Salt of 3,4-dihydroisoquinoline compound and application thereof

Abstract

Salts of the compound shown in formula (A), their solid forms and crystal forms, pharmaceutical compositions comprising them, their uses, and methods of preparation are provided. Compared to the compound of formula (A), the salts of the compound shown in formula (A) have higher solubility in water and biological solvents, are easier to prepare in solid form, are easier to transfer and weigh, and have good application prospects.

Description

[0001] This invention claims priority to Chinese Patent Application No. 202111494667.8, filed on December 8, 2021. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical technology and relates to salts of 3,4-dihydroisoquinoline compounds, their crystalline forms and specific crystal forms, pharmaceutical compositions containing them, and their applications in the pharmaceutical field. Background Technology

[0003] Protein arginine methyltransferases (PRMTs) are a class of S-adenosylmethionine (SAM or AdoMet)-dependent methyltransferases that catalyze the methylation of proteins, specifically transferring methyl groups from AdoMet to the guanidino nitrogen atom at the terminus of arginine residues in histones or other proteins. PRMTs play important roles in protein methylation, such as participating in alternative splicing, post-transcriptional regulation, RNA processing, cell proliferation, cell differentiation, apoptosis, and tumorigenesis. Based on their catalytic arginine methylation mechanisms, the PRMT family can be divided into three types: PRMT1-4, PRMT6, and PRMT8 belong to type I, catalyzing monomethyl and asymmetric dimethyl methylation; PRMT5 and PRMT9 belong to type II, catalyzing symmetric dimethyl methylation; and PRMT7 belongs to type III, capable of monomethyl catalysis.

[0004] PRMT5 was first isolated by Pollack et al. from a protein complex bound to Jak2 (Janus tyrosine kinase 2) in yeast two-hybrid studies, and is therefore also known as JBP1 (jak-binding protein 1). PRMT5 not only regulates gene transcription and protein modification, but also plays a role in regulating cell proliferation, differentiation, and apoptosis during tumor cell growth, making it a highly promising target for cancer therapy. Currently, the development of PRMT5 inhibitors is in its early stages. The most advanced is GSK3326595, released by GSK, which is in Phase I / II clinical trials. JNJ-64619178, first released by Janssen, PF-06939999 by Pfizer, and PRT-543 by Prelude Therapeutics are all in Phase I clinical trials. Currently, the structural formulas of PF-06939999 and PRT-543 have not been published. The structural formulas of GSK3326595 and JNJ-64619178 are as follows:

[0005]

[0006] Given that no PRMT5 inhibitors have yet been approved for marketing, designing and synthesizing novel PRMT5 inhibitors that combine good efficacy and good dosing performance will have significant clinical application value. Summary of the Invention

[0007] In a first aspect, the present invention provides a salt of the compound shown in formula (A),

[0008]

[0009] The salt is an inorganic acid addition salt or an organic acid addition salt.

[0010] According to some embodiments of the present invention, the inorganic acid addition salt is a sulfate or a phosphate.

[0011] According to some embodiments of the present invention, the organic acid addition salt is selected from malate, oxalate, succinate, tartrate, adipate, citrate, gluconate, maleate, fumarate, lactate, and gentianate; preferably malate, oxalate, succinate, L-tartrate, L-lactate, adipate, citrate, or gluconate; more preferably malate, oxalate, succinate, L-tartrate, adipate, citrate, or gluconate; further preferably malate, oxalate, citrate, or gluconate; even more preferably malate or oxalate; and even more preferably L-malate or oxalate.

[0012] According to some embodiments of the present invention, in the salt of the compound represented by formula (A), the chemical ratio of the compound represented by formula (A) to the organic or inorganic acid molecule is 1:0.5 to 2; preferably 1:1 to 1.5; more preferably 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4 or 1:1.5; more preferably 1:1, 1:1.3, 1:1.4 or 1:1.5; more preferably 1:1 or 1:1.5; more preferably 1:1.

[0013] According to some embodiments of the present invention, the salt of the compound shown in formula (A) is the compound shown in formula (A-1).

[0014]

[0015] Wherein, X is an inorganic acid or an organic acid, and n is selected from 0.5-2; preferably 1-1.5; further preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; further preferably 1, 1.3, 1.4 or 1.5; further preferably 1:1 or 1:1.5; even more preferably 1.

[0016] According to some embodiments of the present invention, X is an inorganic acid selected from sulfuric acid and phosphoric acid.

[0017] According to some embodiments of the present invention, X is an organic acid selected from malic acid, oxalic acid, succinic acid, tartaric acid, adipic acid, citric acid, gluconic acid, maleic acid, fumaric acid, lactic acid, and gentianic acid; preferably malic acid, oxalic acid, succinic acid, L-tartaric acid, L-lactic acid, adipic acid, citric acid, or gluconic acid; more preferably malic acid, oxalic acid, succinic acid, L-tartaric acid, adipic acid, citric acid, or gluconic acid; even more preferably malic acid or oxalic acid; even more preferably L-malic acid or oxalic acid.

[0018] According to some embodiments of the present invention, the salt of the compound shown in formula (A) or the compound shown in formula (A-1) is in solid form.

[0019] According to some embodiments of the present invention, the salt of the compound of formula (A) or the compound of formula (A-1) in solid form is in crystalline form.

[0020] According to some embodiments of the present invention, the compound represented by formula (A-1) is a compound represented by formula (B):

[0021]

[0022] Wherein, n is selected from 0.5-2; preferably 1-1.5; further preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; preferably 1 or 1.5.

[0023] According to some embodiments of the present invention, the compound represented by formula (B) is in solid form.

[0024] According to some embodiments of the present invention, the compound of formula (B) in solid form, when subjected to KBr pelleting, has an infrared spectrum comprising characteristic peaks at the following positions (±4 cm⁻¹). -1 ): 3301, 2940, 1611, 1528, 1456, 1368, 1045.

[0025] According to some embodiments of the present invention, the compound of formula (B) in solid form is in crystalline form.

[0026] According to some embodiments of the present invention, the compound represented by formula (B) is a compound represented by formula (B-1):

[0027]

[0028] According to some embodiments of the present invention, the compound represented by formula (B-1) is in solid form.

[0029] According to some embodiments of the present invention, the compound of formula (B-1) in solid form is in crystalline form.

[0030] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form I, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.2, 6.5, 13.3, 19.1.

[0031] According to some embodiments of the present invention, the crystal form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.2, 6.5, 13.3, 19.1, 20.1, 22.0.

[0032] According to some embodiments of the present invention, the crystal form I, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.2, 6.5, 13.3, 18.3, 19.1, 20.1, 22.0.

[0033] According to some embodiments of the present invention, crystal form I, using Cu-Kα radiation, has substantially the following properties: Figure 1 The X-ray powder diffraction pattern shown is shown.

[0034] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form I has an endothermic peak at 113±5℃.

[0035] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form I has endothermic peaks at 102.15±5℃ and 113±5℃.

[0036] According to some embodiments of the present invention, the crystal form I has a thermogravimetric analysis curve showing a weight loss of 8.6522% ± 0.2% between room temperature and 110 ± 5°C.

[0037] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form II, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.9, 6.6, 13.2, 18.7, 19.8.

[0038] According to some embodiments of the present invention, the crystal form II, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.9, 6.6, 13.2, 18.7, 19.8, 22.1, 26.5.

[0039] According to some embodiments of the present invention, the crystal form II, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.9, 6.6, 13.2, 18.7, 19.8, 22.1, 23.3, 26.5.

[0040] According to some embodiments of the present invention, the crystal form II, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.9, 6.6, 13.2, 18.7, 19.8, 20.3, 22.1, 23.3, 26.5.

[0041] According to some embodiments of the present invention, the crystal form II, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.9, 6.6, 12.3, 13.2, 18.7, 19.8, 20.3, 22.1, 23.3, 24.0, 26.5.

[0042] According to some embodiments of the present invention, the crystal form II, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.9, 6.6, 12.3, 13.2, 18.7, 19.8, 20.3, 22.1, 23.3, 24.0, 26.5, 28.8.

[0043] According to some embodiments of the present invention, crystal form II, using Cu-Kα radiation, has substantially the following properties: Figure 2 The X-ray powder diffraction pattern shown is shown.

[0044] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form II has an endothermic peak at 103.58±5℃.

[0045] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form II has endothermic peaks at 89.8±5℃ and 103.58±5℃.

[0046] According to some embodiments of the present invention, the crystal form II has a thermogravimetric analysis curve showing a weight loss of 2.6358% ± 0.2% between room temperature and 80 ± 5°C.

[0047] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form III, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.3, 6.9, 20.3.

[0048] According to some embodiments of the present invention, the crystal form III, using Cu-Kα radiation, has X-ray powder diffraction patterns with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.3, 6.9, 8.8, 20.3, 21.2.

[0049] According to some embodiments of the present invention, the crystal form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.3, 6.9, 8.8, 12.8, 13.3, 20.3, 21.2.

[0050] According to some embodiments of the present invention, the crystal form III, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.3, 6.9, 8.8, 12.8, 13.3, 14.0, 15.9, 20.3, 21.2.

[0051] According to some embodiments of the invention, crystal form III, using Cu-Kα radiation, has essentially the following properties: Figure 3 The X-ray powder diffraction pattern shown is shown.

[0052] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form III has an endothermic peak at 117.93±5℃.

[0053] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form III has endothermic peaks at 70.82±5℃ and 117.93±5℃.

[0054] According to some embodiments of the present invention, the crystal form III has a thermogravimetric analysis curve showing a weight loss of 3.0001% ± 0.2% between room temperature and 75 ± 5°C.

[0055] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form IV, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.4, 7.6, 8.9, 13.9, 20.6.

[0056] According to some embodiments of the present invention, the crystal form IV, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.4, 7.6, 8.9, 12.1, 13.9, 15.2, 20.6.

[0057] According to some embodiments of the present invention, the crystal form IV, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.4, 7.6, 8.9, 12.1, 13.9, 15.2, 17.8, 18.5, 20.6.

[0058] According to some embodiments of the present invention, the crystal form IV, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.4, 7.6, 8.9, 12.1, 13.9, 15.2, 17.1, 17.8, 18.5, 20.6.

[0059] According to some embodiments of the present invention, the crystal form IV, using Cu-Kα radiation, has substantially the following properties: Figure 4 The X-ray powder diffraction pattern shown is shown.

[0060] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form IV has an endothermic peak at 113.27±5℃.

[0061] According to some embodiments of the present invention, the differential scanning calorimetry curve of crystal form IV has endothermic peaks at 77.91±5℃ and 113.27±5℃.

