SHP2 inhibitors, crystalline forms thereof, and methods of making and using the same

Abstract

The application discloses SHP2 inhibitors, crystal forms thereof, and preparation methods and purposes thereof. The application discloses a compound B crystal form shown in formula (I), a compound shown in formula (II) and an A crystal form thereof, a compound shown in formula (III) and an A crystal form thereof, a compound shown in formula (IV) and an A crystal form thereof, a compound shown in formula (V) and an A crystal form thereof, and a compound shown in formula (VI) and an A crystal form thereof, and preparation methods and purposes thereof. The compound and the crystal form thereof have one or more advantages of stable property, good hygroscopicity, good bioavailability, and good drug-making prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to SHP2 inhibitors, crystal forms thereof, and preparation methods and uses thereof, and belongs to the field of chemical drugs. BACKGROUND

[0002] Protein tyrosine phosphatases (PTPs) play an important role in the regulation of a variety of cellular processes, such as cell growth, proliferation, cell differentiation, and oncogenic transformation. The balance between dephosphorylation by protein tyrosine phosphatases (PTPs) and phosphorylation by their counterpart tyrosine kinases is critical for normal physiological functions. PTPs are increasingly being recognized as valuable drug targets. For example, protein tyrosine phosphatase-2 (SHP2) encoded by tyrosine-protein phosphatase non-receptor type 11 (PTPN11) is a non-receptor protein tyrosine phosphatase containing two tandem Src homology-2 (SH2) domains. SHP2 is widely expressed in most tissues and plays a positive role in a variety of signal transduction pathways downstream of growth factor and cytokine receptors to regulate a variety of cellular functions. The catalytic activity of SHP2 is required for full activation of the Ras-ERK1 / 2 cascade, which is mediated by SHP2-catalyzed dephosphorylation of substrates that are negatively regulated by tyrosine phosphorylation. SHP2 is recognized as a true oncogene; gain-of-function SHP2 mutations lead to Noonan syndrome and various forms of leukemia (e.g., juvenile myelomonocytic leukemia, acute myeloid leukemia, myelodysplastic syndrome, acute lymphoblastic leukemia) and various solid tumors (e.g., lung adenocarcinoma, colon cancer, neuroblastoma, glioblastoma, melanoma, hepatocellular carcinoma, and prostate cancer). Therefore, SHP2 represents a promising target for a variety of cancers (e.g., triple-negative and HER2+ breast cancer, cancers resulting from abnormal activation of receptor protein tyrosine kinases (PTKs), some of which respond poorly to single therapy with kinase inhibitors), and is attracting increasing attention in the development of SHP2 inhibitors.

[0003] Therefore, it has gradually become a hot research field in industry and academia to discover and find SHP2 inhibitors with better drug properties.

[0004] The compound represented by formula (I) was first disclosed in PCT / CN2019 / 116386. The compound has a strong inhibitory effect on SHP2 and is highly selective, and is a new generation of SHP2 inhibitor.

[0005] SUMMARY

[0006] The present application aims to improve the physical and chemical properties of (S)-1'-(8-((((2-amino-3-chloropyridyl-4-yl)thio)-7-methylimidazo[1,2-c]pyrimidin-5-yl)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-amine (i.e., the compound shown in formula (I)) in the prior art, thereby providing a crystal form of (S)-1'-(8-((((2-amino-3-chloropyridyl-4-yl)thio)-7-methylimidazo[1,2-c]pyrimidin-5-yl)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-amine, a salt thereof, and a crystal form of the salt, as well as a preparation method and application thereof. The crystal form of (S)-1'-(8-((((2-amino-3-chloropyridyl-4-yl)thio)-7-methylimidazo[1,2-c]pyrimidin-5-yl)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-amine, the salt thereof, and each crystal form of the present application has one or more of the following advantages: better solubility, stable properties, good hygroscopicity, and good drug prospects.

[0007] The present application provides a compound shown in formula (II) or a crystal form thereof:

[0008]

[0009] In formula (II), M is citric acid, methanesulfonic acid, H2SO4, succinic acid, HCl, HNO3, HBr, HF, HI, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyhydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, octanoic acid, cinnamic acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyltartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galacturonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, 1,5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzene sulfonic acid, p-toluenesulfonic acid, and L-malic acid; preferably citric acid, methanesulfonic acid, H2SO4, succinic acid, HCl, HNO3, benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, malonic acid, and L-malic acid, ascorbic acid, salicylic acid, 2-acetoxybenzoic acid, nicotinic acid, isonicotinic acid, cholic acid, aspartic acid, or glutamic acid;

[0010] x is 0, 0.5, 1, 1.5, 2, 2.5, or 3;

[0011] y is 0, 1, 2, or 3;

[0012] x and y are not both 0.

[0013] In some embodiments, x is 0.5.

[0014] In some embodiments, x is 1.

[0015] In some embodiments, y is 0.

[0016] The present application provides a crystal form A of the compound of formula (I), which has an X-ray powder diffraction pattern substantially as shown in Figure 1

[0017]

[0018] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystal form A of the compound of formula (I) has endothermic peaks at 188.96 °C and 215.23 °C.

[0019] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystal form A of the compound of formula (I) has endothermic peaks at 188.96 °C and 215.23 °C. Figure 2

[0020] In some embodiments, the thermogravimetric analysis (TGA) curve of the crystal form A of the compound of formula (I) shows a weight loss of 0.21% from room temperature to 125 °C.

[0021] In some embodiments, the thermogravimetric analysis (TGA) curve of the crystal form A of the compound of formula (I) shows a weight loss of 0.21% from room temperature to 125 °C. Figure 3

[0022] The present application provides a crystal form B of the compound of formula (I), which has an X-ray powder diffraction pattern with characteristic diffraction peaks at the following 2Θ angles: 14.2 ± 0.2°, 12.53 ± 0.2°, 17.44 ± 0.2°, 17.76 ± 0.2°, 19.88 ± 0.2°, and 22.54 ± 0.2°.

[0023] ​​​In some embodiments, the X-ray powder diffraction pattern of the compound B Form of Formula (I) has characteristic diffraction peaks at the following 2Θ angles: 14.2 ± 0.2°, 12.53 ± 0.2°, 17.44 ± 0.2°, 17.76 ± 0.2°, 19.88 ± 0.2°, 22.54 ± 0.2°, 11.50 ± 0.2°, 16.52 ± 0.2°, 19.52 ± 0.2°, 20.17 ± 0.2°, 21.27 ± 0.2°, 23.44 ± 0.2°, 24.24 ± 0.2°, and 24.96 ± 0.2°.

[0024] In some embodiments, the X-ray powder diffraction pattern of the compound B Form of Formula (I) is substantially as shown in FIG. 1. Figure 4

[0025] In some embodiments, the X-ray powder diffraction pattern of the compound B Form of Formula (I) has characteristic diffraction peaks at the following 2Θ angles: 14.2 ± 0.2°, 12.53 ± 0.2°, 17.44 ± 0.2°, 17.76 ± 0.2°, 19.88 ± 0.2°, 22.54 ± 0.2°, 11.50 ± 0.2°, 16.52 ± 0.2°, 19.52 ± 0.2°, 20.17 ± 0.2°, 21.27 ± 0.2°, 23.44 ± 0.2°, 24.24 ± 0.2°, and 24.96 ± 0.2°.

[0026] Table 1: X-ray powder diffraction pattern of the compound B Form of Formula (I)

[0027]

[0028]

[0029] In some embodiments, the differential scanning calorimetry (DSC) curve of the compound B Form of Formula (I) has an endothermic peak at 207.09 °C.

[0030] In some embodiments, the differential scanning calorimetry (DSC) pattern of the compound B Form of Formula (I) is substantially as shown in FIG. 2. Figure 5

[0031] In some embodiments, the thermogravimetric analysis (TGA) curve of the compound B Form of Formula (I) shows a weight loss of 0.41% at room temperature to 112.39 °C, and a weight loss of 0.66% at room temperature to 224.70 °C.

[0032] In some embodiments, the thermogravimetric analysis (TGA) pattern of the compound B Form of Formula (I) is substantially as shown in FIG. 3. Figure 6

[0033] The present application provides a compound of Formula (III) (i.e., a mesylate salt of a compound of Formula (I)), a crystalline form or a hydrate thereof;

[0034]

[0035] ​​​The present application provides a crystal form A of a compound of Formula (III), wherein the crystal form has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 10.2 ± 0.2°, 12.2 ± 0.2°, 15.9 ± 0.2°, 17.2 ± 0.2°, 18.6 ± 0.2°, and 19.5 ± 0.2°.

[0036] In some embodiments, the crystal form A of the compound of Formula (III) has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 10.2 ± 0.2°, 11.0 ± 0.2°, 12.2 ± 0.2°, 13.4 ± 0.2°, 13.9 ± 0.2°, 14.4 ± 0.2°, 15.9 ± 0.2°, 16.9 ± 0.2°, 17.2 ± 0.2°, 18.6 ± 0.2°, 19.5 ± 0.2°, 20.2 ± 0.2°, 20.7 ± 0.2°, 21.5 ± 0.2°, 22.5 ± 0.2°, 22.9 ± 0.2°, 24.5 ± 0.2°, 25.0 ± 0.2°, 25.5 ± 0.2°, 27.2 ± 0.2°, 28.6 ± 0.2°, 28.7 ± 0.2°, 29.6 ± 0.2°, 30.1 ± 0.2°, and 30.5 ± 0.2°.

[0037] In some embodiments, the crystal form A of the compound of Formula (III) has an X-ray powder diffraction pattern substantially as depicted in Figure 7

[0038] In some embodiments, the crystal form A of the compound of Formula (III) has an X-ray powder diffraction pattern with characteristic peaks at 2-theta values of 10.2 ± 0.2°, 12.2 ± 0.2°, 15.9 ± 0.2°, 17.2 ± 0.2°, 18.6 ± 0.2°, and 19.5 ± 0.2°.

[0039] Table 2: X-ray powder diffraction pattern of the crystal form A of the compound of Formula (III)

[0040]

[0041]

[0042] In some embodiments, the crystal form A of the compound of Formula (III) has a differential scanning calorimetry (DSC) profile with endothermic peaks at 50.39 °C and 204.24 °C.

[0043] In some embodiments, the crystal form A of the compound of Formula (III) has a differential scanning calorimetry (DSC) profile substantially as depicted in Figure 8

[0044] In some embodiments, the crystal form A of the compound of Formula (III) has a thermogravimetric analysis (TGA) profile showing a weight loss of 3.5% from room temperature to 87 °C. In some embodiments, the crystal form A of the compound of Formula (III) has a differential scanning calorimetry (DSC) profile with endothermic peaks at 50.39 °C and 204.24 °C.​

[0045] In some embodiments, the thermal gravimetric analysis (TGA) pattern of the crystalline form of the compound of Formula (III) A is as shown in Figure 9 .

[0046] The present application provides a compound of Formula (IV) (i.e., a citrate salt of the compound of Formula (I)), a crystalline form or a hydrate thereof;

[0047]

[0048] The present application provides a crystalline form of the compound of Formula (IV) A, wherein the crystalline form has an X-ray powder diffraction pattern with characteristic peaks at the following 2Θ angles: 9.5±0.2°, 11.5±0.2°, 11.8±0.2°, 3.5±0.2°, 14.1±0.2°, 16.3±0.2°, 18.0±0.2° 20.7±0.2° and 25.1±0.2°.

