Crystal form, salt form, composition and preparation method of pyridazine compound

Abstract

The invention discloses a crystal form, a salt form, a composition and a preparation method of a pyridazine compound, and particularly discloses a preparation method and application of a crystal form A and a composition of a compound shown as a formula (I), a crystal form B of a compound shown as a formula (II) and a crystal form C of a compound shown as a formula (III).

Description

Crystal form, salt form, composition and preparation method of pyridazine compound

[0001] This application claims priority to:

[0002] CN202210918680.X, August 1, 2022;

[0003] CN202210940251.2, August 5, 2022. Technical Field

[0004] The present invention relates to a crystal form, a salt form, a composition and a preparation method of a pyridazine compound, and specifically discloses a crystal form A, a salt form and a composition of a compound of formula (I), a crystal form B of a compound of formula (II) and a crystal form C of a compound of formula (III) and applications thereof. Background Art

[0005] The NLRP3 inflammasome is a multiprotein complex that plays a crucial role in the pathogenesis of innate immunity and inflammatory diseases. The NLRP3 inflammasome is composed of NOD-like receptors (NLRs), apoptosis-associated speck-like protein containing a CARD (ASC), and caspase-1. Exogenous pathogens or endogenous risk factors such as mitochondrial reactive oxygen species, oxidized mitochondrial DNA, β-amyloid protein, or α-synuclein can activate NLRP3. Activated NLRP3, ASC, and caspase-1 form the activated NLRP3 inflammasome. Caspase-1 further hydrolyzes pro-IL-1β and pro-IL-18, leading to the release of the active cytokines IL-1β and IL-18. The secretion of these cytokines can lead to pyroptosis or neuronal damage. NLRP3 inflammasome plays an important role in the occurrence of various autoimmune diseases, cardiovascular diseases, neurodegenerative diseases and tumors (Nature Reviews Drug Discovery, 2018, 17(8): 588-606.).

[0006] Currently, no NLRP3 inhibitors are commercially available. The preclinical compound MCC950 has shown significant inhibitory activity against NLRP3 (Sci. Transl. Med. 10, eaah4066 (2018)). Drugs such as OLT-1177, Inzomelid, Selnoflast, and IFM-2427 are currently in clinical development. The development of NLRP3 inhibitors holds broad potential for application.

[0007] Summary of the Invention

[0008] The present invention also provides a crystalline form A of the compound of formula (I), which has an X-ray powder diffraction pattern (XRPD) having characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 16.53±0.20°, and 18.12±0.20°;

[0009] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 13.48±0.20°, 16.53±0.20°, 18.12±0.20°, 21.44±0.20° and 24.06±0.20°.

[0010] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 12.72±0.20°, 13.48±0.20°, 16.53±0.20°, 18.12±0.20°, 21.44±0.20°, 24.06±0.20° and 25.55±0.20°.

[0011] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.20°, 8.41±0.20°, 12.72±0.20°, 13.48±0.20°, 14.72±0.20°, 15.92±0.20°, 16.53±0.20°, 16.89±0.20°, 18.12±0.20°, 21.44±0.20°, 24.06±0.20°, 25.55±0.20° and 27.12±0.20°.

[0012] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.10°, 8.41±0.10°, 12.72±0.10°, 13.48±0.10°, 14.72±0.10°, 15.92±0.10°, 16.53±0.10°, 16.89±0.10°, 18.12±0.10°, 21.44±0.10°, 24.06±0.10°, 25.55±0.10° and 27.12±0.10°.

[0013] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.20°, 8.41±0.20°, 12.16±0.20°, 12.72±0.20°, 13.48±0.20°, 14.72±0.20°, 15.92±0.20°, 16.53±0.20°, 16.89±0.20°, 17.37±0.20°, 18.12±0.20°, 21.44±0.20°, 24.06±0.20°, 24.84±0.20°, 25.55±0.20°, 27.12±0.20° and 29.03±0.20°.

[0014] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.10°, 8.41±0.10°, 12.16±0.10°, 12.72±0.10°, 13.48±0.10°, 14.72±0.10°, 15.92±0.10°, 16.53±0.10°, 16.89±0.10°, 17.37±0.10°, 18.12±0.10°, 21.44±0.10°, 24.06±0.10°, 24.84±0.10°, 25.55±0.10°, 27.12±0.10° and 29.03±0.10°.

[0015] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.20°, 8.41±0.20°, 11.58±0.20°, 12.16±0.20°, 12.72±0.20°, 13.07±0.20°, 13.48±0.20°, 14.72±0.20°, 15. 92±0.20°, 16.53±0.20°, 16.89±0.20°, 17.37±0.20°, 18.12±0.20°, 18.43±0.20°, 19.11±0.20°, 21.24±0.20°, 21.44±0.20°, 22.41±0.20°, 23.02±0.20°, 23.23±0.20 °, 23.52±0.20°, 24.06±0.20°, 24.84±0.20°, 25.34±0.20°, 25.55±0.20°, 26.08±0.20°, 26.49±0.20°, 26.89±0.20°, 27.12±0.20°, 27.92±0.20°, 28.62±0.20°, 29.03 ±0.20°, 29.75±0.20°, 30.10±0.20°, 31.21±0.20°, 32.10±0.20°, 32.89±0.20°, 33.26±0.20°, 34.45±0.20°, 35.64±0.20°, 36.06±0.20°, 37.24±0.20° and 38.84±0.20°.

[0016] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80±0.10°, 8.41±0.10°, 11.58±0.10°, 12.16±0.10°, 12.72±0.10°, 13.07±0.10°, 13.48±0.10°, 14.72±0.10°, 15. 92±0.10°, 16.53±0.10°, 16.89±0.10°, 17.37±0.10°, 18.12±0.10°, 18.43±0.10°, 19.11±0.10°, 21.24±0.10°, 21.44±0.10°, 22.41±0.10°, 23.02±0.10°, 23.23±0.10 °, 23.52±0.10°, 24.06±0.10°, 24.84±0.10°, 25.34±0.10°, 25.55±0.10°, 26.08±0.10°, 26.49±0.10°, 26.89±0.10°, 27.12±0.10°, 27.92±0.10°, 28.62±0.10°, 29.03 ±0.10°, 29.75±0.10°, 30.10±0.10°, 31.21±0.10°, 32.10±0.10°, 32.89±0.10°, 33.26±0.10°, 34.45±0.10°, 35.64±0.10°, 36.06±0.10°, 37.24±0.10° and 38.84±0.10°.

[0017] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 5.80, 8.41, 11.58, 12.16, 12.72, 13.07, 13.48, 14.72, 15.92, 16.53, 16.89, 17.37, 18.12, 18.43, 19.11, 21.24, 21.44, 22.41, 2 3.02, 23.23, 23.52, 24.06, 24.84, 25.34, 25.55, 26.08, 26.49, 26.89, 27.12, 27.92, 28.62, 29.03, 29.75, 30.10, 31.21, 32.10, 32.89, 33.26, 34.45, 35.64, 36.06, 37.24, 38.84.

[0018] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 16.53±0.20°, and / or 18.12±0.20°, and / or 5.80±0.20°, and / or 11.58±0.20°, and / or 12.16±0.20°, and / or 12.72±0.20°, and / or 13.07±0.20°, and / or 13.48±0.20°, and / or 14.72±0.20°, and / or 15.92±0.20°, and / or 16.89±0.20°, and / or 17.37±0.20°, and / or 18. 43±0.20°, and / or 19.11±0.20°, and / or 21.24±0.20°, and / or 21.44±0.20°, and / or 22.41±0.20°, and / or 23.02±0.20°, and / or 23.23±0.20°, and / or 23.52±0.20°, and / or 24.06±0.20°, and / or 24.84±0.20°, and / or 25.34±0.20°, and / or 25.55±0.20°, and / or 26.08±0.20°, and / or 26.49±0.20°, and / or 26.89±0.20°, and / or 27.12±0.20°, and / or 27.92±0.20°, and / or 28.62±0.20°, and / or 29.03±0.20°, and / or 29.75±0.20°, and / or 30.10±0.20°, and / or 31.21±0.20°, and / or 32.10±0.20°, and / or 32.89±0.20°, and / or 33.26±0.20°, and / or 34.45±0.20°, and / or 35.64±0.20°, and / or 36.06±0.20°, and / or 37.24±0.20°, and / or 38.84±0.20°.

[0019] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form A has characteristic diffraction peaks at the following 2θ angles: 8.41±0.10°, 16.53±0.10°, and / or 18.12±0.10°, and / or 5.80±0.10°, and / or 11.58±0.10°, and / or 12.16±0.10°, and / or 12.72±0.10°, and / or 13.07±0.10°, and / or 13.48±0.10°, and / or 14.72 ± 0.10°, and / or 15.92 ± 0.10°, and / or 16.89 ± 0.10°, and / or 17.37 ± 0.10°, and / or 18.43 ± 0.10°, and / or 19.11 ± 0.10°, and / or 21.24 ± 0.10°, and / or 21.44 ± 0.10°, and / or 22.41 ± 0.10°, and / or 23.02 ± 0.10°, and / or 23.23 ± 0.10°, and / or 23. 52±0.10°, and / or 24.06±0.10°, and / or 24.84±0.10°, and / or 25.34±0.10°, and / or 25.55±0.10°, and / or 26.08±0.10°, and / or 26.49±0.10°, and / or 26.89±0.10°, and / or 27.12±0.10°, and / or 27.92±0.10°, and / or 28.62±0.10°, and / or 29.03±0.10° .10°, and / or 29.75±0.10°, and / or 30.10±0.10°, and / or 31.21±0.10°, and / or 32.10±0.10°, and / or 32.89±0.10°, and / or 33.26±0.10°, and / or 34.45±0.10°, and / or 35.64±0.10°, and / or 36.06±0.10°, and / or 37.24±0.10°, and / or 38.84±0.10°.

[0020] In some embodiments of the present invention, the XRPD pattern of the above-mentioned crystal form A is basically as shown in Figure 1.

[0021] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form A is shown in Table 1.

[0022] Table 1 XRPD pattern analysis data of the crystal form of compound A of formula (I)

[0023] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned crystal form A has an endothermic peak starting point at 164.0±5°C.

[0024] In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form A is shown in Figure 2.

[0025] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form A shows a weight loss of 1.25% at 150.0±3°C.

[0026] In some embodiments of the present invention, the TGA spectrum of the above-mentioned crystal form A is shown in Figure 3.

[0027] The present invention also provides a compound of formula (II), whose structural formula is as follows:

[0028] The present invention also provides a crystalline form B of the compound of formula (II), which has an X-ray powder diffraction pattern (XRPD) having characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 14.59±0.20°, 19.11±0.20° and 21.75±0.20°;

[0029] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 19.11±0.20°, 21.75±0.20°, 24.94±0.20° and 25.73±0.20°.

