TW202340193A - Crystal of pyrazolopyrimidinone compound and salt thereof - Google Patents

Abstract

The present invention relates to a crystal form of a compound as represented in formula I, various salt types and crystal forms thereof, and further relates to a preparation method for the crystal form of the compound as represented in formula I, various salt types and crystal forms thereof, and a pharmaceutical composition containing the crystal form, various salt types and crystal forms thereof, and the use thereof in the preparation of a drug for treating diseases, disorders or conditions, or a treatment method for treating the diseases, disorders or conditions.

Description

Crystallization of pyrazolopyrimidinone compounds and their salts

The invention belongs to the field of biomedicine and mainly relates to the crystal forms of pyrazolopyrimidinone compounds, various salt forms and their crystal forms, as well as preparation methods of the crystal forms, pharmaceutical compositions and related uses.

Src homology region 2-containing protein tyrosine phosphatase 2 (SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene. PTPN11 was the first to be The discovered proto-oncogene encoding tyrosine phosphokinase (Chan R J et al. PTPN11 is the first identified proto-oncogene that encodes a tyrosine phosphatase. Blood, 2007, 109:862-867), which encodes the SHP2 protein containing N terminal SHP2 domain (N-SHP2), C-terminal SHP2 domain (C-SHP2), protein phosphatase catalytic domain (PTP), two C-terminal tyrosine residues (Y542 and Y580) and a proline-rich Motif of amino acid (Pro).

In recent years, studies have mainly believed that the Ras/ERK pathway is the most important signal transduction pathway in which SHP2 plays a role, and its mechanism (Dance M et al. The molecular functions of Shp2 in the RAS/mitogen-activated protein kinase (ERK1/2) pathway. Cell Signal, 2008, 20:453-459) is roughly as follows: after the growth factor receptor is activated, its tyrosine residue undergoes autophosphorylation to phosphotyrosine for Grb2 and SHP2 (adapter protein containing SH2 domain) Binding region SH2 provides a docking site. The binding of Grb2 to phosphorylated growth factor receptors leads to the accumulation of SOS proteins in the plasma membrane. SOS, as a guanine nucleotide exchange factor (GEF), can catalyze the conversion of the membrane-bound protein Ras from inactive Ras-GDP to active Ras-GTP. Ras-GTP further contacts the downstream signaling system, activating Ser/Thr kinase Raf1, etc., and then activates ERK under the action of the regulatory kinase MEK. After activation, ERK directly acts on target molecules in the cytoplasm or is transferred to the nucleus to regulate genes. Transcription, causing cells to proliferate or differentiate. This process may also be affected by SHP2 binding protein and substrate (SHP substrate-1, SHPS-1), Ras-GTPase activating protein (Ras-GAP), and other Src members.

SHP2 protein not only regulates the Ras/ERK signaling pathway, but also has been reported to regulate multiple signaling pathways such as JAK-STAT3, NF-κB, PI3K/Akt, RHO and NFAT, thereby regulating cell proliferation, differentiation, migration, apoptosis and other physiological processes. learning function.

SHP2 has been shown to be associated with a variety of diseases. Tartaglia et al. (Tartaglia M et al. Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan sydrome. Nat Genet, 2001, 29:465-468) found that about 50% of Noonan sydrome Patients with Nan syndrome are associated with missense mutations in PTPN11. In addition, studies have found that PTPN11 mutations are an important cause of JMMLL and various leukemias (Tartaglia M et al. Nat Genet, 2003, 34: 148-150; Loh ML et al. Blood, 2004, 103:2325-2331; Tartaglia M et al. al. Br J Haematol, 2005, 129:333-339; Xu R et al. Blood, 2005, 106:3142-3149.). With the in-depth research on PTPN11/SHP2, it has been found that it is related to the occurrence of lung cancer, gastric cancer, colon cancer, melanoma, thyroid cancer and other cancers (Tang Chunlan et al., Chinese Journal of Lung Cancer, 2010, 13:98-101; Higuchi M et al. Cancer Sci, 2004, 95:442-447; Bentires-Al j M et al. Cancer Res, 2004, 64:8816-8820; Martinelli S et al. Cancer Genet Cytogenet, 2006, 166:124- 129.).

PCT International Application PCT/CN2021/099275 describes a class of pyrazolopyrimidinone derivatives used as SHP2 protease inhibitors. Most of these compounds can effectively inhibit SHP2. However, there are still unmet needs in terms of treatment options for SHP2-mediated diseases. Here, we further screen the crystal forms and salt forms of pyrazolopyrimidinone derivatives to meet the medical needs of patients.

The present invention The present invention relates to compound ( S )-6-(1-amino-1,3-dihydrospiro[indene-2,4'-piridin]-1'-yl)-3-(1 Crystalline and salt forms of -phenylcyclopropyl)-1,5-dihydro- 4H -pyrazolo[3,4- d ]pyrimidin-4-one. The preparation method and structure of the compound represented by the structural formula I have been described in detail in the PCT patent application PCT/CN2021/099275. Formula I

Salt forms of compounds of formula I

In some embodiments, an acid and a compound of Formula I can form corresponding salts in corresponding systems, and these salt-form compounds can exist in various physical forms. For example, it can be in solution, suspension or solid form. In certain embodiments, salt-form compounds are in solid form. In solid form, the compound may be amorphous, crystalline, or a mixture thereof. Preferably, the salt form of the compound of formula I is a salt formed by the compound of formula I and the following acids: methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, L-lactic acid, L-tartaric acid, fumaric acid, L-apple Acid and hydrochloride; more preferably, the acid is methanesulfonic acid. Further, various salts of the compounds of formula I may exist in the form of crystalline forms.

Exemplary examples of different crystal forms of the various salts of the compounds of formula I are as follows.

Form A of methanesulfonate salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the mesylate crystal form A has the characteristics of diffraction angles 2θ of 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, and 23.5°±0.2°. peak.

Preferably, the X-ray powder diffraction pattern of the mesylate crystal form A has diffraction angles 2θ of 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, 16.9°±0.2°, 23.5 °±0.2° characteristic peak.

Preferably, the X-ray powder diffraction pattern of the methanesulfonate crystal form A has diffraction angles 2θ of 5.6°±0.2°, 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, and 16.9 The characteristic peaks are °±0.2°, 19.8°±0.2°, 20.6°±0.2°, 22.5°±0.2°, and 23.5°±0.2°.

More preferably, the main data of the X-ray powder diffraction pattern of the mesylate crystal form A is shown in Table 1.

Table 1 No. Diffraction angle 2θ interplanar spacing relative strength 1 5.6° 15.81Å 8.00% 2 7.2° 12.34Å 26.60% 3 10.0° 8.87Å 100.00% 4 14.4° 6.12Å 28.50% 5 14.5° 6.10Å 38.00% 6 16.9° 5.26Å 27.10% 7 19.7° 4.51Å 18.50% 8 19.8° 4.48Å 25.30% 9 20.0° 4.43Å 16.10% 10 20.6° 4.30Å 9.70% 11 22.5° 3.94Å 10.50% 12 23.5° 3.79Å 54.50% 13 24.9° 3.58Å 13.30%

Preferably, the methanesulfonate crystalline Form A has an X-ray powder diffraction pattern substantially as shown in Figure 1.

Further, the methanesulfonate crystal form A has a differential scanning calorimetry (DSC) pattern substantially as shown in Figure 2.

Further, the methanesulfonate crystal form A has a thermogravimetric analysis (TGA) pattern substantially as shown in Figure 3.

Further, the methanesulfonate crystal form A has a ¹ H-NMR pattern substantially as shown in Figure 4.

Furthermore, the mesylate crystal form A is an anhydrous form.

Form D of the mesylate salt of the compound of formula I.

Preferably, the X-ray powder diffraction pattern of the mesylate crystal form D has a diffraction angle 2θ of 5.7°±0.2°, 16.4°±0.2°, 19.3°±0.2°, and 20.1°±0.2°. The characteristic peak is 22.9°±0.2°.

More preferably, the X-ray powder diffraction pattern of the mesylate crystal form D has a diffraction angle 2θ of 5.7°±0.2°, 11.9°±0.2°, 16.4°±0.2°, and 17.3°±0.2°. The characteristic peaks are 19.3°±0.2°, 20.1°±0.2°, 22.9°±0.2°, and 26.1°±0.2°.

Preferably, the methanesulfonate crystal form D has an X-ray powder diffraction pattern substantially as shown in Figure 6.

Further, the methanesulfonate crystal form D has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 7.

Further, the methanesulfonate crystal form D has a 1H-NMR pattern basically as shown in Figure 8.

Further, the single crystal data of the methanesulfonate crystal form D is shown in Table 9, the three-dimensional structure diagram of its asymmetric unit is shown in Figure 5, and its unit cell stacking diagram is shown in Figure 5-1.

Furthermore, the mesylate crystal form D is an anhydrous form.

Compound of formula I benzene sulfonate crystal form A

Preferably, the X-ray powder diffraction pattern of the benzene sulfonate crystal form A has a diffraction angle 2θ of 8.4°±0.2°, 11.0°±0.2°, 14.2°±0.2°, and 16.8°±0.2°. The characteristic peaks are 17.4°±0.2°, 18.8°±0.2°, and 25.6°±0.2°.

More preferably, the X-ray powder diffraction pattern of the benzene sulfonate crystal form A has a diffraction angle 2θ of 6.2°±0.2°, 8.4°±0.2°, 11.0°±0.2°, 11.8°±0.2°, The characteristic peaks are 13.8°±0.2°, 14.2°±0.2°, 15.9°±0.2°, 16.8°±0.2°, 17.4°±0.2°, 18.8°±0.2°, and 25.6°±0.2°.

Preferably, the benzene sulfonate crystalline Form A has an X-ray powder diffraction pattern substantially as shown in Figure 9.

Further, the benzene sulfonate crystal form A has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 10.

Further, the benzene sulfonate crystal form A has a 1H-NMR spectrum substantially as shown in Figure 11.

Compound of formula I p-toluenesulfonate crystal form A

Preferably, the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A has a diffraction angle 2θ of 6.0°±0.2°, 10.4°±0.2°, 14.2°±0.2°, and 18.7°±0.2°. , a characteristic peak of 22.3°±0.2°.

More preferably, the X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A has a diffraction angle 2θ of 6.0°±0.2°, 10.4°±0.2°, 11.2°±0.2°, and 12.4°±0.2°. , 13.0°±0.2°, 14.2°±0.2°, 18.3°±0.2°, 18.7°±0.2°, and 22.3°±0.2° characteristic peaks.

Preferably, the p-toluenesulfonate crystal form A has an X-ray powder diffraction pattern substantially as shown in Figure 12.

Further, the p-toluenesulfonate crystal form A has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 13.

Further, the p-toluenesulfonate crystal form A has a 1H-NMR spectrum basically as shown in Figure 14.

Compound of formula I L-lactate crystalline form A

Preferably, the X-ray powder diffraction pattern of the L-lactate crystal form A has a diffraction angle 2θ of 6.2°±0.2°, 6.7°±0.2°, 12.2°±0.2°, and 13.9°±0.2°. The characteristic peaks are 18.5°±0.2°, 21.0°±0.2°, and 25.6°±0.2°.

More preferably, the X-ray powder diffraction pattern of the L-lactate crystal form A has a diffraction angle 2θ of 6.2°±0.2°, 6.7°±0.2°, 12.2°±0.2°, and 13.9°±0.2°, Characteristic peaks at 17.2°±0.2°, 17.9°±0.2°, 18.5°±0.2°, 21.0°±0.2°, 25.1°±0.2°, and 25.6°±0.2°;

Preferably, the L-lactate crystal form A has an X-ray powder diffraction pattern substantially as shown in Figure 15.

Further, the L-lactate crystal form A has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 16.

Further, the L-lactate crystalline form A has a 1H-NMR spectrum substantially as shown in Figure 17.

Form A of compound L-tartrate of formula I

Preferably, the X-ray powder diffraction pattern of the L-tartrate crystal form A has a diffraction angle 2θ of 5.9°±0.2°, 12.6°±0.2°, 15.8°±0.2°, and 18.9°±0.2°. The characteristic peak is 24.6°±0.2°.

More preferably, the X-ray powder diffraction pattern of the L-tartrate crystal form A has a diffraction angle 2θ of 5.9°±0.2°, 10.8°±0.2°, 12.6°±0.2°, and 15.1°±0.2°, The characteristic peaks are 15.8°±0.2°, 17.9°±0.2°, 18.6°±0.2°, 18.9°±0.2°, 19.4°±0.2°, 24.6°±0.2°, and 25.3°±0.2°.

Preferably, the L-tartrate crystal form A has an X-ray powder diffraction pattern substantially as shown in Figure 18.

Further, the L-tartrate crystal form A has a thermogravimetric analysis pattern basically as shown in Figure 19.

Further, the L-tartrate crystalline form A has a 1H-NMR spectrum substantially as shown in Figure 20.

Form A of fumarate salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the fumarate crystal form A has a diffraction angle 2θ of 7.5°±0.2°, 15.0°±0.2°, 16.2°±0.2°, and 17.3°±0.2°. The characteristic peaks are 22.8°±0.2° and 28.7°±0.2°.

More preferably, the X-ray powder diffraction pattern of the fumarate crystal form A has a diffraction angle 2θ of 7.5°±0.2°, 15.0°±0.2°, 16.2°±0.2°, and 17.3°±0.2°. The characteristic peaks are 22.8°±0.2°, 28.7°±0.2°, and 29.3°±0.2°.

