A pharmaceutically acceptable salt of an indazole derivative, a crystalline form thereof, and a method of preparing the same

Abstract

This invention provides a pharmaceutically acceptable salt, a crystalline form, and a method for preparing an indazole derivative. Specifically, it relates to a pharmaceutically acceptable salt crystal form of the compound shown in formula (I) and a method for preparing the same. The provided crystalline form of the pharmaceutically acceptable salt of the compound of formula (I) exhibits good stability and is better suited for clinical treatment.

Description

[0001] This application claims priority to Chinese patent application 202011510169.3, filed on 2020 / 12 / 18. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This disclosure relates to a pharmaceutically acceptable salt, crystalline form, and preparation method of an indazole derivative, and belongs to the pharmaceutical field. Background Technology

[0003] Breast cancer is one of the most common malignant tumors in women. According to GLOBALCAN statistics in 2012 (CACANCER J CLIN 2015; 65:87–108), there are approximately 1.7 million new cancer cases and 520,000 deaths worldwide each year, making it the leading cause of cancer death among women in both incidence and mortality. The 2017 "China Cancer Registry Annual Report" released by the National Cancer Center shows that breast cancer ranks first in incidence among malignant tumors in women, with approximately 279,000 new cases annually, increasing at a rate of about 2% per year.

[0004] Approximately 70% of breast cancer patients have estrogen receptor (ER)-positive breast cancer, and endocrine therapy plays a crucial role in the treatment of this group. Endocrine therapy is mainly divided into three categories: aromatase inhibitors (AIs), which inhibit the conversion of androgens to estrogens and lower estrogen levels in the body; selective estrogen receptor modulators (SERMs), which antagonize estrogen receptor activity; and selective estrogen receptor degraders (SERDs), which not only antagonize estrogen receptor activity but also promote receptor degradation (Pharmacol Ther. 2017 Dec 28). Although endocrine therapy is the first-line treatment for estrogen receptor-positive breast cancer, about 30% of patients receiving adjuvant therapy experience recurrence, and almost all patients with metastatic breast cancer develop resistance and progress. The mechanisms of resistance to endocrine therapy can be divided into two categories. One category focuses on the estrogen receptor signaling pathway itself, including activation mutations, amplification, and fusion with other genes of the gene ESR1 encoding the estrogen receptor, as well as dysregulation of estrogen receptor co-regulators and downstream cell cycle control factors. The other category includes the activation of signaling pathways that cross-react with the estrogen receptor signaling pathway, such as the growth factor receptor pathway (Nat Rev Clin Oncol. 2015 Oct; 12(10):573-83).

[0005] Two studies in 2013 detected ESR1 gene mutations in 11-55% of estrogen receptor-positive metastatic breast cancer patients who had received aromatase inhibitor therapy. Further research revealed that the mutated receptor can phosphorylate independently of estrogen, exerting a transcriptional effect that allows estrogen-dependent MCF7-inoculated tumors to grow without relying on estrogen. Moreover, the mutated receptor reduces the activity of the SERM tamoxifen and SERD fulvestrant. Therefore, ESR1 gene mutation may be one of the mechanisms of drug resistance in estrogen-positive breast cancer (Nat Rev ClinOncol. 2015 Oct; 12(10):573-83 and Nat Genet 2013; 45:1439-45). In subsequent studies, a certain proportion of ESR1 gene mutations were found in estrogen receptor-positive metastatic breast cancer patients, with a mutation rate of approximately 30%. In the BOLERO-2 clinical trial, 29% of patients with estrogen receptor-positive metastatic breast cancer who progressed after AIs treatment had ER Y537S and ER D538G mutations in their ctDNA. In the exemestane monotherapy group, patients with mutations had shorter progression-free survival (PFS) and overall survival (OS) than patients without mutations [Nat Genet 2013; 45:1446-51].

[0006] In summary, ESR1 gene mutations mostly occur in patients with metastatic estrogen receptor-positive breast cancer who have progressed after AI therapy. These patients are no longer sensitive to AI therapy, so there is a need to develop estrogen receptor antagonists targeting ESR1 gene mutations.

[0007] Eisai's first-in-class estrogen receptor covalently binding antagonist H3B-6545 exhibits strong inhibitory activity against both wild-type and mutant estrogen receptors and can exert a longer-lasting effect through covalent binding to the receptor. It is currently undergoing phase I and II clinical trials. Currently, publicly disclosed patents for estrogen receptor antagonists targeting ESR1 gene mutations include WO2016196346 and WO2016196342.

[0008] PCT / CN2020 / 096744 provides an indazole derivative with the chemical name (E)-1-morpholino-4-((1-(((5-((Z)-4,4,4-trifluoro-1-(3-fluoro-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)amino)but-2-en-1-one (Formula I), offering patients a new treatment option.

[0009] Summary of the Invention

[0010] This disclosure provides a pharmaceutically acceptable salt of the compound represented by formula (I), said pharmaceutically acceptable salt being selected from maleate or hydrochloride.

[0011]

[0012] In an optional embodiment, this disclosure provides a maleate salt of the compound shown in formula (I), wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2 to 2:1.

[0013] In an optional embodiment, this disclosure provides a maleate salt of the compound shown in formula (I), wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:1.

[0014] This disclosure also provides a method for preparing the pharmaceutically acceptable salt of formula (I) described above, comprising: a step of forming a salt of the compound of formula (I) with an acid selected from maleic acid, hydrochloric acid or solutions thereof, and the solvent used in the salt-forming reaction selected from one or more of water, methanol, n-propanol, isopropanol, ethanol, isopropyl ether, tetrahydrofuran, isopropyl acetate, acetone, butanone, methyl tert-butyl ether, acetonitrile, 1,4-dioxane, ethyl acetate and n-hexane.

[0015] This disclosure further provides the I crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 16.448, 16.956, 19.332, 20.135, 21.645, 22.257 and 22.696.

[0016] Furthermore, the maleate salt of the compound shown in formula (I) is in crystal form I, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0017] This disclosure further provides the I crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.853, 16.448, 16.956, 19.332, 20.135, 20.835, 21.645, 22.257, 22.696 and 25.879.

[0018] Furthermore, the maleate salt of the compound shown in formula (I) is in crystal form I, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0019] This disclosure further provides the I crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.643, 7.853, 16.448, 16.956, 18.671, 19.332, 20.135, 20.835, 21.645, 22.257, 22.696, 25.879 and 29.015.

[0020] Furthermore, the maleate salt of the compound shown in formula (I) is in crystal form I, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0021] This disclosure further provides the II crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 15.005, 15.503, 17.599, 18.763, 20.471 and 26.259.

[0022] Furthermore, the maleate of the compound shown in formula (I) is in crystal form II, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0023] This disclosure further provides the II crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 8.724, 15.005, 15.503, 17.599, 18.136, 18.763, 20.471, 26.259 and 28.925.

[0024] Furthermore, the maleate of the compound shown in formula (I) is in crystal form II, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0025] This disclosure further provides the II crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 8.724, 15.005, 15.503, 17.599, 18.136, 18.763, 20.471, 22.600, 23.556, 24.643, 26.259 and 28.925.

