Aryl phosphine oxide derivatives, acid salts or crystal forms thereof, and methods of making and using the same

Abstract

The present application relates to a kind of aryl phosphine oxide derivatives, acid salt or its crystal form and its preparation method and application, free base or salt of aryl phosphine oxide derivative and its crystal form, and the present application also relates to its preparation method and application.It is specifically related to a kind of crystal form of compound shown in general formula (I), salt and its crystal form and its preparation method, in addition, it is also related to the pharmaceutical composition containing therapeutically effective amount of the salt or crystal form of the compound and its use as EGFR inhibitor in the treatment of cancer related diseases.

Description

Technical Field

[0001] This invention belongs to the field of drug synthesis, specifically relating to the crystal form of an aryl phosphorus oxide derivative free base, its preparation method, and its application. Background Technology

[0002] EGFR (Epidermal Growth Factor Receptor) is a member of the ErbB family of transmembrane receptor tyrosine kinases, activated by binding to its ligands epidermal growth factor (EGF) or transforming growth factor α (TGFα). Activated EGFR forms homodimers on the cell membrane or heterodimers with other receptors in the family (such as ErbB-2, ErbB-3, or ErbB-4), leading to phosphorylation of key intracellular tyrosine residues of EGFR. This activates downstream signaling pathways, playing a crucial role in cell proliferation, survival, and anti-apoptosis. Activating mutations, overexpression, or gene amplification of EGFR can lead to excessive activation, promoting cell transformation into tumor cells and playing a vital role in tumor cell proliferation, invasion, metastasis, and angiogenesis. Therefore, EGFR is an important target for the development of anticancer drugs, particularly for lung cancer treatment.

[0003] First-generation EGFR small molecule inhibitors, including gefitinib (Iressa) and erlotinib (Tarceva), have shown good efficacy in the treatment of lung cancer and have been used as first-line drugs for the treatment of non-small cell lung cancer (NSCLC) with EGFR activating mutations (including L858R and delE746_A750). However, after 10-12 months of treatment with first-generation small molecule EGFR inhibitors, almost all NSCLC patients develop resistance to these inhibitors. More than half of these resistance mechanisms are due to secondary mutations in the EGFR gate gene residue T790M.

[0004] Osimertinib (AZD9291) is a third-generation EGFR TKI inhibitor with a high response rate and good therapeutic effect against EGFR T790M resistance. It received accelerated approval from the FDA in November 2015 and has proven effective in treating advanced non-small cell lung cancer (NSCLC) patients with EGFR T790M resistance mutations. Despite the significant success of osimertinib in treating EGFR T790M-mutant NSCLC, resistance inevitably develops after 9–14 months of treatment. Studies have shown that resistance in up to 20–40% of patients is due to the EGFR C797S mutation. The EGFR C797S mutation changes the cysteine ​​residue at position 797 to serine, preventing osimertinib from forming a covalent bond with the EGFR protein, thus leading to resistance. Currently, there are no effective inhibitors targeting the EGFR C797S resistance mutation. Therefore, there is an urgent need to develop novel, highly active EGFR inhibitors to address the drug resistance problem caused by EGFR C797S mutations.

[0005] Novartis reported EAI0450, a compound targeting EGFR C797S resistance. This EGFR allosteric inhibitor, when combined with EGFR monoclonal antibodies such as cetuximab, showed good antitumor efficacy in a mouse model with the L858R / T790M / C797S mutation. However, this compound was ineffective as a monotherapy and could not inhibit the C797S resistance mutation containing deIE746_A750, thus failing to enter clinical trials. In 2017, Ken Uchibori et al. reported that the combination of Brigatinib (AP26113) and EGFR monoclonal antibodies (such as cetuximab) could overcome resistance to third-generation EGFR inhibitors caused by the C797S mutation, showing good antitumor efficacy in a PC9 (EGFR-C797S / T790M / de119) mouse model. However, Brigatinib also faced the challenge of low in vitro activity as a monotherapy and no significant in vivo antitumor activity, and similarly, no further clinical studies were conducted.

[0006] Lung cancer is a major disease threatening human health, and its mortality rate ranks first among all malignant tumors. In my country, the incidence of lung cancer is rising year by year, with approximately 700,000 new cases annually. About 35% of all NSCLC cases in my country have EGFR activating mutations. While first- or third-generation EGFR inhibitors can achieve good therapeutic effects, new drug-resistant mutations often develop later. Therefore, developing a new generation of anti-drug-resistant EGFR inhibitors has enormous clinical and market value. Summary of the Invention

[0007] All contents contained in patents PCT / CN2020 / 097362 and PCT / CN2020 / 097369 are incorporated herein by reference.

[0008] The object of this invention is to provide a crystal form of the compound represented by general formula (I).

[0009]

[0010] in:

[0011] R1 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkylthio, haloalkoxy, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0012] R2 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkylthio, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0013] Alternatively, any two R2 atoms can be linked with the carbon atoms they are attached to to form a cycloalkyl or heterocyclic group;

[0014] Ring A is selected from cycloalkyl, heterocyclic, aryl, or heteroaryl groups;

[0015] R a Selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, hydroxyalkyl, cyanoalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -(CH2). n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0016] R aa and R bb Each is independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, or heteroaryl;

[0017] x is an integer between 0 and 4;

[0018] y is an integer between 0 and 4;

[0019] t is an integer between 0 and 1;

[0020] m is an integer between 0 and 2; and

[0021] n is an integer between 0 and 2.

[0022] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein ring A is selected from 3-12 membered heterocyclic groups; preferably 3-8 membered heterocyclic groups; more preferably 3-8 membered monocyclic heterocyclic groups or fused heterocyclic groups containing 1-2 N or O atoms; and further preferably the following groups:

[0023]

[0024] In a preferred embodiment of the present invention, the crystal form represented by general formula (I), wherein R a Selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Cyanoalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy group, -(CH2) n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0025] Preferred elements include hydrogen, deuterium, halogens, hydroxyl groups, and C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 1-3 Cyanoalkyl, C 1-3 Alkyl group, -(CH2) n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0026] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, hydroxyl, methyl, ethyl, isopropyl, -(CH2)2F, -CH2OH, -C(CH3)2OH, -CH2CN, -OCH2CH3, -CH2OCH3, -C(O)CH3, -S(O)2CH3, -N(CH3)2, -NCH3(CH2CH3) or

[0027] R aa and R bb Each is independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, haloalkoxy, C 2-6 alkenyl, C 2-6 alkynyl, cycloalkyl, heterocyclic, aryl or heteroaryl;

[0028] Preferred methyl, ethyl, or oxetine butyl groups;

[0029] x is an integer between 0 and 2.

[0030] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein R1 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Alkylthio;

[0031] Preferred elements: hydrogen, deuterium, halogens, C 1-6 Alkyl, C 1-3 Halogenated alkyl or C 1-3 Alkylthio;

[0032] More preferably, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, or methylthio;

[0033] The preferred compounds are chlorine, bromine, trifluoromethyl, or methylthio.

[0034] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein R2 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 alkenyl or C 2-6 alkynyl group;

[0035] Preferred elements: hydrogen, deuterium, halogens, C 1-6 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 2-4 alkynyl group;

[0036] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, methoxy, or ethynyl;

[0037] The preferred radicals are hydrogen, fluorine, methyl, ethyl, trifluoromethyl, methoxy, or ethynyl.

[0038] Alternatively, any two R2 atoms can be linked with the carbon atoms they are attached to form a 3-8 membered heterocyclic group; preferably a 5-6 membered heterocyclic group containing 1-2 N or O atoms; more preferably a tetrahydrofuran group;

[0039] y is an integer between 0 and 3.

[0040] In a more preferred embodiment of the present invention, the compound (6-((5-bromo-2-((2-methoxy-5-methyl-4-(4-(((3aR,6aS)-tetrahydro-1H-furano[3,4-c]pyrrolo-5(3H)-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation is provided;

[0041] (6-((5-chloro-2-((4-(4-(3-(dimethylamino)acetidin-1-yl)piperidin-1-yl)-5-ethyl-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation;

[0042] 2-(1-(1-(4-((5-bromo-4-((5-(dimethylphospho)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-yl)acetidin-3-yl)acetonitrile;

[0043] The crystalline form of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation.

[0044] The most preferred crystal forms are (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidized to crystal forms A, B, C, D, E, F, G, J, K, L, and M, wherein:

[0045] The X-ray powder diffraction pattern of crystal form A has a diffraction peak at 2θ of 17.7 ± 0.2°; or at 21.5 ± 0.2°; or at 23.9 ± 0.2°; or at 14.7 ± 0.2°; or at 7.7 ± 0.2°; or at 12.3 ± 0.2°; or at 15.2 ± 0.2°; or at 20.9 ± 0.2°. The diffraction peak is present at 2°; or at 24.4±0.2°; or at 18.9±0.2°; or at 30.8±0.2°; or at 23.5±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0046] As a specific example, the X-ray powder diffraction pattern of crystal form A contains at least one or more diffraction peaks located at 2θ of 17.7±0.2°, 21.5±0.2°, and 23.9±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, and 20.9±0.2°, preferably two, three, four, or five.

[0047] The X-ray powder diffraction pattern of crystal form A may optionally include one or more diffraction peaks located at 2θ of 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, 20.9±0.2°, 24.4±0.2°, and 18.9±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 of these peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0048] More preferably, the X-ray powder diffraction pattern of crystal form A includes one or more diffraction peaks located at 2θ of 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, 20.9±0.2°, 24.4±0.2°, 18.9±0.2°, 30.8±0.2°, 23.5±0.2°, 13.0±0.2°, 16.9±0.2°, and 17.3±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 locations.

[0049] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 1.

[0050] Table 1

[0051]

[0052]

[0053] The crystalline form A of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 1 As shown; its DSC spectrum is basically as follows Figure 2 As shown; its TGA spectrum is basically as follows. Figure 3 As shown.

[0054] In a further preferred embodiment of the present invention, the crystalline form B is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0055] The X-ray powder diffraction pattern of crystal form B shows a diffraction peak at 2θ = 22.9 ± 0.2°; or at 20.6 ± 0.2°; or at 17.9 ± 0.2°; or at 15.2 ± 0.2°; or at 19.4 ± 0.2°; or at 12.4 ± 0.2°; or at 26.0 ± 0.2°; or at 22.5 ± 0.2°. The diffraction peak is present at 0.2°; or at 8.1±0.2°; or at 15.7±0.2°; or at 21.3±0.2°; or at 14.6±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0056] The X-ray powder diffraction pattern of crystal form B contains at least one or more diffraction peaks located at 2θ of 22.9±0.2°, 20.6±0.2°, and 17.9±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, and 22.5±0.2°, preferably two, three, four, or five.

[0057] The X-ray powder diffraction pattern of crystal form B may optionally include one or more diffraction peaks located at 2θ of 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, 22.5±0.2°, 8.1±0.2°, and 15.7±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 of these peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0058] More preferably, the X-ray powder diffraction pattern of crystal form B includes one or more diffraction peaks located at 2θ of 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, 22.5±0.2°, 8.1±0.2°, 15.7±0.2°, 21.3±0.2°, 14.6±0.2°, 29.4±0.2°, 32.0±0.2°, and 32.9±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 locations.

[0059] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 2.

[0060] Table 2

[0061]

[0062]

[0063] The present invention relates to the crystalline form B of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation, the X-ray powder diffraction pattern of which is basically as follows. Figure 4 As shown; its DSC spectrum is basically as follows Figure 5 As shown.

[0064] In a further preferred embodiment of the present invention, the crystalline form C is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0065] The X-ray powder diffraction pattern of crystalline form C shows a diffraction peak at 2θ of 19.0 ± 0.2°; or at 26.5 ± 0.2°; or at 9.2 ± 0.2°; or at 25.3 ± 0.2°; or at 4.8 ± 0.2°; or at 14.1 ± 0.2°; or at 21.5 ± 0.2°; or at 17.7 ± 0.2°. The diffraction peak is present at 22.3 ± 0.2°; or at 21.0 ± 0.2°; or at 20.2 ± 0.2°; or at 23.1 ± 0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 diffraction peaks are included; more preferably, any 6, 7, or 8 diffraction peaks are included.

[0066] The X-ray powder diffraction pattern of crystal form C contains at least one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, and 9.2±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, and 17.7±0.2°, preferably two, three, four, or five.

[0067] The X-ray powder diffraction pattern of crystal form C optionally includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, 17.7±0.2°, 22.3±0.2°, and 21.0±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0068] More preferably, the X-ray powder diffraction pattern of crystal form C includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, 17.7±0.2°, 22.3±0.2°, 21.0±0.2°, 20.2±0.2°, 23.1±0.2°, 28.5±0.2°, 18.0±0.2°, and 19.9±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 locations.

[0069] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 3.

[0070] Table 3

[0071]

[0072] The crystalline form C of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 6 As shown; its DSC spectrum is basically as follows Figure 7 As shown.

[0073] In a further preferred embodiment of the present invention, the crystalline form D is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0074] The X-ray powder diffraction pattern of crystal form D shows a diffraction peak at 2θ = 23.3 ± 0.2°; or at 19.8 ± 0.2°; or at 16.9 ± 0.2°; or at 22.5 ± 0.2°; or at 14.7 ± 0.2°; or at 24.9 ± 0.2°; or at 20.6 ± 0.2°; or at 17.9 ± 0.2°. The diffraction peak is present at 0.2°; or at 15.0±0.2°; or at 8.1±0.2°; or at 15.6±0.2°; or at 7.6±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0075] The X-ray powder diffraction pattern of crystal form D contains at least one or more diffraction peaks located at 2θ of 23.3±0.2°, 19.8±0.2°, and 16.9±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 17.9±0.2°, and 15.0±0.2°, preferably two, three, four, or five.

[0076] The X-ray powder diffraction pattern of crystal form D may optionally include one or more diffraction peaks located at 2θ of 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.0±0.2°, and 8.1±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0077] More preferably, the X-ray powder diffraction pattern of crystal form D includes one or more diffraction peaks located at 2θ of 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.0±0.2°, 8.1±0.2°, 15.6±0.2°, 7.6±0.2°, 31.8±0.2°, 19.4±0.2°, and 21.2±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 locations.

[0078] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 4.

[0079] Table 4

[0080]

[0081]

[0082] The crystal form D of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 6 As shown; its DSC spectrum is basically as follows Figure 7 As shown.

[0083] In a further preferred embodiment of the present invention, the crystalline form E is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0084] The X-ray powder diffraction pattern of crystal form E shows a diffraction peak at 2θ of 19.0 ± 0.2°; or at 26.5 ± 0.2°; or at 4.8 ± 0.2°; or at 24.6 ± 0.2°; or at 9.1 ± 0.2°; or at 22.2 ± 0.2°; or at 25.2 ± 0.2°; or at 21.5 ± 0.2°. The diffraction peak is present at 2°; or at 5.2±0.2°; or at 14.1±0.2°; or at 20.9±0.2°; or at 15.2±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 diffraction peaks are included; more preferably, any 6, 7, or 8 diffraction peaks are included.

[0085] The X-ray powder diffraction pattern of crystal form E contains at least one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, and 4.8±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, and 21.5±0.2°, preferably two, three, four, or five.

[0086] The X-ray powder diffraction pattern of crystal form E optionally includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, 21.5±0.2°, 5.2±0.2°, and 14.1±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0087] The X-ray powder diffraction pattern of crystal form E includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, 21.5±0.2°, 5.2±0.2°, 14.1±0.2°, 20.9±0.2°, 15.2±0.2°, 18.6±0.2°, 17.6±0.2°, and 28.5±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0088] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 5.

[0089] Table 5

[0090]

[0091] The crystal form E of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 10 As shown; its DSC spectrum is basically as follows Figure 11 As shown.