Preferably, the fumarate salt Form A has an X-ray powder diffraction pattern substantially as shown in Figure 21.

Further, the fumarate crystal form A has a thermogravimetric analysis pattern substantially as shown in Figure 22.

Further, the fumarate crystalline Form A has a 1H-NMR spectrum substantially as shown in Figure 23.

Form A of compound L-malate of formula I

Preferably, the X-ray powder diffraction pattern of the L-malate crystal form A has a diffraction angle 2θ of 5.1°±0.2°, 10.2°±0.2°, 17.2°±0.2°, and 20.6°±0.2°. , a characteristic peak of 23.0°±0.2°.

More preferably, the X-ray powder diffraction pattern of the L-malate crystal form A has a diffraction angle 2θ of 5.1°±0.2°, 6.1°±0.2°, 10.2°±0.2°, and 16.2°±0.2°. , 17.2°±0.2°, 20.6°±0.2°, and 23.0°±0.2° characteristic peaks.

Preferably, the L-malate crystal form A has an X-ray powder diffraction pattern substantially as shown in Figure 24.

Further, the L-malate crystal form A has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 25.

Further, the L-malate crystalline form A has a 1H-NMR spectrum substantially as shown in Figure 26.

Form A of the hydrochloride salt of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form A has a diffraction angle 2θ of 6.9°±0.2°, 10.4°±0.2°, 16.2°±0.2°, 16.9°±0.2°, and 18.1 The characteristic peaks are °±0.2°, 18.6°±0.2°, and 19.5°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form A has a diffraction angle 2θ of 6.9°±0.2°, 10.4°±0.2°, 13.9°±0.2°, 14.2°±0.2°, 15.5 The characteristic peaks are °±0.2°, 16.2°±0.2°, 16.9°±0.2°, 17.5°±0.2°, 18.1°±0.2°, 18.6°±0.2°, and 19.5°±0.2°.

Preferably, the hydrochloride crystal form A has an X-ray powder diffraction pattern substantially as shown in Figure 27.

Further, the hydrochloride crystal form A has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 28.

Form B of the hydrochloride salt of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form B has a diffraction angle 2θ of 7.3°±0.2°, 10.0°±0.2°, 10.5°±0.2°, 16.0°±0.2°, and 18.7 °±0.2°, 20.8°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form B has a diffraction angle 2θ of 7.3°±0.2°, 10.0°±0.2°, 10.5°±0.2°, 14.0°±0.2°, 14.6 The characteristic peaks are °±0.2°, 16.0°±0.2°, 18.7°±0.2°, 19.2°±0.2°, 19.6°±0.2°, 20.8°±0.2°, and 22.8°±0.2°.

Preferably, the hydrochloride crystal form B has an X-ray powder diffraction pattern substantially as shown in Figure 29.

Further, the hydrochloride crystal form B has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 30.

Form D of the hydrochloride salt of the compound of formula I:

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form D has a diffraction angle 2θ of 7.6°±0.2°, 9.1°±0.2°, 10.7°±0.2°, 17.8°±0.2°, and 18.8 °±0.2° characteristic peak.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form D has a diffraction angle 2θ of 7.6°±0.2°, 9.1°±0.2°, 10.7°±0.2°, 17.8°±0.2°, and 18.8 The characteristic peaks are °±0.2°, 19.2°±0.2°, 19.5°±0.2°, and 19.9°±0.2°.

Preferably, the hydrochloride crystal form D has an X-ray powder diffraction pattern substantially as shown in Figure 31.

Further, the hydrochloride crystal form D has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 32.

Form E of the hydrochloride salt of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form E has diffraction angles 2θ of 10.7°±0.2°, 12.3°±0.2°, 15.1°±0.2°, 15.6°±0.2°, and 19.8 The characteristic peaks are °±0.2°, 20.6°±0.2°, 21.5°±0.2°, 22.7°±0.2°, and 23.7°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form E has a diffraction angle 2θ of 8.3°±0.2°, 10.7°±0.2°, 12.3°±0.2°, 15.1°±0.2°, 15.6 The characteristic peaks are °±0.2°, 17.3°±0.2°, 19.3°±0.2°, 19.8°±0.2°, 20.6°±0.2°, 21.5°±0.2°, 22.7°±0.2°, and 23.7°±0.2°.

Preferably, the hydrochloride crystal form E has an X-ray powder diffraction pattern substantially as shown in Figure 33.

Further, the hydrochloride crystal form E has a thermogravimetric analysis diagram substantially as shown in Figure 34.

Form F of the hydrochloride salt of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form F has a diffraction angle 2θ of 6.9°±0.2°, 7.6°±0.2°, 12.8°±0.2°, 15.3°±0.2°, and 20.0 °±0.2° characteristic peak.

Preferably, the hydrochloride crystal form F has an X-ray powder diffraction pattern substantially as shown in Figure 35.

Compound of formula I hydrochloride crystalline form G

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form G has a diffraction angle 2θ of 7.2°±0.2°, 9.6°±0.2°, 10.1°±0.2°, 15.5°±0.2°, and 18.8 The characteristic peaks are °±0.2°, 19.3°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 21.7°±0.2°, and 22.2°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form G has a diffraction angle 2θ of 7.2°±0.2°, 9.6°±0.2°, 10.1°±0.2°, 10.7°±0.2°, and 11.0 The characteristic peaks are °±0.2°, 15.5°±0.2°, 18.8°±0.2°, 19.3°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 21.7°±0.2°, and 22.2°±0.2°.

Preferably, the hydrochloride crystal form G has an X-ray powder diffraction pattern substantially as shown in Figure 36.

Further, the hydrochloride crystal form G has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 37.

Form H of the hydrochloride salt of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form H has a diffraction angle 2θ of 7.1°±0.2°, 9.8°±0.2°, 10.5°±0.2°, 11.1°±0.2°, and 15.6 The characteristic peaks are °±0.2°, 18.8°±0.2°, 20.3°±0.2°, and 22.4°±0.2°.

Preferably, the hydrochloride crystal form H has an X-ray powder diffraction pattern substantially as shown in Figure 38.

Further, the hydrochloride crystal form H has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 39.

Compound of formula I hydrochloride salt form I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form I has a diffraction angle 2θ of 7.5°±0.2°, 11.8°±0.2°, 16.7°±0.2°, 18.6°±0.2°, and 18.9 °±0.2°, 21.8°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form I has a diffraction angle 2θ of 7.5°±0.2°, 10.7°±0.2°, 11.8°±0.2°, 16.7°±0.2°, and 18.6 The characteristic peaks are °±0.2°, 18.9°±0.2°, and 21.8°±0.2°.

Preferably, the hydrochloride salt Form I has an X-ray powder diffraction pattern substantially as shown in Figure 40.

Further, the hydrochloride crystal form I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 41.

Form J of hydrochloride salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form J has a diffraction angle 2θ of 7.0°±0.2°, 10.5°±0.2°, 14.1°±0.2°, 17.6°±0.2°, and 19.0 The characteristic peaks are °±0.2°, 24.9°±0.2°, and 26.1°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form J has a diffraction angle 2θ of 7.0°±0.2°, 10.5°±0.2°, 14.1°±0.2°, 17.6°±0.2°, and 19.0 The characteristic peaks are °±0.2°, 21.4°±0.2°, 24.9°±0.2°, 26.1°±0.2°, and 28.2°±0.2°.

Preferably, the hydrochloride crystal Form J has an X-ray powder diffraction pattern substantially as shown in Figure 42.

Further, the hydrochloride crystal form J has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 43.

Form K of hydrochloride salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form K has diffraction angles 2θ of 6.9°±0.2°, 7.6°±0.2°, 11.4°±0.2°, 17.9°±0.2°, and 18.8 °±0.2°, 19.6°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form K has a diffraction angle 2θ of 6.9°±0.2°, 7.6°±0.2°, 11.4°±0.2°, 12.9°±0.2°, and 14.5 Characteristic peaks of °±0.2°, 16.0°±0.2°, 17.6°±0.2°, 17.9°±0.2°, 18.8°±0.2°, and 19.6°±0.2°;

Preferably, the hydrochloride crystal form K has an X-ray powder diffraction pattern substantially as shown in Figure 44.

Further, the hydrochloride crystal form K has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 45.

Compound of formula I hydrochloride crystalline form L

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form L has a diffraction angle 2θ of 6.1°±0.2°, 11.4°±0.2°, 15.0°±0.2°, 15.4°±0.2°, and 18.9 °±0.2° characteristic peak.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form L has a diffraction angle 2θ of 6.1°±0.2°, 11.4°±0.2°, 15.0°±0.2°, 15.4°±0.2°, and 18.9 The characteristic peaks are °±0.2°, 21.4°±0.2°, 22.3°±0.2°, 23.0°±0.2°, and 23.8°±0.2°.

Preferably, the hydrochloride crystal form L has an X-ray powder diffraction pattern substantially as shown in Figure 46.

Further, the hydrochloride crystal form L has a thermogravimetric analysis diagram substantially as shown in Figure 47.

Form M of hydrochloride salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form M has a diffraction angle 2θ of 7.1°±0.2°, 9.7°±0.2°, 10.4°±0.2°, 14.2°±0.2°, and 15.4 The characteristic peaks are °±0.2°, 19.4°±0.2°, 20.1°±0.2°, 21.4°±0.2°, and 22.1°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form M has a diffraction angle 2θ of 7.1°±0.2°, 9.7°±0.2°, 10.4°±0.2°, 11.0°±0.2°, 14.2 The characteristic peaks are °±0.2°, 15.4°±0.2°, 19.4°±0.2°, 20.1°±0.2°, 21.4°±0.2°, 22.1°±0.2°, and 26.6°±0.2°.

Preferably, the hydrochloride crystal form M has an X-ray powder diffraction pattern substantially as shown in Figure 48.

Further, the hydrochloride crystal form M has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 49.

Compound of formula I hydrochloride crystalline form N

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form N has a diffraction angle 2θ of 6.4°±0.2°, 11.1°±0.2°, 12.8°±0.2°, 15.7°±0.2°, 16.2 The characteristic peaks are °±0.2°, 16.6°±0.2°, 18.7°±0.2°, 20.7°±0.2°, 22.1°±0.2°, 23.1°±0.2°, and 23.9°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form N has a diffraction angle 2θ of 6.4°±0.2°, 11.1°±0.2°, 11.4°±0.2°, 11.9°±0.2°, and 12.8 °±0.2°, 15.7°±0.2°, 16.2°±0.2°, 16.6°±0.2°, 18.7°±0.2°, 19.2°±0.2°, 20.7°±0.2°, 22.1°±0.2°, 23.1°± Characteristic peaks at 0.2° and 23.9°±0.2°.

Preferably, the hydrochloride crystal form N has an X-ray powder diffraction pattern substantially as shown in Figure 50.

Further, the hydrochloride crystal form N has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 51.

Compound of Formula I Hydrochloride Form O

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form O has a diffraction angle 2θ of 8.0°±0.2°, 10.6°±0.2°, 12.1°±0.2°, 12.3°±0.2°, and 14.8 The characteristic peaks are °±0.2°, 15.5°±0.2°, 16.6°±0.2°, 19.3°±0.2°, 20.5°±0.2°, 22.5°±0.2°, and 23.9°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form O has a diffraction angle 2θ of 8.0°±0.2°, 10.6°±0.2°, 12.1°±0.2°, 12.3°±0.2°, and 13.6 °±0.2°, 14.8°±0.2°, 15.5°±0.2°, 16.6°±0.2°, 19.3°±0.2°, 20.5°±0.2°, 21.3°±0.2°, 22.5°±0.2°, 23.9°± Characteristic peaks at 0.2° and 25.9°±0.2°.

Preferably, the hydrochloride salt Form O has an X-ray powder diffraction pattern substantially as shown in Figure 52.

Further, the hydrochloride crystal form O has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 53.

Compound of formula I hydrochloride crystalline form P

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form P has a diffraction angle 2θ of 7.3°±0.2°, 9.1°±0.2°, 14.7°±0.2°, 15.6°±0.2°, 19.3 °±0.2°, 21.4°±0.2° characteristic peaks.

Preferably, the hydrochloride crystal form P has an X-ray powder diffraction pattern substantially as shown in Figure 54.

Further, the hydrochloride crystal form P has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 55.

Form Q of hydrochloride salt of compound of formula I

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form Q has a diffraction angle 2θ of 8.5°±0.2°, 10.9°±0.2°, 12.4°±0.2°, 15.3°±0.2°, 15.9 The characteristic peaks are °±0.2°, 19.6°±0.2°, 20.1°±0.2°, 20.6°±0.2°, 21.6°±0.2°, 22.9°±0.2°, and 24.0°±0.2°.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form Q has a diffraction angle 2θ of 8.5°±0.2°, 10.9°±0.2°, 12.4°±0.2°, 15.3°±0.2°, 15.9 °±0.2°, 16.6°±0.2°, 17.6°±0.2°, 18.6°±0.2°, 19.6°±0.2°, 20.1°±0.2°, 20.6°±0.2°, 21.6°±0.2°, 22.9°± Characteristic peaks at 0.2° and 24.0°±0.2°.

Preferably, the hydrochloride crystal form Q has an X-ray powder diffraction pattern substantially as shown in Figure 56.

Further, the hydrochloride crystal form Q has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 57.