[0049] In some embodiments, the X-ray powder diffraction pattern of the crystalline form of the compound of Formula (IV) A has characteristic peaks at the following 2Θ angles: 9.5±0.2°, 10.3±0.2°, 11.5±0.2°, 11.8±0.2°, 12.2±0.2°, 12.9±0.2°, 13.5±0.2°, 14.1±0.2°, 15.7±0.2°, 16.3±0.2°, 17.0±0.2°, 18.0±0.2°, 18.6±0.2°, 20.7±0.2°, 21.1±0.2°, 22.2±0.2°, 23.2±0.2°, 23.8±0.2°, 24.5±0.2°, 24.8±0.2°, 25.1±0.2°, 26.2±0.2°, 28.7±0.2°, 29.4±0.2°, 29.7±0.2°, 30.5±0.2°, 31.8±0.2°, 32.3±0.2°, 33.9±0.2°, 34.6±0.2°, 35.4±0.2°, 36.5±0.2° and 40.4±0.2°.

[0050] In some embodiments, the X-ray powder diffraction pattern of the crystalline form of the compound of Formula (IV) A is substantially as shown in Figure 10 .

[0051] In some embodiments, the X-ray powder diffraction pattern of the crystalline form of the compound of Formula (IV) A has characteristic peaks at the following 2Θ angles: 9.5±0.2°, 10.3±0.2°, 11.5±0.2°, 11.8±0.2°, 12.2±0.2°, 12.9±0.2°, 13.5±0.2°, 14.1±0.2°, 15.7±0.2°, 16.3±0.2°, 17.0±0.2°, 18.0±0.2°, 18.6±0.2°, 20.7±0.2°, 21.1±0.2°, 22.2±0.2°, 23.2±0.2°, 23.8±0.2°, 24.5±0.2°, 24.8±0.2°, 25.1±0.2°, 26.2±0.2°, 28.7±0.2°, 29.4±0.2°, 29.7±0.2°, 30.5±0.2°, 31.8±0.2°, 32.3±0.2°, 33.9±0.2°, 34.6±0.2°, 35.4±0.2°, 36.5±0.2° and 40.4±0.2°.

[0052] Table 3: X-ray powder diffraction pattern resolution data of the crystalline form of the compound of Formula (IV) A

[0053]

[0054]

[0055] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystalline form A of the compound of Formula (IV) has endothermic peaks at 90.14 °C, 162.81 °C and 189.67 °C.

[0056] In some embodiments, the differential scanning calorimetry (DSC) profile of the crystalline form A of the compound of Formula (IV) is as shown in Figure 11

[0057] In some embodiments, the thermogravimetric analysis (TGA) curve of the crystalline form A of the compound of Formula (IV) shows a weight loss of 3.64% from room temperature to 115 °C.

[0058] In some embodiments, the thermogravimetric analysis (TGA) profile of the crystalline form A of the compound of Formula (IV) is as shown in Figure 12

[0059] The present application provides a compound of Formula (V) (i.e. a sulfate salt of the compound of Formula (I)), a crystalline form or a hydrate thereof;

[0060]

[0061] The present application provides a crystalline form A of the compound of Formula (V), wherein the X-ray powder diffraction pattern of the crystalline form has characteristic diffraction peaks at the following 2Θ angles: 10.02±0.2°, 16.06±0.2°, 16.58±0.2°, 21.96±0.2°, 24.38±0.2° and 24.96±0.2°.

[0062] In some embodiments, the X-ray powder diffraction pattern of the crystalline form A of the compound of Formula (V) has characteristic diffraction peaks at the following 2Θ angles: 9.70±0.2°, 10.02±0.2°, 10.80±0.2°, 11.84±0.2°, 13.38±0.2°, 14.14±0.2°, 15.18±0.2°, 14.1±0.2°, 16.06±0.2°, 16.58±0.2°, 17.16±0.2°, 18.36±0.2°, 19.54±0.2°, 21.96±0.2°, 22.40±0.2°, 23.80±0.2°, 24.38±0.2°, 24.96±0.2°, 27.02±0.2°, 27.63±0.2°, 28.74±0.2°, 30.30±0.2°, 32.08±0.2°, 33.67±0.2° and 34.47±0.2°.

[0063] ​​In some embodiments, the X-ray powder diffraction pattern of the crystalline form of the compound of Formula (V) is substantially as shown in FIG. 2. Figure 13

[0064] In some embodiments, the X-ray powder diffraction pattern of the crystalline form of the compound of Formula (V) is substantially as shown in FIG. 2.

[0065] Table 4: X-ray powder diffraction pattern data of the crystalline form of the compound of Formula (V)

[0066]

[0067] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystalline form of the compound of Formula (V) has endothermic peaks at 51.72 °C and 223 °C.

[0068] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystalline form of the compound of Formula (V) has endothermic peaks at 51.72 °C and 223 °C. Figure 14

[0069] In some embodiments, the thermogravimetric analysis (TGA) curve of the crystalline form of the compound of Formula (V) shows a weight loss of 3.37% from room temperature to 85 °C.

[0070] In some embodiments, the thermogravimetric analysis (TGA) curve of the crystalline form of the compound of Formula (V) shows a weight loss of 3.37% from room temperature to 85 °C. Figure 15

[0071] The present application provides a compound of Formula (VI) (i.e., a succinate salt of the compound of Formula (I)), a crystalline form or a hydrate thereof;

[0072]

[0073] The present application provides a crystalline form of the compound of Formula (VI), wherein the X-ray powder diffraction pattern of the crystalline form has characteristic diffraction peaks at the following 2Θ angles: 317.32 ± 0.2°, 18.16 ± 0.2°, 20.62 ± 0.2°, 20.86 ± 0.2°, 22.46 ± 0.2°, 24.00 ± 0.2°, 24.34 ± 0.2°, and 25.02 ± 0.2°.

[0074] ​​​In some embodiments, the X-ray powder diffraction pattern of the A-type of the compound shown in formula (VI) has characteristic diffraction peaks at the following 2θ angles: 9.10±0.2°, 11.06±0.2°, 11.46±0.2°, 13.46±0.2°, 14.34±0.2°, 15.50±0.2°, 16.63±0.2°, 16.96±0.2°, 17.32±0.2°, and 18.16±0.2°. 2°, 19.08±0.2°, 20.62±0.2°, 20.86±0.2°, 22.46±0.2°, 23.36±0.2°, 24.00±0.2°, 24.34±0.2°, 25.02±0.2°, 25.92±0.2°, 26.28±0.2°, 27.84±0.2°, 28.10±0.2°, 28.88±0.2°, 30.45±0.2°.

[0075] In some embodiments, the X-ray powder diffraction pattern of the A-crystal form of the compound shown in formula (VI) is substantially as follows: Figure 16 As shown.

[0076] In some embodiments, the X-ray powder diffraction pattern analysis data of the A-phase of the compound represented by formula (VI) are shown in Table 5:

[0077] Table 5: X-ray powder diffraction pattern analysis data of the A-crystal form of the compound shown in formula (VI)

[0078]

[0079]

[0080] In some embodiments, the differential scanning calorimetry (DSC) curve of the A crystal form of the compound shown in formula (VI) has an endothermic peak at 164.53 °C.

[0081] In some embodiments, the differential scanning calorimetry (DSC) spectrum of the A-phase of the compound represented by formula (VI) is as follows: Figure 17 As shown.

[0082] In some embodiments, the thermogravimetric analysis (TGA) curves of the A-form of the compound shown in formula (VI) show a weight loss of 1.42% from room temperature to 100°C.

[0083] In some embodiments, the thermogravimetric analysis (TGA) spectrum of the A-phase of the compound represented by formula (VI) is as follows: Figure 18 As shown.

[0084] In some implementations, the purity of the crystal form is above 95%.

[0085] The application provides a preparation method of a compound shown in formula (II), which comprises the following steps: performing salt reaction on a compound shown in formula (I) and an acid in a solvent to obtain a compound shown in formula (II). In formula (II), the M is citric acid, methanesulfonic acid, H2SO4, succinic acid, HCl, HNO3, HBr, HF, HI, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphthoic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyhydroxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetylamino benzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, octanoic acid, cinnamic acid, cyclohexanesulfamic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, dodecylsulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galacturonic acid, gentisic acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, isethionic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, 1,5-naphthalene disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, thiocyanic acid, undecylenic acid, trifluoroacetic acid, benzene sulfonic acid, p-toluenesulfonic acid and L-malic acid; preferably citric acid, methanesulfonic acid, H2SO4, succinic acid, HCl, HNO3, benzenesulfonic acid, maleic acid, adipic acid, p-toluenesulfonic acid, malonic acid and L-malic acid, ascorbic acid, salicylic acid, 2-acetoxybenzoic acid, nicotinic acid, isonicotinic acid, cholic acid, aspartic acid or glutamic acid.

[0086] In some embodiments, the solvent is one or more of halogenated hydrocarbons, dioxane, nitriles, alcohols and water. Preferably, it is one or more of DCM, acetonitrile, dioxane, water and ethanol.

[0087] The application provides a preparation method of a compound shown in formula (III), which comprises the following steps: performing salt reaction on a compound shown in formula (I) and an acid in a solvent to obtain a compound shown in formula (III). In the formula, the acid is methanesulfonic acid. The solvent is dioxane.

[0088] The application provides a preparation method of a compound shown in formula (IV), which comprises the following steps: performing salt reaction on a compound shown in formula (I) and an acid in a solvent to obtain a compound shown in formula (IV). In the formula, the acid is citric acid. The solvent is a mixture solution of acetonitrile and water.

[0089] The application provides a preparation method of a compound shown as formula (V), which comprises the following steps: carrying out salt formation reaction of a compound shown as formula (I) and an acid in a solvent to obtain the compound shown as formula (V). Wherein the acid is sulfuric acid, and the solvent is ethanol.

[0090] The application provides a preparation method of a compound shown as formula (VI), which comprises the following steps: carrying out salt formation reaction of a compound shown as formula (I) and an acid in a solvent to obtain the compound shown as formula (VI). Wherein the acid is succinic acid, and the solvent is dioxane.

[0091] The application provides a preparation method of a compound A shown as formula (I), which comprises the following steps: carrying out the following reaction of a compound shown as formula H in DCM and TFA, then extracting with DCM, drying the DCM phase to obtain the compound A shown as formula (I).

[0092]

[0093] In some embodiments, in the preparation method of the compound A shown as formula (I), the mass / volume ratio of the compound shown as formula H and DCM can be 15-45 mg / mL, preferably 30 mg / mL.

[0094] In some embodiments, in the preparation method of the compound A shown as formula (I), the mass / volume ratio of the compound shown as formula H and TFA can be 100-200 mg / mL, preferably 150 mg / mL.

[0095] In some embodiments, in the preparation method of the compound A shown as formula (I), the extraction operation is adding DCM and saturated Na2CO3 aqueous solution to the reaction solution, and separating the DCM phase.

[0096] In some embodiments, in the preparation method of the compound A shown as formula (I), the drying condition is 30-40 DEG C. Preferably, 30-40 DEG C. rotary drying.

[0097] In some embodiments, in the preparation method of the compound A shown as formula (I), the reaction is carried out at room temperature.

[0098] In some embodiments, in the preparation method of the compound A shown as formula (I), the specific operation of the preparation method of the compound A shown as formula (I) is: dissolving the compound shown as formula H in DCM, adding TFA at room temperature to carry out the reaction, concentrating the reaction solution, adding DCM for dilution, adding saturated Na2CO3 aqueous solution, extracting, and taking the DCM phase to rotary dry at 30-40 DEG C.

