crystal form II is basically as shown in Figure 23.

[0115] In some embodiments of this disclosure, the compound of formula (I) is p-toluenesulfonate crystal form II, and the molar ratio of the compound of formula (I) to p-toluenesulfonic acid is 1:1.

[0116] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form II is a hydrate.

[0117] In some embodiments of this disclosure, the p-toluenesulfonate crystal form II of compound (I) has a water content of 1%-5%; preferably, the water content is 1.5%-3.5%; preferably, the water content does not exceed 2.5%; preferably, the water content does not exceed 2%.

[0118] In some embodiments of this disclosure, the compound of formula (I) p-toluenesulfonate crystal form II is a monohydrate.

[0119] In some embodiments of this disclosure, the X-ray powder diffraction data of p-toluenesulfonate crystal form II of compound (I) are shown in Table 9a below:

[0120] Table 9a shows the XRPD diffraction peak analysis data for compound p-toluenesulfonate crystal form II of formula (I).

[0121] This disclosure provides a methanesulfonate crystal form III of compound (I), using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.87, 9.41, 11.80, 12.13, 13.65, 15.33 and 15.86, with a 2θ error range of ±0.2°.

[0122] In some embodiments of this disclosure, the methanesulfonate form III of the compound of formula (I) is used with Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 8.72, 9.15, 17.06, 17.71, 18.04, 19.26, 22.49 and 23.69, with a 2θ error range of ±0.2°.

[0123] In some embodiments of this disclosure, the methanesulfonate form III of the compound of formula (I) is used with Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 12.67, 18.39, 19.78, 20.21, 21.02, 24.54, 25.76 and 26.50, with a 2θ error range of ±0.2°.

[0124] In some embodiments of this disclosure, the methanesulfonate crystal form III of compound (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is essentially as shown in Figure 24.

[0125] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) has an endothermic peak at 70°C-130°C, preferably at 96.77°C±2°C, as shown in its DSC spectrum.

[0126] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) shows a weight loss of 1.84% at 36–145 °C according to its TGA spectrum.

[0127] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) has a TGA-DSC spectrum that is essentially shown in Figure 25.

[0128] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound (I) is described, and the molar ratio of the compound (I) to p-toluenesulfonic acid is 1:1.

[0129] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) is a hydrate.

[0130] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) has a water content of 1%-3%; preferably, the water content is 1.5%-2.5%; preferably, the water content is no more than 2.0%.

[0131] In some embodiments of this disclosure, the methanesulfonate crystal form III of the compound of formula (I) is a monohydrate.

[0132] In some embodiments of this disclosure, the X-ray powder diffraction data of the methanesulfonate crystal form III of compound (I) are shown in Table 10a below:

[0133] Table 10a XRPD diffraction peak analysis data of methanesulfonate crystal form III of compound (I)

[0134] Thirdly, this disclosure provides a pharmaceutical formulation containing one or more of the following: a compound of formula (I) AG selected from the first aspect, a salt of the compound of formula (I) described in the second aspect, and crystal forms I-III. The formulation further contains one or more pharmaceutically acceptable excipients, and the pharmaceutical formulation may be in any pharmaceutically acceptable dosage form. Pharmaceutically acceptable excipients are substances that are non-toxic, compatible with the active ingredient, and otherwise biologically suitable for use in organisms. The selection of a specific excipient will depend on the route of administration or the type and state of disease for treating a particular patient. Examples of pharmaceutically acceptable excipients include, but are not limited to, solvents, diluents, dispersants, suspending agents, surfactants, isotonic agents, thickeners, emulsifiers, binders, lubricants, stabilizers, hydrating agents, emulsification accelerators, buffers, absorbents, colorants, ion exchangers, release agents, coating agents, flavoring agents, and antioxidants, etc., which are conventional in the pharmaceutical field. If necessary, flavoring agents, preservatives, and sweeteners may also be added to the pharmaceutical composition.

[0135] Fourthly, this disclosure provides the use of the crystal form AG of the compound of formula (I) described in the first aspect, the salt of the compound of formula (I) described in the second aspect, and any one of crystal forms I-III in the preparation of medicaments for treating and / or preventing related diseases mediated by IRAK4.

[0136] In some embodiments of this disclosure, the IRAK4-mediated diseases are selected from immune inflammatory diseases.

[0137] In some embodiments of this disclosure, the IRAK4-mediated related diseases are selected from hidradenitis suppurativa, rheumatoid arthritis, atopic dermatitis, lupus erythematosus, gouty arthritis, psoriasis, asthma, chronic obstructive pulmonary disease, polyposis sinusitis, inflammatory bowel disease, respiratory distress syndrome, chronic kidney disease, acute lung injury, IgA nephropathy, gout, multiple sclerosis, and ischemic stroke.

[0138] Fifthly, this disclosure provides a method for treating a disease, comprising administering to a subject an effective therapeutic amount of a compound of formula (I) AG as described in the first aspect, a salt of the compound of formula (I) as described in the second aspect, any one or more crystal forms of crystal forms I-III, or a pharmaceutical preparation thereof, wherein the disease is an IRAK4-mediated disease.

[0139] In some embodiments of this disclosure, the IRAK4-mediated diseases are selected from immune inflammatory diseases.

[0140] In some embodiments of this disclosure, the IRAK4-mediated related diseases are selected from hidradenitis suppurativa, rheumatoid arthritis, atopic dermatitis, lupus erythematosus, gouty arthritis, psoriasis, asthma, chronic obstructive pulmonary disease, polyposis sinusitis, inflammatory bowel disease, respiratory distress syndrome, chronic kidney disease, acute lung injury, IgA nephropathy, gout, multiple sclerosis, and ischemic stroke.

[0141] [Revised according to Rule 91, January 2026] In a sixth aspect, this disclosure provides a method for preparing p-toluenesulfonate crystal form I of compound (I), comprising the following steps:

[0142] The compound of formula (I) and a certain amount of p-toluenesulfonic acid were mixed in a solvent, stirred and slurried at room temperature for a certain period of time, and the solid was collected and dried to obtain p-toluenesulfonate crystal form I of compound (I).

[0143] [Correction 20.01.2026 according to Rule 91] Preferably, the solvent is selected from organic solvents or mixtures of organic solvents and water, wherein the organic solvent is selected from alcohols, nitrile solvents, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile solvent is selected from acetonitrile; and the ketone solvent is selected from acetone.

[0144] Preferably, the stirring time is selected from 1 hour to 96 hours;

[0145] Preferably, the molar ratio of p-toluenesulfonic acid to the compound of formula (I) is 1.0 to 1.5:1, more preferably 1.0 to 1.2:1.

[0146] [Revised according to Rule 91, January 2026] In a seventh aspect, this disclosure provides a method for preparing p-toluenesulfonate crystal form I of compound (I), comprising the following steps:

[0147] (1) The compound of formula (I) and a certain amount of p-toluenesulfonic acid are mixed in a mixed solvent of organic solvent and water and heated to a certain temperature to dissolve;

[0148] (2) After cooling to saturation, a certain amount of compound (I) p-toluenesulfonate crystal form I can be selectively added as a seed crystal to initiate crystallization;

[0149] (3) Continue cooling or optionally add a certain amount of anti-solvent to allow the product to crystallize further;

[0150] (4) Solid separation: The solid is dried to obtain p-toluenesulfonate crystal form I of compound (I).

[0151] In some embodiments of this disclosure, the molar ratio of p-toluenesulfonic acid to the compound of formula (I) is 1.0 to 1.5:1, preferably 1.0 to 1.2:1.

[0152] In some embodiments of this disclosure, cooling to solution saturation is preferably done by cooling to about 40-55°C.

[0153] In some embodiments of this disclosure, the temperature is preferably lowered to not less than 0°C.

[0154] In some embodiments of this disclosure, the organic solvent refers to an organic solvent miscible with water selected from alcohols, nitrile solvents, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile solvent is selected from acetonitrile; and the ketone solvent is selected from acetone.

[0155] In some embodiments of this disclosure, the antisolvent is selected from alcohols, nitriles, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile is selected from acetonitrile; and the ketone is selected from acetone.

[0156] In some embodiments of this disclosure, the amount of seed crystal added may be 0.1 wt% to 10 wt% of the amount of the compound of formula (I), preferably 0.5 wt% to 5 wt%.

[0157] In some embodiments of this disclosure, the method for preparing the seed crystal is as described in the sixth aspect of this disclosure.

[0158] Explanation and Definition

[0159] In this application, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of the invention, definitions of some terms are provided below. When the definitions of terms provided in this application differ from the meanings commonly understood by those skilled in the art, the definitions and interpretations provided in this application shall prevail.

[0160] Unless otherwise stated, in this disclosure, the structure of the compound is... This indicates that the atom connecting the bond is a chiral atom, and the compound can be a racemic mixture, an isomer, or a mixture of isomers with the same or different amounts.

[0161] [Revised according to Article 91, 2001.2026] The term "pharmaceutical salt" refers to the salt of the compounds of the present invention, prepared by reacting a compound having specific substituents discovered in the present invention with a relatively non-toxic acid or base. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in their free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0162] The term "effective therapeutic dose" refers to a sufficient amount of the compound of the present invention or a pharmaceutically acceptable salt thereof to treat any medical condition with a reasonable benefit / risk ratio suitable for any medical treatment and / or prevention, in order to treat the disorder. However, it should be understood that the total daily dose of the compound of formula (I) of the present invention or a pharmaceutically acceptable salt thereof and the composition thereof must be determined by the attending physician within the bounds of reliable medical judgment. For any given patient, the specific effective therapeutic dose level must be determined based on a number of factors, including the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors known in the medical field.

[0163] As is known in the art, X-ray powder diffraction patterns have one or more measurement errors due to minute variations in measurement conditions. The structures of the crystals, crystalline forms, or crystal types disclosed or claimed in this invention may exhibit similar but not identical analytical characteristics within a reasonable error range, depending on experimental conditions, purity, equipment, and other constant variables known to those skilled in the art. For example, the diffraction angle (2θ) in powder X-ray powder diffraction typically produces an error within ±0.20°. Therefore, this invention includes not only crystals with completely uniform diffraction angles in powder X-ray powder diffraction, but also crystals with uniform diffraction angles within an error range of ±0.20°. The crystalline form of the compounds of formula (I) of this invention is not limited to crystals having the same X-ray powder diffraction pattern as shown in the accompanying drawings; any crystal having a substantially identical X-ray powder diffraction pattern as shown in the accompanying drawings is within the scope of this invention.

[0164] It should be understood that different types of equipment or different testing conditions may yield slightly different DSC spectra and endothermic transition temperature readings. DSC data can reflect changes in the state of matter; endothermic peaks can indicate that the substance has undergone dehydration or desolvation, or crystal transformation, or melting, etc.; when reflecting the molten state, the corresponding temperature is generally understood as the melting point of the substance. This value will be affected by compound purity, sample weight, heating rate, particle size, and calibration and maintenance of the testing equipment. Those skilled in the art will understand that the temperature at which a substance changes from a solid to a liquid state is usually a temperature range, not a fixed point value; therefore, the temperature corresponding to the endothermic peak or the melting point of the substance can be characterized by either the onset value, the peak value, or other reasonable values. The maximum endothermic transition temperature of the crystal form can be within the range of ±5.0℃, preferably within ±2.0℃, of the specific values ​​disclosed above.

[0165] This invention also employs thermogravimetric analysis (TGA) to analyze the relationship between the degree of decomposition, sublimation, or evaporation of the crystal form (weight loss) and temperature. It should be understood that the values ​​obtained for the same crystal form may have some error due to factors such as sample purity, particle size, different types of equipment, and different testing methods. The temperature at which the crystal form decomposes, sublimates, or evaporates can be within ±5.0℃ of the specific values ​​disclosed above, for example, within ±2.0℃.

[0166] The "stability" of a crystal form includes "chemical stability" and / or "physical stability." "Chemical stability" refers to the degree to which the crystal form undergoes degradation reactions under certain temperature, humidity, and light conditions, reflecting its stability under storage conditions. "Physical stability" refers to the degree to which the crystal form undergoes a solid-state transformation under specific conditions, such as transformation into another crystal form under conditions of high temperature, high humidity, grinding, tableting, solvent removal, or solvent adsorption. Therefore, "stability" can, to some extent, reflect the stability of the crystal form during its use in formulations and other processes.

[0167] Regarding the description of hygroscopic characteristics and the definition of hygroscopic weight gain (Chinese Pharmacopoeia 2020 Edition, General Chapter 9103, Guidelines for Hygroscopicity Testing of Drugs):

[0168] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0169] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15.0%;

[0170] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15.0% but not less than 2.0%;

[0171] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2.0% but not less than 0.2%;

[0172] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0173] Hygroscopicity directly affects the physicochemical stability of drugs; high hygroscopicity easily leads to chemical degradation and crystal form transformation. Furthermore, high hygroscopicity reduces drug flowability, thus affecting processing techniques. Moreover, drugs with high hygroscopicity require low humidity during production and storage, placing higher demands on production and resulting in higher costs. More importantly, high hygroscopicity can easily cause changes in the content of active ingredients in drugs, affecting drug quality.

[0174] Changes in crystal form can alter drug absorption and affect bioavailability. Changes in bioavailability can then lead to changes in drug efficacy or trigger toxic side effects. Good stability ensures that drugs are stored with minimal impurities. Crystal forms possess good physicochemical stability, guaranteeing consistent and controllable quality between the active pharmaceutical ingredient and the finished product, minimizing changes in drug quality caused by changes in crystal form or the formation of impurities.