[0030] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 16.62±0.20°, 19.11±0.20°, 21.75±0.20°, 22.23±0.20°, 24.94±0.20°, 25.73±0.20°, 26.22±0.20°, and 27.88±0.20°.

[0031] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 7.66±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 16.62±0.20°, 17.65±0.20°, 18.35±0.20°, 19.11±0.20°, 21.75±0.20°, 22.23±0.20°, 24.94±0.20°, 25.73±0.20°, 26.22±0.20°, and 27.88±0.20°.

[0032] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 7.66±0.20°, 8.11±0.20°, 11.34±0.20°, 12.21±0.20°, 13.89±0.20°, 14.59±0.20°, 14.84±0.20°, 15.31±0.20°, 16.20±0.20°, 16.62±0.20°, 17.44±0.20°, 17.65±0.20°, 18.35±0.20°, 19.11±0.20°, 19.84±0.20°, 20.43±0.20° , 21.75±0.20°, 22.23±0.20°, 22.46±0.20°, 23.50±0.20°, 24.36±0.20°, 24.94±0.20°, 25.73±0.20°, 26.22±0.20°, 27.43±0.20°, 27.88±0.20°, 28.72±0.20°, 29.14±0.20°, 30.11±0.20°, 30.64±0.20°, 32.04±0.20°, 32.87±0.20°, 33.46±0.20°, 35.89±0.20°, 38.78±0.20° and 39.28±0.20°.

[0033] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22±0.10°, 7.66±0.10°, 8.11±0.10°, 11.34±0.10°, 12.21±0.10°, 13.89±0.10°, 14.59±0.10°, 14.84±0.10°, 15.31±0.10°, 16.20±0.10°, 16.62±0.10° , 17.44±0.10°, 17.65±0.10°, 18.35±0.10°, 19.11±0.10°, 19.84±0.10°, 20.43±0.10°, 21.75±0.10°, 22.23±0.10°, 22.46±0.10°, 23.50±0.10°, 24.36±0.10°, 24.94±0.10°, 25.73±0.10°, 26.22±0.10°, 27.43±0.10°, 27.88±0.10°, 28.72±0.10°, 29.14±0.10°, 30.11±0.10°, 30.64±0.10°, 32.04±0.10°, 32.87±0.10°, 33.46±0.10°, 35.89±0.10°, 38.78±0.10° and 39.28±0.10°.

[0034] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22, 7.66, 8.11, 11.34, 12.21, 13.89, 14.59, 14.84, 15.31, 16.20, 16.62, 17.44, 17.65, 18.35, 19.11, 19 .84, 20.43, 21.75, 22.23, 22.46, 23.50, 24.36, 24.94, 25.73, 26.22, 27.43, 27.88, 28.72, 29.14, 30.11, 30.64, 32.04, 32.87, 33.46, 35.89, 38.78 and 39.28.

[0035] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned B crystal form has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, and / or 7.66±0.20°, and / or 11.34±0.20°, and / or 12.21±0.20°, and / or 13.89±0.20°, and / or 14.59±0.20°, and / or 14.84±0.20°. 0.20°, and / or 15.31±0.20°, and / or 16.20±0.20°, and / or 16.62±0.20°, and / or 17.44±0.20°, and / or 17.65±0.20°, and / or 18.35±0.20°, and / or 19.11±0.20°, and / or 19.84±0.20°, and / or 20.43±0.20°, and / or 21. 75±0.20°, and / or 22.23±0.20°, and / or 22.46±0.20°, and / or 23.50±0.20°, and / or 24.36±0.20°, and / or 24.94±0.20°, and / or 25.73±0.20°, and / or 26.22±0.20°, and / or 27.43±0.20°, and / or 27.88±0.20°, and / or 28.72±0.20°, and / or 29.14±0.20°, and / or 30.11±0.20°, and / or 30.64±0.20°, and / or 32.04±0.20°, and / or 32.87±0.20°, and / or 33.46±0.20°, and / or 35.89±0.20°, and / or 38.78±0.20°, and / or 39.28±0.20°.

[0036] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form B has characteristic diffraction peaks at the following 2θ angles: 6.22±0.10°, 8.11±0.10°, and / or 7.66±0.10°, and / or 11.34±0.10°, and / or 12.21±0.10°, and / or 13.89±0.10°, and / or 14.59±0.10°, and / or 14.84±0.10°. 0.10°, and / or 15.31±0.10°, and / or 16.20±0.10°, and / or 16.62±0.10°, and / or 17.44±0.10°, and / or 17.65±0.10°, and / or 18.35±0.10°, and / or 19.11±0.10°, and / or 19.84±0.10°, and / or 20.43±0.10°, and / or 21. 75±0.10°, and / or 22.23±0.10°, and / or 22.46±0.10°, and / or 23.50±0.10°, and / or 24.36±0.10°, and / or 24.94±0.10°, and / or 25.73±0.10°, and / or 26.22±0.10°, and / or 27.43±0.10°, and / or 27.88±0.10°, and / or 28.72±0.10°, and / or 29.14±0.10°, and / or 30.11±0.10°, and / or 30.64±0.10°, and / or 32.04±0.10°, and / or 32.87±0.10°, and / or 33.46±0.10°, and / or 35.89±0.10°, and / or 38.78±0.10°, and / or 39.28±0.10°.

[0037] In some embodiments of the present invention, the XRPD pattern of the above-mentioned Form B is basically as shown in Figure 4.

[0038] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form B is shown in Table 2.

[0039] Table 2 XRPD pattern analysis data of the crystal form of compound B of formula (II)

[0040] In some embodiments of the present invention, the above-mentioned Form B has a differential scanning calorimetry curve with an endothermic peak at 153.7±3°C.

[0041] In some embodiments of the present invention, the DSC spectrum of the above-mentioned Form B is shown in Figure 5.

[0042] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned Form B shows a weight loss of 2.88% at 150.0±3°C.

[0043] In some embodiments of the present invention, the TGA spectrum of the above-mentioned B crystal form is shown in Figure 6.

[0044] The present invention also provides a compound of formula (III), whose structural formula is as follows:

[0045] The present invention also provides a crystalline form C of the compound of formula (III), which has an X-ray powder diffraction pattern (XRPD) having characteristic diffraction peaks at the following 2θ angles: 5.46±0.20°, 13.19±0.20°, 15.22±0.20° and 20.59±0.20°;

[0046] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.46±0.20°, 13.19±0.20°, 15.22±0.20°, 15.80±0.20°, 16.25±0.20°, 18.12±0.20°, 20.59±0.20°, 21.63±0.20° and 26.50±0.20°.

[0047] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.46±0.20°, 13.19±0.20°, 15.22±0.20°, 15.80±0.20°, 16.25±0.20°, 18.12±0.20°, 20.59±0.20°, 21.63±0.20°, 24.11±0.20°, 26.50±0.20°, 27.75±0.20° and 28.15±0.20°.

[0048] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.46±0.20°, 9.30±0.20°, 13.19±0.20°, 15.22±0.20°, 15.80±0.20°, 16.25±0.20°, 18.12±0.20°, 20.59±0.20°, 21.63±0.20°, 24.11±0.20°, 24.91±0.20°, 25.87±0.20°, 26.50±0.20°, 27.75±0.20°, 28.15±0.20°, 30.73±0.20° and 31.74±0.20°.

[0049] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.46, 9.30, 13.19, 15.22, 15.80, 16.25, 18.12, 20.59, 21.63, 24.11, 24.91, 25.87, 26.50, 27.75, 28.15, 30.73 and 31.74.

[0050] In some embodiments of the present invention, the X-ray powder diffraction pattern of the above-mentioned crystal form C has characteristic diffraction peaks at the following 2θ angles: 5.46±0.20°, 15.22±0.20°, and / or 9.30±0.20°, and / or 13.19±0.20°, and / or 15.80±0.20°, and / or 16.25±0.20°, and / or 18.12±0.20°, and / or 20. 59±0.20°, and / or 21.63±0.20°, and / or 24.11±0.20°, and / or 24.91±0.20°, and / or 25.87±0.20°, and / or 26.50±0.20°, and / or 27.75±0.20°, and / or 28.15±0.20°, and / or 30.73±0.20°, and / or 31.74±0.20°.

[0051] In some embodiments of the present invention, the XRPD pattern of the above-mentioned Form C is basically as shown in Figure 7.

[0052] In some embodiments of the present invention, the XRPD pattern analysis data of the above-mentioned Form C are shown in Table 3.

[0053] Table 3 XRPD pattern analysis data of the crystal form of compound C of formula (III)

[0054] In some embodiments of the present invention, the differential scanning calorimetry curve of the above-mentioned crystal form C has endothermic peaks at 105.1±3°C and 216.0±3°C.

[0055] In some embodiments of the present invention, the DSC spectrum of the above-mentioned crystal form C is shown in Figure 8.

[0056] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned crystal form C shows a weight loss of 8.22% at 150.0±3°C.

[0057] In some embodiments of the present invention, the TGA spectrum of the above-mentioned C crystal form is shown in Figure 9.

[0058] The present invention also provides a pharmaceutical composition comprising a therapeutically effective dose of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as an active ingredient and a pharmaceutically acceptable excipient;

[0059] In some embodiments of the present invention, the dosage form of the pharmaceutical composition is selected from capsules, granules, injections, pills, syrups, powders, ointments, emulsions, solutions, suspensions or tinctures, and other variables are as defined in the present invention.

[0060] In some embodiments of the present invention, the above-mentioned excipients are selected from excipients, fillers, binders, humectants, disintegrants, solubilizers, absorption accelerators, adsorbents, diluents, solubilizers, emulsifiers, lubricants, wetting agents, glidants, suspending agents, flavoring agents, spices or mixtures thereof, and other variables are as defined in the present invention.

[0061] The present invention also provides a method for preparing the compound of formula (I), which is prepared as follows:

[0062] wherein R1 is selected from H, CH3, Boc, Cbz, Bn and PMB;

[0063] 1) When R1 is selected from H, the preparation method of the compound of formula (I) is selected from:

[0064] 2) When R1 is selected from CH3, the compound of formula (II-3) is the compound of formula (I);

[0065] 3) When R1 is selected from Boc, Cbz, Bn and PMB, the preparation method of the compound of formula (I) is selected from:

[0066] Wherein, the preparation method of the compound of formula (II-3) is selected from:

[0067] Method 1 includes:

[0068] Method 2 includes:

[0069] Method 3 includes:

[0070] in,

[0071] R1 is selected from H, CH3, Boc, Cbz, Bn and PMB;

[0072] R2 is selected from Bn, PMB, PNB and MOM;

[0073] R4 is selected from CH3, MOM, Bn, SEM and Ac;

[0074] R5 is selected from F, Cl, Br, I, OTf and OCH2CF3;

[0075] R 31Selected from B(OH)2; and BF3K;

[0076] R6 is selected from F, Cl, Br and I.