[0092] In a further preferred embodiment of the invention, the crystalline form F is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0093] The X-ray powder diffraction pattern of crystal form F shows a diffraction peak at 2θ of 5.0 ± 0.2°; or at 16.2 ± 0.2°; or at 4.8 ± 0.2°; or at 19.8 ± 0.2°; or at 14.8 ± 0.2°; or at 18.3 ± 0.2°; or at 22.1 ± 0.2°; or at 22.5 ± 0.2°. The diffraction peak is present at 2°; or at 18.8±0.2°; or at 15.5±0.2°; or at 23.5±0.2°; or at 9.1±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0094] The X-ray powder diffraction pattern of crystal form F contains at least one or more diffraction peaks located at 2θ of 5.0±0.2°, 16.2±0.2°, and 4.8±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, and 22.5±0.2°, preferably two, three, four, or five.

[0095] The X-ray powder diffraction pattern of crystal form F may optionally include one or more diffraction peaks located at 2θ of 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, 22.5±0.2°, 18.8±0.2°, and 15.5±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0096] The X-ray powder diffraction pattern of crystal form F includes one or more diffraction peaks located at 2θ of 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, 22.5±0.2°, 18.8±0.2°, 15.5±0.2°, 23.5±0.2°, 9.1±0.2°, 25.0±0.2°, 19.0±0.2°, and 26.5±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0097] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 6.

[0098] Table 6

[0099]

[0100]

[0101] The crystalline form F of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 12 As shown; its DSC spectrum is basically as follows Figure 13 As shown.

[0102] In a further preferred embodiment of the invention, the crystalline form G is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0103] The X-ray powder diffraction pattern of crystal form G shows a diffraction peak at 2θ of 19.0 ± 0.2°; or at 26.5 ± 0.2°; or at 9.1 ± 0.2°; or at 25.0 ± 0.2°; or at 4.7 ± 0.2°; or at 14.0 ± 0.2°; or at 17.5 ± 0.2°; or at 23.5 ± 0.2°. The diffraction peak is present at 20.7 ± 0.2°; or at 21.3 ± 0.2°; or at 28.0 ± 0.2°; or at 22.2 ± 0.2°; or at 20.7 ± 0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 diffraction peaks are included; more preferably, any 6, 7, or 8 diffraction peaks are included.

[0104] The X-ray powder diffraction pattern of crystal form G contains at least one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, and 9.1±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, and 23.5±0.2°, preferably two, three, four, or five.

[0105] The X-ray powder diffraction pattern of crystal form G may optionally include one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, 23.5±0.2°, 21.3±0.2°, and 28.0±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0106] The X-ray powder diffraction pattern of crystal form G includes one or more diffraction peaks located at 2θ of 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, 23.5±0.2°, 21.3±0.2°, 28.0±0.2°, 22.2±0.2°, 20.7±0.2°, 24.2±0.2°, 23.1±0.2°, and 8.8±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0107] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 7.

[0108] Table 7

[0109]

[0110] The crystalline form G of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 14 As shown; its DSC spectrum is basically as follows Figure 15 As shown.

[0111] In a further preferred embodiment of the present invention, the crystalline form J is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0112] The X-ray powder diffraction pattern of crystal form J shows a diffraction peak at 2θ of 19.2 ± 0.2°; or at 14.1 ± 0.2°; or at 9.8 ± 0.2°; or at 22.4 ± 0.2°; or at 23.6 ± 0.2°; or at 15.9 ± 0.2°; or at 11.2 ± 0.2°; or at 16.8 ± 0.2°. The diffraction peak is present at 2°; or at 21.3±0.2°; or at 25.4±0.2°; or at 18.7±0.2°; or at 8.5±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0113] The X-ray powder diffraction pattern of crystal form J contains at least one or more diffraction peaks located at 2θ of 19.2±0.2°, 14.1±0.2°, and 9.8±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, and 16.8±0.2°, preferably two, three, four, or five.

[0114] The X-ray powder diffraction pattern of crystal form J may optionally include one or more diffraction peaks located at 2θ of 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, 16.8±0.2°, 21.3±0.2°, and 25.4±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 of these peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0115] The X-ray powder diffraction pattern of crystal form J includes one or more diffraction peaks located at 2θ of 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, 16.8±0.2°, 21.3±0.2°, 25.4±0.2°, 18.7±0.2°, 8.5±0.2°, 28.4±0.2°, 19.6±0.2°, and 20.4±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0116] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 8.

[0117] Table 8

[0118]

[0119]

[0120] The crystal form J of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 16 As shown; its DSC spectrum is basically as follows Figure 17 As shown.

[0121] In a further preferred embodiment of the present invention, the crystalline form K is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0122] The X-ray powder diffraction pattern of crystal form K shows a diffraction peak at 2θ = 25.2 ± 0.2°; or at 16.9 ± 0.2°; or at 20.5 ± 0.2°; or at 18.5 ± 0.2°; or at 23.8 ± 0.2°; or at 20.8 ± 0.2°; or at 10.2 ± 0.2°; or at 10.0 ± 0.2°. The diffraction peak is present at 0.2°; or at 7.8±0.2°; or at 11.6±0.2°; or at 24.2±0.2°; or at 21.8±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0123] The X-ray powder diffraction pattern of crystal form K contains at least one or more diffraction peaks located at 2θ of 25.2±0.2°, 16.9±0.2°, and 20.5±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, and 10.0±0.2°, preferably two, three, four, or five.

[0124] The X-ray powder diffraction pattern of crystal form K may optionally include one or more diffraction peaks located at 2θ of 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, 10.0±0.2°, 7.8±0.2°, and 11.6±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0125] The X-ray powder diffraction pattern of crystal form K includes one or more diffraction peaks located at 2θ of 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, 10.0±0.2°, 7.8±0.2°, 11.6±0.2°, 24.2±0.2°, 21.8±0.2°, 19.7±0.2°, 19.1±0.2°, and 15.4±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0126] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 9.

[0127] Table 9

[0128]

[0129]

[0130] The crystalline form K of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 18 As shown; its DSC spectrum is basically as follows Figure 19 As shown.

[0131] In a further preferred embodiment of the present invention, the crystalline form L is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0132] The X-ray powder diffraction pattern of crystal form L shows a diffraction peak at 2θ of 15.9 ± 0.2°; or at 19.6 ± 0.2°; or at 23.2 ± 0.2°; or at 18.1 ± 0.2°; or at 22.3 ± 0.2°; or at 4.8 ± 0.2°; or at 21.7 ± 0.2°; or at 15.3 ± 0.2°. The diffraction peak is present at 2°; or at 24.6±0.2°; or at 18.5±0.2°; or at 20.0±0.2°; or at 18.8±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0133] The X-ray powder diffraction pattern of crystal form L contains at least one or more diffraction peaks located at 2θ of 15.9±0.2°, 19.6±0.2°, and 23.2±0.2°, preferably two, more preferably three; optionally, it may further contain at least one peak located at 2θ of 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, and 15.3±0.2°, preferably two, three, four, or five.

[0134] The X-ray powder diffraction pattern of crystal form L optionally includes one or more diffraction peaks located at 2θ of 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, 15.3±0.2°, 24.6±0.2°, and 18.5±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 of these peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0135] The X-ray powder diffraction pattern of crystal form L includes one or more diffraction peaks located at 2θ of 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, 15.3±0.2°, 24.6±0.2°, 18.5±0.2°, 20.0±0.2°, 18.8±0.2°, 26.3±0.2°, 9.0±0.2°, and 17.2±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0136] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 10.

[0137] Table 10

[0138]

[0139] The present invention relates to the crystalline form L of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation, the X-ray powder diffraction pattern of which is basically as follows. Figure 20 As shown.

[0140] In a further preferred embodiment of the present invention, the crystalline form M is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0141] The X-ray powder diffraction pattern of crystal form M shows a diffraction peak at 2θ of 18.8 ± 0.2°; or at 26.3 ± 0.2°; or at 8.9 ± 0.2°; or at 4.5 ± 0.2°; or at 13.7 ± 0.2°; or at 24.7 ± 0.2°; or at 17.2 ± 0.2°; or at 21.9 ± 0.2°. The diffraction peak is present at 11.9 ± 0.2°; or at 27.7 ± 0.2°; or at 24.1 ± 0.2°; or at 21.0 ± 0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0142] The X-ray powder diffraction pattern of crystal form M contains at least one or more diffraction peaks located at 2θ of 18.8±0.2°, 26.3±0.2°, and 8.9±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, and 21.9±0.2°, preferably two, three, four, or five.

[0143] The X-ray powder diffraction pattern of crystal form M may optionally include one or more diffraction peaks located at 2θ of 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, 21.9±0.2°, 11.9±0.2°, and 27.7±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0144] The X-ray powder diffraction pattern of crystal form M includes one or more diffraction peaks located at 2θ of 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, 21.9±0.2°, 11.9±0.2°, 27.7±0.2°, 24.1±0.2°, 21.0±0.2°, 22.9±0.2°, 13.2±0.2°, and 8.5±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0145] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 11.

[0146] Table 11

[0147]

[0148]

[0149] The crystalline form M of the oxidized (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine of this invention, its X-ray powder diffraction pattern is basically as follows Figure 21 As shown.

[0150] In a further preferred embodiment of the present invention, the positions of the top ten diffraction peaks with the highest relative peak intensities in the X-ray powder diffraction patterns of crystal forms A, B, C, D, E, F, G, J, K, L, and M are respectively... Figure 1 , Figure 4 , Figure 6 , Figure 8 , Figure 10 , Figure 12 , Figure 14 , Figure 16 , Figure 18 , Figure 20 and Figure 21 The 2θ error of the diffraction peak at the corresponding position is ±0.2° to ±0.5°, with ±0.2° to ±0.3° being preferred and ±0.2° being the best option.

[0151] In a further preferred embodiment of the present invention, the method for determining the crystal form of the compound includes the following steps:

[0152] 1) Weigh an appropriate amount of free base and suspend it in a poor solvent. The preferred suspension density is 50-200 mg / mL.

[0153] 2) The suspension obtained above is shaken, preferably at a temperature of 0-60℃ and for a time of 0-10 days;

[0154] 3) The above suspension is rapidly centrifuged to remove the supernatant, and the remaining solid is dried to obtain the target product;

[0155] in:

[0156] The undesirable solvent is selected from acetone, ethyl acetate, isopropyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone or 3-pentanone, methyl tert-butyl ether, and water; preferably acetone, ethyl acetate, isopropyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, toluene, isopropanol, 2-butanone, 3-pentanone, methyl tert-butyl ether, and water.

[0157] In a further preferred embodiment of the present invention, the method for determining the crystal form of the compound includes the following steps:

[0158] 1) Weigh out an appropriate amount of free base and dissolve it in a good solvent;

[0159] 2) Optionally, add an antisolvent or a poor solvent to the solution obtained above, and stir until a solid precipitates;

[0160] 3) Optionally, add the antisolvent to the solution obtained above, centrifuge rapidly, remove the supernatant, and dry the remaining solid to obtain the target product;

[0161] in:

[0162] The benign solvent is selected from methanol, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone, or 3-pentanone; preferably methanol, dichloromethane, or tetrahydrofuran.

[0163] The unsuitable solvents are selected from heptane, water, methyl tert-butyl ether, toluene, and isopropyl ether.

[0164] In a further preferred embodiment of the present invention, the method for determining the crystal form of the compound includes the following steps: 1) Weighing an appropriate amount of free base and dissolving it by heating with a good solvent;

[0165] 2) Quickly place the solution obtained above at a low temperature and stir until a solid precipitates. The preferred temperature is -10 to 5°C.

[0166] 3) The above suspension was rapidly centrifuged to remove the supernatant. The remaining solid was placed in a vacuum drying oven at 40°C and dried to constant weight to obtain the target product.

[0167] in:

[0168] The benign solvent is selected from methanol, acetone, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone, or 3-pentanone; preferably 88% acetone. A further object of the present invention is to provide a pharmaceutical composition comprising a therapeutically effective amount of the crystal form of the compound of formula (I), and one or more pharmaceutically acceptable carriers or excipients.

[0169] The present invention also aims to provide the crystalline form of the compound of general formula (I) (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidized form, and the use of the pharmaceutical composition in the preparation of kinase inhibitor drugs.

[0170] The kinase inhibitor is a receptor tyrosine kinase inhibitor, preferably a HER2 inhibitor, an EGFR inhibitor, or an EGFR monoclonal antibody and its combination thereof, and more preferably a HER2 exon 20 mutant inhibitor, an EGFR exon 20 mutant inhibitor, or an EGFR exon 20 mutant monoclonal antibody and its combination thereof.

[0171] The purpose of this invention is to provide the crystal form of the compound represented by the general formula (I) and the use of the pharmaceutical composition in the treatment of cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases, preferably, the cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases are diseases mediated by HER2 exon 20 mutation and / or EGFR exon 20 mutation.

[0172] The present invention also aims to provide the crystalline form of the compound of general formula (I) (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidized form and the use of the pharmaceutical composition thereon in the treatment of cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases, preferably, the cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases are diseases mediated by HER2 exon 20 mutation and / or EGFR exon 20 mutation.

[0173] The cancers mentioned are selected from breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, liver cancer, solid tumors, glioma, glioblastoma, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

[0174] Third-generation EGFR inhibitors are primarily effective against EGFR-activating mutants and T790M resistance mutants. The compounds of this invention exhibit the following significant advantages over third-generation EGFR inhibitors in targeting EGFR and / or HER2 exon 20 insertion mutations:

[0175] 1. Significantly enhances the inhibitory activity of Ba / F3 EGFR mutant cell lines, with the preferred compounds showing 10-fold or even 20-fold higher activity;

[0176] 2. Improve the selectivity of the compound in inhibiting the proliferation of Ba / F3 EGFR mutant cell lines and A431 cell lines, with the preferred compound showing a selectivity of 3 times or more, or even 10 times.

[0177] 3. It also showed a significant advantage in in vivo drug efficacy and tumor inhibition rate in the mouse original B cell Ba / F3 EGFR-D770-N771ins_SVD xenograft model.

[0178] All contents contained in patents PCT / CN2020 / 097362 and PCT / CN2020 / 097369 are incorporated herein by reference.

[0179] The object of this invention is to provide an acid salt of the compound represented by general formula (I).

[0180]

[0181] in:

[0182] R1 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkylthio, haloalkoxy, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0183] R2 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, alkylthio, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, or heteroaryl.

[0184] Alternatively, any two R2 atoms can be linked with the carbon atoms they are attached to to form a cycloalkyl or heterocyclic group;

[0185] Ring A is selected from cycloalkyl, heterocyclic, aryl, or heteroaryl groups;

[0186] R a Selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, hydroxyalkyl, cyanoalkyl, alkoxy, haloalkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, -(CH2). n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0187] R aa and R bb Each is independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, alkyl, deuterated alkyl, haloalkyl, alkoxy, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic, aryl, or heteroaryl;

[0188] x is an integer between 0 and 4;

[0189] y is an integer between 0 and 4;

[0190] t is an integer between 0 and 1;

[0191] m is an integer between 0 and 2; and

[0192] n is an integer between 0 and 2.

[0193] The acid in the acid salt is selected from inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, or phosphoric acid; and the organic acid is selected from 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic acid, camphorsulfonic acid, aspartic acid, camphoric acid, gluconic acid, glucuronic acid, glutamic acid, isoascorbic acid, lactic acid, malic acid, mandelic acid, and pyroglutamic acid. Tartaric acid, dodecyl sulfuric acid, dibenzoyl tartaric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethyl sulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid;

[0194] y is 1, 2 or 3.