Compound of formula I hydrochloride crystal form R

Preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form R has a diffraction angle 2θ of 6.7°±0.2°, 7.8°±0.2°, 12.9°±0.2°, 15.7°±0.2°, and 18.8 °±0.2°, 19.1°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the hydrochloride crystal form R has a diffraction angle 2θ of 6.7°±0.2°, 7.8°±0.2°, 9.6°±0.2°, 10.9°±0.2°, and 12.9 The characteristic peaks are °±0.2°, 15.7°±0.2°, 18.8°±0.2°, 19.1°±0.2°, 19.5°±0.2°, and 20.2°±0.2°.

Preferably, the hydrochloride crystal form R has an X-ray powder diffraction pattern substantially as shown in Figure 58.

Exemplary examples of different crystal forms of the compound of formula I are as follows:

Form A of the compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form A of the compound of formula I has a diffraction angle 2θ of 6.4°±0.2°, 15.5°±0.2°, 16.3°±0.2°, 18.0°±0.2°, 18.7 °±0.2°, 23.5°±0.2° characteristic peaks.

Preferably, the crystal form A of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 59.

Further, the crystal form A of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 60.

Form B of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form B of the compound of formula I has a diffraction angle 2θ of 5.3°±0.2°, 6.5°±0.2°, 7.7°±0.2°, 13.3°±0.2°, and 14.9 The characteristic peaks are °±0.2°, 16.2°±0.2°, 19.9°±0.2°, 26.1°±0.2°, and 29.4°±0.2°.

Preferably, the crystal form B of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 61.

Further, the crystal form B of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 62.

Form C of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form C of the compound of formula I has a diffraction angle 2θ of 5.9°±0.2°, 6.7°±0.2°, 14.1°±0.2°, 15.1°±0.2°, and 17.5 The characteristic peaks are °±0.2°, 18.0°±0.2°, and 20.5°±0.2°.

Preferably, the crystal form C of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 63.

Further, the crystal form C of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 64.

Crystal form D of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form D of the compound of formula I has a diffraction angle 2θ of 5.5°±0.2°, 14.7°±0.2°, 15.4°±0.2°, 16.5°±0.2°, 17.5 °±0.2°, 21.7°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the crystal form D of the compound of formula I has a diffraction angle 2θ of 5.5°±0.2°, 8.0°±0.2°, 14.2°±0.2°, 14.7°±0.2°, and 15.4 The characteristic peaks are °±0.2°, 16.5°±0.2°, 17.5°±0.2°, 18.0°±0.2°, 19.7°±0.2°, and 21.7°±0.2°.

Preferably, the crystal form D of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 65.

Further, the crystal form D of the compound of formula I has a thermogravimetric analysis diagram substantially as shown in Figure 66.

Crystal form E of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form E of the compound of formula I has a diffraction angle 2θ of 5.3°±0.2°, 7.4°±0.2°, 8.9°±0.2°, 12.9°±0.2°, and 14.7 The characteristic peaks are °±0.2°, 17.2°±0.2°, 17.9°±0.2°, 21.3°±0.2°, and 21.8°±0.2°.

Preferably, the crystal form E of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 67.

Further, the crystal form E of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 68.

Form F of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form F of the compound of formula I has a diffraction angle 2θ of 6.6°±0.2°, 15.9°±0.2°, 16.2°±0.2°, 17.7°±0.2°, 21.6 The characteristic peaks are °±0.2°, 23.4°±0.2°, and 29.5°±0.2°.

Preferably, the crystal form F of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 69.

Further, the crystal form F of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 70.

Form G of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form G of the compound of formula I has a diffraction angle 2θ of 5.8°±0.2°, 8.0°±0.2°, 14.6°±0.2°, 15.2°±0.2°, and 15.9 The characteristic peaks are °±0.2°, 17.2°±0.2°, and 17.9°±0.2°.

Preferably, the crystal form G of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 71.

Further, the crystal form G of the compound of formula I has a thermogravimetric analysis diagram substantially as shown in Figure 72.

Crystal form H of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form H of the compound of formula I has a diffraction angle 2θ of 5.8°±0.2°, 13.0°±0.2°, 14.3°±0.2°, 14.8°±0.2°, 16.3 The characteristic peaks are °±0.2°, 17.3°±0.2°, 17.9°±0.2°, and 21.9°±0.2°.

More preferably, the X-ray powder diffraction pattern of the crystal form H of the compound of formula I has a diffraction angle 2θ of 5.8°±0.2°, 7.4°±0.2°, 13.0°±0.2°, 14.3°±0.2°, and 14.8 The characteristic peaks are °±0.2°, 16.3°±0.2°, 17.3°±0.2°, 17.9°±0.2°, 21.9°±0.2°, and 22.5°±0.2°.

Preferably, the crystal form H of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 73.

Further, the crystal form H of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 74.

Crystal form I of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form I of the compound of formula I has a diffraction angle 2θ of 4.0°±0.2°, 7.5°±0.2°, 10.1°±0.2°, 14.7°±0.2°, and 15.1 The characteristic peaks are °±0.2°, 17.8°±0.2°, and 18.9°±0.2°.

Preferably, the crystal form I of the compound of formula I has an X-ray powder diffraction pattern substantially as shown in Figure 75.

Further, the crystal form I of the compound of formula I has a thermogravimetric analysis diagram substantially as shown in Figure 76.

Crystal form J of compound of formula I

Preferably, the X-ray powder diffraction pattern of the crystal form J of the compound of formula I has a diffraction angle 2θ of 6.4°±0.2°, 8.0°±0.2°, 14.7°±0.2°, 16.0°±0.2°, and 17.5 °±0.2°, 22.4°±0.2° characteristic peaks.

More preferably, the X-ray powder diffraction pattern of the crystal form J of the compound of formula I has a diffraction angle 2θ of 6.4°±0.2°, 8.0°±0.2°, 14.7°±0.2°, 16.0°±0.2°, 16.7 The characteristic peaks are °±0.2°, 17.5°±0.2°, 20.5°±0.2°, and 22.4°±0.2°.

Preferably, the compound of Formula I, Form J, has an X-ray powder diffraction pattern substantially as shown in Figure 77.

Further, the crystal form J of the compound of formula I has a differential scanning calorimetry chart and a thermogravimetric analysis chart basically as shown in Figure 78.

The present invention further provides an amorphous compound of the formula I, the X-ray powder diffraction pattern of which has no obvious sharp diffraction peak.

Preferably, its X-ray powder diffraction pattern is basically as shown in Figure 79.

The invention further provides intermediates of compound S2 or S3 or S21 or S22:

The present invention further provides a method for preparing compound S2 or S3 or S21 or S22 or the compound of formula I or the mesylate crystal form A of the compound of formula I, which includes the following steps:

Compound S2 is prepared from compound S1:

Compound S1 is prepared under the action of potassium acetate, water and acetic acid to obtain compound S2; and/or

Compound S3 is prepared from compound S2:

Compound S2 is prepared under the action of the oxidant potassium hydrogen peroxysulfate complex salt to prepare compound S3.

Further, compound S2 is first dissolved in 1,4-dioxane solution and then reacted with potassium hydrogen peroxysulfate complex salt.

Further, the potassium hydrogen peroxysulfate complex salt participates in the reaction in the form of a mixed solution.

Further, the mixed solution of potassium hydrogen peroxysulfate complex salt is prepared by adding the potassium peroxyhydrogen peroxysulfate complex salt to water; and/or

Compound S21 is prepared from compound S3;

Compound S21 is prepared using compound S3 and DHP as raw materials in the presence of tetrahydrofuran and p-toluenesulfonic acid; and/or

Compound S22 is prepared from compound S21:

Compound S22 is prepared by reacting compound S21 and compound S04 as raw materials in the presence of acetonitrile and DIPEA; and/or

Compounds of formula I are prepared from compound S22:

Compound S22 is first treated with hydrogen chloride-1,4-dioxane solution and then treated with sodium bicarbonate aqueous solution to prepare the compound of formula I.

Further, the step 1 is carried out in the presence of dichloromethane and methanol.

Further, the step 2 is carried out in the presence of methanol; and/or

The mesylate crystal form A of the compound of formula I is prepared from the compound of formula I:

Using tetrahydrofuran/water as the solvent, the compound of formula I and methanesulfonic acid are stirred at a temperature of 40 to 80° C. for 8-24 hours. After filtering and drying, the compound of formula I methanesulfonate crystal form A is prepared.

Further, the volume ratio of tetrahydrofuran/water is 10-30:1. Preferably, the volume ratio is 15-25:1. More preferably, the volume ratio is 19:1.

Further, the temperature is 45-55°C. More preferably, the temperature is 50°C.

Further, the material molar ratio of the compound of formula I and methanesulfonic acid: 0.8-1.2: Preferably, the material molar ratio is 0.9-1.1; more preferably, the material molar ratio is 1-1.

Further, the compound of formula I is first added to the solvent, stirred to dissolve, and then methanesulfonic acid is added; the stirring and dissolving is performed at 40-60°C.

The present invention further provides a method for preparing the mesylate crystal form D of the compound of formula I, which is prepared from the compound of formula I:

Using ethanol/water as a solvent, the compound of formula I and methanesulfonic acid react at a temperature of 20 to 60° C. for 0.5-3 hours. After filtration and drying, the mesylate crystal form D of the compound of formula I is prepared.

Further, the volume ratio of ethanol/water is 10-30:1; preferably, the volume ratio is 15-25:1; more preferably, the volume ratio is 19:1.

Further, the temperature is 30-50°C; more preferably, the temperature is 40°C.

Further, the material molar ratio of the compound of formula I and methanesulfonic acid: 0.8-1.2; preferably, the material molar ratio is 0.9-1.1; more preferably, the material molar ratio is 1-1.

Further, the compound of formula I and methanesulfonic acid are stirred after reacting, the stirring time is 1-24 hours, and the stirring temperature is 15-30°C; preferably, the stirring time is 4 hours, and the stirring temperature is room temperature.

The present invention further provides another preparation method of the mesylate crystal form A of the compound of formula I, which is prepared by the following method:

j Use tetrahydrofuran/water as the solvent, stir and dissolve the compound of formula I at a temperature of 40 to 80°C, and add methanesulfonic acid with an equivalent of 0.1-0.3 relative to the compound of formula I.

k While maintaining the reaction temperature, add the seeds of methanesulfonic acid crystal form D prepared above, continue to add methanesulfonic acid with an equivalent of 0.7-0.9 relative to the compound of formula I, lower the temperature, react for 1-12 hours, filter, and obtain Crystal form D.

l Mix the obtained crystal form D and the solvent at 40 to 80°C, then add the seed crystals of the crystal form A prepared above, and keep the reaction until all the solids are converted into crystal form A; cool down and filter to prepare crystal form A.

Further, in step j, the volume ratio of tetrahydrofuran/water is 10-30:1; preferably, the volume ratio is 15-25:1; more preferably, the volume ratio is 19:1.

Further, in step j, the stirring temperature is 45 to 55°C; more preferably, the temperature is 50°C.

Further, in step j, the feeding amount of methanesulfonic acid is 0.1-0.3 equivalents relative to the compound of formula I; in step k, the feeding amount of methanesulfonic acid is 0.7-0.9 equivalents relative to the compound of formula I.

Further, in step 1, the solvent is ethyl acetate and water; preferably, the volume ratio of ethyl acetate and water in the solvent is 10-20:1; more preferably, the volume ratio of ethyl acetate in the solvent is 10-20:1. The volume ratio of ester and water is 13-18:1; more preferably, the order of adding the solvent is to add ethyl acetate first, and then add water.

According to the present invention, the crystal purity of the crystal form of the compound of formula I and its salt form is preferably greater than 50%, such as more than 85%, more than 95%, more than 99% or more than 99.5%.

The present invention further provides a pharmaceutical composition, which contains a therapeutically effective amount of the crystal form, each salt form or its corresponding crystal form of the compound of formula I according to the present invention, and pharmaceutically acceptable excipients, auxiliaries or carriers . In the above pharmaceutical composition, the weight ratio range of the crystal form, each salt form or its corresponding crystal form of the compound of formula I and the excipient, auxiliary agent or carrier is 0.0001 to 10.

Secondly, the present invention also provides preferred embodiments of the above pharmaceutical composition.

Preferably, the above pharmaceutical composition contains a therapeutically effective amount of the crystal form, each salt form or its corresponding crystal form of the compound of formula I according to the present invention, combined with at least one other active ingredient.

Preferably, the pharmaceutical composition is for oral administration.

Preferably, the pharmaceutical composition is in the form of tablets or capsules.

Preferably, the pharmaceutical composition contains 0.01% by weight to 99% by weight of the crystal form, each salt form or the corresponding crystal form of the compound of formula I according to the present invention.

Preferably, the pharmaceutical composition contains 0.05% by weight to 50% by weight of the crystal form, each salt form or the corresponding crystal form of the compound of formula I according to the present invention.

Preferably, the pharmaceutical composition contains 0.1% by weight to 30% by weight of the crystal form, each salt form or the corresponding crystal form of the compound of formula I according to the present invention.

The present invention further provides the use of the crystal form, each salt form or its corresponding crystal form or pharmaceutical composition of the compound of formula I in the preparation of medicines.

The present invention further provides a better technical solution for the application:

Preferably, the application is to treat, prevent, delay or prevent the occurrence or progression of cancer or cancer metastasis.

Preferably, the application is the preparation of drugs for treating or preventing diseases mediated by SHP2.

Preferably, the disease is cancer.

Preferably, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, and esophageal cancer. , lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma and/or pancreatic cancer.

Preferably, the application is as a SHP2 inhibitor.