[0026] Furthermore, the maleate of the compound shown in formula (I) is in crystal form II, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0027] This disclosure further provides the III crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 16.440, 17.358, 19.295, 19.919, 20.946 and 26.340.

[0028] Furthermore, the maleate of the compound shown in formula (I) is in crystal form III, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0029] This disclosure further provides the III crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 7.916, 16.440, 17.358, 19.295, 19.919, 20.946, 23.702, 25.820 and 26.340.

[0030] Furthermore, the maleate of the compound shown in formula (I) is in crystal form III, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0031] This disclosure further provides the III crystal form of the maleate of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 7.916, 14.150, 16.440, 17.358, 19.295, 19.919, 20.946, 22.943, 23.702, 25.820, 26.340 and 29.177.

[0032] Furthermore, the maleate of the compound shown in formula (I) is in crystal form III, wherein the molar ratio of the compound shown in formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0033] Another aspect of this disclosure provides a method for preparing the I crystal form of the maleate of the compound shown in formula (I), the method comprising the following steps:

[0034] 1) Dissolve the compound shown in formula (I) and maleic acid in a solvent selected from at least one of ethanol, acetone, methanol or water.

[0035] 2) Crystallization.

[0036] Furthermore, the maleate of the compound of formula (I) prepared above is in crystal form I, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0037] Another aspect of this disclosure provides a method for preparing the II crystal form of the maleate of the compound shown in formula (I), the method comprising the following steps:

[0038] 1) Dissolve the compound shown in formula (I) and maleic acid in a solvent selected from at least one of ethanol, isopropyl ether, n-hexane and butanone.

[0039] 2) Crystallization.

[0040] Furthermore, the maleate of the compound of formula (I) prepared above is in crystal form II, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0041] Another aspect of this disclosure provides a method for preparing the III crystal form of the maleate of the compound shown in formula (I), the method comprising heating the I crystal form of the maleate of the compound shown in formula (I) to 110°C.

[0042] Furthermore, the maleate of the compound of formula (I) prepared above is in crystal form III, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:2-2:1, preferably 1:1.

[0043] This disclosure provides a hydrochloride salt of the compound shown in formula (I), wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2 to 2:1.

[0044] In an optional embodiment, this disclosure provides a hydrochloride salt of the compound shown in formula (I), wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:1.

[0045] This disclosure further provides the α-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 12.578, 13.583, 18.151, 19.174, 20.027 and 26.978.

[0046] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form α, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0047] This disclosure further provides the α-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 8.647, 11.210, 12.578, 13.583, 18.151, 19.174, 20.027, 26.676 and 26.978.

[0048] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form α, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0049] This disclosure further provides the α-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 8.647, 11.210, 12.578, 13.583, 18.151, 19.174, 20.027, 24.105, 25.004, 25.375, 26.676 and 26.978.

[0050] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form α, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0051] This disclosure further provides the b-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186 and 20.074.

[0052] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form b, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0053] This disclosure further provides the b-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186, 20.074, 20.773, 21.186 and 26.977.

[0054] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form b, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0055] This disclosure further provides the b-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186, 20.074, 20.773, 21.186, 22.512, 24.181, 26.599 and 26.977.

[0056] Furthermore, the hydrochloride salt of the compound shown in formula (I) is in crystal form b, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0057] This disclosure further provides the c-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 15.515, 17.137, 19.743, 20.471, 21.525, 23.442 and 25.987.

[0058] This disclosure further provides the c-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.353, 13.066, 14.042, 15.515, 17.137, 19.743, 20.471, 21.525, 23.442 and 25.987.

[0059] This disclosure further provides the c-crystal form of the hydrochloride salt of the compound shown in formula (I), whose X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.970, 7.353, 9.527, 13.066, 14.042, 15.515, 17.137, 19.743, 20.471, 21.525, 23.442, 25.987 and 29.252.

[0060] Another aspect of this disclosure provides a method for preparing the α-crystal form of the hydrochloride salt of the compound shown in formula (I), the method comprising the following steps:

[0061] 1) Dissolve the compound of formula (I) in a solvent selected from at least one of ethanol, diethyl ether, butanone, n-hexane, or isopropanol.

[0062] 2) Add hydrochloric acid, and crystals will precipitate.

[0063] Furthermore, the hydrochloride salt of the compound shown in formula (I) prepared above is in crystal form α, wherein the molar ratio of the compound shown in formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0064] Another aspect of this disclosure provides a method for preparing the b-crystal form of the hydrochloride salt of the compound shown in formula (I), the method comprising the following steps:

[0065] 1) Dissolve the compound of formula (I) in a solvent selected from at least one of isopropanol, isopropyl ether, n-hexane, ethyl acetate, and acetonitrile.

[0066] 2) Add hydrochloric acid, and crystals will precipitate out.

[0067] Furthermore, the hydrochloride salt of the compound of formula (I) prepared above is in crystal form b, wherein the molar ratio of the compound of formula (I) to hydrochloric acid is 1:2-2:1, preferably 1:1.

[0068] Another aspect of this disclosure provides a method for preparing the c-crystal form of the hydrochloride salt of the compound shown in formula (I), the method comprising the following steps:

[0069] 1) Dissolve the compound shown in formula (I) in at least one solvent selected from acetone, methanol, and water.

[0070] 2) Add hydrochloric acid solution, and crystals will precipitate.

[0071] In an optional embodiment, the crystal form of a pharmaceutically acceptable salt of the compound represented by formula (I) is provided, wherein the error range of the 2θ angle is ±0.2.

[0072] In some embodiments, the method for preparing the crystal form described in this disclosure further includes filtration, washing, or drying steps.

[0073] This disclosure also provides pharmaceutical compositions prepared from pharmaceutically acceptable salts or crystal forms of the compounds represented by formula (I) above.

[0074] This disclosure also provides a pharmaceutical composition comprising: i) a pharmaceutically acceptable salt or a crystal form of a pharmaceutically acceptable salt of the compound represented by formula (I) above, and ii) optionally a pharmaceutically acceptable carrier, diluent or excipient.

[0075] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned component i) and component ii).

[0076] This disclosure also provides the use of a pharmaceutically acceptable salt of the compound represented by formula (I) or a crystal form of a pharmaceutically acceptable salt of the compound represented by formula (I), or the aforementioned composition, or a composition prepared by the aforementioned method, in the preparation of an estrogen receptor modulator.

[0077] This disclosure also provides the use of a pharmaceutically acceptable salt of the compound represented by formula (I) or a crystal form of a pharmaceutically acceptable salt of the compound represented by formula (I) or the aforementioned composition or a composition prepared by the aforementioned method in the preparation of a medicament for the prevention and / or treatment of estrogen receptor-mediated or dependent diseases or conditions, preferably said estrogen receptor-mediated or dependent diseases or conditions being cancer, more preferably breast cancer, ovarian cancer, endometrial cancer, prostate cancer or uterine cancer, and most preferably breast cancer.