[0175] In this invention, the "stirring" is performed using conventional methods in the art, such as magnetic stirring or mechanical stirring, with a stirring speed of 50-1800 rpm. Magnetic stirring is preferably performed at 300-900 rpm, and mechanical stirring is preferably performed at 100-300 rpm.

[0176] The "drying" can be carried out at room temperature or higher. The drying temperature is from room temperature to approximately 60°C, or to 50°C, or to 40°C. The drying time can be 2-48 hours, or overnight. Drying is carried out in a fume hood, forced-air oven, or vacuum oven. The crystalline structure of the present invention can be prepared by various methods, including crystallization or recrystallization from a suitable solvent, sublimation, growth from a melt, solid-state transformation from another phase, crystallization from a supercritical fluid, and jet spraying, etc. Techniques for crystallizing or recrystallizing the crystalline structure from a solvent mixture include solvent evaporation, lowering the temperature of the solvent mixture, crystallization of a supersaturated solvent mixture of the molecule and / or salt, lyophilization of the solvent mixture, and adding an antisolvent to the solvent mixture, etc.

[0177] The term "antisolvent" refers to a solvent that is miscible with the solvent but cannot dissolve the solute.

[0178] "wt%" represents the percentage by mass.

[0179] In this invention, the compound of formula (I), API, and raw material of compound (I) all refer to the same substance, namely 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-(7-(4-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)but-3-yn-1-yl)-7-azaspiro[3.5]nonane-2-yl)-1H-pyrazole-4-yl)pyrazole[1,5-a]pyrimidine-3-amide, which can be obtained by the preparation method described in Example 1.

[0180] In this invention, the solvates (including hydrates) generally refer to the situation where solvent molecules have entered the interior of the crystal lattice. Based on the interaction force between the solvent and the main compound after entering the crystal lattice, solvates are classified into independent site type, channel type, and complex type. Solvent molecules remaining on the crystal surface are not considered solvates.

[0181] [Corrected according to Rule 91, 20.01.2026] Microstructural characteristics of independent-site solvates (also known as hole-type solvates): Solvent molecules within the crystal lattice only interact with drug compound molecules; there are no direct interactions between solvent molecules. The type can be determined by combining the characteristics of various analytical spectra. The morphological characteristics of independent-site solvates in their DSC / TGA spectra are: a) "sharper" desolventization peaks in the DSC spectrum; b) "relatively rapid" mass loss in the TGA spectrum; c) the desolventization temperature is not significantly related to the boiling point of the solvent molecules. Simultaneously, the hydrogen bonding between solvent molecules and the host compound molecules plays a crucial role in maintaining the stability of the entire unit cell structure. Therefore, during the desolventization process, these solvates generally undergo amorphous formation or crystal transformation accompanied by the collapse of the original unit cell structure.

[0182] Microstructural characteristics of channel-type solvates: Solvent molecules exist in the tunnels of the unit cell. Solvent molecules interact with each other along a certain direction, forming a chain-like arrangement to create independent "channel" structures. This channel structure can dynamically "close" and "open" with changes in external temperature / humidity. When the solvent channel is open, excess solvent can be adsorbed or removed from the channel. Morphological characteristics of its DSC / TGA spectra: a: Broad desolventizing peaks in the DSC spectrum (bulging type); b: Slow desolventizing process in the TGA spectrum; c: Initial desolventizing temperature is lower than the solvent boiling point. In this type of solvate, solvent molecules fill the cavities of the crystal under the action of van der Waals forces and do not participate in the construction of the unit cell network. Therefore, the solvent content has little effect on the unit cell structure, meaning that the XRD pattern changes very little during the desolventizing process.

[0183] Microstructural characteristics of ionic solvates: Solvent molecules and metal cations form complexes through hydrogen bonding. Ionic solvates contain metal ions, and their DSC / TGA generally exhibit high desolventization temperatures, with a prominent endothermic dehydration peak on DSC and a narrow desolventization temperature range on TGA. The crystal structure changes after desolventization. A single solvate molecule may have two or more types.

[0184] Generally, anhydrous crystalline forms have advantages such as lower residual solvent levels or better stability compared to other solvates or amorphous forms, and therefore have higher medicinal value. In this invention, the free base crystalline form of the compound of formula (I) was screened using various preparation methods, including slurrying, 50°C suspension stirring, antisolvent dropwise addition, evaporation, cooling crystallization, and heating crystallization. The solvents used included water, alcohols, ketones, esters, ethers, alkyl groups, nitriles, sulfones, and haloalkyl groups. Extensive crystalline form screening was conducted, and the above-mentioned crystalline forms described in this invention were obtained, among which crystalline form A is an anhydrous crystalline form with good physicochemical properties.

[0185] By selecting appropriate salt forms through salt form screening, important properties of APIs such as solubility, dissolution, bioavailability, hygroscopicity, taste, and physicochemical stability can be adjusted. In this invention, compounds of formula (I) are screened for salt forms with various acids, such as hydrochloric acid, maleic acid, p-toluenesulfonic acid, methanesulfonic acid, hydrobromic acid, succinic acid, L-malic acid, L-tartaric acid, benzenesulfonic acid, phosphoric acid, gentian acid, oxalic acid, fumaric acid, and citric acid. The solvents used include water, alcohols, ketones, esters, ethers, alkyl groups, and nitrile groups. Extensive screening of salts and their crystal forms was conducted. Among them, the p-toluenesulfonate and methanesulfonate of compounds of formula (I) described in this invention yield crystal forms with good physicochemical properties. Attached Figure Description

[0186] Figure 1. XRPD diffraction pattern of crystal form A.

[0187] Figure 2. TGA-DSC spectrum of crystal form A.

[0188] Figure 3. DVS pattern of crystal form A.

[0189] Figure 4. XRPD diffraction pattern of crystal form B.

[0190] Figure 5. TGA-DSC spectrum of crystal form B.

[0191] Figure 6. XRPD diffraction pattern of crystal form C.

[0192] Figure 7. TGA-DSC spectrum of crystal form C.

[0193] Figure 8. XRPD diffraction pattern of crystal form D.

[0194] Figure 9. TGA-DSC spectrum of crystal form D.

[0195] Figure 10. XRPD diffraction pattern of crystal form E.

[0196] Figure 11. TGA-DSC spectrum of crystal form E.

[0197] Figure 12 XRPD diffraction pattern of crystal form F.

[0198] Figure 13. TGA-DSC spectrum of crystal form F.

[0199] Figure 14. XRPD diffraction pattern of crystal form G.

[0200] Figure 15. TGA-DSC spectrum of crystal form G.

[0201] Figure 16. XRPD diffraction pattern of p-toluenesulfonate crystal form I.

[0202] Figure 17. TGA-DSC spectrum of p-toluenesulfonate crystal form I.

[0203] Figure 18. DVS spectrum of p-toluenesulfonate crystal form I.

[0204] Figure 19 Comparison of p-toluenesulfonate crystal forms I with different water contents.

[0205] Figure 20. Ellipsoidal diagram of single crystal structure of p-toluenesulfonate I.

[0206] Figure 21 shows a comparison of XRPD before and after heating of toluenesulfonate crystal form I.

[0207] Figure 22 XRPD diffraction pattern of p-toluenesulfonate crystal form II.

[0208] Figure 23. TGA-DSC spectrum of p-toluenesulfonate crystal form II.

[0209] [Corrected according to Rule 91 20.01.2026] Figure 24 XRPD pattern of methanesulfonate crystal form III.

[0210] Figure 25. TGA-DSC spectrum of methanesulfonate crystal form III.

[0211] Figure 26 shows the concentration-dependent distribution of the inflammatory factor IL-5 in mouse plasma after three PO dosage formula (I) compounds.

[0212] Figure 27 shows the concentration-dependent distribution of the inflammatory factor MCP-1 in mouse plasma after three PO dosage formula (I) compounds.

[0213] [Corrected according to Rule 91 20.01.2026] Figure 28 shows the concentration dependence of inflammatory factor IL-5 in mouse peritoneal lavage fluid after three PO dosage formula (I) compounds.

[0214] [Corrected according to Rule 91 20.01.2026] Figure 29 shows the concentration dependence of the inflammatory factor MCP-1 in the peritoneal lavage fluid of mice after three PO dosage formula (I) compounds.

[0215] Figure 30 shows the in vivo degradation of IRAK4 in the spleen of mice after three PO dosage formula (I) compounds. Detailed Implementation

[0216] The present invention will be described in detail below through embodiments, but this does not imply any adverse limitations on the invention. All technical solutions implemented based on the above description of the present invention fall within the scope of the present invention. The present invention provides a general and / or specific description of the materials and test methods used in the experiments; unless otherwise specified, the term "room temperature" in the present invention has a commonly known meaning in the art, specifically referring to 15–35°C, preferably 20–30°C, and more preferably 20–25°C.

[0217] The instruments and analytical methods used in this invention

[0218] (1) X-ray powder diffraction (XRPD)

[0219] Solid samples were analyzed using a Malvern-Panaco Aeris X-ray benchtop diffractometer. An appropriate amount of fine powder was taken, placed in the groove of the sample holder, and pressed into a flat and dense plane. The XRPD measurement parameters are shown in the table below.

[0220] XRPD test parameters

[0221] (2) TGA-DSC Simultaneous Thermal Analyzer

[0222] Thermogravimetric-differential scanning calorimetry (TGC) analysis of solids was performed using a Mettler Toledo simultaneous thermal analyzer. A suitable amount of the sample was placed in a crucible using a small spoon, ensuring even distribution, and its weight was measured. The sample was heated according to the parameters listed in the table, and the data were analyzed using STARE.

[0223] TGA-DSC Analysis Method Parameters

[0224] (3) Dynamic moisture adsorption-desorption analysis (DVS)

[0225] The hygroscopicity of the samples was determined using a DVS Intrinsic dynamic moisture adsorption analyzer. The samples were placed in a pre-peeled sample basket, and the instrument automatically weighed them. The samples were then analyzed according to the parameters in the table below.

[0226] DVS Analysis Method Parameters

[0227] (4) Nuclear magnetic resonance hydrogen spectrum ( 1 H-NMR)

[0228] The NMR measurements were performed using a Bruker AVANCE NEO 400 NMR instrument, with deuterated dimethyl sulfoxide (DMSO-d6) as the solvent.

[0229] (5) High Performance Liquid Chromatography (HPLC)

[0230] The HPLC determination was performed using a Waters e2695 high-performance liquid chromatograph.

[0231] (6) Karl Fischer moisture titration (KF) test

[0232] (a) Instruments and equipment

[0233] Analytical balance (Sartorius, MSE125P), Karl Fischer moisture analyzer (Beijing Pioneer Technology Co., Ltd., ZDJ400).

[0234] (b) Reagents and solutions

[0235] Water, Fischer solution, methanol-formamide (1:1) solution

[0236] (c) Operation process

[0237] The test was conducted according to the moisture determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0832, Method 1).

[0238] Calibration: Take an appropriate amount of water, weigh it accurately, and perform three calibrations to determine the titer of the titrant used; then perform a re-standardization to confirm the recovery rate, which should be within the range of 97.5% to 102.5%.

[0239] Test: Weigh 0.1 to 0.2 g of this product and add it to a titration cup containing a methanol-formamide (1:1) solution. Measure the solution after the sample has completely dissolved.

[0240] (d) Calculation formula:

[0241] In the formula: A: Volume of Fischer solution consumed by the test sample (ml)

[0242] F: 1 mL of Fischer solution is equivalent to the weight (mg) of water.

[0243] W: Weight of the test sample (g).

[0244] (7) Microcrystalline Electron Diffraction (MicroED)

[0245] MicroED Instrument Method

[0246] Procedure: A small amount of microcrystalline sample was taken and uniformly dispersed onto a cryo-electron microscope (CEM) grid. The prepared grid was placed at the front end of a liquid nitrogen cryo-sample holder (Model 915, Gatan), which was then positioned on a dedicated cryo-transfer stage. Liquid nitrogen was added to the groove of the cryo-transfer stage and the Dewar flask at the end of the cryo-sample holder, ensuring the crystals on the grid were completely immersed in the liquid nitrogen. The temperature was allowed to gradually stabilize to the liquid nitrogen temperature. Finally, the sample holder was quickly inserted into the Talos F200C CEM. During the test, the microcrystalline sample was irradiated with a parallel electron beam (NanoProbe mode). A goniometer continuously and uniformly rotated the microcrystalline sample at a speed of 2.0° / second, while a Ceta-D camera collected MicroED data from the microcrystalline sample in continuous exposure mode. The MicroED data collection angle was 80-100°, and the exposure time for a single diffraction image was 0.5 seconds. Thirteen sets of relatively good MicroED data were selected. XDS software was used to index diffraction points, determine unit cell parameters, and integrate diffraction intensity. The XSCALE program was then used to merge the data, resulting in the final diffraction data file. The merged diffraction data file contains a total of 131,590 diffraction points and 12,853 independent diffraction points. During crystal structure analysis, the atomic scattering factor of electrons was used to calculate the theoretical structure factor, and SHELXT (Intrinsic phasing algorithm) software was used to perform structure analysis, obtaining the initial structural model. SHELXL (least squares method) was used to refine the structure, and the positions of all hydrogen atoms were obtained by combining Fourier difference plots with calculations.

