Amorphous, crystalline, methods of preparation and uses of dioxopiperidine compounds or pharmaceutically acceptable salts thereof

Abstract

The present disclosure relates to amorphous, crystalline, preparation method and use of dioxopiperidine compounds or pharmaceutically acceptable salts thereof. In particular, the present disclosure relates to amorphous and crystalline of compounds of formula I or pharmaceutically acceptable salts thereof, which have good physicochemical properties.

Description

[0001] This application claims priority to Chinese patent application 2023105921952, filed on May 24, 2023. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This disclosure pertains to the pharmaceutical field and relates to amorphous forms, crystals, preparation methods, and uses of dioxopiperidine compounds or their pharmaceutically acceptable salts. Background Technology

[0003] PROTAC (PROteolysis TArgeting Chimera) is a hybrid bifunctional small molecule compound. Its structure contains two different ligands: one is a ubiquitin ligase E3 ligand, and the other is a target protein-binding ligand, linked by a linker unit. PROTAC brings the target protein and the intracellular ubiquitin ligase E3 closer together, forming a target protein-PROTAC-E3 ternary complex. The E3 ubiquitin ligase then tags the target protein with a ubiquitinated protein tag, subsequently activating the cell's powerful ubiquitination-proteasome system to specifically degrade the target protein, thereby inhibiting the corresponding protein signaling pathway (Cell Biochem Funct. 2019, 37, 21-30). Compared with traditional small molecule inhibitors, PROTACs exhibit unique advantages: 1) PROTACs do not require prolonged and high-intensity binding to the target protein, and the degradation process of the target protein is similar to a catalytic reaction, allowing for cyclic binding and degradation of the target protein, thereby reducing systemic drug exposure and minimizing toxic side effects; 2) The target protein needs to be resynthesized after degradation to restore its function, therefore, degradation of the target protein shows a more efficient and durable anti-tumor effect than inhibition of its activity, and does not lead to drug resistance due to target protein mutations; 3) PROTACs also have therapeutic potential for targets currently considered untreatable, such as transcription factors, scaffold proteins, and regulatory proteins.

[0004] The discovery of CRBN-type E3 ligase ligands is related to research on the mechanism of action of thalidomide. In 2010, scientists discovered cereblon, a thalidomide-binding protein, during their research on thalidomide toxicity (Science 2010, 327, 1345). Cerebellon is part of the E3 ubiquitin ligase protein complex and acts as a substrate receptor selectively on ubiquitinated proteins. This study indicated that the binding of thalidomide to cereblon in vivo may be the cause of thalidomide's teratogenicity. Subsequent studies found that this compound and related structures can be used as anti-inflammatory, anti-angiogenic, and anticancer agents. Further modifications to the thalidomide structure, resulting in lenalidomide and pomalidomide, have significantly improved safety and reduced teratogenic effects. Lenalidomide was approved for marketing by the FDA in 2006. Two groundbreaking papers published in Science in 2014 pointed out that lenalidomide works by degrading two specific B cell transcription factors, Ikaros family zinc finger structural proteins 1 and 3 (IKZF1 and IKZF3). This further revealed that the thalidomide structure may play a further role in degrading target proteins by binding to the E3 ubiquitin ligase protein complex of cerebellar proteins (Science, 2014, 343, 301; Science, 2014, 343, 305).

[0005] The androgen receptor (AR) is a ligand-dependent trans-transcriptional regulatory protein belonging to the nuclear receptor superfamily and primarily located in the cell nucleus. Unligand-bound ARs bind to heat shock proteins (HSPs); upon ligand binding, ARs undergo a conformational change, dissociating from HSPs and exhibiting increased affinity for DNA (AR activation). Activated ARs, in dimer form, bind to specific DNA sequences in the cell nucleus—androgen response elements (AREs)—and interact with other transcription factors, thereby regulating the expression of related genes and producing biological effects. Studies have shown that abnormalities in the AR signaling pathway are closely related to the occurrence and development of diseases such as prostate cancer, benign prostatic hyperplasia, Kennedy's disease, male infertility, androgen insensitivity, and male breast cancer.

[0006] Prostate cancer is one of the most common malignant tumors. Statistics show that in 2018, there were nearly 1.3 million new cases and 359,000 deaths worldwide, accounting for 13.5% of male malignant tumor incidence, ranking second; and 6.7% of male malignant tumor mortality, ranking fifth. Several AR antagonists have been approved for marketing and successfully used in the treatment of castration-resistant prostate cancer, becoming a major treatment method for prostate cancer. However, most patients develop drug resistance after 0.5-2 years of treatment, leading to disease progression. In some drug-resistant patients, cancer cell growth still depends on the AR signaling pathway.

[0007] PCT / CN2022 / 134333 discloses the structure of a class of compounds for regulating the ubiquitination and degradation of androgen receptor (AR) proteins, chemically named 2-chloro-4-((S)-8-(4-(4-((4-(3-((((S)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile, having the structure shown in Formula I. The entire text is incorporated herein by reference.

[0008] Summary of the Invention

[0009] In a first aspect, this disclosure provides amorphous forms of the compounds represented by Formula I.

[0010]

[0011] In some embodiments, the amorphous form of the compound represented by Formula I has an X-ray powder diffraction pattern without sharp peaks, preferably with non-sharp peaks, and more preferably, its X-ray powder diffraction pattern is as follows: Figure 1 As shown.

[0012] In some embodiments, the amorphous form of the compound shown in Formula I has an endothermic peak value of 122.02 ± 3 °C as shown in its DSC spectrum.

[0013] In some embodiments, the amorphous form of the compound shown in Formula I has a thermogravimetric analysis (TGA) spectrum showing a weight loss of about 5.06% at 33–170 °C.

[0014] This disclosure also provides a method for preparing an amorphous form of the compound shown in Formula I, comprising: mixing the compound shown in Formula I with solvent A, crystallizing, and obtaining an amorphous form. In some embodiments, the solvent A is selected from one or more of water, methanol, ethanol, isopropanol, n-propanol, isoamyl alcohol, isopropyl acetate, methyl tert-butyl ether, n-heptane, cyclohexane, and toluene.

[0015] This disclosure also provides a method for preparing an amorphous form of the compound shown in Formula I, comprising: mixing the compound shown in Formula I with solvent B, adding an antisolvent, and crystallizing to obtain the amorphous form. In some embodiments, the solvent B is selected from one or more of dimethyl sulfoxide, acetone, acetonitrile, tetrahydrofuran, and dichloromethane; and the antisolvent is selected from one or more of water, ethanol, n-heptane, or isopropyl ether.

[0016] In a second aspect, this disclosure also provides a pharmaceutically acceptable salt of the compound represented by Formula I, wherein the pharmaceutically acceptable salt is selected from succinate, mandelate, oxalate, phosphate, hydrochloride, maleate, fumarate, hydrobromide, citrate, sulfate, methanesulfonate, hippurate, benzoate, malate, and acetate; preferably succinate and mandelate; more preferably succinate and D-mandelate.

[0017]

[0018] In some embodiments, the chemical ratio of the compound represented by Formula I to the acid molecule or acid radical is 2:0.5 to 1:3; preferably, the chemical ratio of the compound represented by Formula I to the acid molecule or acid radical is selected from 2:1 to 1:3; more preferably, the chemical ratio of the compound represented by Formula I to the acid molecule or acid radical is selected from 1:0.5, 1:1, 1:1.5 or 1:2.

[0019] This disclosure also provides a succinate A crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 6.170, 11.215, 12.380, 15.558, 18.230, and 18.884.

[0020] In some embodiments, the succinate A crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 6.170, 9.506, 11.215, 12.380, 15.558, 18.230, 18.884, 19.429, 21.392, and 23.609.

[0021] In some embodiments, the X-ray powder diffraction pattern of the succinate A crystal form of the compound shown in Formula I, expressed in terms of the diffraction angle 2θ, is as follows: Figure 2 As shown.

[0022] In some embodiments, the succinate A crystal form of the compound shown in Formula I is wherein the chemical ratio of the compound of Formula I to succinate or succinic acid is about 1:1 to 1:2, preferably 1:1 or 1:2.

[0023] In some embodiments, the succinate A crystal form of the compound shown in Formula I has an endothermic peak value of 97.22 ± 3 °C as shown in the DSC spectrum.

[0024] This disclosure also provides a succinate B crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 6.402, 9.400, 15.728, 17.957, and 19.070.

[0025] In some embodiments, the succinate B crystal form of the compound shown in Formula I has characteristic peaks at 6.402, 9.400, 15.728, 17.957, 19.070, and 21.589.

[0026] In some embodiments, the X-ray powder diffraction pattern of the succinate B crystal form of the compound shown in Formula I, expressed in terms of the diffraction angle 2θ, is as follows: Figure 3 As shown.

[0027] In some embodiments, the succinate B crystal form of the compound shown in Formula I has an endothermic peak value of 121.36 ± 3 °C as shown in the DSC spectrum.