[0078] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is ±0.20, and can be -0.20, -0.19, -0.18, -0.17, -0.16, -0.15, -0.14, -0.13, -0.12, -0.11, -0.10, -0.09, -0.08, -0.07, -0.06, -0.05, -0.04, -0.03, -0.02, -0.01, 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20.

[0079] The “crystallization” described in this disclosure includes, but is not limited to, agitation crystallization, pulping crystallization, and evaporation crystallization.

[0080] The method for preparing the crystal form described in this disclosure also includes steps such as filtration and drying.

[0081] The drying temperature described in this disclosure is generally 25℃-100℃, preferably 40℃-70℃, and can be dried under normal pressure or reduced pressure. Attached Figure Description

[0082] Figure 1 XRPD spectrum of maleate of the compound shown in formula (I) in crystal form I;

[0083] Figure 2 XRPD spectrum of maleate of the compound shown in formula (I) in crystal form II;

[0084] Figure 3 XRPD spectrum of maleate of the compound shown in formula (I) in crystal form III;

[0085] Figure 4 XRPD spectra of the α-crystal form of the hydrochloride salt of the compound shown in formula (I);

[0086] Figure 5XRPD spectra of the b-crystal form of the hydrochloride salt of the compound shown in formula (I);

[0087] Figure 6 XRPD spectra of the c-crystal form of the hydrochloride salt of the compound shown in formula (I). Detailed Implementation

[0088] The present invention will be explained in more detail below with reference to the embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the nature and scope of the present invention.

[0089] Test conditions of the instruments used in the experiment:

[0090] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰. -6 The unit (ppm) is given. NMR determination was performed using a Bruker AVANCE-400 NMR spectrometer or a Bruker AVANCE NEO 500M. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0091] MS measurements were performed using an Agilent 1200 / 1290 DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometry system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS).

[0092] waters ACQuity UPLC-QD / SQD (Manufacturer: waters, MS model: waters ACQuity QdaDetector / waters SQ Detector)

[0093] THERMO Ultimate 3000-Q Exactive (Manufacturer: THERMO, MS Model: THERMO QExactive)

[0094] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLC e2695-2489 high-performance liquid chromatograph.

[0095] Chiral HPLC analysis was performed using an Agilent 1260 DAD high-performance liquid chromatograph.

[0096] High performance liquid chromatography (HPLC) was performed using Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson GX-281 preparative chromatographs.

[0097] Chiral preparation was performed using a Shimadzu LC-20AP preparative chromatograph.

[0098] Ion chromatography was performed using Thermo Scientific Dionex Intergrion, column model: DionexIonPacTM AS11-HC (4μm, 4×250cm).

[0099] XRPD (X-ray Powder Diffraction) was used for analysis. Measurements were performed using a BRUKER D8 Discover X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα rays. Scanning mode: θ / 2θ, scanning range (2θ range): 3~50°.

[0100] DSC stands for Differential Scanning Calorimetry: Measurements were performed using a METTLER TOLEDO DSC 3+ differential scanning calorimeter with a heating rate of 10℃ / min. The specific temperature range was referenced from the corresponding spectra (mostly 25-300 or 25-350℃). Nitrogen purging rate was 50mL / min.

[0101] TGA is thermogravimetric analysis: the test was performed using a METTLER TOLEDO TGA 2 thermogravimetric analyzer, with a heating rate of 10℃ / min, and the specific temperature range was referenced from the corresponding spectrum (mostly 25-300℃). The nitrogen purging rate was 50mL / min.

[0102] DVS stands for Dynamic Moisture Adsorption: Detection is performed using SMS DVS Advantage at 25℃, with humidity changes of 50%-95%-0%-95%-50%, in 10% increments (the final step is 5%) (the specific humidity range is subject to the corresponding spectrum; the methods listed here are the most commonly used). The judgment criterion is that dm / dt is not greater than 0.002%.

[0103] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0104] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0105] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compounds, and the developing solvent system for TLC included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system. The volume ratio of the solvent was adjusted according to the polarity of the compounds, and small amounts of basic or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0106] Example 1: Preparation of compound (E)-1-morpholinyl-4-((1-(((5-((Z)-4,4,4-trifluoro-1-(3-fluoro-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)amino)but-2-en-1-one

[0107]

[0108] Step 1: (1-(((5-iodopyridin-2-yl)oxy)methyl)cyclopropyl)tert-butyl carbamate 1c

[0109] Sodium hydride (0.4 g, 10.7 mmol) was dissolved in N,N-dimethylformamide (20 mL), and tert-butyl 1-(hydroxymethyl)cyclopropylcarbamate 1b (1.0 g, 5.3 mmol, prepared by a known method, "Journal of Organic Chemistry, 2002, 67(11), 3965-3968") was added at room temperature. After the addition was complete, 2-fluoro-5-iodopyridine 1a (1.8 g, 8.0 mmol) was slowly added. The reaction was stopped after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer chromatography using solvent system B to obtain the title product 1c (2.4 g), yield: 86%.

[0110] MS m / z(ESI): 391.0 [M+1]

[0111] Step 2: (Z)-(1-(((5-(4,4,4-trifluoro-1-(3-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)tert-butyl carbamate 1f

[0112] 3-Fluoro-1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,4-trifluorobut-1-yn-1-yl)-1H-indazole 1d (1.8 g, 5.5 mmol, prepared by the method disclosed in Example 3 on page 84 of patent application WO2018098305) was dissolved in methyltetrahydrofuran (40 mL), and bis-pinacol boronic acid ester (1.7 g, 6.6 mmol) and tetra-triphenylphosphine platinum (137 mg, 0.1 mmol) were added. The mixture was purged with argon three times, heated to 85 °C, and stirred for 3 hours. After cooling to room temperature, compound 1c (2.0 g, 5.2 mmol), bis-triphenylphosphine palladium dichloride (741 mg, 1.1 mmol), cesium carbonate (3.6 g, 11.0 mmol), and water (1 mL) were added, and the mixture was stirred overnight at room temperature. Iodobenzene 1e (1.2 g, 6.1 mmol) and potassium hydroxide (1.5 g, 27.6 mmol) were added. The mixture was purged with argon three times, heated to 85 °C, stirred for 2 hours, and then cooled to room temperature to stop the reaction. The reaction solution was concentrated under reduced pressure, and the residue was purified by thin-layer chromatography using solvent system B to obtain the title product 1f (3.0 g), yield: 88%.

[0113] MS m / z (ESI): 667.2 [M+1]

[0114] Step 3: (Z)-(1-(((5-(4,4,4-trifluoro-1-(3-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)-1-amine 1g

[0115] Compound 1f (1.8 g, 2.7 mmol) was dissolved in dichloromethane (15 mL), and trifluoroacetic acid (3 mL) was added. The mixture was stirred at room temperature for 5 hours, and then the reaction was stopped. The reaction solution was concentrated under reduced pressure, and the pH was adjusted to approximately 8 with saturated sodium bicarbonate solution (100 mL). The solution was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 1 g of crude product (1.4 g), yield: 89%. The product was used directly in the next reaction without further purification.

