Crystals of naphthofuran-substituted glutarimide compounds, production method and use thereof

Abstract

The present invention discloses crystals of naphthofuran-substituted glutarimide compounds, a method for producing the same, and uses thereof, and more specifically, the present invention discloses crystals of compound (1) and the production thereof. JPEG2025531599000016.jpg3499

Description

[Technical Field]

[0001] This invention claims priority to: CN2022111197111, filing date: September 14, 2022; CN2023111054610, filing date: August 29, 2023.

[0002] The present invention relates to a naphthofuran-substituted glutarimide-based compound crystal, a preparation method thereof, and its use. Specifically, the present invention relates to a crystal of Compound 1 and its preparation. [Background technology]

[0003] Because androgen receptor signaling plays an important role in the pathogenic mechanism of prostate cancer and is known to be involved in the development of other androgen receptor-positive cancers, inhibition of androgen receptor signaling by antiandrogens that antagonize the androgen receptor has been used or proposed for the treatment of prostate cancer.

[0004] The AR gene encodes the androgen receptor protein, whose ligands are primarily testosterone and dihydrotestosterone. This receptor is widely distributed in most organs and tissues of the human body and mediates the biological effects of androgens. The androgen receptor can bind to heat shock proteins (HSPs) in the cytoplasm. When androgens bind to the androgen receptor, the receptor is activated, dissociating the HSPs. The androgen receptor then dimerizes and enters the nucleus, where it binds to androgen response elements (AREs) on DNA, initiating the transcription and expression of a series of genes downstream of the elements, including prostate-specific antigen (PSA), prostatic acid phosphatase (PAP), and the cyclin-dependent kinase (CDK) inhibitor p21WAF1 / CIPI. This ultimately leads to cell differentiation and promotes tissue and organ development. Androgens function to maintain prostate growth and development. In an androgen-free environment, prostate cells spontaneously undergo apoptosis, whereas in an environment with normal androgen levels, prostate cells can continue to grow and differentiate. Therefore, some studies suggest that both excessive secretion of androgens and excessive response of androgen receptors cause uninhibited proliferation of prostate cells and are risk factors for the development and progression of prostate cancer.

[0005] Prostate cancer (PCa) is one of the most commonly diagnosed non-skin cancers among men in the United States and the second leading cause of cancer death, with more than 200,000 new cases and more than 30,000 deaths annually in the United States. Androgen deprivation therapy (ADT) is the standard treatment for advanced PCa. Patients with advanced PCa receive ADT with luteinizing hormone-releasing hormone (LHRH) agonists, LHRH antagonists, or bilateral orchiectomy. Despite an initial response to ADT, disease progression is inevitable, and the cancer manifests as castration-resistant prostate cancer (CRPC). Up to 30% of prostate cancer patients treated with primary radiation or surgery develop metastatic disease within 10 years of primary treatment. Approximately 50,000 patients develop metastatic disease each year, referred to as metastatic CRPC (mCRPC).

[0006] Proteolysis Targeting Chimera (PROTAC) is a technology that utilizes the ubiquitin-proteasome system to target specific proteins and induce their intracellular degradation. The ubiquitin-proteasome system is the primary pathway for intracellular protein degradation and is primarily responsible for removing denatured, mutated, or harmful proteins from cells as a normal physiological function. More than 80% of intracellular protein degradation relies on the ubiquitin-proteasome system. PROTAC utilizes the cell's own protein destruction mechanism to remove specific target proteins in cells.

[0007] The present invention describes compounds, including compositions of matter, that function to recruit endogenous proteins to E3 ubiquitin ligases, such as cereblon (CRBN) E3 ubiquitin ligase, for ubiquitination and subsequent degradation, as well as methods of using the same. In particular, the present invention provides bifunctional or proteolysis-directed chimeric molecules (PROTAC) compounds that have been found to act as modulators of targeted ubiquitination and androgen receptor (AR) degradation. Summary of the Invention

[0008] The present invention provides a type A crystal of Compound 1 shown below, which is characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°.

[0009] [ka]

[0010] In some embodiments of the present invention, the above-mentioned Form A crystal of Compound 1 contains at least 5, 6, 7, or 8 diffraction peaks in a powder X-ray diffraction spectrum at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°.

[0011] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned Type A crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°.

[0012] In some embodiments of the present invention, the above-mentioned Form A crystal of Compound 1 contains at least 10, 11, 12, or 13 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 16.973±0.200°, 17.521±0.200°, 17.858±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

[0013] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned Type A crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 17.521±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

[0014] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned Type A crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 17.521±0.200°, 17.858±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

[0015] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned Type A crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 16.973±0.200°, 17.521±0.200°, 17.858±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

[0016] In some embodiments of the present invention, the above-mentioned Form A crystal of Compound 1 has a powder X-ray diffraction spectrum having characteristic diffraction peaks at the following 2θ angles: 4.358°, 9.530°, 12.135°, 12.784°, 14.166°, 15.597°, 16.622°, 16.973°, 17.521°, 17.858°, 18.617°, 19.495°, 20.531°, 20.956°, 22.271°, 22.815°, 24.711°, 25.955°, 26.976°, 27.682°, 28.728°, 30.119°, 31.329°, 33.598°, and 35.500°.