[0062] According to some embodiments of the present invention, the crystal form IV has a thermogravimetric analysis curve showing a weight loss of 2.6271% ± 0.2% between room temperature and 60 ± 5°C.

[0063] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form V, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 5.0, 13.6, 18.6, 19.6, 20.2.

[0064] According to some embodiments of the present invention, the crystal form V, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.0, 6.8, 13.1, 13.6, 18.6, 19.6, 20.2.

[0065] According to some embodiments of the present invention, the crystal form V, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.0, 6.8, 13.1, 13.6, 16.7, 18.6, 19.6, 20.2, 24.3.

[0066] According to some embodiments of the present invention, the crystal form V, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.0, 6.8, 13.1, 13.6, 16.7, 18.6, 19.6, 20.2, 23.2, 24.3, 25.0.

[0067] According to some embodiments of the present invention, the crystal form V, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 5.0, 6.8, 13.1, 13.6, 16.7, 18.6, 19.6, 20.2, 23.2, 24.3, 25.0, 28.6.

[0068] According to some embodiments of the present invention, the crystal form V, using Cu-Kα radiation, has substantially the following properties: Figure 5 The X-ray powder diffraction pattern shown is shown.

[0069] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form V has an endothermic peak at 104.69±5℃.

[0070] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form V has endothermic peaks at 68.38±5℃ and 104.69±5℃.

[0071] According to some embodiments of the present invention, the crystal form V has a thermogravimetric analysis curve showing a weight loss of 3.6041% ± 0.2% between room temperature and 75 ± 5°C.

[0072] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form VI, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.5, 7.0, 9.0, 12.9, 20.2, 21.6.

[0073] According to some embodiments of the present invention, the crystal form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.5, 7.0, 9.0, 12.9, 13.3, 15.8, 20.2, 21.6.

[0074] According to some embodiments of the present invention, the crystal form VI, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.5, 7.0, 9.0, 12.9, 13.3, 13.9, 15.8, 20.2, 21.6.

[0075] According to some embodiments of the present invention, the crystal form VI, when irradiated with Cu-Kα, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.5, 7.0, 9.0, 12.9, 13.3, 13.9, 15.8, 16.9, 20.2, 21.6, 25.4.

[0076] According to some embodiments of the present invention, crystal form VI, using Cu-Kα radiation, has substantially the following properties: Figure 6 The X-ray powder diffraction pattern shown is shown.

[0077] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form VI has an endothermic peak at 113.29±5℃.

[0078] According to some embodiments of the present invention, the crystal form VI has a thermogravimetric analysis curve showing a weight loss of 0.34% ± 0.2% between room temperature and 120 ± 5°C.

[0079] According to some embodiments of the present invention, the crystalline form of the compound of formula (B), which is crystal form VII, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.3, 6.9, 13.2, 19.1, 20.0.

[0080] According to some embodiments of the present invention, the crystal form VII, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.3, 6.9, 8.9, 13.2, 19.1, 20.0, 21.7.

[0081] According to some embodiments of the present invention, the crystal form VII, using Cu-Kα radiation, has an X-ray powder diffraction pattern with characteristic diffraction peaks (±0.2°) at the following 2θ angles: 4.3, 6.9, 8.9, 13.2, 15.1, 19.1, 20.0, 21.1, 21.7.

[0082] According to some embodiments of the invention, crystal form VII, using Cu-Kα radiation, has substantially the following properties: Figure 7 The X-ray powder diffraction pattern shown is shown.

[0083] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form VII has an endothermic peak at 197.3±5℃.

[0084] According to some embodiments of the present invention, the differential scanning calorimetry curve of the crystal form VII has endothermic peaks at 92.4±5℃ and 197.3±5℃.

[0085] According to some embodiments of the present invention, the crystal form VII has a thermogravimetric analysis curve showing a weight loss of 7.88% ± 0.2% between room temperature and 110 ± 5 °C.

[0086] According to some embodiments of the present invention, the compound represented by formula (A-1) is a compound represented by formula (C):

[0087]

[0088] Wherein, n is selected from 0.5-2; preferably 1-1.5; further preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; even more preferably 1 or 1.5.

[0089] According to some embodiments of the present invention, the compound represented by formula (C) is in solid form.

[0090] According to some embodiments of the present invention, the compound of formula (C) in solid form, when subjected to KBr pelleting, has an infrared spectrum comprising characteristic peaks at the following positions (±4 cm⁻¹). -1 ): 3320, 2937, 1615, 1526, 1456, 1368, 1047.

[0091] According to some embodiments of the present invention, the compound of formula (C) in solid form is in crystalline form.

[0092] According to some embodiments of the present invention, the compound represented by formula (C) is a compound represented by formula (C-1) or formula (C-2):

[0093]

[0094] According to some embodiments of the present invention, the compound represented by formula (C-1) or the compound represented by formula (C-2) is in solid form.

[0095] According to some embodiments of the present invention, the compound of formula (C-1) in solid form and the compound of formula (C-2) in solid form are in crystalline form.

[0096] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form I, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.7, 7.1, 10.7, 17.4, 21.2.

[0097] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form I, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.7, 7.1, 10.7, 16.1, 17.4, 20.1, 21.2.

[0098] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form I, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.7, 7.1, 10.7, 16.1, 17.4, 19.6, 20.1, 21.2, 23.2.

[0099] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form I, exhibits substantially the following properties when subjected to Cu-Kα irradiation: Figure 8 The X-ray powder diffraction pattern shown is shown.

[0100] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form II, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 13.0, 21.6.

[0101] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form II, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 13.0, 21.6, 24.5, 25.0.

[0102] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form II, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 4.8, 13.0, 18.4, 19.4, 21.6, 24.5, 25.0.

[0103] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form II, exhibits substantially the following properties when subjected to Cu-Kα irradiation: Figure 9 The X-ray powder diffraction pattern shown is shown.

[0104] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C) is crystal form II, and its differential scanning calorimetry curve has an endothermic peak at 162.6 ± 5 °C.

[0105] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form II, has a thermogravimetric analysis curve showing a weight loss of 3.2% ± 0.2% between room temperature and 160 ± 5 °C.

[0106] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form III, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 18.7, 19.4.

[0107] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form III, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 4.8, 14.0, 16.7, 18.7, 19.4, 23.3.

[0108] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form III, exhibits substantially the following properties when subjected to Cu-Kα irradiation: Figure 10 The X-ray powder diffraction pattern shown is shown.

[0109] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C) is crystal form III, and its differential scanning calorimetry curve has an endothermic peak at 204.9 ± 5 °C.

[0110] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form III, has endothermic peaks at 139.9±5℃ and 204.9±5℃ in its differential scanning calorimetry curve.

[0111] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form III, has a thermogravimetric analysis curve showing a weight loss of 6.2% ± 0.2% between room temperature and 150 ± 5 °C.

[0112] According to some embodiments of the present invention, the crystalline form of the compound of formula (C), which is crystal form IV, has a characteristic diffraction peak (±0.2°) at the following 2θ angles when subjected to Cu-Kα radiation: 4.6, 13.7, 19.5, 20.0, 22.9.

[0113] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form IV, exhibits substantially the following properties when subjected to Cu-Kα irradiation: Figure 11 The X-ray powder diffraction pattern shown is shown.

[0114] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C) is crystal form IV, and its differential scanning calorimetry curve has an endothermic peak at 137.4 ± 5 ​​°C.

[0115] According to some embodiments of the present invention, the crystalline form of the compound represented by formula (C), which is crystal form IV, has a thermogravimetric analysis curve showing a weight loss of 4.26 ± 0.2% between room temperature and 150 ± 5 °C.

[0116] According to some embodiments of the present invention, the compound of formula (C) in crystalline form V, when subjected to Cu-Kα irradiation, yields a single crystal belonging to the triclinic crystal system, space group P1, with unit cell parameters as follows: { α=99.1280(10)°, β=90.1780(10)°, γ=95.1340(10)°, }

[0117] In a second aspect, the present invention provides a crystalline composition comprising one or more of the following crystal forms: crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V, crystal form VI, and crystal form VII of the compound represented by formula (B).

[0118] According to some embodiments of the present invention, in the crystalline composition, the crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V, crystal form VI or crystal form VII of the compound represented by formula (B) accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more of the weight of the crystalline composition.

[0119] Thirdly, the present invention provides a crystalline composition comprising one or more of the following crystal forms: crystal form I, crystal form II, crystal form III, crystal form IV, and crystal form V of the compound represented by formula (C).

[0120] According to some embodiments of the present invention, in the crystalline composition, the crystal form I, crystal form II, crystal form III, crystal form IV or crystal form V of the compound represented by formula (C) accounts for more than 50%, more than 60%, more than 70%, more than 80%, more than 90% or more or 95% of the weight of the crystalline composition.

[0121] Fourthly, the present invention provides a pharmaceutical composition comprising a salt of the compound shown in formula (A), the compound shown in formula (A-1), the compound shown in solid form (A-1), the compound shown in crystalline form (A-1), the compound shown in formula (B), the compound shown in solid form (B), the compound shown in crystalline form (B), the compound shown in formula (B-1), the compound shown in solid form (B-1), the compound shown in crystalline form (B-1), crystal form I of the compound shown in formula (B), crystal form II of the compound shown in formula (B), crystal form III of the compound shown in formula (B), crystal form IV of the compound shown in formula (B), crystal form V of the compound shown in formula (B), and the compound shown in formula (B). Crystal form VI of the compound shown, crystal form VII of the compound shown in formula (B), the compound shown in formula (C), the compound shown in solid form of formula (C), the compound shown in crystalline form of formula (C), the compound shown in formula (C-1), the compound shown in formula (C-2), the compound shown in solid form of formula (C-1) or the compound shown in formula (C-2), the compound shown in crystalline form of formula (C-1) or the compound shown in formula (C-2), crystal form I of the compound shown in formula (C), crystal form II of the compound shown in formula (C), crystal form III of the compound shown in formula (C), crystal form IV of the compound shown in formula (C), crystal form V of the compound shown in formula (C), or the crystalline composition described in the second or third aspect above.

[0122] According to some embodiments of the present invention, the above-described pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0123] Fifthly, the present invention provides salts of the compounds represented by formula (A) mentioned in the foregoing aspects, compounds represented by formula (A-1), compounds represented by formula (A-1) in solid form, compounds represented by formula (A-1) in crystalline form, compounds represented by formula (B), compounds represented by formula (B) in solid form, compounds represented by formula (B-1), compounds represented by formula (B-1) in solid form, compounds represented by formula (B-1) in crystalline form, crystal form I of the compound represented by formula (B), crystal form II of the compound represented by formula (B), crystal form III of the compound represented by formula (B), crystal form IV of the compound represented by formula (B), crystal form V of the compound represented by formula (B), crystal form VI of the compound represented by formula (B), and other crystal forms of the compound represented by formula (B). The following are examples of the use of the following compounds: crystal form VII, the compound of formula (C), the compound of formula (C) in solid form, the compound of formula (C) in crystalline form, the compound of formula (C-1), the compound of formula (C-2), the compound of formula (C-1) or the compound of formula (C-2) in solid form, the compound of formula (C-1) or the compound of formula (C-2) in crystalline form, crystal form I, crystal form II, crystal form III, crystal form IV, crystal form V of the compound of formula (C), the crystalline composition described in the second or third aspect above, or the pharmaceutical composition described in the fourth aspect above, as a medicament or in the preparation of a medicament.