[0077] R7 is selected from Cl, Br, OTf, OCH2CF3 and OTs.

[0078] The present invention also provides a method for preparing the compound of formula (I),

[0079] It includes the following steps:

[0080] Step 1: react compound EE-3 to obtain compound EE-4.

[0081] Step 2: react compound EE-4 to obtain compound EE-5.

[0082] Step 3: Compound EE-5 and compound AA react to obtain compound H-1.

[0083] Step 4: reacting compound H-1 to obtain a compound of formula (I),

[0084] In some embodiments of the present invention, the method for preparing the compound of formula (I) comprises the following steps: Step 1: reacting compound EE-3 with reagents A and B to obtain compound EE-4,

[0085] in,

[0086] Reagent A is selected from chloromethyl ether, bromomethyl ether, iodomethyl ether and methoxymethyl methanesulfonate, preferably chloromethyl ether;

[0087] Reagent B is selected from potassium carbonate, cesium carbonate, sodium carbonate, lithium carbonate, sodium hydrogen, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide, potassium tert-butoxide, sodium tert-butoxide, sodium hydroxide, lithium hydroxide and potassium hydroxide, preferably potassium carbonate;

[0088] Solvent C is selected from DMF, DCM, THF, 2-MeTHF, EtOAc, i-PrOAc, NMP and dioxane, preferably DMF.

[0089] In some embodiments of the present invention, the method for preparing the compound of formula (I) comprises the following steps:

[0090] Step 2: Compound EE-4 is reacted with reagents D, E, F, and G to obtain compound EE-5.

[0091] in,

[0092] Reagent D is selected from bis-xanaphthene boronic acid ester and xanaphthene borane, preferably xanaphthene borane;

[0093] Reagent E is selected from Pd(dppf)Cl2, Pd(dppf)Cl2.CH2Cl2, bis(acetonitrile)palladium(II)dichloride, Pd(OAc)2 and Pd2(dba)3, preferably Pd(OAc)2;

[0094] Reagent F is selected from Sphos, Xphos, XantPhos and 2-(dicyclohexylphosphino)biphenyl, preferably 2-(dicyclohexylphosphino)biphenyl;

[0095] Reagent G is selected from potassium acetate, sodium acetate, TEA, DIPEA and potassium 2-ethylhexanoate, preferably TEA;

[0096] Solvent H is selected from dioxane, DMSO, DMF, NMP, DCM, THF, 2-MeTHF and water, preferably dioxane.

[0097] In some embodiments of the present invention, the method for preparing the compound of formula (I) comprises the following steps:

[0098] Step 3: Compound EE-5 and compound AA react with reagents I and J to obtain compound H-1.

[0099] in,

[0100] Solvent H is selected from dioxane, DMSO, DMF, NMP, DCM, THF, 2-MeTHF and water, preferably dioxane;

[0101] Reagent I is selected from Pd(OAc)2-BINAP, Pd2(dba)3-Xphos, Brettphos-Pd-G3, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd(dppf)Cl2CH2Cl2 and Pd(OAc)2-Sphos, preferably Pd(dppf)Cl2CH2Cl2;

[0102] Reagent J is selected from Cs2CO3, K2CO3, Na2CO3, K3PO4, KOAc, KF and TEA, preferably K2CO3.

[0103] In some embodiments of the present invention, the method for preparing the compound of formula (I) comprises the following steps:

[0104] Step 4: reacting compound H-1 with reagent K to obtain a compound of formula (I),

[0105] in,

[0106] Reagent K is selected from hydrochloric acid, hydrobromic acid, trifluoroacetic acid, sulfuric acid, formic acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid, preferably hydrochloric acid.

[0107] In some embodiments of the present invention, the method for preparing the compound of formula (I) comprises the following steps:

[0108] Step 1: react compound EE-3 with reagents A and B to obtain compound EE-4.

[0109] Step 2: Compound EE-4 is reacted with reagents D, E, F, and G to obtain compound EE-5.

[0110] Step 3: Compound EE-5 and compound AA react with reagents I and J to obtain compound H-1.

[0111] Step 4: reacting compound H-1 with reagent K to obtain a compound of formula (I),

[0112] in,

[0113] Reagent A is selected from chloromethyl ether, bromomethyl ether, iodomethyl ether and methoxymethyl methanesulfonate, preferably chloromethyl ether;

[0114] Reagent B is selected from potassium carbonate, cesium carbonate, sodium carbonate, lithium carbonate, sodium hydrogen, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide, potassium tert-butoxide, sodium tert-butoxide, sodium hydroxide, lithium hydroxide and potassium hydroxide, preferably potassium carbonate;

[0115] Solvent C is selected from DMF, DCM, THF, 2-MeTHF, EtOAc, i-PrOAc, NMP and dioxane, preferably DMF;

[0116] Reagent D is selected from bis-xanaphthene boronic acid ester and xanaphthene borane, preferably xanaphthene borane;

[0117] Reagent E is selected from Pd(dppf)Cl2, Pd(dppf)Cl2.CH2Cl2, bis(acetonitrile)palladium(II)dichloride, Pd(OAc)2 and Pd2(dba)3, preferably Pd(OAc)2;

[0118] Reagent F is selected from Sphos, Xphos, XantPhos and 2-(dicyclohexylphosphino)biphenyl, preferably 2-(dicyclohexylphosphino)biphenyl;

[0119] Reagent G is selected from potassium acetate, sodium acetate, TEA, DIPEA and potassium 2-ethylhexanoate, preferably TEA;

[0120] Solvent H is selected from dioxane, DMSO, DMF, NMP, DCM, THF, 2-MeTHF and water, preferably dioxane;

[0121] Reagent I is selected from Pd(OAc)2-BINAP, Pd2(dba)3-Xphos, Brettphos-Pd-G3, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd(dppf)Cl2CH2Cl2 and Pd(OAc)2-Sphos, preferably Pd(dppf)Cl2CH2Cl2;

[0122] Reagent J is selected from Cs2CO3, K2CO3, Na2CO3, K3PO4, KOAc, KF and TEA, preferably K2CO3;

[0123] Reagent K is selected from hydrochloric acid, hydrobromic acid, trifluoroacetic acid, sulfuric acid, formic acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid, preferably hydrochloric acid.

[0124] The present invention also provides a method for preparing an intermediate compound of formula (AA):

[0125] in,

[0126] R 51 selected from the group consisting of F, Cl, Br and I;

[0127] R 11 Selected from CH3, Boc, Cbz, Bn and PMB.

[0128] In some embodiments of the present invention, the preparation method of the intermediate compound of formula (AA) above, wherein R 51 Selected from Cl, R 11 is selected from Boc, and others are as defined in the present invention.

[0129] The present invention also provides a method for preparing an intermediate compound of formula (AA):

[0130] In some embodiments of the present invention, the preparation method of the compound of formula (I) is selected from:

[0131] Method 1 includes:

[0132] Method 2 includes:

[0133] Method 3 includes:

[0134] Method 4 includes:

[0135] Method 5 includes:

[0136] Method 6 includes:

[0137] Method seven includes:

[0138] Method 8 includes:

[0139] Method nine includes:

[0140] Method 10 includes:

[0141] Method 11 includes:

[0142] Method 12 includes:

[0143] Method 13 includes:

[0144] Method 14 includes:

[0145] The present invention also provides the use of the above-mentioned crystal form A of compound of formula (I), the above-mentioned compound of formula (II), the above-mentioned crystal form B of compound of formula (II), the above-mentioned pharmaceutical composition and the above-mentioned preparation method in the preparation of drugs for treating diseases related to NLRP3 inflammasome.

[0146] In some embodiments of the present invention, the aforementioned diseases associated with NLRP3 inflammasome are selected from NLRP3 inflammasome-associated neuroinflammatory diseases (such as brain inflammation, brain damage) and neurodegenerative diseases (such as Parkinson's disease, Alzheimer's disease, multiple sclerosis).

[0147] In some embodiments of the present invention, the aforementioned diseases associated with NLRP3 inflammasome are selected from NLRP3 inflammasome-associated neuroinflammatory diseases and neurodegenerative diseases.

[0148] In some embodiments of the present invention, the NLRP3 inflammasome-associated neuroinflammatory disease is selected from brain inflammation and brain injury.

[0149] In some embodiments of the present invention, the neurodegenerative disease is selected from Parkinson's disease, Alzheimer's disease and multiple sclerosis.

[0150] In some embodiments of the present invention, the neurodegenerative disease is selected from Parkinson's disease and Alzheimer's disease.

[0151] Technical Effects

[0152] The crystal form A of the compound of formula (I) and the crystal form B of the compound of formula (II) of the present invention are easy to obtain, have good physical and chemical stability, and have high industrial application value and economic value.

[0153] The process for synthesizing the compound of formula (I) and its intermediates provided by the present invention has the following beneficial effects: the raw materials are cheap and readily available, and the disadvantages of being difficult to separate and purify and not easy to industrialize are overcome.

[0154] The NLRP3 inhibitor provided by the present invention can effectively inhibit the activity of NLRP3 inflammasome and the activation of downstream caspase-1, thereby inhibiting the maturation and secretion of IL-1β, and has good pharmacokinetic properties. It can be used to treat diseases related to abnormal activation of NLRP3 inflammasome, such as Parkinson's disease and Alzheimer's disease.

[0155] Definition and Description

[0156] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be construed as ambiguous or unclear unless specifically defined, but rather should be understood in accordance with its ordinary meaning. When a trade name appears herein, it is intended to refer to the corresponding commercial product or its active ingredient.

[0157] It must be noted that, unless the context clearly indicates otherwise or is clearly contradicted by the context, the singular forms "a," "an," "the," and similar terms as used in the disclosure of the invention as used herein and in the appended claims (especially in the appended claims) are to be construed to include both the singular and the plural, thus, for example, reference to "the compound" includes reference to one or more compounds; and so on.

[0158] The term "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically pharmaceutically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically pharmaceutically acceptable carriers. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0159] The "compound of formula (I) or a pharmaceutically acceptable salt thereof" described in the present application may be a "pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof", further, may be a "pharmaceutical composition comprising the compound of formula (II)", or may be a "pharmaceutical composition comprising the compound of formula (III)". For example, the above-mentioned "administering the compound of formula (I) or a pharmaceutically acceptable salt thereof to an individual in need thereof" may be "administering a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof to an individual in need thereof", further, may be "administering a pharmaceutical composition comprising the compound of formula (II) to an individual in need thereof" or may be "administering a pharmaceutical composition comprising the compound of formula (III) to an individual in need thereof".