[0099] The application provides a preparation method of a compound B crystal form shown in formula (I), which comprises the following steps: crystallizing compound A crystal form shown in formula (I) after mixing with acetonitrile to obtain the compound B crystal form shown in formula (I).

[0100] In some embodiments, in the preparation method of the compound B crystal form shown in formula (I), the mass / volume ratio of the compound A crystal form shown in formula (I) and acetonitrile can be 10-60 mg / mL. Preferably, the mass / volume ratio is 30 mg / mL.

[0101] In some embodiments, in the preparation method of the compound B crystal form shown in formula (I), the mixing operation can be shaking table shaking. Preferably, the shaking table shaking is carried out at 25 DEG C. More preferably, the shaking table shaking is carried out at 25 DEG C and 250 rpm. Further preferably, the shaking table shaking is carried out at 25 DEG C and 250 rpm for 24 h.

[0102] In some embodiments, in the preparation method of the compound B crystal form shown in formula (I), the crystallization operation can comprise the following steps: separating the solid in the system after mixing the compound A crystal form shown in formula (I) with acetonitrile to obtain the compound B crystal form shown in formula (I). Preferably, the separated solid is further dried. More preferably, the drying is carried out at 50 DEG C. Further preferably, the drying is carried out at 50 DEG C and a vacuum degree of -0.1 M. Further preferably, the drying is carried out at 50 DEG C and a vacuum degree of -0.1 M for 6 h. Further preferably, the drying is carried out in a vacuum drying box at 50 DEG C and a vacuum degree of -0.1 M for 6 h.

[0103] The application provides a preparation method of a compound A crystal form shown in formula (III), which comprises the following steps: mixing the compound A crystal form shown in formula (I) with dioxane, and then reacting with a methyl methanesulfonate methanol solution to crystallize to obtain the compound A crystal form shown in formula (III).

[0104] In some embodiments, in the preparation method of the compound A crystal form shown in formula (III), the mixing operation can be stirring. Preferably, the stirring is magnetic stirring while the temperature is increased to 50 DEG C.

[0105] In some embodiments, in the preparation method of the compound A crystal form shown in formula (III), the mass / volume ratio of the compound A crystal form shown in formula (I) and dioxane can be 20-90 mg / mL. Preferably, the mass / volume ratio is 50 mg / mL.

[0106] In some embodiments, in the preparation method of the compound A crystal form shown in formula (III), the mass / volume ratio of the compound A crystal form shown in formula (I) and the methyl methanesulfonate methanol solution can be 200-900 mg / mL. Preferably, the mass / volume ratio is 500 mg / mL.

[0107] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), the concentration of the methyl methanesulfonate methanol solution can be 0.5-2 mol / L. Preferably, the concentration of the methyl methanesulfonate methanol solution is 1 mol / L.

[0108] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), the operation of reaction and crystallization can comprise the following steps: after mixing the compound A crystalline form of formula (I) and the methyl methanesulfonate methanol solution, cooling, and separating the solid in the mixture after reaction.

[0109] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), the reaction can be carried out at a temperature of 50°C. Preferably, the reaction can be carried out at a temperature of 50°C for 3 h.

[0110] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), the operation of cooling can comprise cooling to room temperature at a cooling rate of 10°C / h, and then placing in a 4°C refrigerator for 24 h.

[0111] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), after the solid is separated out, the solid can be further washed. Preferably, the solid is washed with a dioxane solution. More preferably, the solid is washed with a 4°C dioxane solution. Further preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to the dioxane solution is 150 mg / mL.

[0112] In some embodiments, in the preparation method of the compound A crystalline form of formula (III), after the solid is separated out, the solid can be further dried. Preferably, the solid is dried at 50°C. Further preferably, the solid is dried at 50°C under a vacuum degree of -0.1 M. Further preferably, the solid is dried at 50°C under a vacuum degree of -0.1 M for 24 h. Further preferably, the solid is dried in a vacuum drying oven at 50°C under a vacuum degree of -0.1 M for 24 h.

[0113] The present application provides a preparation method of a compound A crystalline form of formula (IV), which comprises the following steps: mixing the compound A crystalline form of formula (I) with acetonitrile and purified water, and then reacting with a methyl citrate solution to crystallize, to obtain the compound A crystalline form of formula (IV).

[0114] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the operation of mixing can be stirring. Preferably, the operation of mixing is stirring while heating to 50°C. More preferably, the operation of mixing is stirring (200-300 rpm) while heating to 50°C.

[0115] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the mass / volume ratio of the compound A crystalline form of formula (I) to acetonitrile can be 5-25 mg / mL. Preferably, 11.1 mg / mL.

[0116] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the mass / volume ratio of the compound A crystalline form of formula (I) to purified water can be 30-170 mg / mL. Preferably, 100 mg / mL.

[0117] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the mass / volume ratio of the compound A crystalline form of formula (I) to the methanol solution of citric acid can be 200-900 mg / mL. Preferably, 500 mg / mL.

[0118] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the concentration of the methanol solution of citric acid can be 0.5-2 mol / L. Preferably, 1 mol / L.

[0119] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the operation of reaction crystallization can include the following steps: after mixing the compound A crystalline form of formula (I) with acetonitrile, purified water and the methanol solution of citric acid, cooling, and separating the solid from the mixture after reaction.

[0120] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the reaction can be carried out at a temperature of 50°C. Preferably, the reaction can be carried out by stirring at a temperature of 50°C. More preferably, the reaction can be carried out by stirring (rotation speed 200-300 rpm) at a temperature of 50°C for 2 h.

[0121] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), the cooling operation can include cooling to 5°C at a cooling rate of 15°C / h. Preferably, cooling to 5°C at a cooling rate of 15°C / h, and then crystallization for 0.5 h.

[0122] In some embodiments, in the preparation method of the compound A crystalline form of formula (IV), after the solid is separated, it can be further washed. Preferably, acetonitrile solution is used for washing. More preferably, 4°C acetonitrile solution is used for washing. Further preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to acetonitrile solution is 150 mg / mL.

[0123] In some embodiments, the method for preparing the compound A crystalline form of formula (IV) can further comprise a drying step after the solid is separated out. Preferably, the drying is performed at 50°C. Further preferably, the drying is performed at 50°C under a vacuum of -0.1 M. Further preferably, the drying is performed at 50°C under a vacuum of -0.1 M for 24 h. Further preferably, the drying is performed at 50°C under a vacuum of -0.1 M in a vacuum drying oven for 24 h.

[0124] The present application provides a method for preparing a compound A crystalline form of formula (V), which comprises the following steps: mixing a compound A crystalline form of formula (I) with ethanol, and then reacting with a methanol solution of sulfuric acid to crystallize, thereby obtaining a compound A crystalline form of formula (V).

[0125] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0126] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0127] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0128] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0129] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0130] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise a heating step after the mixing, in which the temperature is raised to 50°C.

[0131] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise the following steps: cooling the reaction solution to 25°C at a cooling rate of 12.5°C / h, adding acetonitrile, and cooling the reaction solution to 5°C at a cooling rate of 10°C / h. Preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to acetonitrile can be 10 mg / mL.

[0132] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise the following steps: cooling the reaction solution to 25°C at a cooling rate of 12.5°C / h, adding acetonitrile, and cooling the reaction solution to 5°C at a cooling rate of 10°C / h. Preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to acetonitrile can be 10 mg / mL.

[0133] In some embodiments, the method for preparing the compound A crystalline form of formula (V) can further comprise the following steps: cooling the reaction solution to 25°C at a cooling rate of 12.5°C / h, adding acetonitrile, and cooling the reaction solution to 5°C at a cooling rate of 10°C / h. Preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to acetonitrile can be 10 mg / mL.

[0134] The present application provides a method for preparing a compound A crystalline form of formula (VI), which comprises the following steps: mixing the compound A crystalline form of formula (I) with dioxane, and then reacting with a methanol solution of succinic acid to crystallize, thereby obtaining the compound A crystalline form of formula (VI).

[0135] In some embodiments, the method for preparing the compound A crystalline form of formula (VI) can further comprise the following steps: mixing the compound A crystalline form of formula (I) with dioxane, and then reacting with a methanol solution of succinic acid to crystallize, thereby obtaining the compound A crystalline form of formula (VI). Preferably, the mixing can be stirring. More preferably, the stirring can be magnetic stirring while heating to 50°C.

[0136] In some embodiments, the method for preparing the compound A crystalline form of formula (VI) can further comprise the following steps: mixing the compound A crystalline form of formula (I) with dioxane, and then reacting with a methanol solution of succinic acid to crystallize, thereby obtaining the compound A crystalline form of formula (VI). Preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to dioxane can be 15-70 mg / mL. More preferably, the mass / volume ratio can be 37.5 mg / mL.

[0137] In some embodiments, the method for preparing the compound A crystalline form of formula (VI) can further comprise the following steps: mixing the compound A crystalline form of formula (I) with dioxane, and then reacting with a methanol solution of succinic acid to crystallize, thereby obtaining the compound A crystalline form of formula (VI). Preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to the methanol solution of succinic acid can be 200-900 mg / mL. More preferably, the mass / volume ratio can be 500 mg / mL.

[0138] In some embodiments, the method for preparing the compound A crystalline form of formula (VI) can further comprise the following steps: mixing the compound A crystalline form of formula (I) with dioxane, and then reacting with a methanol solution of succinic acid to crystallize, thereby obtaining the compound A crystalline form of formula (VI). Preferably, the concentration of the methanol solution of succinic acid can be 0.5-2 mol / L. More preferably, the concentration can be 1 mol / L.

[0139] In some embodiments, in the method for preparing the compound A crystalline form of formula (VI), the operation of reaction crystallization can comprise the following steps: after mixing the compound A crystalline form of formula (I) with dioxane and reacting with the methanol solution of succinic acid, cooling, and separating the solid from the mixture after reaction.

[0140] In some embodiments, in the method for preparing the compound A crystalline form of formula (VI), the reaction can be carried out at a temperature of 50°C. Preferably, the reaction can be carried out at a temperature of 50°C for 3h.

[0141] In some embodiments, in the method for preparing the compound A crystalline form of formula (VI), the operation of cooling can comprise cooling to room temperature at a cooling rate of 10°C / h, and then placing in a 4°C refrigerator for 5h.

[0142] In some embodiments, in the method for preparing the compound A crystalline form of formula (VI), after the solid is separated, it can be further washed. Preferably, it is washed with a dioxane solution. More preferably, it is washed with a 4°C dioxane solution. Further preferably, the mass / volume ratio of the compound A crystalline form of formula (I) to the dioxane solution is 150mg / mL.

[0143] In some embodiments, in the method for preparing the compound A crystalline form of formula (VI), after the solid is separated, it can be further dried. Preferably, it is dried at 50°C. Further preferably, it is dried at 50°C under a vacuum degree of -0.1M. Further preferably, it is dried at 50°C under a vacuum degree of -0.1M for 24h. Further preferably, it is dried at 50°C under a vacuum degree of -0.1M in a vacuum drying oven for 24h.

[0144] The present application further provides a pharmaceutical composition containing a therapeutically effective amount of any one of the compounds or crystalline forms described in the present application, and a pharmaceutically acceptable excipient.

[0145] In some embodiments, the pharmaceutical composition is for oral administration.

[0146] In some embodiments, the pharmaceutical composition is for making tablets or capsules.