[0116] Step 4: (E)-1-morpholino-4-((1-(((5-((Z)-4,4,4-trifluoro-1-(3-fluoro-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)amino)but-2-en-1-one 1i

[0117] 1 g (1.7 g, 2.8 mmol) of compound was dissolved in N,N-dimethylformamide (20 mL), and diisopropylethylamine (1.1 g, 8.5 mmol) was added at room temperature. Then (E)-4-bromo-1-morpholinylbut-2-en-1-one (0.7 g, 2.8 mmol, prepared by the method disclosed in Example 15 on page 65 of patent application US2016347717) was added, and the reaction was stirred for 2 hours. The reaction was stopped and cooled, and saturated sodium bicarbonate solution (15 mL) was added. The mixture was extracted with ethyl acetate (50 mL × 2), the organic phases were combined, washed with saturated sodium chloride solution (50 mL × 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin-layer chromatography using solvent system A to give the title product 1i (1.3 g), yield: 65%.

[0118] MS m / z(ESI): 720.2 [M+1]

[0119] Step 5: (E)-1-morpholino-4-((1-(((5-((Z)-4,4,4-trifluoro-1-(3-fluoro-1H-indazol-5-yl)-2-phenylbut-1-en-1-yl)pyridin-2-yl)oxy)methyl)cyclopropyl)amino)but-2-en-1-one I

[0120] Compound 1i (2.0 g, 2.8 mmol) was dissolved in methanol (5 mL), and hydrochloric acid (12 N, 10 mL) was added. The mixture was stirred for 3 hours. The reaction was stopped and cooled. The reaction solution was concentrated, and saturated sodium bicarbonate solution (15 mL) was added. The mixture was extracted with dichloromethane (50 mL × 4). The organic phases were combined and washed successively with water (30 mL × 3), saturated sodium chloride solution (50 mL), and dried over anhydrous sodium sulfate. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by thin-layer chromatography using solvent system A to give title product 1 (1.3 g), yield: 73%.

[0121] MS m / z(ESI): 636.2 [M+1];

[0122] 1 ¹H NMR (400MHz, CD₃OD) 7.65 (d, 2H), 7.49 (d, 1H), 7.30–7.22 (m, 7H), 6.82–6.76 (m, 1H), 6.60–6.52 (m, 2H), 4.15 (s, 2H), 3.62–3.39 (m, 12H), 0.76–0.64 (m, 4H). X-ray powder diffraction analysis showed that the product was amorphous.

[0123] Test Example 1: Determination of the inhibitory effect of compound I on estrogen receptor reporter gene activity

[0124] 1. Experimental Objective

[0125] The purpose of this experiment is to test the inhibitory effect of the disclosed compound on the activity of the estrogen receptor reporter gene, based on IC50. 50 Size is used to evaluate the in vitro activity of compounds.

[0126] 2. Experimental Methods

[0127] MCF7 cells (ATCC, HTB-22) expressing the estrogen receptor response element-controlled luciferase reporter gene ERE-luc (synthesized by Genewiz Biotechnology Co., Ltd.) were cultured in MEM (GE Healthcare, SH30024.01) medium containing 10% fetal bovine serum and 500 μg / ml G418. On the first day of the experiment, MCF7 / ERE-luc cells were seeded at a density of 30,000 cells / well in 96-well plates using incomplete MEM medium containing 10% activated charcoal-treated fetal bovine serum (BioSun, BS-0004-500), with 100 μl of cell suspension per well, and incubated overnight at 37°C in a 5% CO2 cell culture incubator. On the second day, 10 μl of β-estradiol prepared in incomplete culture medium and different concentrations of the test compound were added to each well. The final concentration of β-estradiol was 0.1 nM, and the final concentrations of the compounds were nine concentration points obtained by serially diluting the compounds 10-fold from 10 μM. A blank control containing 0.5% DMSO was included. The cells were incubated at 37°C and 5% CO2 for 20 hours. On the third day, the 96-well plate was removed, and 100 μl of ONE-Glo was added to each well. TM The activity of luciferase was detected using the Luciferase Assay System (Promega, E6110). After cells were fully lysed at room temperature for 3 minutes, the luminescence signal values ​​were read using a multi-label microplate reader (PerkinElmer, VICTOR 3). The IC50 of the compound's inhibitory activity was calculated using Graphpad Prism software based on the compound concentration and the luminescence signal value. 50 value.

[0128] 3. Test Results

[0129] The inhibitory effect of the compounds in this disclosure on the activity of estrogen receptor reporter genes was determined through the above experiments. The chemiluminescence signal value was plotted against the logarithmic concentration of the compounds using a Graghpad Prism analyzer, and the IC50 of compound I was measured. 50 The value is 1nM.

[0130] Therefore, the disclosed compound has a significant inhibitory effect on the estrogen receptor reporter gene.

[0131] Test Example 2: Inhibitory effect of the disclosed compound on MCF7 cell proliferation

[0132] 1. Experimental Objective

[0133] The purpose of this experiment was to determine the inhibitory activity of the disclosed compound on the proliferation of MCF7 cells, based on IC50. 50 Size is used to evaluate the in vitro activity of compounds.

[0134] 2. Experimental Methods

[0135] MCF7 cells (ATCC, HTB-22) were cultured in MEM (GE Healthcare, SH30024.01) complete medium containing 10% fetal bovine serum. On day 1, MCF7 cells were seeded at a density of 3,000 cells / well in 96-well plates using complete medium, with 100 μl of cell suspension per well. The plates were incubated overnight at 37°C with 5% CO2. On day 2, the medium was removed, and each well was replaced with 135 μl of MEM incomplete medium containing 2% fetal bovine serum. Simultaneously, 15 μl of different concentrations of the test compound prepared in incomplete medium was added to each well. The final concentrations of the compounds were determined by a 4-fold serial dilution starting from 100 nM (9 concentration points). A blank control containing 0.5% DMSO was included. The plates were incubated at 37°C with 5% CO2 for 144 hours. On day 8, the 96-well cell culture plates were removed, and 150 μl of the test compound was added to each well. The Luminescent CellViability Assay (Promega, G7573) was incubated at room temperature for 10 minutes. The luminescence signal values ​​were then read using a multi-label microplate reader (PerkinElmer, VICTOR 3). The IC50 of the compound's inhibitory activity was calculated using Graphpad Prism software based on the compound's concentration and luminescence signal value. 50 value.

[0136] 3. Data Analysis

[0137] The chemiluminescence signal value was plotted against the logarithmic concentration of the compound using Graghpad Prism to obtain the IC50 of the compound. 50 The value was 0.5 nM, indicating that the compound disclosed herein has a significant inhibitory effect on the proliferation of MCF7 cells.

[0138] Test Example 3: Biological Evaluation of the Proliferation Inhibition Experiment of MCF7 Cells Expressing ERα Mutant

[0139] 1. Experimental Objective

[0140] The purpose of this experiment is to determine the inhibitory activity of the disclosed compound on the proliferation of ERα mutant MCF7 cells.