[0028] In some embodiments, the compound represented by Formula I is in succinate B crystal form, wherein the chemical ratio of the compound of Formula I to succinate or succinic acid is about 1:1 to 1:2, preferably 1:1 or 1:2.

[0029] This disclosure also provides a succinate C crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 5.555, 5.588, 9.363, 11.165, 18.229, and 18.668.

[0030] In some embodiments, the succinate C crystal form of the compound shown in Formula I has characteristic peaks at 5.555, 5.588, 9.363, 11.165, 15.505, 18.229, 18.668, 19.734, and 21.920.

[0031] In some embodiments, the succinate C crystal form of the compound shown in Formula I has characteristic peaks at 5.555, 5.588, 9.363, 11.165, 15.505, 18.229, 18.668, 19.734, 21.920, and 26.943.

[0032] In some embodiments, the X-ray powder diffraction pattern of the succinate C crystal form of the compound shown in Formula I, expressed as a diffraction angle 2θ, is as follows: Figure 4 As shown.

[0033] In some embodiments, the succinate C crystal form of the compound shown in Formula I has an endothermic peak value of 93.71±3℃ and 118.69±3℃ as the DSC spectrum.

[0034] In some embodiments, the succinate C crystal form of the compound shown in Formula I is wherein the chemical ratio of the compound of Formula I to succinate or succinic acid is about 1:1 to 1:2, preferably 1:1 or 1:2.

[0035] This disclosure provides a method for preparing the above-mentioned succinate C crystal form, which includes: mixing the compound shown in Formula I with tetrahydrofuran and succinic acid, crystallizing, and obtaining succinate C crystal form.

[0036] This disclosure also provides a succinate D crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, has characteristic peaks at 6.878, 10.856, 11.784, 18.225, and 18.597.

[0037] In some embodiments, the succinate D crystal form of the compound shown in Formula I has characteristic peaks at 6.878, 10.856, 11.784, 15.639, 17.314, 18.225, 18.597, 19.103, 19.755, 20.155, and 21.477.

[0038] In some embodiments, the succinate D crystal form of the compound shown in Formula I has characteristic peaks at 6.878, 10.856, 11.784, 15.639, 17.314, 18.225, 18.597, 19.103, 19.755, 20.155, 21.477, and 27.367.

[0039] In some embodiments, the succinate D crystal form of the compound shown in Formula I has characteristic peaks at 6.878, 10.856, 11.784, 15.639, 17.314, 18.225, 18.597, 19.103, 19.755, 20.155, 21.477, 22.268, 24.672, 25.655, 27.367, 28.023, and 28.715.

[0040] In some embodiments, the X-ray powder diffraction pattern of the succinate D crystal form of the compound shown in Formula I, expressed in terms of the diffraction angle 2θ, is as follows: Figure 5 As shown.

[0041] In some embodiments, the succinate D crystal form of the compound shown in Formula I has an endothermic peak value of 132.93 ± 3 °C as shown in the DSC spectrum.

[0042] In some embodiments, the succinate D crystal form of the compound shown in Formula I is wherein the chemical ratio of the compound of Formula I to succinate or succinic acid is about 1:1 to 1:2, preferably 1:1 or 1:2.

[0043] This disclosure provides a method for preparing the above-mentioned succinate D crystal form, which includes: mixing the compound shown in Formula I with acetone and succinic acid, and crystallizing at low temperature to obtain the succinate D crystal form.

[0044] This disclosure provides a D-mandelate A crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 10.554, 13.168, 15.728, 17.944, and 19.784.

[0045] In some embodiments, the D-mandelate A crystal form of the compound shown in Formula I has characteristic peaks at 6.159, 8.371, 10.554, 13.168, 15.728, 17.944, 19.784, and 20.381.

[0046] In some embodiments, the D-mandelate A crystal form of the compound shown in Formula I has characteristic peaks at 6.159, 8.371, 10.554, 13.168, 15.728, 17.944, 19.784, 20.381, 21.121, and 21.434.

[0047] More preferably, the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is as follows: Figure 6 As shown.

[0048] In some embodiments, the D-mandelate A crystal form of the compound shown in Formula I has an endothermic peak value of 176.10 ± 3 °C as shown in the DSC spectrum.

[0049] In some embodiments, the compound shown in Formula I is in the D-mandelate A crystal form, wherein the chemical ratio of the compound of Formula I to mandelate or mandelic acid is approximately 1:1.

[0050] This disclosure also provides a method for preparing the D-mandelate A crystal form of the compound shown in Formula I, comprising: mixing the compound shown in Formula I with a solvent and D-mandelic acid, and crystallizing to obtain the D-mandelate A crystal form; wherein the solvent is selected from one or more of isopropanol, ethanol, and ethyl acetate, preferably the solvent is selected from isopropanol / ethyl acetate, ethanol / ethyl acetate, and ethyl acetate.

[0051] This disclosure provides an oxalate I crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 4.885, 11.219, 12.092, 14.565, 16.383, and 18.201.

[0052] More preferably, the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is as follows: Figure 7 As shown.

[0053] In some embodiments, the oxalate I crystal form of the compound represented by Formula I has an endothermic peak value of 136.67 ± 3 °C as shown in the DSC spectrum.

[0054] In some embodiments, the compound represented by Formula I is in oxalate form I, wherein the chemical ratio of the compound of Formula I to oxalate or oxalic acid is about 1:1 to 1:3, preferably 1:1 or 1:2.

[0055] This disclosure also provides a method for preparing the oxalate I crystal form of the compound shown in Formula I, which includes: mixing the compound shown in Formula I with acetonitrile and oxalic acid, and crystallizing to obtain the oxalate I crystal form.

[0056] This disclosure provides a hydrochloride I crystal form of the compound shown in Formula I, wherein the X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, has characteristic peaks at 4.891, 11.255, 16.565, 17.947, and 22.056.

[0057] More preferably, the X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is as follows: Figure 9 As shown.

[0058] In some embodiments, the hydrochloride I crystal form of the compound shown in Formula I has a DSC spectrum showing an endothermic peak value of 194.65 ± 3 °C.

[0059] In some embodiments, the hydrochloride form I of the compound represented by Formula I is used, wherein the chemical ratio of the compound of Formula I to chloride ions or hydrogen chloride is about 2:1 to 1:3, preferably 1:1 or 1:2.

[0060] This disclosure also provides a method for preparing hydrochloric acid of the compound shown in Formula I, which includes: mixing the compound shown in Formula I with acetonitrile and hydrochloric acid, and crystallizing to obtain hydrochloride salt I crystal form.

[0061] This disclosure also provides an amorphous form of the phosphate of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in Figure 1. Figure 8 As shown.

[0062] This disclosure also provides an amorphous form of the maleate salt of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in the figure. Figure 10 As shown.

[0063] This disclosure also provides an amorphous form of the fumarate of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in the figure. Figure 11 As shown.

[0064] This disclosure also provides an amorphous form of the hydrobromide of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in Figure 1. Figure 12 As shown.

[0065] This disclosure also provides an amorphous form of the citrate salt of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably, its X-ray powder diffraction pattern is as follows: Figure 13 As shown.

[0066] This disclosure also provides an amorphous form of the sulfate of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in the figure. Figure 14 As shown.

[0067] This disclosure also provides an amorphous form of the methanesulfonate of Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in Figure 1. Figure 15 As shown.

[0068] This disclosure also provides an amorphous form of hippurate, a compound of formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in the figure. Figure 16 As shown.

[0069] This disclosure also provides an amorphous form of the benzoate of Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in Figure 1. Figure 17 As shown.

[0070] This disclosure also provides an amorphous form of the malate of Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in Figure 1. Figure 18 As shown.

[0071] This disclosure also provides an amorphous form of the acetate of the compound shown in Formula I, whose X-ray powder diffraction pattern has no sharp peaks, preferably has non-sharp peaks, and more preferably has an X-ray powder diffraction pattern as shown in the figure. Figure 19 As shown.

[0072] The crystallization methods described in this disclosure are conventional, such as volatilization crystallization, cooling crystallization, pulping crystallization, room temperature crystallization, and stirring crystallization.

[0073] This disclosure also provides a pharmaceutical composition comprising the above-described amorphous material, a pharmaceutically acceptable salt, a crystalline form or a crystalline form prepared by the above-described method, and optionally a pharmaceutically acceptable excipient.

[0074] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the amorphous substance, pharmaceutically acceptable salt, crystal form or crystal form obtained by the above preparation method with a pharmaceutically acceptable excipient.

[0075] This disclosure also provides the use of the above-described amorphous material, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-described preparation method, or the above-described pharmaceutical composition in the preparation of a medicament for regulating the ubiquitination and degradation of androgen receptor (AR) proteins.

[0076] This disclosure also provides the use of the above-mentioned amorphous material, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-mentioned method, or the above-mentioned pharmaceutical composition for regulating the ubiquitination and degradation of androgen receptor (AR) proteins.