[0195] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein ring A is selected from 3-12 membered heterocyclic groups; preferably 3-8 membered heterocyclic groups; more preferably 3-8 membered monocyclic heterocyclic groups or fused heterocyclic groups containing 1-2 N or O atoms; and further preferably the following groups:

[0196]

[0197] In a preferred embodiment of the present invention, the crystal form represented by general formula (I), wherein R a Selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Hydroxyalkyl, C 1-6 Cyanoalkyl, C 1-6 Alkoxy, C 1-6 Haloalkoxy group, -(CH2) n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0198] Preferred elements include hydrogen, deuterium, halogens, hydroxyl groups, and C. 1-3 Alkyl, C 1-3 Haloalkyl, C 1-3 Hydroxyalkyl, C 1-3 Cyanoalkyl, C 1-3 Alkyl group, -(CH2) n OR aa -(CH2) n NR aa R bb -(CH2) n C(O)R aa Or -(CH2) n S(O) m R aa ;

[0199] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, hydroxyl, methyl, ethyl, isopropyl, -(CH2)2F, -CH2OH, -C(CH3)2OH, -CH2CN, -OCH2CH3, -CH2OCH3, -C(O)CH3, -S(O)2CH3, -N(CH3)2, -NCH3(CH2CH3) or

[0200] R aa and R bb Each is independently selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, haloalkoxy, C 2-6 alkenyl, C 2-6 alkynyl, cycloalkyl, heterocyclic, aryl or heteroaryl;

[0201] Preferred methyl, ethyl, or oxetine butyl groups;

[0202] x is an integer between 0 and 2.

[0203] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein R1 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy or C 1-6 Alkylthio;

[0204] Preferred elements: hydrogen, deuterium, halogens, C 1-6 Alkyl, C 1-3 Halogenated alkyl or C 1-3 Alkylthio;

[0205] More preferably, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, or methylthio;

[0206] The preferred compounds are chlorine, bromine, trifluoromethyl, or methylthio.

[0207] In a preferred embodiment of the present invention, the crystal form represented by general formula (I) wherein R2 is selected from hydrogen, deuterium, halogen, amino, nitro, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Deuterated alkyl, C 1-6 Haloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 2-6 alkenyl or C 2-6 alkynyl group;

[0208] Preferred elements: hydrogen, deuterium, halogens, C 1-6 Alkyl, C 1-3 Haloalkyl, C 1-3 Alkoxy or C 2-4 alkynyl group;

[0209] More preferably, hydrogen, deuterium, fluorine, chlorine, bromine, methyl, ethyl, isopropyl, trifluoromethyl, methoxy, or ethynyl;

[0210] The preferred radicals are hydrogen, fluorine, methyl, ethyl, trifluoromethyl, methoxy, or ethynyl.

[0211] Alternatively, any two R2 atoms can be linked with the carbon atoms they are attached to form a 3-8 membered heterocyclic group; preferably a 5-6 membered heterocyclic group containing 1-2 N or O atoms; more preferably a tetrahydrofuran group;

[0212] y is an integer between 0 and 3.

[0213] In a more preferred embodiment of the present invention, a crystalline form of the compound (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation is provided.

[0214] The compound is (6-((5-bromo-2-((2-methoxy-5-methyl-4-(4-((3aR,6aS)-tetrahydro-1H-furano[3,4-c]pyrrolo-5(3H)-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation;

[0215] (6-((5-chloro-2-((4-(4-(3-(dimethylamino)acetidin-1-yl)piperidin-1-yl)-5-ethyl-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation;

[0216] 2-(1-(1-(4-((5-bromo-4-((5-(dimethylphospho)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-yl)acetidin-3-yl)acetonitrile; or,

[0217] Acidic salt of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation.

[0218] The acid salt of the compound represented by general formula (I) is characterized in that the acid salt oxidized by (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine is in the crystalline form, preferably in the phosphate, hydrochloride or methanesulfonate form.

[0219] The X-ray powder diffraction pattern of phosphate crystal form A shows diffraction peaks at 2θ of 13.8 ± 0.2°; or at 23.4 ± 0.2°; or at 21.0 ± 0.2°; or at 25.4 ± 0.2°; or at 20.3 ± 0.2°; or at 10.1 ± 0.2°; or at 9.5 ± 0.2°; or at 21.4 ± 0.2°. The diffraction peak is present at 0.2°; or at 17.0±0.2°; or at 24.4±0.2°; or at 22.9±0.2°; or at 20.5±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0220] As a specific example, the X-ray powder diffraction pattern of phosphate crystal form A contains at least one or more diffraction peaks located at 2θ of 13.8±0.2°, 23.4±0.2°, and 21.0±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, and 21.4±0.2°, preferably two, three, four, or five.

[0221] As a specific example, the X-ray powder diffraction pattern of phosphate crystal form A optionally includes one or more diffraction peaks located at 2θ of 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, 21.4±0.2°, 17.0±0.2°, and 24.4±0.2°; preferably, it includes at least any 2-3 peaks, or 4-5 peaks, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0222] As a specific example, the X-ray powder diffraction pattern of phosphate crystal form A includes one or more diffraction peaks located at 2θ of 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, 21.4±0.2°, 17.0±0.2°, 24.4±0.2°, 22.9±0.2°, 20.5±0.2°, 22.6±0.2°, 17.5±0.2°, and 13.5±0.2°, preferably including any 4, 5, 6, 8, or 10 diffraction peaks therein;

[0223] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 12.

[0224] Table 12

[0225]

[0226]

[0227] The phosphate crystal form A of the (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation of this invention has an X-ray powder diffraction pattern that is basically as follows: Figure 22As shown; its DSC spectrum is basically as follows Figure 23 As shown.

[0228] In a further preferred embodiment of the present invention, the hydrochloride crystal form A is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0229] The X-ray powder diffraction pattern of hydrochloride crystal form A shows a diffraction peak at 2θ = 22.7 ± 0.2°; or at 19.4 ± 0.2°; or at 25.3 ± 0.2°; or at 22.3 ± 0.2°; or at 25.7 ± 0.2°; or at 8.2 ± 0.2°; or at 17.9 ± 0.2°; or at 29.1 ± 0.2°. The diffraction peak is present at 0.2°; or at 14.3±0.2°; or at 13.6±0.2°; or at 16.3±0.2°; or at 30.8±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0230] The X-ray powder diffraction pattern of hydrochloride crystal form A contains at least one or more diffraction peaks located at 2θ of 22.7±0.2°, 19.4±0.2°, and 25.3±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, and 29.1±0.2°, preferably two, three, four, or five.

[0231] The X-ray powder diffraction pattern of hydrochloride crystal form A optionally includes one or more diffraction peaks located at 2θ of 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, 29.1±0.2°, 14.3±0.2°, and 13.6±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 of these peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0232] The X-ray powder diffraction pattern of hydrochloride crystal form A includes one or more diffraction peaks located at 2θ of 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, 29.1±0.2°, 14.3±0.2°, 13.6±0.2°, 16.3±0.2°, 30.8±0.2°, 23.8±0.2°, 20.7±0.2°, and 27.5±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0233] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 13.

[0234] Table 13

[0235]

[0236] The present invention relates to the hydrochloride crystal form A of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation, the X-ray powder diffraction pattern of which is basically as follows. Figure 24 As shown; its DSC spectrum is basically as follows Figure 25 As shown.

[0237] In a further preferred embodiment of the invention, the methanesulfonate crystal form A is obtained by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine.

[0238] The X-ray powder diffraction pattern of methanesulfonate crystal form A shows a diffraction peak at 2θ = 20.9 ± 0.2°; or at 6.2 ± 0.2°; or at 13.4 ± 0.2°; or at 22.4 ± 0.2°; or at 25.6 ± 0.2°; or at 14.8 ± 0.2°; or at 18.9 ± 0.2°; or at 10.3 ± The diffraction peak is present at 0.2°; or at 18.3±0.2°; or at 24.6±0.2°; or at 8.2±0.2°; or at 12.8±0.2°; preferably, any 2-5, 3-5, 3-6, 3-8, 5-8, or 6-8 of the above diffraction peaks are included; more preferably, any 6, 7, or 8 of the above diffraction peaks are included.

[0239] The X-ray powder diffraction pattern of methanesulfonate crystal form A contains at least one or more diffraction peaks located at 2θ of 20.9±0.2°, 6.2±0.2°, and 13.4±0.2°, preferably two, more preferably three; optionally, it may further contain at least one diffraction peak located at 2θ of 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, and 10.3±0.2°, preferably two, three, four, or five.

[0240] The X-ray powder diffraction pattern of methanesulfonate crystal form A optionally includes one or more diffraction peaks located at 2θ of 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, 10.3±0.2°, 18.3±0.2°, and 24.6±0.2°; preferably, it includes at least 2-3, 4-5, or 6-7 peaks; more preferably, it includes any 2, 3, 4, 5, 6, or 7 peaks.

[0241] More preferably, the X-ray powder diffraction pattern of methanesulfonate crystal form A includes one or more diffraction peaks located at 2θ of 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, 10.3±0.2°, 18.3±0.2°, 24.6±0.2°, 8.2±0.2°, 12.8±0.2°, 31.9±0.2°, 9.5±0.2°, and 29.6±0.2°. Preferably, it includes diffraction peaks at any of the selected 4, 5, 6, 8, or 10 of these peaks.

[0242] The characteristic X-ray diffraction peaks, expressed in terms of 2θ angle and interplanar spacing d, using Cu-Kα radiation are shown in Table 14.

[0243] Table 14

[0244]

[0245]

[0246] The methanesulfonate crystal form A of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation of this invention has an X-ray powder diffraction pattern that is basically as follows: Figure 26 As shown; its DSC spectrum is basically as follows Figure 27 As shown.

[0247] In a further preferred embodiment of the present invention, the positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction patterns of phosphate crystal form A, hydrochloride crystal form A, and methanesulfonate crystal form A are respectively... Figure 1 , Figure 3 and Figure 5 The 2θ error of the diffraction peak at the corresponding position is ±0.2° to ±0.5°, with ±0.2° to ±0.3° being preferred and ±0.2° being the best option.

[0248] 1) Weigh out an appropriate amount of free base and dissolve it in a good solvent;

[0249] 2) Weigh an appropriate amount of the counterion acid and dissolve it in an organic solvent; the amount of counterion acid is preferably 1.2 equivalents;

[0250] 3) Combine the two solutions mentioned above and stir to precipitate, or add a poor solvent and stir to precipitate;

[0251] 4) Rapid centrifugation or static drying to obtain the target product;

[0252] in:

[0253] The beneficial solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, acetonitrile, 2-butanone, 3-pentanone, tetrahydrofuran, N,N-dimethylformamide, N,N-dimethylacetamide, or N-methylpyrrolidone; preferably methanol or ethanol.

[0254] The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol, or N,N-dimethylformamide; preferably methanol, ethanol, or acetonitrile; the above-mentioned benign solvents and organic solutions must be miscible when used.

[0255] The unsuitable solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone, or acetonitrile; preferably water, methyl tert-butyl ether, or isopropyl ether.

[0256] The aforementioned counterionic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic 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, dodecyl sulfate, and dibenzoyl tartaric acid. Ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethylsulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-methylbenzenesulfonic acid, or L-malic acid; preferably phosphoric acid, hydrochloric acid, or methanesulfonic acid; most preferably phosphoric acid.

[0257] In a further preferred embodiment of the present invention, the method for preparing the acid salt of the compound includes the following steps:

[0258] 1) Weigh an appropriate amount of free base and suspend it in a poor solvent;

[0259] 2) Weigh an appropriate amount of the counterion acid and dissolve it in an organic solvent; the amount of counterion acid is preferably 1.2 equivalents;

[0260] 3) Combine the two solutions and stir to dissolve them. Continue stirring to precipitate, or add a poor solvent and stir to precipitate.

[0261] 4) Rapid centrifugation or static drying to obtain the target product;

[0262] in:

[0263] The undesirable solvent is selected from ethyl acetate, acetone, dichloromethane, acetonitrile, tetrahydrofuran, 2-methyltetrahydrofuran, 2-butanone, 3-pentanone, and 1,4-dioxane; preferably ethyl acetate, acetone, acetonitrile, and 2-methyltetrahydrofuran.

[0264] The organic solvent is selected from methanol, ethanol, ethyl acetate, dichloromethane, acetone, n-hexane, petroleum ether, benzene, toluene, chloroform, acetonitrile, carbon tetrachloride, dichloroethane, tetrahydrofuran, 2-butanone, 3-pentanone, heptane, methyl tert-butyl ether, isopropyl ether, 1,4-dioxane, tert-butanol, or N,N-dimethylformamide; preferably methanol, ethanol, or acetonitrile; the above-mentioned benign solvents and organic solutions must be miscible when used.

[0265] The unsuitable solvent is selected from heptane, water, methyl tert-butyl ether, toluene, isopropyl ether, ethyl acetate, acetone, or acetonitrile; preferably water, methyl tert-butyl ether, or isopropyl ether.

[0266] The aforementioned counterionic acid is selected from hydrochloric acid, sulfuric acid, nitric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, phosphoric acid, 2,5-dihydroxybenzoic acid, 1-hydroxy-2-naphtholic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, acetoxyxamic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, benzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, decanoic acid, hexanoic acid, caprylic acid, cinnamic acid, citric acid, cyclohexanesulfonic 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, dodecyl sulfate, and dibenzoyl tartaric acid. Ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactobionic acid, gentian acid, glutaric acid, 2-ketoglutaric acid, glycolic acid, hippuric acid, hydroxyethylsulfonic acid, lactobionic acid, ascorbic acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, methanesulfonic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, dihydroxynaphthalic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, thiocyanate, undecanoic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluenesulfonic acid, or L-malic acid; preferably phosphoric acid, hydrochloric acid, or methanesulfonic acid; most preferably phosphoric acid. The present invention also aims to provide a pharmaceutical composition comprising a therapeutically effective amount of an acid salt of the compound of the general formula (I), and one or more pharmaceutically acceptable carriers or excipients.

[0267] The present invention also aims to provide an acid salt of the compound of general formula (I) (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation, and the use of the pharmaceutical composition in the preparation of kinase inhibitor drugs.

[0268] The kinase inhibitor is a receptor tyrosine kinase inhibitor, preferably a HER2 inhibitor, an EGFR inhibitor, or an EGFR monoclonal antibody and its combination thereof, and more preferably a HER2 exon 20 mutant inhibitor, an EGFR exon 20 mutant inhibitor, or an EGFR exon 20 mutant monoclonal antibody and its combination thereof.

[0269] The purpose of this invention is to provide an acid salt of the compound represented by general formula (I) and the use of the pharmaceutical composition thereon in the treatment of cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases, preferably, the cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases are diseases mediated by HER2 exon 20 mutations and / or EGFR exon 20 mutations.

[0270] The present invention also aims to provide an acid salt of the compound of general formula (I) (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[B][1,4]dioxin-5-yl)dimethylphosphine oxidation and the use of the pharmaceutical composition thereof in the treatment of cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases, preferably, the cancer, inflammation, chronic liver disease, diabetes, cardiovascular disease and AIDS-related diseases are diseases mediated by HER2 exon 20 mutation and / or EGFR exon 20 mutation.

[0271] The cancers mentioned are selected from breast cancer, cervical cancer, colon cancer, lung cancer, stomach cancer, rectal cancer, pancreatic cancer, brain cancer, liver cancer, solid tumors, glioma, glioblastoma, leukemia, lymphoma, myeloma, and non-small cell lung cancer.

[0272] Third-generation EGFR inhibitors are primarily effective against EGFR-activating mutants and T790M resistance mutants. The compounds of this invention exhibit the following significant advantages over third-generation EGFR inhibitors in targeting EGFR and / or HER2 exon 20 insertion mutations:

[0273] 4. Significantly enhances the inhibitory activity of Ba / F3 EGFR mutant cell lines, with the preferred compounds showing 10-fold or even 20-fold higher activity;

[0274] 5. Improve the selectivity of the compound in inhibiting the proliferation of Ba / F3 EGFR mutant cell lines and A431 cell lines, with the preferred compound showing a selectivity of 3 times or more, or even 10 times.

[0275] 3. It also showed a significant advantage in in vivo drug efficacy and tumor inhibition rate in the mouse original B cell Ba / F3 EGFR-D770-N771ins_SVD xenograft model.