[0160] The term "therapeutically effective dose" means the amount of a compound that, when administered to a human for treating a disease, is sufficient to effect treatment for the disease.

[0161] The term "solvate" refers to a substance formed by a compound of the present invention or a salt thereof and a stoichiometric or non-stoichiometric amount of a solvent bound to the compound or salt thereof by non-covalent forces between the molecules. When the solvent is water, the solvate is a hydrate.

[0162] The term "treat" includes inhibiting, slowing, halting or reversing the progression or severity of an existing symptom or condition.

[0163] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0164] The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols.

[0165] Typical routes of administration of the compounds of the present invention, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0166] The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, sugar-coated pill making methods, grinding methods, emulsification methods, freeze-drying methods, etc.

[0167] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0168] The intermediate compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention.

[0169] The structures of the compounds of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compounds, the absolute configuration can be confirmed by conventional techniques in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data on the cultured single crystal using a Bruker D8 venture diffractometer, using CuKα radiation as the light source and scanning mode: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0170] The chemical reactions of the present invention are carried out in suitable solvents that are compatible with the chemical transformations of the present invention and the reagents and materials required. To obtain the compounds of the present invention, it may sometimes be necessary for those skilled in the art to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0171] An important consideration in planning any synthetic route in this field is the selection of an appropriate protecting group for a reactive functional group, such as the amino group in the present invention.

[0172] The present invention will be described in detail below through examples, which are not intended to limit the present invention in any way.

[0173] All solvents used in the present invention were commercially available and used without further purification.

[0174] The solvent used in the present invention is commercially available.

[0175] The present invention uses the following abbreviations: Boc represents tert-butyloxycarbonyl; Cbz represents benzyloxycarbonyl; Bn represents benzyl; PMB represents p-methoxybenzyl; DCM represents dichloromethane; DMF represents N,N-dimethylformamide; THF represents tetrahydrofuran; DMSO represents dimethyl sulfoxide; EtOH represents ethanol; MeOH represents methanol; ACN (MeCN) represents acetonitrile; EtOAc represents ethyl acetate; H2O represents water; i-PrOH represents isopropyl alcohol; Acetone represents acetone; IPAc represents isopropyl acetate; 2-MeTHF represents 2-methyltetrahydrofuran; 1,4-Dioxane represents 1,4-dioxane; CHCl3 represents chloroform; Toluene represents toluene; n-Heptane represents n-heptane; DMAc represents N,N-dimethylacetamide; NMP represents N-methylpyrrolidone; PNB represents p-nitrophenyl; MOM represents methoxymethyl; SEM represents 2-(trimethylsilyl)ethoxymethyl; Ac represents acetyl; OTf represents trifluoromethanesulfonate; OTs represents p-toluenesulfonate; i- PrOAc represents isopropyl acetate; Pd(dppf)Cl2.CH2Cl2 represents [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride in dichloromethane; Pd(dppf)Cl2 represents [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride; Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium; Sphos represents 2-dicyclohexylphosphino-2,6-dimethoxybiphenyl; Xphos represents 2-di-tert-butylphosphino-2′,4′,6′-triisopropylbiphenyl; TEA represents triethylamine; DIPEA represents diisopropylethylamine; BINAP represents 2,2-bis(diphenylphosphino)-11-binaphthyl; Pd(PPh3)2Cl2 represents bis(triphenylphosphine)palladium dichloride; Pd(PPh3)4 represents tetrakis(triphenylphosphine)palladium; Brettphos-Pd-G3 represents methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2,4,6-triisopropyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium; BID represents twice daily; QD represents once daily; po or PO represents oral administration.

[0176] Compounds are manually or Software naming, commercially available compounds use supplier catalog names.

[0177] Instrument test parameters and analysis methods of the present invention

[0178] The test parameters of the X-ray powder diffractometer (XRPD) method of the present invention are shown in Table 4.

[0179] Table 4 XRPD test parameters

[0180] The test parameters of the differential scanning calorimeter (DSC) method of the present invention are shown in Table 5.

[0181] Table 5 DSC test parameters

[0182] Thermogravimetric analysis (TGA) method of the present invention and test parameters are shown in Table 6.

[0183] Table 6 TGA test parameters

[0184] The test parameters of the dynamic vapor sorption analysis (DVS) method of the present invention are shown in Table 7.

[0185] Table 7 DVS test parameters BRIEF DESCRIPTION OF THE DRAWINGS

[0186] Figure 1: XRPD spectrum of the crystal form A of compound of formula (I) using Cu-Kα radiation;

[0187] Figure 2: DSC spectrum of the crystalline form A of compound of formula (I);

[0188] Figure 3: TGA spectrum of the crystalline form A of compound of formula (I);

[0189] Figure 4: XRPD spectrum of the crystal form B of compound of formula (II) using Cu-Kα radiation;

[0190] Figure 5: DSC spectrum of the crystalline form B of compound of formula (II);

[0191] Figure 6: TGA spectrum of the crystalline form B of compound of formula (II);

[0192] Figure 7: XRPD spectrum of the crystal form C of the compound of formula (III) using Cu-Kα radiation;

[0193] Figure 8: DSC spectrum of the crystalline form C of compound of formula (III);

[0194] Figure 9: TGA spectrum of the crystalline form C of compound of formula (III);

[0195] Figure 10: DVS spectrum of the crystalline form A of compound of formula (I);

[0196] Figure 11: DVS spectrum of the crystalline form B of compound of formula (II);

[0197] Figure 12: DVS spectrum of the crystalline form C of compound of formula (III);

[0198] Figure 13: PSD spectrum of the crystalline form A of compound of formula (I);

[0199] Figure 14: Single crystal X-ray diffraction (SC-XRD) ellipsoidal diagram of the three-dimensional structure of the compound of formula (I). DETAILED DESCRIPTION

[0200] In order to better understand the content of the present invention, further description is given below in conjunction with specific embodiments, but the specific implementation methods are not intended to limit the content of the present invention.

[0201] Example 1: Preparation of Intermediate AA

[0202] Synthesis route:

[0203] first step

[0204] Compound AA-1 (100 g, 691.76 mmol) and pyridine (109.98 g, 1.39 mol) were dissolved in anhydrous dichloromethane (1000 mL). Trifluoromethanesulfonic anhydride (255.68 g, 906.21 mmol) was added dropwise to the reaction mixture at 0°C. The reaction mixture was stirred at 25°C for 12 hours. Water (1000 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (500 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 100% to 90%) to obtain compound AA-2. 1 H NMR (400 MHz, CDCl3) δ 7.34 (s, 1H), 2.53 (s, 3H). MS-ESI calculated value [M+H] + 277, measured value 277.

[0205] Step 2

[0206] Compound AA-3 (117.29 g, 585.64 mmol) and compound AA-2 (90 g, 325.36 mmol) were dissolved in anhydrous DMF (90 mL). The reaction mixture was stirred at 90°C for 1 hour. Ethyl acetate (1000 mL) was added to the reaction mixture, filtered, and the filtrate was washed with sodium hydroxide (1% aqueous solution, 1000 mL) and water (500 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, 10 / 1) to obtain compound AA-4. MS-ESI calculated value [M+H] + 327, measured value 327.

[0207] Step 3

[0208] Dissolve compound AA-4 (53 g, 162.17 mmol) in ethyl acetate (500 mL). Add ethyl acetate hydrochloride (4 mol / L, 162.17 mL). Stir the reaction mixture at 25°C for 12 hours. Filter the mixture, wash the filter cake with ethyl acetate (300 mL), and vacuum dry the resulting mixture to obtain the hydrochloride salt of compound AA-5. 1 H NMR (400 MHz, CD3OD) δ7.53 (s, 1H), 4.20-4.18 (m, 1H), 3.60-3.57 (m, 1H), 3.34-3.31 (m, 1H), 3.13-3.10 (m, 2H), 2.48 (s, 3H), 2.31-2.21 (m, 1H), 2.15-2.05 (m, 1H), 2.01-1.95 (m, 1H), 1.79-1.77 (m, 1H). MS-ESI calculated value [M+H] + 227, measured value 227.

[0209] Step 4

[0210] Under nitrogen, the hydrochloride salt of compound AA-5 (85.0 g, 283.69 mmol) was dissolved in anhydrous methanol (850 mL). 37% aqueous formaldehyde solution (69.06 g, 851 mmol) was added to the reaction solution, and the reaction mixture was stirred at 20°C for 0.5 hours. Sodium acetate borohydride (123.97 g, 567 mmol) was then added portionwise at 20°C, and the reaction mixture was stirred at 20°C for 11.5 hours. Saturated aqueous ammonium chloride solution (300 mL) was added to the reaction solution, and the methanol was removed by concentration under reduced pressure. The remaining aqueous phase was adjusted to pH 8 with aqueous NaOH (3 mol / L). A large amount of solid precipitated and was filtered. The filter cake was washed with n-heptane (200 mL x 2) and dried under vacuum to obtain the crude product. The crude product was added to a mixed solvent of n-heptane: isopropyl acetate: NaOH aqueous solution (3 mol / L) (700 mL: 70 mL: 70 mL), suspended and stirred at 25 °C for 1 hour, filtered, and the filter cake was washed with (100 mL × 2) n-heptane and dried in vacuo to obtain compound AA. 1 H NMR (400 MHz, CD3OD) δ 6.78 (s, 1H), 4.10-3.97 (m, 1H), 2.99 (br d, J = 8.0 Hz, 1H), 2.66 (br d, J = 9.6 Hz, 1H), 2.27 (s, 3H), 2.26 (s, 3H), 2.16-2.07 (m, 1H), 2.05-1.91 (m, 2H), 1.82-1.74 (m, 1H), 1.72-1.60 (m, 1H), 1.44-1.20 (m, 1H). MS-ESI calcd. [M+H] +241, measured value 241.

[0211] Example 2: Preparation of intermediate EE-5

[0212] Synthesis route:

[0213] first step

[0214] A mixture of copper bromide (733.69 g, 3.28 mol) and ethyl acetate (1200 mL) was stirred at 80°C for 10 minutes. Compound EE-1 (125 g, 821.22 mmol) was dissolved in chloroform (1200 mL) and added to the reaction mixture. The mixture was heated and stirred at 80°C for 12 hours. After cooling, the reaction mixture was filtered through neutral alumina and celite. The filter cake was washed with dichloromethane (1000 mL), and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was then slurried with ethyl acetate (500 mL) to obtain compound EE-2. 1 H NMR (400 MHz, CDCl3) δ 7.48 (d, J = 5.2 Hz, 1H), 7.18 (d, J = 5.2 Hz, 1H), 3.20-3.15 (m, 4H). MS-ESI calculated value [M+H] + 311, measured value 311.