The present invention also provides a method of applying a therapeutically effective amount of at least any crystal form, each salt form or its corresponding crystal form or pharmaceutical composition of the compound of formula I according to the present invention on a therapeutic object to treat and/or prevent SHP2 mediated disease approaches.

Preferably, in the above method, the disease mediated by SHP2 is cancer.

Preferably, in the above method, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, Breast, esophageal, lung, colon, head, stomach, lymphoma, glioblastoma, and/or pancreatic cancer.

The present invention also provides a method for treating cancer, which includes administering a therapeutically effective amount of at least any crystal form, salt form or corresponding crystal form or pharmaceutical composition of the compound of formula I according to the present invention to the treatment object, so The cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal cancer, lung cancer, and colon cancer , head cancer, stomach cancer, lymphoma, glioblastoma and/or pancreatic cancer.

Preferably, in the above method, the treatment object is a human being.

In the present invention, the terms "about" and "substantially" are used in "having an X-ray powder diffraction pattern approximately as shown in Figure 1" or "having an X-ray powder diffraction pattern substantially as shown in Figure 1" The precise positions of the peaks in the figures are represented and should not be interpreted as absolute values. Because those skilled in the art know that the 2θ value of the X-ray powder diffraction pattern may produce errors due to different measurement conditions (such as the equipment and instruments used) and different samples, the diffraction angle of the X-ray powder diffraction pattern The measurement error is 5% or less. Generally, a difference of ±0.2° from the given value will be considered appropriate. It should also be understood that the relative intensity of the peaks may fluctuate with experimental conditions and sample preparation such as the preferred orientation of the particles in the sample. The use of automatic or fixed divergence slits will also affect the relative intensity calculation. The intensities shown in the X-ray powder diffraction patterns included here are exemplary only and should not be used as an absolute comparison.

Those skilled in the art will appreciate that small variations in data measured by DSC may occur due to changes in sample purity, sample preparation, and measurement conditions (eg, heating rate). It is to be understood that alternative melting point readings may be given by other kinds of instruments or by the use of conditions different from those described below. Therefore, the endotherms quoted in this application are not intended as absolute values, and such measurement errors will be taken into account when interpreting DSC data.

The present invention will be further illustrated by the various embodiments and experimental examples given below, but these embodiments and experimental examples shall not constitute any limitation on the scope of protection claimed by the present invention. In the specific embodiments of the present invention, unless otherwise specified, the techniques or methods described are conventional techniques or methods in the art.

Abbreviation: DHP: 3,4-dihydro-2H-pyran; DIPEA: N,N-diisopropylethylamine; DMSO: dimethylsulfoxide; DSC: differential scanning calorimetry; DVS: dynamic vapor adsorption instrument; EtOAc or EA: ethyl acetate; h: hour; min: minutes; MsOH: methanesulfonic acid; NMP: N-methylpyrrolidone; XRPD: X-ray powder diffraction; 1H-NMR: hydrogen nuclear magnetic resonance spectrum; TGA: thermogravimetric analysis; PK: Pharmacokinetics.

Example 1 Preparation method of amorphous compound of formula I

Step 1: Preparation of compound S2

Add 801.11g of compound S1, 2492.50g of potassium acetate, 80g of purified water and 8L of acetic acid into the 20 L reaction kettle, stir and raise the temperature to 110±5°C, and react for 3 hours. HPLC detects that the reaction is complete.

Cool the system to 50°C, add the reaction solution to purified water (36.00 L) while stirring, stir for 30 minutes, filter, and rinse the filter cake with purified water (1.00 L). Transfer the filter cake to the reaction kettle, add purified water (10.00 L) and beat at room temperature for 0.5 hours, filter, and rinse the filter cake with purified water (2.00 L). The filter cake was naturally dried at room temperature for 48 hours to obtain S2 crude product. The crude product was beaten with ethyl acetate (2.00 L) for 1 hour, filtered, and the filter cake was rinsed with ethyl acetate (0.30 L). The filter cake was naturally dried at room temperature for 4 hours to obtain 701.35 g of S2 solid, yield: 92.91%.

Step 2: Preparation of compound S3

Add 701.23g S2 and 11.20L 1,4-dioxane to the 50L reaction kettle, stir and cool down; add 1185.75g potassium peroxybisulfate composite salt to 2.80 L water, stir for 5 minutes, and set aside. The temperature of the reaction system is lowered to 12°C, and the potassium hydrogen peroxysulfate compound salt mixture is added in batches, and the program controls the temperature to not exceed 20°C. After the addition is completed, the temperature is maintained at 10-20°C for 4 hours, and HPLC detects the reaction until it is completed.

Stop the reaction, add purified water (7.50 L) and methylene chloride (7.50 L) to the reaction kettle, stir for 10 minutes, and then separate into layers. The liquids were separated, and the aqueous phase was extracted once with dichloromethane (4.00 L). Combine the organic phases, wash with 0.5M sodium thiosulfate aqueous solution (1.00 L), purified water (7.00 L), purified water (7.00 L), saturated sodium chloride aqueous solution (7.00 L), and then wash with anhydrous sodium sulfate and anhydrous Dry over magnesium sulfate. Filter, and the filtrate is concentrated to obtain the crude product of S3. The crude product was beaten with tetrahydrofuran (2.20 L) for 18 hours, filtered, and the filter cake was rinsed with tetrahydrofuran (1.00 L). The filter cake was dried under vacuum at 25°C for 3 hours to obtain 472.31g of S3 solid, yield: 63.93%.

Step 3: Preparation of compound S21

Add 4.72L tetrahydrofuran, 472.31g S3, 252.69g DHP and 25.86g p-toluenesulfonic acid in sequence to the 10 L reaction kettle, stir and raise the temperature to 70°C, and keep the reaction for 3 hours. HPLC detects until the reaction is completed.

The system was cooled to 32°C and concentrated to dryness. Then add dichloromethane (4.50 L) and stir to dissolve. The organic phase was washed successively with purified water (3.00 L) and saturated aqueous sodium chloride solution (4.00 L), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated to dryness to obtain crude S21.

Step 4: Preparation of compound S22

While stirring, add 6L acetonitrile, 598.52g S21, 552.38g S04 and 129.24g DIPEA to the 20 L reaction kettle in sequence, then raise the temperature to 80°C, maintain the temperature for 21 hours, and detect by HPLC until the reaction is completed.

Cool to 60°C and concentrate. Approximately 3.00 L of acetonitrile was distilled off, and then 3.00 L of NMP was added. While stirring, pour the mixture into purified water (28.00 L). Stir for 20 minutes and filter. The filter cake was rinsed with purified water (1.00 L). Transfer the filter cake to the reaction kettle, add purified water (8.00 L) and beat at room temperature for 0.5 hours, filter, and rinse the filter cake with purified water (2.00 L). The filter cake was naturally dried at room temperature for 72 hours to obtain S22 crude product. Add tetrahydrofuran (2.00 L) to the crude product and beat for 18 hours, filter, and rinse the filter cake with n-heptane (1.00 L). The filter cake was naturally dried at room temperature to obtain 726.40 g of S22 solid.

Step 5: Preparation of amorphous compound of formula I

Under stirring, add 724.31g S22 and 5.79L methylene chloride to the 20 L reaction kettle. After the system is dissolved, filter and transfer the filtrate back to the 20 L reaction kettle. Stir and add 1.45L methanol. Add hydrogen chloride-1,4-dioxane solution (1.13L) dropwise, and control the temperature to not exceed 25°C. The reaction was stirred for 2 hours, and HPLC detected until the reaction was completed.

Concentrate to dryness, then add methanol (0.70 L) to dissolve all solids. While stirring, add the above mixture dropwise into dichloromethane (36.00 L). After the addition is complete, stir for 20 minutes. Filter, and rinse the filter cake with dichloromethane (2.00 L). The filter cake was naturally dried at room temperature for 15 hours, and 570.39 g of solid was obtained.

Add methanol (5.70 L) to the 50 L reaction kettle, then add the above solid (570.39 g), stir to dissolve, and filter. Transfer the filtrate back to the 20 L reaction kettle; stir and add purified water (1.00 L). Add 1M sodium bicarbonate aqueous solution (5.56 L) in batches, then add purified water (7.00 L) to dilute, and stir for 30 minutes. Filter and rinse with purified water (1.00 L). Transfer the filter cake to the reaction kettle, add purified water (7.00 L) and beat at room temperature for 20 minutes, filter, and rinse with purified water (2.00 L). The filter cake was naturally dried at room temperature for 48 hours to obtain 426.22g of a solid compound of formula I. According to XRPD measurement, the final product is amorphous.

Example 2-1 Preparation of mesylate crystal form A of the compound of formula I

Add 300 mL of tetrahydrofuran/water (19:1, v/v) mixed solvent to the 500 mL reaction bottle, then add 10 g of the amorphous compound of formula I, raise the temperature to 50 °C, stir and dissolve. Add 2.33 g methanesulfonic acid, stir at 50°C overnight, filter to obtain a solid, and dry it under vacuum at 40°C for 4 hours. The final product is measured by XRPD, TGA, DSC and 1H-NMR. The final product is the mesylate salt form of the compound of formula I. A. Use the same method to prepare more methanesulfonate crystal form A for later use.

Example 2-2 Preparation of methanesulfonic acid crystal form D of the compound of formula I

Weigh 1.00 g of the amorphous compound of formula I and 242 mg of methanesulfonic acid, add it to 10 mL of ethanol/water (19:1, v/v) mixed solvent, place it at 40°C for 1 hour, and stir at room temperature. After 4 hours, the solid was obtained by filtration and dried under vacuum at 40°C for 4 hours. After XRPD, TGA, DSC, 1H-NMR and single crystal diffraction measurements, the final product was the mesylate crystal form D of the compound of formula I. Use the same method to prepare more methanesulfonate crystal form D for later use.

Example 2-3 Preparation of mesylate crystal form A of the compound of formula I

Add tetrahydrofuran (4.47 L) and water (0.24 L) to the 10 L reaction kettle, start stirring, and raise the temperature to 50°C. 375.50 g of the amorphous compound of formula I are then added. Stir to dissolve, filter while hot, and transfer the filtrate to a 10 L reaction kettle. Weigh 16.04 g of methanesulfonic acid, dissolve it in tetrahydrofuran (0.94 L), and add it dropwise to the reaction solution. The addition will be completed in about 30 minutes.

Weigh 7.51 g of methanesulfonate crystal form D and add it. White turbidity precipitates. Keep the reaction solution at 50°C. Then dissolve 72.10 g of methanesulfonic acid in tetrahydrofuran (3.76 L) and add it dropwise to the reaction solution. The addition will be completed in about 3 hours. Cool to about 20°C and react for 2 hours, then filter. The filter cake was rinsed with tetrahydrofuran (0.40 L). The filter cake was vacuum dried at 50°C for 14 hours to obtain 353.24 g of methanesulfonate crystal form D.

Add ethyl acetate (5.70 L) to the 10 L reaction kettle, stir, and raise the temperature to 50°C. Then add the above sample (350.52 g), suspend and stir, then add 0.35 L of purified water, stir evenly, and then add 7.10 g of methanesulfonate crystal form A. Keep the reaction at 50°C. XRPD detects that the solid is completely transformed into methanesulfonate crystal form A. Cool to 20°C, mature for 1 hour and then filter with suction. The filter cake is rinsed with ethyl acetate (0.35 L). The filter cake is vacuumed at 50°C. Dry for 14 hours to obtain 352.16 g of solid. According to XRPD, TGA, DSC and 1H-NMR, the final product is the mesylate crystal form A of the compound of formula I.

Example 3 Preparation of benzene sulfonate crystal form A of the compound of formula I

Weigh about 300 mg of the amorphous compound of formula I and 230 mg of benzenesulfonic acid, add 6 mL of ethanol/water (19:1, v/v) solvent, place it at 50°C, stir magnetically overnight, and then filter to obtain The solid was vacuum dried at 40°C for 2 hours, and then dried at room temperature overnight. After XRPD, TGA, DSC and 1H-NMR measurements, the final product was benzene sulfonate crystal form A.

Example 4 Preparation of p-toluenesulfonate crystal form A of the compound of formula I

Weigh about 200 mg of the amorphous compound of formula I and 168.13 mg of p-toluenesulfonic acid, add 3 mL of ethanol solvent, place at room temperature, stir magnetically overnight, then raise the temperature to 35°C and stir overnight, centrifuge to obtain a solid, and incubate at 50 After vacuum drying for 4 hours at ℃, the final product was determined by XRPD, TGA, DSC and 1H-NMR to be p-toluenesulfonate crystal form A.

Example 5 Preparation of L-lactate crystal form A of the compound of formula I

Weigh about 300 mg of the amorphous compound of formula I and 123 mg of L-lactic acid, add 6 mL of ethanol solvent, stir at 50°C overnight, then cool at 0°C, stir at room temperature to precipitate a solid, and centrifuge. The solid was separated and dried under vacuum at 40°C for 3 hours. It was determined by XRPD, TGA, DSC and 1H-NMR that the final product was L-lactate crystal form A.

Example 6 Preparation of L-tartrate crystal form A of compound of formula I

Weigh about 200 mg of the amorphous compound of formula I and 144.37 mg of L-tartaric acid, add 4 mL of ethanol solvent, place at room temperature, stir magnetically overnight, then raise the temperature to 35°C and stir overnight, centrifuge to obtain a solid, and incubate at 50 After vacuum drying at ℃ for 4 hours, the final product was determined by XRPD, TGA, DSC and 1H-NMR to be L-tartrate crystal form A.