[0147] In some embodiments, the pharmaceutical composition contains 0.2-10% by weight of any one of the compound crystalline forms described in the present application.

[0148] The present application further provides the use of any one of the compounds or crystalline forms or the pharmaceutical composition in the preparation of a medicament.

[0149] In some embodiments, the medicament is a medicament for treating, preventing, delaying or hindering the occurrence or progression of a disease associated with the activity or expression of SHP2 protein.

[0150] In some embodiments, the medicament is a medicament for treating a disease associated with SHP2 protein activity or expression.

[0151] In some embodiments, the disease is a tumor.

[0152] In some embodiments, the tumor is a tumor caused by abnormality of Ras-Raf-ERK or PD1 / L1 signaling pathway.

[0153] In some embodiments, the tumor is esophageal cancer, lung cancer, colorectal cancer, pancreatic cancer, leukemia or gastric cancer.

[0154] In the present application, "about" and "substantially" used in "having an X-ray powder diffraction pattern about as shown in the figure" or "the powder diffraction pattern thereof is substantially as shown in the figure" means that the exact position of the peaks in the figure should not be interpreted as absolute value. Because it is known to those skilled in the art that the 2θ value of the X-ray powder diffraction pattern can be subject to error under different measurement conditions (such as the equipment and instruments used) and different samples, the measurement error of the diffraction angle of the X-ray powder diffraction pattern is 5% or less, and generally, the difference of ±0.2° of a given value is considered appropriate. It should also be understood that the relative intensity of the peak value can fluctuate with experimental conditions and sample preparation such as the preferred orientation of particles in the sample. The use of automatic or fixed divergence slits will also affect the calculation of relative intensity. The intensity shown in the PXRD curve included herein is only exemplary and cannot be used as an absolute comparison.

[0155] Those skilled in the art should understand that the data measured by DSC can have small changes due to changes in sample purity, sample preparation, and measurement conditions (such as heating rate). Alternative melting point readings can be given by other kinds of instruments or using conditions different from those described herein. Therefore, the endothermic pattern used in the present application cannot be taken as an absolute value, and such measurement errors should be considered when interpreting the DSC data.

[0156] The starting temperature of the thermogravimetric analysis (TGA) test is not specified in the present application, and the starting temperature thereof is room temperature, which is generally 20-35°C.

[0157] The term "therapeutically effective amount" as used herein refers to the amount of a compound that, when administered to a subject for treatment of a disease, or at least one of the clinical symptoms of a disease or condition, is sufficient to affect such treatment for the disease, condition, or symptom. The "therapeutically effective amount" will vary depending on the compound, the disease, condition, and / or symptoms of the disease or condition, the severity of the disease, condition, and / or symptoms of the disease or condition, the age of the patient, and / or the body weight of the patient, etc. An appropriate amount in any particular case will be readily ascertainable by those skilled in the art, or can be determined by routine experimentation. In the case of combination therapy, the "therapeutically effective amount" refers to the total amount of the combination administered to the subject.

[0158] The salt forms or crystal forms described herein can be combined with a pharmaceutical carrier selected on the basis of conventional pharmaceutical practice, to prepare pharmaceutical compositions. The pharmaceutical carrier can take a wide variety of forms depending on the form of administration (e.g., oral or injection (including intravenous injection)). Thus, the pharmaceutical compositions of the present application can take the form of tablets, capsules, pills, granules, powders, solutions, suspensions, or aerosols, as examples, for oral administration. Further, the pharmaceutical compositions of the present application can take the form of a liquid solution, a liquid emulsion, or a liquid suspension. In addition, the salt forms or crystal forms described herein can be administered by controlled release and / or delivery devices. The pharmaceutical compositions of the present application can be manufactured in any manner which is known to the art. Generally, such methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more necessary ingredients. In general, the pharmaceutical compositions are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. Additionally, the product can be conveniently formulated into a desired shape.

[0159] The term "pharmaceutically acceptable carrier" as used herein means a conventional pharmaceutical carrier suitable for the desired pharmaceutical preparation, for example: diluents such as water, various organic solvents, and the like; excipients; fillers such as starch, pregelatinized starch, sucrose, dextrin, mannitol, lactose, spray-dried lactose, microcrystalline cellulose, silicified microcrystalline cellulose, inorganic salts, and the like; binders such as starch paste, dextrin, powdered sugar, sugar syrup, gelatin, hydroxypropyl cellulose, copovidone, and polyvinylpyrrolidone (PVP); humectants such as distilled water, ethanol, and glycerol; disintegrants such as dry starch, low-substituted hydroxypropyl cellulose, hydroxypropyl starch, agar, calcium carbonate, sodium bicarbonate, cross-linked polyplasdone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, and the like; absorption enhancers such as quaternary ammonium compounds, amino acid ethylamine derivatives, acetoacetate esters, β-dicarboxylic acid esters, aromatic acidic compounds, aliphatic acidic compounds, and the like; surfactants such as sodium lauryl sulfate, sodium octadecyl sulfate, dioctyl sodium sulfosuccinate, sodium laurylsulfate, benzalkonium bromide, benzalkonium chloride, dodecylamine, lecithin, cetyl alcohol, sodium dodecyl sulfate, Tween, and Span, and the like; drug- carrying bases such as polyethylene glycol, carbomer, cellulose derivatives, glycerogelatin, polyvinyl alcohol, cocoa butter, synthetic or wholly synthetic fatty acid glycerides, polyvinyl alcohol 40 stearate, petrolatum, solid paraffin, liquid paraffin, dimethyl silicone oil, lanolin, beeswax, and lanolin ester, and the like; absorption carriers such as kaolin and bentonite, and the like; lubricants such as talc, micronized silica gel, silicon dioxide, hydrogenated vegetable oil, magnesium lauryl sulfate, sodium lauryl sulfate, stearic acid, calcium stearate, magnesium stearate, sodium stearyl fumarate, and polyethylene glycol, and the like. In addition, other pharmaceutically acceptable adjuvants such as antioxidants, colorants, preservatives, pH adjusters, hardening agents, emulsifiers, propellants, dispersants, stabilizers, thickening agents, complexing agents, buffers, penetration enhancers, polymers, fragrances, sweeteners, and dyes can be added to the pharmaceutical composition. Preferably, the adjuvants suitable for the desired dosage form and the desired administration method are used.

[0160] On the basis of common knowledge in the art, the above-mentioned preferred conditions can be combined arbitrarily, i.e. to obtain each preferred example of the present application.

[0161] The reagents and raw materials used in the present application are commercially available.

[0162] The positive progress effect of the present application is that the compound A, B crystal form represented by formula (I), the compound A crystal form represented by formula (III), the compound A crystal form represented by formula (IV), the compound A crystal form represented by formula (V), and the compound A crystal form represented by formula (VI) protected in the present application have one or more of the following advantages: (1) stable properties; (2) good hygroscopicity; (3) good bioavailability; (4) good drug development prospects. BRIEF DESCRIPTION OF DRAWINGS

[0163] Figure 1 : PXRD pattern of Compound A Form A of Formula (I) - X-ray powder diffraction pattern.

[0164] Figure 2 : DSC pattern of Compound A Form D of Formula (I) - Differential scanning calorimetry pattern.

[0165] Figure 3 : TGA pattern of Compound A Form D of Formula (I) - Thermogravimetric analysis pattern.

[0166] Figure 4 : PXRD pattern of Compound B Form of Formula (I).

[0167] Figure 5 : DSC pattern of Compound B Form of Formula (I).

[0168] Figure 6 : TGA pattern of Compound B Form of Formula (I).

[0169] Figure 7 : PXRD pattern of Compound A Form of Formula (III).

[0170] Figure 8 : DSC pattern of Compound A Form of Formula (III).

[0171] Figure 9 : TGA pattern of Compound A Form of Formula (III).

[0172] Figure 10 : PXRD pattern of Compound A Form of Formula (IV).

[0173] Figure 11 : DSC pattern of Compound A Form of Formula (IV).

[0174] Figure 12 : TGA pattern of Compound A Form of Formula (IV).

[0175] Figure 13 : PXRD pattern of Compound A Form of Formula (V).

[0176] Figure 14 : DSC pattern of Compound A Form of Formula (V).

[0177] Figure 15 : TGA pattern of Compound A Form of Formula (V).

[0178] Figure 16 : PXRD pattern of Compound A Form of Formula (VI).

[0179] Figure 17DSC pattern of compound A in form of formula (VI) crystal.

[0180] Figure 18 TGA pattern of compound A in form of formula (VI) crystal.

[0181] Figure 19 H NMR pattern of compound B in form of formula (I) crystal. 1 NMR pattern.

[0182] Figure 20 H NMR pattern of compound A in form of formula (III) crystal. 1 NMR pattern.

[0183] Figure 21 H NMR pattern of compound A in form of formula (IV) crystal. 1 NMR pattern.

[0184] Figure 22 H NMR pattern of compound A in form of formula (V) crystal. 1 NMR pattern.

[0185] Figure 23 H NMR pattern of compound A in form of formula (VI) crystal. 1 NMR pattern.

[0186] Figure 24 Effect of each test substance on tumor volume of human non-small cell lung cancer NCI-H358 nude mouse xenograft model animal.

[0187] Figure 25 Effect of each test substance on tumor weight of human non-small cell lung cancer NCI-H358 nude mouse xenograft model animal.

[0188] Figure 26 Effect of each test substance on tumor volume of human leukemia MV-4-11 mouse xenograft model animal.

[0189] Figure 27 Effect of each test substance on tumor weight of human leukemia MV-4-11 mouse xenograft model animal.

[0190] Figure 28 Effect of each test substance on tumor volume of human pancreatic cancer Mia PaCa-2 nude mouse xenograft model animal.

[0191] Figure 29 Effect of each test substance on tumor weight of human pancreatic cancer Mia PaCa-2 nude mouse xenograft model animal. DETAILED DESCRIPTION

[0192] The application will be further described in the following by way of examples without limiting the application to the examples described. The experimental methods in the following examples, unless otherwise specified, are carried out according to the conventional methods and conditions, or according to the instructions of the commercial products.

[0193] Unless otherwise specified, the equipment and detection methods used in the present application are as follows:

[0194]

[0195]

[0196] Example 1: Preparation of the compound shown in formula (I)

[0197] First step, synthesis of compound B

[0198]

[0199] Reaction kettle R1 was added with ethanol (5.33 kg) and stirred. A (1.35 kg) was added. While maintaining the temperature at 5-15 °C, triethylamine (1.26 kg) was added dropwise. While maintaining the temperature at 5-15 °C, A2 (1.13 kg) was added dropwise. After the dropwise addition, the system was stirred at 5-15 °C for 0.5 h. The system was warmed to 30-35 °C and stirred for reaction. The reaction was detected by HPLC and the reaction endpoint was reached. R1 was concentrated to remove ethanol. Water (5.40 kg) and ethyl acetate (4.86 kg) were added to R1 and stirred for 10 min. The system was allowed to stand and the upper organic phase was temporarily stored in R1 and the lower aqueous phase was transferred to R2. Ethyl acetate (4.86 kg) was added to R2 and stirred for 10 min. The system was allowed to stand and the upper organic phase was transferred to R1 and the lower aqueous phase was temporarily stored in R2. Ethyl acetate (4.86 kg) was added to R2 and stirred for 10 min. The system was allowed to stand and the upper organic phase was transferred to R1 and the lower aqueous phase was discarded. Water (5.40 kg) was added to R1 and stirred for 10 min. The system was allowed to stand and the upper organic phase was collected and the lower aqueous phase was discarded. The organic phase was concentrated to dryness to obtain 1.89 kg of yellow oil.