[0124] According to some embodiments of the present invention, the drug is used for the prevention and / or treatment of proliferative diseases; preferably, the proliferative disease is a tumor or cancer; more preferably, the tumor or cancer is a hematologic malignancy or a solid tumor; even more preferably, it is a malignant hematologic malignancy or an advanced solid tumor; even more preferably, it is a relapsed or refractory hematologic malignancy or an advanced malignant solid tumor.

[0125] According to some embodiments of the present invention, the drug is used to prevent and / or treat diseases mediated at least in part by PRMT5.

[0126] In a sixth aspect, the present invention provides salts of the compounds represented by formula (A) mentioned in the foregoing aspects, compounds represented by formula (A-1), compounds represented by formula (A-1) in solid form, compounds represented by formula (A-1) in crystalline form, compounds represented by formula (B), compounds represented by formula (B) in solid form, compounds represented by formula (B) in crystalline form, compounds represented by formula (B-1), compounds represented by formula (B-1) in solid form, compounds represented by formula (B-1) in crystalline form, crystal form I of the compound represented by formula (B), crystal form II of the compound represented by formula (B), crystal form III of the compound represented by formula (B), crystal form IV of the compound represented by formula (B), crystal form V of the compound represented by formula (B), crystal form VI of the compound represented by formula (B), and crystal form VI of the compound represented by formula (B). I. The compound of formula (C), the compound of formula (C) in solid form, the compound of formula (C) in crystalline form, the compound of formula (C-1), the compound of formula (C-2), the compound of formula (C-1) or the compound of formula (C-2) in solid form, the compound of formula (C-1) or the compound of formula (C-2) in crystalline form, crystal form I of the compound of formula (C), crystal form II of the compound of formula (C), crystal form III of the compound of formula (C), crystal form IV of the compound of formula (C), crystal form V of the compound of formula (C), the crystalline composition described in the second or third aspect above, or the pharmaceutical composition described in the fourth aspect above, for the prevention and / or treatment of diseases or cell proliferative disorders mediated at least in part by PRMT5.

[0127] In a seventh aspect, the present invention provides a method for preventing and / or treating at least partially PRMT5-mediated diseases or proliferative disorders, comprising: administering to an individual in need a therapeutically effective amount of a salt of the compound of formula (A), the compound of formula (A-1), the compound of formula (A-1) in solid form, the compound of formula (A-1) in crystalline form, the compound of formula (B), the compound of formula (B) in solid form, the compound of formula (B) in crystalline form, the compound of formula (B-1), the compound of formula (B-1) in solid form, the compound of formula (B-1) in crystalline form, crystal form I of the compound of formula (B), crystal form II of the compound of formula (B), crystal form III of the compound of formula (B), and crystal form of the compound of formula (B). IV. Crystal form V of the compound shown in formula (B), VI of the compound shown in formula (B), VII of the compound shown in formula (B), the compound shown in formula (C), the compound shown in formula (C) in solid form, the compound shown in formula (C) in crystalline form, the compound shown in formula (C-1), the compound shown in formula (C-2), the compound shown in formula (C-1) or the compound shown in formula (C-2) in solid form, the compound shown in formula (C-1) or the compound shown in formula (C-2) in crystalline form, the compound shown in formula (C) in crystal form I, the compound shown in formula (C) in crystal form II, the compound shown in formula (C) in crystal form III, the compound shown in formula (C) in crystal form IV, the compound shown in formula (C) in crystal form V, the crystalline composition described in the second or third aspect above, or the pharmaceutical composition described in the fourth aspect above.

[0128] According to some embodiments of the present invention, the diseases at least partially mediated by PRMT5 described in the fifth, sixth or seventh aspects above are proliferative diseases.

[0129] According to some embodiments of the present invention, the cell proliferative disease described in the fifth, sixth or seventh aspects above is a tumor or cancer; preferably, the tumor or cancer is a hematologic malignancy or a solid tumor; more preferably, it is a malignant hematologic malignancy or an advanced solid tumor; and even more preferably, it is a relapsed or refractory hematologic malignancy or an advanced malignant solid tumor.

[0130] According to some embodiments of the present invention, the tumors or cancers described in the fifth, sixth, or seventh aspects above are selected from lung cancer, bone cancer, gastric cancer, pancreatic cancer, adenoid cystic carcinoma, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma, and chronic and acute leukemia; preferably, the acute leukemia is acute myeloid leukemia (AML).

[0131] Eighthly, the present invention provides a method for preparing the compound of formula (A-1), comprising reacting the compound of formula (A) with an acid in a suitable solvent to obtain the compound of formula (A-1):

[0132]

[0133] Wherein, X is an acid; preferably an inorganic acid or an organic acid; n is selected from 0.5-2.

[0134] According to some embodiments of the present invention, n is selected from 1 to 1.5; preferably 1, 1.1, 1.2, 1.3, 1.4 or 1.5; more preferably 1, 1.3, 1.4 or 1.5; and even more preferably 1.

[0135] According to some embodiments of the present invention, X is an inorganic acid selected from sulfuric acid and phosphoric acid.

[0136] According to some embodiments of the present invention, X is an organic acid selected from malic acid, oxalic acid, succinic acid, tartaric acid, adipic acid, citric acid, gluconic acid, maleic acid, fumaric acid, lactic acid, and gentianic acid; preferably malic acid, oxalic acid, succinic acid, L-tartaric acid, L-lactic acid, adipic acid, citric acid, or gluconic acid; more preferably malic acid, oxalic acid, succinic acid, L-tartaric acid, adipic acid, citric acid, or gluconic acid; even more preferably malic acid or oxalic acid; even more preferably L-malic acid or oxalic acid.

[0137] According to the preparation method of the present invention, the molar ratio of the compound represented by formula (A) to the acid is 1-2:0.5-2, preferably 1:1-2, and more preferably 1:1.1-2.

[0138] According to the preparation method of the present invention, the reaction temperature is 0-90℃, preferably 5-80℃, more preferably 20-60℃, and even more preferably room temperature-50℃.

[0139] According to the preparation method of the present invention, the reaction solvent is selected from one or a combination of two of alcohols, esters, nitriles, ketones, water, alkane solvents, ether solvents, or heterocyclic alkane solvents; preferably, it is selected from one or a combination of two of ROH, RCOOR1, RCN, RCOOR1, water, ROR1, RH, or heterocyclic alkane solvents, wherein R and R1 are each independently selected from C 1-6 Straight-chain or branched alkyl groups; preferably, R and R1 are each independently selected from C1. 1-4The reaction solvent is a straight-chain or branched alkyl group; preferably, the reaction solvent is selected from one or a combination of two of isopropanol, methanol, ethanol, ethyl acetate, acetone, butanone, acetonitrile, water, tetrahydrofuran, n-heptane, and 2-methyltetrahydrofuran; when it is a mixed solvent composed of two solvents, the volume ratio of the two solvents is 1-20:20-1, preferably 1-19:19-1, and more preferably 1-10:10-1.

[0140] According to the preparation method of the present invention, after the reaction is completed, optionally, the temperature is lowered to -15 to 15°C, and the mixture is allowed to stand for crystallization for 0.5 h to 5 days. The solid is then separated, dried, and the compound shown in formula (A-1) is obtained. Preferably, the crystallization temperature is 5°C, and the crystallization time is 1 h to 3 days.

[0141] According to the preparation method of the present invention, the separation step includes using suitable methods such as suction filtration, vacuum filtration, filtration, and centrifugation to separate the compound represented by the obtained formula (A-1) from the crystallization liquid.

[0142] According to the preparation method of the present invention, the drying method can employ any suitable known method, preferably room temperature drying, room temperature vacuum drying, or drying at 50°C. Specific drying conditions include, for example, a drying time preferably of 1 hour to 5 days, more preferably 3 hours to 3 days, and even more preferably 3 hours to 1 day. Regardless of the drying method used, it is preferable that the solvent residue in the resulting product meets the quality standards.

[0143] Definitions and Explanations

[0144] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense.

[0145] The compound represented by formula (A) mentioned in this invention may exist in specific stereoisomer forms, including cis and trans isomers, (R)- and (S)-enantiomers, and racemic mixtures thereof, with trans isomers being preferred.

[0146] The terms "salt of the compound shown in formula (A) or the compound shown in formula (A-1) in solid form", "compound shown in formula (B) in solid form", or "compound shown in formula (C) in solid form" mentioned in this invention refer to compounds shown in formula (A-1), compounds shown in formula (B), compounds shown in formula (C) in solid form, etc., including crystalline and amorphous forms of compounds shown in formula (A-1), compounds shown in formula (B), compounds shown in formula (C), or compounds shown in formula (C-2).

[0147] The terms "salt of the compound shown in formula (A) in crystalline form or compound shown in formula (A-1)", "compound shown in formula (B) in crystalline form" or "compound shown in formula (C) in crystalline form" mentioned in this invention refer to compounds shown in formula (A-1), formula (B), and formula (C) in crystalline form, including anhydrous and solvent-free forms, hydrate forms, solvate forms, and eutectic forms of compounds shown in formula (A-1), formula (B), and formula (C).

[0148] The term "solvent" or "solvent compound" refers to an association formed by stoichiometric or non-stoichiometric solvent molecules with a salt of the compound shown in formula (A), the compound shown in formula (A-1), the compound shown in formula (B), the compound shown in formula (C), etc. of the present invention, including associations containing both water molecules and one or more other solvent molecules, and associations containing only one or more other solvent molecules.

[0149] The term "hydrate" refers to an assemblage formed by water molecules in stoichiometric or non-stoichiometric proportions with a salt of the compound shown in formula (A), the compound shown in formula (A-1), the compound shown in formula (B), the compound shown in formula (C), etc.

[0150] The term "anhydrous and solvent-free form" refers to a form that does not contain water or solvent molecules, or where water or solvent molecules coexist with the salt of the compound shown in formula (A), the compound shown in formula (A-1), the compound shown in formula (B), the compound shown in formula (C), etc., in a manner not bound by intermolecular forces, such as through adsorption.