[0141] 2. Experimental Methods

[0142] Site-directed mutagenesis and cell line construction

[0143] The mutants ERαY537S and ERαD538G of the human estrogen receptor α (ERα) protein were obtained by site-directed mutagenesis using two-primer PCR with wild-type ESR1 gene cDNA (Accession No. NM000125) as a template. The primer sequences used for mutation are as follows (underlined nucleotides indicate the mutation sites): Y537S: F-AAGAAC GTG GTG CCC CTC T C T GAC CTG CTG CTG GAG ATG; R-CAT CTC CAG CAG CAG GTC A G AGAG GGG CAC CAC GTT CTT; D538G:F-AAC GTG GTG CCC CTC TAT G G C CTG CTG CTG GAGATG CTG; R-CAG CAT CTC CAG CAG CAG G C C ATA GAG GGG CAC CAC GTT. The cDNA of the mutant ESR1 was cloned into the target lentiviral vector pCDH-CMV-MCS-EF1-Puro. Then, the lentiviral plasmid containing the mutant ESR1 gene sequence and the lentiviral packaging plasmid were transfected into HEK-293T cells (ATCC, CRL-3216) using Lipofectamine 3000 Transfection Reagent (ThermoFisher Scientific, Cat# L3000075). Forty-eight hours post-transfection, the virus-containing culture supernatant was filtered, ultracentrifuged to obtain the viral pellet, resuspended in an appropriate amount of culture medium, and added to MCF7 cells (ATCC, HTB-22). Polybrene was added to a final concentration of 8 μg / ml and incubated overnight. Two days after transfection, 1 μg / ml of puromycin was added to the cell culture medium for resistance selection. About two weeks later, MCF7 cell lines that could stably express ERαY537S and ERαD538G mutants were obtained.

[0144] Cell proliferation inhibition experiment

[0145] MCF7 cells expressing the ERα mutant were cultured in MEM (GE Healthcare, SH30024.01) complete medium containing 10% fetal bovine serum. On day 1, cells were seeded at a density of 3,000 cells / well in 96-well plates using complete medium, with 100 μl of cell suspension per well, and incubated overnight at 37°C with 5% CO2. On day 2, the medium was aspirated, and each well was replaced with 135 μl of MEM incomplete medium containing 2% fetal bovine serum. Simultaneously, 15 μl of different concentrations of the test compound prepared in incomplete medium was added to each well. The final concentrations of the compounds were determined by a 4-fold serial dilution starting from 100 nM, with a blank control containing 0.5% DMSO. The plates were incubated at 37°C with 5% CO2 for 144 hours. On day 8, the 96-well cell culture plates were removed, and 150 μl of the test compound was added to each well. The Luminescent CellViability Assay (Promega, G7573) was incubated at room temperature for 10 minutes. The luminescence signal values ​​were then read using a multi-label microplate reader (PerkinElmer, VICTOR 3). The IC50 of the compound's inhibitory activity was calculated using Graphpad Prism software based on the compound's concentration and luminescence signal value. 50 The IC50 value of the disclosed compound on the inhibitory effect of its compound on the proliferation of MCF7 D538G cells expressing the ERα mutant is [value missing]. 50 The IC50 value was 2 nM, representing the inhibitory effect on the proliferation of MCF7ERαY537S cells expressing the ERα mutant. 50 The concentration was 3 nM, and the results showed that the compound disclosed herein had a significant inhibitory effect on the proliferation of MCF7 cells expressing the ERα mutant.

[0146] Test Example 4: Pharmacokinetic assay of the disclosed compound in BALB / c nude mice

[0147] 1. Abstract

[0148] Using BALB / c nude mice as test animals, the plasma drug concentration of Compound I at different time points after gavage administration was determined by LC / MS / MS. The pharmacokinetic behavior of Compound I in BALB / c nude mice was investigated to evaluate its pharmacokinetic characteristics.

[0149] 2. Test Plan

[0150] 2.1 Test Drugs

[0151] Compound of formula I.

[0152] 2.2 Experimental Animals

[0153] 36 female BALB / C nude mice were evenly divided into 4 groups with 9 mice in each group. They were purchased from Jieshijie Laboratory Animal Co., Ltd., and the animal production license number is SCXK(Shanghai)2013 - 0006.

[0154] 2.3 Drug preparation

[0155] Weigh an appropriate amount of the sample and dissolve it in 5% volume of DMSO, 5% volume of Tween 80, and 90% volume of normal saline to prepare a colorless, clear and transparent liquid with a concentration of 0.1 mg / mL.

[0156] 2.4 Drug administration

[0157] After fasting overnight, intragastric administration was carried out respectively. The administration volume was 0.2 ml / 10 g, and the administration dose of the compound of formula I was 30 mg / kg.

[0158] 3. Operation

[0159] 36 female Balb / C nude mice; after fasting overnight, intragastric administration was carried out. Blood samples of 0.1 ml were collected at 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, 24.0 h after administration (3 animals at each time point), placed in heparinized test tubes, centrifuged at 3500 rpm for 10 min to separate plasma, and stored at -20 °C. To determine the content of the test compound in the plasma of nude mice after intragastric administration of different concentrations of the drug: Take 25 μL of the plasma of nude mice at each time point after administration, add 40 μL of the internal standard solution camptothecin (100 ng / mL), 200 μL of acetonitrile, vortex mix for 5 minutes, centrifuge for 10 minutes (4000 revolutions per minute), and take 0.5 μL of the supernatant of the plasma sample for LC / MS / MS analysis.

[0160] 4. Results of pharmacokinetic parameters of BALB / C nude mice

[0161] The pharmacokinetic parameters of the compound of formula I disclosed in the present invention are as follows:

[0162]

[0163] Conclusion: The compound disclosed in the present invention has good pharmacokinetic absorption and obvious pharmacokinetic absorption effect.

[0164] Test Example 5: Biological evaluation of covalent modification of estrogen receptor ERα wild type and ERαY537S mutant

[0165] 1. Experimental purpose

[0166] The purpose of this experiment is to determine the covalent modification effect of the compound disclosed in the present invention on estrogen receptor ERα wild type and ERαY537S mutant.

[0167] 2. Experimental method

[0168] The ligand-binding domain (LBD, aa296-554) of the estrogen receptor ERα wild-type and ERαY537S mutant was expressed and purified by *E. coli*. 2 μM of wild-type ERα or ERαY537S mutant protein and 10 μM of the compound were added to a buffer solution containing 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM TCEP, and 5% glycerol, mixed, and incubated at 4°C for 24 hours before high-resolution mass spectrometry (HRMS) detection. Alternatively, 1 μM of wild-type ERα or ERαY537S mutant protein and 3 μM of the compound were added to a buffer solution containing 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1 mM TCEP, and 5% glycerol, mixed, and incubated at 37°C for 15 minutes before HMS detection. In the mass spectrometry results, the peak with a molecular weight equal to the sum of the protein and the compound is the covalently modified product. The percentage of covalent modification can be calculated by calculating the ratio of unbound protein to total protein.

[0169] Covalent modification ratio 24 hours after covalent modification:

[0170]

[0171] Conclusion: The tested compounds exhibited good covalent modification effects on both wild-type and mutant ERα proteins.