[0200] 1 H NMR (400 MHz, CDCl3) δ 6.15 (s, 1H), 5.46 (s, 1H), 4.49 (t, J = 5.2 Hz, 1H), 3.59-3.45 (m, 2H), 3.42 (s, 6H), 2.32 (s, 3H).

[0201] Second step, synthesis of compound C

[0202]

[0203] Into reactor R1, add glacial acetic acid (6.51 kg) and start stirring. Add B (1.24 kg). Add NIS (1.33 kg). Warm the reaction to 30-35 °C and react for 4-6 h. HPLC check, reaction is complete. Cool the system to 5-10 °C and slowly add water (11.20 kg) to the system. After the addition is complete, stir for 16 h at 5-15 °C. Filter, and rinse the filter cake with water (2.50 kg). Collect the solid and dry under vacuum at 50 °C for 24 h. Yield 1.61 kg of crude solid, weight yield: 130.1%. Directly charge into the next reaction. 1 H NMR (400 MHz, CDC13) δ 5.87 (s, 1H), 4.52 (t, J = 5.2 Hz, 1H), 3.67 (t, J = 5.4 Hz, 2H), 3.48 (s, 6H), 2.59 (s, 3H).

[0204] Third step, synthesis of compound D

[0205]

[0206] Into reactor R1, add glacial acetic acid (6.72 kg) and start stirring. Add C (0.80 kg). Add concentrated hydrochloric acid (0.47 kg) dropwise. After the addition is complete, warm the reaction to 100-105 °C and react for 1-2 h. HPLC check, reaction is complete. Cool the system rapidly to 10-20 °C and filter. Rinse the filter cake with ethanol (0.63 kg). Collect the filter cake and transfer to R1. Into R1, add water (5.60 kg) and start stirring. Dissolve sodium bicarbonate (0.21 kg) in water (2.40 kg) and prepare a saturated solution. Add the solution to R1 dropwise. Stir for 1 h after the addition is complete. Filter and rinse the filter cake with water (0.80 kg). Collect the solid and dry under vacuum at 50 °C for 12 h. Constant weight gives 0.34 kg of solid, weight yield: 41.8%. 1 H NMR (400 MHz, DMSO) δ 10.52 (s, 1H), 6.85 (s, 1H), 5.88 (t, J = 5.9 Hz, 1H), 3.83 (dd, J = 15.4, 7.2 Hz, 1H), 3.42 (dd, J = 15.5, 1.8 Hz, 1H), 2.21 (s, 3H).

[0207] Fourth step, synthesis of compound E

[0208]

[0209] Into reactor Rl, add D (1.00 kg). Add phosphorous oxychloride (8.38 kg) and start stirring. Heat to 90-95°C and react for 3-4 hours. While maintaining the temperature at 90-95°C, add DIPEA (2.65 kg) dropwise. After the addition is complete, maintain the temperature at 90-95°C and stir for 2 hours. HPLC analysis shows that the reaction is complete. Concentrate the reaction mixture to remove as much of the phosphorous oxychloride as possible, and obtain a black oil. Add toluene (5.00 kg) to the concentrate and concentrate to a minimum amount of liquid. Dilute the concentrate with dichloromethane (13.30 kg). Into reactor R2, add water (20.00 kg) and start stirring. Slowly pour the dichloromethane solution from Rl into R2 to quench the reaction, maintaining the temperature at 30°C or less. After the quenching is complete, allow the mixture to separate into layers and remove the lower layer. Add dichloromethane (13.30 kg) to reactor R2 and stir for 20 minutes. Allow the mixture to separate into layers and remove the lower layer. Set aside the aqueous layer. Combine the dichloromethane layers and add 2N hydrochloric acid (5.00 kg) and stir for 20 minutes. Allow the mixture to separate into layers and combine the aqueous layers. Set aside the dichloromethane layer. Add 30% sodium hydroxide to the combined aqueous layers to adjust the pH to 6. Add ethyl acetate (36.00 kg) to reactor R2 and stir for 30 minutes. Stop stirring and filter the mixture through diatomaceous earth. Separate the layers and discard the lower aqueous layer. Set aside the upper organic layer in reactor R2. Add 5% sodium carbonate solution (5.00 kg) to reactor R2 and stir for 10 minutes. Allow the mixture to separate into layers and discard the lower aqueous layer. Set aside the upper organic layer in reactor R2. Add 5% sodium carbonate solution (5.00 kg) to reactor R2 and stir for 10 minutes. Allow the mixture to separate into layers and discard the lower aqueous layer. Set aside the upper organic layer in reactor R2. Add water (10.00 kg, 10.00X) to reactor R2 and stir for 10 minutes. Allow the mixture to separate into layers and discard the lower aqueous layer. Collect the upper organic layer. Concentrate the organic layer to dryness and obtain a yellow solid. Dry the solid at 50°C under vacuum for 24 hours and sieve the product. Obtain 0.51 kg of product with a yield of 51.0%.

[0210] Fifth Step, Synthesis of Compound F

[0211]

[0212] Into a reaction flask, add E (10.00 g, 1.00 eq), E2 (9.66 g, 1.03 eq), acetonitrile (150 ml, 15V), and stir at room temperature. Add DIPEA (22.02 g, 5.00 eq) and heat to 30°C. Stir for 4 hours and take a sample to confirm that the reaction is complete. Concentrate the system at 35°C to 2-3V. Add water (450 ml, 45V) and stir overnight. Filter the mixture to obtain a light yellow solid. Dry the product at 49°C to obtain the product (HPLC: 97.4%, yield: 99.8%). 1H NMR (400 MHz, DMSO) δ 7.79 (d, J = 1.3 Hz, 1H), 7.57 (d, J = 1.3 Hz, 1H), 7.51 (d, J = 6.9 Hz, 1H), 7.34 - 7.23 (m, 3H), 4.28 (s, 1H), 3.74 (t, J = 13.5 Hz, 2H), 3.25 - 3.05 (m, 3H), 2.90 (d, J = 16.0 Hz, 1H), 2.58 (s, 3H), 1.95 (dd, J = 13.2, 7.0 Hz, 2H), 1.57 (s, 2H), MS: m / z = 460.2 [M+l].

[0213] Step six, synthesis of compound G

[0214]

[0215] To the reaction bottle, F (5.0 g, 1.0 eq), MeOH (30 ml, 10V), start stirring, make the system clear, drop DIPEA (2.21 g, 1.5 eq), drop BOC2O (2.85 g, 1.2 eq) methanol solution (20 ml), after drop, keep 0-10 ℃ for 10 min, then warm to room temperature, the system is dried, add water (50 ml), EA extraction (50 ml*2), water washing (50 ml*2), NaCl aqueous solution washing (50 ml), concentrated to dryness, get 4.88 g product. 1 H NMR (400 MHz, DMSO) δ 7.79 (d, J = 1.3 Hz, 1H), 7.57 (d, J = 1.3 Hz, 1H), 7.51 (d, J = 6.9 Hz, 1H), 7.34 - 7.23 (m, 3H), 4.28 (s, 1H), 3.74 (t, J = 13.5 Hz, 2H), 3.25 - 3.05 (m, 3H), 2.90 (d, J = 16.0 Hz, 1H), 2.58 (s, 3H), 1.95 (dd, J = 13.2, 7.0 Hz, 2H), 1.57 (s, 2H), MS: m / z = 460.2 [M+l].

[0216] Step seven, synthesis of compound H

[0217]

[0218] Into a 100ml flask, add CuI (35mg, 0.2eq), TMEDA (63mg, 0.6eq), dioxane (5ml, 10V), replace with nitrogen for 3 times, stir for 1h at room temperature, add G (500mg, 1.0eq), G2 (239mg, 1.5eq), Cs2CO3 (440mg, 1.5eq), replace with nitrogen for 3 times again, heat to 101℃, stir for 24h, take sample to determine the reaction, after the reaction is completed, add 10ml of ammonia, filter (add diatomite), wash the filter cake with ethyl acetate (5V), shake, separate, extract with ethyl acetate (50ml), combine the organic phase, wash with water, wash with brine, dry with anhydrous sodium sulfate, filter and spin dry, add ethyl acetate to dissolve completely, drop in normal heptane (EA: normal heptane = 3:1), slowly cool to room temperature, stir for 16h, filter, add the filter cake to a 100ml flask, add 10V of acetone, drop in 2.5V of water, stir at room temperature for 4h, filter, send for testing to determine purity, dry the filter cake at 50℃ to obtain a light yellow solid. 1 H NMR (400 MHz, DMSO) δ 7.75 (d, J = 1.2 Hz, 1H), 7.54 (d, J = 5.4 Hz, 1H), 7.47 (s, 1H), 7.30 - 7.14 (m, 5H), 6.30 (s, 2H), 5.70 (d, J = 5.4 Hz, 1H), 4.87 (d, J = 9.8 Hz, 1H), 3.93 - 3.70 (m, 2H), 3.37 (s, 2H), 3.11 (d, J = 15.8 Hz, 1H), 2.78 (d, J = 15.7 Hz, 1H), 2.46 (s, 3H), 1.93 - 1.76 (m, 2H), 1.69 (d, J = 13.4 Hz, 1H), 1.52 (d, J = 13.6 Hz, 1H), 1.41 (d, J = 11.7 Hz, 9H), MS: m / z = 592.2 [M+l].

[0219] Eighth step, synthesis of the compound represented by formula (I)

[0220]

[0221] Compound H (0.3g, 0.51mmol) was dissolved in DCM (10ml), drop in TFA (2ml) at room temperature, stir for 1h at room temperature. Concentrate, dilute with DCM, pour into saturated aqueous Na2CO3 solution, extract with DCM 3 times, spin dry at 30-40℃ to obtain a white solid (203mg). 1H NMR (400 MHz, DMSO) δ 7.74 (d, J = 1.1 Hz, 1H), 7.54 (d, J = 5.4 Hz, 1H), 7.48 (d, J = 1.2 Hz, 1H), 7.33 (d, J = 6.3 Hz, 1H), 7.29 - 7.08 (m, 4H), 6.31 (s, 2H), 5.71 (d, J = 5.4 Hz, 1H), 3.31 - 3.20 (m, 2H), 3.08 (d, J = 15.6 Hz, 1H), 2.66 (d, J = 15.6 Hz, 1H), 2.47 (s, 3H), 2.06 - 1.88 (m, 3H), 1.79 (s, 2H), 1.63 (d, J = 13.3 Hz, 1H), 1.25 (d, J = 13.3 Hz, 1H). MS: m / z = 492.2 [M+1]. The obtained white solid is the compound of formula (I), which is determined as the crystal form of the compound of formula (I) A by PXRD spectrum. The PXRD spectrum is shown in Figure 1 .

[0222] The DSC spectrum is shown in Figure 2 , and the results show that the compound of formula (I) A crystal form presents the melting endothermic (Onset: 180.54°C, Peak: 188.96°C, ΔH = 22.96 J / g), crystal transformation exothermic (Onset: 190.55°C, Peak: 191.68°C, ΔH = 10.84 J / g), and melting endothermic (Onset: 207.29°C, Peak: 215.23°C, ΔH = 73.75 J / g) phenomena in the range of 30°C to 300°C, which indicates that there is a more stable crystal form of the compound in thermodynamics. The TGA spectrum is shown in Figure 3 , which shows that the compound of formula (I) A crystal form begins to decompose at about 278±3°C, and there is almost no weight loss before the decomposition temperature, indicating that the compound is an anhydrous and solvent-free crystal form.