[0017] In some embodiments of the present invention, the peak positions and relative intensities of the diffraction peaks in the powder X-ray diffraction (XRPD) spectrum of the above-mentioned crystalline form A of Compound 1 are as shown in Table 1.

[0018] [Table 1]

[0019] In some embodiments of the present invention, the above-mentioned crystalline form A of Compound 1 has an XRPD pattern as shown in FIG.

[0020] In some embodiments of the present invention, the above-mentioned Type A crystal of Compound 1 has an endothermic peak onset at 279.6°C ± 3.0°C in its differential scanning calorimetry curve.

[0021] In some embodiments of the present invention, the above-mentioned Type A crystal of Compound 1 has a DSC spectrum as shown in FIG.

[0022] In some embodiments of the present invention, the thermogravimetric analysis curve of the above-mentioned Type A crystal of Compound 1 shows a weight loss of 0.59% at 150.0°C±3.0°C.

[0023] In some embodiments of the present invention, the above-mentioned Type A crystal of Compound 1 has a TGA spectrum as shown in FIG.

[0024] The present invention provides a B-type crystal of Compound 1 shown below, which is characterized in that its powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 16.923±0.200°, 18.394±0.200°, and 21.271±0.200°.

[0025] [ka]

[0026] In some embodiments of the present invention, the above-mentioned type B crystal of Compound 1 comprises at least 5, 6, 7, or 8 diffraction peaks in a powder X-ray diffraction spectrum at the following 2θ angles: 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, and 21.599±0.200°.

[0027] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned B-type crystals of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, and 21.599±0.200°.

[0028] In some embodiments of the present invention, the above-mentioned Type B crystal of Compound 1 comprises at least 10, 11, 12, or 13 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 5.222±0.200°, 10.412±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, 23.951±0.200°, and 25.463±0.200°.

[0029] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned type B crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 5.222±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, and 23.951±0.200°.

[0030] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned type B crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 5.222±0.200°, 10.412±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, and 23.951±0.200°.

[0031] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned type B crystal of Compound 1 has characteristic diffraction peaks at the following 2θ angles: 5.222±0.200°, 10.412±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, 23.951±0.200°, and 25.463±0.200°.

[0032] In some embodiments of the present invention, the powder X-ray diffraction spectrum of the above-mentioned type B crystals has characteristic diffraction peaks at the following 2θ angles: 5.222°, 8.368°, 10.412°, 13.031°, 14.668°, 16.039°, 16.923°, 18.394°, 19.070°, 19.445°, 20.915°, 21.271°, 21.599°, 23.591°, 25.463°, 27.635°, and 31.559°.

[0033] In some embodiments of the present invention, the peak positions and relative intensities of the diffraction peaks in the powder X-ray diffraction (XRPD) spectrum of the above-mentioned B-type crystals of Compound 1 are as shown in Table 2.

[0034] [Table 2]

[0035] In some embodiments of the present invention, the Type B crystal of the above solvate has an XRPD spectrum as shown in FIG.

[0036] In some embodiments of the present invention, the Type B crystals of the solvate have an endothermic peak onset at 290.2°C ± 3.0°C in their differential scanning calorimetry curve (DSC).

[0037] In some embodiments of the present invention, the DSC spectrum of the Type B crystal of the above solvate is as shown in FIG.

[0038] In some embodiments of the present invention, the Type B crystal of the solvate exhibits a weight loss of 1.75% at 150°C ± 3.0°C in a thermogravimetric analysis curve (TGA).

[0039] In some embodiments of the present invention, the type B crystal of the above-mentioned solvate has a TGA spectrum as shown in FIG.

[0040] In some embodiments of the present invention, the above-mentioned type B crystals of Compound 1 may exist in the form of solvate crystals.

[0041] The present invention also provides a method for producing the above-mentioned type B crystals of Compound 1, which comprises precipitating type A crystals of Compound 1 in methyl tert-butyl ether, and further comprising a separation step.

[0042] In some embodiments of the present invention, the B-type crystal of Compound 1 of the present invention is (1) dissolving compound 1 in N,N-dimethylacetamide under ultrasonic conditions; (2) filtering and then adding methyl tert-butyl ether; (3) stirring at low temperature for 48 hours and then returning to room temperature; (4) Filtration process Manufactured by

[0043] In some embodiments of the present invention, the low temperature referred to in step (3) above is preferably 5°C.

[0044] The present invention also provides a method for treating a coronavirus infection, comprising administering a therapeutically effective amount of Form A crystals or Form B crystals of Compound 1 of the present invention to an individual in need thereof.

[0045] The present invention also provides use of the above-mentioned type A crystals or type B crystals of Compound 1 in the manufacture of a medicament for treating a disease associated with androgen protein degradation by targeting a chimera.