[0247] [Corrected according to Rule 91, 20.01.2026] Example 1, Preparation of compound (I):

[0248] Synthesis of intermediate A

[0249] Step 1: Under argon protection and at room temperature, Aa (8.32 g, 36.88 mmol) and (1R,4R)-2-oxa-5-azabicyclo[2,2,1]heptane hydrochloride (5 g, 36.88 mmol) were dissolved in acetonitrile (60 mL), and N,N-diisopropylethylamine (14.3 g, 110.63 mmol) was slowly added dropwise with stirring. The reaction mixture was then stirred overnight at 60 °C. LC-MS showed complete consumption of the starting material and formation of the sole product, Ab. Water (100 mL) was added to the cooled reaction mixture, followed by extraction with ethyl acetate (3 × 50 mL). The combined organic phases were washed with saturated brine (3 × 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the residue was separated by silica gel column chromatography to obtain Ab (10.1 g). 1H NMR (400MHz, CDCl3) δ8.32-8.20(m,2H),6.12(brs,1H),5.45(brs,1H),4.82-4.71(m,1H),4. 42-4.26(m,2H),4.02-3.84(m,2H),3.51-3.37(m,2H),2.12-1.87(m,2H),1.41-1.39(m,3H).

[0250] Step 2: At room temperature, Ab (10.1 g, 35.03 mmol) was dissolved in anhydrous methanol / water (100 mL / 25 mL), and lithium hydroxide monohydrate (8.82 g, 210.19 mmol) was added with stirring. The reaction mixture was then stirred overnight at 60 °C. LC-MS showed a trace amount of the starting material remaining, and Ac was the main product. The cooled reaction mixture was diluted with water (100 mL), and the pH was adjusted to 5 with dilute hydrochloric acid (1 M) in an ice-water bath. The resulting mixture was extracted with ethyl acetate (3 × 50 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give Ac (8.7 g). 1 HNMR(400MHz, CDCl3)δ8.36-8.28(m,2H),6.40-6.17(m,1H),5.22,4.82(s, 1H),4.68(s,1H),3.99-3.90(m,2H),3.72-3.51(m,2H),2.13-2.03(m,2H).

[0251] Step 3: 1H-pyrazole-5-carboxaldehyde (58.7 g, 610.89 mmol) and TsOH·H2O were dissolved in DCM (580 mL), cooled to 0 °C, and DHP (61.6 g, 733.07 mmol) was slowly added. The ice bath was removed, and the mixture was allowed to react overnight at room temperature. TEA (23.5 mL) was added to the reaction solution, and the mixture was concentrated by rotary evaporation. The residue was purified by silica gel column chromatography to obtain Ad (88 g). 1 H NMR(400MHz, CDCl3) δ10.00(s,1H),7.68(dd,J=2.6,0.6Hz,1H),6.83(d,J=2.8Hz,1H),5.4 9-5.45(m,1H),4.09-4.05m,1H),3.77-3.70(m,1H),2.16-2.03(m,3H),1.77-1.63(m,3H).

[0252] Step 4: Dissolve Ad (88g, 488.33mmol) in DCM (500mL), add DAST (157g, 976.65mmol) dropwise at 0℃, remove ice bath and let the reaction proceed overnight at a low temperature. TLC (PE / EtOAc = 10 / 1) showed that the reaction was not complete. The reaction solution was slowly quenched with NaHCO3 (3L), the organic phase was separated, and the aqueous phase was further extracted with DCM (3×1L). The organic phases were combined, dried over Na2SO4, filtered, concentrated by rotary evaporation, and the residue was purified by silica gel column chromatography to obtain Ae (69.65g). 1 H NMR (400MHz, CDCl3) δ7.63 (d, J = 2.4Hz, 1H), 6.70 (t, J = 55.0Hz, 1H), 6.51 (d, J = 2.4Hz, 1H), 5. 39-5.35(m,1H),4.07-4.03(m,1H),3.75-3.65(m,1H),2.15-1.97(m,3H),1.74-1.57(m,3H).

[0253] Step 5: Dissolve Ae (25.7 g, 127.10 mmol) in MeOH (254 mL), add HCl (4 M in 1,4-dioxane) (254 mL, 1.016 mol) at 0 °C, remove the ice bath and let the reaction proceed overnight at room temperature. Monitor the reaction by TLC until complete. After concentrating the reaction solution, add saturated NaHSO3 (420 mL) and EtOAc (420 mL), stir at room temperature for 1 h, extract with EtOAc (3 × 200 mL), combine the organic phases, dry with Na2SO4, and concentrate by rotary evaporation to obtain Af (16.8 g). 1 H NMR (400MHz, CDCl3) δ7.65-7.63 (m, 1H), 6.80 (t, J = 55.6Hz, 1H), 6.56-6.54 (m, 1H).

[0254] Step 6: Dissolve Ag (14.0 g, 118.56 mmol) in concentrated H₂SO₄ (140 mL), heat to 0°C on ice, add HNO₃ (65%, 40.23 g, 414.95 mmol) dropwise, slowly raise the temperature to room temperature, then gradually raise it to 60°C, and react overnight at 115°C. Cool the reaction solution to room temperature, pour it into ice water (1 L), and extract with EtOAc (3 × 1 L). Combine the organic phases, dry them in Na₂SO₄, concentrate by rotary evaporation, and purify to obtain Ag (7.79 g). 1 H NMR (400MHz, CDCl3) δ8.46 (s, 1H), 7.35 (t, J = 53.6Hz, 1H).

[0255] Step 7: 2-O-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (9.57 g, 39.99 mmol) was dissolved in anhydrous methanol (100 mL) at room temperature. Sodium borohydride (1.82 g, 47.99 mmol) was slowly added under stirring in an ice-water bath. The reaction mixture was slowly brought to room temperature and stirred for 1 hour. Thin-layer chromatography showed that the starting material reacted completely. A saturated ammonium chloride aqueous solution (100 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (3 × 100 mL). The combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the residue was separated by silica gel column chromatography to obtain Ah (9.14 g). 1 H NMR (400MHz, CDCl3) δ4.37-4.28(m,1H),3.35-3.27(m,4H),2.31-2.25(m, 2H),1.80-1.78(m,1H),1.71-1.65(m,2H),1.54-1.47(m,4H),1.45(s,9H).

[0256] Step 8: At room temperature, Ah (9.14 g, 37.87 mmol) was dissolved in toluene (90 mL). Triphenylphosphine (14.90 g, 56.81 mmol), imidazole (5.16 g, 75.75 mmol), and iodine (14.42 g, 56.81 mmol) were added sequentially with stirring in an ice-water bath. The reaction mixture was slowly brought to room temperature and stirred for 1 hour, then heated to 60 °C and stirred for 1 hour. LC-MS showed that the starting material reacted completely. Water (200 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (3 × 200 mL). The combined organic phases were washed with saturated brine (400 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the residue was separated by silica gel column chromatography to obtain Ai (7.98 g). 1 H NMR (400MHz, CDCl3) δ4.54-4.45(m,1H),3.34-3.30(m,2H),3.29-3.25(m,2H),2.69 -2.63(m,2H),2.45-2.39(m,2H),1.69-1.65(m,2H),1.57-1.53(m,2H),1.44(s,9H).

[0257] Step 9: At room temperature, Ai (5 g, 14.24 mmol) and Ag (2.11 g, 12.94 mmol) were dissolved in N,N-dimethylformamide (50 mL), and potassium carbonate (3.58 g, 25.88 mmol) was added with stirring. The reaction mixture was then stirred overnight at 100 °C. LC-MS showed that the starting material reacted completely. The reaction mixture was cooled to room temperature, and water (100 mL) was added, followed by extraction with ethyl acetate (3 × 100 mL). The combined organic phases were washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the residue was separated by silica gel column chromatography to give Aj (3.8 g, yield: 76%). 1 H NMR (400MHz, DMSO-d6) δ9.13 (s, 1H), 7.32 (t, J = 53.0Hz, 1H), 5.05-5.00 (m, 1H), 3.32-3.29 (m, 2H),3.25-3.21(m,2H),2.44-2.38(m,2H),2.30-2.23(m,2H),1.60-1.53(m,4H),1.39(s,9H).

[0258] Step 10: Dissolve Aj (3.8 g, 9.83 mmol) in tetrahydrofuran (60 mL) at room temperature, then add 10% wetted palladium on carbon (1.9 g) at room temperature. The reaction mixture is then stirred at 30 °C under hydrogen balloon protection for 3 hours. LC-MS showed that the starting material reacted completely. Filter the reaction mixture, and wash the filter cake with ethyl acetate (100 mL). Concentrate the combined filtrates to give the desired product Ak (3.4 g). MS (ESI) M / Z: 357 [M+H] + .

[0259] Step 11: At room temperature and under argon protection, Ak (2.00 g, 5.61 mmol) and Ac (1.50 g, 5.76 mmol) were dissolved in acetonitrile (40 mL). 1-Methylimidazole (1.60 mL, 20.07 mmol) and N,N,N',N'-tetramethylchloroformamidin hexafluorophosphate (TCFH, 2.40 g, 8.55 mmol) were added with stirring in an ice-water bath. The argon atmosphere was purged three times. The reaction mixture was then stirred overnight at 50 °C under argon protection. LC-MS showed complete reaction of the starting materials. A saturated sodium bicarbonate aqueous solution (40 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (3 × 40 mL). The combined organic phases were washed with saturated brine (2 × 40 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography to obtain the desired product Al (3.40 g). MS(ESI)M / Z:599.1[M+H] + .

[0260] Step 12: Al (3.40 g, 5.68 mmol) was dissolved in dichloromethane (40 mL) at room temperature. Hydrochloric acid / 1,4-dioxane solution (4 M, 14 mL, 56 mmol) was added with stirring in an ice-water bath. The reaction mixture was then stirred at room temperature for 2 hours. LC-MS showed that the starting material reacted completely. The reaction mixture was concentrated to obtain the desired crude product A (3.87 g, hydrochloride). MS (ESI) M / Z: 499.6 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.51(d,J=5.2Hz,1H),8.89(s,2H),8.79(d,J=8.0Hz,1H),8.42(d,J=4 .0Hz,1H),8.26(d,J=5.6Hz,1H),7.14(t,J=53.6Hz,1H),6.89-6.45(m,1H),5.28-5.08(m,1H), 5.00-4.92(m,1H),4.80-4.74(m,1H),3.87-3.73(m,2H),3.65-3.43(m,2H),3.04-2.95(m,4H), 2.47-2.41(m,2H),2.30-2.24(m,2H),2.06-1.92(m,2H),1.88-1.84(m,2H),1.82-1.79(m,2H).

[0261] Synthesis of intermediate B:

[0262] Step 1: At room temperature, glutarimide (100 g, 884.0 mmol) was placed in a 500 mL sealed container, chloroform (200 mL) was added, and liquid bromine (141 g, 884.0 mmol) was slowly added while stirring. The container was sealed and the reaction mixture was stirred overnight at 105 °C. LC-MS showed that the starting material had basically reacted completely, and the main product was Bb. The reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in ethyl acetate (1 L), washed with saturated sodium bicarbonate aqueous solution (500 mL), washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the residue was separated by silica gel column chromatography to obtain the desired product Bb (78 g). MS (ESI) M / Z: 192.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ11.05 (s, 1H), 4.88 (t, J = 4.8Hz, 1H), 2.56-2.52 (m, 2H), 2.49-2.43 (m, 1H), 2.19-2.10 (m, 1H).

[0263] Step 2: At room temperature, p-methoxybenzyl alcohol (20.6 g, 148.9 mmol) was dissolved in dry tetrahydrofuran (500 mL). Triphenylphosphine (39.1 g, 148.9 mmol) and Bb (26 g, 135.4 mmol) were added with stirring. The reaction solution was then cooled to 0 °C, and diethyl azodicarbonate (25.9 g, 148.9 mmol) was slowly added dropwise under argon protection. After the addition was complete, the reaction solution was allowed to warm naturally to room temperature and stirred overnight. LC-MS showed that the starting material reacted completely. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography to obtain the desired product Bc (29.7 g). 1 H NMR (400MHz, CDCl3) δ7.34-7.27(m,2H),6.87-6.78(m,2H),4.90(dd,J=37.9,13.8Hz,2H),4.74-4. 68(m,1H),3.78(s,3H),3.10-2.96(m,1H),2.80-2.69(m,1H),2.42-2.30(m,1H),2.27-2.17(m,1H).

[0264] Step 3: At room temperature, Bd (19.8 g, 87.4 mmol) was dispersed in dry tetrahydrofuran (400 mL), stirred, and cooled to 0 °C. Potassium tert-butoxide (16.7 g, 148.5 mmol) was added under argon protection. The reaction mixture was stirred at this temperature for 1 hour, then heated to 45 °C and a dry tetrahydrofuran solution (300 mL) of Bc (30.0 g, 96.1 mmol) was slowly added dropwise over 6 hours. After the addition was complete, the reaction mixture was stirred at 45 °C for 3 hours. LC-MS showed that the starting material had reacted almost completely. The reaction mixture was cooled to 0 °C, quenched slowly with saturated brine (300 mL), and separated. The organic phase was washed once more with saturated brine (300 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography to obtain the desired product Be (18.3 g). 1 H NMR (400MHz, DMSO-d6) δ7.29-7.15(m,3H),7.13-7.04(m,1H),6.94(t,J=8.0Hz,1H),6.89-6.81(m,2H),5.57(dd,J=13.0 ,5.4Hz,1H),4.79(q,J=14.3Hz,2H),3.72(s,3H),3.64(s,3H),3.13-2.94(m,1H),2.88-2.65(m,2H),2.14-1.99(m,1H).