[0223] Example Two: Preparation of the compound of formula (I) B crystal form

[0224] 300.6 mg of the compound of formula (I) A crystal form was weighed into a 20 mL sample bottle, 10 ml of acetonitrile was added, and it was shaken in a shaking bed (25°C, 250 rpm) for 24 h. Filtration was performed, and the obtained solid was placed in a 50°C vacuum drying box (vacuum degree -0.1 MPa) for drying for 6 h to obtain a white solid, with a yield of 64.45%. The solid was dried in vacuum and then subjected to PXRD determination, and the spectrum is shown in Figure 4 . The DSC and TGA results show that the compound is an anhydrous and solvent-free crystal form. The DSC spectrum is shown in Figure 5 , and the TGA spectrum is shown in Figure 6 , and the H 1 NMR spectrum is shown in Figure 19 .

[0225] The compound B of formula (I) is a non-solvate, and the DSC result shows that it is the most thermodynamically stable system.

[0226] Example Three: Preparation of the compound A of formula (III) in a crystal form

[0227]

[0228] The compound A of formula (I) in a crystal form 299.9 mg was weighed into a 20 mL sample bottle, 6 mL of dioxane was added, and the temperature was raised to 50°C while being stirred magnetically. After the solution was clear, 600 μL of methyl methanesulfonate (1 mol / L) was slowly added dropwise, the reaction temperature was 50°C, and the reaction was carried out for 3 h. Subsequently, the temperature was lowered to room temperature at a rate of 10°C / h, and then placed in a 4°C refrigerator for 24 h. Filtration was carried out, the filter cake was washed with 2 mL of dioxane at 4°C, and the filtered sample was placed in a 50°C vacuum drying oven (vacuum degree -0.1 MPa) for drying for 24 h, to obtain a white solid, with a yield of 74.68%. The product was subjected to PXRD determination, and the spectrum is shown in Figure 7 . The DSC and TGA results show that the compound is an anhydrous and solvent-free crystal form, the DSC spectrum is shown in Figure 8 , and the TGA spectrum is shown in Figure 9 , the H 1 NMR spectrum is shown in Figure 20 .

[0229] Example Four: Preparation of the compound A of formula (IV) in a crystal form

[0230]

[0231] The compound A of formula (I) in a crystal form 300.6 mg was weighed into a 50 mL crystallizer, 27 mL of acetonitrile and 3 mL of purified water were added, the temperature was raised to 50°C, and stirring was carried out uniformly (200-300 rpm). 600 μL of a citric acid methanol solution (1 mol / L) was slowly added dropwise, the reaction temperature was 50°C, the stirring speed was 200-300 rpm, and the reaction was carried out for 2 h. The temperature was lowered to 5°C at a rate of 15°C / h, and the crystal was aged for 0.5 h. The product was collected, the filter cake was washed with 2 mL of acetonitrile at 4°C, and the filtered solid was placed in a 50°C vacuum drying oven (vacuum degree -0.1 MPa) for drying for 24 h, to obtain a white solid, with a yield of 52.40%. The product was subjected to PXRD determination, and the spectrum is shown in Figure 10 . The DSC and TGA results show that the compound is an anhydrous and solvent-free crystal form 11, the DSC spectrum is shown in Figure 11 , and the TGA spectrum is shown in Figure 12 , the H 1 NMR spectrum is shown in Figure 21 .

[0232] Example Five: Preparation of Compound A crystalline form of formula (V)

[0233]

[0234] Example Five: Preparation of Compound A crystalline form of formula (V) Figure 13 The DSC and TGA results show that the compound is anhydrous and solvent-free crystalline form, the DSC spectrum is shown in Figure 14 , and the TGA spectrum is shown in Figure 15 , and the H 1 NMR spectrum is shown in Figure 22 .

[0235] Example Six: Preparation of Compound A crystalline form of formula (VI)

[0236]

[0237] Example Five: Preparation of Compound A crystalline form of formula (V) Figure 16 The DSC and TGA results show that the compound is anhydrous and solvent-free crystalline form, the DSC spectrum is shown in Figure 17 , and the TGA spectrum is shown in Figure 18 , and the H 1 NMR spectrum is shown in Figure 23 .

[0238] Example Seven: Water adsorption and desorption experiment of the compound crystalline form

[0239] The dynamic water adsorption instrument (DVS) was used to investigate the adsorption and desorption experiments of the above compound crystal forms at 25℃ and 0-95% relative humidity range, so as to determine the moisture absorption performance of various crystal forms. The experimental results are shown in Table 6.

[0240] Table 6: Weight change of sample in 0-95% RH range

[0241] crystalline form hygroscopic weight gain crystalline form of compound A represented by formula (I) 0.26％ crystalline form of compound B represented by formula (I) 0.13％ crystalline form of compound A represented by formula (III) 1.37％ crystalline form of compound A represented by formula (IV) 3.25％ crystalline form of compound A represented by formula (V) 5.38％ crystalline form of compound A represented by formula (VI) 1.28％

[0242] From the detection results, the moisture absorption weight gain of the compound B represented by formula (I) is less than 0.2%, almost no moisture absorption; the moisture absorption weight gain of the compound represented by formula (III), the compound represented by formula (VI) and the compound A represented by formula (I) is less than 2% but not less than 0.2%, slightly moisture absorption; the moisture absorption weight gain of the compound A represented by formula (V) and the compound A represented by formula (IV) is less than 15% but not less than 2%, moisture absorption.

[0243] Example Eight: Compound Crystal Form Solubility Test

[0244] 10mg of the above compound crystal form was weighed into a 10mL sample bottle, 5mL of water, pH 2.0, 4.5, 6.8 buffer was added respectively, and after shaking at 25℃ for 24h, it was filtered, and the filtrate was determined by HPLC for solubility and pH value.

[0245] The chromatographic conditions are as follows:

[0246] Chromatographic column: Unitary C18 (5μm, 100A, 4.6×250mm)

[0247] Mobile phase: A phase is 0.1% methanol solution, B phase is acetonitrile, A:B=10:90 gradient table:

[0248] T (min) A(％) B(％) 0 95 5 10 5 95 13 95 5 15 95 5

[0249] Detection wavelength: 254nm

[0250] Column temperature: 40℃

[0251] Injection volume: 20μL

[0252] The solubility test results are shown in Table 7.

[0253] Table 7: Solubility of compound crystal form (25℃, mg / mL)

[0254]

[0255]

[0256] *The solubility of the salt is converted into the solubility of the free base

[0257] The solubility test results show that the solubility of the compound of formula (I) crystal form is obviously pH-dependent, and the solubility of the weakly basic drug increases as the pH value decreases. The solubility of the compound of formula (I) B crystal form in pH 2.0 buffer salt solution and pH 4.5 buffer salt solution is equivalent to that of the compound of formula (I) A crystal form, but the solubility in pH 6.8 buffer salt solution and deionized water is much smaller than that of the compound of formula (I) A crystal form. Compared with the compound of formula (I) stable B crystal form, the solubility of the compound in pH 6.8 buffer salt solution and deionized water can be significantly improved after salification; compared with the compound of formula (I) A crystal form, the solubility in deionized water is also significantly improved after salification, among which the compound of formula (III) A crystal form is the best, which is increased by 82 times, and the solubility in the remaining pH buffer salt solution is also equivalent.

[0258] Example Nine: Determination of the stability of the crystal form of the compound

[0259] High temperature test (T): the powder is placed in a suitable sealed glass bottle, and is placed at 60°C for 10 days, and samples are taken on the 5th day and the 10th day for solid PXRD test.

[0260] High humidity test (H): the powder is opened and placed in a constant temperature and humidity chamber, and is placed at 25°C, 90%±5%RH for 10 days, and samples are taken on the 5th day and the 10th day for solid PXRD test. The hygroscopic deliquescence performance is investigated.

[0261] Strong light irradiation test (L): the powder is opened and placed in a light stability chamber equipped with a daylight lamp, and is placed at an illuminance of 4500±500lx for 10 days, and samples are taken on the 5th day and the 10th day for solid PXRD test.

[0262] Accelerated test (A): the powder is opened and placed in a constant temperature and humidity chamber, and is placed at 40°C, 75%±5%RH for 10 days, and samples are taken on the 5th day and the 10th day for solid PXRD test.

[0263] The stability test results are shown in Table 8:

[0264] Table 8: Stability of the crystal form of each compound

[0265]

[0266] The stability results show that the compound of formula (I) A, B, the compound of formula (III) A crystal form, and the compound of formula (IV) A crystal form are stable under each experimental condition and do not undergo crystal form transformation; however, the compound of formula (V) A crystal form partially undergoes crystal form transformation under accelerated conditions, and the compound of formula (VI) A crystal form undergoes crystal form transformation under high humidity and accelerated conditions.

[0267] Example 10: Hetero-transplant tumor model experiment of compound A crystal form of formula (IV)

[0268] Experiment 1

[0269] Animals: Balb / c nude mice, female, body weight 17-19 g, provided by Zhejiang Vantoll Life Science and Technology Co., Ltd.; SPF level feeding, temperature 20-26℃, humidity 40-70%, free feeding, city tap water after filtration and high pressure sterilization for drinking. The mice were given adaptive feeding for no less than 7 days before the experiment.

[0270] Preparation of administration solution: the vehicle of the test substance RG001 (i.e. compound A crystal form of formula (IV)) is 1% HPMC. The administration solution of the test substance is prepared on site, stored at 2-8℃, and protected from light.

[0271] Human cancer cell line: human non-small cell lung cancer cell line NCI-H358 was provided by the Institute of Cell Biology, Chinese Academy of Sciences.

[0272] Culture medium: RPMI-1640 basal medium and fetal bovine serum (FBS) were purchased from GIBCO Company (Grand Island, NY, USA).

[0273] Tumor transplantation experiment and detection: NCI-H358 cells were cultured in RPMI-1640 medium containing 10% FBS, and the cells were placed in a 5% CO2 incubator for culture at 37℃. The logarithmic growth phase of NCI-H358 cells was collected, resuspended in RPMI-1640 basal medium after counting, and 1:1 Matrigel was added. The concentration of the cell suspension was adjusted to 1*10 8 mL, 0.1 mL of the cell suspension was inoculated subcutaneously on the right side of the back of the mouse under sterile conditions.

[0274] When the average volume of the tumor reached 120mm 3 left and right, the animals were randomly divided into groups, with 8 mice in each group. Day 0 was recorded on the day of grouping, and the administration was started according to the average body weight. The animal body weight and tumor size were measured twice a week during the experiment. Compound A crystal form of formula (IV) group was administered orally once a day or once a week, continuously for three weeks. The positive drug group was administered orally with RMC4550 once a day, continuously for three weeks.

[0275] In this study, the experimental data are expressed as MEAN ± SEM. The tumor growth curve was plotted with time point as X-axis and tumor volume as Y-axis; the animal body weight change curve was plotted with time point as X-axis and animal body weight as Y-axis. Two-tailed t-test was used for comparison between groups, P<0.05 was considered as significant difference, and P<0.01 was considered as extremely significant difference.

[0276] In this experiment, all groups of animals in good condition, no significant change in body weight, no animal deaths, drug administration period, animals were tolerated to each test.