[0046] In some embodiments of the present invention, the disease associated with androgen protein degradation targeted by the chimeras described above is prostate cancer.

[0047] [Technical Effects] The A-type and B-type crystals of Compound 1 of the present invention have stable properties, low hygroscopicity, and good potential for drug discovery.

[0048] [Definitions and Explanations] Unless otherwise stated, the following terms and phrases used herein have the following meanings. A particular phrase or term, unless specifically defined, should be understood to have its ordinary definition, not to be indefinite or unclear. When a trade name appears in this specification, it refers to the corresponding product or its active ingredient.

[0049] It is well known in the field of crystallography that for any given crystalline form, the relative intensities of diffraction peaks can vary due to factors such as preferred orientation due to crystal morphology. While the influence of preferred orientation can alter peak intensities, the positions of the diffraction peaks of a crystal cannot. Furthermore, it is well known in the field of crystallography that slight errors in peak positions can exist for any given crystal. For example, peak positions can shift due to temperature changes during sample analysis, sample movement, or instrument calibration, resulting in a measurement error of approximately ±0.2 degrees for the 2θ values. Therefore, it is well known to those skilled in the art that this error must be taken into account when determining the structure of each crystal.

[0050] DSC measures the transition temperatures that occur when heat is absorbed or released due to changes in crystalline structure or crystalline melting. For the same crystal of the same compound, the error between the thermal transition temperature and melting point in consecutive analyses is usually within approximately 5°C or 3°C. If a compound has a specific DSC peak or melting point, this means that the DSC peak or melting point is within ±5°C or ±3°C. DSC provides an auxiliary method for distinguishing different crystals. Different crystals can be identified based on their different transition temperature characteristics. It should be noted that for mixtures, the DSC peak or melting point may vary over a wider range. Furthermore, because the melting process of a substance involves decomposition, the melting temperature is related to the heating rate.

[0051] For the same crystal, the weight loss temperature in TGA may vary depending on factors such as the measuring instrument, measurement method / conditions, etc. For any particular crystal, there may be an error in the weight loss temperature, which may be about ±5°C or about ±3°C.

[0052] It should be noted that in the preparation of drug crystals, when drug molecules and solvent molecules come into contact with each other, external and internal conditions can cause the solvent molecules and compound molecules to form a eutectic, which is difficult to avoid remaining in the solid material, resulting in the formation of solvates, specifically including stoichiometric solvates and non-stoichiometric solvates. Both of these solvates are included within the scope of the present invention.

[0053] The term "pharmaceutically acceptable excipient" refers to an inert substance that is co-administered with an active ingredient and facilitates the administration of the active ingredient, and includes, but is not limited to, any flow aid, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, disintegrant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that is approved by the State Food and Drug Administration of China for use in humans or animals (e.g., livestock).

[0054] The term "crystalline composition" refers to a mixture of a crystal of Compound 1 of the present invention with other crystalline or amorphous substances of the compound, or other impurities. For example, a crystalline composition of Form A crystal of Compound 1 contains, in addition to Form A crystal of Compound 1, other crystalline or amorphous substances of Compound 1, or other impurities.

[0055] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or salts thereof with a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of the compounds of the present invention to an organism.

[0056] The therapeutic dosage of a compound of the invention may be determined based on, for example, the particular therapeutic application, the manner in which the compound is administered, the patient's health condition, and the judgment of the prescribing physician. The proportion or concentration of a compound of the invention in a pharmaceutical composition is not fixed and will depend upon various factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration.

[0057] The term "treatment" means administering a compound or formulation according to the present invention to ameliorate or eliminate a disease or one or more symptoms associated with said disease; (i) inhibiting a disease or disease state, i.e., arresting its development; (ii) alleviating a disease or disease state, i.e., eliminating the disease or disease state This includes:

[0058] The term "therapeutically effective amount" refers to an amount of a compound of the present invention that (i) treats a particular disease, condition, or disorder, (ii) reduces, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder described herein. The amount of a compound of the present invention that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the method of administration, and the age of the mammal being treated, but can be determined routinely by one of ordinary skill in the art in light of their own knowledge and the present invention.

[0059] Unless otherwise required by the present invention, throughout this specification and the claims that follow, the word "comprise" and its English variants such as "comprises" and "comprising" are to be interpreted in an open-ended and inclusive sense, i.e., "including but not limited to."

[0060] Throughout this specification, a reference to "one embodiment" or "an embodiment" or "another embodiment" or "in some embodiments" means that at least one embodiment includes the particular referenced element, structure, or feature described in that embodiment. Thus, the appearances of the phrases "in one embodiment" or "in an embodiment" or "in another embodiment" or "in some embodiments" in various places throughout this specification do not necessarily all refer to the same embodiment. Furthermore, particular elements, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0061] As used in this specification and the appended claims, the singular form "a" (corresponding to the English words "a," "an," and "the") should be understood to include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a reaction including a "catalyst" includes one catalyst, or two or more catalysts. It should further be understood that the term "or" is generally used in its sense to include "and / or" unless the context clearly dictates otherwise.