[0172] Example 2: Preparation of Maleate I crystal form

[0173] Weigh 10 mg of compound (I) and 3.0 mg of maleic acid ligand, add 500 μL of ethanol solvent and stir until dissolved. Stir for 2 days, allowing slow evaporation, and gradually precipitate the solid. Filter and vacuum dry the solid to obtain the product. 1 H-NMR showed that the molar ratio of the compound to maleic acid was 1:1.

[0174] X-ray powder diffraction analysis determined the product to be crystal form I. The XRPD spectrum is shown below. Figure 1 The peak positions are shown in Table 1.

[0175] Table 1. Peak positions of maleate I crystal form

[0176]

[0177]

[0178] The DSC spectrum shows endothermic peaks at 61.47℃ and 115.48℃; the TGA spectrum shows a weight loss of 2.81% between 25℃ and 140℃.

[0179] Example 3: Preparation of Maleate I crystal form

[0180] Weigh 10 mg of compound (I) and 3.0 mg of maleic acid ligand, add 500 μL of 10% water / acetone solvent and stir until dissolved. Stir for 2 days, allowing slow evaporation, and gradually precipitate the solid. Filter and vacuum dry the solid. NMR data show that the molar ratio of compound (I) to maleic acid in this salt is 1:1. X-ray powder diffraction analysis confirms it is crystal form I.

[0181] Example 4: Preparation of Maleate I crystal form

[0182] Weigh 10 mg of compound (I) and 3.0 mg of maleic acid ligand, add 500 μL of 10% water / methanol solvent and stir until dissolved. Stir for 2 days, allowing slow evaporation, and gradually precipitate the solid. Filter and vacuum dry the solid. NMR data show that the molar ratio of compound (I) to maleic acid in this salt is 1:1. X-ray powder diffraction analysis confirms it is crystal form I.

[0183] Example 5: Preparation of Maleate II Crystal Form

[0184] The compound shown in Formula I (25 mg, 39.33 μmol) was added to a mixed solvent of 1.5 mL of ethanol and n-hexane (V / V = 1:1), stirred, and 0.5 mL of maleic acid (4.57 mg, 39.33 μmol) butanone solution was added dropwise. The mixture was stirred until dissolved, and 0.5 mL of n-hexane was added and stirred. After 0.5 hours, a white turbid liquid gradually formed. The mixture was stirred at room temperature for 16 hours, filtered, and the filter cake was collected and dried under vacuum to obtain the title product (25 mg, yield: 100%).

[0185] The product obtained 1 H-NMR characterization and NMR data indicate that the molar ratio of the main component to maleic acid in this salt is 1:1.

[0186] 1 H NMR (400MHz, CD3OD) δ7.73-7.64(m,2H),7.52(dd,1H),7.32(dd,2H),7.26(d,4H),7.22(dt,1H),6.82(d,1H),6.75-6.63( m,2H),6.27(s,2H),4.41(s,2H),4.01(d,2H),3.69-3.61(m,8H),3.50-3.35(m,2H),1.19-1.17(m,2H),1.16-1.05(m,2H).

[0187] X-ray powder diffraction analysis determined that the product was crystal form II, such as... Figure 2 The peak positions are shown in Table 2.

[0188] The DSC spectrum shows an endothermic peak at 136.02℃; the TGA spectrum shows a weight loss of 0.58% at 25℃-85℃, 3.73% at 85℃-150℃, and 4.39% at 150℃-210℃.

[0189] Table 2. Peak positions of maleate II crystal form

[0190]

[0191]

[0192] Example 6: Preparation of Maleate III Crystal Form

[0193] The crystal form of compound I, as shown in Formula I, was heated to 110°C using DSC. X-ray powder diffraction analysis revealed a crystal form transformation, which was defined as crystal form III. Figure 3 The peak positions are shown in Table 3. The obtained products... 1 H-NMR characterization and NMR data indicate that the molar ratio of the main component to maleic acid in this salt is 1:1.

[0194] Table 3. Peak positions of maleate III crystal form

[0195]

[0196] Example 7: Preparation of the a-crystal form of hydrochloride

[0197] The compound shown in Formula I (25 mg, 39.33 μmol) was added to 2 mL of a mixed solvent of ethanol and diethyl ether (V / V = 1:1), stirred, and dissolved. Then, 3.28 μL of concentrated hydrochloric acid (12 M, 39.33 mmol, 35%) was slowly added dropwise, resulting in a white turbid liquid that gradually became viscous. Stirring continued, and after 0.5 hours, a white turbid liquid gradually formed. The mixture was stirred at room temperature for 16 hours, filtered, and the filter cake was collected and dried under vacuum to obtain the title product (23 mg, yield: 80%). X-ray powder diffraction analysis identified the product as crystal form a, as shown in the image. Figure 4 The peak positions are shown in Table 4.

[0198] Table 4. Peak positions of hydrochloride a crystal form

[0199]

[0200]

[0201] Example 8: Preparation of the a-crystal form of hydrochloride

[0202] The compound shown in Formula I (1.5 g, 2.36 mmol) was added to 10 mL of butanone, stirred, heated to 60 °C, and dissolved. Then, 0.5 mL of n-hexane was added, cooled to 40 °C, and 229.36 μL of concentrated hydrochloric acid (270.41 mg, 2.60 mmol, 35%) was slowly added dropwise until dissolved. The mixture was then cooled to room temperature and stirred at room temperature for 24 hours. No solid precipitated. The hydrochloride seed crystals of the compound shown in Formula I (Example 11) were added, and a white solid gradually precipitated. The mixture was stirred for 16 hours, and a white turbid liquid gradually formed. The mixture was filtered, the filter cake was collected, and vacuum dried to obtain the title product (1 g, yield: 63%).

[0203] Ion chromatography analysis showed that the chloride ion content was 4.99%, indicating that the molar ratio of the main component to hydrochloric acid in the salt was 1:1.

[0204] X-ray powder diffraction analysis revealed that the product was crystal form a.

[0205] The DSC spectrum showed an endothermic peak at 173.48℃; the TGA spectrum showed a weight loss of 2.81% between 25℃ and 170℃, and 2.07% between 170℃ and 200℃. DVS analysis showed that under normal storage conditions (25℃, 60% RH), the sample gained approximately 0.89% of its weight due to moisture absorption; under accelerated testing conditions (70% RH), the weight gain was approximately 1.02%; and under extreme conditions (90% RH), the weight gain was approximately 1.49%. Re-testing of the crystal form after DVS analysis showed no change in crystal form.

[0206] Example 9: Preparation of the a-crystal form of hydrochloride

[0207] The compound shown in Formula I (25 mg, 39.33 μmol) was added to 2 mL of isopropanol and stirred until dissolved. Then, 3.28 μL of concentrated hydrochloric acid (12 M, 39.33 mmol, 35%) was added dropwise, resulting in a white turbid liquid that gradually became viscous. Stirring continued, and after 0.5 hours, a white turbid liquid gradually formed. The mixture was stirred at room temperature for 16 hours, filtered, and the filter cake was collected and dried under vacuum to obtain the title product (23 mg, yield: 80%). X-ray powder diffraction analysis showed that the product was of crystal form a.