[0077] This disclosure also provides the above-mentioned amorphous substance, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-mentioned preparation method, or the above-mentioned pharmaceutical composition for regulating the ubiquitination and degradation of androgen receptor (AR) protein.

[0078] This disclosure also provides the use of the above-described amorphous material, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-described preparation method, or the above-described pharmaceutical composition in the preparation of a medicament for treating and / or preventing androgen receptor-mediated or dependent diseases or conditions; wherein androgen receptor-mediated or dependent diseases or conditions are preferably selected from tumors, male sexual dysfunction, and Kennedy's disease.

[0079] This disclosure also provides the use of the above-mentioned amorphous material, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-mentioned preparation method, or the above-mentioned pharmaceutical composition for the treatment and / or prevention of androgen receptor-mediated or dependent diseases or conditions; wherein the androgen receptor-mediated or dependent diseases or conditions are preferably selected from tumors, male sexual dysfunction, and Kennedy's disease.

[0080] This disclosure also provides the above-described amorphous substance, pharmaceutically acceptable salt, crystal form, crystal form prepared by the above-described preparation method, or pharmaceutical composition for treating and / or preventing androgen receptor-mediated or dependent diseases or conditions, wherein the androgen receptor-mediated or dependent diseases or conditions are preferably selected from tumors, male sexual dysfunction, and Kennedy's disease.

[0081] In some implementations, the androgen receptor-mediated or dependent disease or condition is selected from prostate cancer, benign prostatic hyperplasia, hirsutism, alopecia, anorexia nervosa, breast cancer, acne, male sexual dysfunction, Kennedy's disease, and AIDS, preferably prostate cancer, more preferably hormone-sensitive prostate cancer or hormone-refractory prostate cancer.

[0082] 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 ±0.2 (including the case where the number has more than one decimal place after rounding), 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.

[0083] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2015 edition of the Chinese Pharmacopoeia,

[0084] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0085] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0086] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0087] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0088] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0089] The “differential scanning calorimetry or DSC” described in this disclosure refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0090] The drying temperature described in this disclosure is generally 25℃~100℃, preferably 40℃~70℃. It can be dried under normal pressure or reduced pressure, with a pressure <-0.08MPa.

[0091] The “excipients” described in this disclosure include, but are not limited to, any adjuvants, carriers, flow aids, sweeteners, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, or emulsifiers that have been approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0092] The "pulping" described in this disclosure refers to a purification method that utilizes the characteristic that substances have poor solubility in solvents, but impurities have good solubility in solvents. Pulping purification can remove color, change crystal form, or remove a small amount of impurities.

[0093] The starting material used in the crystal form preparation method disclosed herein can be any form of compound, including but not limited to: amorphous substances, arbitrary crystal forms, hydrates, solvates, etc.

[0094] The numerical values ​​in this disclosure, such as those relating to the content of certain substances, are calculated data and inevitably contain a certain degree of error. Generally, ±10% is within the reasonable error range. The error may vary to some extent depending on the context in which it is used, but this variation shall not exceed ±10%, and may be ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, or ±1%, preferably ±5%.

[0095] Compound I of this disclosure, 2-chloro-4-((S)-8-(4-(4-((4-(3-((((S)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile (hereinafter referred to as Compound I), was prepared according to the method in PCT / CN2022 / 134333, and the relevant content is cited in this text for illustration. Attached Figure Description

[0096] Figure 1 The image shows the XPRD spectrum of the amorphous material of Formula I.

[0097] Figure 2 The image shows the XPRD spectrum of succinate A of Formula I.

[0098] Figure 3 The image shows the XPRD spectrum of succinate B crystal form of Formula I.

[0099] Figure 4 The image shows the XPRD spectrum of the C crystal form of succinate of formula I.

[0100] Figure 5 The image shows the XPRD spectrum of the D crystal form of succinate of Formula I.

[0101] Figure 6 The image shows the XPRD spectrum of the A crystal form of formula I mandelate.

[0102] Figure 7 The image shows the XPRD spectrum of oxalate I crystal form of Formula I.

[0103] Figure 8 The image shows the XPRD spectrum of the amorphous phosphate of formula I.

[0104] Figure 9 The image shows the XPRD spectrum of the I-type hydrochloride.

[0105] Figure 10 The image shows the XPRD spectrum of the amorphous maleate of formula I.

[0106] Figure 11 The image shows the XPRD spectrum of the amorphous fumarate of Formula I.

[0107] Figure 12 The image shows the XPRD spectrum of the amorphous form of hydrobromide of formula I.

[0108] Figure 13 The image shows the XPRD spectrum of the amorphous citrate of formula I.

[0109] Figure 14 The image shows the XPRD spectrum of the amorphous citrate of formula I.

[0110] Figure 15 The image shows the XPRD spectrum of the amorphous form of methanesulfonate of formula I.

[0111] Figure 16 The image shows the XPRD spectrum of the amorphous form of hippurate of formula I.

[0112] Figure 17 The image shows the XPRD spectrum of the amorphous form of benzoate of formula I.

[0113] Figure 18 The image shows the XPRD spectrum of the amorphous malate of formula I.

[0114] Figure 19 The image shows the XPRD spectrum of the amorphous acetate of formula I. Detailed Implementation

[0115] The following embodiments are used to further describe this disclosure, but these embodiments are not intended to limit the scope of this disclosure.

[0116] Example

[0117] 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).

[0118] MS measurements were performed using an Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS liquid chromatography-mass spectrometry system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS).

[0119] waters ACQuity UPLC-QD / SQD (Manufacturer: waters, MS model: waters ACQuity QdaDetector / waters SQ Detector)

[0120] THERMO Ultimate 3000-Q Exactive (Manufacturer: THERMO, MS Model: THERMOQExactive)

[0121] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLC e2695-2489 HPLC system.

[0122] Chiral HPLC analysis was performed using an Agilent 1260DAD high-performance liquid chromatograph.

[0123] High performance liquid chromatography (HPLC) was performed using Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson GX-281 preparative chromatographs.

[0124] Chiral preparation was performed using a Shimadzu LC-20AP preparative chromatograph.

[0125] The CombiFlash rapid preparation system uses a CombiFlash Rf200 (TELEDYNE ISCO).

[0126] Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF254 or Qingdao GF254. The silica gel plates used in thin-layer chromatography (TLC) have a diameter of 0.15 mm to 0.2 mm, and the diameter of the silica gel plates used for thin-layer chromatography separation and purification products is 0.4 mm to 0.5 mm.

[0127] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0128] Mean inhibition rate of kinases and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG GmbH, Germany).

[0129] The known starting materials disclosed herein can be synthesized using or in accordance with methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, and Darui Chemicals.

[0130] Unless otherwise specified in the examples, all reactions can be carried out under an argon or nitrogen atmosphere.

[0131] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0132] A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon with a volume of approximately 1L.

[0133] The pressurized hydrogenation reaction was performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0134] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0135] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0136] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0137] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0138] 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.

[0139] XRPD (X-ray Powder Diffraction) was used for analysis: measurements were performed using a BRUKER D8 X-ray diffractometer. Specific data collected included: Cu anode (40 kV, 40 mA), Cu-Kα1 rays. Kα2 rays Kβ rays Scan range (2θ range): 3~45°.

[0140] 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.

[0141] 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 30-350℃). The nitrogen purging rate was 50mL / min.

[0142] DVS stands for Dynamic Moisture Adsorption: The detection method is SMSDVS Advantage. At 25℃, the humidity changes from 50% to 95% to 0% to 95% to 50%, with a step size of 10% (the final step is 5%). (The specific humidity range is subject to the corresponding spectrum. The methods listed here are the most commonly used methods.) The judgment criteria are Tmax ≤ 360 min and dm / dt not greater than 0.002%.

[0143] Ion chromatography detection: Instrument: DIONX AQUION ion chromatograph (USA); Detection method: conductivity; Separation column: IonPac AS14A-AG14; Eluent: NaHCO3 0.5 mM + Na2CO3 4.0 mM; Flow rate: 1.0 mL / min. Example 1: The compound shown in Formula I was synthesized using the preparation method of Example 15-1 in patent application PCT / CN2022 / 134333.

[0144] 2-Chloro-4-((S)-8-(4-(4-((4-(3-((((S)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile 1-1(Formula I)

[0145] 2-Chloro-4-((S)-8-(4-(4-((4-(3-((((R)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile 1-2

[0146]

[0147] first step

[0148] (±)-4-((4-(3-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester 1b

[0149] Add 3 mL of a mixture of dichloromethane and methanol (V / V = 2 / 1) to a 25 mL three-necked flask, then add (±)-3-((3-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione 1a (90 mg, 0.25 mmol, prepared by the method disclosed in Compound 86 on page 266 of patent application "WO2018237026A1"), anhydrous sodium acetate (102 mg, 1.24 mmol), react for 15 minutes, then add 4-formylpiperidine-1-carboxylic acid tert-butyl ester (85 mg, 0.40 mmol), react for 30 minutes, then slowly add sodium triacetoxyborohydride (106 mg, 0.50 mmol), and react for 16 hours. The reaction was quenched by adding 10 mL of saturated sodium bicarbonate solution to the system under ice bath cooling. The mixture was extracted with dichloromethane (20 mL × 3), and the organic phases were combined and washed with saturated sodium chloride solution (20 mL × 3). The organic phases were dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system B to give title compound 1b (110 mg, yield: 90%).