[0277] The experimental results show that compared with the blank solvent control group, the tumor weight of each administration group is significantly reduced. The RG001 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 4 mg / kg group is orally administered every day, continuously administered for 21 days, and the tumor weight inhibition rate of each group on the 21st day is 66.75%, 66.50%, 88.67%, and 90.86%, respectively. The RG001 14 mg / kg group is administered once a week for a total of three times, and the tumor weight inhibition rate is 80.88%. The positive control RMC4550 15 mg / kg group has a tumor weight inhibition rate of 79.42%. The specific results are shown in Table 1. Figure 25 . Figure 25 ** corresponds to P < 0.05, ** corresponds to P < 0.01, and ** corresponds to P < 0.001.

[0278] Compared with the blank solvent control group, the tumor of each administration group is significantly inhibited.

[0279] The RG001 0.5 mg / kg, 1 mg / kg, 2 mg / kg, 4 mg / kg group is orally administered every day, continuously administered for 21 days, and the relative tumor inhibition rate T / C of each group on the 21st day is 33.33%, 31.58%, 12.4%, and 10.06%, respectively. The RG001 14 mg / kg group is administered once a week for a total of three times, and the T / C on the 21st day is 23.39%. The positive control RMC4550 15 mg / kg group is orally administered every day, continuously administered for 21 days, and the T / C on the 21st day is 20.70%. The specific experimental results are shown in Table 2. Figure 24 .

[0280] Experiment Two

[0281] Animals: SCID mice, female, body weight 15-18 g, animal source from Beijing Vital River Laboratory Animal Technology Co., Ltd.; SPF level feeding, temperature 20-26℃, humidity 40-70%, free feeding, city tap water after filtration and high pressure sterilization for drinking. The mice were given 7 days of adaptive feeding before the experiment.

[0282] Drug solution preparation: the solvent of the test substance RG001 (compound A crystal form shown in formula (IV)) is 1% HPMC. The test substance administration solution is prepared on site, stored at 2-8℃, and protected from light.

[0283] Human cancer cell line: human leukemia cell line MV-4-11 was provided by ATCC (American Type Culture Collection, USA).

[0284] Culture medium: IMDM basal medium and fetal bovine serum (FBS) were purchased from GIBCO (Grand Island, NY, USA).

[0285] Tumor transplantation and detection: MV-4-11 cells were cultured in IMDM medium containing 10% FBS, and the cells were placed in a 5% CO2 incubator at 37°C. The logarithmic growth phase MV-4-11 cells were collected, counted and resuspended in IMDM basal medium, 1:1 Matrigel was added, and the cell suspension concentration was adjusted to 5*10 7 mL, 0.1 mL of cell suspension was inoculated subcutaneously on the right side of the back of the mouse under sterile conditions, and the inoculation concentration was 5*10 6 / 0.1 mL / mouse.

[0286] When the average tumor volume reached 120 mm 3 left and right, the animals were randomly divided into groups, with 8 mice in each group. Day 0 was recorded on the day of grouping, and the drug administration was started according to the average body weight. During the experiment, the animal body weight and tumor size were measured twice a week. The experimental group was orally administered once a day or once a week, and the administration was continued for three weeks. The positive drug group was orally administered RMC4550 once a day, and the administration was continued for three weeks.

[0287] In this study, the experimental data were expressed as MEAN ± SEM. The tumor growth curve was plotted with time point as X axis and tumor volume as Y axis; the animal body weight change curve was plotted with time point as X axis and animal body weight as Y axis. Two-tailed t-test was used for comparison between groups, P<0.05 was significant difference, and P<0.01 was extremely significant difference.

[0288] In this experiment, all the animals were in good condition, and there was no significant change in body weight, and no animal died. During the administration period, the animals could tolerate each test substance.

[0289] The experimental results showed that compared with the blank solvent control group, the tumor weight of each administration group was significantly reduced. The RG001 1 mg / kg, 2 mg / kg, and 4 mg / kg groups were orally administered every day, and the administration was continued for 21 days. The inhibition rates of tumor weight of 1 mg / kg, 2 mg / kg, and 4 mg / kg groups were 67.13%, 93.72%, respectively; the tumor of the 4 mg / kg group completely regressed; the RG001 14 mg / kg group was administered once a week, and the administration was continued for three times, and the inhibition rate of tumor weight was 88.46%. The positive control RMC4550 15 mg / kg group was administered every day, and the administration was continued for 21 days, and the inhibition rate of tumor weight was 88.37%. The specific results are shown in Table 1. Figure 27 . Figure 27 ** corresponds to P<0.05, ** corresponds to P<0.01, and ** corresponds to P<0.001.

[0290] Compared with the blank solvent control group, the tumors of each administration group were significantly inhibited. The 1 mg / kg, 2 mg / kg, and 4 mg / kg groups of RG001 were orally administered daily for 21 days. The relative tumor growth rates T / C of the 1 mg / kg and 2 mg / kg dose groups were 33.22% and 6.81%, respectively. The tumors of the 4 mg / kg group completely regressed. The 14 mg / kg group of RG001 was administered once a week for a total of three times. The T / C on day 21 was 12.95%. The positive control RMC4550 15 mg / kg group was orally administered daily for 21 days. The T / C on day 21 was 8.94%. The specific results are shown in Table 1. Figure 26 .

[0291] Experiment Three

[0292] Animals: Balb / c nude mice, female, weighing 17-19 g, provided by Zhejiang Vantoll Life Experimental Animal Technology Co., Ltd.; SPF level feeding, temperature 20-26°C, humidity 40-70%, free feeding, city tap water filtered and autoclaved for drinking. The mice were given adaptive feeding for no less than 7 days before the experiment.

[0293] Preparation of the administration solution: the solvent of the test substance RG001 (compound A in formula (IV) in the form of a crystal) is 1% HPMC. The administration solution of the test substance is prepared on site, stored at 2-8°C, and protected from light.

[0294] Human cancer cell line: human pancreatic cancer cell line Mia PaCa-2 was provided by ATCC (American Type Culture Collection, USA).

[0295] Culture medium: DMEM basic medium and fetal bovine serum (FBS) were purchased from GIBCO (Grand Island, NY USA).

[0296] Tumor transplantation experiment and detection: Mia PaCa-2 cells were cultured in DMEM medium containing 10% FBS. The cells were placed in a 5% CO2 incubator at 37°C. The NCI-H358 cells in the logarithmic growth phase were collected, counted, and resuspended in DMEM basic medium. Matrigel was added at a ratio of 1:1, and the cell suspension concentration was adjusted. Under sterile conditions, 0.1 mL of cell suspension was inoculated subcutaneously into the right dorsal skin of the mice.

[0297] When the average tumor volume reached 120 mm 3When the tumor volume reached 100mm3, the animals were randomly divided into groups by randomized block method, 8 mice in each group. The day of grouping was recorded as Day 0, and the animals were administered according to the average body weight, with a 28-day experimental period. The body weight and tumor size of the animals were measured twice a week during the experiment. Compound A in the form of formula (IV) was administered orally once a day or once a week for four consecutive weeks. The positive drug group was administered RMC4550 orally once a day for four consecutive weeks.

[0298] In this study, the experimental data are expressed as MEAN ± SEM. The tumor growth curve was plotted with the time point as the X-axis and the tumor volume as the Y-axis. The animal weight change curve was plotted with the time point as the X-axis and the animal weight as the Y-axis. Two-tailed t-test was used for comparison between groups, with P < 0.05 as significant difference and P < 0.01 as extremely significant difference.

[0299] In this experiment, all animals were in good condition, with no significant changes in body weight, and no animal deaths occurred. During the administration period, the animals were able to tolerate each test substance.

[0300] The experimental results showed that compared with the blank solvent control group, the tumor weight of each administration group was significantly reduced. The 1 mg / kg, 2 mg / kg, and 4 mg / kg groups of RG001 were administered orally every day for 28 consecutive days, and the inhibition rates of tumor weight in each dose group were 76.36%, 79.08%, and 81.66%, respectively. The 14 mg / kg group of RG001 was administered once a week for a total of four times, and the inhibition rate of tumor weight was 69.16%. The positive control RMC4550 15 mg / kg group was administered daily for 28 days, and the inhibition rate of tumor weight was 78.53%. The specific results are shown in Table 1. Figure 29 . Figure 29 ** corresponds to P < 0.05, ** corresponds to P < 0.01, and *** corresponds to P < 0.001.

[0301] Compared with the blank solvent control group, the tumor of each administration group was significantly inhibited. The 1 mg / kg, 2 mg / kg, and 4 mg / kg groups of RG001 were administered orally every day for 28 consecutive days, and the relative tumor growth rates T / C of each dose group were 27.61%, 20.55%, and 16.75%, respectively. The 14 mg / kg group of RG001 was administered once a week for a total of four times, and the T / C on the 28th day was 30.77%. The positive control RMC4550 15 mg / kg group was administered orally every day for 28 consecutive days, and the T / C on the 28th day was 20.02%. The specific results are shown in Table 2. Figure 28 .

[0302] Example XI: Pharmacokinetic experiment of compound A in the form of formula (I), compound A in the form of formula (III), and compound A in the form of formula (IV)

[0303] Drug and Reagent: (S)-1'-(8-((((2-amino-3-chloropyridin-4-yl)thio)-7-methylimidazo[1,2-c]pyrimidin-5-yl)-1,3-dihydrospiro[indene-2,4'-piperidin]-1-amine A crystal form (i.e. crystal form A of the compound of formula (I), crystal form A of the compound of formula (III), crystal form A of the compound of formula (IV) used in this study were ground into fine particles. The content (purity) of the material was not less than 95.0%.

[0304] Test Animals: SPF grade SD rats were divided into the compound of formula (I) group and each salt type of the compound of formula (I) group, each group including 3 male rats.

[0305] Drug Preparation: Preparation on the day of administration.

[0306] Firstly, solvent preparation: the required amount of deionized water was measured into a suitable container, and the required amount of HPMC was weighed and added and stirred to mix until uniform, to obtain a colorless clear 1% HPMC solution.

[0307] The required amount of compound was weighed into a suitable container, and the appropriate amount of 1% HPMC solution was added, stirred to uniform, and then 1% HPMC solution was added to the specified volume, stirred to uniform, to obtain a white suspension. The final concentration of each compound was 0.5 mg / mL.

[0308] Dosing and sample collection: each suspension was administered to SD rats at a dose volume of 10 mL / kg, with a dose of 5 mg / kg administered orally. The animals in the oral administration group were fasted overnight (10-16 hours) before administration, and fed 4 hours after administration. Blood was collected from the jugular vein before administration (0 h) and 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h after administration, with each sample collecting about 0.20 mL, K2EDTA anticoagulation, and placed on ice after collection, and centrifuged to separate plasma (centrifugation conditions: 6800g, 6 minutes, 2-8℃) within 1 hour. The plasma samples were stored at -80℃ for analysis.

[0309] The blood samples were collected in pre-anticoagulation tubes with EDTA-K. The plasma in the samples was separated by centrifugation at 4000 revolutions per minute for 10 minutes at 4℃. The plasma samples were collected and stored at -80℃ for analysis. The samples were analyzed by TQ5500 LC / MS combined with HPLC. The chromatographic column used in the liquid chromatography conditions was ACQUITY UPLC HSS T3 1.8 um (2.1*50mm) as the stationary phase, and 0.1% formic acid acetonitrile solution as the mobile phase. The specific experimental results are shown in Table 9:

[0310] Table 9

[0311]

[0312] The experimental results show that the compound A crystalline form of formula (IV) and the compound A crystalline form of formula (III) both exhibit more excellent metabolic properties than the compound A crystalline form of formula (I), and the exposure and maximum blood concentration are both greatly improved.