[0215] Step 2

[0216] Dissolve compound EE-2 (215 g, 693.54 mmol) in DMF (1000 mL) and add lithium carbonate (307.47 g, 4.16 mol) to the reaction mixture. Stir the reaction mixture at 100°C for 6 hours. Filter the reaction mixture, wash the filter cake with DMF (1000 mL), and concentrate the filtrate under reduced pressure to 600 mL. Add the residue to water (6000 mL) and adjust the pH to 1 with dilute hydrochloric acid (1 mol / L). A large amount of solid will precipitate. Stir at room temperature for 30 minutes, filter, and dry to obtain compound EE-3. 1 H NMR (400MHz, CDCl3) δ7.51 (d, J = 5.6 Hz, 1H), 7.45-7.37 (m, 2H), 7.34 (d, J = 8.8 Hz, 1H), 5.87 (s, 1H).

[0217] Step 3

[0218] Compound EE-3 (200 g, 873.01 mmol) and potassium carbonate (301.65 g, 2.18 mol) were dissolved in DMF (1000 mL). The reaction solution was cooled to 0°C, and chloromethyl ether (153.070 g, 1.90 mol) was slowly added dropwise to the reaction solution. The reaction solution was allowed to react at 25°C for 12 hours. The reaction solution was added to ice water (5000 mL) with stirring, and the pH was adjusted to 10 with saturated potassium carbonate. The solution was extracted with ethyl acetate (1000 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The concentrate was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 200 / 1 to 100 / 1) to obtain compound EE-4. 1 H NMR (400MHz, CD3OD) δ7.61-7.56(m,2H),7.53-7.46(m,2H),5.24(s,2H),3.66(s,3H).

[0219] Step 4

[0220] Compound EE-4 (100 g, 366.10 mmol, 1 eq), compound 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (70.28 g, 549.16 mmol), triethylamine (111.14 g, 1.10 mol), palladium acetate (6.58 g, 29.29 mmol), and 2-(dicyclohexylphosphino)biphenyl (20.53 g, 58.58 mmol, 0.16 eq) were dissolved in dioxane (1000 mL). The mixture was stirred at 100°C under nitrogen for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent, and the residue was dissolved in ethyl acetate (500 mL), filtered through a silica gel pad, and the filtrate was concentrated under reduced pressure. The concentrate was separated by silica gel column chromatography (petroleum ether / ethyl acetate = 200 / 1 to 100 / 1) to obtain compound EE-5. 1 H NMR (400MHz, CD3OD) δ7.69-7.65(m,2H),7.57-7.52(m,2H),5.24(s,2H),3.59(s,3H),1.39(s,12H).

[0221] Example 3: Preparation of compound of formula (I)

[0222] Synthesis route:

[0223] first step

[0224] Under nitrogen protection, compound AA (64.0 g, 265.86 mmol) and compound EE-5 (102.16 g, 319 mmol) were dissolved in dioxane (378 mL), and a solution of potassium carbonate (73.49 g, 531.71 mmol) in water (252 mL) was slowly added. 1,1-bis(diphenylphosphino)ferrocenepalladium dichloride (4.34 g, 5.32 mmol) was added to the reaction solution. The reaction solution was stirred at 85 ° C for 3 hours. The reaction solution was filtered with diatomaceous earth, and the filter cake was washed with 2-methyltetrahydrofuran (200 mL×3). The filtrate was concentrated under reduced pressure to remove dioxane and 2-methyltetrahydrofuran. The remaining aqueous phase was adjusted to pH 4 with aqueous hydrochloric acid solution (1 mol / L), extracted with 2-methyltetrahydrofuran (1000 mL×2), the aqueous phase was adjusted to pH 10 with aqueous sodium hydroxide solution (1 mol / L), extracted with 2-methyltetrahydrofuran (1000 mL×1, 700 mL×2), the organic phases were combined and concentrated, the organic phase was dissolved with 1400 ml of dimethyltetrahydrofuran, modified silica gel for palladium removal (LI-511, 50 g) was added, stirred at 45°C for 6 hours, filtered, and modified silica gel for palladium removal (LI-511, 50 g) was added to the filtrate, stirred at 50°C for 12 hours, filtered, modified silica gel for palladium removal (LI-511, 50 g) was added to the filtrate, stirred at 50°C for 6 hours, filtered, modified silica gel for palladium removal (iMoLboX-LMat-NH01x, 50 g) was added to the filtrate, stirred at 50°C for 12 hours, filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product. The crude product was added to a mixed solvent of n-heptane: isopropyl acetate (700 mL: 70 mL), suspended and stirred at 25°C for 30 minutes. After the suspension was filtered, the filter cake was washed with n-heptane (100 mL × 3), and the filter cake was vacuum dried to obtain compound I-1. 1 H NMR (400 MHz, CD3OD) δ 7.79 (d, J = 8.4 Hz, 1H), 7.64 (d, J = 5.6 Hz, 1H), 7.54 (d, J = 5.6 Hz, 1H), 7.27 (d, J = 8.4 Hz, 1H), 6.80 (s, 1H), 4.94 (s, 2H), 4.24-4.03 (m, 1H), 3.15 (s, 3H), 3.09 (br d, J = 5.2 Hz, 1H), 2.71 (br d, J = 6.0 Hz, 1H), 2.30 (s, 3H), 2.19-2.01 (m, 6H), 1.87-1.64 (m, 2H), 1.39-1.36 (m, 1H). MS-ESI calcd. [M+H] + 399, measured value 399.

[0225] Step 2

[0226] Compound I-1 (73.0 g, 183.18 mmol) was added portionwise to hydrochloric acid (2 mol / L, 730.0 mL) at 10°C. The reaction mixture was stirred at 25°C for 6 hours. The reaction mixture was diluted with 365 mL of water and 292 mL of dichloromethane. Ammonia was added at 10°C to adjust the pH to 8 to 9. The mixture was then separated and the aqueous phase was extracted with dichloromethane (146 mL x 2). The organic phase was dried, filtered, and concentrated under reduced pressure to obtain an amorphous form of the compound of formula (I). 1 H NMR (400 MHz, CD3OD) δ 7.58 (d, J = 5.2 Hz, 1H), 7.48-7.46 (m, 2H), 7.23 (d, J = 8.4 Hz, 1H), 6.79 (s, 1H), 4.19-3.99 (m, 1H), 3.04 (br d, J = 6.0 Hz, 1H), 2.68 (br d, J = 7.2 Hz, 1H), 2.30 (s, 3H), 2.22-2.12 (m, 4H), 2.10-1.95 (m, 2H), 1.91-1.77 (m, 1H), 1.76-1.62 (m, 1H), 1.39-1.36 (m, 1H). MS-ESI calcd. [M+H] + 355, measured value 355.

[0227] Example 4: Preparation of Crystalline Form A of Compound (I)

[0228] The amorphous form of the compound of formula (I) (150 mg, 423.17 μmol) was added to acetonitrile (1.5 mL), suspended and stirred at 50°C for 24 hours, cooled to room temperature, and filtered. The filter cake was washed with acetonitrile (1 mL) and dried under vacuum to obtain Form A of the compound of formula (I). The XRPD, DSC, TGA, DVS, and PSD analysis results are shown in Figures 1, 2, 3, 10, and 13, respectively.

[0229] The acetonitrile solvent was replaced with water, isopropanol, acetonitrile, acetone, ethyl acetate, isopropyl acetate, 2-methyltetrahydrofuran, tetrahydrofuran, methyl isobutyl ketone, acetonitrile / water (1:1), acetonitrile / water (1:4), acetone / water (1:1), and acetone / water (1:4), all yielding Form A of the compound of formula (I). The experimental results are shown in Table 8.

[0230] Table 8 50℃ suspension stirring test

[0231] Example 5: Preparation of Form B of the compound of formula (II)

[0232] The amorphous form (1.0 g, 2.82 mmol) of the compound of formula (I) was added to acetone (10 mL) and stirred to dissolve. D-malic acid (378.28 mg, 2.82 mmol) in acetone was added to the above solution. Acetone (20 mL) was continued to be added to the reaction solution. The reaction solution was suspended and stirred at 50°C for 24 hours, cooled to room temperature, filtered, and the filter cake was vacuum dried to obtain Form B of the compound of formula (II). 1 H NMR(400MHz,CD3OD)δ7.62(d,J=5.6Hz,1H),7.55-7.51(m,2H),7.24(d,J=8.0Hz, 1H),6.92(s,1H),4.47-4.39(m,1H),4.34(dd,J=7.6,5.2Hz,1H),3.74-3.60(m,1 H), 3.22-3.19 (m, 1H), 3.08-2.87 (m, 2H), 2.85-2.80 (m, 1H), 2.79 (s, 3H), 2.61-2.54 (m, 1H), 2.21 (s, 3H), 2.16-2.06 (m, 2H), 2.00-1.88 (m, 1H), 1.75-1.69 (m, 1H). The results of XRPD, DSC, TGA, and DVS are shown in Figures 4, 5, 6, and 11, respectively.

[0233] Example 6: Preparation of Form C of the compound of formula (III)

[0234] The amorphous form of the compound of formula (I) was prepared using high-performance liquid chromatography (HPLC column: Phenomenex C18, 150×40mm×5μm; mobile phase: [water (hydrochloric acid)-acetonitrile]; B%: 1%-30%, 10 minutes) to obtain the amorphous form of the compound of formula (III). Acetone (0.5mL) was added to the amorphous form of the compound of formula (III) (50.2mg), and the resulting suspension was stirred at room temperature for 3 days to obtain Form C of the compound of formula (III). The results of XRPD, DSC, TGA, and DVS are shown in Figures 7, 8, 9, and 12, respectively.

[0235] Example 7: Solid Preliminary Stability Test of Compound A of Formula (I) and Compound B of Formula (II)

[0236] According to the "Guidelines for Stability Testing of APIs and Preparations" (Chinese Pharmacopoeia 2020 Edition Part IV General Rules 9001), the stability of the crystalline form of compound A of formula (I) and the crystalline form of compound B of formula (II) were investigated under high temperature (60°C, open), high humidity (25°C / relative humidity 92.5%, open), strong light (5000lx and ultraviolet intensity 90μw / cm 2, open) and pre-stability under high temperature and high humidity conditions (40℃ / relative humidity 75% and 60℃ / relative humidity 75% or 30℃ / relative humidity 65%, open).