[0150] MS m / z(ESI):486.6[M+1].

[0151] Step 2

[0152] (±)-3-((3-(4-(piperidin-4-ylmethyl)piperazin-1-yl)phenyl)amino)piperidin-2,6-dione dihydrochloride 1c

[0153] Compound 1b (105 mg, 0.22 mmol) was dissolved in 3 mL of dichloromethane. Under ice bath conditions, a 0.53 mL solution of 4 M hydrogen chloride in 1,4-dioxane was slowly added, and the reaction was allowed to proceed for 4 hours. The reaction mixture was concentrated and dried under vacuum to obtain the crude product, title compound 1c (99 mg, yield: 98%). The crude product was used directly in the next reaction without purification.

[0154] MS m / z(ESI): 386.5 [M+1].

[0155] Step 3

[0156] 2-Chloro-4-((3S)-8-(4-(4-((4-(3-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile 1 (a mixture of diastereomers)

[0157] 2 mL of N,N-dimethylformamide, compound 1c (50 mg, 0.094 mmol), (S)-4-(2-(3-chloro-4-cyanophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-8-yl)-3-fluorobenzoic acid 1d (45 mg, 0.083 mmol, prepared by the method disclosed in Example 52 on page 271 of patent application “WO2021055756A1”) were added to a 25 mL three-necked flask in sequence. N,N-diisopropylethylamine (85 mg, 0.66 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (63 mg, 0.16 mmol) were added slowly under ice bath. The cold bath was removed and the reaction was allowed to proceed for 16 hours. The reaction solution was filtered and prepared by high performance liquid chromatography (Waters 2545, elution system: 10 mmol / L ammonium bicarbonate aqueous solution and acetonitrile, acetonitrile gradient: 50%-95%, flow rate: 30 mL / min) to give title compound 1 (a mixture of diastereomers, ratio 1:1, 36 mg, yield: 54%).

[0158] MS m / z(ESI): 795.3 [M+1];

[0159] 1H NMR (500MHz, DMSO-d6): δ10.75(s,1H),7.59(d,1H),7.13(d,1H),7.11(s,1H),7.06(t,1H),6.89(t,1H),6 .79(d,1H),6.65(dd,1H),6.24(s,1H),6.17(d,1H),6.12(d,1H),5.60(d,1H),4.35-4.25(m,1H),4.07-3.9 7(m,1H),3.43(d,1H),3.34-3.30(m,3H),3.17-2.92(m,8H),2.78-2.68(m,1H),2.64-2.50(m,3H),2.48-2 .40(m,4H),2.28-1.93(m,4H),1.90-1.67(m,5H),1.62-1.48(m,3H),1.27-1.15(m,4H),1.12-0.99(m,2H).

[0160] Step 4

[0161] 2-Chloro-4-((S)-8-(4-(4-((4-(3-((((S)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile 1-1(Formula I)

[0162] 2-Chloro-4-((S)-8-(4-(4-((4-(3-((((R)-2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-carbonyl)-2-fluorophenyl)-3-methyl-2,8-diazaspiro[4.5]dec-2-yl)benzonitrile 1-2

[0163] Compound 1 (21 mg) was chirally separated (separation conditions: Gilson-281, column: Phenomenex Amylose-2, 5 μm, 21.2 mm * 250 mm, mobile phase: acetonitrile (0.5% 7M ammonia (NH3) in methanol) / ethanol (0.5% 7M ammonia (NH3) in methanol) = 40 / 60 (v / v)), flow rate 20 mL / min), yielding title compound 1-2 (5 mg) and compound 1-1 (8 mg).

[0164] 1-2 (5mg, yield: 24%), shorter retention time:

[0165] MS m / z(ESI): 795.3 [M+1].

[0166] Chiral HPLC analysis method: retention time 3.53 min (Agilent 1260 DAD, column: Phenomenex LUX Amylose-2, (4.6 × 150 mM), 5 μm; mobile phase: acetonitrile / ethanol / diethylamine = 40 / 60 / 0.06 (v / v / v), flow rate 1 mL / min);

[0167] 1 H NMR (500MHz, DMSO-d6): δ10.75(s,1H),7.59(d,1H),7.14(d,1H),7.12(s,1H),7.06(t,1H),6.90(t,1H),6 .79(d,1H),6.65(dd,1H),6.24(s,1H),6.17(d,1H),6.13(d,1H),5.61(d,1H),4.35-4.25(m,1H),4.07-3.9 7(m,1H),3.43(d,1H),3.37-3.30(m,3H),3.17-2.92(m,8H),2.78-2.68(m,1H),2.64-2.50(m,3H),2.48-2 .40(m,4H),2.28-1.93(m,4H),1.90-1.67(m,5H),1.62-1.48(m,3H),1.27-1.15(m,4H),1.12-0.99(m,2H).

[0168] Compound 1-1 (8 mg, yield: 38%), longer retention time:

[0169] MS m / z(ESI): 795.3 [M+1].

[0170] Chiral HPLC analysis method: retention time 4.19 min (Agilent 1260 DAD, column: Phenomenex LUX Amylose-2, (4.6 × 150 mM), 5 μm; mobile phase: acetonitrile / ethanol / diethylamine = 40 / 60 / 0.06 (v / v / v), flow rate 1 mL / min);

[0171] 1H NMR (500MHz, DMSO-d6): δ10.76(s,1H),7.59(d,1H),7.13(d,1H),7.11(s,1H),7.06(t,1H),6.90(t,1H),6 .80(d,1H),6.65(dd,1H),6.24(s,1H),6.17(d,1H),6.12(d,1H),5.61(d,1H),4.35-4.25(m,1H),4.07-3.9 7(m,1H),3.43(d,1H),3.34-3.30(m,3H),3.17-2.92(m,8H),2.78-2.68(m,1H),2.64-2.50(m,3H),2.48-2 .40(m,4H),2.28-1.93(m,4H),1.90-1.67(m,5H),1.62-1.48(m,3H),1.27-1.15(m,4H),1.12-0.99(m,2H).

[0172] Biological evaluation

[0173] The following test examples further describe and explain this disclosure, but these test examples are not intended to limit the scope of this disclosure.

[0174] Test Example 1: Determination of the degradation activity of the disclosed compound on androgen receptors in LNCaP cells.

[0175] The in vitro cell experiments described below can determine the degradation activity of the test compound on the androgen receptor (AR) in human prostate cancer cell lines. This activity can be measured using DCs. 50Values ​​are used to represent the results. On the first day of the experiment, LNCaP cells (ATCC, CRL-1740) were seeded at a density of 20,000 cells / well in 96-well plates using phenol red-free RPMI 1640 medium (Gibco, 11835-030) containing 5% activated charcoal-treated fetal bovine serum (Shanghai Bosheng Biotechnology Co., Ltd., S-FBS-AU-045), with 200 μL of cell suspension per well. The plates were incubated at 37°C in a 5% CO2 cell culture incubator for three days. On the fourth day, 22 μL of the test compound prepared with a serially diluted medium containing 1 nM R1881 (Aladdin, M305037) was added to each well. The final concentration of R1881 was 0.1 nM. The final concentration of the compound was determined by a 6-fold serial dilution starting from 2.5 μM. A blank control cell well containing 0.5% DMSO was also included. The plates were incubated at 37°C in a 5% CO2 cell culture incubator for 24 hours. On day 5, remove the 96-well cell culture plate, aspirate and discard the cell culture supernatant, and wash the cells once with 300 μL of ice-cold PBS. Then add 50 μL of 1× lysis buffer (Cell Signaling Technology, #9803S) diluted with ddH2O and containing 1 mM PMSF. Vortex for 1 minute, then place the plate on ice for 15–20 minutes for lysis. After mixing by pipetting, centrifuge at 4000 rpm, 4°C for 10 minutes. Use the AR ELISA kit (Cell Signaling Technology, #12850) to detect AR protein levels according to the manufacturer's instructions. Add 96 μL of sample diluent and 4 μL of sample lysis buffer or blank lysis buffer to each well of the 96-well plate. Seal the plate, vortex to mix, and incubate at 37°C for 2 hours. Discard the liquid in the wells, wash the plate five times with 300 μL of washing buffer per well, add 100 μL of working antibody solution, seal the plate, and incubate at 37°C for 1 hour. Discard the liquid in each well. Wash the plate five times with 300 μL of washing buffer, then add 100 μL of enzyme conjugate working solution. Seal the plate and incubate at 37°C for 30 minutes. Discard the liquid in each well. Wash the plate five times with 300 μL of washing buffer, then add 100 μL of chromogenic solution. Seal the plate and incubate at room temperature in the dark for 5 minutes. Add 100 μL of stop solution to each well. After mixing, immediately place the plate on a microplate reader (BMG Labtech, PHERAstar FS) to read the OD450 and OD540 values. Subtract the corresponding OD540 absorbance value from the OD450 absorbance values ​​of all wells to obtain the corrected OD450 absorbance value (OD450-correction). Calculate the degradation rate of each compound concentration using the following formula, where the maximum value (Max) is the maximum degradation rate of the compound's AR. Use GraphPad Prism to perform curve fitting based on the logarithmic concentration and degradation efficiency of the compound and calculate the DC. 50 value.