[0265] Step 4: Dissolve Be (18.3 g, 19.9 mmol) in toluene (140 mL) and methanesulfonic acid (70 mL) at room temperature, heat to 120 °C and stir for 5 hours. LC-MS showed that the starting material reacted completely. Cool the reaction solution to room temperature and concentrate it. Slowly add the residue dropwise to ice water (1 L), filter, and dry to obtain the desired product B (10.5 g). 1 H NMR (400MHz, DMSO-d6) δ11.13(s,1H),7.24(d,J=8.0Hz,1H),7.17(d,J=8.0Hz,1H),6.98(t,J=8.2Hz,1 H),5.41(dd,J=12.4,5.2Hz,1H),3.64(s,3H),2.96-2.82(m,1H),2.77-2.59(m,2H),2.10-1.98(m,1H).

[0266] Preparation of compound (I)

[0267] Step 1: 3-Butyn-1-ol (0.5 g, 7.13 mmol) was dissolved in dichloromethane (10 mL) at room temperature. p-Toluenesulfonyl chloride (1.63 g, 8.56 mmol) and pyridine (850 mg, 10.70 mmol) were added sequentially with stirring in an ice-water bath. The reaction mixture was then stirred at room temperature for 16 hours. Thin-layer chromatography showed complete reaction of the starting material. A saturated ammonium chloride aqueous solution (20 mL) was added to the reaction mixture, followed by extraction with ethyl acetate (2 × 50 mL). The combined organic phases were washed with saturated brine (3 × 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography to obtain the desired product 22a (860 mg). 1 H NMR (400MHz, CDCl3) δ7.80(d,J=7.6Hz,2H),7.36(d,J=8.0Hz,2H),4.10(t,J=7.0Hz,2H),2.57-2.52(m,2H),2.45(s,3H),1.99(t,J=2.8Hz,1H).

[0268] Step 2: 22a (201.8 mg, 0.90 mmol) was dissolved in N,N-dimethylformamide (4 mL) at room temperature. A (300 mg, 0.60 mmol) and cesium carbonate (586.5 mg, 1.80 mmol) were added with stirring. The reaction mixture was then stirred overnight at 60 °C. LC-MS showed that the starting material reacted completely. Water (30 mL) was added to the cooled reaction mixture, followed by extraction with ethyl acetate (3 × 30 mL). The combined organic phases were washed with saturated sodium bicarbonate aqueous solution (2 × 30 mL), then with saturated brine (2 × 30 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography to obtain the desired product 22b (123 mg). MS (ESI) M / Z: 551.4 [M+H] + ;

[0269] 1 H NMR (400MHz, CDCl3) δ9.61 (s, 1H), 8.43 (t, J = 8.4Hz, 2H), 8.32 (d, J = 7.6Hz, 1H), 6.78 (t,J=54.0Hz,1H),6.13(d,J=7.6Hz,1H),5.46(s,1H),4.81(d,J=10.4Hz,1H),4.75-4 .67(m,1H),4.02-3.94(m,2H),3.62-3.44(m,2H),2.61(t,J=7.6Hz,2H),2.55-2.35(m ,8H),2.33-2.24(m,2H),2.14-2.09(m,1H),1.99(t,J=2.6Hz,2H),1.78-1.70(m,5H).

[0270] Step 3: 22b (123 mg, 0.22 mmol) and B (50 mg, 0.15 mmol) were dissolved in dry N,N-dimethylformamide (2 mL) at room temperature under argon protection. Cesium carbonate (122.2 mg, 0.38 mmol), palladium dichloride bis(triphenylphosphine) (16 mg, 0.022 mmol), and cuprous iodide (9.0 mg, 0.045 mmol) were added sequentially under argon protection. The reaction was then stirred at 80 °C for 5 hours under argon protection. LC-MS showed the formation of product (I). The cooled reaction solution was filtered, and the filter cake was washed with ethyl acetate (20 mL). The filtrate was extracted with ethyl acetate (2 × 20 mL) after adding saturated sodium bicarbonate aqueous solution (10 mL). The combined organic phases were washed with saturated sodium chloride aqueous solution (5 × 20 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by thin-layer preparative chromatography to obtain the desired product (I) (6.9 mg).

[0271] The obtained product was purified by silica gel column chromatography, followed by gradient elution with dichloromethane / methanol, and then rotary evaporation under reduced pressure to obtain a white powder, which was found to be amorphous by XRD.

[0272] MS(ESI)M / Z:808.6[M+H] + ; 1 H NMR (400MHz, CDCl3) δ9.61 (s, 1H), 8.43 (t, J = 10.8

[0273] Hz,2H),8.32(d,J=8.0Hz,1H),8.10(s,1H),7.12(d,J=8.0Hz,1H),7.00-6.94(m,1H) ,6.78(t,J=54.6Hz,1H),6.71(d,J=8.0Hz,1H),6.13(t,J=7.6Hz.1H),5.46(s,1H),5. 25-5.16(m,1H),4.83-4.68(m,2H),4.01-3.95(m,2H),3.80(s,3H),3.62-3.48(m,2H) ,2.99-2.96(m,1H),2.88-2.70(m,2H),2.68(s,4H),2.50-2.35(m,11H),1.75(s,4H).

[0274] Example 2 Preparation of the free alkali crystal form of compound (I)

[0275] 2.1 Preparation of Crystal Form A

[0276] Weigh 0.10 g of the amorphous free base of compound (I) and add it to 5 mL of methanol / water (v / v, 5:1) preheated to 50 °C. Stir until dissolved quickly, and a solid begins to precipitate after about 5 minutes of stirring. Continue stirring for 30 minutes, then cool to room temperature and stir for 60 minutes. Filter to obtain a white solid, and dry under vacuum at 40 °C for 4 hours to obtain crystal form A. Its XPRD spectrum is shown in Figure 1.

[0277] The TGA-DSC spectrum is shown in Figure 2. The spectrum shows that crystal form A has an endothermic peak at 204.8℃±2℃; and loses 0.7% of its weight at 35-180℃.

[0278] The DVS diagram is shown in Figure 3.

[0279] Crystal form A 1H-NMR (400MHz, DMSO-d6): δ (ppm) 11.13 (s, 1H), 9.51-9.50 (d, J = 5.2Hz, 1H), 8.80-8.78 (dd, J = 8.0, 1.6Hz, 1H), 8.41-8.40 (d, J = 4.0H) z,1H),8.27-8.26(d,J=5.6Hz,1H),7.29-6.97(m,4H),6.88-6.86(d,J=8.0Hz,0.5H),6.47-6.45(d,J=8.0Hz,0.5H),5.41-5.37(dd, J=12.4、5.2Hz,1H),5.28(s,0.5H),5.08(s,0.5H),4.97-4.93(m,1H),4.80-4.76(d,J=16.0Hz,1H),3.83-3.73(m,2H),3.68(s,3H), 3.65-3.44(m,2H),2.95-2.81(m,1H),2.73-2.54(m,6H),2.47-2.26(m,6H),2.21-2.16(m,2H),2.06-1.96(m,3H),1.67-1.60(m,4H).

[0280] 2.2 Preparation of crystal form B

[0281] Weigh 21.3 mg of the amorphous free base of compound (I), add 3.0 mL of methanol, heat to 50 °C, stir and slurry for 1 day, centrifuge to obtain solid, place at 40 °C and vacuum dry for 4 h to obtain crystal form B, whose XPRD spectrum is basically shown in Figure 4.

[0282] The TGA-DSC spectrum is shown in Figure 5. The spectrum shows that crystal form B has an endothermic peak at 196.6℃±2℃ and loses 3.8% of its weight between 35℃ and 209℃.

[0283] Crystal form B 1H-NMR (400MHz, DMSO-d6): δ (ppm) 11.13 (s, 1H), 9.52-9.50 (d, J = 5.2Hz, 1H), 8.80-8.78 (dd, J = 8.0, 1.6Hz, 1H), 8.42-8.41 (d, J = 4.0Hz, 1 H),8.27-8.26(d,J=5.6Hz,1H),7.29-6.98(m,4H),6.88-6.86(d,J=8.0Hz,0.5H),6.47-6.45(d,J=8.0Hz,0.5H),5.42-5.37(dd,J=12.4 ,5.2Hz,1H),5.28(s,0.5H),5.08(s,0.5H),4.98-4.94(m,1H),4.80-4.75(d,J=16.0Hz,1H),3.84-3.73(m,2H),3.68(s,3H),3.65-3.44 (m,2H),3.17(s,3H),2.93-2.85(m,1H),2.76-2.61(m,5H),2.43-2.29(m,4H),2.25-2.14(m,2H),2.06-1.93(m,3H),1.81-1.53(m,4H).

[0284] 2.3 Preparation of Crystal Form C

[0285] Weigh 12.5 mg of the amorphous free base of compound (I) and add it to 0.6 mL of ethanol / water (v / v, 5:1) preheated to 50 °C. Stir to dissolve and continue stirring for 1 h. After stirring at room temperature for 1 h, the solid precipitates. After centrifugation, collect the solid and dry it under vacuum at 40 °C for 4 h to obtain crystal form C. Its XPRD spectrum is shown in Figure 6.

[0286] The TGA-DSC spectrum is shown in Figure 7. The spectrum shows that crystal form C has an endothermic peak at 195.7℃±2℃ and loses about 5.7% of its weight between 36 and 213℃.

[0287] 2.3 Preparation of crystal form D

[0288] Weigh 12.0 mg of the amorphous free base of compound (I) and add it to 0.6 mL of isopropanol / water (v / v, 5:1) preheated to 50 °C. Stir to dissolve and continue stirring for 1 h. After stirring at room temperature for 1 h, the solid precipitates. After centrifugation, collect the solid and dry it under vacuum at 40 °C for 4 h to obtain crystal form D. Its XPRD spectrum is basically shown in Figure 8.

[0289] The TGA-DSC spectrum is shown in Figure 9. The spectrum shows that crystal form D has an endothermic peak at 202.35℃±2℃ and loses about 7.1% of its weight between 36 and 217℃.

[0290] 2.4 Preparation of Crystal Form E

[0291] Weigh 12.3 mg of the amorphous free base of compound (I) and add it to 0.6 mL of acetone / water (v / v, 5:1) preheated to 50 °C. Stir to dissolve and continue stirring for 1 h. After stirring at room temperature for 1 h, the solid precipitates. After centrifugation, collect the solid and dry it under vacuum at 40 °C for 4 h to obtain crystal form E. Its XPRD spectrum is basically shown in Figure 10.

[0292] The TGA-DSC spectrum is shown in Figure 11. The spectrum shows that crystal form E has an endothermic peak at 197.72℃±2℃ and loses about 5.2% of its weight between 95 and 214℃.

[0293] 2.5 Preparation of crystal form F

[0294] Weigh 12.3 mg of the amorphous free base of compound (I) and add it to 0.6 mL of acetonitrile / water (v / v, 5:1) preheated to 50 °C. Stir to dissolve and continue stirring for 1 h. After stirring at room temperature for 1 h, a solid precipitates. Centrifuge to collect the solid and dry it under vacuum at 40 °C for 4 h to obtain crystal form F. Its XPRD spectrum is shown in Figure 12.

[0295] The TGA-DSC spectrum is shown in Figure 13. The spectrum shows that crystal form F has an endothermic peak at 203.24℃±2℃ and loses about 4.4% of its weight at 36~215℃.

[0296] Crystal form F 1H-NMR (400MHz, DMSO-d6): δ (ppm) 11.13 (s, 1H), 9.51-9.50 (d, J = 5.2Hz, 1H), 8.80-8.78 (dd, J = 8.0, 1.6Hz, 1H), 8.42-8.41 (d, J = 4.0Hz, 1 H),8.27-8.26(d,J=5.6Hz,1H),7.29-6.98(m,4H),6.88-6.86(d,J=8.0Hz,0.5H),6.47-6.45(d,J=8.0Hz,0.5H),5.42-5.37(dd,J=12.4 ,5.2Hz,1H),5.28(s,0.5H),5.08(s,0.5H),4.98-4.94(m,1H),4.80-4.76(d,J=16.0Hz,1H),3.84-3.73(m,2H),3.68(s,3H),3.65-3.44 (m,2H),2.91-2.85(m,1H),2.73-2.55(m,6H),2.47-2.26(m,6H),2.21-2.17(m,2H),2.08(s,2H),2.06-1.96(m,3H),1.68-1.63(m,4H).

[0297] 2.6 Preparation of crystal form G

[0298] Weigh 20.0 mg of the amorphous free base of compound (I), add 1.0 mL of acetonitrile, heat to 50 °C, stir and slurry for 5 days, centrifuge to collect the solid, place it at 40 °C and vacuum dry for 4 h to obtain white solid crystal form G, whose XPRD spectrum is basically shown in Figure 14.

[0299] The TGA-DSC spectrum is shown in Figure 15. The spectrum shows that crystal form G has an endothermic peak at 199.1℃±2℃, and loses about 2.7% of its weight between 35 and 118℃ and about 2.6% of its weight between 118 and 221℃.

[0300] Example 3 Preparation of the salt of compound (I) and its crystal form

[0301] 3.1 Preparation of p-toluenesulfonic acid crystal form I of compound (I)

[0302] Method 1:

[0303] Weigh 25.0 mg of the amorphous free base of compound (I), add 1.0 mL of acetonitrile and 5.6 mg of p-toluenesulfonic acid, stir and slurry at room temperature for 3 days, collect the solid by centrifugation, and dry it under vacuum at 40 °C for 4 h to obtain a white solid, which is p-toluenesulfonic acid crystal form I. Its XPRD spectrum is basically shown in Figure 16.