[0237] Weigh about 30 mg of each of the crystal form A of compound (I) and the crystal form B of compound (II), place them in a dry and clean glass bottle, spread them into a thin layer, and use them as test samples. Place them under influencing factors and accelerated conditions, and the samples are completely exposed. Samples are taken for analysis after 5 days and 10 days under high temperature and high humidity conditions, and after 1 month, 2 months, and 3 months under accelerated conditions. Samples placed under light conditions are completely exposed at room temperature. XRPD is performed on samples placed under different conditions, and the test results are compared with the initial test results on day 0. The crystal form results are shown in Tables 9 and 10 below. The results show that the crystal form A of compound (I) and the crystal form B of compound (II) are stable crystal forms.

[0238] Table 9 Preliminary stability results of the crystal form of compound A of formula (I)

[0239] Table 10 Preliminary stability results of the crystal form of compound B of formula (II)

[0240] Example 8 Hygroscopicity Study of Compound A (Formula (I), Compound B (Formula (II)) and Compound C (Formula (III))

[0241] Experimental Materials:

[0242] Dynamic Vapor Adsorption Analyzer

[0243] Experimental methods:

[0244] About 10 mg of the compound was weighed and placed in a DVS sample pan for testing.

[0245] The classification of moisture absorption evaluation is shown in Table 11:

[0246] Table 11 Moisture absorption evaluation classification table Note: ΔW% indicates the weight gain of the test sample at 25±1℃ and 80±2%RH.

[0247] Experimental results:

[0248] The DVS spectrum of Form A of Compound of Formula (I) is shown in Figure 10. The DVS results show that the sample absorbed moisture at 25°C / 80% RH, resulting in a weight gain of 0.1665%, indicating that the sample has little or no hygroscopicity. After completing the DVS test (0-95-0% RH), the sample was exposed to air and analyzed by XRPD. The results showed that the crystalline form remained unchanged before and after the DVS test.

[0249] The DVS spectrum of the crystalline form B of compound (II) is shown in Figure 11. The DVS results show that the sample has a moisture absorption weight increase of 7.15% at 25°C / 80% RH compared to 25°C / 0% RH, indicating that the sample is hygroscopic.

[0250] The DVS spectrum of the crystalline form C of the compound of formula (III) is shown in Figure 12. The DVS results show that the sample has a moisture absorption weight increase of 21.09% at 25°C / 80% RH compared to 25°C / 0% RH, indicating that the sample is highly hygroscopic.

[0251] Experimental conclusion:

[0252] The crystal form A of the compound of formula (I) has no or almost no hygroscopicity at 25±1°C and 80±2% RH, and the crystal form remains unchanged before and after; the crystal form B of the compound of formula (II) is hygroscopic at 25±1°C and 80±2% RH; and the crystal form C of the compound of formula (III) is extremely hygroscopic at 25±1°C and 80±2% RH.

[0253] Example 9 Particle size distribution and morphology research test

[0254] The particle size distribution test parameter settings are shown in Table 12, and the experimental results are shown in Table 13.

[0255] Table 12 Test parameters of particle size distribution *: A flow rate of 60% is 60% of 65 mL / s.

[0256] Table 13 PSD results of the crystal form of compound A of formula (I)

[0257] MV: mean particle size calculated by volume;

[0258] SD: stands for standard deviation;

[0259] D10: represents the particle size corresponding to 10% of the particle size distribution (volume distribution);

[0260] D50: indicates the particle size corresponding to 50% of the particle size distribution (volume distribution), also known as the median diameter;

[0261] D90: indicates the particle size corresponding to 90% of the particle size distribution (volume distribution);

[0262] Experimental conclusion:

[0263] The PSD results of the crystalline form A of compound (I) are shown in FIG13 . The volume average particle size of the crystalline form A of compound (I) is 12.05 μm. The particle size distribution is relatively narrow, almost showing a normal distribution, and the particle size distribution is uniform.

[0264] Example 10: Single crystal X-ray diffraction analysis of the compound of formula (I)

[0265] 1. Manufacturer and instrument parameters of the single crystal X-ray diffractometer

[0266] Manufacturer: Bruker;

[0267] Instrument model: D8 VENTURE

[0268] X-ray light source: high-intensity micro-focus rotating anode light source, Cu target;

[0269] Power 2.5KW;

[0270] Tube voltage 50kV;

[0271] Tube current 50mA;

[0272] Goniometer: Four-axis (Kappa, ω, 2θ, ) goniometer;

[0273] Detector: Large-area photon II detector, with an effective area of ​​14cm×10cm. The distance from the detector to the sample is automatically adjustable by a motor.

[0274] 2. Crystal Cultivation

[0275] A sample of the compound of formula (I) (15 mg) was weighed and dissolved in toluene (0.6 mL) at room temperature. Petroleum ether (0.05 mL) was then added and filtered through a filter. The sample solution was placed in a 2 mL sample bottle and allowed to stand at room temperature for 2 weeks to culture to obtain single crystals.

[0276] 3. Conclusion

[0277] The single crystal SC-XRD ellipsoidal structure diagram of the compound of formula (I) is shown in Figure 14. The results show that C15 and C34 in the figure are in R configuration.

[0278] Biological test data:

[0279] Experimental Example 1: LPS-stimulated IL-1β release in rat primary microglia

[0280] 1. Experimental purpose:

[0281] In this study, primary rat microglia were used to study the inhibitory activity of NLRP3 inhibitors on IL-1β secretion in primary rat microglia.

[0282] 2. Experimental reagents: See Table 14 for a detailed list of experimental reagents.

[0283] Table 14 Experimental reagent list

[0284] 3. Experimental Operation

[0285] From newborn Sprague-Dawley rats, the cerebral cortex was harvested, digested, and isolated to obtain mixed glial cells, which were then inoculated into culture flasks for culture. The medium was changed every 3-4 days. After approximately 10 days of culture, the cells were shaken at 37°C, centrifuged, and harvested for microglial cells. The cells were then inoculated into 96-well cell culture plates and cultured overnight. The cells were then switched to serum-free medium and treated with 50 ng / mL LPS for 3 hours. Various concentrations of compounds were then added for 0.5 hours, followed by 0.3 μg / mL Nigericin for 1 hour. The supernatant was collected and stored at -80°C, or IL-1β release was assayed directly by ELISA according to the kit instructions. Data processing: The 10 μM positive compound group was used as the low value (L), and the DMSO group was used as the high value (H). The inhibition rate of the compound was calculated by the following formula (Inhibition% = (Ave_H-Sample) / (Ave_H-Ave_L) * 100), and the nonlinear regression equation Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)) calculates the IC of the compound 50 .

[0286] IV. Experimental results are shown in Table 15

[0287] Table 15 IC of NLRP3 inhibitors detected in primary rat microglia 50 Experimental test results

[0288] Conclusion: The compounds of the present invention have significant inhibitory activity on the maturation and secretion of IL-1β in rat primary microglial cells.

[0289] Experimental Example 2: Study on the efficacy of LPS-induced central nervous system inflammation model in rats

[0290] 1. Experimental purpose:

[0291] A central nervous system inflammation model was established by intracerebroventricular injection of LPS in rats, and the cerebrospinal fluid IL-1β level was evaluated by ELISA to assess the in vivo efficacy of the test compounds.

[0292] 2. Experimental Methods

[0293] SD male rats, 10-14 weeks old, were anesthetized with isoflurane inhalation. The animals' heads were maintained motionless and the brain surface was flattened. The rats' heads were shaved, and a skin incision was made along the sagittal suture to expose the bregma. A glass electrode was positioned at the bregma, and the axes of the coordinate display were reset to zero, and the electrodes were positioned according to the coordinates. After slowly inserting the needle into the lateral ventricle, 12.5 μg of LPS was injected, followed by 5 mg / kg of the compound of formula (I) 2 hours later and ATP 3 hours later. Cerebrospinal fluid was collected 3.5 hours later, and IL-1β inhibition levels were evaluated by ELISA.

[0294] 3. Experimental results are shown in Table 16

[0295] Table 16 Results of the IL-1β inhibitory activity experiment in rat cerebrospinal fluid Note: Mean ± SEM, n = 3; compared with the vehicle group: *p < 0.05, ***p < 0.001.

[0296] The results showed that a single dose of 5 mg / kg of the compound of formula (I) had inhibitory activity against central IL-1β, demonstrating superior in vivo efficacy.

[0297] Experimental Example 3: Pharmacological Study on 6-Hydroxydopamine-Induced Parkinson's Disease Model in Rats

[0298] 1. Experimental purpose:

[0299] SD rats were selected and a Parkinson's disease (PD) model was established by stereotaxic injection of 6-hydroxydopamine (6-OHDA). Five days after treatment with the compound, the striatum was collected and the IL-1β inhibition level was evaluated by ELISA.

[0300] II. Detailed list of experimental subjects, grouping and medication regimen is shown in Table 17

[0301] Table 17 Detailed list of experimental subjects, groups and dosing regimens

[0302] 3. Experimental methods:

[0303] Eight-week-old male Sprague-Dawley rats were anesthetized with isoflurane inhalation to prevent head movement and to flatten the brain surface. The rats' heads were shaved, and a skin incision was made along the sagittal suture to expose the bregma. A glass electrode was positioned at the bregma, and the coordinate display was reset to zero. The substantia nigra (SN) and striatum (Str) were located according to the coordinates. After slowly inserting the needle into the SN and Str, 6-OHDA was injected at a rate of 0.4 μL / min. After administration, the needle was held for another 10 minutes before slowly withdrawing the needle. The test compound (compound of Formula (I), 5 and 10 mg / kg) and the vehicle control (solvent stock solution) were administered starting the day before modeling and administered daily by gavage (BID or QD) until the end of the experimental period. Six days after drug treatment, samples were collected 2 hours after drug administration from half of the striatum on the modeling side. IL-1β levels in the rat striatum were measured using an ELISA kit.

[0304] IV. Experimental results are shown in Table 18

[0305] Table 18 Results of rat striatal IL-1β inhibitory activity experiment Note: Mean ± SEM, n = 4; compared with the 6-OHDA model group: ***p < 0.001.

[0306] The results showed that the test compound had a significant inhibitory effect on the level of IL-1β in the striatum of the 6-hydroxydopamine-induced Parkinson's disease rat model.

[0307] Experimental Example 4: Behavioral and Histological Detection of 6-Hydroxydopamine-Induced Parkinson's Disease Model in Rats

[0308] 1. Experimental purpose:

[0309] A Parkinson's disease (PD) model was established in SD rats via stereotaxic injection of 6-hydroxydopamine (6-OHDA). After 21 days of compound treatment, behavioral assessments (balance beam test, rotarod test) and histological tests (immunofluorescence staining for tyrosine hydroxylase (TH) and microglial marker (Iba1)) were performed to evaluate the efficacy of the test compounds.