[0062] The intermediate compounds of the present invention can be prepared by various synthetic methods familiar to those skilled in the art, including the specific embodiments listed below, embodiments in combination with other chemical synthetic methods, and equivalent alternative methods familiar to those skilled in the art, and preferred embodiments include, but are not limited to, the examples of the present invention.

[0063] The chemical reactions of specific embodiments of the present invention are completed in suitable solvents, which should be suitable for the chemical reactions of the present invention and the reagents and materials required therefor. To obtain the compounds of the present invention, those skilled in the art may need to modify or select synthetic steps or reaction schemes based on existing embodiments.

[0064] [ka]

[0065] The present invention will be specifically described below with reference to examples, but these examples do not limit the present invention in any way.

[0066] All solvents used in this invention are commercially available and may be used as is without further purification.

[0067] Compounds were named according to their usual names in the art or ChemDraw® software, and for commercially available compounds the manufacturer's catalogue names were used.

[0068] The present invention uses the following abbreviations: Boc stands for tert-butoxycarbonyl; THF stands for tetrahydrofuran; DMSO stands for dimethyl sulfoxide; EA or EtOAc stands for ethyl acetate; MeOH stands for methanol; IPA stands for isopropanol; ACN stands for acetonitrile; DMAC stands for N,N-dimethylacetamide; RT stands for room temperature.

[0069] Instrumentation and analytical methods

[0070] 1.1 X-ray powder diffractometer (XRPD) method of the present invention Instrument model: PANalytacal powder X-ray diffraction analyzer Measurement method: Approximately 10 to 20 mg of sample is used for XRPD detection. The detailed XRPD parameters are as follows: X-ray tube: Cu, kα, (λ=540598Å, λ=1.544426Å). Tube voltage: 40 kV, tube current: 40 mA Divergence slit: 1 / 8° Scanning mode: Continuous Scanning range (°2 Theta): 3~40 Scan time per step (s): 46.7 Step width: 0.0263 seconds Test time: ~5 min

[0071] 1.2 Differential Scanning Calorimeter (DSC) Method of the Present Invention Instrument model: TA 2500 differential scanning calorimeter Measurement method: A sample (approximately 1 mg) is placed in a DSC aluminum crucible and heated from 30°C (room temperature) to 300°C (or 350°C) at a heating rate of 10°C / min under conditions of 50 mL / min N2.

[0072] 1.3 Thermogravimetric Analysis (TGA) Method of the Present Invention Instrument model: TA 5500 Thermogravimetric Analyzer Measurement method: A sample (2-5 mg) is placed in a TGA platinum crucible and heated from room temperature to 350°C at a heating rate of 10°C / min under conditions of 25 mL / min N2, or until the weight is reduced by 20%. or: Thermogravimetric analyzer TGA550 Measurement method: A sample (5-10 mg) is placed on the built-in aluminum plate of a TGA platinum crucible and heated from room temperature to 300°C at a heating rate of 10°C / min under conditions of 60 mL / min N2.

[0073] 1.4 Dynamic Vapor Sorption (DVS) Method of the Present Application Instrument Model: SEM Advantage-1 Dynamic Vapor Sorption Apparatus Measurement conditions: The sample (10-20 mg) is measured in a DVS sample pan. The detailed DVS parameters are as follows: Temperature: 25℃ Balance: dm / dt = 0.01% / min (min: 10 min, max: 180 min) Drying: 120 min at 0% RH RH(%) test gradient: 10% RH (%) test gradient range: 0%~95%~0% [Brief explanation of the drawings]

[0074] [Figure 1] 1 is an XRPD spectrum of Cu-Kα radiation of the A-type crystal of Compound 1. [Figure 2] 1 is a DSC spectrum of type A crystal of Compound 1. [Figure 3] 1 is a TGA spectrum of type A crystals of Compound 1. [Figure 4] 1 is an XRPD spectrum of Cu-Kα radiation of B-type crystals of Compound 1. [Figure 5] 1 is a DSC spectrum of type B crystals of Compound 1. [Figure 6] 1 is a TGA spectrum of type B crystals of Compound 1. [Figure 7] 1 is a DVS spectrum of type A crystal of Compound 1. [Figure 8] 1 is a DVS spectrum of type B crystal of Compound 1. DETAILED DESCRIPTION OF THE INVENTION

[0075] The present application will be specifically described below by way of examples, but is not intended to limit the present invention in any way. The present application has been described in detail herein, and specific embodiments thereof have also been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made in the specific embodiments of the present application without departing from the spirit and scope of the present application.