[0276] Detailed description of the invention

[0277] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0278] "Hydroxy" refers to the -OH group. "Halogen" refers to fluorine, chlorine, bromine, or iodine. "Amino" refers to -NH₂. "Cyano" refers to -CN. "Nitro" refers to -NO₂. "Carboxyl" refers to -C(O)OH. "THF" refers to tetrahydrofuran. "EtOAc" refers to ethyl acetate. "MeOH" refers to methanol. "DMF" refers to N,N-dimethylformamide. "DIPEA" refers to diisopropylethylamine. "TFA" refers to trifluoroacetic acid. "MeCN" refers to acetonitrile. "DMA" refers to N,N-dimethylacetamide. "Et₂O" refers to diethyl ether. "DCE" refers to 1,2-dichloroethane. "DIPEA" refers to N,N-diisopropylethylamine. "NBS" refers to N-bromosuccinimide. "NIS" refers to N-iodosuccinimide. "Cbz-Cl" refers to benzyl chloroformate. “Pd2(dba)3” refers to tris(dibenzylacetone)dipalladium. “Dppf” refers to 1,1'-bis(diphenylphosphine)ferrocene. “HATU” refers to 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate. “KHMDS” refers to potassium hexamethyldisilamide. “LiHMDS” refers to lithium bis(trimethylsilylamine). “MeLi” refers to methyllithium. “n-BuLi” refers to n-butyllithium. “NaBH(OAc)3” refers to sodium triacetoxyborohydride. “DMAP” refers to 4-dimethylaminopyridine. “SEM-Cl” refers to chloromethyltrimethylsilylethyl ether. “Xantphos” refers to 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene. “DCM” refers to dichloromethane.

[0279] The different terms such as "X is selected from A, B, or C", "X is selected from A, B, and C", "X is A, B, or C", and "X is A, B, and C" all express the same meaning, that is, X can be any one or more of A, B, and C.

[0280] All hydrogen atoms described in this invention can be replaced by their isotope deuterium, and any hydrogen atom in the compounds of the embodiments of this invention can also be replaced by a deuterium atom.

[0281] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and the exertion of its biological activity.

[0282] "Medicinal salts" refer to the salts of the compounds of this invention, which are safe and effective when used in mammals and have the appropriate biological activity. Attached Figure Description

[0283] Figure 1 The image shows the XRPD diagram of free alkali crystal form A.

[0284] Figure 2 The DSC diagram shows the free alkali crystal form A.

[0285] Figure 3 TGA illustration of free alkali crystal form A.

[0286] Figure 4 The image shows the XRPD diagram of free base crystal form B.

[0287] Figure 5 The DSC diagram shows the free alkali crystal form B.

[0288] Figure 6 The image shows the XRPD diagram of free base crystal form C.

[0289] Figure 7 The image shows the DSC diagram of the free basal crystal form C.

[0290] Figure 8 The image shows the XRPD diagram of the free base crystal form D.

[0291] Figure 9 The DSC diagram shows the free alkali crystal form D.

[0292] Figure 10 The image shows the XRPD diagram of the free base crystal form E.

[0293] Figure 11 The image shows the DSC diagram of the free base crystal form E.

[0294] Figure 12 The image shows the XRPD diagram of the free basal crystal form F.

[0295] Figure 13 The image shows the DSC diagram of the free basal form F.

[0296] Figure 14 The image shows the XRPD diagram of the free basal crystal form G.

[0297] Figure 15 DSC diagram of free base crystal form G.

[0298] Figure 16 The image shows the XRPD diagram of the free alkali crystal form J.

[0299] Figure 17 The DSC diagram shows the free alkali crystal form J.

[0300] Figure 18 The image shows the XRPD diagram of the free alkali crystal form K.

[0301] Figure 19 The image shows the DSC diagram of the free base crystal form K.

[0302] Figure 20 The image shows the XRPD diagram of the free base crystal form L.

[0303] Figure 21 The image shows the XRPD diagram of the free alkali crystal form M.

[0304] Figure 22 This is an XRPD diagram of phosphate crystal form A.

[0305] Figure 23 This is a DSC diagram of phosphate crystal form A.

[0306] Figure 24 This is an XRPD diagram of hydrochloride crystal form A.

[0307] Figure 25 This is a DSC diagram of hydrochloride crystal form A.

[0308] Figure 26 This is an XRPD diagram of methanesulfonate crystal form A.

[0309] Figure 27 This is a DSC diagram of methanesulfonate crystal form A.

[0310] In the attached diagram, intensity represents intensity; Theta represents θ; and counts represents counts. Detailed Implementation

[0311] The present invention is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the present invention.

[0312] I. Preparation of Compounds

[0313] Example

[0314] The structures of the compounds of this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), and deuterated chloroform (CDCl3) as solvents, and tetramethylsilane (TMS) as the internal standard.

[0315] LC-MS was performed using an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18 150×4.6 mm column).

[0316] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The standard size for TLC is 0.15mm to 0.20mm, while the standard size for TLC separation and purification is 0.4mm to 0.5mm. Column chromatography generally uses 200-300 mesh Yantai Huanghai silica gel as the carrier.

[0317] The starting materials used in the embodiments of the present invention are known and commercially available, or can be synthesized using methods known in the art.

[0318] Unless otherwise specified, all reactions in this invention are carried out under continuous magnetic stirring, in a dry nitrogen or argon atmosphere, using a dry solvent, and the reaction temperature is expressed in degrees Celsius.

[0319] Preparation of intermediates 1

[0320] Preparation of (6-amino-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation

[0321]

[0322] Step 1: Preparation of 2-bromo-6-methoxy-3-nitrophenol

[0323]

[0324] NBS (2.1 g, 11.8 mmol) was added to a DCM (20 mL) solution of 2-methoxy-5-nitrophenol (2 g, 11.8 mmol). After stirring at room temperature for one hour, CH2Cl2 and water were added to the reaction mixture. The organic phase was concentrated under reduced pressure and separated by column chromatography to obtain the title compound 2-bromo-6-methoxy-3-nitrophenol (1.5 g, yield: 51%).

[0325] MS m / z (ESI): 245.9 [MH] - .

[0326] Step 2: Preparation of 3-bromo-4-nitrobenzene-1,2-diol

[0327]

[0328] At -78°C, a solution of 2-bromo-6-methoxy-3-nitrophenol (500 mg, 2.0 mmol) in dichloromethane (5 mL) was added, followed by the addition of a solution of BBr3 (1 M, 2.6 mL, 2.6 mmol) in dichloromethane. After stirring for 2 hours, the mixture was slowly brought to room temperature and stirred overnight. Upon cooling to 0°C, MeOH (5 mL) was slowly added dropwise to the reaction mixture. The organic phase was concentrated under reduced pressure and separated by column chromatography to obtain the title compound 3-bromo-4-nitrobenzene-1,2-diol (410 mg, yield: 87%).

[0329] MS m / z (ESI): 231.9 [MH] - .

[0330] Step 3: Preparation of 5-bromo-6-nitro-2,3-dihydrobenzo[b][1,4]dioxin

[0331]

[0332] 3-Bromo-4-nitrobenzene-1,2-diol (410 mg, 1.75 mmol), potassium carbonate (0.73 g, 5.26 mmol), and 1,2-dibromoethane (1.32 g, 7.0 mmol) were mixed in DMF (5 mL) and stirred overnight at 90 °C. After cooling, the mixture was diluted with a large amount of ethyl acetate. The organic phase was washed several times with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography yielded the title compound, 5-bromo-6-nitro-2,3-dihydrobenzo[b][1,4]dioxin (200 mg, yield: 44%).

[0333] MS m / z (ESI): 257.9 [MH] - .

[0334] Step 4: Preparation of 5-bromo-2,3-dihydrobenzo[b][1,4]dioxin-6-amine

[0335]

[0336] 5-Bromo-6-nitro-2,3-dihydrobenzo[b][1,4]dioxin (200 mg, 0.77 mmol) was dissolved in ethanol (9 mL) and water (3 mL), and reduced iron powder (343 mg, 6.1 mmol) and ammonium chloride (82 mg, 1.5 mmol) were added. The mixture was refluxed for 3 h. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give the title compound 5-bromo-2,3-dihydrobenzo[b][1,4]dioxin-6-amine (170 mg, yield: 96%).

[0337] MS m / z(ESI): 230.2 [M+H] + .

[0338] Step 5: Preparation by oxidation of (6-amino-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine

[0339]

[0340] 5-Bromo-2,3-dihydrobenzo[b][1,4]dioxin-6-amine (0.16 g, 0.7 mmol), dimethylphosphine oxide (108 mg, 1.39 mmol), and potassium phosphate (295 mg, 1.39 mmol) were mixed in N,N-dimethylformamide (5 mL), and palladium acetate (31 mg, 0.14 mmol) and Xantphos (161 mg, 0.28 mmol) were added. The mixture was deoxygenated with N2 for 5 minutes, and then microwaved to 145 °C for 3 hours. After cooling to room temperature, the organic solvent was concentrated under reduced pressure, and column chromatography was used to separate the contents to obtain the title compound (6-amino-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxide (83 mg, yield: 52%).

[0341] 1 H NMR (400MHz, CDCl3) δ1.72(s,3H),1.75(s,3H),4.09-4.13(m,2H),4.15-4.23(m,2H),5.41-5.85(m,2H),6.07-6.15(m,1H),6.72(d,J=6.8,1H);

[0342] MS m / z(ESI): 228.2 [M+H] + .

[0343] Preparation of intermediates 2

[0344] 2-Methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)aniline

[0345]

[0346] Step 1: Preparation of 8-(5-methoxy-2-methyl-4-nitrophenyl)-1,4-dioxa-8-azaspiro[4.5]decane

[0347]

[0348] K₂CO₃ (1.6 g, 11.9 mmol) was added to a DMSO (15 mL) solution of 1-fluoro-5-methoxy-2-methyl-4-nitrobenzene (1.1 g, 5.9 mmol) and 4-piperidinone ethylene glycol (3.4 g, 23.9 mmol), and the mixture was stirred overnight at 120 °C. The reaction solution was cooled to room temperature, and the organic solvent was concentrated under reduced pressure. The solution was then separated by column chromatography to obtain the title compound 8-(5-methoxy-2-methyl-4-nitrophenyl)-1,4-dioxa-8-azaspiro[4.5]decane (1.3 g, yield: 71%).

[0349] MS m / z (ESI): 309.2 [M+H] + .

[0350] Step 2: Preparation of 2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)aniline

[0351]

[0352] 8-(5-methoxy-2-methyl-4-nitrophenyl)-1,4-dioxa-8-azaspiro[4.5]decane (500 mg, 1.62 mmol) was dissolved in methanol (10 mL) and tetrahydrofuran (3 mL), and Pd / C (100 mg) was added. The mixture was stirred at room temperature for 5 h under a hydrogen atmosphere. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give the title compound 2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)aniline (433 mg, yield: 96%).

[0353] MS m / z(ESI): 279.2 [M+H] + .

[0354] Example 1

[0355] Preparation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation

[0356]

[0357] Step 1: Preparation by oxidation of (6-((5-bromo-2-chloropyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine

[0358]

[0359] At room temperature, 5-bromo-2,4-dichloropyrimidine (2.27 g, 10 mmol), (6-amino-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation (2.27 g, 10 mmol), and potassium phosphate (2.76 g, 20 mmol) were mixed in tert-amyl alcohol (20 mL) and reacted in a microwave oven at 90 °C for 1 h. After cooling to room temperature, the organic solvent was concentrated under reduced pressure and then separated by column chromatography to obtain the title compound (6-((5-bromo-2-chloropyrimidine-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation (3.25 g, yield: 78%).

[0360] 1 H NMR (400MHz, DMSO-d) 6 )δ1.81(s,3H),1.85(s,3H),4.24-4.39(m,4H),7.13(d,J=9.2Hz,1H),7.89-7.98(m,1H),8.44(d,J=1.8Hz,1H),12.26(s,1H);

[0361] MS m / z (ESI): 417.9 [M+H] + .

[0362] Step 2: Preparation by oxidation of (6-((5-bromo-2-((2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)phenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine

[0363]

[0364] At room temperature, (6-((5-bromo-2-chloropyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidase (1.0 g, 2.4 mmol), 2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)aniline (1.0 g, 3.6 mmol), and p-toluenesulfonic acid (0.62 g, 3.6 mmol) were mixed in ethylene glycol (40 mL) and the mixture was heated to 90 °C and reacted for 2 h. The reaction was cooled to room temperature, saturated sodium bicarbonate aqueous solution was added, and the mixture was extracted with ethyl acetate. The organic phase was separated and washed with saturated brine. The organic phase was dried with anhydrous sodium sulfate, filtered to remove the drying agent, and the organic solvent was concentrated under reduced pressure. Column chromatography was used to separate the title compound (6-((5-bromo-2-((2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)phenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxide (1.34 g, yield: 85%).

[0365] 1 H NMR (400MHz, DMSO-d) 6 )δ1.74-1.85(m,10H),2.14(s,3H),2.91(t,J=5.4Hz,4H),3.77(s,3H),3.93(s,4H),4.24(s,2H),4.32(s ,2H),6.73(s,1H),6.81(d,J=9.2Hz,1H),7.46(s,1H),7.94(d,J=15.0Hz,2H),8.10(s,1H),11.62(s,1H);

[0366] MS m / z (ESI): 660.2 [M+H] + .

[0367] Step 3: Preparation of 1-(4-((5-bromo-4-((5-(dimethylphospho)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-one

[0368]

[0369] At room temperature, (6-((5-bromo-2-((2-methoxy-5-methyl-4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)phenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxide (1.34 g, 2.03 mmol) was mixed in acetic acid / water (12 mL / 12 mL) and heated to 90 °C for 2 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and extracted with ethyl acetate after adding a saturated aqueous sodium bicarbonate solution. The organic phases were combined and dried over anhydrous sodium sulfate. After filtering with the desiccant, the organic solvent was concentrated under reduced pressure and separated by column chromatography to obtain the title compound 1-(4-((5-bromo-4-((5-(dimethylphospho)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-one (1.2 g, yield: 96%).

[0370] MS m / z (ESI): 616.2 [M+H] + .

[0371] Step 4: Preparation by oxidation of (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine

[0372]

[0373] At room temperature, 1-(4-((5-bromo-4-((5-(dimethylphospho)-2,3-dihydrobenzo[b][1,4]dioxin-6-yl)amino)pyrimidin-2-yl)amino)-5-methoxy-2-methylphenyl)piperidin-4-one (0.8 g, 1.3 mmol), 3-(methoxymethyl)acetidine trifluoroacetate (0.42 g, 1.95 mmol) and acetic acid (0.1 mL) were mixed in dichloroethane (20 mL), stirred for 30 minutes, and then sodium triacetoxyborohydride (0.55 g, 2.6 mmol) was added and stirred overnight at room temperature. Add saturated sodium bicarbonate aqueous solution, extract with dichloromethane, dry the organic phase with anhydrous sodium sulfate, filter the desiccant and concentrate the organic solvent under reduced pressure, and separate by column chromatography to obtain the title compound (6-((5-bromo-2-((2-methoxy-4-(4-(3-(methoxymethyl)acetidin-1-yl)piperidin-1-yl)-5-methylphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation (0.56 g, yield: 62%).

[0374] 1H NMR (400MHz, DMSO-d) 6 )δ1.26-1.38(m,2H),1.69-1.77(m,2H),1.78(s,3H),1.81(s,3H),2.07-2.22(m,4H),2. 56-2.66(m,3H),2.83-2.92(m,1H),2.96-3.05(m,2H),3.15-3.19(m,2H),3.23-3.27(m, 3H),3.42-3.46(m,2H),3.76(s,3H),4.07-4.13(m,1H),4.24(s,2H),4.32(s,2H),6.69( s,1H),6.80(d,J=9.0Hz,1H),7.43(s,1H),7.91-7.98(m,2H),8.10(s,1H),11.62(s,1H);

[0375] MS m / z (ESI): 701.2 [M+H] + .