[0310] 2. Experimental subjects and groups:

[0311] SD male rats (180-220 g). The specific dosing regimen is shown in Table 19.

[0312] Table 19 Detailed list of drug administration regimens for experimental subjects

[0313] 3. Experimental methods:

[0314] Sprague-Dawley rats were anesthetized with isoflurane inhalation, ensuring that the animal's head did not move and that the brain surface was flat. The rat's head was shaved, and a skin incision was made along the sagittal suture to expose the bregma. A glass electrode was positioned at the bregma, and the coordinate display was reset to zero. The substantia nigra (SN) and striatum (Str) brain regions were located according to the coordinates. After slowly advancing the needle to the SN and striatum, 6-OHDA was injected at a rate of 0.4 μL / min after a 10-minute wait. After administration, the needle was allowed to rest for another 10 minutes before slowly withdrawing.

[0315] During the balance beam test training phase, the animal was placed on the balance beam for 10 minutes daily, crossing the beam twice during each training session for a total of two days. During the formal test, the animal was placed on the balance beam for 30-60 minutes of acclimatization, and the number of foot slips during the two successful crosses was recorded. Evaluation criteria were the number of foot slips (foot leaving the top surface of the beam) during two successful crosses.

[0316] Rotarod test (muscle strength test): Animals were placed in the testing room for 30-60 minutes to acclimate to the environment before testing. Acclimation training involved placing each animal on a rotarod fatigue apparatus for 5 minutes. For the final test, the apparatus was set to a speed of 20 rpm / min and a test duration of 5 minutes. Rats were placed on the rotarod in batches for testing. Results analysis: The time each animal spent on the rod was recorded.

[0317] The day after the behavioral test, samples were collected for immunofluorescence staining of tyrosine hydroxylase (TH) and microglial marker (Iba1).

[0318] After data aggregation and statistics, SPSS statistical software was used for analysis (one-way analysis of variance), and graphs were drawn using Graph Pad software based on the SPSS analysis results. Immunohistochemistry images were drawn using Adobe Photoshop and Adobe Illustrator software.

[0319] 4. The experimental results are shown in Table 20, Table 21, Table 22, Table 23 and Table 24.

[0320] Table 20 Statistical analysis results of the number of foot slips in the balance beam test of rats 21 days after stereotaxic injection of 6-OHDA Note: Mean ± SEM, n = 12-15; *: Compared with the 6-OHDA model group, *p < 0.05, ***p < 0.001

[0321] The results showed that in the balance beam test, the rats in the compound group (I) slipped less than those in the 6-OHDA model group, and the difference was significant (p<0.05).

[0322] Table 21 Statistical analysis results of the time spent on the rod in the rotarod test of rats 21 days after stereotaxic injection of 6-OHDA Note: Mean ± SEM, n = 12-15; *: Compared with the 6-OHDA model group, *p < 0.05, ***p < 0.001

[0323] The results showed that compared with the 6-OHDA model group, the rats in the formula (I) compound group spent longer time on the rod than the rats in the 6-OHDA model group, and the difference between the two groups was significant (p<0.05).

[0324] Table 22 Effect of compound administration for 21 days on the number of signals in the SN brain region of rats in TH staining Note: Mean ± SEM, n = 12-15; *: Compared with the 6-OHDA model group, ***p < 0.001

[0325] The results showed that tyrosine hydroxylase (TH) staining was performed 21 days after the rats were stereotactically microinjected with 6-OHDA (20 μg / 8 μL) in the striatum and substantia nigra. In the SN brain region, the number of TH-positive cells in the blank group, the compound of formula (I) group and the 6-OHDA modeling group was significantly different (p<0.001).

[0326] Table 23 Effects of compound administration for 21 days on the number of signals in the SN brain region of rats in Iba-1 staining Note: Mean ± SEM, n = 12-15; *: Compared with the 6-OHDA model group, ***p < 0.001

[0327] Table 24 Effects of compound administration for 21 days on the number of Str signals in rat brain region in Iba-1 staining Note: Mean ± SEM, n = 12-15; *: Compared with the 6-OHDA model group, ***p < 0.001

[0328] The results showed that in the Str and SN brain regions, the number of Iba1-positive cells in the 6-OHDA modeling group was significantly increased compared with the blank group, and the difference was extremely significant (p<0.001), indicating that a significant inflammatory response was successfully induced in the SN and Str brain regions after PD modeling; after drug treatment, the difference between the formula (I) compound group and the 6-OHDA modeling group was extremely significant (p<0.001).

[0329] Experimental Example 5: Alzheimer's mouse model induced by humanized oligomeric Aβ1-42 (oAβ1-42) and efficacy experiment of the test product

[0330] 1. Experimental purpose:

[0331] C57BL / 6 mice were selected and an Alzheimer's Disease (AD) mouse model was established by intracerebroventricular injection of humanized oligomeric peptide Aβ1-42 (oAβ1-42), and the in vivo efficacy of the test compound was evaluated.

[0332] 2. Experimental subjects, grouping and medication regimen are shown in Table 25.

[0333] Table 25: Experimental animal groups, model induction and drug treatment

[0334] III. Experimental Methods: Intraventricular Injection in Mice: After light anesthesia with chloral hydrate, the heads of all animals in each group were fixed. The skin on the skull was wiped with 3% H₂O₂. The lateral ventricle was located using a mouse brain atlas. Using a stereotaxic technique (M / L = -0.9 mm, A / P = -0.2 mm, D / V = -2.35 mm), a sterile microsyringe was inserted perpendicularly into the skull. Groups 2 and 3 were slowly injected with oAβ1-42 solution (100 μM / 5 μL / mouse) at a rate of 0.8 μL / min. The needle was retained for 5 minutes after injection and then slowly removed for approximately 5 minutes. Group 1, the control group, received an equal volume of blank vehicle. The day of animal modeling was designated as day 0.

[0335] Treatment: Animals were administered drug or vehicle starting on day 1. The vehicle group and the oAβ1-42 model group received the vehicle orally, while the third group received the test compound solution orally twice daily, 8 hours apart. Behavioral experiments were conducted on days 8 and 10, and the second dose was administered after the behavioral experiment. Tissue samples were collected from mice 2 hours after the first dose on day 13.

[0336] Y-maze test: Mice were placed at either end of a Y-maze arm and allowed to freely explore for 8 minutes. The animal's movements and behavioral changes were recorded using video. The following metrics were recorded: ① Total number of entries: the number of times the animal entered an arm (one entry was defined as entering an arm with all four legs); ② Alternation: the number of consecutive entries into all three arms of the Y-maze; and ③ Maximum number of alternations: total number of entries minus 2. The final metric, spontaneous alternation score, was calculated as: total number of alternations / maximum number of alternations / 100%. Mouse behavior was analyzed using the EthosVision-XT8.0 video analysis system.

[0337] Drug administration continued after the behavioral test. On the thirteenth day, samples were collected from the hippocampus and cortex to detect the levels of inflammatory factors by ELISA; immunofluorescence staining was performed for phosphorylated Tau protein (AT8) and microglial marker (Iba-1) in the hippocampus and cortex.

[0338] 4. Data Analysis Methods

[0339] All immunofluorescence images were analyzed using Image J software for positive cell count analysis. All experimental results are presented as mean ± SEM, and data were analyzed using SPSS 20.0 software. Intergroup comparisons were performed using the independent-sample t-test to determine overall significant differences. Homogeneity of variance was assessed using the Levene's test; homogeneity was considered when p > 0.05. Significant differences between groups were determined using the t-test using the mean equation; significant differences were observed when p < 0.05.

[0340] 5. The experimental results are shown in Table 26, Table 27, Table 28, Table 29 and Table 30.

[0341] Table 26 Statistical analysis of spontaneous alternation behavior scores in the Y maze of mice 10 days after intracerebroventricular injection of oAβ1-42 Note: Data are expressed as Mean ± SEM; n = 14; *: compared with the vehicle group, **p < 0.01; # : Compared with the oAβ1-42 model group, # p< 0.05.

[0342] The results showed that the spontaneous alternation behavior score of the Y maze test compound group was significantly improved compared with the oAβ1-42 model group, suggesting that compound (I) has a significant improvement effect on the spatial learning and memory impairment of mice caused by oAβ.

[0343] Table 27 Results of inflammatory factor levels in the hippocampus 13 days after intraventricular injection of oAβ1-42 Note: Data are expressed as Mean ± SEM; *: Compared with the vehicle group, ***p < 0.001, ****p < 0.0001; # : Compared with the oAβ1-42 model group, ## p<0.01, ### p<0.001.

[0344] The results showed that the levels of IL-6 and IL-1β in the hippocampus of the compound group were significantly reduced compared with the oAβ1-42 model group, suggesting that the compound of formula (I) has a significant improvement effect on inflammatory damage in the hippocampus of mice caused by oAβ.

[0345] Table 28 Results of inflammatory factor levels in the cortical brain 13 days after intraventricular injection of oAβ1-42 Note: Data are expressed as Mean ± SEM; *: compared with the vehicle group, **p < 0.01, ****p < 0.0001; # : Compared with the oAβ1-42 model group, ### p<0.001.

[0346] The results showed that compared with the oAβ1-42 model group, the IL-6 level in the cortical area of ​​the compound group was significantly reduced, and the IL-1β level was slightly reduced, indicating that the compound of formula (I) has a significant improvement effect on the inflammatory damage in the mouse hippocampus caused by oAβ.

[0347] Table 29 Analysis of Tau protein phosphorylation levels and microglial activation in the hippocampus 13 days after intraventricular injection of oAβ1-42 Note: Data are expressed as Mean ± SEM; *: compared with the vehicle group, ****p < 0.0001; # : Compared with the oAβ1-42 model group, #### p < 0.0001.

[0348] The results showed that compared with the oAβ1-42 model group, the AT8 expression level and the proportion of activated Iba-1 microglia in the hippocampus of the compound group were significantly decreased, suggesting that the compound of formula (I) has a significant inhibitory effect on the hyperphosphorylation of hippocampal Tau protein and excessive activation of microglia caused by oAβ.

[0349] Table 30 Analysis of Tau protein phosphorylation levels and microglial activation in the cortical brain 13 days after intraventricular injection of oAβ1-42 Note: Data are expressed as Mean ± SEM; *: compared with the vehicle group, ****p < 0.0001; # : Compared with the oAβ1-42 model group, #### p < 0.0001.