[0076] Example 1: Preparation of Type A Crystals of Compound 1

[0077] [ka]

[0078] Step 1: Synthesis of Compound 1-2 Compound 1-1 (27.86 g, 83.63 mmol), N-Boc-piperazine (23.36 g, 125.44 mmol), tris(dibenzylideneacetone)dipalladium (7.66 g, 8.36 mmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (7.10 g, 16.73 mmol), and potassium phosphate (53.25 g, 250.88 mmol) were added to HO (50 mL) and 1,4-dioxane (500 mL) at room temperature under nitrogen gas protection. The reaction mixture was slowly heated to 100 °C and stirred for 12 h. After completion of the reaction, the reaction mixture was poured into water (500 mL) and extracted with ethyl acetate (500 mL × 2). The combined organic phase was washed with saturated brine (500 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 93 / 7, volume ratio) to obtain compound 1-2. (Eluent: petroleum ether / ethyl acetate = 1 / 0 to 93 / 7, volume ratio) to obtain compound 1-2. 1H NMR (400MHz, CDCl3) δ: 8.21 (d, J=9.3 Hz, 1H), 7.97 (d, J=8.3 Hz, 1H), 7.75 (s, 1H), 7.65 (d, J=9.3 Hz, 1H), 7.51 (t, J=7.9 Hz, 1H), 7.12 (d, J=7.3 Hz, 1H), 4.26-4.06 (m, 6H) 3.44 - 2.70 (m, 6H), 1.52 (s, 9H), 1.27 (t, J=7.1 Hz, 3H)

[0079] Step 2: Synthesis of Compounds 1-3 Compound 1-2 (25 g, 57.01 mmol), acrylamide (6.08 g, 85.52 mmol, 5.90 mL), and potassium tert-butoxide (9.60 g, 85.52 mmol) were dissolved in N,N-dimethylformamide (300 mL) at room temperature under nitrogen gas protection, and the reaction mixture was stirred at 0-5°C for 2 h. After completion of the reaction, the reaction mixture was poured into saturated ammonium chloride solution (400 mL), filtered, and the cake was washed with water (250 mL) and spin-dried. The resulting crude product was slurried in methanol (100 mL, 25°C, 12 h) and purified to give compound 1-3. 1 H NMR (400 MHz, DMSO_d6) δ: 10.95 (s, 1H), 8.20 (d, J = 9.2 Hz, 1H), 8.00 (s, 1H), 7.90 (m, 1H), 7.78 (d, J = 9.2 Hz, 1H), 7.52 (m, 1H), 7.20 (d, J = 7.6 Hz, 1H), 4.66 (m, 1H), 3.29 - 3.12 (m, 4H), 3.03 - 2.57 (m, 5H), 2.45 - 2.21 (m, 3H), 1.45 (s, 9H).

[0080] Step 3: Synthesis of the hydrochloride salts of compounds 1-4 Compound 1-3 (21.5 g, 46.38 mmol) was dissolved in ethyl acetate (500 mL) at room temperature, and a solution of hydrochloric acid in ethyl acetate (4 M, 250 mL) was added. The reaction mixture was stirred at room temperature for 48 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by slurrying with ethyl acetate (30 mL) at room temperature to obtain the hydrochloride salt of compound 1-4. 1 H NMR (400 MHz, DMSO_d6) δ: 10.95 (s, 1H), 9.54 (br, 1H), 8.19 (d, J = 9.2 Hz, 1H), 8.05 - 7.89 (m, 2H), 7.80 (d, J = 9.2 Hz, 1H), 7.55 (m, 1H), 7.22 (d, J = 7.6 Hz, 1H), 4.74 - 4.62 (m, 1H), 3.44 - 3.19 (m, 8H), 2.85 - 2.95 (m, 1H), 2.60 - 2.66 (m, 1H), 2.47 - 2.36 (m, 1H), 2.20 - 2.30 (m, 1H).

[0081] Step 4: Synthesis of Compounds 1-6 Compound 1-4 hydrochloride (16.35 g, 40.89 mmol), compound 1-5 (16.0 g, 34.12 mmol), and sodium triacetoxyborohydride (10.83 g, 51.11 mmol) were dissolved in dichloromethane (1500 mL) at room temperature under nitrogen gas protection. The reaction mixture was stirred at room temperature for 48 hours, and sodium acetate (5.59 g, 68.15 mmol) was added. The reaction mixture was continued to stir for 24 hours. After completion of the reaction, the mixture was quenched with water (500 mL), separated, and the organic phase was washed with saturated brine (500 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was separated by column chromatography (eluent: ethyl acetate / methanol = 95 / 5, volume ratio). The crude product was slurried with ethyl acetate (50 mL) at room temperature to give compound 1-6. MS-ESI m / z: 815.2 [M+H] + . 1H NMR (400 MHz, DMSO_d6) δ: 10.95 (s, 1H), 8.59 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 9.2 Hz, 1H), 7.99 (s, 1H), 7.92 - 7.73 (m, 4H), 7.53 - 7.40 (m, 1H), 7.43 - 7.29 (m, 2H), 7.24 - 7.09 (m, 2H), 4.72 - 4.60 (m, 1H), 4.59 - 4.45 (m, 3H), 3.93 - 3.80 (m, 1H), 3.24 - 2.96 (m, 6H), 2.95 - 2.82 (m, 2H), 2.76 - 2.58 (m, 3H), 2.48 - 2.18 (m, 5H), 2.16 - 2.06 (m, 2H), 1.99 - 1.79 (m, 5H), 1.72 - 1.44 (m, 4H), 1.25 - 1.07 (m, 2H).