[0151] The term "crystalline composition" refers to a solid form comprising one or more specific crystal forms of the compounds mentioned in this invention (salts of the compound shown in formula (A), compounds shown in formula (A-1), compounds shown in formula (B), compounds shown in formula (C), etc.). For example, in one embodiment of this invention, it comprises one, two, or more of crystal forms I, II, III, IV, V, VI, and VII of the compound shown in formula (B). Furthermore, in addition to the crystal forms of this invention, the crystalline composition may optionally contain other crystalline forms, other crystal forms, or other amorphous forms of compounds (compounds shown in formula (A-1), (B), (B-1), (C), (C-1), or (C-2)), or impurities other than these substances. Those skilled in the art will understand that the sum of the contents of all components in the crystalline composition should be 100%.

[0152] The term "room temperature" refers to room temperature in the conventional sense of the art, generally 10 to 30°C, preferably 25°C ± 5°C.

[0153] In the context of this invention, the 2θ values ​​in X-ray powder diffraction patterns are all in degrees (°).

[0154] In X-ray powder diffraction (XRD) patterns, the term "substantially" or "substantially as shown" refers to a substantially pure crystalline form whose powder XRD pattern shows at least 50%, or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99% of the peaks. Furthermore, as the content of a crystalline form in a product gradually decreases, some diffraction peaks attributable to that crystalline form in the XRD pattern may decrease due to factors such as instrument detection sensitivity. In addition, for any given crystalline form, the peak positions may have slight errors, which is well known in the field of crystallography. For example, due to temperature variations during sample analysis, sample movement, or instrument calibration, the peak positions can shift, and the determination error of the 2θ value is typically about ±0.2°. Therefore, this error should be taken into account when determining each crystal structure, and the terms "substantially" or "substantially as shown" are also intended to cover such differences in diffraction peak positions.

[0155] In DSC or TGA spectra, the terms "substantially" or "substantially as shown" refer to the fact that, for the same compound and the same crystal form, the error in thermal transition onset temperature, endothermic peak peak temperature, exothermic peak peak temperature, melting point, weight loss onset temperature, or weight loss endpoint temperature, etc., is typically within about 5°C, and usually within about 3°C, in consecutive analyses. When describing a compound as having a given thermal transition onset temperature, endothermic peak peak temperature, exothermic peak peak temperature, melting point, weight loss onset temperature, or weight loss endpoint temperature, it refers to that temperature ±5°C.

[0156] As used in this article, “cell proliferation disorder” refers to a condition in which the cell population grows at a rate lower or higher than the expected rate under given physiological conditions.

[0157] The term "tumor" includes benign tumors, malignant tumors, and borderline tumors, with malignant tumors collectively referred to as cancer.

[0158] As used herein, the term "prevention" means that, when used for a disease or condition (e.g., cancer), the compound or drug reduces the frequency of symptoms of the medical condition or delays its onset in a subject compared to a subject who has not been given the compound or drug (e.g., the combination product claimed in this invention).

[0159] As used in this article, the term “treatment” means to reduce, alleviate or improve the symptoms of a disease or condition, improve underlying metabolic symptoms, suppress a disease or condition, such as preventing the development of a disease or condition, alleviating a disease or condition, causing the remission of a disease or condition, relieving the condition caused by a disease or condition, or preventing the symptoms of a disease or condition.

[0160] The terms "pharmaceutical-grade carrier" or "pharmaceuticalally acceptable excipient" refer to carriers or excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound.

[0161] The intermediate compounds of the present invention can be prepared by various synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0162] The chemical reactions in the specific embodiments of this invention are carried out in a suitable solvent, which must be suitable for the chemical changes of this invention and the reagents and materials required therefor. To obtain the compounds of this invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0163] The present invention will be described in detail below through embodiments, which are not intended to limit the present invention in any way.

[0164] All solvents used in this invention are commercially available and can be used without further purification.

[0165] The "chemical ratio" of this invention refers to the molar ratio, that is, the molar ratio of the free form compound to the acid.

[0166] In vitro kinase activity inhibition assays showed that the compound shown in formula (A) exhibited better inhibitory activity against PRMT5 and MV4-11 cells compared to the control compound.

[0167] The results of in vivo tumor suppression experiments showed that, compared with the control compound, the compound shown in formula (A) had significant tumor suppression activity, that is, it had a significantly better inhibitory effect on tumor weight and tumor volume.

[0168] In vivo pharmacokinetic studies showed that the compound represented by formula (A) had better oral administration performance, with faster and better absorption.

[0169] Based on one or more of the effects in (1), (2) or (3) above, the salt of compound (A) has a more valuable application prospect.

[0170] The salt of the compound shown in formula (A) or the compound shown in formula (A-1) was obtained for the first time.

[0171] For the first time, salts of the compound shown in formula (A) or the compound shown in formula (A-1) in solid form and salts of the compound shown in formula (A) or the compound shown in formula (A-1) in crystalline form are obtained, facilitating separation, transfer and weighing.

[0172] The salt of the compound shown in formula (A) or the compound shown in formula (A-1) has good solubility.

[0173] For the first time, compounds of formula (B), (B-1), (C), (C-1), or (C-2) were obtained in solid form, exhibiting good properties that facilitate separation, transfer, and weighing.

[0174] The compounds of formula (B), (B-1), (C), (C-1), or (C-2) in solid form can be prepared and isolated with high purity (greater than 95%) and / or yield (greater than 80%).

[0175] The crystalline form of the compound shown in formula (B), the compound shown in formula (B-1), the crystalline form of the compound shown in formula (C), the compound shown in formula (C-1), or the compound shown in formula (C-2) has good crystallinity.

[0176] The crystalline form of the compound of formula (B) and its specific crystal form, and the crystalline form of the compound of formula (C) and its specific crystal form are easy to purify and separate (e.g., by filtration) and are simple to prepare.

[0177] The preferred crystal form possesses good physical and chemical stability and has promising potential for medicinal use. Attached Figure Description

[0178] Figure 1 X-ray powder diffraction pattern of crystal form I of compound (B) in Example 1.

[0179] Figure 2 X-ray powder diffraction pattern of crystal form II of compound (B) in Example 2.

[0180] Figure 3 X-ray powder diffraction pattern of crystal form III of compound (B) in Example 3.

[0181] Figure 4 X-ray powder diffraction pattern of crystal form IV of compound (B) in Example 4.

[0182] Figure 5X-ray powder diffraction pattern of crystal form V of compound of formula (B) in Example 5.

[0183] Figure 6 X-ray powder diffraction pattern of crystal form VI of compound (B) in Example 6.

[0184] Figure 7 X-ray powder diffraction pattern of crystal form VII of compound of formula (B) in Example 7.

[0185] Figure 8 X-ray powder diffraction pattern of crystal form I of compound of formula (C) in Example 8.

[0186] Figure 9 X-ray powder diffraction pattern of crystal form II of compound of formula (C) in Example 9.

[0187] Figure 10 X-ray powder diffraction pattern of crystal form III of compound of formula (C) in Example 10.

[0188] Figure 11 X-ray powder diffraction pattern of crystal form IV of compound of formula (C) in Example 11. Detailed Implementation

[0189] 1. X-ray powder diffractometer (XRPD)

[0190]

[0191] 2. Differential Scanning Calorimeter (DSC)

[0192] Instrument Model TA Discovery 2500 TA Discovery DSC 250 Test sample Examples 7, 9-11, 13-14 Examples 1-6 Sample dosage 1-10mg 2-3mg Sample tray Aluminum disc, pressure cap Aluminum disc, with a lid and perforated Temperature range 25～280℃ 25～250 heating rate 10℃ / min 10℃ / min Protective gas Nitrogen Nitrogen

[0193] 3. Thermogravimetric Analysis (TGA)

[0194] Instrument Model TA Discovery 5500 TA Discovery TGA 55 Test sample Examples 7, 9-11, 13-14 Examples 1-6 Sample dosage 1-10mg 2-3mg Sample tray Aluminum disc, open aluminum disc Temperature range Room temperature ~ 350℃ Room temperature ~ 300℃ heating rate 10℃ / min 10℃ / min Protective gas Nitrogen Nitrogen

[0195] 4. Nuclear Magnetic Resonance Spectroscopy (NMR)

[0196] Instrument model: Bruker 400M MRI scanner (Bruker, GER)

[0197] Contents and test solvents: 1 H-NMR, the test solvent was DMSO-d6.

[0198] 5. Infrared Spectroscopy (IR)

[0199] Testing instrument: PerkinElmer Spectrum 100FT-IR Infrared Spectrometer

[0200] Test method: Weigh 3mg of sample, dilute with KBr and compress into a pellet, then perform the test at room temperature. Specific parameters: Detection range: 4000-400cm². -1 Wavenumber, resolution: 4cm -1 .

[0201] 6. Ion chromatography (IC)

[0202] Detection instrument: ThermoFisher ICS-1100 ion chromatograph

[0203] chromatographic column <![CDATA[Ionpac TM AS18A RFICTM 4×250mm Analytical]]> Injection volume 25μL Flow rate 1.0 mL / min Sample cell temperature / column temperature 35℃ / 35℃ Current 80mA

[0204] 7. High Performance Liquid Chromatography (HPLC)

[0205] Testing instrument: Agilent 1260

[0206]

[0207] 8. Single crystal diffraction

[0208]

[0209] 9. Solubility determination

[0210] FaSSIF: simulates the intestinal fluid in the small intestine of humans in a state of pre-meal hunger; FeSSIF: simulates the intestinal fluid in the small intestine of humans in a state of post-meal satiety; SGF: simulates the gastric fluid in the stomach of humans in a state of hunger when the stomach is empty.

[0211] Test method: Add the sample to be tested to water or biological solvent (4 mL); mix by rotating in a rotary incubator at 37°C (~25 rpm); take about 0.8 mL of solution or suspension at the sampling point (in this invention, the sampling point is 24 hours) into a centrifuge tube and centrifuge; filter the supernatant using a filter membrane, and use the filtrate to test the solubility.

[0212] To better understand the content of this invention, further explanation is provided below with reference to specific embodiments. However, the specific implementation methods are not intended to limit the scope of this invention. For test methods not specified in the following preparation examples, examples, test examples, or experimental examples, conventional methods and conditions should be followed, or the methods should be selected according to the product instructions.