[0376] Example 2

[0377] Preparation of (6-((5-bromo-2-((4-(4-(3-(dimethylamino)acetidin-1-yl)piperidin-1-yl)-5-ethyl-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine oxidation

[0378]

[0379] Step 1: Preparation of tert-butyl-4-(3-(dimethylamino)acetidin-1-yl)piperidine-1-carboxylic acid ester

[0380]

[0381] At room temperature, tert-butyl-4-carbonylpiperidin-1-carboxylic acid ester (500 mg, 2.51 mmol) and N,N-dimethylacetidine-3-amine (302 mg, 3.01 mmol) were dissolved in 1,2-dichloroethane (15 mL), 2 drops of acetic acid were added, and the mixture was stirred for 5 minutes. Sodium triacetoxyborohydride (1.06 g, 5.02 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction was then quenched with saturated sodium bicarbonate solution, and the mixture was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered through the drying agent, concentrated under reduced pressure, and separated by column chromatography to obtain the title compound tert-butyl-4-(3-(dimethylamino)acetidine-1-yl)piperidin-1-carboxylic acid ester (610 mg, yield: 86%).

[0382] 1 H NMR (400MHz, CDCl3) δ1.14-1.23(m,2H),1.44(s,9H),1.62-1.70(m,2H),2.12(s,6H),2.81-2.89(m,6H),3.48-3.53(m,2H),4.03-3.87(m,2H);

[0383] MS m / z(ESI): 284.1 [M+H] + .

[0384] Step 2: Preparation of N,N-dimethyl-1-(piperidin-4-yl)acetidine-3-amine

[0385]

[0386] At room temperature, tert-butyl-4-(3-(dimethylamino)acetidin-1-yl)piperidine-1-carboxylic acid ester (610 mg, 2.16 mmol) was dissolved in dioxane hydrochloride (10 mL), stirred overnight at room temperature, and the organic solvent was concentrated under reduced pressure to obtain crude N,N-dimethyl-1-(piperidine-4-yl)acetidin-3-amine, which was directly used in the next step of the reaction.

[0387] MS m / z (ESI): 184.1 [M+H] + .

[0388] Step 3: Preparation of 1-(1-(2-bromo-5-methoxy-4-nitrophenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine

[0389]

[0390] At room temperature, crude 1-bromo-2-fluoro-4-methoxy-5-nitrobenzene (300 mg, 1.2 mmol), crude N,N-dimethyl-1-(piperidin-4-yl)acetidine-3-amine (220 mg, 1.2 mmol), and potassium carbonate (497 mg, 3.6 mmol) were dissolved in N,N-dimethylformamide (8 mL). The mixture was heated to 60 °C and stirred overnight. Water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered off the drying agent, concentrated the organic solvent under reduced pressure, and separated by column chromatography to obtain the title compound 1-(1-(2-bromo-5-methoxy-4-nitrophenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine (430 mg, yield: 87%).

[0391] MS m / z(ESI): 413.1 [M+H] + .

[0392] Step 4: Preparation of 1-(1-(5-methoxy-4-nitro-2-vinylphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine

[0393]

[0394] At room temperature, 1-(1-(2-bromo-5-methoxy-4-nitrophenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine (430 mg, 1.04 mmol), potassium vinyltrifluoroborate (279 mg, 2.08 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (76 mg, 0.104 mmol), and cesium carbonate (1.01 g, 3.12 mmol) were dissolved in dioxane / water (10 mL / 1.5 mL), purged with nitrogen three times, heated to 90 °C and stirred overnight, water was added to the reaction system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried with anhydrous sodium sulfate, filtered to remove the drying agent, concentrated under reduced pressure, and separated by column chromatography to obtain the title compound 1-(1-(5-methoxy-4-nitro-2-vinylphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine (230 mg, yield: 61%).

[0395] MS m / z(ESI): 361.1 [M+H] + .

[0396] Step 5: Preparation of 1-(1-(4-amino-2-ethyl-5-methoxyphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine

[0397]

[0398] 1-(1-(5-methoxy-4-nitro-2-vinylphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine (230 mg, 0.64 mmol) was dissolved in methanol (10 mL) at room temperature, purged three times with nitrogen, and palladium / carbon (46 mg) was added. The mixture was stirred overnight at room temperature under a hydrogen atmosphere. The catalyst was removed by filtration, and the organic solvent was concentrated under reduced pressure to give the title compound 1-(1-(4-amino-2-ethyl-5-methoxyphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine (210 mg, yield: 98%).

[0399] MS m / z(ESI): 333.1 [M+H] + .

[0400] Referring to the second step of Example 1, the prepared 1-(1-(4-amino-2-ethyl-5-methoxyphenyl)piperidin-4-yl)-N,N-dimethylacetidine-3-amine was oxidized with (6-(((5-bromo-2-chloropyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine to prepare the target product (6-((5-bromo-2-((4-(4-(3-(dimethylamino)acetidine-1-yl)piperidin-1-yl)-5-ethyl-2-methoxyphenyl)amino)pyrimidin-4-yl)amino)-2,3-dihydrobenzo[b][1,4]dioxin-5-yl)dimethylphosphine.

[0401] 1 H NMR (400MHz, CD3OD) δ0.98-1.07(m,3H),1.47-1.60(m,2H),1.87(s,3H),1.91(s,3H),1.96-2. 05(m,2H),2.24(s,6H),2.50-2.59(m,2H),2.71-2.83(m,4H),3.03-3.10(m,2H),3.12-3.19(m, 1H),3.47-3.58(m,2H),3.84(d,J=1.7Hz,2H),3.90-3.97(m,2H),4.26-4.32(m,2H),4.33-4.39 (m,2H),6.77(s,1H),6.88-6.94(m,1H),7.75(d,J=1.6Hz,1H),7.79-7.84(m,1H),8.07(s,1H);

[0402] MS m / z (ESI): 714.2 [M+H] + .

[0403] The preparation methods of other embodiments are as follows:

[0404]

[0405]

[0406]

[0407]

[0408]

[0409]

[0410]

[0411] two, Biological testing evaluation

[0412] The present invention will be further described and explained below with reference to test examples, but these embodiments are not intended to limit the scope of the present invention.

[0413] Test Example 1: Determination of the inhibitory activity of the compounds of the present invention against EGFR wild-type, EGFR del746-750 / T790M / C797S and EGFR L858R / T790M / C797S mutant kinases.

[0414] Experimental objective: The purpose of this test case is to test the inhibitory activity of the compound against EGFR wild-type, EGFR del746-750 / T790M / C797S and EGFR L858R / T790M / C797S mutant kinases.

[0415] Experimental instruments: Centrifuge (5810R) purchased from Eppendorf, pipettes purchased from Eppendorf or Rainin, and microplate reader purchased from BioTek, USA, model SynergyH1 full-function microplate reader.

[0416] Experimental Methods: This experiment employed the Cisbio HTRF kinase assay (Cisbio #62TK0PEB). The substrate peptide TK and ATP underwent a catalytic reaction in the presence of tyrosine kinases EGFR wild-type, EGFR del746-750 / T790M / C797S, or EGFR L858R / T790M / C797S mutant EGFR. The substrate was phosphorylated, and the kinase activity was characterized by measuring the amount of phosphorylated substrate generated during the reaction. The half-maximal inhibitory concentration (IC50) of the compound against the activity of EGFR wild-type, EGFR del746-750 / T790M / C797S, or EGFR L858R / T790M / C797S mutant kinases was also determined. 50 .

[0417] The specific experimental procedure is as follows:

[0418] The kinase reaction was performed in white 384-well plates (Perkin Elmer #6008280). 1–5 μL of different concentrations of the compound diluted with ddH2O containing 1% DMSO was added to each well. For positive control wells, 1–5 μL of ddH2O containing 1% DMSO was added. Then, 1–5 μL of 0.5–5 nM 4× EGFR wild-type, EGFR del746-750 / T790M / C797S, or EGFR L858R / T790M / C797S mutant kinase solution diluted with Dilution buffer (5× kinase buffer, MgCl2 6.65 mM, MnCl2 1.33 mM, DTT 1.33 mM) was added to each well. For negative control wells, 1–5 μL of Dilution buffer was added. Finally, 1–5 μL of 4 μM kinase solution prepared with 10× Dilution buffer was added to all wells. Add 4× substrate TK solution, and finally add 1-5 μL of 24 μM 4×ATP solution diluted with Dilutionbuffer to start the reaction. After reacting at room temperature for 120 minutes, add 10 μL of detection solution (16 nM TK antibody, 0.5 μM XL665) to each well and react at room temperature in the dark for 20 minutes. Then, detect the chemiluminescence value using a BioTek Synergy H1 microplate reader.

[0419] Experimental data processing methods:

[0420] The percentage inhibition data for the wells treated with the compound were calculated using positive control wells (DMSO control wells) and negative control wells (no kinase added) on a plate: {% inhibition rate = 100 - [(test compound value - negative control value)] / (positive control value - negative control value) × 100}. The IC50 was calculated using a GraphPad Prism approximation to fit different concentrations and corresponding percentage inhibition rate data to a 4-parameter nonlinear logic formula. 50 The values, specifically the data, are shown in the table below:

[0421]

[0422]

[0423] Experimental conclusion:

[0424] Based on the above methods, it can be concluded that the compounds in the embodiments of the present invention have a strong inhibitory effect on the EGFR mutant kinase activity, while the inhibitory effect on the EGFR wild-type kinase activity is relatively small. Comparative data shows that the compounds in the series of embodiments of the present invention have high selectivity in inhibiting the activity of EGFR mutant / wild-type kinase.

[0425] Test Example 2: Determination of the inhibitory activity of the compounds of the present invention against EGFR del746-750 / C797S and EGFR L858R / C797S mutant kinases.

[0426] Experimental objective: The purpose of this test case is to test the inhibitory activity of the compound against EGFR del746-750 / C797S and EGFR L858R / C797S mutant kinases.

[0427] Experimental instruments: Centrifuge (5810R) purchased from Eppendorf, pipettes purchased from Eppendorf or Rainin, and microplate reader purchased from BioTek, USA, model SynergyH1 full-function microplate reader.

[0428] Experimental Methods: This experiment employed the Cisbio HTRF kinase assay (Cisbio #62TK0PEB). The substrate peptide TK and ATP underwent a catalytic reaction in the presence of EGFR del746-750 / C797S or EGFR L858R / C797S mutant tyrosine kinases. The substrate was phosphorylated, and the kinase activity was characterized by measuring the amount of phosphorylated substrate generated during the reaction. The half-maximal inhibitory concentration (IC50) of the compound against the EGFR del746-750 / C797S or EGFR L858R / C797S mutant kinase activity was also determined. 50 .

[0429] The specific experimental procedure is as follows:

[0430] The kinase reaction was performed in white 384-well plates (Perkin Elmer #6008280). 1–5 μL of different concentrations of the compound diluted with ddH2O containing 1% DMSO was added to each well. For positive control wells, 1–5 μL of ddH2O containing 1% DMSO was added. Then, 1–5 μL of 0.5–5 nM 4× EGFR del746-750 / C797S or EGFR L858R / C797S mutant kinase solution diluted with Dilution buffer (5× kinase buffer, MgCl2 6.65 mM, MnCl2 1.33 mM, DTT 1.33 mM) was added to each well. For negative control wells, 1–5 μL of Dilution buffer was added. All wells then contained 1–5 μL of 4 μM 4× substrate TK solution prepared with 10× Dilution buffer. Finally, 1–5 μL of 24 μM Dilution buffer diluted with Dilution buffer was added. The reaction was started with 4×ATP solution. After reacting at room temperature for 120 minutes, 10 μL of detection solution (16 nM TK antibody, 0.5 μM XL665) was added to each well. After reacting at room temperature in the dark for 20 minutes, the chemiluminescence value was detected using a BioTek Synergy H1 microplate reader.

[0431] Experimental data processing methods:

[0432] The percentage inhibition data for the wells treated with the compound were calculated using positive control wells (DMSO control wells) and negative control wells (no kinase added) on a plate: {% inhibition rate = 100 - [(test compound value - negative control value)] / (positive control value - negative control value) × 100}. The IC50 was calculated using a GraphPad Prism approximation to fit different concentrations and corresponding percentage inhibition rate data to a 4-parameter nonlinear logic formula. 50 The values, specifically the data, are shown in the table below:

[0433]

[0434]

[0435] Experimental conclusion:

[0436] Based on the above methods, the compounds in the embodiments of this invention exhibit strong inhibitory effects on the kinase activity of EGFR del746-750 / C797S or EGFR L858R / C797S mutants.

[0437] Test Example 3: Cell Proliferation Inhibition Experiment

[0438] Experimental objective: The purpose of this test case is to test the inhibitory activity of the compound on cell proliferation.

[0439] Experimental instruments: pipettes were purchased from Eppendorf, CO2 incubators were purchased from Thermo Fisher Scientific, USA, and microplate readers were purchased from BioTek, USA, model SynergyH1 full-function microplate readers.

[0440] Experimental Methods: This experiment used CTG (Cell Titer-Glo) chemiluminescence immunoassay to detect the inhibitory activity of the compound on the proliferation of A431 cells and Ba / F3 (EGFR del746-750 / T790M / C797S) cells, and determined the half-maximal inhibitory concentration (IC50) of the compound on cell proliferation. 50 .

[0441] The specific experimental procedure is as follows:

[0442] For A431 cells: On day 1, 90 μL of A431 cell suspension was seeded into 96-well assay plates, with 3000 cells per well. No cells were added to the negative control wells. The plates were incubated overnight at 37°C with 5% CO2. On day 2, 10 μL of serially diluted compound solution was added to each well. For the positive and negative control wells, only 10 μL of DMSO-containing medium was added. The plates were incubated in a CO2 incubator for 72 hours. After 72 hours of incubation, 50 μL of CellTiter Glo was added to each well of the cell plate. The plates were shaken for 2 minutes in the dark and then incubated for 10 minutes. The chemiluminescence value was then measured using a BioTek Synergy H1 microplate reader. The inhibition rate was calculated from the chemiluminescence signal value, and the IC50 of the compound was determined by curve fitting based on the inhibition rate at different concentrations. 50 .

[0443] For Ba / F3 (EGFR del746-750 / T790M / C797S) suspension cells:

[0444] 90 μL of Ba / F3 cell suspension was seeded into each well of a 96-well plate, with 3000 cells per well. No cells were added to the negative control. After standing for 2 hours, 10 μL of serially diluted compound solution was added to each well. 10 μL of culture medium containing DMSO was added to the positive and negative control wells. The plates were incubated in a CO2 incubator for 72 hours and then CTG detection was performed using the same method as for A431 cells.

[0445] Experimental data processing methods:

[0446] The percentage inhibition data for the wells treated with the compound were calculated using positive control wells (DMSO control wells) and negative control wells (no cells) on a plate: {% inhibition rate = 100 - [(test compound value - negative control value)] / (positive control value - negative control value) × 100}. The IC50 was calculated using a GraphPad Prism approximation to fit different concentrations and corresponding percentage inhibition rate data to a 4-parameter nonlinear logic formula. 50 The values, specifically the data, are shown in the table below:

[0447]

[0448]

[0449] Experimental conclusion:

[0450] Based on the above methods, the compounds in the embodiments of this invention showed good inhibitory effects on the proliferation activity of Ba / F3 (EGFR del746-750 / T790M / C797S) mutant cells, while exhibiting weak inhibitory effects on A431 cells. Comparative data show that the compounds in the series of embodiments of this invention have high selectivity in inhibiting the proliferation activity of Ba / F3 (EGFR del746-750 / T790M / C797S) mutant cells.

[0451] Test Example 4: Determination of the inhibitory effect of the compound of the present invention on EGFR phosphorylation in cells.

[0452] Experimental objective: The purpose of this test case is to test the inhibitory activity of the compound on EGFR phosphorylation in cells.

[0453] Experimental apparatus: Microplate shaker (88880024) purchased from Thermo Scientific TM The company purchased the centrifuge (5702R) from Eppendorf, the pipettes from Eppendorf, and the microplate reader from Biotech, USA, model SynergyH1 full-function microplate reader.