[0082] Step 5: Chiral HPLC separation Compounds 1-6 were separated by chiral HPLC (column type: DAICEL CHIRALPAK IE (250 mm × 30 mm, 10 μm); mobile phase: A (IPA), B (ACN), B%: 50% to 100%; flow rate: 80 mL / min) to obtain type A crystals of compound 1 (the XRPD spectrum of which is shown in Figure 1).

[0083] Analytical method: Chromatography column: Chiralpak IA 100 × 4.6 mm, 3 μm; Mobile phase: A: n-hexane (0.1% diethylamine); B: isopropanol:acetonitrile = 2:1, A:B = 40:60; Column temperature: 35 °C; Wavelength: 220 nm.

[0084] Compound 1 (first main peak, retention time: 4.857 min): 1H NMR (400 MHz, DMSO_d6) δ: 10.95 (s, 1H), 8.60 (d, J = 8.4 Hz, 1H), 8.16 (d, J = 9.2 Hz, 1H), 8.00 (s, 1H), 7.91 - 7.74 (m, 4H), 7.53 - 7.40 (m, 1H), 7.43 - 7.30 (m, 2H), 7.23 - 7.09 (m, 2H), 4.72 - 4.60 (m, 1H), 4.60 - 4.45 (m, 3H), 3.94 - 3.80 (m, 1H), 3.24 - 2.96 (m, 6H), 2.95 - 2.79 (m, 2H), 2.76 - 2.58 (m, 3H), 2.45 - 2.18 (m, 5H), 2.16 - 2.06 (m, 2H), 2.03 - 1.82 (m, 5H), 1.73 - 1.44 (m, 4H), 1.24 - 1.09 (m, 2H).

[0085] Example 2: Preparation of B-type crystals of Compound 1 At room temperature, type A crystals of compound 1 (3.35 g) were dissolved in DMAC (100 mL), sonicated to promote dissolution, and filtered. After filtration, methyl tert-butyl ether (1000 mL) was added, cooled to 5°C, and stirred for 48 hours to precipitate a solid. The mixture was returned to room temperature and filtered. The resulting cake was type B crystals of compound 1 (the XRPD spectrum of which is shown in Figure 4).

[0086] Analytical method: Chromatography column: Chiralpak IA 100 × 4.6 mm, 3 μm; Mobile phase: A: n-hexane (0.1% diethylamine); B: isopropanol:acetonitrile = 2:1, A:B = 40:60; Column temperature: 35 °C; Wavelength: 220 nm; Retention time: 4.856 min; MS-ESI m / z: 815.3 [M + H] + .

[0087] Experimental Example 1: Hygroscopicity study of the A-type crystals of Compound 1 Test materials: SEM Advantage-1 Dynamic Vapor Sorption Apparatus.

[0088] Experimental Method: 10 to 20 mg of type A crystals of Compound 1 were placed in a DVS sample pan and tested.

[0089] Test Results: The DVS spectrum of the A-type crystal of Compound 1 is shown in FIG.

[0090] Testing Conclusion: The hygroscopic weight gain of the A-type crystal of Compound 1 at 25°C and 80% RH was 0.3789%, indicating that the crystal was slightly hygroscopic, and the crystal did not change before and after DVS.

[0091] Experimental Example 2: Hygroscopicity study of B-type crystals of Compound 1 Test materials: SEM Advantage-1 Dynamic Vapor Sorption Apparatus. Experimental Method: 10 to 20 mg of type B crystals of Compound 1 were placed in a DVS sample pan and tested.

[0092] Test Results: The DVS spectrum of the B-type crystal of Compound 1 was shown in FIG. 8, with ΔW=0.555%.

[0093] Testing Conclusion: The hygroscopic weight gain of the B-type crystal of Compound 1 at 25°C and 80% RH was 0.555%, indicating that the crystal was slightly hygroscopic, and the crystal did not change before and after DVS.

[0094] Experimental Example 3: Interconversion relationship between A-type crystals and B-type crystals of Compound 1 in different solvents The inter-transformation relationship between two amorphous crystals (crystal type A and crystal type B) was investigated through a competitive suspension test.

[0095] Approximately 20 mg of a sample of type A crystals was weighed and added to 1 mL of solvent, stirred at the corresponding temperature, and filtered. The filtrate was transferred to an HPLC vial containing type A and type B crystals, suspended, stirred, and centrifuged at the corresponding temperature, and the wet sample XRPD was measured. The results are summarized in Table 3.

[0096] [Table 3]

[0097] Experimental conclusion: The results show that type B crystals can be obtained in each solvent system at different temperatures.

[0098] Experimental Example 4: Solid-state stability experiment of B-type crystals of Compound 1 under accelerated conditions The stability of type B crystals of compound 1 was determined under accelerated conditions of high temperature and high humidity (40°C / relative humidity 75%, sealed) according to the "Guiding Principles for Stability Testing of Drug Substances and Preparations" (Chinese Pharmacopoeia 2020 Edition, Part 4, 0512).