[0176] Degradation rate (%) = (OD450 - calibration) 空白对照 -OD450-calibration 化合物 ) / (OD450-calibration 空白对照 -OD450-calibration 裂解液对照 )×100%

[0177] The bioactivity of the compound disclosed herein is obtained from the above analysis, and the calculated DC is... 50 The maximum values ​​are shown in Table 1 below.

[0178] Table 1. Degradation activity of the disclosed compounds on androgen receptors in LNCaP cells.

[0179]

[0180] Conclusion: The compound disclosed in this study exhibits significant degradation activity against androgen receptors in LNCaP cells.

[0181] Test Example 2: Determination of the inhibitory effect of the disclosed compound on the proliferation of LNCaP

[0182] The in vitro cell experiments described below can determine the inhibitory activity of the test compound on the proliferation of human prostate cancer cell lines. This activity can be measured using the IC50 assay. 50 Values ​​are used to represent the results. On the first day of the experiment, LNCaP cells (ATCC, CRL-1740) were seeded at a density of 7500 cells / well in 96-well plates using phenol red-free RPMI 1640 medium (Gibco, 11835-030) containing 5% activated charcoal-treated fetal bovine serum (Shanghai Bosheng Biotechnology Co., Ltd., S-FBS-AU-045), with 200 μL of cell suspension per well. The plates were then incubated at 37°C in a 5% CO2 cell culture incubator for three days. On day four, 22 μL of the serially diluted test compound prepared with plating medium containing 1 nM R1881 (Aladdin, M305037) was added to each well. The final concentration of R1881 was 0.1 nM. The final concentration of the compound was determined by nine 4-fold serial dilutions starting from 10 μM. Blank control wells containing 0.5% DMSO and cell-free solvent control wells were included. The plate was incubated at 37°C in a 5% CO2 incubator for six days. On day ten, the 96-well cell culture plate was removed, and 110 μL of cell culture supernatant was aspirated from each well and discarded. Then, 50 μL of the supernatant was added to each well. The Luminescent CellViability Assay (Promega, G7573) was incubated at room temperature for 10 minutes, and the luminescence signal value (RLU) was read using a PerkinElmer, VICTOR 3 microplate reader. The inhibition rate of each compound concentration was calculated using the following formula, where the maximum value (Max) is the maximum inhibition rate of the compound against cell proliferation. The IC50 was calculated by curve fitting based on the logarithmic concentration and inhibition rate of the compound using GraphPad Prism. 50 value.

[0183] Inhibition rate (%) = (RLU) 空白对照 -RLU 化合物 ) / (RLU 空白对照 -RLU 溶媒对照 )×100%

[0184] The bioactivity of the compound disclosed herein is obtained from the above analysis, and the calculated IC50 value is... 50 The values ​​are shown in Table 2 below.

[0185] Table 2. Inhibitory activity of the disclosed compounds against the proliferation of LNCaP cells.

[0186]

[0187] Conclusion: The compound disclosed herein exhibits good inhibitory activity against the proliferation of LNCaP cells.

[0188] Test Example 3: Pharmacokinetic Evaluation

[0189] Comparative Example 1 (synthesized according to Patent WO2021055756A1, Example 307), has the following structure:

[0190]

[0191] ARV-110 (synthesized according to patent WO2018071606A1, Example 406) has the following structure:

[0192]

[0193] I. SD Rat Experiment

[0194] 1. Abstract

[0195] Using SD rats as test animals, the plasma drug concentration of the disclosed compound at different time points after gavage (ig) administration to SD rats was determined by LC / MS / MS. The pharmacokinetic behavior of the disclosed compound in SD rats was investigated to evaluate its pharmacokinetic characteristics.

[0196] 2. Test Plan

[0197] 2.1 Test Drugs

[0198] Compound of Example 1;

[0199] Compounds from Example 1-1;

[0200] Comparative Example 1 compound.

[0201] 2.2 Experimental Animals

[0202] Twelve SD rats, half male and half female, were randomly divided into three groups and provided by Vital River Laboratory Animal Co., Ltd. After fasting overnight, the rats were administered the drugs by gavage.

[0203] 2.3 Drug Preparation

[0204] Weigh a certain amount of the compound to be tested, add 5% DMSO + 5% Tween 80 + 90% physiological saline, and prepare a clear solution of 0.2 mg / mL.

[0205] 2.4 Administration

[0206] The dosage is 2.0 mg / kg, and the administration volume is 10.0 mL / kg.

[0207] 3. Operation

[0208] Blood samples of 0.2 mL were collected from the orbital cavity before administration and at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, and 24.0 hours after administration. The samples were placed in EDTA-K2 anticoagulant tubes, centrifuged at 10,000 rpm for 1 minute (4°C), and plasma was separated within 1 hour and stored at -20°C for analysis. The entire process, from blood collection to centrifugation, was performed under ice bath conditions. Patients ate 2 hours after administration.

[0209] To determine the content of the target compound in the plasma of SD rats after administration of different drug concentrations: 25 μL of plasma samples from SD rats at each time point after drug administration were taken, 200 μL of acetonitrile was added to precipitate the protein and 50 μL of camptothecin (100 ng / mL), the mixture was vortexed for 5 minutes and centrifuged at 4000 rpm for 10 minutes. 30 μL of the supernatant was taken, diluted with 120 μL of water, and 1 μL was taken for LC / MS / MS analysis.

[0210] 4. Pharmacokinetic Parameter Results

[0211] Table 3. Pharmacokinetic parameters of the disclosed compounds in SD rats.

[0212]

[0213] Conclusion: Compared with Comparative Example 1, the compound disclosed herein has a larger area under the curve, a longer half-life, and a lower clearance rate in SD rats, exhibiting excellent pharmacokinetic activity and significant pharmacokinetic advantages.

[0214] II. Dog Testing

[0215] 1. Abstract

[0216] Using beagle dogs as test animals, the plasma drug concentrations at different time points after gavage (ig) administration of the compound disclosed herein were determined by LC / MS / MS. The pharmacokinetic behavior of the compound disclosed herein in beagle dogs was investigated to evaluate its pharmacokinetic characteristics.

[0217] 2. Test Plan

[0218] 2.1 Test Drugs

[0219] Compounds from Example 1-1;

[0220] Compound ARV-110.

[0221] 2.2 Experimental Animals

[0222] Eight Beagle dogs, half male and half female, were divided into two groups and provided by Jiangsu Zhaoshengyuan Biotechnology Co., Ltd. After fasting overnight, they were administered the medication by gavage.

[0223] 2.3 Drug Preparation

[0224] Weigh a certain amount of the compound to be tested, add 5% DMSO + 30% PG + 30% PEG400 + 35% physiological saline to prepare a 0.4 mg / mL solution.

[0225] 2.4 Administration

[0226] The dosage is 2.0 mg / kg, and the administration volume is 5.0 mL / kg.

[0227] 3. Operation

[0228] Blood samples of 0.5 mL were collected from the forelimb vein before administration and at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 11.0, and 24.0 hours after administration. The samples were placed in EDTA-K2 anticoagulant tubes, centrifuged at 10,000 rpm for 2 minutes (4°C), and plasma was separated within 1 hour and stored at -80°C for analysis. The entire process, from blood collection to centrifugation, was performed under ice bath conditions. Patients ate 3 hours after administration.

[0229] To determine the content of the target compound in the plasma of beagle dogs after administration of different concentrations of the drug: 50 μL of plasma samples from beagle dogs at each time point after drug administration were taken, and 25 μL of camptothecin (1 μg / mL) and 450 μL of acetonitrile were added to precipitate the protein. The mixture was vortexed and centrifuged at 3700 rpm for 10 minutes. 0.5 μL of the supernatant was taken for LC / MS / MS analysis.

[0230] 4. Pharmacokinetic Parameter Results

[0231] Table 4. Pharmacokinetic parameters of the disclosed compounds in beagle dogs.

[0232]

[0233] Conclusion: Compared with compound ARV-110, the compound disclosed herein has a larger area under the curve and a lower clearance rate in beagle dogs, exhibiting excellent pharmacokinetic absorption activity and significant pharmacokinetic advantages.