Example 7 Preparation of fumarate crystal form A of the compound of formula I

Weigh about 200 mg of the amorphous compound of formula I and 109.41 mg of fumaric acid, add 4 mL of ethyl acetate solvent, place at room temperature, stir magnetically overnight, then raise the temperature to 35°C and stir overnight, centrifuge to obtain a solid, and After vacuum drying at 50°C for 4 hours, the final product was determined by XRPD, TGA, DSC and 1H-NMR to be fumarate crystal form A.

Example 8 Preparation of L-malate crystal form A of compound of formula I

Weigh about 200 mg of the amorphous compound of formula I and 126.30 mg of L-tartaric acid, add 3 mL of ethyl acetate solvent, place at room temperature, stir magnetically overnight, then raise the temperature to 35°C and stir overnight, centrifuge to obtain a solid, and After vacuum drying at 50°C for 4 hours, the final product was determined by XRPD, TGA, DSC and 1H-NMR to be L-malate crystal form A.

Example 9 Preparation of Form A of Hydrochloride of Compound of Formula I

Weigh about 100 mg of the amorphous compound of formula I, add 2 mL of dilute hydrochloric acid (87 mg of concentrated HCl solution dissolved in 4 mL of H2O), suspend and stir at room temperature for 24 hours, then centrifuge, dry the solid under vacuum at room temperature for 4 hours, and According to XRPD, TGA and DSC measurements, the final product is hydrochloride crystal form A.

Example 10 Preparation of hydrochloride form B of compound of formula I

Weigh about 20 mg of the amorphous compound of formula I, add 0.5 mL of dilute hydrochloric acid (87 mg of concentrated HCl solution dissolved in 5 mL of acetone/water (19:1, volume ratio)), suspend at room temperature, stir for 48 hours, and then centrifuge. , the solid was vacuum dried at room temperature for 4 hours, and measured by XRPD, TGA and DSC, the final product was hydrochloride crystal form B.

Example 11 Preparation of hydrochloride form D of compound of formula I

Weigh about 20 mg of the amorphous compound of formula I, add 0.5 mL of dilute hydrochloric acid (87 mg of concentrated HCl solution dissolved in 5 mL of THF/H2O (19:1, v/v)), suspend at room temperature, stir for 48 hours, and then centrifuge. Separate, dry the solid under vacuum at room temperature for 4 hours, and determine by XRPD, TGA and DSC that the final product is hydrochloride crystal form D.

Example 12 Preparation of hydrochloride form E of the compound of formula I

Weigh about 100 mg of the amorphous compound of formula I, add 2 mL of dilute hydrochloric acid (182 mg of concentrated HCl solution dissolved in 4 mL of EtOH/H2O (19:1, v/v)), suspend at room temperature, stir for 2 days, and then centrifuge. Separate, dry the solid under vacuum at room temperature for 4 hours, and determine by XRPD and TGA that the final product is hydrochloride crystal form E.

Example 13 Preparation of hydrochloride form F of compound of formula I

Weigh about 100 mg of the amorphous compound of formula I, add 2 mL of hydrochloric acid diluent (87 mg of concentrated HCl solution dissolved in 4 mL of THF/H2O (19:1, v/v)), suspend at room temperature, stir for 24 hours, and then centrifuge. , determined by XRPD, the final product is hydrochloride crystal form F.

Example 14 Preparation of hydrochloride form G of compound of formula I

Weigh about 50 mg of the amorphous compound of formula I, add 1 mL of hydrochloric acid diluent (163 mg of concentrated HCl solution dissolved in 5 mL of THF/H2O (19:1, v/v)), suspend at room temperature, stir for 24 hours, and then centrifuge. Separate, dry the solid under vacuum at room temperature for 4 hours, and determine by XRPD, TGA and DSC that the final product is hydrochloride crystal form G.

Example 15 Preparation of hydrochloride form H of the compound of formula I

Weigh about 25 mg of the amorphous compound of formula I, add 0.5 mL of MEK, and then add 50 µL of 4M hydrochloric acid methanol solution, suspend and stir at room temperature for 48 hours, then centrifuge, dry the solid under vacuum at room temperature for 4 hours, and analyze by XRPD, TGA and According to DSC measurement, the final product is hydrochloride crystal form H.

Example 16 Preparation of Form I Hydrochloride of Compound of Formula I

Weigh about 20 mg hydrochloride crystal form B sample, add 0.4 mL acetonitrile/water (19:1, v/v) mixed solution, suspend and stir at room temperature for 5 days, then centrifuge, dry the solid in vacuum at room temperature for 4 hours, and According to XRPD, TGA and DSC measurements, the final product is hydrochloride crystal form I.

Example 17 Preparation of hydrochloride form J of the compound of formula I

Weigh about 20 mg hydrochloride crystal form B sample, add 0.4 mL water, suspend and stir at room temperature for 5 days, then centrifuge, dry the solid at room temperature under vacuum for 4 hours, and determine by XRPD, TGA and DSC that the final product is hydrochloride. Form J.

Example 18 Preparation of hydrochloride form K of compound of formula I

Weigh about 25 mg of the amorphous compound of formula I, add 0.5 mL of IPA, and then add 50 µL of 4M hydrochloric acid methanol solution, suspend and stir at room temperature for 48 hours, then centrifuge, dry the solid under vacuum at room temperature for 4 hours, and analyze by XRPD, TGA and According to DSC measurement, the final product is hydrochloride crystal form K.

Example 19 Preparation of hydrochloride form L of compound of formula I

Weigh about 100 mg of hydrochloride crystal form E sample and place it in an open air drying oven at 60°C for 10 days. After XRPD and TGA measurements, the final product is hydrochloride crystal form L.

Example 20 Preparation of hydrochloride form M of compound of formula I

Weigh about 15 mg hydrochloride crystal form G sample, dissolve it in 0.2 mL methanol, and slowly add it dropwise into 0.8 mL ethyl acetate. The turbid liquid obtained is centrifuged, and the solid is vacuum dried at room temperature for 4 hours, and analyzed by XRPD, TGA and DSC. It was determined that the final product was hydrochloride crystal form M.

Example 21 Preparation of hydrochloride form N of compound of formula I

Weigh about 25 mg hydrochloride crystal form E sample, add 0.4 mL MIBK solution, suspend and stir at 50°C for 7 days, then centrifuge, dry the solid at room temperature under vacuum for 3 hours, and determine by XRPD, TGA and DSC that the final product is hydrochloric acid. Salt crystal form N.

Example 22 Preparation of hydrochloride form O of compound of formula I

Weigh about 25 mg hydrochloride crystal form E sample, add 0.4 mL isopropyl alcohol solution, suspend and stir at 50°C for 7 days, and then centrifuge. The solid is vacuum dried at room temperature for 3 hours. After XRPD, TGA and DSC measurements, the final product is Hydrochloride salt form O.

Example 23 Preparation of hydrochloride form P of compound of formula I

Weigh about 25 mg hydrochloride crystal form E sample, add 0.4 mL cyclohexane solution, suspend and stir at 50°C for 7 days, and then centrifuge. The solid is vacuum dried at room temperature for 3 hours. After XRPD, TGA and DSC measurements, the final product is Hydrochloride salt form P.

Example 24 Preparation of hydrochloride form Q of compound of formula I

Weigh about 25 mg of the hydrochloride crystal form E sample, add 0.4 mL of methanol solution, suspend and stir at 50°C for 7 days, and then centrifuge. The solid is vacuum dried at room temperature for 3 hours. After XRPD, TGA and DSC measurements, the final product is hydrochloric acid. Salt crystal form Q.

Example 25 Preparation of hydrochloride form R of compound of formula I

Weigh about 500 mg of hydrochloride crystal form D sample, add 10 mL of ethyl acetate, suspend and stir at 60°C overnight and then filter with suction. The solid is vacuum dried at 45°C for 2 hours. After XRPD measurement, the final product is hydrochloride crystal form R. .

Example 26 Preparation of Form A of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of methanol, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD, TGA and DSC determination, the final product is crystalline form A of the compound of formula I.

Example 27 Preparation of Form B of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of ethanol, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD, TGA and DSC determination, the final product is crystal form B of the compound of formula I.

Example 28 Preparation of Form C of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of acetone, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD, TGA and DSC determination, the final product is crystal form C of the compound of formula I.

Example 29 Preparation of Form D of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of butanone, stir at room temperature for 4 hours, then centrifuge, dry, and determine by XRPD and TGA that the final product is crystal form D of the compound of formula I.

Example 30 Preparation of Form E of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of ethyl acetate, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD, TGA and DSC determination, the final product is crystal form E of the compound of formula I.

Example 31 Preparation of Form F of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of tetrahydrofuran, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD, TGA and DSC determination, the final product is crystal form F of the compound of formula I.

Example 32 Preparation of Form G of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of acetonitrile, stir at room temperature for 4 hours, and then centrifuge and dry. After XRPD and TGA determination, the final product is crystal form G of the compound of formula I.

Example 33 Preparation of Form H of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of acetonitrile/water (19:1, v/v), stir at room temperature for 4 hours, then centrifuge, dry, and determine the final product by XRPD, TGA and DSC. It is the crystal form H of the compound of formula I.

Example 34 Preparation of Form I of Compound of Formula I

Weigh about 15 mg of the amorphous compound of formula I, add it to 0.3 mL of ethanol/water (19:1, v/v), stir at room temperature for 4 hours, then centrifuge, dry, and measure by XRPD and TGA. The final product is of formula Compound I, Form I.

Example 35 Preparation of Form J of Compound of Formula I

About 15 mg of the amorphous compound of formula I was weighed, heated to 220°C, and measured by XRPD, TGA and DSC. The final product was crystal form J of compound of formula I.

Example 36 Ion chromatography measurement

Take about 20 mg of methanesulfonate crystal form A to prepare a solution with a concentration of 0.1 mg/mL. Using no sodium mesylate as the reference substance, measure the methanesulfonic acid content in methanesulfonate crystal form A by ion chromatography. The results are shown in Table 6.

Table 6 Crystal form Methanesulfonate molecular weight Free molecular weight Theoretical content of methanesulfonic acid in monomethanesulfonate Actual measured content of methanesulfonic acid in mesylate crystal form A Salt formation ratio of mesylate crystal form A Mesylate crystal form A 548.66 452.55 17.52% 18.40% 1:1.05

The results show that the ratio of the theoretical content of methanesulfonic acid in methanesulfonate to the actual measured content of methanesulfonic acid in mesylate crystal form A is 1:1.05, proving that methanesulfonate crystal form A is monomethanesulfonate. Acid.

Example 37 Single crystal diffraction experiment

Measured according to the first method of Part Four General Chapter 0451 of the 2020 edition of the "Chinese Pharmacopoeia", test conditions: CuKα radiation, φ/ω scanning. Single crystal diffraction instrument and equipment information is shown in Table 8.

Single crystal diffraction experiment results: The single crystal data of methanesulfonate crystal form D is shown in Table 9. The three-dimensional structure diagram of its asymmetric unit is shown in Figure 5, and its unit cell stacking diagram is shown in Figure 5-1.

Table 7 Single crystal diffraction instrument and equipment information instrument Model production unit Instrument number Single crystal X-ray diffractometer Bruker SMART APEX-II Bruker AXS GmbH, Germany IARC-032-SXRD-01

Table 8 Single crystal data of mesylate form D of compound of formula I Molecular formula C28H32N6O4S molecular weight 548.66 Crystal system triclinic system space group P1 Unit cell parameters a=8.6411(1)Å, b=10.8194(2)Å, c=15.8226(2)Å; α=82.816(1), β=78.424(1), γ=71.392(1)° unit cell volume V=1370.46(3)A3 Number of asymmetric units in the unit cell (Z) 1 Calculate density 1.330g/cm3

Example 38 Transformation relationship of mesylate crystal form A/D of compound of formula I

Weigh a certain amount of methanesulfonate crystal form A sample, add tetrahydrofuran and acetonitrile respectively, suspend and stir overnight, and filter to obtain a saturated solution. Add 5 mg of crystal form A and 5 mg of crystal form D samples to the saturated solution respectively, stir at 5°C/25°C/60°C, and take samples for XRPD testing. The results are shown in Table 9.

Table 9 No. temperature Solvent Crystal form results 1 5℃ Tetrahydrofuran Mesylate crystal form A 2 Acetonitrile Mesylate crystal form A 3 25℃ Tetrafuran Mesylate crystal form A 4 Acetonitrile Mesylate crystal form A 5 60℃ Tetrahydrofuran Mesylate crystal form A 6 Acetonitrile Mesylate crystal form A

The results show that between 5℃ and 60℃, mesylate crystal form A is thermodynamically more stable than mesylate crystal form D.

Example 39 Purity Determination

Take the crystalline compounds prepared by the methods of Examples 2-1, 2-2, and Examples 3 to 8, and measure the purity by HPLC. The result is as follows: Compound crystal form test article Purity (%) Mesylate crystal form A Example 2-1 99.58 Mesylate crystal form D Example 2-2 99.92 Benzene sulfonate crystal form A Example 3 97.96 p-Toluenesulfonate Crystal Form A Example 4 97.84 L-Lactate Form A Example 5 98.66 L-tartrate crystal form A Example 6 98.44 Fumarate crystal form A Example 7 93.21 L-malate crystal form A Example 8 97.64

Surprisingly, compared with other crystal forms, mesylate crystal form A and mesylate crystal form D have achieved unexpected purification effects, that is, they can be produced with higher purity. It can be seen that mesylate crystal form A and mesylate crystal form D are suitable for preparing more stable and high-quality drugs.