[0099] Approximately 1.2 g of B-type crystals of Compound 1 were weighed and placed in two low-density polyethylene bags. Each low-density polyethylene bag was sealed with a zipper and then placed in an aluminum foil bag and heat-sealed. Samples were taken in January, February, and March and the results were compared with the initial results on day 0. The experimental results are shown in Table 4.

[0100] [Table 4]

[0101] Conclusion: Form B crystals of compound 1 have good stability under accelerated conditions of 40°C / 75% relative humidity.

[0102] Experimental Example 5: Solid-state stability experiment of B-type crystals of Compound 1 under long-term conditions The stability of type B crystals of compound 1 under long-term conditions (25°C / relative humidity 60%, sealed) was determined according to the "Guiding Principles for Stability Testing of Drug Substances and Preparations" (Chinese Pharmacopoeia 2020 Edition, Part 4, 0512).

[0103] Approximately 1.2 g of B-type crystals of Compound 1 were weighed and placed in two low-density polyethylene bags. Each low-density polyethylene bag was sealed with a zipper and then placed in an aluminum foil bag and heat-sealed. Samples were taken and tested at month 3, and the test results were compared with the initial test results on day 0. The experimental results are shown in Table 5.

[0104] [Table 5]

[0105] Conclusion: The B-type crystals of compound 1 have good stability under long-term conditions of 25°C / 60% relative humidity.

[0106] Biological Testing Test Example 1: Study of the anti-cell proliferation effect of Compound 1 in AR F877L mutant LNCap cells using in vitro cell activity experiments Experimental Objective: In this experiment, the inhibitory effect of compound 1 on cell proliferation was investigated by detecting the effect of compound 1 on in vitro cell activity in AR F877L mutant LNCap cells.

[0107] Test materials: 1. Cell lines and culture methods

[0108] [Table 6]

[0109] 2. Media and Reagents

[0110] [Table 7]

[0111] 3. Experimental Equipment 1) Greiner CELLSTAR 96-well plate, flat bottom, black plate (with lid and clear bottom), #655090 2) Promega CellTiter-Glo Luminescent Cell Activity Detection Kit (Promega-G7573). 3) 2104 EnVision plate reader, PerkinElmer.

[0112] Experimental Method: 1. Cell Culture: Tumor cells were cultured in an incubator at 37°C and 5% CO2 according to the culture conditions shown in Table 6. They were passaged periodically, and cells in the logarithmic growth phase were collected for plating. 1) Cells were stained with trypan blue and viable cells were counted. 2) The cell concentration was adjusted to an appropriate density of 5000 cells / well. 3) The cell suspension was added to the culture plate at 90 μL / well, and cell-free medium was added to blank control wells. 4) The culture plate was incubated overnight in an incubator at 37°C, 5% CO2, and 100% relative humidity.

[0113] 3. Preparation of Compound Storage Plates and Activity Detection 1) Preparation of a 400-fold compound stock plate: Compounds were diluted in DMSO from highest to lowest concentration gradient. 2) Preparation of 10X compound working solution: 78 μL of cell culture medium was added to a V-bottom 96-well plate, and 2 μL of compound from the 400X compound stock plate was pipetted into the cell culture medium in the 96-well plate. 2 μL of DMSO was added to the solvent control and blank control. Compound or DMSO was added and mixed evenly with a pipette. 3) Administration: 10 μL of 10X compound working solution was taken and added to the cell culture plate. 10 μL of DMSO-cell medium mixed solution was added to the solvent control and blank control. The final concentration of DMSO was 0.25%. 4) The 96-well cell plate was returned to the incubator and cultured for 144 hours. 5) CellTiter-Glo buffer was dissolved and left at room temperature. 6) The CellTiter-Glo substrate was left at room temperature. 7) CellTiter-Glo buffer was added to the bottle of CellTiter-Glo substrate to dissolve the substrate, producing a CellTiter-Glo working solution. 8) Gently vortex to dissolve completely. 9) The cell culture plate was removed and left for 30 minutes to equilibrate to room temperature. 10) 50 μL of CellTiter-Glo working solution was added to each well (equivalent to half the volume of the cell culture medium in each well). The cell plate was wrapped in aluminum foil to protect it from light. 11) The culture plate was shaken on an orbital shaker for 2 minutes to induce cell lysis. 12) The culture plate was left at room temperature for 10 minutes to stabilize the luminescence signal. 13) Luminescent signals were detected using a 2104 EnVision plate reader.

[0114] 4. Data Analysis The inhibition rate (IR) of the test compound was calculated using the following formula: IR (%) = (1-(RLU solvent control - RLU compound) / (RLU solvent control - RLU blank control) x 100%. The inhibition rates of different concentrations of the compound were calculated using Excel, and the inhibition curves were plotted using GraphPad Prism software, showing the minimum inhibition rate, maximum inhibition rate, and IC 50 The relevant parameters were calculated, including

[0115] 5. Experimental Results

[0116] [Table 8]

[0117] Conclusion: Compound 1 of the present invention exhibits excellent cell proliferation inhibitory effect in AR F877L mutant LNCaP cells.