[0234] III. Monkey Experiments

[0235] 1. Abstract

[0236] Using cynomolgus monkeys as test animals, the plasma drug concentrations at different time points after gavage (ig) administration of the compound disclosed herein were determined by LC / MS / MS. The pharmacokinetic behavior of the compound disclosed herein in cynomolgus monkeys was investigated to evaluate its pharmacokinetic characteristics.

[0237] 2. Test Plan

[0238] 2.1 Test Drugs

[0239] Compounds from Example 1-1;

[0240] ARV-110.

[0241] 2.2 Experimental Animals

[0242] Eight cynomolgus macaques, half male and half female, were divided into two groups and provided by Medicilon Medical Technology (Shanghai) Co., Ltd. After fasting overnight, they were administered medication by gavage.

[0243] 2.3 Drug Preparation

[0244] Weigh a certain amount of the compound to be tested, add 5% DMSO + 30% PG + 30% PEG400 + 35% physiological saline, and prepare a clear solution of 0.4 mg / mL.

[0245] 2.4 Administration

[0246] The dosage is 2.0 mg / kg, and the administration volume is 5.0 mL / kg.

[0247] 3. Operation

[0248] Blood samples of 1 mL were collected from the forelimb vein before administration and at 0.25, 0.5, 1.0, 2.0, 4.0, 6.0, 8.0, 12.0, and 24.0 hours after administration. The samples were placed in EDTA-K2 anticoagulant tubes, centrifuged at 2200g for 10 minutes (2-8℃), and the plasma was separated within 1 hour and stored at -80℃ for analysis. The entire process from blood collection to centrifugation was performed under ice bath conditions. Patients were allowed to eat and drink freely 3 hours after administration.

[0249] Determination of the content of the target compound in cynomolgus monkey plasma after administration of different drug concentrations: Take 20 μL of cynomolgus monkey plasma samples at each time point after administration, add 400 μL of methanol to precipitate proteins (containing 10 ng / mL verapamil), vortex mix for 1 minute, centrifuge at 18000g for 7 minutes, and take 5 μL of supernatant for LC / MS / MS analysis.

[0250] 4. Pharmacokinetic Parameter Results

[0251] Table 5 shows the pharmacokinetic parameters of the disclosed compounds in cynomolgus monkeys:

[0252]

[0253] Conclusion: Compared with ARV-110, the compound disclosed herein has a larger area under the curve and a lower clearance rate in cynomolgus monkeys, exhibiting excellent pharmacokinetic absorption activity and significant pharmacokinetic advantages.

[0254] Example 2: Preparation of free amorphous materials

[0255] 10 mg of the compound shown in Formula I was added to 1 mL of solvent A. The sample was stirred and slurried at room temperature, filtered, and the filter cake was collected and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was a free amorphous substance, and the XRPD spectrum is shown below. Figure 1 As shown in the figure. The DSC spectrum shows an endothermic peak at 122.02℃. The TGA spectrum shows a weight loss of 5.06% from 33 to 170℃.

[0256] Table 6

[0257]

[0258] Example 3: Preparation of Free Amorphous Materials

[0259] The compound shown in Formula I was dissolved in solvent B, and after adding an antisolvent, it precipitated out. The precipitate was then obtained by centrifugation and vacuum drying. X-ray powder diffraction analysis showed that the product was amorphous.

[0260] Table 7

[0261]

[0262] Example 4: Preparation of Succinate A Crystal Form

[0263] 10 mg of the compound shown in Formula I was dissolved in 0.3 mL of acetone, followed by the addition of 3.26 mg of solid succinic acid. The mixture was stirred overnight at room temperature, centrifuged, and vacuum dried to obtain the solid. X-ray powder diffraction analysis identified the product as succinate A crystal form. The XRPD spectrum is shown below. Figure 2 As shown in Table 8, the positions of its characteristic peaks are as follows. The DSC spectrum shows an endothermic peak at 97.22℃. The TGA spectrum shows a weight loss of 1.51% from 35-71℃ and a weight loss of 7.23% from 71℃-131℃. Ion chromatography results show a succinate ion content of 14.8%.

[0264] Table 8

[0265]

[0266] Example 5: Preparation of Succinate B Crystal Form

[0267] 10 mg of the compound shown in Formula I was dissolved in 0.1 mL of acetone, followed by the addition of 1.63 mg of solid succinic acid. The mixture was stirred overnight at room temperature to precipitate a solid, which was then centrifuged and dried under vacuum. X-ray powder diffraction analysis identified the product as succinate B crystal form, and the XRPD spectrum is shown below. Figure 3 As shown in Table 9, the positions of its characteristic peaks are as follows. The DSC spectrum shows an endothermic peak at 121.36℃. The TGA spectrum shows a weight loss of 2.19% from 36-120℃. Ion chromatography results show a succinate ion content of 13.2%.

[0268] Table 9

[0269]

[0270] Example 6 Preparation of Succinate C Crystal Form

[0271] 10 mg of the compound shown in Formula I was dissolved in 0.3 mL of tetrahydrofuran, followed by the addition of 3.26 mg of solid succinic acid. The mixture was stirred overnight at room temperature to precipitate a solid, which was then centrifuged and dried under vacuum. X-ray powder diffraction analysis identified the product as succinate C crystal form. The XRPD spectrum is shown below. Figure 4 As shown in Table 10, the positions of its characteristic peaks are as follows. The DSC spectrum shows endothermic peaks at 93.71℃ and 118.69℃. The TGA spectrum shows a weight loss of 8.96% from 31 to 131℃. Ion chromatography results show a succinate ion content of 19.3%.

[0272] Table 10

[0273]

[0274] Example 7 Preparation of Succinate D Crystal Form

[0275] 10 mg of the compound shown in Formula I was dissolved in 0.3 mL of acetone, followed by the addition of 1.63 mg of solid succinic acid. The mixture was stirred overnight at room temperature without precipitation. The mixture was then stirred overnight at 5°C until a solid precipitated. The solid was centrifuged and dried under vacuum. X-ray powder diffraction analysis identified the product as succinate D crystal form. The XRPD spectrum is shown below. Figure 5 As shown in Table 11, the positions of its characteristic peaks are as follows. The DSC spectrum shows an endothermic peak at 132.93℃. The TGA spectrum shows a weight loss of 1.63% at 36-80℃ and 4.00% at 80-140℃. Ion chromatography results show a succinate ion content of 12.9%.

[0276] Table 11

[0277]

[0278] Example 8: Preparation of D-mandelate A crystal form

[0279] The compound shown in Formula I (100 mg, 125.72 mmol) was dissolved in 8 mL of a mixed solvent of isopropanol and ethyl acetate (V / V = 1 / 1), and 0.5 mL of an ethyl acetate solution of D-mandelic acid (19.13 mg, 125.72 μmol) was added. The mixture was stirred at room temperature for 48 hours. A solid precipitated, the reaction mixture was filtered, the filter cake was collected, and dried under vacuum to give the title product (100 g, yield: 84.7%).

[0280] 1 1H NMR characterization: The compound shown in Formula I forms a salt with D-mandelic acid in a 1:1 ratio.

[0281] 1H NMR(500MHz,DMSO)δ10.76(d,1H),7.64-7.57(m,1H),7.41(t,2H),7.38-7.31(m,2H),7.28(q,1H),7.18-7.11(m,2H),7. 08(d,1H),6.91(s,1H),6.81(d,1H),6.70-6.64(m,1H),6.26(s,1H),6.17(dt,2H),5.62(t,1H),5.02-4.98(m,1H),4.31 (t,1H),4.04(q,1H),3.45(d,1H),3.39-3.34(m,3H),3.17-3.06(m,8H),3.02(s,1H),2.79-2.70(m,1H),2.58-2.54(m,3 H),2.30-2.19(m,3H),2.10(s,1H),1.94-1.85(m,2H),1.79(s,2H),1.74(s,2H),1.58(dd,3H),1.22(q,4H),1.09(p,2H).

[0282] X-ray powder diffraction analysis revealed its 2θ as shown in Table 12, defining it as D-mandelate A crystal form. The X-ray powder diffraction pattern is shown below. Figure 6 As shown.

[0283] The DSC spectrum shows that the endothermic peak has a peak value of 176.10℃.

[0284] The TGA spectrum shows a weight loss of 0.91% between 40℃ and 100℃.

[0285] DVS experimental data showed that under normal storage conditions (25℃, 60% RH), the sample's moisture absorption weight gain was approximately 0.833%; under accelerated testing conditions (70% RH), the moisture absorption weight gain was approximately 1.047%; and under extreme conditions (90% RH), the moisture absorption weight gain was approximately 2.245%. During humidity changes from 0% to 95% RH, the desorption and adsorption processes of the sample overlapped. XRPD spectra showed that the crystal form remained unchanged before and after DVS testing.