[0304] Method 2:

[0305] 10 mL of isopropanol, 4 mL of purified water, and 0.225 g of p-toluenesulfonic acid were added to the reaction flask. The mixture was stirred and heated to 60 °C. 1.00 g of the free base of compound (I) was added and stirred until almost completely dissolved. The solution was filtered through a 0.45 μm microporous filter to remove a small amount of insoluble matter. The supernatant was cooled to 50 °C, and 5 mg of p-toluenesulfonate crystal form I of compound (I) was added. The mixture was stirred at 50 °C for 0.5 h. The temperature was then lowered to 40 °C, and 30 mL of isopropanol was slowly added dropwise over 2 h. After the addition was completed, the temperature was slowly lowered to 10 °C over 3 h. The mixture was stirred at 10 °C for 1 h and then vacuum filtered. The solution was dried under vacuum at 40 °C for 4 h to obtain a white solid, p-toluenesulfonic acid crystal form I. Its XPRD spectrum is shown in Figure 16.

[0306] The TGA-DSC spectrum is shown in Figure 17. The TGA spectrum shows a weight loss of about 2.1% at 35-134℃. This weight loss process is relatively gradual and there is no obvious step-like weight loss. At the same time, its DSC is accompanied by an endothermic peak. Crystal form I has a melting endothermic peak at 237.7℃±2℃.

[0307] Preparation of single crystal samples of p-toluenesulfonate I of compound (I):

[0308] The single crystal sample of p-toluenesulfonate I of compound (I) was obtained by slow evaporation of p-toluenesulfonate I of compound (I) in a mixed solvent of isopropanol and water. XRD diffraction showed that it was p-toluenesulfonate I. The sample was then subjected to MicroED detection to obtain the single crystal structure.

[0309] The actual moisture content measured by the Karl Fischer (KF) test was 2.7%.

[0310] DVS diagram 18 shows that the adsorption and desorption of water by p-toluenesulfonate crystal form I is reversible. The moisture absorption weight gain of p-toluenesulfonate crystal form I is 1.94% from RH 0% to RH 10%, corresponding to the gain of 1 equivalent of water molecules.

[0311] p-Toluenesulfonate crystal form I exhibits good stability under different humidity conditions. As the ambient humidity continues to increase, the moisture content of the sample will rise, and the water content of p-toluenesulfonate crystal form I can reach 5±0.5%. However, samples with different moisture contents all show the same XRD diffraction pattern (see Figure 19), indicating that p-toluenesulfonate crystal form I has good stability and the crystal form is not affected by changes in external humidity and its own moisture content.

[0312] Furthermore, based on the single-crystal structure data (see Figure 20), the water molecules in the lattice of this p-toluenesulfonate crystal form I do not form strong hydrogen bonds, and the water molecules in the lattice exist in the channel structure.

[0313] p-Toluenesulfonate crystal form I exhibits good thermal stability. XRD data (see Figure 21) show that after heating the crystal form I sample to 150℃ and removing water molecules at this temperature, the XRD pattern of the crystal form does not change significantly.

[0314] Compound (I) p-Toluenesulfonate crystal form I 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 11.14 (s, 1H), 9.53-9.51 (d, J = 5.2Hz, 1H), 9.25 (brs, 1H), 8.81-8.79 (dd, J = 8.0, 1.6Hz, 1H), 8.45-8.44 ( d,J=4.0Hz,1H),8.27-8.26(d,J=5.6Hz,1H),7.49-7.47(d,J=8.0Hz,2H),7.29-7.02(m,6H),6.89-6.87(d,J=8.0Hz,0.5H),6.48-6.46(d,J =8.0Hz, 0.5H), 5.43-5.39(dd, J = 12.4, 5.2Hz, 1H), 5.28(s, 0.5H), 5.09(s, 0.5H), 5.01-4.97(m, 1H), 4.80-4.75(d, J = 16.0Hz, 1H), 3.84-3.73(m, 2H), 3.65(s, 3H), 3.63-3.41(m, 5H), 3.09-2.86(m, 5H), 2.74-2.55(m, 3H), 2.35-2.26(m, 6H), 2.10-1.95(m, 5H), 1.85-1.77(m, 2H). NMR data showed that the molar ratio of compound (I) to p-toluenesulfonate was 1:1.

[0315] 3.2 Preparation of p-toluenesulfonate crystal form II of compound (I)

[0316] Weigh 30.6 mg of p-toluenesulfonate crystal form I of formula (I), add 2.5 mL of methanol / water (v / v, 1:1), heat to 60 °C and stir to dissolve. After passing through a 0.45 μm microporous filter, slowly cool the clear liquid to room temperature under magnetic stirring to precipitate solid. Continue cooling to 10 °C and stirring for 1 h, then centrifuge to collect the solid and place it in a vacuum dryer at 40 °C for 4 h to obtain p-toluenesulfonate crystal form II. Its X-ray powder diffraction pattern is shown in Figure 22.

[0317] The TGA-DSC spectrum is shown in Figure 23. The TGA spectrum shows a weight loss of about 2.3% at 36-169℃, accompanied by an endothermic peak; the crystal form has an endothermic peak at 223.2℃±2℃.

[0318] Compound (I) p-Toluenesulfonate crystal form II1 H-NMR (400MHz, DMSO-d6): δ (ppm) 11.13 (s, 1H), 9.52-9.51 (d, J = 5.2Hz, 1H), 9.24 (brs, 1H), 8.81-8.79 (dd, J = 8.0, 1.6Hz, 1H), 8.45-8.44 ( d,J=4.0Hz,1H),8.27-8.26(d,J=5.6Hz,1H),7.49-7.47(d,J=8.0Hz,2H),7.29-7.02(m,6H),6.89-6.87(d,J=8.0Hz,0.5H),6.48-6.46(d,J =8.0Hz,0.5H),5.43-5.38(dd,J=12.4,5.2Hz,1H),5.28(s,0.5H),5.09(s,0.5H),5.01-4.97(m,1H),4.80-4.75(d,J=16.0Hz,1H),3.84-3 .73(m,2H),3.65(s,3H),3.63-3.41(m,5H),3.09-2.86(m,5H),2.76- 2.55(m,3H),2.35-2.26(m,6H),2.10-1.95(m,5H),1.85-1.77(m,2H).

[0319] 3.3 Preparation of Methanesulfonate Crystal Forms I and II of Compound (I)

[0320] Method 1:

[0321] Weigh 25.0 mg of the amorphous free base of compound (I) into a small bottle, add 1.0 mL of acetonitrile, 0.1 mL of purified water and 3.7 mg of methanesulfonic acid, stir and slurry at room temperature for 1 day, centrifuge to collect the solid, and dry it under vacuum at 40 °C for 4 h to obtain a white solid, which is methanesulfonate crystal form IIⅠ. Its X-ray powder diffraction pattern is basically shown in Figure 24.

[0322] Method 2:

[0323] 10 mL of isopropanol, 4 mL of purified water, and 0.128 g of methanesulfonic acid were added to the reaction flask. The mixture was stirred and heated to 60 °C. 1.00 g of the free base of compound (I) was added and stirred until almost dissolved. The solution was filtered through a 0.45 μm microporous filter to remove a small amount of insoluble matter. The supernatant was cooled to 50 °C, and 5 mg of seed crystals of methanesulfonate form I and II of compound (I) were added. The mixture was stirred at 50 °C for 0.5 h. The temperature was then lowered to 40 °C, and 30 mL of isopropanol was slowly added dropwise over 3 h. After the addition was completed, the temperature was slowly lowered to 20 °C over 2 h. The mixture was stirred at 20 °C for 1 h and then vacuum filtered. The solid was dried under vacuum at 40 °C for 4 h to obtain a white solid, which was methanesulfonate form III. Its X-ray powder diffraction pattern is shown in Figure 24.

[0324] The TGA-DSC spectrum is shown in Figure 25. Its TGA spectrum shows a weight loss of about 1.84% at 36-145℃, accompanied by an endothermic peak.

[0325] The calorimeter moisture test showed a moisture content of 2.0%.

[0326] Formula (I) compound methanesulfonate crystal form III 1 H-NMR (400MHz, DMSO-d6): δ (ppm) 11.19 (s, 1H), 9.58-9.57 (d, J = 5.2Hz, 1H), 9.35 (brs, 1H), 8.86-8.84 (dd, J = 8.0, 1.6Hz, 1H), 8.51 -8.50(d,J=4.0Hz,1H),8.33-8.32(d,J=5.6Hz,1H),7.34-7.07(m,4H),6.95-6.93(d,J=8.0Hz,0.5H),6.53-6.51(d,J=8.0Hz,0.5H) ,5.49-5.44(dd,J=12.4,5.2Hz,1H),5.34(s,0.5H),5.15(s,0.5H),5.07-5.03(m,1H),4.86-4.81(d,J=16.0Hz,1H),3.90-3.79(m, 2H),3.71(s,3H),3.69-3.46(m,5H),3.15-2.91(m,5H),2.81-2.60(m,3H),2.41-2.32(m,6H),2.16-2.01(m,5H),1.92-1.84(m,2H).

[0327] This disclosure (I) includes examples of compound bioactivity testing.

[0328] [Correction based on Rule 91, January 2026] The control molecule used in this test example is KT-474, which was prepared according to the preparation method of compound I-417 in patent WO2020113233A1, and its structure is as follows:

[0329] Test Example 1: Degradation of IRAK4 by Compounds in SU-DHL-2 and OCI-LY3 Cells

[0330] (1) Experimental Objective

[0331] Flow cytometry was used to detect the degradation level of the compound on the IRAK4 protein in the SU-DHL-2 cell line.

[0332] (2) Experimental materials

[0333] Fix Buffer I, BD Phosflow TM 557870

[0334] Perm III buffer, BD Phosflow TM 558050

[0335] LIVE / DEAD TM Fixable Violet Dead Cell Stain Kit, Thermo Fisher, L34966

[0336] Alexa Fluor 647Mouse anti-Human IRAK4 Clone L29-525(RUO), BD Phosflow TM 560315

[0337] (3) Experimental instruments

[0338] Flow cytometer, BD, Model: BD LSRFortessa

[0339] (4) Experimental methods

[0340] Step 1: The SU-DHL-2 cell line was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin antibiotics.

[0341] Step 2: On Day 1, seed 100 μL of cells into each well of a 96-well plate at a density of 2 x 10⁵. Dissolve the compound in DMSO to a final concentration of 10 mM, then serially dilute it 10 times with culture medium at a 1:5 ratio to achieve a maximum final concentration of 2 μM in the cells. Use wells containing 0.1% DMSO as a positive control. Incubate at 37°C in a 5% CO₂ incubator for 24 hours.

[0342] Step 3: After centrifugation, wash the cells with FACS buffer. Use LIVE / DEAD. TM The Fixable Blue Dead Cell Stain Kit was used to distinguish between live and dead cells. PE Mouse Anti-Human CD14 was used to stain and label the surface of monocytes, and the cells were incubated at 4°C for 30 minutes.

[0343] Step 4: After centrifugation, wash the cells with FACS buffer. Add an equal volume of Fix buffer I to the cultured cells and fix the cells at 37°C for ten minutes.

[0344] Step 5: After centrifugation, wash the cells with FACS buffer. Add 150 μL of Perm III reagent and incubate at 4°C for 30 minutes to lyse the cell membrane. After centrifugation, wash the cells with FACS buffer.

[0345] Step 6: Stain cells with Alexa Fluor 647 Mouse anti-Human IRAK4 Antibody and incubate at 4°C for 1 hour. Perform flow cytometry analysis.

[0346] (5) Data processing

[0347] Streaming data was analyzed using the Flowjo software. The data displayed included data on DC... 50 The (half-maximum degradation concentration) was calculated using Prism 8.0.2 (GraphPad).

[0348] (6) Experimental Results

[0349] The degradation activity of the compound of formula (I) against IRAK4 is shown in Table 1.

[0350] The compound of this formula (I) generally achieves a Dmax of over 50% for both SU-DHL-2 and OCI-LY3 cells, with a maximum of around 90%.

[0351] The results showed that the compound of formula (I) exhibited good degradation activity against the IRAK4 target in SU-DHL-2 and OCI-LY3 cells.

[0352] Table 1. Degradation results of IRAK4 in SU-DHL-2 and OCI-LY3 cells

[0353] Test Example 2: Degradation of IRAK4 in human PBMCs by the compound

[0354] (1) Experimental Objective

[0355] Flow cytometry was used to detect the degradation level of the compound on IRAK4 in human peripheral blood mononuclear cells (hPBMCs).

[0356] (2) Experimental materials

[0357] Fix Buffer I, BD Phosflow TM 557870

[0358] Perm III buffer, BD Phosflow TM 558050

[0359] LIVE / DEAD TM Fixable Blue Dead Cell Stain Kit, for UV excitation, Thermo Fisher, L34962

[0360] PE Mouse Anti-Human CD14, BD Pharmingen TM 555398

[0361] Alexa Fluor 647 Mouse anti-Human IRAK4 Clone L29-525(RUO), BD Phosflow TM 560315

[0362] (3) Experimental instruments

[0363] Flow cytometer, BD, Model: BD LSRFortessa

[0364] (4) Experimental methods

[0365] After thawing, the frozen hPBMCs were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% Pen+Strep, and then incubated at 37°C for 1 hour.