[0208] Example 10: Preparation of the a-crystal form of hydrochloride

[0209] The compound shown in Formula I (25 mg, 39.33 μmol) was added to 2 mL of butanone and stirred until dissolved. Then, 3.28 μL of concentrated hydrochloric acid (12 M, 39.33 mmol, 35%) was added dropwise, resulting in a white turbid liquid that gradually became viscous. Stirring continued, and after 0.5 hours, a white turbid liquid gradually formed. The mixture was stirred at room temperature for 16 hours, filtered, and the filter cake was collected and dried under vacuum to obtain the title product (23 mg, yield: 80%). X-ray powder diffraction analysis showed that the product was crystal form a.

[0210] Example 11: Preparation of the b-type hydrochloride

[0211] The compound shown in Formula I (1.2 g, 1.89 mmol) was added to a mixed solvent of isopropanol, isopropyl ether, and n-hexane (V / V / V = 1:1:1), stirred, and dissolved. Then, 173.05 μL of concentrated hydrochloric acid (12 M, 2.07 mmol, 35%) was added dropwise, and the solution was dissolved. The mixture was stirred at room temperature for 24 hours, and a white turbid liquid gradually formed. The solution was filtered, the filter cake was collected, and dried under vacuum to give the title product (1.1 g, yield: 86.69%).

[0212] Ion chromatography analysis showed that the chloride ion content was 5.04%, indicating that the molar ratio of the main component to hydrochloric acid in the salt was 1:1.

[0213] X-ray powder diffraction analysis determined the product to be crystal form b. The XRPD spectrum is shown below. Figure 5 The peak positions are shown in Table 5.

[0214] The DSC spectrum showed that the endothermic peak was not obvious, and the exothermic peak had a peak value of 183.54℃; the TGA spectrum showed a weight loss of 0.58% from 25℃ to 145℃ and a weight loss of 1.10% from 145℃ to 180℃.

[0215] DVS testing showed that under normal storage conditions (25°C, 60% RH), the sample's moisture absorption weight gain was approximately 0.84%; under accelerated testing conditions (70% RH), the moisture absorption weight gain was approximately 0.99%; and under extreme conditions (90% RH), the moisture absorption weight gain was approximately 1.8%. During humidity variations from 0% to 95% RH, the desorption and adsorption processes of the sample essentially overlapped. Re-testing of the crystal form after DVS testing showed no change in crystal form.

[0216] Table 5. Peak positions of b-type hydrochloride crystals

[0217]

[0218]

[0219] Example 12: Preparation of the b-type hydrochloride

[0220] The compound shown in Formula I (50 mg, 78.66 μmol) was added to a mixed solvent of n-hexane and ethyl acetate (V / V = 1:1), stirred until dissolved, and then 6.55 μL of concentrated hydrochloric acid (12 M, 78.66 mmol, 35%) was slowly added dropwise. A white turbid liquid appeared, which gradually became viscous. After stirring for 0.5 hours, a white turbid liquid gradually formed. The mixture was stirred at room temperature for 16 hours, filtered, and the filter cake was collected and dried under vacuum to obtain the title product (45 mg, yield: 85.12%). X-ray powder diffraction analysis showed that the product was crystal form b.

[0221] Example 13: Preparation of the b-type hydrochloride

[0222] The compound shown in Formula I (50 mg, 78.66 μmol) was added to 2 mL of acetonitrile, stirred, and dissolved completely. Then, 7.21 μL of concentrated hydrochloric acid (12 M, 86.5 mmol, 35%) was slowly added dropwise, and the solution remained dissolved. The mixture was stirred at room temperature for 24 hours, after which a white turbid liquid gradually formed. The solution was filtered, the filter cake was collected, and dried under vacuum to obtain the title product (40 mg, yield: 75.66%). X-ray powder diffraction analysis showed that the product was crystal form b.

[0223] Example 14: Preparation of the c-crystal form of hydrochloride

[0224] Weigh 15 mg of the compound shown in Formula I, add 500 μl of solvent, stir until clear, add 26 μl of 1M hydrochloric acid aqueous solution (no precipitation), and slowly evaporate to obtain the product. X-ray powder diffraction analysis confirms the product as crystal form c. The XRPD spectrum is shown below. Figure 6 The peak positions are shown in Table 6.

[0225] The DSC spectrum shows endothermic peaks at 94.07℃ and 147.06℃; the TGA spectrum shows a weight loss of 5.30% between 25℃ and 120℃.

[0226] DVS testing showed that under normal storage conditions (25°C, 60% RH), the sample's moisture absorption weight gain was approximately 8.64%; under accelerated testing conditions (70% RH), the moisture absorption weight gain was approximately 9.32%; and under extreme conditions (90% RH), the moisture absorption weight gain was approximately 12.32%. During the humidity variation from 0% to 95% RH, the desorption and adsorption processes of the sample essentially overlapped. Re-testing of the crystal form after DVS testing showed no change in crystal form.

[0227] Table 6. Peak positions of C-type hydrochloride crystals

[0228]

[0229] Example 15: Experiment on Factors Affecting the Crystal Form of Hydrochloride a

[0230] Take the hydrochloride crystal form a of compound of formula 1 into an open, clean weighing bottle and investigate the stability of the sample under high temperature (40℃, 60℃), light (4500lx±500lx), and high humidity (90%±5%RH, 75%±5%RH) conditions. The sampling period is 30 days, and the results are shown in Table 7 below.

[0231] Table 7. Results of 30-day test on factors influencing the crystal form a of compound I hydrochloride.

[0232]

[0233] Experimental results show that the hydrochloride crystal form a of compound I exhibits good physical and chemical stability under light, 40℃, 60℃, 75%RH, and 90%RH conditions.

[0234] Example 16: Long-term accelerated stability test of hydrochloride crystal form a

[0235] The hydrochloride crystal form a of the compound shown in Formula I was subjected to a 6-month long-term (25℃, 60%RH) and accelerated (40℃, 75%RH) stability study, and the results are shown in Table 8 below.

[0236] Table 8

[0237]

[0238] The results showed that the hydrochloride crystal form a sample exhibited good physicochemical stability after being placed under long-term (25℃, 60%RH) and accelerated (40℃, 75%RH) conditions for 6 months.

Claims

1. A pharmaceutically acceptable salt of a compound of formula (I), said pharmaceutically acceptable salt being selected from maleate and hydrochloride salts.

2. A pharmaceutically acceptable salt of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:2 to 2:

1.

3. A pharmaceutically acceptable salt of the compound of formula (I) according to claim 2, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:

1.

4. A maleate salt of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:1, and its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 16.448, 16.956, 19.332, 20.135, 21.645, 22.257 and 22.

696.

5. The maleate of the compound of formula (I) according to claim 4, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.853, 16.448, 16.956, 19.332, 20.135, 20.835, 21.645, 22.257, 22.696 and 25.

879.