Example 40 Determination of stability

The stability of the crystalline forms of the invention was determined. Combine mesylate crystal form A, mesylate crystal form D, benzenesulfonate crystal form A, p-toluenesulfonate crystal form A, L-lactate crystal form A, L-tartrate crystal form A, Fumarate crystal form A and L-malate crystal form A were respectively placed in a 4500 Lux light environment, a 60°C high temperature environment and a 90% RH ± 5% RH high humidity environment for stability testing. On the 0th day, the Samples were taken on the 5th, 10th and 30th days. The appearance properties, moisture content, and total related substances of the samples were recorded and compared with the initial data. The comparison results show that mesylate crystal form A, mesylate crystal form D, benzenesulfonate crystal form A, p-toluenesulfonate crystal form A, L-lactate crystal form A, and L-tartrate crystal Form A, fumarate crystal form A and L-malate crystal form A have better stability, especially methanesulfonate crystal form A and methanesulfonate crystal form D have better stability.

Example 41 Determination of solubility

Weigh approximately 10 mg of the mesylate crystal form A, benzenesulfonate crystal form A, p-toluenesulfonate crystal form A, L-tartrate crystal form A, and fumarate crystal form A of the compound of formula I respectively. , L-malate crystal form A and hydrochloride crystal form N, add 0.8 mL of artificial gastric juice or purified water, place the suspension in a constant temperature shaker at 37 °C and shake for 24 h, and take the mother liquor to detect the concentration. The results are shown in Table 10.

Table 10 Solubility measurement results of different crystal forms Compound crystal form Solubility at 37°C (mg/mL) pure water artificial gastric juice Mesylate crystal form A 2.8 1.5 Benzene sulfonate crystal form A 0.2 1.0 p-Toluenesulfonate Crystal Form A 0.03 0.2 L-tartrate crystal form A 2.3 0.9 Fumarate crystal form A 2.4 1.5 L-malate crystal form A 3.3 2.6 Hydrochloride crystal form N 3.7 2.1 Amorphous form of compound of formula I 0.005 N/A

Example 42 Determination of dynamic moisture adsorption (DVS)

The dynamic adsorption instrument detection equipment and method of the present invention are shown in Table 11, and the dynamic moisture adsorption measurement results are shown in Table 12.

Table 11 Dynamic moisture adsorption instrument testing equipment and method data Device name Dynamic vapor adsorption instrument Manufacturer Surface Measurement Systems Device model DVS Resolution Sample tray Aluminum crucible Test sample size 30-50 mg protective gas Nitrogen Gas flow rate 200sccm

Table 12 Dynamic moisture adsorption measurement results Compound crystal form Weight change in the range of 0%RH to 80%RH Mesylate crystal form A 1.30% Mesylate crystal form D 0.11% p-Toluenesulfonate Crystal Form A 2.50% L-Lactate Form A 3.10% L-tartrate crystal form A 6.40% Fumarate crystal form A 1.80% L-malic acid crystal form A 3.80% Hydrochloride crystal form N 2.10%

It can be seen from the test results that the weight change of methanesulfonate crystal form D is 0.11% in the range of 0%RH to 80%RH. It has no or almost no hygroscopicity and has outstanding advantages in hygroscopicity; and compared with In terms of hygroscopicity data of other crystal forms, mesylate crystal form A also has obvious advantages.

Example 43 In vivo pharmacokinetics (PK) comparative experiment

Experimental method: 15 male rats were used and divided into five groups, with 3 rats in each group. Each group was administered 10 mg/kg mesylate crystal form A and p-toluenesulfonate crystal form of the compound of formula (I) in a single administration. A, L-lactate crystal form A, L-tartrate crystal form A, L-malate crystal form A and hydrochloride crystal form J, respectively at specified time points (15min, 30min, 1h, 2h, 4h , 7h and 24h) blood was collected from the fundus venous plexus, the plasma was separated, and stored in a -80°C refrigerator.

Take 30 or 50 μL of plasma and add it to 200 μL of acetonitrile containing the internal standard to precipitate, centrifuge, take 100 μL of the supernatant, add 100 μL of water and mix well, take 10 μL to LC-MS/MS for detection, the test data are shown in Table 13 :

Table 13 Comparative experimental results of pharmacokinetics in vivo compound Dosage (mg/kg) Cmax (ng/mL) AUClast（h*ng/mL） Mesylate crystal form A 10 1177 5489 p-Toluenesulfonate Crystal Form A 10 1255 5613 L-Lactate Form A 10 1613 7916 L-tartrate crystal form A 10 979 5328 L-malate crystal form A 10 1887 9387 Hydrochloride crystal form J 10 2277 9317

It was found that the above crystalline form has high bioavailability in the body and meets the needs of drug development.

Pharmacological experiments

Example A: Assay of SHP2 allosteric inhibitory enzyme activity

SHP2 is allosterically activated by binding of a bis-tyrosinyl-phosphopeptide to its Src homology 2 (SH2) domain. This subsequent activation step results in the release of the autoinhibitory interface of SHP2, which in turn makes the SHP2 protein tyrosine phosphatase (PTP) activated and available for substrate recognition and reaction catalysis. The catalytic activity of SHP2 was monitored using the surrogate DiFMUP in a rapid fluorescence assay format.

Test steps:

(1) Compound preparation:

Use 100% DMSO to dilute the compound of the present invention (10mM stock solution) to an appropriate multiple. The final test concentrations of the compound of the present invention are 1 μM, 0.333 μM, 0.111 μM, 0.0370 μM, 0.0124 μM, 0.00412 μM, 0.00137 μM, 0.00046 μM, 0.00015 μM, 0.00 μM;

(2) Prepare enzyme reaction working solution:

SHP2 enzyme activity assay was performed in a 96-well black polystyrene plate (flat bottom, low flange, non-binding surface) (Perki Elmer, Cat#6005270) at room temperature using a final reaction volume of 50 μL and the following assay buffer conditions. : 60mM HEPES, 75mM NaCl, 75mM KCl, 0.05% BRIJ-35, 1mM EDTA, 5mM DTT.

(3) Enzyme catalyzed reaction and data monitoring:

Take the compound of the present invention and add it to the corresponding 96-well plate, and set the blank test well without adding compound and enzyme and only adding buffer. Melt SHP2 Activating Peptide (IRS1_pY1172(dPEG8)pY1222) on ice, add 0.5 μM to each well, then add 0.2 ng SHP2 protein sample to the corresponding well plate, and incubate at room temperature for 1 hour. Add the substrate DiFMUP (Invitrogen, Cat#D6567) and react at room temperature for 1 hour. Fluorescent signals were monitored using a microplate reader (Envision, Perki Elmer) using excitation and emission wavelengths of 340 nM and 450 nM, respectively.

(4) Data analysis:

Calculation formula:

Inhibition rate %=[1-( Conversion_ _sample -Conversion_ _min )/( Conversion_ _max -Conversion_ _min )]×100%

Among them: Conversion_sample is the conversion rate reading of the sample; Conversion_min is the average value of the blank control wells, which represents the conversion rate reading of the wells without enzyme activity; Conversion_max is the average ratio of the positive control wells, which represents the conversion rate reading of the wells without compound inhibition.

The log(inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism was used to fit the dose-effect curve, and the IC50 value of the compound on the enzyme activity was calculated. It has been determined that the IC50 of the compound of formula I of the present invention against SHP2 is 0.9 nM.

Example B: Cell Proliferation Assay

In vitro cell assays were used to evaluate the effects of the compounds of the present invention on the proliferation of leukemia cells MV-4-11 and lung cancer cells NCI-H358. The detection method used in the experiment is the CELL TITER-GLO (CTG) luminescence method, which can detect the number of viable cells by quantitatively measuring ATP. Because ATP participates in a variety of enzymatic reactions in organisms and is an indicator of living cell metabolism, its content directly reflects the number and status of cells. During the experiment, CellTiter-Glo™ reagent was added to the cell culture medium and the luminescence value was measured. The amount of ATP is directly proportional, and ATP is positively related to the number of viable cells. Therefore, cell viability can be examined by detecting ATP content.

Test steps:

(1) Cell plating:

Take a bottle of MV-4-11 cells in the logarithmic growth phase, centrifuge and resuspend the cells and count them. Adjust the cell density and inoculate them into a 96-well plate (Corning #3917). Inoculate 4000 cells in each well. Place the well plate at 37°C. After culturing for 24 hours in an incubator with 5% CO2, add the compound of the present invention for treatment;

Take a bottle of NCI-H358 cells in the logarithmic growth phase, digest and resuspend the cells, count them, adjust the cell density and inoculate them into a 96-well transparent ultra-low adsorption cell culture plate (Corning #3474). Inoculate 2000 cells in each well of the plate. Place it in an incubator at 37°C and 5% CO2 for 24 hours and then add the compound of the present invention for treatment;

(2) Cell compound processing:

Prepare an appropriate amount of the compound of the present invention for cell treatment. The final concentration of the compound from high to low is 1000 nM, 333.3 nM, 111.1 nM, 37.04 nM, 12.35 nM, 4.115 nM, 1.372 nM, 0.4572 nM, 0.1524 nM, 0 nM. Place the plate into a 37°C, 5% _CO2 incubator for 120 hours. The wells that only added culture medium without adding cells were set as the blank group; the group with a compound concentration of 0 nM was set as the zeroing group.

(3) CTG detection:

After NCI-H358 cells were cultured for 96 hours, add 50 μL of CellTiter-Glo® Luminescent Cell Viability Assay solution to each well, shake gently for 2 minutes, and continue to incubate at room temperature for 10 minutes. The cell reaction system was transferred to a white-bottomed 96-well plate. Read the detection value of each well on a multifunctional microplate reader.

After culturing MV-4-11 cells for 120 hrs, add 50 μL of CellTiter-Glo® Luminescent Cell Viability Assay solution to each well, shake gently for 2 minutes, continue to incubate at room temperature for 10 minutes, and read each well on a multifunctional microplate reader. detection value.

(4) Data analysis:

Calculate the inhibition rate based on the luminescence value reading, Inhibition rate % = (1 - (administration group value - blank group value)/(zero adjustment group value - blank group value) × 100

GraphPad Prism's log(inhibitor) vs. response-Variable slope fits the dose-response curve and calculates the IC50 of the compound that inhibits cell proliferation. The IC50 value of the compound of Chinese formula I of the present invention on MV-4-11 cells and the IC50 value on NCI-H358 cells are both less than 20 nM.

Example C Patch-clamp experiment to evaluate the effect of compounds on hERG ion channels

Test solution recipe:

Dilute the test sample diluent sequentially with extracellular fluid to prepare detection solutions with final concentrations of 0.3 µM, 1 µM, 3 µM, 10 µM and 30 µM. Visually inspect the solubility of the sample to be tested.

Cell culture and plating:

The cell line is derived from HEK293 cells and cultured in a 37°C, 5% CO ₂ incubator. In order to prevent cell senescence caused by contact inhibition, when the cell culture confluence should not exceed 80%, it should be passaged every 3/4 days, and the inoculation density of each T175 bottle should be 2*106 cells. First pre-wash with phosphate buffered saline (PBS), then digest the cells with trypsin/EDTA for 2-3 minutes, add cell culture medium to terminate digestion, and transfer to a new culture bottle.

HEK293 cells overexpressing hERG potassium ion channel were cultured overnight at a cell density lower than 50%. The experimental cells were transferred to a cell bath embedded in an inverted microscope platform (Diaphot, Nikon), and extracellular fluid was perfused. The extracellular fluid contained 137 mM NaCl, 4 mM KCl, 1.8 mM CaCl ₂ , 1 mM MgCl ₂ , 10 mM glucose and 10 mM HEPES (pH 7.4 with NaOH), and the perfusion rate was 4 ml/min. The inner tube solution contains 130 mM KCl, 1 mM MgCl ₂ , 5 mM EGTA, 5 mM MgATP and 10 mM HEPES (pH 7.2 with KOH). Membrane currents were recorded using a HEKA EPC-10 patch-clamp amplifier and a PATCHMASTER acquisition system (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany). All experiments were performed at room temperature (22 -23°C).

The electrode (BF150-86-10) was straightened using a P-97 microelectrode puller (Sutter Instrument Company, One DigitalDrive, Novato, CA 94949). The inner diameter of the electrode is 1-1.5 mm, and the water inlet resistance after being filled with internal fluid is 2-4 MΩ.

Electrophysiological stimulation program:

After forming a whole-cell seal, wait for the current to stabilize for 2 minutes (the current attenuation is less than 5% within 5 minutes, and the tail current value is greater than 500 pA). At this time, the peak value of the tail current is the control current value. Then the extracellular fluid containing the drug to be tested is perfused. The same procedure was repeated 3 to 5 times, exposing each cell to 4 to 6 compounds with increasing concentrations. The process of blocking and deactivating hERG during compound exposure and washout was continuously recorded.

When whole-cell sealing is formed, the cell membrane voltage is clamped at -80 mV, and depolarization is performed for 2 seconds every 12 seconds. The clamping voltage is depolarized from -80 mV to -50 mV, and under a 5-second repolarization pulse of -50 mV Measure the peak value of the tail current.

Parameter analysis:

Peak hERG tail currents were measured under a 5 sec repolarizing pulse at -50 mV. The current inhibition rate was plotted for each drug concentration as a function of the logarithm of the compound concentration. Use the following Hill equation to fit the concentration response curve and fit the IC50.