[0366] On day 1, 100 μL of cells were seeded in 96-well plates at a rate of 2 x 10⁵ cells per well.

[0367] The compound was dissolved in DMSO to a final concentration of 10 mM, and then diluted 10 times in culture medium at a 1:5 ratio to achieve a maximum final concentration of 2 μM in cells. Cell wells containing 0.1% DMSO were used as a positive control.

[0368] Incubate at 37°C in an incubator containing 5% CO2 for 24 hours.

[0369] After centrifugation, the cells were washed with FACS buffer.

[0370] Use LIVE / DEAD TM The Fixable Blue Dead Cell Stain Kit was used to distinguish between live and dead cells. PE Mouse Anti-Human CD14 was used to stain and label the surface of monocytes, and the cells were incubated at 4°C for 30 minutes.

[0371] After centrifugation, the cells were washed with FACS buffer.

[0372] Add Fix buffer I in an amount equal to the volume of cultured cells and fix the cells at 37°C for ten minutes.

[0373] After centrifugation, the cells were washed with FACS buffer.

[0374] Add 150 μL of Perm III reagent and incubate at 4°C for 30 minutes to permeate the cell membrane.

[0375] After centrifugation, the cells were washed with FACS buffer.

[0376] Cells were stained with Alexa Fluor 647 Mouse anti-Human IRAK4 Antibody and incubated at 4°C for 1 hour.

[0377] Flow cytometry analysis was performed using a flow cytometer.

[0378] (5) Data processing methods

[0379] Streaming data was analyzed using the Flowjo software. The data displayed included data on DC... 50 The calculations were performed using Prism 8.0.2 (GraphPad).

[0380] (6) Experimental Results

[0381] The degradation activity of the compound of formula (I) against IRAK4 is shown in Table 2.

[0382] The results showed that the compound of formula (I) exhibited good degradation activity against IRAK4 in human PBMC cells.

[0383] Table 2. Degradation results of IRAK4 in human PBMC cells

[0384] Test Example 3: Inhibition of Compound on the Induction of Inflammatory Factor Secretion in Human PBMCs

[0385] (1) Experimental Objective

[0386] The effects of PROTAC small molecules on TLR and IL-1-induced IRAK4 downstream inflammatory factors in PBMC cells were detected using the CBA (micro-sample multi-index flow cytometry) method.

[0387] [Corrected according to Rule 91 20.01.2026] (2) Experimental materials

[0388] (3) Experimental instruments

[0389] Flow cytometer, BD, Model: BD LSRFortessa

[0390] (4) Experimental methods

[0391] Step 1: Human PBMC Recovery

[0392] Step 2: Adjust the PBMC cell density to 1E6 / mL, and seed 150μL of PBMCs with 1640 intact culture medium into a 96-well plate, and incubate at 37℃ for 1 hour.

[0393] Step 3: Inoculate 50 μL of serially diluted small molecule drug (working concentration: 10000 nM, 1:3 dilution) onto a plate in a 37°C incubator for 8 hours.

[0394] Step 4: Add 10 μL of stimulant, LPS final concentration 100 ng / mL, R848 final concentration 1 μg / mL, IL-1β final concentration 100 ng / mL respectively.

[0395] Step 5: CBA assay. The concentrations of IL6 and TNF-α in the cell culture supernatant were determined using Hu IL-6CBA Flex Set A7 and Hu TNF-CBA Flex Set D9.

[0396] Step 6: Prepare standards: 2500 pg / mL, 1250 pg / mL, ... 0.1:2 serial dilutions, 10 concentrations.

[0397] Step 7: Resuspend the mixed capture beads in capture bead diluent (1:50), add cell culture supernatant (50 μL), and incubate at room temperature for 10 minutes.

[0398] Step 8: Add the detection reagent prepared with the detection diluent and incubate at room temperature in the dark for 3 hours. At the end of the incubation, wash once with washing buffer and resuspend for detection.

[0399] (5) Data processing

[0400] IL-6 and TNFα concentrations were calculated using the standard curve method. Data showed that... 50 The calculations were performed using Prism 8.0.2.

[0401] (6) Experimental Results

[0402] The results of the inhibitory effect of the compound of formula (I) on the induction of inflammatory factor secretion in human PBMCs are shown in Tables 3 and 4.

[0403] The results show that the compound of formula (I) can effectively inhibit the secretion of inflammatory factors and has potential value in treating inflammatory diseases.

[0404] Table 3. LPS, R848, and IL-1β induce TNFα secretion in human PBMC cells

[0405] Table 4. LPS, R848, and IL-1β induce IL-6 secretion in human PBMC cells

[0406] Test Example 4: Inhibition of hERG potassium channels by the compound

[0407] (1) Experimental Objective

[0408] The effects of the compound on the current of the hERG potassium channel (human Ether-a-go-go Related Gene potassium channel) were tested using a whole-cell electrophysiological manual voltage clamp method.

[0409] (2) Experimental materials

[0410] Sodium chloride (NaCl) Sigma S7653, Potassium chloride (KCl) Sigma P9333, Magnesium chloride (MgCl2) Sigma M1028

[0411] Calcium chloride (CaCl2) Sigma 21115, Glucose Sigma G7528, HEPES Sigma H3375, EGTA Sigma E3889, Sodium hydroxide (NaOH) Sinopharm 10019718, Potassium hydroxide (KOH) Sinopharm 10017018;

[0412] CHO-hERG cell line (Chinese Hamster Ovary): Chinese hamster ovary cells stably expressing the hERG channel.

[0413] Extracellular fluid formulation (mM): 140 NaCl, 5 KCl, 1 CaCl2, 1.25 MgCl2, 10 HEPES and 10 Glucose, adjusted to pH 7.4 with NaOH.

[0414] Intracellular fluid formulation (mM): 140 KCl, 1 MgCl2, 1 CaCl2, 10 EGTA and 10 HEPES, pH adjusted to 7.2 with KOH.

[0415] (3) Experimental instruments

[0416] Patch clamp amplifier (Multiclamp 700B, Axon, USA)

[0417] Digital-to-analog converter (DigiData 1440A, Axon, USA)

[0418] Inverted microscope (IX71, Olympus, Japan)

[0419] Rapid drug delivery system (RSC-200, Bio-Logic, France)

[0420] Micromanipulator (MX7600R, Syskiyou, USA)

[0421] Electrode drawing apparatus (P-97, Sutter, USA)

[0422] Glass electrode (BF150-86-10, Sutter, USA)

[0423] Vibration damping platform and shielding mesh (63-534, TMC, USA)

[0424] Data acquisition and analysis software (pClamp, Axon, USA)

[0425] CO2 incubator (HERAcell 150i, Thermo, USA)

[0426] Biosafety cabinet (MODEL 1384, Thermo, USA)

[0427] Pure water system (Milli Q, Millipore, USA)

[0428] (4) Experimental methods

[0429] (4.1) Cell culture and treatment

[0430] CHO cells stably expressing hERG were cultured in 35 mm diameter cell culture dishes at 37°C in a 5% CO2 incubator. They were passaged every 48 hours at a 1:5 ratio. The culture medium consisted of 90% F12 (Invitrogen), 10% fetal bovine serum (Gibco), 100 g / mL G418 (Invitrogen), and 100 g / mL Hygromycin B (Invitrogen). On the day of the experiment, the cell culture medium was aspirated, the cells were rinsed once with extracellular fluid, and then 0.25% Trypsin-EDTA (Invitrogen) solution was added for digestion at room temperature for 3-5 minutes. The digestion solution was aspirated, the cells were resuspended in extracellular fluid, and then transferred to experimental dishes for electrophysiological recording.

[0431] (4.2) Compound preparation

[0432] On the day of testing, the compound was prepared as a 10 mM stock solution with DMSO, then diluted to 1 mM with DMSO, and finally diluted 1000-fold with extracellular fluid to obtain the final concentration to be tested. For the positive control compound cisapride, 10 μL of a 150 μM cisapride DMSO stock solution was added to 4990 L of extracellular fluid and diluted 500-fold to obtain the final concentration to be tested, 300 nM. The DMSO content in the final test concentration did not exceed 0.2%, as this concentration of DMSO had no effect on hERG potassium channels.

[0433] (4.3) Electrophysiological recording process

[0434] CHO cells stably expressing the hERG potassium channel were used to record hERG potassium channel currents at room temperature using whole-cell voltage-clamp technique. Glass microelectrodes were fabricated from glass electrode blanks (BF150-86-10, Sutter) using a stretching device. The tip resistance after perfusion with electrode fluid was approximately 2-5 MΩ. The glass microelectrodes were inserted into the amplifier probe to connect to the patch-clamp amplifier. Clamping voltage and data recording were controlled and recorded by computer using pClamp software, with a sampling frequency of 10 kHz and a filtering frequency of 2 kHz. After obtaining whole-cell recordings, the cells were clamped at -100 mV. The step voltage to induce hERG potassium current (IhERG) was applied from -100 mV with a 2-second depolarization voltage to +20 mV, then repolarized to -50 mV, held for 1 second, and then returned to -100 mV. This voltage stimulation was applied every 5 seconds. After confirming that the hERG potassium current was stable (1 minute), the drug delivery process began. For each test concentration of the compound, administer for at least 1 minute to reach steady state or for a maximum of 3 minutes, and test at least 2 cells for each concentration (n≥2).

[0435] (5) Data processing

[0436] Data analysis and processing were performed using pClamp, GraphPad Prism 8, and Excel software. The degree of inhibition of hERG potassium current (the peak hERG tail current induced at -50mV) by different compound concentrations was calculated using the following formula:

[0437] Inhibition%=[1–(I / Io)]×100%

[0438] Where Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current before and after drug administration, respectively.

[0439] (6) Experimental Results

[0440] Table 5. Inhibition results of hERG potassium channels

[0441] Note: @1uM refers to the concentration of the test compound.

[0442] Experimental results show that the compound of formula (I) has no significant inhibitory activity on hERG, indicating that the compound of formula (I) has good safety.

[0443] Test Example 5: In vivo pharmacokinetic determination of the compound

[0444] 1.1 Test Drugs:

[0445] This disclosure pertains to compound (I).

[0446] 1.2 Experimental Animals

[0447] Healthy male Beagle dogs, weighing approximately 8-10 kg, 3 dogs per group.

[0448] 1.3 Experimental Design:

[0449] Three Beagle dogs per group were administered the compound at a dose of 2.5 mg / kg via gavage, in a volume of 5 mL / kg, prepared with 10% DMSO + 50% PEG400 + 40% water (w / v). They were allowed free access to food and water.

[0450] Venous blood samples of 0.5-0.8 mL were collected at 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after oral administration. The samples were placed in K2EDTA tubes and centrifuged at 4 °C to separate the plasma. The concentration of compounds in the plasma was determined by liquid chromatography-tandem mass spectrometry.

[0451] 1.4 Experimental Results:

[0452] The main pharmacokinetic parameters were calculated using WinNonlin. The pharmacokinetic parameters of the canine oral drug are shown in Table 6 below. The experimental results show that the compound of formula (I) has good pharmacokinetic properties.

[0453] Table 6. Pharmacokinetic parameters of some compounds disclosed herein orally in dogs

[0454] Test Example 6: Efficacy evaluation of the compound against IL-33-induced acute inflammation in mice

[0455] (1) Experimental Objective

[0456] IL-33 is a member of the IL-1 cytokine family and a ligand for IL-1 receptor-like 1 or ST2. IL-33 transgenic mice specifically express IL-33 in keratinocytes and spontaneously develop AD (atopic dermatitis)-like eczema, suggesting that the IL-33 pathway plays a crucial role in AD. This study investigated the effects of compounds on inflammation along the IL-1R signaling pathway using an acute inflammatory mouse model induced by intraperitoneal injection of IL-33.

[0457] (2) Experimental materials and methods

[0458] Experimental animals: SPF-grade 6-8 week old male C57BL / 6J mice.

[0459] Compound preparation: 10% DMSO + 50% PEG400 + 40% sterile water for injection. It was prepared by dissolving the compound in DMSO, followed by the addition of PEG400 and sterile water for injection, and adjusting the solution to clear with 6N hydrochloric acid.

[0460] Preparation of IL-33 inducer: Prepare a 1 μg / μL stock solution of recombinant IL-33 powder from mice. When using, add 3960 μL of PBS to 40 μL of the 1 μg / μL IL-33 solution, invert and mix well to obtain 4 mL of IL-33 solution with a concentration of 1 μg / 100 μL. Prepare and use immediately.

[0461] Compound administration and IL-33 inflammation induction: Mice were randomly divided into groups according to body weight and orally administered the compound at final doses of 15 mg / kg, 50 mg / kg, and 150 mg / kg twice daily. Four hours after the last administration of the compound, mice were intraperitoneally injected with IL-33 1 μg / 100 μL PBS to induce inflammation.

[0462] Termination, Sample Collection and Data: Plasma, peritoneal lavage fluid and spleen were collected for subsequent sample indicator testing.