6. The maleate of the compound of formula (I) according to claim 4, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.643, 7.853, 16.448, 16.956, 18.671, 19.332, 20.135, 20.835, 21.645, 22.257, 22.696, 25.879 and 29.

015.

7. A maleate crystal form II of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:1, and its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 15.005, 15.503, 17.599, 18.763, 20.471 and 26.

259.

8. The maleate of the compound of formula (I) according to claim 7, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 8.724, 15.005, 15.503, 17.599, 18.136, 18.763, 20.471, 26.259 and 28.

925.

9. The maleate of the compound of formula (I) according to claim 7, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.440, 8.724, 15.005, 15.503, 17.599, 18.136, 18.763, 20.471, 22.600, 23.556, 24.643, 26.259 and 28.

925.

10. A maleate crystal form III of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to maleic acid is 1:1, and its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 16.440, 17.358, 19.295, 19.919, 20.946 and 26.

340.

11. The maleate of the compound of formula (I) according to claim 10, in crystal form III, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 7.916, 16.440, 17.358, 19.295, 19.919, 20.946, 23.702, 25.820 and 26.

340.

12. The maleate of the compound of formula (I) according to claim 10, in crystal form III, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.830, 7.916, 14.150, 16.440, 17.358, 19.295, 19.919, 20.946, 22.943, 23.702, 25.820, 26.340 and 29.

177.

13. A method for preparing the I crystal form of the maleate of the compound of formula (I) according to any one of claims 4-6, the method comprising the following steps: 1) Dissolve the compound shown in formula (I) and maleic acid in a solvent selected from at least one of ethanol, acetone, methanol or water. 2) Crystallization.

14. A method for preparing the II crystal form of the maleate of the compound of formula (I) according to claims 7-9, the method comprising the following steps: 1) Dissolve the compound shown in formula (I) and maleic acid in a mixed solvent selected from ethanol and n-hexane. 2) Crystallization.

15. A method for preparing the III crystal form of the maleate of the compound of formula (I) according to any one of claims 10-12, the method comprising the step of heating the I crystal form of the maleate of the compound of formula (I) to 110°C.

16. A pharmaceutically acceptable salt of the compound of formula (I) according to claim 1, wherein the hydrochloride salt is wherein the molar ratio of the compound of formula (I) to hydrochloric acid is 1:2 to 2:

1.

17. A pharmaceutically acceptable salt of the compound of formula (I) according to claim 16, wherein the molar ratio of the compound of formula (I) to hydrochloric acid is 1:

1.

18. An α-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to hydrochloric acid is 1:1, and its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 12.578, 13.583, 18.151, 19.174, 20.027 and 26.

978.

19. The α-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 18, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 8.647, 11.210, 12.578, 13.583, 18.151, 19.174, 20.027, 26.676 and 26.

978.

20. The α-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 18, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.253, 8.647, 11.210, 12.578, 13.583, 18.151, 19.174, 20.027, 24.105, 25.004, 25.375, 26.676 and 26.

978.

21. A b-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 1, wherein the molar ratio of the compound of formula (I) to hydrochloric acid is 1:1, and its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186 and 20.

074.

22. The b-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 21, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186, 20.074, 20.773, 21.186 and 26.

977.

23. The b-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 21, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 12.613, 15.871, 16.013, 17.839, 18.144, 19.186, 20.074, 20.773, 21.186, 22.512, 24.181, 26.599 and 26.

977.

24. A c-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 1, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 15.515, 17.137, 19.743, 20.471, 21.525, 23.442 and 25.

987.

25. The c-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 24, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 7.353, 13.066, 14.042, 15.515, 17.137, 19.743, 20.471, 21.525, 23.442 and 25.

987.

26. The c-crystal form of the hydrochloride salt of the compound of formula (I) according to claim 24, wherein the X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 6.970, 7.353, 9.527, 13.066, 14.042, 15.515, 17.137, 19.743, 20.471, 21.525, 23.442, 25.987 and 29.

252.

27. A method for preparing the α-crystal form of the hydrochloride salt of the compound of formula (I) according to any one of claims 18-20, the method comprising the following steps: 1) Dissolve the compound of formula (I) in a solvent selected from a mixture of ethanol and diethyl ether, a mixture of butanone and n-hexane, or isopropanol or butanone. 2) Add hydrochloric acid, and crystals will precipitate.

28. A method for preparing the b-crystal form of the hydrochloride salt of the compound of formula (I) according to any one of claims 21-23, the method comprising the following steps: 1) Dissolve the compound shown in formula (I) in a mixed solvent selected from isopropanol, isopropyl ether, and n-hexane, or a mixed solvent of n-hexane and ethyl acetate, or acetonitrile. 2) Add hydrochloric acid, and crystals will precipitate.

29. The crystal form of the maleate salt of the compound of formula (I) according to any one of claims 4-12, and the crystal form of the hydrochloride salt of the compound of formula (I) according to any one of claims 18-26, wherein, The error range of the 2θ angle is ±0.

2.

30. A composition prepared from any one or a mixture thereof of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 1-3, 16-17, or a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 4-12, 15-26, 29, or a pharmaceutically acceptable salt of the compound of formula (I) prepared by the method according to any one of claims 13-14, 27-28.

31. A pharmaceutical composition comprising the following components: i) a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 1-3, 16-17, or a crystal form of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 4-12, 15-26, 29, or any one or a mixture thereof of a pharmaceutically acceptable salt of the compound of formula (I) prepared by the method according to any one of claims 13-14, 27-28; and ii) One or more pharmaceutically acceptable carriers or excipients.

32. A method for preparing a pharmaceutical composition, comprising the step of mixing any one or a mixture thereof with a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 1-3, 16-17, or a crystal form of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 4-12, 15-26, 29, or a crystal form of a pharmaceutically acceptable salt of the compound of formula (I) prepared by the method according to any one of claims 13-14, 27-28, with a pharmaceutically acceptable carrier and an excipient.

33. The use of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 1-3, 16-17, or a crystal form of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 4-12, 15-26, 29, or any crystal form of a pharmaceutically acceptable salt of the compound of formula (I) prepared by the method according to any one of claims 13-14, 27-28, or a mixture thereof, or the use of the composition according to any one of claims 30-31 in the preparation of a medicament for the prevention and / or treatment of estrogen receptor-mediated diseases or conditions.

34. According to the use described in claim 33, the estrogen receptor-mediated disease or condition is cancer.

35. The use according to claim 34, wherein the cancer is breast cancer, ovarian cancer, endometrial cancer, prostate cancer, or uterine cancer.

36. The use according to claim 35, wherein the cancer is breast cancer.

37. The use of any pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 1-3, 16-17, or a crystal form of a pharmaceutically acceptable salt of the compound of formula (I) according to any one of claims 4-12, 15-26, 29, or any crystal form of a pharmaceutically acceptable salt of the compound of formula (I) prepared by the method according to any one of claims 13-14, 27-28, or a mixture thereof, or the use of the composition according to any one of claims 30-31 in the preparation of a medicament for the prevention and / or treatment of cancer, wherein the cancer is breast cancer.

Images