[0350] The results showed that compared with the oAβ1-42 model group, the AT8 expression level and the proportion of activated Iba-1 microglia in the cortical brain region of the compound group were significantly decreased, suggesting that the compound of formula (I) has a significant inhibitory effect on the hyperphosphorylation of Tau protein and excessive activation of microglia in the cortical region caused by oAβ.

Claims

1. Form A of the compound of formula (I), having an X-ray powder diffraction pattern (XRPD) with characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 16.53±0.20°, and 18.12±0.20°; 2. The crystal form A according to claim 1, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 13.48±0.20°, 16.53±0.20°, 18.12±0.20°, 21.44±0.20° and 24.06±0.20°.

3. The crystal form A according to claim 2, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 8.41±0.20°, 12.72±0.20°, 13.48±0.20°, 16.53±0.20°, 18.12±0.20°, 21.44±0.20°, 24.06±0.20° and 25.55±0.20°.

4. The crystal form A according to claim 3, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.80±0.20°, 8.41±0.20°, 12.16±0.20°, 12.72±0.20°, 13.48±0.20°, 14.72±0.20°, 15.92±0.20°, 16.53±0.20°, 16.89±0.20°, 17.37±0.20°, 18.12±0.20°, 21.44±0.20°, 24.06±0.20°, 24.84±0.20°, 25.55±0.20°, 27.12±0.20° and 29.03±0.20°.

5. The crystal form A according to claim 4, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 5.80, 8.41, 11.58, 12.16, 12.72, 13.07, 13.48, 14.72, 15.92, 16.53, 16.89, 17.37, 18.12, 18.43, 19.11, 21.24, 21.44, 22.41, 2 3.02, 23.23, 23.52, 24.06, 24.84, 25.34, 25.55, 26.08, 26.49, 26.89, 27.12, 27.92, 28.62, 29.03, 29.75, 30.10, 31.21, 32.10, 32.89, 33.26, 34.45, 35.64, 36.06, 37.24, 38.

84.

6. The crystal form A according to claim 5, whose XRPD pattern is substantially as shown in Figure 1.

7. The crystal form A according to any one of claims 1 to 6, wherein the differential scanning calorimetry (DSC) curve thereof has an endothermic peak starting point at 164.0±5°C.

8. The crystal form A according to claim 7, whose DSC spectrum is shown in Figure 2.

9. The crystal form A according to any one of claims 1 to 6, wherein the thermogravimetric analysis (TGA) curve thereof shows a weight loss of 1.25% at 150.0±3°C.

10. The crystal form A according to claim 9, whose TGA spectrum is shown in Figure 3.

11. The compound of formula (II) has the following structural formula:

12. A crystalline form of compound B of formula (II), having an X-ray powder diffraction pattern (XRPD) with characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 14.59±0.20°, 19.11±0.20°, and 21.75±0.20°; 13. The crystal form B according to claim 12, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 19.11±0.20°, 21.75±0.20°, 24.94±0.20° and 25.73±0.20°.

14. The crystal form B according to claim 13, wherein its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 16.62±0.20°, 19.11±0.20°, 21.75±0.20°, 22.23±0.20°, 24.94±0.20°, 25.73±0.20°, 26.22±0.20°, and 27.88±0.20°.

15. The crystal form B according to claim 14, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.22±0.20°, 7.66±0.20°, 8.11±0.20°, 13.89±0.20°, 14.59±0.20°, 15.31±0.20°, 16.62±0.20°, 17.65±0.20°, 18.35±0.20°, 19.11±0.20°, 21.75±0.20°, 22.23±0.20°, 24.94±0.20°, 25.73±0.20°, 26.22±0.20°, and 27.88±0.20°.

16. The crystal form B according to claim 15, wherein the X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ angles: 6.22, 7.66, 8.11, 11.34, 12.21, 13.89, 14.59, 14.84, 15.31, 16.20, 16.62, 17.44, 17.65, 18.35, 19.11, 19.84, 20.43, 21.75, 22.23, 22.46, 23.50, 24.36, 24.94, 25.73, 26.22, 27.43, 27.88, 28.72, 29.14, 30.11, 30.64, 32.04, 32.87, 33.46, 35.89, 38.78 and 39.

28.

17. The crystal form B according to claim 16, whose XRPD pattern is substantially as shown in Figure 4.

18. The crystal form B according to any one of claims 12 to 17, wherein the differential scanning calorimetry (DSC) curve thereof has an endothermic peak at 153.7±3°C.

19. The crystal form B according to claim 18, whose DSC spectrum is shown in Figure 5.

20. The crystal form B according to any one of claims 12 to 17, wherein the thermogravimetric analysis (TGA) curve thereof shows a weight loss of 2.88% at 150.0±3°C.

21. The crystal form B according to claim 20, whose TGA spectrum is shown in Figure 6.

22. A pharmaceutical composition comprising a therapeutically effective dose of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof as an active ingredient and a pharmaceutically acceptable excipient; 23. The pharmaceutical composition according to claim 22, wherein the dosage form of the pharmaceutical composition is selected from capsules, granules, injections, pills, syrups, powders, ointments, emulsions, solutions, suspensions or tinctures.

24. The pharmaceutical composition according to claim 22, wherein the excipient is selected from excipients, fillers, binders, humectants, disintegrants, solubilizers, absorption accelerators, adsorbents, diluents, solubilizers, emulsifiers, lubricants, wetting agents, glidants, suspending agents, flavoring agents, spices or mixtures thereof.

25. A method for preparing the compound of formula (I), wherein the method is as follows: in, R1 is selected from H, CH3, Boc, Cbz, Bn and PMB; 1) When R1 is selected from H, the preparation method of the compound of formula (I) is selected from: 2) When R1 is selected from CH3, the compound of formula (II-3) is the compound of formula (I); 3) When R1 is selected from Boc, Cbz, Bn and PMB, the preparation method of the compound of formula (I) is selected from: Wherein, the preparation method of the compound of formula (II-3) is selected from: Method 1 includes: Method 2 includes: Method 3 includes: in, R1 is selected from H, CH3, Boc, Cbz, Bn and PMB; R2 is selected from Bn, PMB, PNB and MOM; R4 is selected from CH3, MOM, Bn, SEM and Ac; R5 is selected from F, Cl, Br, I, OTf and OCH2CF3; R 31 Selected from B(OH)2; and BF3K; R6 is selected from F, Cl, Br and I; R7 is selected from Cl, Br, OTf, OCH2CF3 and OTs.

26. A method for preparing a compound of formula (I), It includes the following steps: Step 1: react compound EE-3 to obtain compound EE-4. Step 2: react compound EE-4 to obtain compound EE-5. Step 3: Compound EE-5 and compound AA react to obtain compound H-1. Step 4: reacting compound H-1 to obtain a compound of formula (I), 27. A method for preparing the compound of formula (I) according to claim 26, comprising the steps of: Step 1: react compound EE-3 with reagents A and B to obtain compound EE-4. Step 2: Compound EE-4 is reacted with reagents D, E, F, and G to obtain compound EE-5. Step 3: Compound EE-5 and compound AA react with reagents I and J to obtain compound H-1. Step 4: reacting compound H-1 with reagent K to obtain a compound of formula (I), in, Reagent A is selected from chloromethyl ether, bromomethyl ether, iodomethyl ether and methoxymethyl methanesulfonate, preferably chloromethyl ether; Reagent B is selected from potassium carbonate, cesium carbonate, sodium carbonate, lithium carbonate, sodium hydrogen, lithium hexamethyldisilazide, sodium hexamethyldisilazide, potassium hexamethyldisilazide, potassium tert-butoxide, sodium tert-butoxide, sodium hydroxide, lithium hydroxide and potassium hydroxide, preferably potassium carbonate; Solvent C is selected from DMF, DCM, THF, 2-MeTHF, EtOAc, i-PrOAc, NMP and dioxane, preferably DMF; Reagent D is selected from bis-xanaphthene boronic acid ester and xanaphthene borane, preferably xanaphthene borane; Reagent E is selected from Pd(dppf)Cl2, Pd(dppf)Cl2.CH2Cl2, bis(acetonitrile)palladium(II)dichloride, Pd(OAc)2 and Pd2(dba)3, preferably Pd(OAc)2; Reagent F is selected from Sphos, Xphos, XantPhos and 2-(dicyclohexylphosphino)biphenyl, preferably 2-(dicyclohexylphosphino)biphenyl; Reagent G is selected from potassium acetate, sodium acetate, TEA, DIPEA and potassium 2-ethylhexanoate, preferably TEA; Solvent H is selected from dioxane, DMSO, DMF, NMP, DCM, THF, 2-MeTHF and water, preferably dioxane; Reagent I is selected from Pd(OAc)2-BINAP, Pd2(dba)3-Xphos, Brettphos-Pd-G3, Pd(PPh3)2Cl2, Pd(PPh3)4, Pd(dppf)Cl2CH2Cl2 and Pd(OAc)2-Sphos, preferably Pd(dppf)Cl2CH2Cl2; Reagent J is selected from Cs2CO3, K2CO3, Na2CO3, K3PO4, KOAc, KF and TEA, preferably K2CO3; Reagent K is selected from hydrochloric acid, hydrobromic acid, trifluoroacetic acid, sulfuric acid, formic acid, methanesulfonic acid, benzenesulfonic acid and p-toluenesulfonic acid, preferably hydrochloric acid.

28. A method for preparing an intermediate compound represented by formula (AA): in, R 51 selected from the group consisting of F, Cl, Br and I; R 11 Selected from CH3, Boc, Cbz, Bn and PMB.

29. The method for preparing the intermediate compound of formula (AA) according to claim 28, wherein: R 51 Selected from Cl, R 11 Selected from Boc.

30. A method for preparing a compound of formula (I), wherein the method is selected from the group consisting of: Method 1 includes: Method 2 includes: Method 3 includes: Method 4 includes: Method 5 includes: Method 6 includes: Method seven includes: Method 8 includes: Method nine includes: Method 10 includes: Method 11 includes: Method 12 includes: Method 13 includes: Method 14 includes:

31. Use of the crystalline form A of compound (I) according to any one of claims 1 to 10, the compound of formula (II) according to claim 11, the crystalline form B of compound (II) according to any one of claims 12 to 21, the pharmaceutical composition according to any one of claims 22 to 24, or the preparation method according to any one of claims 25 to 30 in the preparation of drugs for treating diseases associated with NLRP3 inflammasome.

32. The use according to claim 31, wherein the disease associated with NLRP3 inflammasome is selected from NLRP3 inflammasome-associated neuroinflammatory diseases (such as brain inflammation, brain damage) and neurodegenerative diseases (such as Parkinson's disease, Alzheimer's disease, multiple sclerosis).

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