[0213] Preparation Example 1: Preparation of Compound (A)

[0214]

[0215] Preparation of intermediate T-0: In a 100 mL reaction flask, 6-chloro-pyrimidin-4-carboxyl chloride (0.63 g, 3.56 mmol) was dissolved in dichloromethane (10 mL), and triethylamine (0.72 g, 7.12 mmol) was added at 0 °C. Then (S)-1-amino-3-(3,4-dihydroisoquinoline-2(1H)-yl)prop-2-ol (0.66 g, 3.20 mmol) was added. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed by TLC, the reaction mixture was diluted with water (5 mL) and extracted with dichloromethane (15 mL × 2). The combined organic layers were washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent, yielding the residue. The residue was purified by column chromatography (dichloromethane / methanol = 10 / 1, v / v, the same below) to obtain intermediate compound T-0 (yellow oil, 0.68 g, 61.28%).

[0216] Preparation of intermediate T-1: (S)-6-chloro-N-(3-(3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropyl)pyrimidine-4-carboxamide (T-0, 0.28 g, 0.81 mmol) was dissolved in isopropanol (20 mL), followed by the addition of triethylamine (0.25 g, 2.47 mmol) and tert-butyl 4-aminopiperidine-1-carboxylate (0.24 g, 1.2 mmol). The reaction mixture was heated to 85 °C and reacted for 8 h. After the reaction was complete as indicated by TLC, the reaction mixture was concentrated to dryness under reduced pressure and separated by column chromatography (dichloromethane / methanol = 30 / 1) to obtain intermediate compound T-1 (0.36 g, 87.32%).

[0217] Preparation of intermediate T-2: (S)-4-((6-((3-(3,4-dihydroisoquinoline-2(1H)-yl)-2-hydroxypropyl)carbamoyl)pyrimidin-4-yl)amino)piperidin-1-carboxylic acid tert-butyl ester (T-1, 0.36 g, 0.71 mmol) was dissolved in dichloromethane (20 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 h. The solution was concentrated to dryness under reduced pressure to obtain intermediate compound T-2 (0.25 g, 86.38%). LC-MS: m / z 411.54 [M+H] + .

[0218] Preparation of intermediate T-3: 2-O-propionic acid (0.69 g, 7.84 mmol, 1 eq) was dissolved in a mixed solvent of tetrahydrofuran (20 mL) and water (100 mL), and KOH (0.48 g, 8.62 mmol, 1.1 eq) was added. The mixture was stirred until clear, and then methoxyamine hydrochloride (1.31 g, 15.68 mmol, 2.0 eq) was added in portions under ice bath conditions. The reaction was carried out for 2 h, and the mixture was filtered to obtain intermediate compound T-3 (0.91 g, 77.78%). 1 H NMR (600MHz, DMSO-d6): δ1.920(s,3H),3.953(s,3H),12.985(s,1H).

[0219] Preparation of compound A: Intermediate T-2 (0.25 g, 0.61 mmol) was dissolved in DMF (20 mL), followed by the addition of TEA (3 mL, pH > 10), intermediate T-3 (0.085 g, 0.73 mmol), and HATU (0.30 g, 0.789 mmol). The reaction was allowed to proceed at room temperature for 2 h. After the reaction was complete as indicated by TLC, water and ethyl acetate were added, and the mixture was separated. The organic phase was washed once with water and once with saturated brine, then concentrated to dryness and separated by column chromatography (dichloromethane / methanol = 30 / 1) to obtain the target compound (0.22 g, 70.75%). LC-MS: m / z 510.34 [M+H] + . 1 HNMR (600MHz, CD3OD): δ1.459-1.533(m,2H),1.996(s,3H),2.036-2.063(m,2H), 2.645-2.655(m,2H),2.814-2.848(m,2H),2.907-2.926(m,2H),3.014(m,1H),3. 261-3.303(m,1H),3.466-3.550(m,2H),3.710(s,2H),3.909(s,3H),4.018-4.07 4(m,2H),4.210(s,1H),4.399-4.420(d,1H),6.987-7.108(m,5H),8.238(s,1H).

[0220] Example 1: Preparation of compound (B)

[0221] Weigh 0.5 g of the sample from Preparation Example 1, dissolve it in isopropanol (5 mL), add L-malic acid (2 eq), stir at room temperature for about 10 h, filter, and dry at 50 °C ± 5 °C for 8 h to obtain a solid with a yield of 92.5%. Analysis confirmed that it had formed a salt with a base / acid ratio of 1:1. IR (KBr, cm⁻¹) -1):3301.35, 2939.94, 1610.73, 1528.35, 1455.99, 1367.93, 1045.03.

[0222] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal form I) with good crystallization; the spectrum is shown below. Figure 1 The XRPD diffraction peak data are shown in Table 1. DSC-TGA tests were performed on the sample. The DSC plot showed endothermic peaks at 102.15℃ and 113℃, while the TGA plot showed an 8.6522% weight loss between room temperature and 110℃.

[0223] Table 1. XRPD diffraction peak data of the samples obtained in Example 1

[0224] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.205 100.0 13.329 10.3 20.111 6.1 6.534 13.2 18.277 6.5 21.986 7.8 8.532 4.7 19.067 17.8 22.399 5.1

[0225] Note: Peaks with a relative peak intensity > 4.0% are listed in the table.

[0226] Example 2: Preparation of crystal form II of compound (B)

[0227] Weigh 30 mg of the sample obtained in Example 1, add tetrahydrofuran (1.1 mL) to prepare a solution, add n-heptane (0.4 mL) dropwise, stir at room temperature for 7 days, filter, and vacuum dry at 50 °C for 2 hours to obtain a solid. X-ray powder diffraction of the sample showed it to be a crystalline solid (crystal type II) with good crystallinity. The spectrum is shown in [image missing]. Figure 2 The XRPD diffraction peak data are shown in Table 2. DSC-TGA tests were performed on the sample. The DSC plot showed an endothermic peak at 89.8℃ and 103.58℃, while the TGA plot showed a weight loss of 2.6358% between room temperature and 80℃.

[0228] Table 2. XRPD diffraction peak data of crystal form II in Example 2

[0229] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.924 69.1 18.674 100.0 23.294 24.7 6.619 81.0 19.815 79.3 24.012 18.9 12.296 22.7 20.281 41.6 24.384 18.4 13.228 87.2 21.044 13.7 26.532 34.8 13.394 75.3 22.100 45.1 28.798 12.2 16.487 12.9 23.285 25.8

[0230] Note: Peaks with a relative peak intensity >12.0% are listed in the table.

[0231] Example 3: Preparation of Crystal Form III of Compound (B)

[0232] Weigh approximately 50 mg of the sample from Example 1 into a sample vial, add 1.5 mL of acetonitrile, stir for 7 days at room temperature, and filter to obtain a solid. X-ray powder diffraction of the sample showed it to be a crystalline solid (crystal type III) with good crystallinity; the spectrum is shown below. Figure 3The XRPD diffraction peak data are shown in Table 3. DSC-TGA tests were performed on the sample. The DSC plot showed endothermic peaks at 70.82℃ and 117.93℃, while the TGA plot showed a weight loss of 3.0001% between room temperature and 75℃.

[0233] Table 3. XRPD diffraction peak data of crystal form III in Example 3

[0234]

[0235]

[0236] Note: Peaks with a relative peak intensity >10.0% are listed in the table.

[0237] Example 4: Preparation of crystal form IV of compound (B)

[0238] Weigh approximately 50 mg of the sample from Example 1 into a sample vial, add 1.5 mL of a methanol / methyl tert-butyl ether mixture (v / v = 1 / 3), stir at room temperature for 7 days, and filter to obtain a solid. X-ray powder diffraction of the sample showed it to be a crystalline solid (crystal form IV) with good crystallinity; the spectrum is shown below. Figure 4 The XRPD diffraction peak data are shown in Table 4. DSC-TGA tests were performed on the sample. The DSC plot showed two endothermic peaks at 77.91℃ and 113.27℃, while the TGA plot showed a weight loss of 2.6271% between room temperature and 60℃.

[0239] Table 4. XRPD diffraction peak data of crystal form IV in Example 4

[0240] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.446 100.0 13.870 27.3 18.512 12.0 7.598 28.6 15.194 11.9 20.630 39.3 8.890 27.4 17.115 11.1 21.340 8.7 12.143 12.0 17.799 16.6

[0241] Note: Peaks with a relative peak intensity > 8% are listed in the table.

[0242] Example 5: Preparation of crystal form V of compound (B)

[0243] Weigh approximately 50 mg of the sample from Example 1 into a sample vial, add 1.5 mL of acetone, stir for 7 days at room temperature, and filter to obtain a solid. X-ray powder diffraction of the sample showed it to be a crystalline solid (crystal form V) with good crystallinity; the spectrum is shown below. Figure 5 The XRPD diffraction peak data are shown in Table 5. DSC-TGA tests were performed on the sample. The DSC plot showed two endothermic peaks at 68.38℃ and 104.69℃, while the TGA plot showed a weight loss of 3.6041% between room temperature and 75℃.

[0244] Table 5. XRPD diffraction peak data of crystal form V in Example 5

[0245] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.971 89.9 16.452 11.9 22.789 15.9 6.573 30.4 16.700 20.2 23.233 26.9 6.788 46.4 18.645 100.0 24.259 26.1 12.418 17.7 19.090 42.2 24.962 22.5 12.796 24.3 19.592 62.5 25.708 15.2 13.070 36.0 20.200 46.1 28.640 15.4 13.646 60.7 20.407 19.5 14.350 16.2 21.763 15.5

[0246] Note: Peaks with a relative peak intensity >10.0% are listed in the table.

[0247] Example 6: Preparation of crystal form VI of compound (B)

[0248] Weigh an appropriate amount of crystal form IV obtained in Example 4 and vacuum dry it at 50°C for 2 days to obtain a solid. X-ray powder diffraction of the sample showed it to be a crystalline solid (crystal form VI) with good crystallinity; the spectrum is shown below. Figure 6 The XRPD diffraction peak data are shown in Table 6. DSC-TGA tests were performed on the sample. The DSC plot showed an endothermic peak at 113.29℃, and the TGA plot showed a weight loss of 0.34% between room temperature and 120℃.

[0249] Table 6. XRPD diffraction peak data of crystal form VI in Example 6

[0250] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.520 95.3 13.284 56.2 19.326 11.3 6.956 100.0 13.904 21.8 20.157 79.8 9.026 85.3 15.845 27.2 21.613 33.9 12.921 74.5 16.931 13.3 25.367 11.7

[0251] Note: Peaks with a relative peak intensity >10.0% are listed in the table.

[0252] Example 7: Preparation of crystal form VII of compound (B)

[0253] The sample (501.3 mg) obtained in Preparation Example 1 was weighed into a 20 mL reaction flask, and ethanol (12 mL) was added to prepare a solution. L-malic acid (131.6 mg) was added, and the mixture was magnetically stirred at room temperature for 8 days. The solid was separated by centrifugation and vacuum dried at room temperature for 1 day, with a purity of 98.22%. NMR analysis confirmed that the sample had formed a salt with a base / acid ratio of approximately 1:1.5.