[0313] Example 12: Preparation method of tablets

[0314] The preparation formula of the tablets is shown in Table 10:

[0315] Table 10

[0316]

[0317] 1 Herein, the API specifically refers to the compound A crystalline form of formula (IV).

[0318] The preparation process of the tablets is as follows:

[0319] 1) Pretreatment of raw and auxiliary materials

[0320] The API and the remaining auxiliary materials are sieved through a 40-mesh screen to prevent caking.

[0321] 2) Blending

[0322] According to the batch prescription material information, the raw and auxiliary materials are weighed for standby.

[0323] 3) Sieving and premixing

[0324] The 0.25 mg and 2 mg specifications are mixed at the same time:

[0325] Most of the colloidal silicon dioxide (4 / 5) and part of the microcrystalline cellulose (3 / 5) are mixed at a speed of 200 rpm to 300 rpm, sieved through a 1.0 mm round hole screen, and passed through a powder crushing granulator once, serving as mixture one.

[0326] The compound A crystalline form of formula (IV) and an equal volume of colloidal silicon dioxide are mixed at a speed of 200 rpm to 300 rpm, sieved through a 1.0 mm round hole screen, and passed through a powder crushing granulator once, and then mixed with the remaining colloidal silicon dioxide at a speed of 200 rpm to 300 rpm, sieved through a 1.0 mm round hole screen, and passed through a powder crushing granulator once, serving as premixing mixture two.

[0327] The mixture one, mixture two, cross-linked polyvinylpyrrolidone, and the remaining microcrystalline cellulose are transferred to the hopper mixer, and the speed is set to 15 rpm, and mixed for 30 min.

[0328] 4) Total mixing

[0329] The prescribed amount of magnesium stearate was weighed and added to the premix, and the rotation speed was set to 15 rpm and mixed for 5 min. After the total mixing was completed, 10 points were measured for the mixing uniformity of the material in the upper, middle and lower layers of the hopper, with 5, 4 and 1 points in each layer.

[0330] 5) Tabletting

[0331] The mixing uniformity of the total mixed material was 95.0-105.0%, which met the intermediate quality standard, and the theoretical tablet weight was the labeled tablet weight.

[0332] Tabletting for 0.25 mg specifications:

[0333] A ZP10A rotary tablet machine was used for tabletting, with a 5.5 mm circular shallow concave punch, the rotation speed of the turntable was set to 15-25 rpm, the feed speed was 10-15 rpm, the tablet weight difference was required to be ±7%, and the tablet hardness was controlled to be 30-60 N.

[0334] Tabletting for 2 mg specifications:

[0335] A ZP10A rotary tablet machine was used for tabletting, with an 11 mm circular shallow concave punch, the rotation speed of the turntable was set to 15-25 rpm, the feed speed was 15-20 rpm, the tablet weight difference was required to be ±5%, and the tablet hardness was controlled to be 70-100 N.

[0336] 6) Packaging

[0337] The packaging material was an oral solid drug-use high-density polyethylene bottle and an oral solid drug-use polypropylene-low-density polyethylene child safety moisture-proof combined bottle cap.

[0338] Those skilled in the art will appreciate that, although specific embodiments of the application are described herein for illustrative purposes, various modifications can be made without departing from the spirit and scope of the application. Therefore, the specific embodiments and examples of the application should not be considered limiting the scope of the application. The application is limited only by the claims. All documents cited in this application are incorporated by reference in their entirety.

Claims

1. A crystal form, which is Form A of a compound of formula (IV) : ； wherein The X-ray powder diffraction pattern of the Form A of the compound of formula (IV) has characteristic diffraction peaks at the following 2Θ angles: 9.5±0.2°, 11.5±0.2°, 11.8±0.2°, 13.5±0.2°, 14.1±0.2°, 16.3±0.2°, 18.0±0.2°, 20.7±0.2° and 25.1±0.2°.

2. The crystal form as described in claim 1, characterized in that, The X-ray powder diffraction pattern of the Form A of the compound of formula (IV) has characteristic diffraction peaks at the following 2Θ angles: 9.5±0.2°, 10.3±0.2°, 11.5±0.2°, 11.8±0.2°, 12.2±0.2°, 12.9±0.2°, 13.5±0.2°, 14.1±0.2°, 15.7±0.2°, 16.3±0.2°, 17.0±0.2°, 18.0±0.2°, 18.6±0.2°, 20.7±0.2°, 21.1±0.2°, 22.2±0.2°, 23.2±0.2°, 23.8±0.2°, 24.5±0.2°, 24.8±0.2°, 25.1±0.2°, 26.2±0.2°, 28.7±0.2°, 29.4±0.2°, 29.7±0.2°, 30.5±0.2°, 31.8±0.2°, 32.3±0.2°, 33.9±0.2°, 34.6±0.2°, 35.4±0.2°, 36.5±0.2° and 40.4±0.2°. and / or, the differential scanning calorimetry (DSC) curve of the Form A of the compound of formula (IV) has endothermic peaks at 90.14℃, 162.81℃ and 189.67℃; and / or, the thermogravimetric analysis (TGA) curve of the Form A of the compound of formula (IV) shows a weight loss of 3.64% from room temperature to 115℃.

3. The crystal form as described in claim 2, characterized in that, The X-ray powder diffraction pattern of the Form A of the compound of formula (IV) has characteristic diffraction peaks at the following 2Θ angles: 9.5±0.2°, 11.5±0.2°, 11.8±0.2°, 13.5±0.2°, 14.1±0.2°, 16.3±0.2°, 18.0±0.2°, 20.7±0.2° and 25.1±0.2°. Table 3: ； and / or, the differential scanning calorimetry (DSC) curve of the Form A of the compound of formula (IV) has endothermic peaks at 90.14℃, 162.81℃ and 189.67℃; and / or, the thermogravimetric analysis (TGA) curve of the Form A of the compound of formula (IV) shows a weight loss of 3.64% from room temperature to 115℃.

4. The crystalline form of claim 3, characterized by The X-ray powder diffraction pattern of the Form A of the compound of formula (IV) has characteristic diffraction peaks at the following 2Θ angles: 9.5±0.2°, 11.5±0.2°, 11.8±0.2°, 13.5±0.2°, 14.1±0.2°, 16.3±0.2°, 18.0±0.2°, 20.7±0.2° and 25.1±0.2°.

5. A preparation method of the crystal form of claim 1, characterized in that: The preparation method of the Form A of the compound of formula (IV) comprises the following steps: mixing the Form A of the compound of formula (I) with acetonitrile and purified water, and then reacting with a methanol citric acid solution to crystallize to obtain the Form A of the compound of formula (IV). ； The X-ray powder diffraction pattern of the Form A of the compound of formula (I) is substantially as shown in Figure 1. The mass-volume ratio of the compound A crystalline form shown in formula (I) to the acetonitrile is 5-25 mg / mL; The mass-volume ratio of the compound A crystalline form shown in formula (I) to the purified water is 30-170 mg / mL; The operation of the reaction crystallization comprises the following steps: after the compound A crystalline form shown in formula (I) is mixed with acetonitrile and purified water and reacted with the citric acid methanol solution, cooling is performed, and the solid in the mixture after the reaction is separated out; The reaction is performed at a temperature of 50 ℃; the cooling operation comprises cooling to 5 ℃ at a cooling rate of 15 ℃ / h.

6. The production method according to claim 5, wherein In the preparation method of the compound A crystalline form shown in formula (IV), the mixing operation is stirring; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the mass-volume ratio of the compound A crystalline form shown in formula (I) to the citric acid methanol solution is 200-900 mg / mL; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the concentration of the citric acid methanol solution is 0.5-2 mol / L; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the solid is further washed after being separated out; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the solid is further dried after being separated out.

7. The production method according to claim 6, wherein In the preparation method of the compound A crystalline form shown in formula (IV), the mixing operation is stirring; the stirring is stirring while being warmed to 50 ℃; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the mass-volume ratio of the compound A crystalline form shown in formula (I) to the acetonitrile is 11.1 mg / mL; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the mass-volume ratio of the compound A crystalline form shown in formula (I) to the purified water is 100 mg / mL; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the mass-volume ratio of the compound A crystalline form shown in formula (I) to the citric acid methanol solution is 500 mg / mL; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the concentration of the citric acid methanol solution is 1 mol / L; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the reaction is performed at a temperature of 50 ℃; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the cooling operation comprises cooling to 5 ℃ at a cooling rate of 15 ℃ / h and then crystallizing for 0.5 h; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the solid is further washed after being separated out; the washing is washing with an acetonitrile solution; And / or, in the preparation method of the compound A crystalline form shown in formula (IV), the solid is further dried after being separated out; the drying is drying at a temperature of 50 ℃.

8. The production method according to claim 7, wherein The mixing operation is stirring; the stirring is stirring while heating to 50 ℃; And / or, in the preparation method of the compound A crystal form shown in formula (IV), the reaction is stirred at a temperature of 50 ℃ for 2 h; And / or, in the preparation method of the compound A crystal form shown in formula (IV), the solid is further washed after being separated out; the washing is washing with 4 ℃ acetonitrile solution; And / or, in the preparation method of the compound A crystal form shown in formula (IV), the solid is further dried after being separated out; the drying is drying at 50 ℃ and a vacuum degree of-0.1 M.

9. The production method according to claim 8, wherein In the preparation method of the compound A crystal form shown in formula (IV), the solid is further washed after being separated out; the mass / volume ratio of the compound A crystal form shown in formula (I) to acetonitrile solution is 150 mg / mL; And / or, in the preparation method of the compound A crystal form shown in formula (IV), the solid is further dried after being separated out; the drying is drying at 50 ℃ and a vacuum degree of-0.1 M for 24 h.

10. The production method according to claim 9, wherein In the preparation method of the compound A crystal form shown in formula (IV), the solid is further dried after being separated out; the drying is drying in a vacuum drying oven at 50 ℃ and a vacuum degree of-0.1 M for 24 h.

11. A pharmaceutical composition comprising: (1) a therapeutically effective amount of the crystal form according to any one of claims 1-4, and (2) a pharmaceutically acceptable excipient, adjuvant or carrier.

12. The pharmaceutical composition of claim 11, wherein, The pharmaceutical composition is for oral administration.

13. The pharmaceutical composition of claim 12, wherein The pharmaceutical composition is for preparing tablets or capsules.

14. The pharmaceutical composition of claim 13, wherein, The pharmaceutical composition contains 0.2%-10% by weight of the crystal form according to any one of claims 1-4.

15. Use of the crystal form according to any one of claims 1-4 or the pharmaceutical composition according to claim 11 in the preparation of a drug for treating, preventing, delaying or hindering the occurrence or progression of a disease associated with SHP2 protein activity or expression.

16. The use according to claim 15, wherein the compound is ###0009### The drug is a drug for treating a disease associated with SHP2 protein activity or expression.

17. The use according to claim 16, wherein The disease is a tumor.

18. The use of claim 17, wherein, The tumor is a tumor caused by abnormal Ras-Raf-ERK or PD1 / L1 signaling pathway.

19. The use according to claim 18, wherein the compound is ###0007### The tumor is esophageal cancer, lung cancer, colorectal cancer, pancreatic cancer, leukemia or gastric cancer.

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