[0454] Experimental reagents: Phospho-EGFR (Tyr1068) LANCE Ultra TR-FRET Cellular DetectionKit (Perkin Elmer TRF4016C) contains (5X) LANCE Ultra Lysis Buffer 1, LANCE Ultra Eu-labeled Anti-EGFR (Y1068) Antibody, LANCE Ultra ULight-labeled Anti-EGFR Antibody, EGF (Thermo fisher PHG0311);

[0455] Experimental Methods: This experiment used the Ba / F3 (EGFR del746-750 / T790M / C797S) cell line. The EGFR signaling pathway was activated by EGF stimulation. The inhibitory activity of the compound on the phosphorylation of downstream EGFR (Y1068) was detected, and the half-maximal inhibitory concentration (IC50) of the compound on the EGFR signaling pathway activity was determined. 50 .

[0456] The specific experimental procedure is as follows:

[0457] Seed 3-12 μL of Ba / F3 (EGFR del746-750 / T790M / C797S) cells into 384-well plates, with 100-300 cells per well. Add 2 μL of serially diluted compound solution and incubate at 350 rpm for 2 hours at room temperature. After 2 hours, add 2 μL of EGF to a final concentration of 50 nM and shake at room temperature for 15 minutes. Add 2-5 μL of (5X) LANCE Ultra Lysis Buffer 1 solution and shake at room temperature for 2 hours. After 2 hours, add 5 μL of LANCE Ultra Eu-labeled Anti-EGFR (Y1068) Antibody (PerkinElmer) solution to a final concentration of 0.5 nM and LANCE Ultra U Light-labeled Anti-EGFR Antibody (PerkinElmer) solution to a final concentration of 5 nM, and incubate overnight at room temperature. The fluorescence signal value at 665 nm was measured in each well using a microplate reader. The inhibition rate was calculated from the fluorescence signal value, and the IC50 of the compound was obtained by curve fitting based on the inhibition rate at different concentrations. 50 .

[0458] Experimental data processing methods:

[0459] Percentage inhibition data of wells treated with the test compound is calculated via positive control wells (DMSO control wells) and negative control wells (no cells added) on the plate {% inhibition = 100 - [(test compound value - negative control value)] / (positive control value - negative control value) × 100}. GraphPad Prism is used to fit data of different concentrations and corresponding percentage inhibition rates to a 4-parameter nonlinear logistic equation to calculate IC 50 value.

[0460] Example 1 0.06 Example 2 0.39 Example 6 2.57 Example 7 1.65 Example 9 5.00 Example 10 0.21 Example 16 0.11 Example 18 1.15 Example 20 6.93 Example 25 2.65 Example 29 5.00

[0461] Experimental conclusion:

[0462] It is obtained from the above scheme that the example compounds shown in the present invention have a good inhibitory effect on EGFR phosphorylation in Ba / F3 (EGFR del746-750 / T790M / C797S) cells.

[0463] Test Example 5: Determination of Pharmacokinetics in Balb / C Mice

[0464] 5.1 Research objective:

[0465] Using Balb / C mice as test animals, the pharmacokinetic behavior of the example compound in plasma of mice after oral administration at a dose of 5 mg / kg is studied.

[0466] 5.2 Experimental Protocol

[0467] 5.2.1 Test drug:

[0468] The compound of the example of the present invention, prepared in-house.

[0469] 5.2.2 Test animal:

[0470] Balb / C Mouse (6 animals per example), male, supplied by Shanghai Jiesijie Experimental Animal Co., Ltd., Animal Production License No. (SCXK (Hu) 2013-0006 No. 311620400001794).

[0471] 5.2.3 Formulation prescription:

[0472] 0.5% CMC-Na (1% Tween 80), dissolved by ultrasonication, prepared into a clear solution or uniform suspension.

[0473] 5.2.4 Administration:

[0474] Balb / C mice (6 animals per example), male; after overnight fasting, each mouse is administered orally (p.o.) at a dose of 5 mg / kg, with an administration volume of 10 mL / kg.

[0475] 5.2.5 Sample collection:

[0476] Before and after drug administration, 0.1 mL of blood was collected from the orbital cavity of mice at 0, 0.5, 1, 2, 4, 6, 8 and 24 hours. The blood was placed in EDTA-K2 tubes, centrifuged at 6000 rpm for 6 min at 4℃ to separate the plasma, and stored at -80℃.

[0477] 5.2.6 Sample preparation:

[0478] 1) Add 40 μL of plasma sample to 160 μL of acetonitrile to precipitate, mix, and centrifuge at 3500×g for 5–20 minutes.

[0479] 2) Take 100 μL of the supernatant solution after treatment and analyze the concentration of the analyte by LC / MS / MS.

[0480] 5.2.7 Liquid Chromatography Analysis

[0481] • Liquid phase conditions: Shimadzu LC-20AD pump

[0482] Mass spectrometry conditions: AB Sciex API 4000 mass spectrometer

[0483] • Column: phenomenex Gemiu 5um C18 50×4.6mm

[0484] • Mobile phase: Solution A is a 0.1% formic acid aqueous solution, and solution B is acetonitrile.

[0485] • Flow rate: 0.8 mL / min

[0486] • Elution time: 0-4.0 minutes, eluent as follows:

[0487]

[0488]

[0489] 5.3 Experimental Results and Analysis

[0490] The main pharmacokinetic parameters were calculated using WinNonlin 6.1. The results of the mouse pharmacokinetic experiment are shown in the table below:

[0491] Example 1-FA 2.0 1933 11593 11631 2.9 4.7 Example 2-FA 2.0 2113 21017 21383 4.0 6.3 Example 6-FA 4.0 3233 38816 38967 2.8 6.1 Example 7-FA 2.0 2707 25304 25344 2.5 5.3 Example 9-FA 2.0 1680 16368 16511 5.6 6.0 Example 10-FA 2.0 2027 25171 25328 3.2 6.1 Example 11-FA 2.0 1340 11759 11844 4.2 5.6 Example 15-FA 2.0 2280 8168 8178 1.1 3.0 Example 26-FA 2.0 1470 12341 12535 4.2 6.0 Example 29-FA 2.0 2323 18009 18175 3.5 5.5 Example 30-FA 2.0 3327 20587 20594 1.8 3.8

[0492] Note: FA stands for the formate of the corresponding compound.

[0493] Experimental conclusion:

[0494] As can be seen from the results of the mouse pharmacokinetic experiments in the table, the compounds in the embodiments of the present invention exhibit good metabolic properties, with low exposure AUC and high maximum plasma concentration C. max They all performed well.

[0495] Test Example 6: In vivo efficacy test of the compounds in the embodiments of the present invention

[0496] 6.1 Experimental Objective

[0497] Compounds with significant efficacy and low toxicity were screened through in vivo pharmacodynamic experiments.

[0498] 6.2 Main Instruments and Materials for the Experiment

[0499] 6.2.1 Instruments:

[0500]

[0501]

[0502] 6.2.2 Reagents:

[0503] Matrigel Corning 354234 FBS Gibco 10099-141C Trypsin Gibco 25200-072 RPMI1640 Hyclone SH30809.01 Puromycin Gibco A11138-03 HPMC Sigma H3785

[0504] 6.2.3 Animals:

[0505] NOD / SCID mice, 6-8 weeks old, female, purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.

[0506] 6.3 Experimental Procedure

[0507] 6.3.1 Cell Culture

[0508] PC9 (EGFR Del19 / T790M / C797S) cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum. PC9 (EGFR Del19 / T790M / C797S) cells in the exponential growth phase were collected.

[0509] 6.3.2 Cell Seeding

[0510] The experimental mice were injected subcutaneously on the right back (subcutaneous tissue near the forelimb on the right side of the mouse) with 1×10 7 PC9 (EGFRDel19 / T790M / C797S) cells were resuspended in a 1:1 mixture of PBS and matrix gel (0.1 ml / cell). Tumor growth was observed regularly, and the day of tumor cell inoculation was defined as day 0.

[0511] 6.3.3 Tumor measurement, grouping, and drug administration in tumor-bearing mice

[0512] a. On day 7, tumor volume data were measured, and tumor volumes between 100-200 mm were selected. 3 Mice within the range, with an average volume of 140 mm² 3 Mice were randomly grouped and given medication based on tumor size and body weight.

[0513] c. Based on the grouping results, begin administering the test drug (administration route: oral administration; administration volume: 10 mL / kg; administration frequency: once a day; administration period: 21 days; solvent: 0.5% HPMC).

[0514] d. After starting the test drug, the tumor was measured and weighed twice a week.

[0515] e. Euthanize the animals after the experiment.

[0516] f. Process the data using software such as Excel. Calculation of the tumor inhibition rate (TGI%) of the compound: TGI% = [1 - (T... i -T0) / (C i -C0)]×100%; where, T i Let T0 be the tumor volume on day i in the treatment group, and C be the tumor volume on the day of grouping in the treatment group. i Ci represents the tumor volume of the solvent control group on day i, and C0 represents the tumor volume of the solvent control group on the day of grouping.

[0517] 6.4 The test data are shown in the table below:

[0518]

[0519]

[0520] 6.5 Experimental Results

[0521] The results above show that the compounds in this patent have a good tumor inhibition rate and good safety.

[0522] III. Example 1: Study on the free alkali crystal form

[0523] 1. Experimental apparatus

[0524] 1.1 Some parameters of physicochemical testing instruments

[0525]

[0526] 1.2 Instruments and Liquid Chromatography Analysis Conditions

[0527] 1.2.1 Instruments and Equipment

[0528]

[0529]

[0530] 1.2.2 Chromatographic conditions

[0531] Chromatographic column: ZORBAX (SB-C8, 3.5μm, 4.6*75mm)

[0532] Flow rate: 1 mL / min

[0533] Column temperature: 40℃

[0534] Detection wavelength: 235nm

[0535] Injection volume: 5.0 μL

[0536] Running time: 15min

[0537] Diluent: ACN-water (v / v, 1:1)

[0538] Mobile phase: A: Water (0.05% trifluoroacetic acid); B: Acetonitrile (0.05% trifluoroacetic acid)

[0539] 0.00 90 10 3.00 70 30 10.00 50 50 12.00 30 70 12.01 90 10 15.00 90 10

[0540] 2. Preparation of crystal form

[0541] 1. Preparation of crystal form A

[0542] Approximately 20 mg of free amorphous base was weighed into a 2 mL glass bottle, and 200 μL of organic solvent (e.g., EtOH, Acetone, ACN, THF, EA, Toluene, IPAC, MEK, 2-Me-THF, 1,4-Dioxane, 3-pentanone, and IPA) was added. The mixture was stirred at 50°C for 3 days. The solid was then rapidly centrifuged, the supernatant was removed, and the solid was vacuum dried at 40°C to obtain crystal form A. Analysis revealed the following properties: Figure 1 The XRPD diagram shown is as follows: Figure 2 The DSC diagram shown and as follows Figure 3 The TGA diagram shown.

[0543] 2. Preparation of crystal form B

[0544] Approximately 400 mg of free amorphous base was weighed and dissolved in 2 mL of methanol at 50 °C upon heating. Then, 0.6 mL of methyl tert-butyl ether was added at 50 °C and stirred for a certain period, resulting in the precipitation of a large amount of white solid. The mixture was stirred at 50 °C for 2 hours, and then 0.4 mL of methyl tert-butyl ether was added. After cooling and stirring, the mixture was filtered. The filter cake was washed with methyl tert-butyl ether and dried under vacuum at 40 °C to constant weight, yielding crystal form B. Analysis revealed the following properties: Figure 4 The XRPD diagram shown and as follows Figure 5 The DSC diagram shown is shown.

[0545] 3. Preparation of crystal form C

[0546] Approximately 20 mg of free amorphous alkali was weighed into a 2 mL glass bottle, 200 μL of water was added, and the mixture was stirred at 50 °C for 3 days. The solid was then rapidly centrifuged, the supernatant was removed, and the solid was vacuum dried at 40 °C to obtain crystalline form C. Analysis revealed the following properties: Figure 6 The XRPD diagram shown and as follows Figure 7 The DSC diagram shown.

[0547] 4. Preparation of crystal form D

[0548] Approximately 50 mg of free amorphous base was weighed and dissolved in 0.5 mL of methanol at 50 °C upon heating. Then, 1.5 mL of isopropyl ether was added at 50 °C, causing turbidity and the formation of an oil. After stirring for a certain period, a large amount of white solid precipitated. The mixture was stirred at 50 °C for 2 hours, followed by the addition of 2 mL of isopropyl ether. The mixture was then cooled, stirred, and filtered. The filter cake was washed with isopropyl ether and dried under vacuum at 40 °C to constant weight, yielding crystal form D. Analysis revealed the following properties: Figure 8 The XRPD diagram shown and as follows Figure 9 The DSC diagram shown.

[0549] 5. Preparation of crystal form E

[0550] Approximately 20 mg of free amorphous alkali was weighed into a 2 mL glass bottle, and 200 μL of 88% Acetone was added. The mixture was stirred at 50 °C for 3 days. The solid was then rapidly centrifuged, the supernatant was removed, and the solid was vacuum dried at 40 °C to obtain crystal form E. Analysis revealed the following properties: Figure 10 The XRPD diagram shown and as follows Figure 11 The DSC diagram shown.

[0551] 6. Preparation of crystal form F

[0552] Approximately 10 mg of free base amorphous form was weighed and dissolved in 200 μL of DCM at room temperature with stirring. Then, 200 μL of n-heptane was added at room temperature, causing turbidity to form an oil. After stirring for a certain period, a large amount of white solid precipitated. Another 400 μL of n-heptane was added and stirred for 2 hours. The solid was then rapidly centrifuged, the supernatant was removed, and the solid was dried under vacuum at 40°C to obtain crystal form F. Analysis revealed the following properties: Figure 12 The XRPD diagram shown and as follows Figure 13 The DSC diagram shown.

[0553] 7. Preparation of crystal form G

[0554] Approximately 10 mg of free amorphous base was weighed and added to 200 μL of 88% Acetone. The solution was heated to 70°C and immediately stirred at 0°C. A large amount of white solid precipitated. The solid was rapidly centrifuged, the supernatant was removed, and the solution was dried under vacuum at 40°C to obtain crystal form G. Analysis revealed the following properties: Figure 14 The XRPD diagram shown and as follows Figure 15 The DSC diagram shown.

[0555] 8. Preparation of crystal form J

[0556] Approximately 20 mg of the amorphous crude free alkali was weighed into a 2 mL glass bottle, and 100 μL of organic solvent (MTBE or IPA are acceptable options) was added. The mixture was stirred at room temperature for 12 hours. Finally, the solid was rapidly centrifuged, the supernatant was removed, and the solid was dried under vacuum at 40 °C to obtain crystal form J. Analysis revealed the following properties: Figure 16 The XRPD diagram shown and as follows Figure 17 The DSC diagram shown.

[0557] 9. Preparation of crystal form K

[0558] Approximately 20 mg of the free alkali crude product (amorphous form) was weighed into a 2 mL glass bottle, 100 μL of water was added, and the mixture was stirred at room temperature for 12 h. Finally, the solid was rapidly centrifuged, the supernatant was removed, and the product was vacuum dried at 40 °C to obtain crystalline form K. Analysis revealed the following properties: Figure 18 The XRPD diagram shown and as follows Figure 19 The DSC diagram shown.

[0559] 10. Preparation of crystal form L

[0560] Approximately 10 mg of free alkali crystal form A was weighed and added to 100 μL of DCM, stirred at room temperature until dissolved. Then, 100 μL of isopropyl ether was added at room temperature, causing turbidity to form an oil. After continued stirring for a certain period, a large amount of white solid precipitated. After stirring at room temperature for 12 hours, the solid was rapidly centrifuged, the supernatant was removed, and the solid was dried under vacuum at 40°C to obtain crystal form L. Analysis revealed the following properties: Figure 20 The XRPD diagram shown.