[0286] Table 12

[0287]

[0288] Example 9: Preparation of D-mandelate A crystal form

[0289] The compound shown in Formula I (1.1 g, 1.38 mmol) was dissolved in 40 mL of ethyl acetate, and 5 mL of an ethyl acetate solution of D-mandelic acid (231.46 mg, 1.52 mmol) was added. The reaction solution was dissolved completely, and the mixture was stirred at room temperature for 48 hours. A solid precipitated, the reaction solution was filtered, the filter cake was collected, and dried under vacuum to give the title product (1.2 g, yield: 92.3%).

[0290] 1 1H NMR characterization: The compound shown in Formula I forms a salt with D-mandelic acid in a 1:1 ratio.

[0291] H NMR(500MHz,DMSO)δ10.76(s,1H),7.60(d,1H),7.41(d,2H),7.34(t,2H),7.28(t,1H),7.18 -7.11(m,2H),7.07(t,1H),6.91(t,1H),6.81(d,1H),6.67(dd,1H),6.26(d, 1H),6.16(ddd,2H),5.62(d,1H),5.00(s,1H),4.31(ddd,1H),4.04(h,1H),3 .45(d,1H),3.39-3.33(m,3H),3.18-2.98(m,8H),2.94-2.84(m,2H),2.58(d t,1H),2.49(s,2H),2.16-2.3(m,3H),2.10(dq,1H),1.91-1.72(m,6H),1.63 -1.50(m,3H), 1.21(d,4H), 1.09 -1.05(m,2H). X-ray powder diffraction analysis revealed its 2θ as shown in Table 12, indicating that the product is D-mandelate A crystal form.

[0292] Example 10 Preparation of D-mandelate A crystal form

[0293] 100 mg of the compound shown in Formula I was dissolved in a mixed solution of 2.5 mL ethanol and 2.5 mL ethyl acetate. 21 mg of solid D-mandelic acid was then added and dissolved. The mixture was stirred overnight at room temperature until precipitation, centrifuged, and dried under vacuum to obtain the solid. X-ray powder diffraction analysis identified the product as mandelate A crystal form. The XRPD spectrum is shown below. Figure 6 As shown in Table 12, the positions of its characteristic peaks are as follows.

[0294] Example 11 Preparation of Oxalate I Crystal Form

[0295] 100 mg of the compound shown in Formula I was added to 2 mL of acetonitrile solvent. 24 mg of solid oxalic acid was added, but the mixture was insoluble. The mixture was stirred overnight at room temperature to obtain a white suspension. After centrifugation and vacuum drying, the solid was obtained. X-ray powder diffraction analysis identified the product as oxalate I crystal form. The XRPD spectrum is shown below. Figure 7 As shown in Table 13, the positions of its characteristic peaks are as follows. The DSC spectrum shows an endothermic peak at 136.67℃. The TGA spectrum shows a weight loss of 2.97% from 31 to 150℃. Ion chromatography results show an oxalate ion content of 18.7%.

[0296] Table 13

[0297]

[0298] Example 12 Preparation of Phosphate Amorphous Products

[0299] 100 mg of the compound shown in Formula I was added to 1 mL of acetone solvent, followed by 10 μL of 85% phosphoric acid solution. The mixture was stirred overnight at room temperature to obtain a white suspension. The suspension was centrifuged and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous phosphate compound, as shown in the XRPD pattern. Figure 8 As shown, the ion chromatography results indicate that the phosphate ion content is 12.51%.

[0300] Example 13 Preparation of Hydrochloride I Crystal Form

[0301] 50 mg of the compound shown in Formula I was added to 2 mL of acetonitrile solvent, followed by 11.5 μL of concentrated hydrochloric acid (36.5%). The mixture was stirred overnight at room temperature to obtain a white suspension. The suspension was centrifuged and dried under vacuum to obtain a solid. X-ray powder diffraction analysis identified the product as hydrochloride form I. The XRPD spectrum is shown below. Figure 9 As shown in Table 14, the positions of its characteristic peaks are as follows. The DSC spectrum shows an endothermic peak at 194.65℃. The TGA spectrum shows a weight loss of 9.15% from 31 to 200℃. Ion chromatography results show a chloride ion content of 7.2%.

[0302] Table 14

[0303]

[0304] Example 14 Preparation of Maleate Amorphous Compounds

[0305] 100 mg of the compound shown in Formula I was added to 2 mL of 2-methyl-tetrahydrofuran solvent, followed by 15 mg of solid maleic acid. The mixture was stirred at room temperature until clear, then 8 mL of n-heptane was added, and the mixture was stirred overnight at room temperature. After centrifugation, the solid was obtained by vacuum drying. X-ray powder diffraction analysis showed that the product was an amorphous maleate salt, as shown in the XRPD spectrum. Figure 10As shown in the figure, ion chromatography results indicate that the maleate ion content is 12.74%.

[0306] Example 15 Preparation of Fumarate Amorphous Compounds

[0307] 10 mg of the compound shown in formula (I) was added to 0.3 mL of tetrahydrofuran solvent, followed by 1.5 mg of fumaric acid. The mixture was stirred at room temperature until clear, then 1.0 mL of n-heptane was added, and the mixture was stirred overnight at room temperature. After centrifugation, the solid was obtained by vacuum drying. X-ray powder diffraction analysis showed that the product was an amorphous fumarate, as shown in the XRPD spectrum. Figure 11 As shown in the figure, ion chromatography results indicate that the fumarate ion content is 12.93%.

[0308] Example 16 Preparation of amorphous hydrobromide

[0309] 10 mg of the compound shown in Formula I was dissolved and clarified in 0.3 mL of acetone. Then, 1.5 μL of hydrobromic acid was added, and the mixture was stirred overnight at room temperature. After centrifugation, the solid was dried under vacuum. X-ray powder diffraction analysis showed that the product was an amorphous hydrobromide, as shown in the XRPD spectrum. Figure 12 As shown in the figure, ion chromatography results indicate a bromide ion content of 14.91%.

[0310] Example 17 Preparation of amorphous citrate

[0311] 10 mg of the compound shown in formula (I) was added to 0.3 mL of tetrahydrofuran solvent, followed by 1.5 mg of citric acid. The mixture was stirred at room temperature until clear, then 1.0 mL of n-heptane was added, and the mixture was stirred overnight at room temperature. After centrifugation, the solid was obtained by vacuum drying. X-ray powder diffraction analysis showed that the product was an amorphous citrate, as shown in the XRPD spectrum. Figure 13 As shown in the figure, ion chromatography results indicate that the citrate ion content is 19.61%.

[0312] Example 18 Preparation of sulfate amorphous products

[0313] 10 mg of the compound shown in Formula I was dissolved and clarified in 0.3 mL of acetonitrile solvent. Then, 0.70 μL of concentrated sulfuric acid was added, and the mixture was stirred overnight at room temperature. After centrifugation, the solid was obtained by vacuum drying. X-ray powder diffraction analysis showed that the product was an amorphous sulfate compound, as shown in the XRPD spectrum. Figure 14 As shown in the figure, ion chromatography results indicate that the sulfate ion content is 15.58%.

[0314] Example 19 Preparation of amorphous methanesulfonate

[0315] 10 mg of the compound shown in Formula I was added to 0.3 mL of water for suspension, followed by the addition of 0.85 μl of methanesulfonic acid. The mixture was reacted at room temperature for 2 h, then stirred overnight at 40 °C. After centrifugation, the solid was obtained by vacuum drying. X-ray powder diffraction analysis revealed that the product was an amorphous methanesulfonate, as shown in the XRPD spectrum. Figure 15 As shown.

[0316] Example 20 Preparation of hippurate amorphous product

[0317] 10 mg of the compound shown in Formula I was added to 2.48 mg of solid hippuric acid, dissolved in a mixture of 0.1 mL ethanol and 0.1 mL ethyl acetate, and precipitated by adding 700 μL of MTBE. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous hippuric acid salt, and the XRPD spectrum is shown below. Figure 16 As shown.

[0318] Example 21 Preparation of Benzoate Amorphous Compounds

[0319] 10 mg of the compound shown in Formula I was added to 1.69 mg of solid benzoic acid, then to 0.1 mL of acetonitrile or a mixture of 0.1 mL ethanol and 0.1 mL ethyl acetate. 700 μL of methyl tert-butyl ether was added to precipitate the product. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous benzoate, as shown in the XRPD pattern. Figure 17 As shown.

[0320] Example 22 Preparation of Benzoate Amorphous Products

[0321] 10 mg of the compound shown in Formula I was added to 1.69 mg of solid benzoic acid, then to 0.3 mL of methyl tert-butyl ether solvent. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction (XRPD) analysis showed no obvious characteristic peaks, indicating that the product was an amorphous benzoate.

[0322] Example 23 Preparation of amorphous malate

[0323] 10 mg of the compound shown in formula (I) was added to 1.85 mg of solid malic acid, then to a solution of 0.5 mL acetonitrile or a mixture of 0.1 mL ethanol and 0.1 mL ethyl acetate. 700 μL of methyl tert-butyl ether was added to precipitate the product. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous malate, as shown in the XRPD pattern. Figure 18 As shown.