[0463] Eight hours after the last administration, the experiment was terminated. Blood was collected from the heart of anesthetized mice. After anticoagulation with EDTA-K2, the blood samples were centrifuged at 3500 rpm for 15 min, and the supernatant plasma was collected. Euthanized mice were then disinfected by immersion in 75% alcohol for 3 min in a sterile laminar flow hood. 2.5 mL of pre-cooled PBS (4°C) was injected intraperitoneally, and the abdomen was massaged for 5 min. The abdominal skin was then cut open with sterile scissors and forceps to expose the peritoneum. The sternum was lifted at the xiphoid process using sterile forceps, and a 1 mm incision was made in the peritoneum at the xiphoid process. 2 A small incision was made, and a pipette was inserted into the peritoneal cavity to gently blow and collect 2 mL of the lavage fluid. The fluid was then aliquoted into 1.5 mL centrifuge tubes and stored at -80°C. The expression levels of inflammatory factors in mouse plasma and peritoneal lavage fluid were detected by ELISA, and the expression level of IRAK4 protein in mouse spleen was detected by LC-MS.

[0464] (3) Experimental Results and Analysis

[0465] Table 7. Endpoint plasma and tissue drug concentrations

[0466] Table 7 shows that the concentrations of compound (I) in plasma and spleen at a dose of 50 mpk were the same as those of KT-474 at 150 mpk, indicating that compound (I) requires a lower dosage.

[0467] Figures 26-29 show the concentration-dependent decrease of inflammatory factors in mouse plasma and peritoneal lavage fluid after three PO doses of compound (I). At comparable plasma drug concentrations, compound (I) exhibited superior anti-inflammatory effects compared to KT474.

[0468] Figure 30 shows that, under comparable drug concentrations in the spleen, compound (I) exhibits 2-3 times better degradation performance at 50 mpk than KT-474 at 150 mpk.

[0469] This disclosure (I) includes the testing of the crystal form, salt, and crystal form effect of the compound.

[0470] [Corrected according to Rule 91, 2001.2026] Test Example 7 Hygroscopicity (DVS)

[0471] The hygroscopicity of the samples was determined using a DVS dynamic moisture adsorption instrument. Appropriate amounts of crystal form A and p-toluenesulfonate crystal form I were placed in a sample basket. After reaching equilibrium at 0% humidity, the hygroscopicity of the samples was measured during one cycle from 0% RH to 90% RH (Cycle 2sorp) and from 90% RH back to 0% RH.

[0472] Table 8. Hygroscopicity of the crystal form of compound (I)

[0473] According to the experimental results, the hygroscopicity of compound (I) is significantly reduced after salt formation. The hygroscopicity of p-toluenesulfonate crystal form I is better than that of free base crystal form A. Its DVS spectra are shown in Figure 3 and Figure 18, respectively.

[0474] [Corrected according to Rule 91, 2001.2026] Test Example 8: Influence Factor Experiment

[0475] The influencing factor test was conducted according to the "Guidelines for Stability Testing of Active Pharmaceutical Ingredients and Preparations" (Chinese Pharmacopoeia 2015 Edition, 9001), examining the stability of samples with different crystal forms under high temperature, high humidity, and light exposure. Under high temperature conditions, samples were double-packaged in polyethylene bags and aluminum foil bags and placed in constant temperature chambers at 40℃ and 60℃. Under high humidity conditions, samples were exposed and placed in constant humidity chambers at 75% RH and 25℃ and 90% RH and 25℃. Under light exposure conditions, samples were exposed and spread into a thin layer and placed in a light chamber with a light intensity ≥5000 lx and a near-ultraviolet lamp energy of not less than 200 W·hr / m². 2 .

[0476] The experimental results are shown in the table below.

[0477] Table 9.1 Influencing Factors of Free Alkali Crystal Form A (Experimental)

[0478] Table 9.2 Influencing Factors of Toluenesulfonate Crystal Form I Experiment

[0479] Table 9.3 Influencing Factors of Methanesulfonate Crystal Form III (Test)

[0480] [Corrected according to Rule 91, 20.01.2026] The results show that under high temperature, high humidity and light conditions, the crystal forms of free crystal form A, p-toluenesulfonate crystal form I and methanesulfonate crystal form III did not change before and after the stability test, and the purity did not change much. Therefore, free crystal form A, p-toluenesulfonate crystal form I and methanesulfonate crystal form III have good physicochemical stability.

[0481] [Corrected according to Rule 91, 2001.2026] Test Example 9: Pharmacokinetic Study

[0482] (1) Solubility of the crystal form of compound (I) in the solvent

[0483] This study investigates the differences in solubility of the compound of the present invention in in vitro solvent systems for p-toluenesulfonate crystal form I, methanesulfonate crystal form III, and free base crystal form A.

[0484] 1. Test materials

[0485] 1.1 Test Drugs

[0486] The p-toluenesulfonate crystal form I, the methanesulfonate crystal form III, and the free base crystal form A of the compound of formula (I).

[0487] 1.2 Main Instruments

[0488] Liquid chromatograph (Arc, Waters), magnetic stirrer (RCT basic, IKA), electronic balance (XSE205, Mettler).

[0489] 1.3 Solvent

[0490] [Corrected according to Rule 91, January 2026] Vitamin E polyethylene glycol succinate (TPGS, Shanghai Lingkai Pharmaceutical Technology Co., Ltd.)

[0491] 2. Steps and methods

[0492] 2.1 Sample preparation and pretreatment

[0493] [Corrected according to Rule 91, 20.01.2026] Sample preparation: Weigh appropriate amounts of p-toluenesulfonate crystal form I and methanesulfonate crystal form III respectively, add them to a 10% TPGS solvent system to obtain a test sample of 60 mg / ml, and stir to obtain a homogeneous suspension. Weigh an appropriate amount of free alkali crystal form A, add 10% TPGS to obtain a test sample of 60 mg / ml, and stir to obtain a homogeneous suspension.

[0494] Sample pretreatment: The prepared test suspension was filtered through a 0.22 μm organic filter membrane, and the filtrate was diluted with 50% acetonitrile aqueous solution. The diluted test suspension was quantitatively analyzed using the standard curve method.

[0495] 2.2 Liquid Phase Analysis

[0496] Mobile phase: A is 0.1% phosphoric acid aqueous solution; B is acetonitrile;

[0497] Chromatographic column: Waters Xbridge BEH C18 (4.6×150mm, 3.5μm);

[0498] Flow rate: 1.0 ml / min;

[0499] Wavelength: 230 nm;

[0500] Gradient elution: Gradient elution for 0 - 10 minutes.

[0501] 3. Test Results and Analysis

[0502] The solubility of different crystal forms of the compound in vitro solvent is shown in Table 10 below.

[0503] Table 10 Determination Results of in vitro Solubility of Different Crystal Forms of Compound (I)

[0504] (2) Comparative Study on the Pharmacokinetics of Crystal Forms of Compound (I) in Wistar Rats

[0505] Using Wistar rats as the test animals, the pharmacokinetic behavior of the compound of the present invention in the plasma of rats after oral administration at a dose of 500 mg / kg was studied.

[0506] 1. Test Scheme

[0507] 1.1 Test drugs:

[0508] p-Toluenesulfonate Crystal Form I, Methanesulfonate Crystal Form III, Free Base Crystal Form A.

[0509] 1.2 Test animals

[0510] 16 Wistar rats (4 rats / group), male, Beijing Vital River Laboratory Animal Technology Co., Ltd., Animal Production License Number (SCXK(Beijing)2021 - 0006).

[0511] 1.3 Administration:

[0512] 16 Wistar rats, male; after free feeding, p.o., the dose of PO administration is 500 mg / kg, and the administration volume is 10 mL / kg.

[0513] 1.4 Experimental equipment

[0514] The high-speed refrigerated centrifuge was purchased from Sigma, the vortex mixer from DLAB, the -80℃ medical cryopreservation box from Qingdao Haier Biomedical Co., Ltd., the medical refrigerator from Qingdao Haier Biomedical Co., Ltd., and the analytical balance from Jinan Taihe Yingke Co., Ltd.

[0515] 1.5 Sample Collection

[0516] In Wistar rats, 0.2 mL of blood was collected from the jugular vein at 0 hours before administration and at 1, 2, 4, 8, 12, 24 and 48 hours after administration. The blood was placed in a heparin sodium test tube, centrifuged at 6000 rpm for 5 min at 4°C, and the plasma was separated and stored at -80°C.

[0517] 1.6 Sample Preparation

[0518] 1) Add 200 μL of acetonitrile (containing internal standard aprepitant) to 20 μL of plasma sample to precipitate, mix and centrifuge for 10 minutes.

[0519] 2) Take 80 μL of the supernatant solution after treatment and add it to 80 μL of ultrapure water containing 0.1% formic acid. Vortex mix, centrifuge, and take the supernatant for LC / MS / MS analysis to determine the concentration of the analyte.

[0520] 1.7 Liquid chromatography-mass spectrometry

[0521] Liquid phase conditions: Shimadzu Exion LC

[0522] Mass spectrometry conditions: Mass spectrometer

[0523] Chromatographic column: Waters Xbrideg protein BEH C4 (3.5 μm, 2.1 × 50 mm)

[0524] Mobile phase: Solution A is 0.1%-0.2% formic acid in water, Solution B is 0.2% formic acid in acetonitrile

[0525] Flow rate: 0.9 mL / min

[0526] Elution time: gradient elution 0-3.0 minutes.

[0527] 2. Experimental Results and Analysis

[0528] The main pharmacokinetic parameters were calculated using WinNonlin 8.3.5. The pharmacokinetic parameters of the orally administered drug in rats are shown in Table 11 below:

[0529] Table 11. Pharmacokinetic parameters of some compounds of the present invention orally administered to rats.

[0530] Experimental results show that the compound of formula (I) of this invention exhibits superior pharmacokinetic properties in crystal form I of toluenesulfonate compared to other crystal forms.

Claims

1. A compound of the following formula (I), crystal form A, with the chemical name: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-(7-(4-(1-(2,6-dioxopiperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)but-3-yn-1-yl)-7-azaspiro[3.5]nonane-2-yl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-amide, Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.48, 10.34, 10.98 and 14.19, with a 2θ error range of ±0.2°; Preferably, the crystal form A, when subjected to Cu-Kα radiation, has a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 5.31, 9.22, 10.76, 16.17, 18.30, 22.15, and 25.36, with a 2θ error range of ±0.2°. Preferably, the crystal form A, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 11.76, 12.67, 13.32, 14.63, 15.11, 15.76, 16.89, 17.82, 18.76, 19.75, 20.52, 21.35, 22.87, 23.45, and 26.50, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of crystal form A are shown in Table 1a below: Table 1a shows the XRPD diffraction peak analysis data for crystal form A of compound (I). Preferably, the crystal form A, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is basically as shown in Figure 1.

2. The compound crystal form A represented by formula (I) as described in claim 1, characterized in that... Its DSC spectrum shows an endothermic peak between 180℃ and 220℃; preferably, an endothermic peak is found at 204.8℃ ± 2℃; preferably, the TGA-DSC spectrum of the crystal form A is basically as shown in Figure 2.

3. A crystal form B of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.87, 10.82 and 11.52, with a 2θ error range of ±0.2°; Preferably, the crystal form B, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 13.58, 16.45, 17.00, 18.86, 19.76, 20.43, 22.64, 23.76, and 27.88, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of crystal form B are shown in Table 2a below: Table 2a shows the XRPD diffraction peak analysis data for crystal form B of compound (I). Preferably, the crystal form B, when subjected to Cu-Kα radiation, exhibits an X-ray powder diffraction pattern as shown in Figure 4.

4. The crystal form B of the compound of formula (I) as described in claim 3, characterized in that, Its DSC spectrum shows an endothermic peak between 170℃ and 210℃; preferably, an endothermic peak is found at 196.6℃ ± 2℃; its TGA spectrum shows a weight loss of approximately 3.8% between 35℃ and 209℃. Preferably, the TGA-DSC spectrum of crystal form B is basically as shown in Figure 5.

5. A crystal form C of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.85, 8.80, 10.82, 11.46 and 13.54, with a 2θ error range of ±0.2°; Preferably, the crystal form C, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 9.72, 12.80, 16.50, 16.87, 17.67, 18.87, 19.28, 19.80, 20.56, 22.65, 23.84, and 27.95, with a 2θ error range of ±0.2°. Preferably, the crystal form C, when subjected to Cu-Kα radiation, has a characteristic peak in its X-ray powder diffraction pattern at one or more of the 2θ values ​​of 22.10, 23.21, and 24.36, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of crystal form C are shown in Table 3a below: Table 3a shows the XRPD diffraction peak analysis data for crystal form C of compound (I). Preferably, the X-ray powder diffraction pattern of the crystal form C, obtained using Cu-Kα radiation, is basically as shown in Figure 6.

6. The crystal form C of the compound of formula (I) as described in claim 5, characterized in that, Its DSC spectrum shows an endothermic peak between 170℃ and 220℃; preferably, an endothermic peak is found at 195.7℃ ± 2℃; its TGA spectrum shows a weight loss of approximately 5.7% between 36℃ and 213℃. Preferably, the TGA-DSC spectrum of the crystal form C is basically as shown in Figure 7.

7. A crystal form D of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.81, 8.81, 9.76, 10.89, 11.37, and 13.60, with a 2θ error range of ±0.2°; Preferably, the crystal form D, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 3.92, 12.78, 16.60, 16.89, 17.67, 18.78, 19.26, 19.84, 20.41, 20.69, 22.06, 22.63, 23.91, and 28.10, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of the crystal form D are shown in Table 4a below: Table 4a shows the XRPD diffraction peak analysis data for crystal form D of compound (I). Preferably, the crystal form D, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is basically as shown in Figure 8.