Claims

1. A type A crystal of Compound 1, characterized in that the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°. 【Chemical 1】

2. The A-type crystal of compound 1 according to claim 1, characterized in that the A-type crystal contains at least 5, 6, 7, or 8 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°.

3. The A-type crystal of Compound 1 according to claim 1, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 17.521±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, and 22.271±0.200°.

4. The type A crystal of compound 1 according to claim 1, characterized in that the type A crystal contains at least 10, 11, 12, or 13 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 16.973±0.200°, 17.521±0.200°, 17.858±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

5. The A-type crystal of Compound 1 according to claim 1, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 9.530±0.200°, 12.135±0.200°, 12.784±0.200°, 16.622±0.200°, 16.973±0.200°, 17.521±0.200°, 17.858±0.200°, 18.617±0.200°, 19.495±0.200°, 20.531±0.200°, 20.956±0.200°, 22.271±0.200°, and 22.815±0.200°.

6. 2. The A-type crystal of Compound 1 according to claim 1, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 4.358°, 9.530°, 12.135°, 12.784°, 14.166°, 15.597°, 16.622°, 16.973°, 17.521°, 17.858°, 18.617°, 19.495°, 20.531°, 20.956°, 22.271°, 22.815°, 24.711°, 25.955°, 26.976°, 27.682°, 28.728°, 30.119°, 31.329°, 33.598°, and 35.500°.

7. 2. The A-type crystal of Compound 1 according to claim 1, characterized in that the powder X-ray diffraction spectrum is as shown in FIG.

8. 2. The A-type crystal of Compound 1 according to claim 1, characterized in that the differential scanning calorimetry curve has an endothermic peak starting point at 279.6°C ± 3.0°C.

9. 2. The A-type crystal of compound 1 according to claim 1, whose differential scanning calorimetry curve is as shown in FIG.

10. The A-type crystal of Compound 1 according to claim 1, characterized in that the thermogravimetric analysis curve shows a weight loss of 0.59% at 150.0°C ± 3°C.

11. The A-type crystal of compound 1 according to claim 1, having a thermogravimetric analysis curve as shown in Figure 6.

12. A B-type crystal of Compound 1, characterized in that the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 10.655±0.200°, 11.988±0.200°, 16.055±0.200°, 18.356±0.200°, and 20.083±0.200°. 【Chemistry 2】

13. The B-type crystal of compound 1 according to claim 12, characterized in that the B-type crystal contains at least 5, 6, 7, or 8 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, and 21.599±0.200°.

14. The B-type crystal of compound 1 according to claim 12, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, and 21.599±0.200°.

15. The B-type crystal of compound 1 according to claim 12, characterized in that the B-type crystal contains at least 10, 11, 12, or 13 diffraction peaks selected from the following 2θ angles in a powder X-ray diffraction spectrum: 5.222±0.200°, 10.412±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, 23.951±0.200°, and 25.463±0.200°.

16. 13. The B-type crystal of Compound 1 according to claim 12, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 5.222±0.200°, 10.412±0.200°, 14.668±0.200°, 16.039±0.200°, 16.923±0.200°, 18.394±0.200°, 19.070±0.200°, 19.445±0.200°, 20.915±0.200°, 21.271±0.200°, 21.599±0.200°, 23.951±0.200°, and 25.463±0.200°.

17. 13. The B-type crystal of Compound 1 according to claim 12, wherein the powder X-ray diffraction spectrum has characteristic diffraction peaks at the following 2θ angles: 5.222°, 8.368°, 10.412°, 13.031°, 14.668°, 16.039°, 16.923°, 18.394°, 19.070°, 19.445°, 20.915°, 21.271°, 21.599°, 23.591°, 25.463°, 27.635°, and 31.559°.

18. 13. The B-type crystal of compound 1 according to claim 12, characterized in that the powder X-ray diffraction spectrum is as shown in Figure 4.

19. The B-type crystal of compound 1 according to claim 12, characterized in that the differential scanning calorimetry curve has an endothermic peak at 290.2°C ± 3.0°C.

20. 13. The B-type crystal of compound 1 according to claim 12, having a differential scanning calorimetry curve as shown in FIG.

21. The B-type crystal of Compound 1 according to claim 12, characterized in that the thermogravimetric analysis curve shows a weight loss of 1.75% at 150.0°C ± 3°C.

22. The B-type crystal of compound 1 according to claim 12, having a thermogravimetric analysis curve as shown in Figure 6.

23. Use of type A crystals of compound 1 according to claims 1 to 11 or type B crystals of compound 1 according to claims 12 to 22 in the manufacture of a medicament for treating a disease associated with androgen protein degradation targeting a chimera.

24. 24. The use according to claim 23, characterized in that the disease associated with androgen protein degradation targeted by the chimera is prostate cancer.

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