Y: test value; Bottom: the lowest test value (0); Top: the highest test value (1); Hillcoefficient: the maximum absolute value of the curve slope.

Data Analysis and Statistics

Experimental data were collected using PATCHMASTER (HEKA Instruments Inc., D-67466 Lambrecht, Pfalz, Germany) and analyzed and statistically analyzed using Origin (OriginLab Corporation, Northampton, MA).

Data are presented as mean ± standard deviation. Use t-Test test to determine whether there is a significant difference compared with the control group. When p < 0.05, it is considered to have a significant difference.

After measurement, the hERG IC50 value of the compound of formula I of the present invention is 9.5 μM.

Example D: In vitro metabolic stability of human and rat liver microsomes

Test buffer preparation:

Dissolve 1900 mg MgCl2 into 400 mL of ultrapure water.

Dissolve 17.42 g K2HPO4 and 13.65 g KH2PO4 into 1000 mL of ultrapure water respectively. Mix a certain proportion of K2HPO4 and KH2PO4 and adjust the pH to 7.30 ± 0.10.

Reaction stop solution preparation:

Prepare a solution containing 10 ng/mL labetalol and 10 ng/mL glyburide in acetonitrile and store at 4°C.

Preparation of compound working solution:

Use DMSO to prepare 10 mM verapamil (positive control) and test sample stock solutions. Then use MeOH/ACN/H2O solution (volume ratio 1:1:2) to dilute into 50 µM and 200 µM working solutions respectively.

Test steps:

(1) Add 40 μL MgCl2 and 306 μL PBS into 96 plate wells (set up a blank control group, a positive control group, a compound sample group to be tested, and a negative control group at the same time).

(2) Add 4 μL compound working solution to each well (for the blank group, add 4 μL PBS buffer) (Note: The final concentration of DMSO volume in the system is ≤0.5%).

(3) Add 10 μL liver microsomes (concentration: 20 mg/mL) to each well, mix well, and pre-incubate at 37°C for 10 minutes.

(4) Add 40 μL of 10 mM NADPH working solution to each well to start the reaction (an equal volume of PBS buffer is added to the blank control group and negative control group respectively), and the total reaction volume is 400 μL.

(5) Take out 50 μL samples from the above reaction solution at 0, 5, 15, and 45 minutes respectively, and add 400 μL reaction stop solution to terminate the reaction.

(6) Shake the sample after terminating the reaction on the oscillator for 5 minutes

(7) Centrifuge the sample at 3200 rcf for 10 minutes. After the centrifugation is completed, transfer 50 μL of the supernatant to 200 μL H2O and dilute it for LC-MS/MS analysis.

data analysis

Calculation of t1/2 and CLint using first-order kinetic equations: k=-slope t1/2 = 0.693/k CLint = k/Cprotein In the formula, k represents the elimination constant, calculated from the logarithmic linear plot of remaining percentage versus time. t1/2 represents half-life. Cprotein is the concentration of liver microsomes. The results of liver microsomal metabolism stability of compounds of formula I in humans and rats are shown in Table 14.

Table 14 Liver microsomal metabolic stability data of compounds of formula I in humans and rats sample CLint (µL/min/mg protein) people rat Compounds of formula I 10.6 17.4

without

Figure 1 is an X-ray powder diffraction pattern of the mesylate crystal form A of the compound of formula I; Figure 2 is a differential scanning calorimetry spectrum of the mesylate crystal form A of the compound of formula I; Figure 3 is a thermogravimetric analysis spectrum of the mesylate crystal form A of the compound of formula I; Figure 4 is a 1H-NMR spectrum of the mesylate crystal form A of the compound of formula I; Figure 5 is a three-dimensional structure diagram of the asymmetric unit of the mesylate crystal form D of the compound of formula I; Figure 5-1 is a unit cell stacking diagram of the mesylate crystal form D of the compound of formula I; Figure 6 is an X-ray powder diffraction pattern of the mesylate crystal form D of the compound of formula I; Figure 7 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the mesylate crystal form D of the compound of formula I; Figure 8 is a 1H-NMR spectrum of the mesylate crystal form D of the compound of formula I; Figure 9 is an X-ray powder diffraction pattern of the benzene sulfonate crystal form A of the compound of formula I; Figure 10 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the benzene sulfonate crystal form A of the compound of formula I; Figure 11 is a 1H-NMR spectrum of the benzene sulfonate crystal form A of the compound of formula I; Figure 12 is an X-ray powder diffraction pattern of the p-toluenesulfonate crystal form A of the compound of formula I; Figure 13 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the p-toluenesulfonate crystal form A of the compound of formula I; Figure 14 is a 1H-NMR spectrum of the p-toluenesulfonate crystal form A of the compound of formula I; Figure 15 is an X-ray powder diffraction pattern of the L-lactate crystal form A of the compound of formula I; Figure 16 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the L-lactate crystal form A of the compound of formula I; Figure 17 is a 1H-NMR spectrum of compound L-lactate crystal form A of formula I; Figure 18 is an X-ray powder diffraction pattern of compound L-tartrate crystal form A of formula I; Figure 19: is the thermogravimetric analysis spectrum of the L-tartrate crystal form A of the compound of formula I; Figure 20: is the 1H-NMR spectrum of compound L-tartrate crystal form A of formula I; Figure 21: is the X-ray powder diffraction pattern of the fumarate crystal form A of the compound of formula I; Figure 22: is the thermogravimetric analysis spectrum of the fumarate crystal form A of the compound of formula I; Figure 23: 1H-NMR spectrum of fumarate crystal form A of the compound of formula I; Figure 24: is the X-ray powder diffraction pattern of the L-malate crystal form A of the compound of formula I; Figure 25: is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the L-malate crystal form A of the compound of formula I; Figure 26: is the 1H-NMR spectrum of compound L malate crystal form A of formula I; Figure 27: is the X-ray powder diffraction pattern of the hydrochloride crystal form A of the compound of formula I; Figure 28: is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form A of the compound of formula I; Figure 29: is the X-ray powder diffraction pattern of the hydrochloride crystal form B of the compound of formula I; Figure 30: is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form B of the compound of formula I; Figure 31: is the X-ray powder diffraction pattern of the hydrochloride crystal form D of the compound of formula I; Figure 32: is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form D of the compound of formula I; Figure 33: is the X-ray powder diffraction pattern of the hydrochloride crystal form E of the compound of formula I; Figure 34: is the thermogravimetric analysis spectrum of the hydrochloride crystal form E of the compound of formula I; Figure 35: is the X-ray powder diffraction pattern of the hydrochloride crystal form F of the compound of formula I; Figure 36: is the X-ray powder diffraction pattern of the hydrochloride crystal form G of the compound of formula I; Figure 37: is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form G of the compound of formula I; Figure 38: is the X-ray powder diffraction pattern of the hydrochloride crystal form H of the compound of formula I; Figure 39 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form H of the compound of formula I; Figure 40 is an X-ray powder diffraction pattern of the hydrochloride crystal form I of the compound of formula I; Figure 41 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form I of the compound of formula I; Figure 42 is an X-ray powder diffraction pattern of the hydrochloride crystal form J of the compound of formula I; Figure 43 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form J of the compound of formula I; Figure 44 is an X-ray powder diffraction pattern of the hydrochloride crystal form K of the compound of formula I; Figure 45 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form K of the compound of formula I; Figure 46 is an X-ray powder diffraction pattern of the hydrochloride crystal form L of the compound of formula I; Figure 47 is a thermogravimetric analysis spectrum of the hydrochloride crystal form L of the compound of formula I; Figure 48 is an X-ray powder diffraction pattern of the hydrochloride crystal form M of the compound of formula I; Figure 49 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form M of the compound of formula I; Figure 50 is an X-ray powder diffraction pattern of the hydrochloride crystal form N of the compound of formula I; Figure 51 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form N of the compound of formula I; Figure 52 is an X-ray powder diffraction pattern of the hydrochloride crystal form O of the compound of formula I; Figure 53 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form O of the compound of formula I; Figure 54 is an X-ray powder diffraction pattern of the hydrochloride crystal form P of the compound of formula I; Figure 55 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form P of the compound of formula I; Figure 56 is an X-ray powder diffraction pattern of the hydrochloride crystal form Q of the compound of formula I; Figure 57 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the hydrochloride crystal form Q of the compound of formula I; Figure 58 is an X-ray powder diffraction pattern of the hydrochloride form R of the compound of formula I; Figure 59 is an X-ray powder diffraction pattern of the crystal form A of the compound of formula I; Figure 60 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the crystal form A of the compound of formula I; Figure 61 is an X-ray powder diffraction pattern of the crystal form B of the compound of formula I; Figure 62 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the compound of formula I, Form B; Figure 63 is an X-ray powder diffraction pattern of the crystal form C of the compound of formula I; Figure 64 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the crystal form C of the compound of formula I; Figure 65 is an X-ray powder diffraction pattern of the crystal form D of the compound of formula I; Figure 66 is a thermogravimetric analysis spectrum of the crystalline form D of the compound of formula I; Figure 67 is an X-ray powder diffraction pattern of the crystal form E of the compound of formula I; Figure 68 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the crystalline form E of the compound of formula I; Figure 69 is an X-ray powder diffraction pattern of the crystal form F of the compound of formula I; Figure 70 is an overlay of a differential scanning calorimetry spectrum and a thermogravimetric analysis spectrum of the compound of Formula I, Form F; Figure 71 is an X-ray powder diffraction pattern of the crystal form G of the compound of formula I; Figure 72 is a thermogravimetric analysis spectrum of the crystalline form G of the compound of formula I; Figure 73 is an X-ray powder diffraction pattern of the crystal form H of the compound of formula I; Figure 74 is an overlay of a differential scanning calorimetry spectrum and a thermogravimetric analysis spectrum of the compound of Formula I, Form H; Figure 75 is an X-ray powder diffraction pattern of the crystal form I of the compound of formula I; Figure 76 is a thermogravimetric analysis spectrum of the crystal form I of the compound of formula I; Figure 77 is an X-ray powder diffraction pattern of the crystal form J of the compound of formula I; Figure 78 is an overlay of the differential scanning calorimetry spectrum and the thermogravimetric analysis spectrum of the compound of formula I, Form J; Figure 79 is an X-ray powder diffraction pattern of the amorphous compound of formula I.

Unless otherwise stated, the detection instrument information and detection method parameters used in the present invention are as shown in Tables 2 to 5:

Table 2 Device name X-ray powder diffractometer (XRPD) Device model Bruker D8 Advance Technical indicators Kα radiation (40 kV, 40 mA) with copper target wavelength of 1.54Å, θ-2θ goniometer, Ni monochromator, Lynxeye detector Detection parameters Detection angle 3-40°2θ step size 0.02°2θ Scan time per step (s) 0.2 or 0.3 or 0.6

table 3 Device name Thermogravimetric Analyzer (TGA) Device model Discovery TGA 550 Sample tray Platinum crucible + aluminum crucible protective gas Nitrogen Gas flow rate (sample purge) 40mL/min Gas flow rate (balance) 60mL/min heating rate 10℃/min

Table 4 Device name Differential Scanning Calorimeter (DSC) Device model Discovery DSC 2500 Sample tray Aluminum crucible protective gas Nitrogen Gas flow rate 50mL/min heating rate 10℃/min

table 5 Device name NMR Manufacturer Brooke Device model AVANCE NEO 500 Deuterated reagent Deuterated DMSO Detection parameters NS: 16 D1: 1[sec] O1P: 6.175ppm SW: 19.9955ppm TE: 298K

Claims (64)