8. The crystal form D of the compound of formula (I) as described in claim 7, characterized in that, Its DSC spectrum shows an endothermic peak between 170℃ and 220℃; preferably, an endothermic peak is observed at 202.35℃ ± 2℃; its TGA spectrum shows a weight loss of approximately 7.1% between 36℃ and 217℃. Preferably, the TGA-DSC spectrum of the crystal form D is basically as shown in Figure 9.

9. A crystal form E of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.78, 9.74, 10.76, 11.48 and 13.46, with a 2θ error range of ±0.2°; Preferably, the crystal form E, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 15.15, 16.54, 16.84, 17.56, 18.82, 19.28, 19.80, 20.30, 22.60, 23.80, and 27.80, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of the crystal form E are shown in Table 5a below: Table 5a shows the XRPD diffraction peak analysis data for crystal form E of compound E. Preferably, the crystal form E, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is basically as shown in Figure 10.

10. The crystal form E of the compound of formula (I) as described in claim 9, characterized in that, Its DSC spectrum shows an endothermic peak between 170℃ and 220℃; preferably, an endothermic peak is observed at 197.72℃ ± 2℃; its TGA spectrum shows a weight loss of approximately 5.2% between 95℃ and 214℃. Preferably, the TGA-DSC spectrum of the crystal form E is basically as shown in Figure 11.

11. A crystal form F of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.93, 8.93, 11.04, 11.59 and 13.82, with a 2θ error range of ±0.2°; Preferably, the crystal form F, when subjected to Cu-Kα radiation, exhibits a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 4.00, 9.67, 12.87, 16.37, 17.02, 19.82, 20.71, 21.06, 24.17, and 28.34, with a 2θ error range of ±0.2°. Preferably, the crystal form F, when subjected to Cu-Kα radiation, has a characteristic peak in its X-ray powder diffraction pattern at one or more of the following 2θ values: 18.04, 18.52, 19.04, 19.45, 22.39, and 22.93, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of the crystal form F are shown in Table 6a below: Table 6a shows the XRPD diffraction peak analysis data for crystal form F of compound (I). Preferably, the crystal form F, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is basically as shown in Figure 12.

12. The crystal form F of the compound of formula (I) as described in claim 11, characterized in that, Its DSC spectrum shows an endothermic peak between 170℃ and 220℃; preferably, an endothermic peak is observed at 203.24℃ ± 2℃; its TGA spectrum shows a weight loss of approximately 4.4% between 36℃ and 215℃. Preferably, the TGA-DSC spectrum of the crystal form F is basically as shown in Figure 13.

13. A crystal form G of a compound of formula (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 6.11, 6.54, 7.91, 8.93, 9.37 and 11.06, with a 2θ error range of ±0.2°; Preferably, the crystal form G, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 4.00, 11.56, 13.82, 16.41, 17.04, 18.02, 18.91 and 19.84, with a 2θ error range of ±0.2°. Preferably, the crystal form G, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern with one or more characteristic peaks at 2θ values ​​of 12.91, 15.02, 16.04, 20.71, 20.98, 22.84, 24.19, 25.71, and 28.30, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of the crystal form G are shown in Table 7a below: Table 7a shows the XRPD diffraction peak analysis data for crystal form G of compound G. Preferably, the crystal form G, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is basically as shown in Figure 14.

14. A pharmaceutically acceptable salt formed by a compound of formula (I) and one or more acid molecules, Its features are, The acid is selected from organic acids or inorganic acids.

15. The pharmaceutically acceptable salt of the compound of formula (I) as described in claim 14 is selected from hydrochloride, maleate, p-toluenesulfonate, methanesulfonate, hydrobromide, succinate, L-malate, L-tartrate, benzenesulfonate, phosphate, gentianate, oxalate, fumarate and citrate. Preferably, the pharmaceutically acceptable salt of the compound of formula (I) is p-toluenesulfonate or methanesulfonate; Preferably, the pharmaceutically acceptable salt of the compound of formula (I) is a toluenesulfonate or a methanesulfonate.

16. A compound of formula (I) p-toluenesulfonate, crystal form I, Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 5.37, 7.41, 8.76, 9.91 and 14.24, with a 2θ error range of ±0.2°.

17. The p-toluenesulfonate crystal form I as described in claim 16, characterized in that, Using Cu-Kα radiation, its X-ray powder diffraction pattern also shows characteristic peaks at one or more of the following 2θ values: 10.86, 11.74, 14.89, 15.21, 16.43, 18.28, 21.39, and 21.67, with a 2θ error range of ±0.2°.

18. The p-toluenesulfonate crystal form I as described in claim 16 or 17, characterized in that, The compound of formula (I) p-toluenesulfonate crystal form I, when subjected to Cu-Kα radiation, exhibits characteristic peaks in its X-ray powder diffraction pattern at one or more of the following 2θ values: 12.61, 12.91, 19.43, 20.37, 22.54, 23.26, 24.21, 24.69, 25.43, 25.93, 26.84, 27.73, and 29.17, with a 2θ error range of ±0.2°. Preferably, the X-ray powder diffraction pattern data of the p-toluenesulfonate crystal form I of the compound of formula (I) are shown in Table 8a below: Table 8a shows the XRPD diffraction peak analysis data for p-toluenesulfonate crystal form I of compound (I).

19. The p-toluenesulfonate crystal form I as described in claim 16, characterized in that, The X-ray powder diffraction pattern of the compound of formula (I) p-toluenesulfonate, crystal form I, using Cu-Kα radiation, is basically shown in Figure 16.

20. The p-toluenesulfonate crystal form I of compound (I) as described in claim 16, characterized in that, Its DSC spectrum shows an endothermic peak at 50℃-130℃, preferably at 95.2℃±2℃; an endothermic peak at 220℃-250℃, preferably at 237.7℃±2℃; its TGA spectrum shows a weight loss of about 2.1% at 35~134℃. Preferably, the TGA-DSC spectrum of the p-toluenesulfonate of compound (I) is basically as shown in Figure 17.

21. The p-toluenesulfonate crystal form I of compound (I) as described in claim 16, characterized in that, The molar ratio of compound (I) to p-toluenesulfonic acid is 1:

1.

22. The p-toluenesulfonate crystal form I of compound (I) as described in claim 16, characterized in that, The crystal form is a hydrate.

23. The p-toluenesulfonate crystal form I of compound (I) as described in claim 16, characterized in that, The crystal form is a monohydrate.

24. The p-toluenesulfonate crystal form I of compound (I) as described in claim 16, characterized in that... Crystal form I, determined by Microcrystalline Electron Diffraction (MicroED), has a space group of P21 and a unit cell parameter of [missing information]. α=90°, β=91.1(4)°, γ=90°, The smallest asymmetric unit contains two molecules of compound (I), two p-toluenesulfonic acid molecules, and two water molecules, and each unit cell contains two smallest asymmetric units.

25. A compound of formula (I) p-toluenesulfonate crystal form II, Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.35, 8.74, 9.84, 11.69, 14.65 and 18.19, with a 2θ error range of ±0.2°; Preferably, the p-toluenesulfonate crystal form II of the compound of formula (I) is prepared using Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 12.54, 16.43 and 20.60, with an error range of ±0.2°; Preferably, the X-ray powder diffraction pattern data of the p-toluenesulfonate crystal form II of compound (I) are shown in Table 9a below: Table 9a shows the XRPD diffraction peak analysis data for compound p-toluenesulfonate crystal form II of formula (I). Preferably, the X-ray powder diffraction pattern of the compound of formula (I) p-toluenesulfonate crystal form II, when subjected to Cu-Kα radiation, is basically as shown in Figure 22.

26. The p-toluenesulfonate crystal form II of compound (I) as described in claim 25, characterized in that, Its DSC spectrum shows an endothermic peak at 70℃-130℃, preferably at 100.14℃±2℃; an endothermic peak at 220℃-250℃, preferably at 239.22℃±2℃; its TGA spectrum shows a weight loss of about 2.3% at 36~169℃. Preferably, the TGA-DSC spectrum of the p-toluenesulfonate crystal form II is basically as shown in Figure 23.

27. The p-toluenesulfonate crystal form II of compound (I) as described in claim 25 or 26, characterized in that, The molar ratio of compound (I) to p-toluenesulfonic acid is 1:

1.

28. The p-toluenesulfonate crystal form II of compound (I) as described in claim 27, characterized in that, The crystal form is a hydrate; Preferably, the compound of formula (I) is p-toluenesulfonate crystal form II, wherein the crystal form is a monohydrate.

29. A methanesulfonate crystal form III of compound (I), Its features are, Using Cu-K α The radiation, its X-ray powder diffraction pattern has characteristic peaks at 2θ values ​​of 7.87, 9.41, 11.80, 12.13, 13.65, 15.33 and 15.86, with a 2θ error range of ±0.2°; Preferably, the methanesulfonate crystal form III of the compound of formula (I) uses Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 8.72, 9.15, 17.06, 17.71, 18.04, 19.26, 22.49 and 23.69, with a 2θ error range of ±0.2°; Preferably, the methanesulfonate crystal form III of the compound of formula (I) uses Cu-K α The radiation, its X-ray powder diffraction pattern also has one or more characteristic peaks at 2θ values ​​of 12.67, 18.39, 19.78, 20.21, 21.02, 24.54, 25.76 and 26.50, with a 2θ error range of ±0.2°; Preferably, the X-ray powder diffraction pattern data of the methanesulfonate crystal form III of the compound of formula (I) are shown in Table 10a below: Table 10a XRPD diffraction peak analysis data of methanesulfonate crystal form III of compound (I) Preferably, the methanesulfonate crystal form III of the compound of formula (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is basically as shown in Figure 24.

30. The methanesulfonate crystal form III of compound (I) as described in claim 29, characterized in that, Its DSC spectrum shows an endothermic peak at 70℃-130℃, preferably at 96.77℃±2℃; its TGA spectrum shows a weight loss of 1.84% at 36~145℃. Preferably, the TGA-DSC spectrum of the crystal form III is basically as shown in Figure 25.

31. The methanesulfonate crystal form III of compound (I) as described in claim 29 or 30, characterized in that, The molar ratio of compound (I) to p-toluenesulfonic acid is 1:

1.

32. The methanesulfonate crystal form III of compound (I) as described in claim 31, characterized in that, The crystal form is a hydrate; Preferably, the methanesulfonate of formula (I) is crystal form III, wherein the crystal form is a monohydrate.

33. A pharmaceutical formulation comprising the crystal form according to any one of claims 1-13, 16-32, or the salt according to any one of claims 14-15, and any one or more pharmaceutically acceptable carriers.

34. Use of a crystal form according to any one of claims 1-13, 16-32, or a salt according to any one of claims 14-15 in the preparation of a medicament for the prevention and / or treatment of IRAK4-mediated diseases; Preferably, the IRAK4-mediated related diseases are selected from immune inflammatory diseases; Preferably, the IRAK4-mediated related diseases are selected from hidradenitis suppurativa, rheumatoid arthritis, atopic dermatitis, lupus erythematosus, gouty arthritis, psoriasis, asthma, chronic obstructive pulmonary disease, polyposis sinusitis, inflammatory bowel disease, respiratory distress syndrome, chronic kidney disease, acute lung injury, IgA nephropathy, gout, multiple sclerosis, and ischemic stroke.

35. A method for preparing methanesulfonate crystal form I of compound (I) as described in any one of claims 16-24, characterized in that, Includes the following steps: (1) The compound of formula (I) and p-toluenesulfonic acid are mixed in a mixed solvent of organic solvent and water and heated to dissolve; (2) Cool down to saturation of the solution, and optionally add p-toluenesulfonate crystal form I of compound (I) as a seed crystal; (3) Continue cooling or optionally add an antisolvent for crystallization; (4) Solid separation and drying yielded p-toluenesulfonate crystal form I of compound (I).

36. [Correction 20.01.2026 according to Rule 91] The preparation method according to claim 35, characterized in that, The reaction conditions in the preparation method further include one or more of the following: (a) The molar ratio of p-toluenesulfonic acid to the compound of formula (I) is 1.0 to 1.5:1, preferably 1.0 to 1.2:1; (b) Cooling to solution saturation is preferably done at about 40-55°C; (c) Continue cooling, preferably to a temperature not lower than 0°C; (d) The organic solvent refers to an organic solvent miscible with water, selected from alcohols, nitriles, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile is selected from acetonitrile; and the ketone is selected from acetone. (e) The antisolvent is selected from alcohols, nitriles, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile is selected from acetonitrile; and the ketone is selected from acetone. (f) The amount of seed crystals added can be 0.1 wt% to 10 wt% of the amount of the compound of formula (I), preferably 0.5 wt% to 5 wt%.

37. [Correction 20.01.2026 according to Rule 91] The preparation method according to claim 36, characterized in that, The seed crystals are prepared by the following method: The compound of formula (I) and p-toluenesulfonic acid were mixed in a solvent, slurried at room temperature, the solid was collected and dried to obtain p-toluenesulfonate crystal form I of compound (I); Preferably, the solvent is selected from organic solvents or mixtures of organic solvents and water, wherein the organic solvent is selected from alcohols, nitrile solvents, and ketones; wherein the alcohol is selected from ethanol, isopropanol, and n-propanol; the nitrile solvent is selected from acetonitrile; and the ketone solvent is selected from acetone. Preferably, the mixing time for pulping is selected from 1 hour to 96 hours; Preferably, the molar ratio of p-toluenesulfonic acid to the compound of formula (I) is 1.0 to 1.5:1, more preferably 1.0 to 1.2:1.

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