[0324] Example 24 Preparation of amorphous malate

[0325] 10 mg of the compound shown in formula (I) was added to 1.85 mg of solid malic acid, then to 0.3 mL of tetrahydrofuran solvent. 700 μL of methyl tert-butyl ether was added to precipitate the product. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous malate, as shown in the XRPD spectrum. Figure 18 As shown.

[0326] Example 25 Preparation of amorphous acetate

[0327] 10 mg of the compound shown in formula (I) was added to 11.5 μl of a 1.2 M acetic acid-ethanol solution, dissolved in 0.3 mL of acetonitrile or ethyl acetate, and precipitated by adding 700 μL of methyl tert-butyl ether. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction analysis showed that the product was an amorphous acetate, and the XRPD spectrum is shown below. Figure 19 As shown.

[0328] Example 26 Preparation of amorphous acetate

[0329] 10 mg of the compound shown in Formula I was added to 11.5 μl of a 1.2 M acetic acid-ethanol solution, then added to 0.3 mL of a methyl tert-butyl ether solution. The mixture was stirred overnight at room temperature, centrifuged, and dried under vacuum to obtain a solid. X-ray powder diffraction (XRPD) analysis showed no obvious characteristic peaks, indicating that the product was an amorphous acetate.

[0330] Example 28: Stability Study of Amorphous Materials

[0331] Stability study of influencing factors: The free state samples were laid out in the open and the stability of the samples was investigated under high temperature (40℃, 60℃) and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days.

[0332] Table 15

[0333]

[0334] Experimental results show that the free amorphous state exhibits good chemical stability under high temperature and high humidity conditions.

[0335] Long-term / accelerated stability study: The free state sample was sealed in an aluminum foil bag and placed under conditions of 25℃ / 60%RH and 40℃ / 75%RH to investigate its stability. The results are shown below.

[0336] Table 16

[0337]

[0338] Experimental results show that the free amorphous state exhibits good physical and chemical stability after being placed under long-term accelerated conditions for 6 months.

[0339] Example 29 Salt Form Stability Study

[0340] Stability study of factors affecting salt type: Salt type samples were laid out in the open and the stability of the samples was investigated under high temperature (40℃, 60℃) and high humidity (RH 75%) conditions. The sampling period was 30 days.

[0341] Table 17

[0342]

[0343] Table 18

[0344]

[0345] Table 19

[0346]

[0347]

[0348] Long-term / accelerated stability of salt form: The samples were sealed in aluminum foil bags and subjected to stability tests under conditions of 25℃ / 60%RH and 40℃ / 75%RH, respectively. The results are shown below.

[0349] Table 20

[0350]

[0351] Experimental results show that succinate B crystal form and mandelate A crystal form exhibit good chemical and physical stability after 6 months of long-term accelerated storage. Maleate amorphous form also exhibits good chemical and physical stability after 6 months of long-term storage.

Claims

1. An amorphous form of the compound shown in Formula I, with a diffraction angle of 2... θ The X-ray powder diffraction pattern expressed in terms of angle is shown in Figure 1. 。 2. A pharmaceutically acceptable salt of the compound shown in Formula I, wherein the pharmaceutically acceptable salt is selected from succinate, mandelate, oxalate, phosphate, hydrochloride, maleate, fumarate, hydrobromide, citrate, sulfate, methanesulfonate, hippurate, benzoate, malate, and acetate. 。 3. The pharmaceutically acceptable salt of the compound of formula I according to claim 2, wherein the pharmaceutically acceptable salt is selected from succinate and mandelate.

4. A pharmaceutically acceptable salt of the compound of formula I according to claim 2, wherein the pharmaceutically acceptable salt is selected from succinate and D-mandelate.

5. A pharmaceutically acceptable salt of the compound of formula I according to claim 2, wherein the chemical ratio of the compound of formula I to an acid molecule or an acid radical is selected from 2:0.5 to 1:

3.

6. A pharmaceutically acceptable salt of the compound of formula I according to claim 2, wherein the chemical ratio of the compound of formula I to the acid molecule or acid anion is selected from 2:1 to 1:

3.

7. A pharmaceutically acceptable salt of the compound of formula I according to claim 2, wherein the chemical ratio of the compound of formula I to the acid molecule or acid anion is selected from 1:0.5, 1:1, 1:1.5 or 1:

2.

8. A succinate of the compound shown in Formula I, wherein, The chemical ratio of compound I to succinate or succinic acid is 2:1 to 1:

3. 。 9. The succinate of the compound of formula I according to claim 8, wherein, The chemical ratio of the compound of Formula I to succinate or succinic acid is selected from 1:1 or 1:

2.

10. A succinate B crystal form of the compound shown in Formula I, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 6.402, 9.400, 15.728, 17.957, and 19.

070. 。 11. The succinate B crystal form of the compound of formula I according to claim 10, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 6.402, 9.400, 15.728, 17.957, 19.070, and 21.

589.

12. The succinate B crystal form of the compound of formula I according to claim 10, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed in terms of angle, is shown in Figure 3.

13. A method for preparing the crystal form according to any one of claims 10-12, comprising: The compound shown in Formula I is mixed with a solvent and succinic acid, and crystallization is performed to obtain succinate B crystal form. The solvent is selected from ketone solvents.

14. The method for preparing the crystal form according to claim 13, wherein the ketone solvent is acetone.

15. A D-mandelate of the compound shown in Formula I, wherein, The chemical ratio of compound I to mandelic acid or mandelic acid is 1:

1. 。 16. A crystal form of D-mandelate of the compound shown in Formula I, wherein the crystal form is determined by a diffraction angle of 2°. θ The X-ray powder diffraction pattern, expressed as an angle, shows characteristic peaks at 10.554, 13.168, 15.728, 17.944, and 19.

784. 。 17. The D-mandelate A crystal form of the compound of formula I according to claim 16, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 6.159, 8.371, 10.554, 13.168, 15.728, 17.944, 19.784, and 20.

381.

18. The D-mandelate A crystal form of the compound of formula I according to claim 16, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed in terms of angle, shows characteristic peaks at 6.159, 8.371, 10.554, 13.168, 15.728, 17.944, 19.784, 20.381, 21.121, and 21.

434.

19. The D-mandelate A crystal form of the compound of formula I according to claim 16, wherein the diffraction angle is 2... θ The X-ray powder diffraction pattern, expressed in terms of angle, is shown in Figure 6.

20. A method for preparing the crystal form according to any one of claims 16-19, comprising: The compound shown in Formula I was mixed with a solvent and D-mandelic acid, and crystallized to obtain D-mandelic acid salt A crystal form; The solvent is selected from one or more of isopropanol, ethanol, and ethyl acetate.

21. The method for preparing the crystal form according to claim 20, wherein the solvent is selected from isopropanol / ethyl acetate, ethanol / ethyl acetate, and ethyl acetate.

22. A pharmaceutical composition comprising the amorphous material of claim 1, the salt of any one of claims 3-9, the crystalline form of any one of claims 10-12, the crystalline form of any one of claims 16-19 or the crystalline form prepared by any one of claims 13-14, 20-21, and optionally a pharmaceutically acceptable excipient.

23. A method for preparing a pharmaceutical composition according to claim 22, comprising the step of mixing the amorphous material of claim 1, the salt of any one of claims 3-9, the crystal form of any one of claims 10-12, the crystal form of any one of claims 16-19, or the crystal form obtained by the preparation method of any one of claims 13-14 and 20-21, with a pharmaceutically acceptable excipient.

24. Use of the amorphous material according to claim 1, the salt according to any one of claims 3-9, the crystal form according to any one of claims 10-12, the crystal form according to any one of claims 16-19, the crystal form obtained by the preparation method according to any one of claims 13-14, 20-21, or the pharmaceutical composition according to claim 22 in the preparation of a medicament for regulating the ubiquitination and degradation of androgen receptor (AR) proteins.

25. Use of the amorphous material according to claim 1, the salt according to any one of claims 3-9, the crystal form according to any one of claims 10-12, the crystal form according to any one of claims 16-19, the crystal form obtained by the preparation method according to any one of claims 13-14, 20-21, or the pharmaceutical composition according to claim 22 in the preparation of a medicament for treating and / or preventing androgen receptor-mediated or dependent diseases or conditions.

26. The use according to claim 25, wherein the androgen receptor-mediated or dependent disease or condition is selected from tumors, male sexual dysfunction, and Kennedy's disease.

27. The use according to claim 25, wherein the androgen receptor-mediated or dependent disease or condition is selected from prostate cancer, benign prostatic hyperplasia, hirsutism, alopecia, anorexia nervosa, breast cancer, acne, male sexual dysfunction, Kennedy's disease, and AIDS.

28. The use according to claim 25, wherein the androgen receptor-mediated or dependent disease or condition is selected from prostate cancer.

29. The use according to claim 25, wherein the androgen receptor-mediated or dependent disease or condition is selected from hormone-sensitive prostate cancer or hormone-refractory prostate cancer.

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