[0254] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal type VII) with good crystallinity; the spectrum is shown below. Figure 7 The XRPD diffraction peak data are shown in Table 7. DSC-TGA tests were performed on the sample. The DSC plot showed endothermic peaks at 92.4℃ and 197.3℃, while the TGA plot showed a 7.88% weight loss between room temperature and 110℃.

[0255] Table 7. XRPD diffraction peak data for crystal form VII in Example 7

[0256] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.22 90.01 8.92 24.47 19.05 100.00 4.44 86.27 12.56 19.47 19.98 94.97 6.58 66.12 13.24 80.99 21.09 19.27 6.91 67.56 15.12 15.06 21.70 25.80

[0257] Note: Peaks with a relative peak intensity >15.0% are listed in the table.

[0258] Example 8: Preparation of Crystal Form I of Compound (C)

[0259] Weigh 0.5 g of the sample from Preparation Example 1, dissolve it in isopropanol (5 mL), add oxalic acid (2 eq), stir at room temperature for about 10 h, filter, and dry at 50 °C ± 5 °C for 6 h to obtain a solid with a yield of 90%. Analysis confirmed that it had formed a salt. IR (KBr, cm⁻¹) -1 ):3319.96, 2936.96, 1615.04, 1526.12, 1455.78, 1368.16, 1046.84.

[0260] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal form I) with good crystallization; the spectrum is shown below. Figure 8 The XRPD diffraction peak data are shown in Table 8.

[0261] Table 8. XRPD diffraction peak data of crystal form I in Example 8

[0262] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.734 100.0 16.327 27.7 21.213 20.5 5.323 19.9 16.744 17.3 22.708 13.9 7.136 12.4 17.373 32.4 23.244 20.8 10.714 18.1 18.365 12.9 23.696 17.6 14.082 9.9 19.569 18.3 16.128 26.9 20.122 20.0

[0263] Note: Peaks with a relative peak intensity > 9.0% are listed in the table.

[0264] Example 9: Preparation of Crystal Form II of Compound (C)

[0265] 6.4 mg of oxalic acid dihydrate was weighed and added to a vial. An ethanol solution (40 mg / mL, 0.5 mL) of the sample from Preparation Example 1 was added. The mixture was magnetically stirred at room temperature for 3 days. The solid was then centrifuged and vacuum dried at room temperature for 1 day, yielding a purity of 98.56%. Testing confirmed that the sample had formed a salt with an alkali / acid ratio of approximately 1:1.5.

[0266] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal type II) with good crystallinity; the spectrum is shown below. Figure 9 The XRPD diffraction peak data are shown in Table 9. DSC-TGA tests were performed on the sample. The DSC plot showed an endothermic peak at 162.6℃, and the TGA plot showed a 3.20% weight loss between room temperature and 160℃.

[0267] Table 9. XRPD diffraction peak data for crystal form II in Example 9

[0268] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.55 100.00 18.44 13.96 24.50 16.74 4.77 39.33 19.39 14.75 24.97 15.59 12.65 13.49 19.58 10.27 27.33 9.99 13.02 60.61 21.57 13.95

[0269] Note: Peaks with a relative peak intensity > 8.0% are listed in the table.

[0270] Example 10: Preparation of Crystal Form III of Compound (C)

[0271] 6.3 mg of oxalic acid dihydrate was weighed and added to a vial. 0.5 mL of 40 mg / mL 2-methyltetrahydrofuran solution (prepared in Example 1) was added. The mixture was magnetically stirred at room temperature for 3 days. The solid was then centrifuged and vacuum dried at room temperature for 1 day. Analysis confirmed that a salt had formed, with an alkali / acid ratio of approximately 1:1.

[0272] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal type III) with good crystallinity; the spectrum is shown below. Figure 10 The XRPD diffraction peak data are shown in Table 10. DSC-TGA tests were performed on the sample. The DSC plot showed endothermic peaks at 139.9℃ and 204.9℃, while the TGA plot showed a 6.20% weight loss between room temperature and 150℃.

[0273] Table 10. XRPD diffraction peak data of crystal form III in Example 10

[0274] Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % Peak position (2θ)° Relative strength % 4.55 100.00 16.71 14.80 20.41 8.49 4.83 54.70 17.56 9.63 21.07 9.78 13.65 6.13 18.13 7.89 21.61 9.97 14.01 11.46 18.72 16.95 23.28 13.37 16.41 9.38 19.36 16.21

[0275] Note: Peaks with a relative peak intensity > 5.0% are listed in the table.

[0276] Example 11: Preparation of crystal form IV of compound (C)

[0277] Oxalic acid dihydrate (123.9 mg) and sample from Preparation Example 1 (503.0 mg) were weighed and added to a reaction flask. Ethanol (10 mL) was added, and the mixture was magnetically stirred at room temperature for 2 days. The solid was then separated by centrifugation and vacuum dried at room temperature for 3 days, with a purity of 98.96%. Testing confirmed that the solid had formed a salt with an alkali / acid ratio of approximately 1:1.

[0278] X-ray powder diffraction of the sample revealed it to be a crystalline solid (crystal type IV) with good crystallinity; the spectrum is shown below. Figure 11 The XRPD diffraction peak data are shown in Table 11. DSC-TGA tests were performed on the sample. The DSC plot showed an endothermic peak at 137.4℃, and the TGA plot showed a weight loss of 4.26% between room temperature and 150℃.

[0279] Table 11. XRPD diffraction peak data of crystal form IV in Example 11

[0280]

[0281]

[0282] Note: Peaks with a relative peak intensity > 2.0% are listed in the table.

[0283] Example 12: Preparation of crystal form V of compound (C)

[0284] Weigh 41 mg of the sample from Preparation Example 1 and 12.9 mg of oxalic acid dihydrate into a glass bottle, add 0.5 mL of acetone / water mixed solvent (v / v = 1 / 1), heat to 80 °C and stir for 12 h, then slowly cool to room temperature to obtain a single crystal of compound (C) (acid-base ratio: 1:1). Using a Cu-Kα light source, the obtained single crystal belongs to the triclinic crystal system, space group P1, and its unit cell parameters are: { α=99.1280(10)°, β=90.1780(10)°, γ=95.1340(10)°, }

[0285] Example 13: Preparation of organic acid addition salts of other compounds of formula (A)

[0286] The sample from Preparation Example 1 was dissolved in the solvents shown in the table below to prepare a solution with a concentration of 40 mg / mL. Different acids were weighed and added to a vial, and then the prepared sample solution from Preparation Example 1 (0.5 mL) was added. The specific experimental method is shown in the table below. The solid was separated by centrifugation and vacuum dried at room temperature for 1 day.

[0287] NMR or HPLC analysis showed that the sample from Preparation Example 1 could form salts with all the acids listed in the table below, with a base / acid ratio of 1:1, and solids could be separated from them. Further, X-ray powder diffraction was performed on the samples, and the specific results are detailed in the table below:

[0288] Table 12. Preparation and Results of Addition Salts of Other Organic Acids

[0289]

[0290] Example 14: Preparation of inorganic acid addition salt of compound (A)

[0291] The following salts were prepared using the same method as in Example 13, and the specific results are shown in the table below:

[0292] Table 13. Preparation and results of inorganic acid addition salts

[0293]

[0294]

[0295] Test Example 1: Solid stability experiment of different crystal forms of compound (B) under different humidity conditions

[0296] Weigh appropriate amounts of samples from Example 2 (crystal form II of compound (B)), Example 3 (crystal form III of compound (B)) and Example 4 (crystal form IV of compound (B)) into vials, and place them at 23℃ / 33%RH and 25℃ / 60%RH for 7 days respectively. Take samples for X-ray powder diffraction to examine the stability of the samples under different conditions. The results are shown in the table below.

[0297] Table 14. Experimental results of solid stability of samples with different crystal forms

[0298]

[0299] Conclusion: The crystal forms of crystal form II in Example 2, crystal form III in Example 3, and crystal form IV in Example 4 remained unchanged when placed under different humidity conditions.

[0300] Test Example 2: Solid stability experiment of different crystal forms of compound (B)

[0301] Weigh appropriate amounts of samples from Example 2 (crystal form II of compound (B)) and Example 5 (crystal form V of compound (B)) into vials and place them under high temperature (60°C, sealed) conditions for 7 days. Take samples for X-ray powder diffraction to examine the stability of the samples under different conditions. The results are shown in the table below.

[0302] Table 15. Experimental results on the solid stability of different crystal forms of compound (B)

[0303]

[0304] Conclusion: Crystal form II of Example 2 and crystal form V of Example 5 can maintain crystal form stability after being placed under high temperature conditions for 7 days.

[0305] Test Example 3: Long-term (3-month) stability test of crystal form I of compound (B)

[0306] Weigh an appropriate amount of the sample from Example 1 (crystal form I of compound (B)) into a vial and conduct a long-term stability study. The results are shown in the table below.

[0307] Table 16. Results of long-term (3-month) stability test of crystal form I of compound (B)

[0308]

[0309] Conclusion: Crystal form I of Example 1 maintained essentially the same purity and crystal form during a long-term experiment lasting 3 months.

[0310] Test Example 4: Solubility Test

[0311] The salts of the compounds of formula (A) obtained in the above examples have good solubility in water and biological solvents, as shown in the following representative examples.

[0312] The solubility of Example 7 (compound of formula (B), crystal form VII), Example 10 (compound of formula (C), crystal form) and the free sample (preparation example 1) in water, SGF and FassIF was tested at 37°C for 24 hours. The results are shown in the table below.

[0313]

[0314] Note: ① Solubility is expressed in free state; ②> indicates that all the added sample dissolved and no further sample was added.

[0315] The crystal form VII of compound (B) and the crystal form IV of compound (C) showed significantly higher solubility in water and FassIF medium than the free alkali sample obtained in Preparation Example 1.

[0316] Experimental Example 1: In vitro efficacy test

[0317] 1. Enzymatic experimental methods

[0318] IC of test substances was detected on a PRMT5 using the radioisotope FlashPlate technique. 50 .