A salt form of the compound represented by formula I: Formula I. The salt form of the compound represented by formula I as described in claim 1, wherein the salt form is methanesulfonate, benzenesulfonate, p-toluenesulfonate, L-lactate, L-tartrate, rich Malate, L-malate and hydrochloride. The salt form of the compound represented by formula I as described in claim 1 or 2, wherein the salt form is a methanesulfonate salt. The salt form of the compound represented by Formula I according to any one of claims 1 to 3, wherein the salt form is a crystalline form. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is methanesulfonate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, and 23.5°±0.2°. The salt form of the compound represented by formula I according to any one of claims 1 to 5, wherein the salt form is methanesulfonate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, 16.9°±0.2°, and 23.5°±0.2°. The salt form of the compound represented by formula I according to any one of claims 1 to 6, wherein the salt form is mesylate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 5.6°±0.2°, 7.2°±0.2°, 10.0°±0.2°, 14.5°±0.2°, 16.9°±0.2°, 19.8°±0.2°, 20.6°±0.2°, 22.5°±0.2°, 23.5° Characteristic peak of ±0.2°. The salt form of the compound represented by formula I according to any one of claims 1 to 7, wherein the salt form is methanesulfonate crystal form A, and its X-ray powder diffraction pattern is basically as shown in Table 1 Show. The salt form of the compound represented by formula I according to any one of claims 1 to 8, wherein the salt form is methanesulfonate crystal form A, and its X-ray powder diffraction pattern is basically as shown in Figure 1 Show. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is methanesulfonate crystal form D, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 5.7°±0.2°, 16.4°±0.2°, 19.3°±0.2°, 20.1°±0.2°, and 22.9°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 10, wherein the salt form is methanesulfonate crystal form D, and its X-ray powder diffraction pattern has a diffraction angle 2θ is 5.7°±0.2°, 11.9°±0.2°, 16.4°±0.2°, 17.3°±0.2°, 19.3°±0.2°, 20.1°±0.2°, 22.9°±0.2°, 26.1°±0.2° Characteristic peaks. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 10 to 11, wherein the salt form is methanesulfonate crystal form D, and its X-ray powder diffraction pattern is basically As shown in Figure 6. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is benzene sulfonate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 8.4°±0.2°, 11.0°±0.2°, 14.2°±0.2°, 16.8°±0.2°, 17.4°±0.2°, 18.8°±0.2°, and 25.6°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 13, wherein the salt form is benzenesulfonate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ is 6.2°±0.2°, 8.4°±0.2°, 11.0°±0.2°, 11.8°±0.2°, 13.8°±0.2°, 14.2°±0.2°, 15.9°±0.2°, 16.8°±0.2°, The characteristic peaks are 17.4°±0.2°, 18.8°±0.2°, and 25.6°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 13 to 14, wherein the salt form is benzenesulfonate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 9. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is p-toluenesulfonate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ The characteristic peaks are 6.0°±0.2°, 10.4°±0.2°, 14.2°±0.2°, 18.7°±0.2°, and 22.3°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 16, wherein the salt form is p-toluenesulfonate crystal form A, and its X-ray powder diffraction pattern has diffraction Angle 2θ is 6.0°±0.2°, 10.4°±0.2°, 11.2°±0.2°, 12.4°±0.2°, 13.0°±0.2°, 14.2°±0.2°, 18.3°±0.2°, 18.7°±0.2° , a characteristic peak of 22.3°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 16 to 17, wherein the salt form is p-toluenesulfonate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 12 above. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is L-lactate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 6.2°±0.2°, 6.7°±0.2°, 12.2°±0.2°, 13.9°±0.2°, 18.5°±0.2°, 21.0°±0.2°, and 25.6°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 19, wherein the salt form is L-lactate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ is 6.2°±0.2°, 6.7°±0.2°, 12.2°±0.2°, 13.9°±0.2°, 17.2°±0.2°, 17.9°±0.2°, 18.5°±0.2°, 21.0°±0.2°, The characteristic peaks are 25.1°±0.2° and 25.6°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 19 to 20, wherein the salt form is L-lactate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 15. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is L-tartrate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 5.9°±0.2°, 12.6°±0.2°, 15.8°±0.2°, 18.9°±0.2°, and 24.6°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 22, wherein the salt form is L-tartrate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ is 5.9°±0.2°, 10.8°±0.2°, 12.6°±0.2°, 15.1°±0.2°, 15.8°±0.2°, 17.9°±0.2°, 18.6°±0.2°, 18.9°±0.2°, The characteristic peaks are 19.4°±0.2°, 24.6°±0.2°, and 25.3°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 22 to 23, wherein the salt form is L-tartrate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 18. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is fumarate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of The characteristic peaks are 7.5°±0.2°, 15.0°±0.2°, 16.2°±0.2°, 17.3°±0.2°, 22.8°±0.2°, and 28.7°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 25, wherein the salt form is fumarate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ is the characteristic peak of 7.5°±0.2°, 15.0°±0.2°, 16.2°±0.2°, 17.3°±0.2°, 22.8°±0.2°, 28.7°±0.2°, and 29.3°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 25 to 26, wherein the salt form is fumarate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 21. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is L-malate crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ They are the characteristic peaks of 5.1°±0.2°, 10.2°±0.2°, 17.2°±0.2°, 20.6°±0.2°, and 23.0°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 28, wherein the salt form is L-malate crystal form A, and its X-ray powder diffraction pattern has diffraction The angle 2θ is the characteristic peak of 5.1°±0.2°, 6.1°±0.2°, 10.2°±0.2°, 16.2°±0.2°, 17.2°±0.2°, 20.6°±0.2°, and 23.0°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 28 to 29, wherein the salt form is L-malate crystal form A, and its X-ray powder diffraction pattern is basically As shown in Figure 24 above. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is hydrochloride crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 6.9 The characteristic peaks are °±0.2°, 10.4°±0.2°, 16.2°±0.2°, 16.9°±0.2°, 18.1°±0.2°, 18.6°±0.2°, and 19.5°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 31, wherein the salt form is hydrochloride crystal form A, and its X-ray powder diffraction pattern has a diffraction angle 2θ are 6.9°±0.2°, 10.4°±0.2°, 13.9°±0.2°, 14.2°±0.2°, 15.5°±0.2°, 16.2°±0.2°, 16.9°±0.2°, 17.5°±0.2°, 18.1 The characteristic peaks are °±0.2°, 18.6°±0.2°, and 19.5°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 31 to 32, wherein the salt form is hydrochloride crystal form A, and its X-ray powder diffraction pattern is basically as follows As shown in Figure 27. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is hydrochloride crystal form I, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 7.5 The characteristic peaks are °±0.2°, 11.8°±0.2°, 16.7°±0.2°, 18.6°±0.2°, 18.9°±0.2°, and 21.8°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 34, wherein the salt form is hydrochloride crystal form I, and its X-ray powder diffraction pattern has a diffraction angle 2θ They are the characteristic peaks of 7.5°±0.2°, 10.7°±0.2°, 11.8°±0.2°, 16.7°±0.2°, 18.6°±0.2°, 18.9°±0.2°, and 21.8°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 34 to 35, wherein the salt form is hydrochloride crystal form I, and its X-ray powder diffraction pattern is basically as follows As shown in Figure 40. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is hydrochloride crystal form J, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 7.0 The characteristic peaks are °±0.2°, 10.5°±0.2°, 14.1°±0.2°, 17.6°±0.2°, 19.0°±0.2°, 24.9°±0.2°, and 26.1°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 37, wherein the salt form is hydrochloride crystal form J, and its X-ray powder diffraction pattern has a diffraction angle 2θ are 7.0°±0.2°, 10.5°±0.2°, 14.1°±0.2°, 17.6°±0.2°, 19.0°±0.2°, 21.4°±0.2°, 24.9°±0.2°, 26.1°±0.2°, 28.2 °±0.2° characteristic peak. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 37 to 38, wherein the salt form is hydrochloride crystal form J, and its X-ray powder diffraction pattern is basically as follows As shown in Figure 42. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is hydrochloride crystal form L, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 6.1 The characteristic peaks are °±0.2°, 11.4°±0.2°, 15.0°±0.2°, 15.4°±0.2°, and 18.9°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 40, wherein the salt form is hydrochloride crystal form L, and its X-ray powder diffraction pattern has a diffraction angle 2θ are 6.1°±0.2°, 11.4°±0.2°, 15.0°±0.2°, 15.4°±0.2°, 18.9°±0.2°, 21.4°±0.2°, 22.3°±0.2°, 23.0°±0.2°, 23.8 °±0.2° characteristic peak. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 40 to 41, wherein the salt form is hydrochloride crystal form L, and its X-ray powder diffraction pattern is basically as follows As shown in Figure 46. The salt form of the compound represented by formula I according to any one of claims 1 to 4, wherein the salt form is hydrochloride crystal form N, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 6.4 °±0.2°, 11.1°±0.2°, 12.8°±0.2°, 15.7°±0.2°, 16.2°±0.2°, 16.6°±0.2°, 18.7°±0.2°, 20.7°±0.2°, 22.1°± The characteristic peaks are 0.2°, 23.1°±0.2°, and 23.9°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 43, wherein the salt form is hydrochloride crystal form N, and its X-ray powder diffraction pattern has a diffraction angle 2θ are 6.4°±0.2°, 11.1°±0.2°, 11.4°±0.2°, 11.9°±0.2°, 12.8°±0.2°, 15.7°±0.2°, 16.2°±0.2°, 16.6°±0.2°, 18.7 The characteristic peaks are °±0.2°, 19.2°±0.2°, 20.7°±0.2°, 22.1°±0.2°, 23.1°±0.2°, and 23.9°±0.2°. The salt form of the compound represented by formula I as described in any one of claims 1 to 4 or 43 to 44, wherein the salt form is hydrochloride crystal form N, and its X-ray powder diffraction pattern is basically as follows As shown in Figure 50. A crystal form of a compound represented by formula I: Formula I. The crystal form of the compound represented by formula I as described in claim 46, wherein the crystal form is crystal form D, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 5.5°±0.2° and 14.7°±0.2 °, 15.4°±0.2°, 16.5°±0.2°, 17.5°±0.2°, and 21.7°±0.2°. The crystal form of the compound represented by formula I as described in claim 46 or 47, wherein the crystal form is crystal form D, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 5.5°±0.2° and 8.0° ±0.2°, 14.2°±0.2°, 14.7°±0.2°, 15.4°±0.2°, 16.5°±0.2°, 17.5°±0.2°, 18.0°±0.2°, 19.7°±0.2°, 21.7°±0.2 ° characteristic peaks. The crystal form of the compound represented by formula I according to any one of claims 46 to 48, wherein the crystal form is crystal form D, and its X-ray powder diffraction pattern is basically as shown in Figure 65. The crystal form of the compound represented by formula I as described in claim 46, wherein the crystal form is crystal form H, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 5.8°±0.2° and 13.0°±0.2 °, 14.3°±0.2°, 14.8°±0.2°, 16.3°±0.2°, 17.3°±0.2°, 17.9°±0.2°, and 21.9°±0.2°. The crystal form of the compound represented by formula I as described in claim 46 or 50, wherein the crystal form is crystal form H, and its X-ray powder diffraction pattern has a diffraction angle 2θ of 5.8°±0.2° and 7.4° ±0.2°, 13.0°±0.2°, 14.3°±0.2°, 14.8°±0.2°, 16.3°±0.2°, 17.3°±0.2°, 17.9°±0.2°, 21.9°±0.2°, 22.5°±0.2 ° characteristic peaks. The crystal form of the compound represented by formula I as described in any one of claims 46, 50 or 51, wherein the crystal form is crystal form H, and its X-ray powder diffraction pattern is basically as shown in Figure 73. An amorphous compound of formula I, wherein its X-ray powder diffraction pattern has no obvious sharp diffraction peaks; preferably, its X-ray powder diffraction pattern is basically as shown in Figure 79. A preparation method of a compound of formula I, which includes the following steps: Step 1: Compound S1 is prepared under the action of potassium acetate, water and acetic acid to obtain compound S2; Step 2: Compound S2 is under the action of oxidant potassium peroxyhydrosulfate complex salt to prepare compound S3; Step 3: Compound S3 is used and DHP as raw materials, in the presence of tetrahydrofuran and p-toluenesulfonic acid, prepare compound S21; Step 4: use compound S21 and compound S04 as raw materials, in the presence of acetonitrile and DIPEA, react to prepare compound S22; and step 5 : Compound S22 is first prepared under the action of hydrogen chloride-1,4-dioxane solution, and then under the action of sodium bicarbonate aqueous solution to prepare the compound of formula I. A method for preparing mesylate crystal form A as described in any one of claims 5 to 9, wherein it is prepared by the following steps: Using tetrahydrofuran/water as the solvent, the compound of formula I and methanesulfonic acid are stirred at a temperature of 40 to 80°C for 8 to 24 hours, filtered and dried to prepare the mesylate crystal form A of the compound of formula I. A method for preparing mesylate crystal form D as described in any one of claims 10 to 12, wherein it is prepared by the following steps: Using ethanol/water as a solvent, the compound of formula I and methanesulfonic acid are reacted at a temperature of 20 to 60°C for 0.5 to 3 hours. After filtration and drying, the mesylate crystal form D of the compound of formula I is prepared. A method for preparing mesylate crystal form A as described in any one of claims 5 to 9, wherein it is prepared by the following steps: j Use tetrahydrofuran/water as the solvent, stir and dissolve the compound of formula I at a temperature of 40 to 80°C, and add methanesulfonic acid with an equivalent of 0.1 to 0.3 relative to the compound of formula I; k While maintaining the reaction temperature, add the claim item 10 to 12 To the seed crystal of the mesylate crystal form D described in any one of the above, continue to add methanesulfonic acid with an equivalent of 0.7 to 0.9 relative to the compound of formula I, cool down, react for 1 to 12 hours, filter, and obtain crystal form D; l Mix the obtained mesylate crystal form D and the solvent at 40 to 80°C, then add the seed crystals of the methanesulfonate crystal form A described in any one of claims 5 to 9, and keep the reaction until the solid is completely Convert to mesylate crystal form A, cool down, and filter to prepare mesylate crystal form A. A pharmaceutical composition comprising the salt form of the compound represented by formula I as described in any one of claims 1 to 45 or the crystal form of the compound represented by formula I as described in any one of claims 46 to 52 Or the amorphous form of the compound represented by formula I as described in claim 53; and at least one pharmaceutically acceptable carrier or excipient. A salt form of the compound represented by formula I as described in any one of claims 1 to 45 or a crystal form of the compound represented by formula I as described in any one of claims 46 to 52 or as claimed in claim 53 The use of the amorphous form of the compound represented by formula I or the pharmaceutical composition as described in claim 58 in the preparation of medicines. The use as claimed in claim 59, wherein the medicine is used to treat, prevent, delay or prevent cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease. The use as claimed in claim 59, wherein the drug is used to treat diseases mediated by SHP2. The use as claimed in claim 61, wherein the disease is cancer. The use as claimed in claim 62, wherein the cancer is Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia , breast cancer, esophageal cancer, lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma and/or pancreatic cancer. A salt form of the compound represented by formula I as described in any one of claims 1 to 45 or a crystal form of the compound represented by formula I as described in any one of claims 46 to 52 or as claimed in claim 53 The use of the amorphous form of the compound represented by formula I or the pharmaceutical composition as described in claim 58 in the preparation of SHP2 inhibitors.