[0561] 11. Preparation of crystal form M

[0562] Approximately 10 mg of free alkali crystalline form C was weighed into a 2 mL glass bottle, 100 μL of DCM was added, and the mixture was stirred at room temperature for 12 h. Finally, the solid was rapidly centrifuged, the supernatant was removed, and the solid was dried under vacuum at 40 °C to obtain crystalline form M. Analysis revealed the following properties: Figure 21 The XRPD diagram shown.

[0563] 3. Solid stability test

[0564] 3.1 Experimental Objective:

[0565] The physicochemical stability of free base crystal forms A, B, C, and D at a high temperature of 60°C was investigated to provide a basis for crystal form screening and compound storage.

[0566] 3.2 Experimental Procedure:

[0567] Take approximately 2 mg each of free alkali crystal form A, crystal form B, crystal form C, and crystal form D, and place them in an oven at 60℃ for 5 and 10 days. Determine the content using HPLC with external standard method, and calculate the changes in related substances using the chromatographic peak area normalization method.

[0568] 3.3 Experimental Results:

[0569] Physicochemical stability results of free bases in different crystal forms:

[0570]

[0571]

[0572] 4. Dynamic hygroscopicity test

[0573] 4.1 Experimental Objective

[0574] The hygroscopicity of the free base crystal form C of compound was investigated under different relative humidity conditions, providing a basis for the screening and storage of compound crystal forms.

[0575] 4.2 Experimental Procedure:

[0576] The free alkali crystal form C of the compound was placed in saturated water vapor with different relative humidities to allow the compound to reach dynamic equilibrium with the water vapor, and the percentage of the compound's weight gain due to moisture absorption after equilibrium was calculated.

[0577] 4.3 Experimental Results:

[0578] The free alkali crystal form C gains approximately 8.263% weight upon moisture absorption at 80% RH, indicating strong hygroscopicity. After one cycle of moisture absorption and desorption at 0-95% relative humidity, the XRPD spectrum of the free alkali crystal form C remained unchanged, indicating that the crystal form did not transform.

[0579] 5. Solubility experiments in different media

[0580] 5.1 Experimental Objective

[0581] This study compares the solubility of free basal crystal forms A and C in water, simulated gastric juice (SGF), fasted simulated intestinal juice (FaSSIF), and non-fasted simulated intestinal juice (FeSSIF), as well as their solubility in buffer solutions ranging from pH 1 to pH 8. This provides a basis for druggability assessment.

[0582] 5.2 Experimental Procedure:

[0583] Approximately 2 mg of free base crystal forms A and C were suspended in different media for 2 hours, and the thermodynamic solubility of the compounds at 37 °C was determined by HPLC using the external standard method.

[0584] 5.3 Experimental Results: As shown in Table 15

[0585] Table 15

[0586]

[0587]

[0588] 6. Animal PK Studies

[0589] 6.1 Experimental Objective:

[0590] Using SD rats as test animals, this study investigated the pharmacokinetic behavior of a single oral administration of free basal crystalline form A in rats (plasma) and compared changes in exposure levels. The study also investigated the pharmacokinetics of a single intravenous administration of free basal crystalline form A and calculated its bioavailability after oral administration.

[0591] 6.2 Experimental Procedure:

[0592] Free alkali crystal form A was suspended in an aqueous solution containing 0.5% HPMC K4M and administered to rats by gavage. Three parallel rats were administered the drug at a dose of free alkali crystal form A (30 mg / kg clear solution). Free alkali crystal form A was dissolved in a physiological saline solution containing 20% ​​HP-β-CD, filtered through a filter membrane, and then injected into rats. Three parallel rats were administered the drug.

[0593] 6.3 Experimental Results:

[0594] Parameters Crystal form A <![CDATA[t max (h)]]> 4.0 <![CDATA[C max (ng / mL)]]> 1454.7 <![CDATA[AUC 0-24 (ng / mL*h)]]> 17577.2 <![CDATA[AUC 0-∞ (ng / mL*h)]]> 22529.0 <![CDATA[t 1 / 2 (h)]]> 11.1 <![CDATA[MRT 0-∞ (h)]]> 8.1 F(％) 20.1 FormμLation 0.5% HPMC

[0595] Rat IV Dose: 2mg / kg

[0596]

[0597]

[0598] 7. Stable crystal form confirmation experiment

[0599] 7.1 Experimental Objective:

[0600] A relatively stable compound crystal form was found through crystal form slurry experiments, crystal form competition experiments, and stability investigation experiments.

[0601] 7.2 Experimental Procedure:

[0602] Select an organic solvent and water with a certain solubility, suspend different crystal forms in the solvent system, and stir and slurry them at a certain temperature; place different crystal forms under different conditions to observe the changes in crystal form. Finally, process the solid, measure the XRPD of the solid, and compare the results.

[0603] 7.3 Experimental Results:

[0604] Transformation relationships of different crystal forms in solvents:

[0605] MeOH Crystal form B was extracted by dissolution. Dissolve EtOH It remains in crystal form A after 3 days at 50℃. It transforms into crystal form A at 50℃ for 3 days. Acetone It remains in crystal form A after 3 days at 50℃. It transforms into crystal form A at 50℃ for 3 days. ACN It remains in crystal form A after 3 days at 50℃. It transforms into crystal form A at 50℃ for 3 days. THF It remains in crystal form A after 3 days at 50℃. It remains in crystal form B after 3 days at 50℃. EA It remains in crystal form A after 3 days at 50℃. It remains in crystal form B after 3 days at 50℃. H2O Transformed into crystal form C Transformed into crystal form C

[0606] Transformation relationships of different crystal forms under different influencing factors:

[0607] Crystal form A One day to transform into crystal form C No change after 5 days, peak of crystal form C appears after 10 days. Crystal form D Transformation into crystal form B -

[0608] In nonsolvents, free alkali crystal form A is a thermodynamically stable crystal form.

[0609] IV. Example 1: Study on Salt-Type Crystal Form

[0610] 1. Screening of compound salt crystal forms

[0611] 1.1 Screening of compound salt types

[0612] 1.1.1 Experimental Objective:

[0613] By selecting different counterionic acids and using appropriate crystallization methods, we can detect which counterionic acids can form compound salts.

[0614] 1.1.2 Experimental Procedure:

[0615] 1) Instruments and equipment

[0616] Analytical balance BSA224S-CW Sartorius Ultrasonic cleaner SK5200LHC Shanghai Kedao Ultrasonic Instruments pipette Eppendorf (50 mL, 1000 μL) Eppendorf

[0617] 2) Operating Procedures

[0618] 1.2.1 Natural evaporation method using methanol as solvent

[0619] Weigh 300 mg of free base and dissolve it completely in 8 mL of methanol at room temperature. Divide the methanol solution of free base into 20 equal portions. Add a certain amount of acid to each portion (base:acid = 1:1.2 molar ratio). Then, allow the solvent to evaporate at room temperature. After evaporation, the sample properties are not good. Add 200 μL of EA to each portion and pulverize. Details are as follows:

[0620] 1 1M hydrochloric acid in MeOH Still clarifying amorphous Salt 2 1M oxalic acid in MeOH Still clarifying It contains crystals, but has poor crystallinity. Free base 3 0.5M tartaric acid in MeOH Still clarifying Oil Free base 4 0.25M fumaric acid in EtOH Still clarifying Oil Free base 5 1M maleic acid in MeOH Still clarifying It contains crystals, but has poor crystallinity. Free base 6 1M formic acid in MeOH Still clarifying Very small amount of crystals Free base 7 1M acetic acid in MeOH Still clarifying Basically the same as 10XRD Free base 8 1M succinic acid in MeOH Still clarifying It contains crystals, but has poor crystallinity. Free base 9 1M adipic acid in MeOH Still clarifying Almost invisible solid Free base 10 1M malic acid in MeOH Still clarifying Good crystallinity Free base 11 1M benzoic acid in MeOH Still clarifying amorphous Free base 12 MeOH Still clarifying amorphous Free base

[0621] Results and Discussion: The above experiments yielded hydrochloride, phosphate, and a free alkali crystal form.

[0622] 1.2.2 Solid-liquid reaction crystallization using Acetone as solvent

[0623] Weigh 20 mg of free base, add 0.2 mL of Lacetone, stir at room temperature to form a white suspension, then add acid to the suspension (base:acid = 1:1.2 molar ratio) to carry out the reaction, as follows:

[0624] 1 1M phosphoric acid in MeOH Undissolved water turns pale yellow. 50℃ compared to 2h It may turn into salt.

[0625] Results and Discussion: Phosphate was obtained from the above experiments.

[0626] 1.2.3 Using DCM as the solvent for liquid-liquid reaction crystallization

[0627] Weigh 10 mg of free base, add 0.1 mL of LCM, stir at room temperature until dissolved, then add acid to the solution (base:acid = 1:1.2 molar ratio) to carry out the reaction, as follows:

[0628] 1 1M phosphoric acid in MeOH Immediately cloudy phosphate

[0629] Results and Discussion: Phosphate was obtained from the above experiments.

[0630] 1.2.4 Solid-liquid reaction crystallization using EA as solvent

[0631] Weigh 10 mg of free base, add 0.1 mL of EA, stir and suspend at 50 °C, then add acid (base:acid = 1:1.2 molar ratio) to the suspension to carry out the reaction, as follows:

[0632] 1 1M phosphoric acid in MeOH Localized areas turn pale yellow phosphate

[0633] 1.2.5 Using MeOH as solvent, liquid-liquid reaction is coupled with crystallization.

[0634] Weigh 10 mg of free base, add 0.1 mL of MeOH, stir at 50 °C until dissolved, then add acid (base:acid = 1:1.2 molar ratio) to the solution to carry out the reaction, as follows:

[0635] 1 1M phosphoric acid in MeOH After dissolving, stirring will cause precipitation. phosphate

[0636] Results and Discussion: Phosphate was obtained from the above experiments.

[0637] 1.2.6 Natural evaporation method using DCM as solvent

[0638] Weigh 194 mg of free base and dissolve it completely in 6.5 mL of DCM at room temperature. Divide the methanol solution of free base into 13 equal portions. Add a certain amount of acid to each portion (base:acid = 1:1.2 molar ratio). Then, allow the solvent to evaporate at room temperature in an open container. After evaporation, an oil is formed. Add 200 μL of each oil to each portion and slurry. Details are as follows:

[0639] 1 1M mesylate in MeOH Still clarifying Oil There is a solid, methanesulfonate.

[0640] Results and Discussion: The above experiments screened out methanesulfonates.

[0641] 1.2.7 Solid-liquid reaction crystallization using 2-Me-THF as solvent

[0642] Weigh 10 mg of free base, add 0.1 mL of 2-Me-THF, stir the white suspension at room temperature, and then add acid (base:acid = 1:1.2 molar ratio) to the suspension to carry out the reaction, as follows:

[0643]

[0644]

[0645] Results and Discussion: The above experiments yielded methanesulfonate.

[0646] 2.1.3 Experimental Results

[0647] Through salt type screening experiments, phosphate, hydrochloride, methanesulfonate and their crystal forms were obtained, and the crystal form was named crystal form A.

[0648] 2. Hygroscopicity test

[0649] 2.1 Experimental Objective

[0650] The hygroscopicity of different salts of compounds under different relative humidity conditions was investigated to provide a basis for the screening and storage of compound salts.

[0651] 2.2 Experimental Scheme:

[0652] The compound salt was placed in saturated water vapor at different relative humidities to allow the compound to reach dynamic equilibrium with the water vapor, and the percentage of the compound's weight gain due to moisture absorption after equilibrium was calculated.

[0653] 2.3 Experimental Results:

[0654] 2.3.1 Hygroscopicity of salts of compound formula (V)

[0655] 1) Phosphate crystal form A gains 8.31% weight by absorbing moisture under RH 80% conditions, indicating strong hygroscopicity. After one cycle of moisture absorption and desorption under 0-95% relative humidity, the XRPD spectrum of phosphate crystal form A did not change, meaning the crystal form did not change.

[0656] 2) Hydrochloride crystal form A gains 15.31% weight upon moisture absorption under RH 80% conditions, exhibiting extremely strong hygroscopicity. Furthermore, after one cycle of moisture absorption and desorption under 0-95% relative humidity conditions, the XRPD spectrum of hydrochloride crystal form A changes, indicating a crystal form transformation.

[0657] 3) Methanesulfonate crystal form A has a moisture absorption weight gain of 18.36% under RH 80% conditions, and is extremely hygroscopic.

[0658] 2.4 Experimental Conclusions

[0659] Phosphate crystal form A has the lowest hygroscopicity.

[0660] 3. Solid stability test

[0661] 3.1 Experimental Objective:

[0662] The physicochemical stability of the compounds in free alkali crystal form A, phosphate crystal form A, and hydrochloride crystal form A at a high temperature of 60°C was investigated to provide a basis for salt form screening and compound salt storage.

[0663] 3.2 Experimental Procedure:

[0664] Take about 2 mg of free alkali or different salts and place them in an oven at 60°C for 5 and 10 days. Determine the salt content by HPLC using the external standard method and calculate the changes in salt-related substances by the chromatographic peak area normalization method.

[0665] 3.3 Experimental Results:

[0666]

Claims

1. A crystal form of the compound shown in formula (I), characterized in that, ， The crystal forms are AG and JM, wherein: The X-ray powder diffraction pattern of crystal form A shows diffraction peaks at 2θ of 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, and 20.9±0.2°. The X-ray powder diffraction pattern of crystal form B shows diffraction peaks at 2θ of 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, and 22.5±0.2°. The X-ray powder diffraction pattern of crystal form C shows diffraction peaks at 2θ of 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 5.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, and 17.7±0.2°. The X-ray powder diffraction pattern of crystal form D shows diffraction peaks at 2θ of 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 17.9±0.2°, and 15.0±0.2°. The X-ray powder diffraction pattern of crystal form E shows diffraction peaks at 2θ of 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, and 21.5±0.2°. The X-ray powder diffraction pattern of crystal form F shows diffraction peaks at 2θ of 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, and 22.5±0.2°. The X-ray powder diffraction pattern of crystal form G shows diffraction peaks at 2θ of 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, and 23.5±0.2°. The X-ray powder diffraction pattern of crystal form J shows diffraction peaks at 2θ of 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, and 16.8±0.2°. The X-ray powder diffraction pattern of crystal form K shows diffraction peaks at 2θ of 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, and 10.0±0.2°. The X-ray powder diffraction pattern of crystal form L shows diffraction peaks at 2θ of 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, and 15.3±0.2°. The X-ray powder diffraction pattern of crystal form M has diffraction peaks at 2θ of 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, and 21.9±0.2°.

2. The crystal form of the compound according to claim 1, characterized in that, The X-ray powder diffraction pattern of crystal form A may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, 20.9±0.2°, 24.4±0.2°, and 18.9±0.2°. The X-ray powder diffraction pattern of crystal form B may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, 22.5±0.2°, 8.1±0.2°, and 15.7±0.2°. The X-ray powder diffraction pattern of crystal form C may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, 17.7±0.2°, 22.3±0.2°, and 21.0±0.2°. The X-ray powder diffraction pattern of the crystal form D may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.0±0.2°, and 8.1±0.2°. The X-ray powder diffraction pattern of crystal form E may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, 21.5±0.2°, 5.2±0.2°, and 14.1±0.2°. The X-ray powder diffraction pattern of the crystal form F may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, 22.5±0.2°, 18.8±0.2°, and 15.5±0.2°. The X-ray powder diffraction pattern of the crystal form G may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, 23.5±0.2°, 21.3±0.2°, and 28.0±0.2°. The X-ray powder diffraction pattern of crystal form J may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, 16.8±0.2°, 21.3±0.2°, and 25.4±0.2°. The X-ray powder diffraction pattern of the crystal form K may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, 10.0±0.2°, 7.8±0.2°, and 11.6±0.2°. The X-ray powder diffraction pattern of the crystal form L may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, 15.3±0.2°, 24.6±0.2°, and 18.5±0.2°. The X-ray powder diffraction pattern of the crystal form M may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, 21.9±0.2°, 11.9±0.2°, and 27.7±0.2°.