[0319] The test compounds were dissolved in dimethyl sulfoxide (DMSO), added to Echo 384-well plates, and diluted to the required concentration. Using an Echo 550 instrument, the test compounds were transferred from the diluted Echo 384-well plates to 384-well reaction plates. Control and blank wells were transferred to DMSO. PRMT5 was added to 1x reaction buffer (1x reaction buffer included 10mM Tris-HCl; pH 8.0; 0.01% Tween-20; 1mM DTT) to prepare a 1.67x enzyme solution (enzyme concentration of 5nM). The peptide substrate and [ 3Add 1x H-SAM to the reaction buffer to prepare a 2.5x substrate solution (final substrate concentrations of 100 nM and 250 nM). Add 15 μL / well of 1.67x enzyme solution to each well of a 384-well plate. For blank wells, replace the enzyme solution with 15 μL of 1x reaction buffer. Centrifuge at 1000 rpm for 1 min and incubate at room temperature for 15 min. Add 10 μL of 2.5x substrate solution to each well of the 384-well plate. Centrifuge at 1000 rpm for 1 min. Incubate at 25°C for 60 min. Stop the reaction by adding 5 μL of stop solution (125 μM cold SAM solution) to each well of the 384-well plate. Transfer 25 μL from each well of the plate to a Flashplate and incubate at room temperature for 1 h. Then wash the Flashplate three times with 0.1% Tween-20 solution. Read the data using a MicroBeta 2. Convert the data into inhibition rate data. Inhibition rate = (conversion rate) 对照孔 -conversion rate 化合物孔 ) / (conversion rate) 对照孔 -conversion rate 空白孔 ()×100%. Fit IC using XLFit 5.4.0.8. 50 Value. The fitting formula is Y = Bottom + (Top - Bottom) / (1 + (IC)). 50 / X)^HillSlope).

[0320] 2. Cellular Experimental Methods

[0321] The complete culture medium used for culturing human acute monocytic leukemia cells MV4-11 (Shanghai Cell Bank) was IMDM (Cat NO. 12440-053, Gibco) supplemented with 10% serum-FBS (Cat NO. SA311.02, Cellmax). Cells were cultured at 37°C in a 5% CO2 incubator. Experimental reagents included dimethyl sulfoxide (Tianjin Kemei Chemical Reagent Co., Ltd.) and MTT (THIAZOLYL BLUE TETRAZOLIUM BROMIDE, CAS. NO. 298-93-1, VWR). The control substance GSK3326595 was obtained either in-house or commercially available. The test samples were sealed and stored at 4°C.

[0322] Using dimethyl sulfoxide as the solvent, the test substance was fully dissolved to prepare a solution with a concentration of 5 × 10⁻⁶. -2 Prepare a mol / L stock solution and store it at -20°C. Use complete culture medium as a diluent to serially dilute the test substance to different concentrations. Add 100 μL / well (2 × 10⁻⁶) to each well of a 96-well plate. 3Cells (cells / well) were added to a complete culture medium suspension of human acute monocytic leukemia cells MV4-11. Then, different concentrations of the test agent (100 μL / well) were added, with eight concentrations for each test agent and three replicates for each concentration. Cells were incubated at 37°C in a 5% CO2 incubator. On day six, MTT (20 μL / well) was added, and the cells were incubated at 37°C in a 5% CO2 incubator for 4 hours. The supernatant was discarded, and dimethyl sulfoxide (150 μL / well) was added. The cells were vortexed and the OD value was measured at 550 nm using a microplate reader. Wells containing only cell suspension without the test agent were control wells, and wells containing only complete culture medium were blank wells. The cell growth inhibition rate was calculated using the following formula:

[0323] Inhibition rate = (OD value) 对照孔 -OD value 给药孔 ) / (OD value) 对照孔 -OD value 空白孔 The half-maximum inhibitory concentration (IC50) was calculated using SPSS software based on the inhibition rate at each concentration (increased by 100%). 50 The values ​​are shown in Table 18.

[0324] Table 17. Enzyme efficacy test data

[0325] Test compound <![CDATA[Enzymology (IC 50 , nM)]]> GSK3326595 47 Compound of formula (A) 31

[0326] Table 18. Cellular efficacy test data

[0327] Test compound <![CDATA[MV4-11 MTT(IC 50 ,nM)]]> GSK3326595 6.683 Compound of formula (A) 6.767

[0328] Enzymatic and cellular screening results showed that, compared with the positive control drug GSK3326595 (compound 208 in WO2014100719), the compound of formula (A) of the present invention exhibited better inhibitory activity at both the enzyme and cellular levels.

[0329] Example 2: In vivo efficacy test

[0330] Female NOD-SCID mice, SPF grade, 4-5 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Mice were intraperitoneally injected with cyclophosphamide at a dose of 100 mg / kg one day before cell inoculation. Human acute monocytic leukemia cells MV4-11 (1 x 10⁻⁶ cells) were subcutaneously inoculated into the axilla of the forelimbs of mice. 7 A subcutaneous xenograft model was established using 0.1 ml / tumor. The tumor volume was increased to approximately 110 mm². 3On day 10 post-inoculation, mice were divided into two groups of five mice each, based on tumor volume: a solvent control group (2% DMSO + 98% (0.2 g / mL) hydroxypropyl β-cyclodextrin) and a test drug group. The test drug group received a dose of 100 mg / kg, a volume of 10 mL / kg, and was administered twice daily (BID). Tumor diameter was measured twice weekly, and data were recorded. The treatment lasted for 11 consecutive days. At the end of the experiment, the tumors were removed and weighed.

[0331] The weight gain rate, tumor volume, and tumor weight inhibition rate are calculated using the formula: Weight gain rate = X (Wi-W0) / X W0 ×100%, where W i W represents the weight of a mouse in each experimental group on day n, and W0 represents the weight of a mouse in each experimental group at the start of drug administration; tumor volume (V) = 1 / 2 × a × b 2 Where a and b represent the long and short diameters of the tumor, respectively; d0 is before administration, d9 is the 9th day after administration, relative tumor volume (RTV) = tumor volume on d9 / tumor volume on d0; tumor weight inhibition rate = (tumor weight) / tumor volume on d0. 溶媒对照组 -tumor weight 受试药物组 ) / tumor weight 溶媒对照组 ×100%, the results are shown in Table 19.

[0332] Table 19. In vivo test data for MV4-11

[0333]

[0334]

[0335] Note: Compared with the solvent control group, *p<0.05, **p<0.01, ***p<0.001; compared with the control drug GSK3326595, ##p<0.01, ###p<0.001.

[0336] The in vivo efficacy test results of MV4-11 showed that the compound of formula (A) of the present invention has significant antitumor activity, and its inhibitory effect on tumor volume and tumor weight is significantly better than that of the control drug GSK3326595.

[0337] Experimental Example 3: Pharmacokinetic Study

[0338] Male SD rats (180-200g, Beijing Vital River Laboratory Animal Technology Co., Ltd.) were administered the test compound (solvent: 2% DMSO + 98% (0.2g / mL) hydroxypropyl β-cyclodextrin, administration volume 5mL / kg) by gavage. Blood samples were collected from the rats' orbital sinuses at different time points after administration (0.25, 0.5, 1, 2, 4, 8, 24h). The collected whole blood was anticoagulated with heparin sodium and centrifuged at 3000g to obtain rat plasma samples. Methanol protein precipitation was used, and the drug concentration in rat plasma after administration was determined by HPLC-MS / MS. Drug-time curves were plotted, and pharmacokinetic parameters were calculated. The pharmacokinetic behavior of the compound in rats after administration was described by non-compartmental model statistical moment parameters. The results are shown in Table 20.

[0339] Table 20. Pharmacokinetic data of SD rats

[0340]

[0341] Pharmacokinetic studies showed that, compared with the positive control drug GSK3326595, the compound of formula (A) of this invention was better absorbed in rats, with a shorter time to peak concentration, higher peak concentration, and significantly increased in vivo exposure.

[0342] Although the foregoing invention has been described in considerable detail by way of illustration and example for the purpose of clear understanding, it will be apparent from the teachings of the invention that those skilled in the art may make certain changes and modifications thereto without departing from the spirit or scope of the appended claims.

Claims

1. A compound, said compound being an inorganic acid addition salt or an organic acid addition salt of the compound represented by formula (A), ， in, The inorganic acid addition salt is a sulfate or a phosphate; The organic acid addition salts are selected from malate, oxalate, succinate, tartrate, adipate, citrate, gluconate, maleate, fumarate, lactate, and gentianate.

2. The compound according to claim 1, wherein, The organic acid addition salt is selected from malate, oxalate, succinate, L-tartrate, L-lactate, adipate, citrate and gluconate.

3. The compound according to claim 1, wherein, The organic acid addition salt is selected from L-malate and oxalate.

4. The compound according to any one of claims 1 to 3, wherein, The chemical ratio of the compound shown in formula (A) to organic or inorganic acid molecules is 1:1 to 1.

5.

5. The compound according to any one of claims 1 to 3, wherein, The chemical ratio of the compound shown in formula (A) to organic or inorganic acid molecules is 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4 or 1:1.

5.

6. The compound according to any one of claims 1 to 3, wherein, The chemical ratio of the compound shown in formula (A) to organic or inorganic acid molecules is 1:

1.

7. A compound, said compound being the compound represented by formula (B), ， in, n is selected from 1 to 1.

5.

8. The compound according to claim 7, wherein, n is 1, 1.1, 1.2, 1.3, 1.4 or 1.

5.

9. The compound according to claim 7, wherein, n is 1 or 1.

5.

10. The compound according to claim 7, wherein, The compound is in solid form and was prepared by KBr pelleting. Its infrared spectrum includes a characteristic peak at the following location: 3301±4 cm⁻¹. -1 2940±4 cm -1 1611±4 cm -1 1528±4 cm -1 1456±4 cm -1 1368±4 cm -1 1045±4 cm -1 .

11. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form I, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.2±0.2°, 6.5±0.2°, 13.3±0.2°, 19.1±0.2°, 20.1±0.2°, and 22.0±0.2°.

12. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form I, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.2±0.2°, 6.5±0.2°, 13.3±0.2°, 18.3±0.2°, 19.1±0.2°, 20.1±0.2°, and 22.0±0.2°.

13. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form I, and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 1.

14. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form II as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.9±0.2°, 6.6±0.2°, 13.2±0.2°, 18.7±0.2°, and 19.8±0.2°.

15. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form II as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.9±0.2°, 6.6±0.2°, 13.2±0.2°, 18.7±0.2°, 19.8±0.2°, 22.1±0.2°, and 26.5±0.2°.

16. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form II as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.9±0.2°, 6.6±0.2°, 13.2±0.2°, 18.7±0.2°, 19.8±0.2°, 22.1±0.2°, 23.3±0.2°, and 26.5±0.2°.

17. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form II as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.9±0.2°, 6.6±0.2°, 13.2±0.2°, 18.7±0.2°, 19.8±0.2°, 20.3±0.2°, 22.1±0.2°, 23.3±0.2°, and 26.5±0.2°.

18. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form II as shown in formula (B), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 2.

19. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form III as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.3±0.2°, 6.9±0.2°, 8.8±0.2°, 20.3±0.2°, and 21.2±0.2°.

20. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form III as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.3±0.2°, 6.9±0.2°, 8.8±0.2°, 12.8±0.2°, 13.3±0.2°, 20.3±0.2°, and 21.2±0.2°.

21. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form III as shown in formula (B), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 3.

22. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form IV as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.4±0.2°, 7.6±0.2°, 8.9±0.2°, 13.9±0.2°, and 20.6±0.2°.

23. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form IV as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.4±0.2°, 7.6±0.2°, 8.9±0.2°, 12.1±0.2°, 13.9±0.2°, 15.2±0.2°, and 20.6±0.2°.

24. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form IV as shown in formula (B), and the X-ray powder diffraction pattern of the compound using Cu-Kα radiation has characteristic diffraction peaks at the following 2θ angles: 4.4±0.2°, 7.6±0.2°, 8.9±0.2°, 12.1±0.2°, 13.9±0.2°, 15.2±0.2°, 17.8±0.2°, 18.5±0.2°, and 20.6±0.2°.

25. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form IV as shown in formula (B), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 4.

26. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form V, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 5.0±0.2°, 13.6±0.2°, 18.6±0.2°, 19.6±0.2°, and 20.2±0.2°.

27. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form V, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 5.0±0.2°, 6.8±0.2°, 13.1±0.2°, 13.6±0.2°, 18.6±0.2°, 19.6±0.2°, and 20.2±0.2°.

28. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form V, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 5.0±0.2°, 6.8±0.2°, 13.1±0.2°, 13.6±0.2°, 16.7±0.2°, 18.6±0.2°, 19.6±0.2°, 20.2±0.2°, and 24.3±0.2°.

29. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form V, and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, exhibits characteristic diffraction peaks at the following 2θ angles: 5.0±0.2°, 6.8±0.2°, 13.1±0.2°, 13.6±0.2°, 16.7±0.2°, 18.6±0.2°, 19.6±0.2°, 20.2±0.2°, 23.2±0.2°, 24.3±0.2°, and 25.0±0.2°.

30. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form V, and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 5.

31. The compound according to any one of claims 7 to 10, wherein, The compound is of the form VI shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.5±0.2°, 7.0±0.2°, 9.0±0.2°, 12.9±0.2°, 20.2±0.2°, and 21.6±0.2°.

32. The compound according to any one of claims 7 to 10, wherein, The compound is of the form VI shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.5±0.2°, 7.0±0.2°, 9.0±0.2°, 12.9±0.2°, 13.3±0.2°, 15.8±0.2°, 20.2±0.2°, and 21.6±0.2°.

33. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form VI as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, exhibits characteristic diffraction peaks at the following 2θ angles: 4.5±0.2°, 7.0±0.2°, 9.0±0.2°, 12.9±0.2°, 13.3±0.2°, 13.9±0.2°, 15.8±0.2°, 20.2±0.2°, and 21.6±0.2°.

34. The compound according to any one of claims 7 to 10, wherein, The compound is of formula (B) in crystal form VI, and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 6.

35. The compound according to any one of claims 7 to 10, wherein, The compound is of the form VII of formula (B) and is subjected to Cu-Kα radiation. The X-ray powder diffraction pattern of the compound subjected to Cu-Kα radiation has characteristic diffraction peaks at the following 2θ angles: 4.3±0.2°, 6.9±0.2°, 13.2±0.2°, 19.1±0.2°, and 20.0±0.2°.

36. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form VII as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.3±0.2°, 6.9±0.2°, 8.9±0.2°, 13.2±0.2°, 19.1±0.2°, 20.0±0.2°, and 21.7±0.2°.

37. The compound according to any one of claims 7 to 10, wherein, The compound is of crystal form VII as shown in formula (B), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.3±0.2°, 6.9±0.2°, 8.9±0.2°, 13.2±0.2°, 15.1±0.2°, 19.1±0.2°, 20.0±0.2°, 21.1±0.2°, and 21.7±0.2°.

38. The compound according to any one of claims 7 to 10, wherein, The compound is of the form VII shown in formula (B) and, using Cu-Kα radiation, has an X-ray powder diffraction pattern substantially as shown in Figure 7.

39. A compound, said compound being the compound represented by formula (C), ， in, n is selected from 1 to 1.

5.

40. The compound according to claim 39, wherein, n is 1, 1.1, 1.2, 1.3, 1.4 or 1.

5.

41. The compound according to claim 39, wherein, n is 1 or 1.

5.

42. The compound according to claim 39, wherein, The compound is in solid form and was prepared using the KBr pellet method. The infrared spectrum of the compound includes a characteristic peak at the following position: 3320±4 cm⁻¹. -1 2937±4 cm -1 1615±4 cm -1 1526±4 cm -1 1456±4 cm -1 1368±4 cm -1 1047±4 cm -1 .

43. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form I as shown in formula (C), and its X-ray powder diffraction pattern, obtained by Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.7±0.2°, 7.1±0.2°, 10.7±0.2°, 17.4±0.2°, 21.2±0.2°.

44. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form I as shown in formula (C), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.7±0.2°, 7.1±0.2°, 10.7±0.2°, 16.1±0.2°, 17.4±0.2°, 20.1±0.2°, and 21.2±0.2°.

45. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form I as shown in formula (C), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, exhibits characteristic diffraction peaks at the following 2θ angles: 4.7±0.2°, 7.1±0.2°, 10.7±0.2°, 16.1±0.2°, 17.4±0.2°, 19.6±0.2°, 20.1±0.2°, 21.2±0.2°, and 23.2±0.2°.

46. ​​The compound according to any one of claims 39 to 42, wherein, The compound is of formula (C) in crystal form I, and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 8.

47. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form II as shown in formula (C), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.6±0.2°, 13.0±0.2°, 21.6±0.2°, 24.5±0.2°, and 25.0±0.2°.

48. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form II as shown in formula (C), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.6±0.2°, 4.8±0.2°, 13.0±0.2°, 18.4±0.2°, 19.4±0.2°, 21.6±0.2°, 24.5±0.2°, and 25.0±0.2°.

49. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form II as shown in formula (C), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 9.

50. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form III as shown in formula (C), and the X-ray powder diffraction pattern of the compound, when subjected to Cu-Kα radiation, has characteristic diffraction peaks at the following 2θ angles: 4.6±0.2°, 4.8±0.2°, 14.0±0.2°, 16.7±0.2°, 18.7±0.2°, 19.4±0.2°, and 23.3±0.2°.

51. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form III as shown in formula (C), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 10.

52. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form IV as shown in formula (C), and the X-ray powder diffraction pattern of the compound under Cu-Kα radiation has characteristic diffraction peaks at the following 2θ angles: 4.6±0.2°, 13.7±0.2°, 19.5±0.2°, 20.0±0.2°, and 22.9±0.2°.

53. The compound according to any one of claims 39 to 42, wherein, The compound is of crystal form IV as shown in formula (C), and using Cu-Kα radiation, the compound has an X-ray powder diffraction pattern substantially as shown in Figure 11.

54. The compound according to any one of claims 39 to 42, wherein, The compound is of formula (C) in crystal form V, and the single crystal obtained by Cu-Kα irradiation belongs to the triclinic crystal system. P Space group 1, its unit cell parameters are: a =5.55690 (10) Å, b =16.9102(2) Å, c =18.9473 (2) Å, α =99.1280(10)°, β =90.1780(10)°, γ =95.1340(10)°, V =1750.57 (4) Å 3 .

55. A crystalline composition comprising one or more of the compounds of formula (B) as claimed in any one of claims 7 to 38.

56. A crystalline composition comprising one or more compounds of formula (C) as claimed in any one of claims 39 to 54.

57. A pharmaceutical composition comprising any one of claims 1 to 54 or the crystalline composition of claim 55 or 56.

58. The pharmaceutical composition according to claim 57, wherein, The pharmaceutical composition also includes a pharmaceutically acceptable carrier.

59. The use of the compound according to any one of claims 1 to 54, the crystalline composition according to claim 55 or 56, or the pharmaceutical composition according to claim 57 or 58 in the preparation of a medicament, wherein, The drug is used to prevent and / or treat diseases mediated at least in part by PRMT5.

60. The use of the compound according to any one of claims 1 to 54, the crystalline composition according to claim 55 or 56, or the pharmaceutical composition according to claim 57 or 58 in the preparation of a medicament for the prevention and / or treatment of proliferative diseases.

61. The application according to claim 60, wherein, The aforementioned proliferative disease is a tumor.

62. The application according to claim 61, wherein, The tumor is either a hematologic malignancy or a solid tumor.

63. The application according to claim 61, wherein, The tumor is a malignant hematologic malignancy or an advanced solid tumor.

64. The application according to claim 61, wherein, The tumor is a relapsed or refractory hematologic malignancy or an advanced malignant solid tumor.

65. The application according to claim 61, wherein, The tumor or cancer mentioned is selected from lung cancer, bone cancer, stomach cancer, pancreatic cancer, adenoid cystic carcinoma, skin cancer, head and neck cancer, uterine cancer, ovarian cancer, testicular cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, brain cancer, pituitary adenoma, melanoma, epidermoid carcinoma, and chronic and acute leukemia.

66. The application according to claim 65, wherein, The acute leukemia mentioned is acute myeloid leukemia.

67. A method for preparing the compound according to any one of claims 1 to 6, comprising reacting the compound of formula (A) with an acid in a reaction solvent, and separating the compound to obtain the compound. 。 68. The preparation method according to claim 67, wherein, The reaction solvent is selected from one or a combination of two of the following: alcohols, esters, nitriles, ketones, water, alkane solvents, ether solvents, or heterocyclic alkane solvents.

69. The preparation method according to claim 67, wherein, The reaction solvent is one or a combination of two of the following: ROH, RCOOR1, RCN, RCOOR1, water, ROR1, RH, or heterocyclic alkane solvents, wherein R and R1 are each independently selected from C 1-6 Straight-chain or branched alkyl groups.

70. The preparation method according to claim 69, wherein, The reaction solvent is wherein R and R1 are each independently selected from C. 1-4 Straight-chain or branched alkyl groups.

71. The preparation method according to claim 67, wherein, The reaction solvent is selected from one or a combination of two of the following: isopropanol, methanol, ethanol, ethyl acetate, acetone, butanone, acetonitrile, water, tetrahydrofuran, n-heptane, and 2-methyltetrahydrofuran.

72. The preparation method according to claim 67, wherein, The reaction temperature ranges from 0°C to 90°C.

73. The preparation method according to claim 67, wherein, The reaction temperature is from 5℃ to 80℃.

74. The preparation method according to claim 67, wherein, The reaction temperature is between 20℃ and 60℃.

75. The preparation method according to claim 67, wherein, The reaction temperature is from room temperature to 50°C.

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