3. The crystal form of the compound according to claim 2, characterized in that, The X-ray powder diffraction pattern of crystal form A may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, 20.9±0.2°, 24.4±0.2°, and 18.9±0.2°. The X-ray powder diffraction pattern of crystal form B may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, 22.5±0.2°, 8.1±0.2°, and 15.7±0.2°. The X-ray powder diffraction pattern of crystal form C may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, 17.7±0.2°, 22.3±0.2°, and 21.0±0.2°. The X-ray powder diffraction pattern of crystal form D may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.0±0.2°, and 8.1±0.2°. The X-ray powder diffraction pattern of crystal form E may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, 21.5±0.2°, 5.2±0.2°, and 14.1±0.2°. The X-ray powder diffraction pattern of the crystal form F may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, 22.5±0.2°, 18.8±0.2°, and 15.5±0.2°. The X-ray powder diffraction pattern of the crystal form G may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, 23.5±0.2°, 21.3±0.2°, and 28.0±0.2°. The X-ray powder diffraction pattern of crystal form J may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, 16.8±0.2°, 21.3±0.2°, and 25.4±0.2°. The X-ray powder diffraction pattern of the crystal form K may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, 10.0±0.2°, 7.8±0.2°, and 11.6±0.2°. The X-ray powder diffraction pattern of the crystal form L may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, 15.3±0.2°, 24.6±0.2°, and 18.5±0.2°. The X-ray powder diffraction pattern of the crystal form M may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, 21.9±0.2°, 11.9±0.2°, and 27.7±0.2°.

4. The crystal form of the compound according to claim 1, characterized in that, The X-ray powder diffraction pattern of crystal form A includes 10 diffraction peaks at any of the following 2θ values: 17.7±0.2°, 21.5±0.2°, 23.9±0.2°, 14.7±0.2°, 7.7±0.2°, 12.3±0.2°, 15.2±0.2°, 20.9±0.2°, 24.4±0.2°, 18.9±0.2°, 30.8±0.2°, 23.5±0.2°, 13.0±0.2°, 16.9±0.2°, and 17.3±0.2°. The X-ray powder diffraction pattern of crystal form B includes 10 diffraction peaks located at 2θ values ​​of 22.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.2±0.2°, 19.4±0.2°, 12.4±0.2°, 26.0±0.2°, 22.5±0.2°, 8.1±0.2°, 15.7±0.2°, 21.3±0.2°, 14.6±0.2°, 29.4±0.2°, 32.0±0.2°, and 32.9±0.2°. The X-ray powder diffraction pattern of crystal form C includes 10 diffraction peaks located at 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 9.2±0.2°, 25.3±0.2°, 4.8±0.2°, 14.1±0.2°, 21.5±0.2°, 17.7±0.2°, 22.3±0.2°, 21.0±0.2°, 20.2±0.2°, 23.1±0.2°, 28.5±0.2°, 18.0±0.2°, and 19.9±0.2°. The X-ray powder diffraction pattern of crystal form D includes 10 diffraction peaks located at 2θ values ​​of 23.3±0.2°, 19.8±0.2°, 16.9±0.2°, 22.5±0.2°, 14.7±0.2°, 24.9±0.2°, 20.6±0.2°, 17.9±0.2°, 15.0±0.2°, 8.1±0.2°, 15.6±0.2°, 7.6±0.2°, 31.8±0.2°, 19.4±0.2°, and 21.2±0.2°. The X-ray powder diffraction pattern of crystal form E includes 10 diffraction peaks located at 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 4.8±0.2°, 24.6±0.2°, 9.1±0.2°, 22.2±0.2°, 25.2±0.2°, 21.5±0.2°, 5.2±0.2°, 14.1±0.2°, 20.9±0.2°, 15.2±0.2°, 18.6±0.2°, 17.6±0.2°, and 28.5±0.2°. The X-ray powder diffraction pattern of the crystal form F includes 10 diffraction peaks selected from 2θ values ​​of 5.0±0.2°, 16.2±0.2°, 4.8±0.2°, 19.8±0.2°, 14.8±0.2°, 18.3±0.2°, 22.1±0.2°, 22.5±0.2°, 18.8±0.2°, 15.5±0.2°, 23.5±0.2°, 9.1±0.2°, 25.0±0.2°, 19.0±0.2°, and 26.5±0.2°. The X-ray powder diffraction pattern of the crystal form G includes 10 diffraction peaks located at 2θ values ​​of 19.0±0.2°, 26.5±0.2°, 9.1±0.2°, 25.0±0.2°, 4.7±0.2°, 14.0±0.2°, 17.5±0.2°, 23.5±0.2°, 21.3±0.2°, 28.0±0.2°, 22.2±0.2°, 20.7±0.2°, 24.2±0.2°, 23.1±0.2°, and 8.8±0.2°. The X-ray powder diffraction pattern of crystal form J includes 10 diffraction peaks selected from 2θ values ​​of 19.2±0.2°, 14.1±0.2°, 9.8±0.2°, 22.4±0.2°, 23.6±0.2°, 15.9±0.2°, 11.2±0.2°, 16.8±0.2°, 21.3±0.2°, 25.4±0.2°, 18.7±0.2°, 8.5±0.2°, 28.4±0.2°, 19.6±0.2°, and 20.4±0.2°. The X-ray powder diffraction pattern of the crystal form K includes 10 diffraction peaks selected from 2θ values ​​of 25.2±0.2°, 16.9±0.2°, 20.5±0.2°, 18.5±0.2°, 23.8±0.2°, 20.8±0.2°, 10.2±0.2°, 10.0±0.2°, 7.8±0.2°, 11.6±0.2°, 24.2±0.2°, 21.8±0.2°, 19.7±0.2°, 19.1±0.2°, and 15.4±0.2°. The X-ray powder diffraction pattern of the crystal form L includes 10 diffraction peaks selected from 2θ values ​​of 15.9±0.2°, 19.6±0.2°, 23.2±0.2°, 18.1±0.2°, 22.3±0.2°, 4.8±0.2°, 21.7±0.2°, 15.3±0.2°, 24.6±0.2°, 18.5±0.2°, 20.0±0.2°, 18.8±0.2°, 26.3±0.2°, 9.0±0.2°, and 17.2±0.2°. The X-ray powder diffraction pattern of the crystal form M contains 10 diffraction peaks at any of the following 2θ values: 18.8±0.2°, 26.3±0.2°, 8.9±0.2°, 4.5±0.2°, 13.7±0.2°, 24.7±0.2°, 17.2±0.2°, 21.9±0.2°, 11.9±0.2°, 27.7±0.2°, 24.1±0.2°, 21.0±0.2°, 22.9±0.2°, 13.2±0.2°, and 8.5±0.2°.

5. The crystal form of the compound according to claim 1, characterized in that, The X-ray powder diffraction pattern of crystal form A is shown in Figure 1, the DSC pattern is shown in Figure 2, and the TGA pattern is shown in Figure 3. The X-ray powder diffraction pattern of crystal form B is shown in Figure 4, and the DSC pattern is shown in Figure 5. The X-ray powder diffraction pattern of crystal form C is shown in Figure 6, and the DSC pattern is shown in Figure 7. The X-ray powder diffraction pattern of the crystal form D is shown in Figure 8, and the DSC pattern is shown in Figure 9. The X-ray powder diffraction pattern of the crystal form E is shown in Figure 10, and the DSC pattern is shown in Figure 11. The X-ray powder diffraction pattern of the crystal form F is shown in Figure 12, and the DSC pattern is shown in Figure 13. The X-ray powder diffraction pattern of the crystal form G is shown in Figure 14, and the DSC pattern is shown in Figure 15. The X-ray powder diffraction pattern of crystal form J is shown in Figure 16, and the DSC pattern is shown in Figure 17. The X-ray powder diffraction pattern of the crystal form K is shown in Figure 18, and the DSC pattern is shown in Figure 19. The X-ray powder diffraction pattern of the crystal form L is shown in Figure 20. The X-ray powder diffraction pattern of the crystal form M is shown in Figure 21.

6. The crystal form of the compound according to claim 5, characterized in that, The positions of the top ten diffraction peaks with the highest relative peak intensities in the X-ray powder diffraction patterns of crystal forms A, B, C, D, E, F, G, J, K, L, and M have a 2θ error of ±0.2° compared to the corresponding diffraction peaks in Figures 1, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 21.

7. A method for preparing the crystal form of the compound according to any one of claims 1-6, wherein the method is method one, method two, or method three. Method 1 specifically includes the following steps: 1) Weigh an appropriate amount of the free base of compound I and suspend it in a poor solvent; 2) Shake the suspension; 3) The suspension was rapidly centrifuged to remove the supernatant, and the remaining solid was dried to obtain the target product; in: The undesirable solvent is selected from acetone, ethyl acetate, isopropyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, 1,4-dioxane, toluene, isopropanol, 2-butanone, 3-pentanone, methyl tert-butyl ether, or water. Method 2 specifically includes the following steps: 1) Weigh an appropriate amount of the free base of compound I and dissolve it in a good solvent; 2) Optionally, add a poor solvent to the resulting solution and stir until a solid precipitates. 3) Optionally, add a poor solvent to the obtained solution, centrifuge rapidly, remove the supernatant, and dry the remaining solid to obtain the target product; in: The benign solvent is selected from methanol, dichloromethane, or tetrahydrofuran; The unsuitable solvents are selected from heptane, water, methyl tert-butyl ether, toluene, or isopropyl ether; Method 3 specifically includes the following steps: 1) Weigh an appropriate amount of the free base of compound I and dissolve it by heating with a good solvent; 2) Quickly place the resulting solution at a low temperature and stir until a solid precipitates; 3) The obtained suspension was rapidly centrifuged to remove the supernatant, and the remaining solid was dried to obtain the target product; in: The benign solvent is 88% acetone.

8. An acid salt of the compound shown in formula (I), characterized in that, ， in: The acid in the acid salt is selected from hydrochloric acid, phosphoric acid, or methanesulfonic acid.

9. The acid salt according to claim 8, characterized in that, It is the acid salt crystal form of the compound shown in formula (I); the acid salt crystal form is a phosphate crystal form, a hydrochloride crystal form, or a methanesulfonate crystal form; The X-ray powder diffraction pattern of the phosphate crystal form A shows diffraction peaks at 2θ of 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, and 21.4±0.2°. The X-ray powder diffraction pattern of the hydrochloride crystal form A shows diffraction peaks at 2θ of 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, and 29.1±0.2°. The X-ray powder diffraction pattern of the methanesulfonate crystal form A has diffraction peaks at 2θ of 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, and 10.3±0.2°.

10. The acid salt according to claim 9, characterized in that, The X-ray powder diffraction pattern of the phosphate crystal form A may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, 21.4±0.2°, 17.0±0.2°, and 24.4±0.2°. The X-ray powder diffraction pattern of the hydrochloride crystal form A may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, 29.1±0.2°, 14.3±0.2°, and 13.6±0.2°. The X-ray powder diffraction pattern of the methanesulfonate crystal form A may optionally include 2-3, 4-5, or 6-7 diffraction peaks located at any of the following 2θ values: 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, 10.3±0.2°, 18.3±0.2°, and 24.6±0.2°.

11. The acid salt according to claim 10, characterized in that, The X-ray powder diffraction pattern of the phosphate crystal form A may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, 21.4±0.2°, 17.0±0.2°, and 24.4±0.2°. The X-ray powder diffraction pattern of the hydrochloride crystal form A may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, 29.1±0.2°, 14.3±0.2°, and 13.6±0.2°. The X-ray powder diffraction pattern of the methanesulfonate crystal form A may optionally include diffraction peaks at any 2, 3, 4, 5, 6, or 7 locations within the range of 2θ values ​​of 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, 10.3±0.2°, 18.3±0.2°, and 24.6±0.2°.

12. The acid salt according to claim 9, characterized in that, The X-ray powder diffraction pattern of the phosphate crystal form A includes 10 diffraction peaks selected from 2θ values ​​of 13.8±0.2°, 23.4±0.2°, 21.0±0.2°, 25.4±0.2°, 20.3±0.2°, 10.1±0.2°, 9.5±0.2°, 21.4±0.2°, 17.0±0.2°, 24.4±0.2°, 22.9±0.2°, 20.5±0.2°, 22.6±0.2°, 17.5±0.2°, and 13.5±0.2°. The X-ray powder diffraction pattern of the hydrochloride crystal form A includes 10 diffraction peaks located at 2θ values ​​of 22.7±0.2°, 19.4±0.2°, 25.3±0.2°, 22.3±0.2°, 25.7±0.2°, 8.2±0.2°, 17.9±0.2°, 29.1±0.2°, 14.3±0.2°, 13.6±0.2°, 16.3±0.2°, 30.8±0.2°, 23.8±0.2°, 20.7±0.2°, and 27.5±0.2°. The X-ray powder diffraction pattern of the methanesulfonate crystal form A contains diffraction peaks at any of the following 10 locations with 2θ values ​​of 20.9±0.2°, 6.2±0.2°, 13.4±0.2°, 22.4±0.2°, 25.6±0.2°, 14.8±0.2°, 18.9±0.2°, 10.3±0.2°, 18.3±0.2°, 24.6±0.2°, 8.2±0.2°, 12.8±0.2°, 31.9±0.2°, 9.5±0.2°, and 29.6±0.2°.

13. The acid salt according to claim 9, characterized in that, The X-ray powder diffraction pattern of the phosphate crystal form A is shown in Figure 22, and the DSC pattern is shown in Figure 23. The X-ray powder diffraction pattern of the hydrochloride crystal form A is shown in Figure 24, and the DSC pattern is shown in Figure 25. The X-ray powder diffraction pattern of the methanesulfonate crystal form A is shown in Figure 26, and the DSC pattern is shown in Figure 27.

14. The acid salt according to claim 13, characterized in that, The positions of the top ten diffraction peaks with the highest relative intensities in the X-ray powder diffraction patterns of phosphate crystal form A, hydrochloride crystal form A, and methanesulfonate crystal form A have a 2θ error of ±0.2° compared with the diffraction peaks at the corresponding positions in Figures 22, 24, and 26, respectively.

15. A method for preparing the acid salt according to any one of claims 8-14, specifically comprising the following steps: 1) Weigh an appropriate amount of the free base of compound I, and optionally dissolve it in a good or bad solvent; 2) Weigh an appropriate amount of the counterionic acid and dissolve it in an organic solvent; the amount of the counterionic acid is 1.2 equivalents. 3) Combine the two solutions and stir to precipitate. 4) Rapid centrifugation or static drying to obtain the target product; in: The benign solvent is selected from methanol or ethanol; The unsuitable solvent is selected from ethyl acetate, acetone, acetonitrile, or 2-methyltetrahydrofuran; The organic solvent is selected from methanol, ethanol, or acetonitrile; the above-mentioned benign solvents and organic solutions must be miscible when used. The aforementioned counterionic acid is selected from phosphoric acid, hydrochloric acid, or methanesulfonic acid.

16. A pharmaceutical composition comprising a therapeutically effective amount of the crystal form of any one of claims 1-6, or the acid salt or crystal form of any one of claims 8-14, and one or more pharmaceutically acceptable carriers or excipients.

17. The use of the crystal form of the compound according to any one of claims 1-6, or the acid salt or crystal form of the compound according to any one of claims 8-14, and the use of the pharmaceutical composition according to claim 16 in the preparation of MEK inhibitors, EGFR inhibitors and EGFR monoclonal antibodies and their combination with related drugs.

18. The use of the crystal form of the compound according to any one of claims 1-6, or the acid salt or crystal form of the compound according to any one of claims 8-14, and the use of the pharmaceutical composition according to claim 16 in the preparation of a medicament for treating cancer-related diseases; wherein the cancer is selected from breast cancer, cervical cancer, colon cancer, lung cancer, gastric cancer, rectal cancer, pancreatic cancer, brain cancer, liver cancer, solid tumor, glioma, glioblastoma, leukemia, lymphoma, or myeloma.

19. The application according to claim 18, characterized in that, The cancer in question was selected from non-small cell lung cancer.

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