A salt of an isoquinolinone compound and a crystal form thereof

Abstract

The present disclosure relates to a salt of isoquinolinone compound and a crystal form thereof. Specifically, the present disclosure relates to a pharmaceutically acceptable salt of a sulfur-containing isoindoline derivative, a crystal form thereof, and a preparation method thereof. Provided are p-toluenesulfonic acid salt, methanesulfonic acid salt, phosphoric acid salt, hydrobromic acid salt, and hydrochloric acid salt of a compound as shown in Formula I, and a crystalline form.

Description

Technical Field

[0001] This disclosure relates to a pharmaceutically acceptable salt, crystal form, and preparation method of a sulfur-containing isoindoline derivative, specifically providing p-toluenesulfonate, methanesulfonate, phosphate, hydrobromide, and hydrochloride salts of compounds as shown in formula (I), as well as their crystalline forms. Background Technology

[0002] Multiple myeloma (MM) is a malignant tumor with main symptoms including hypercalcemia, kidney damage, anemia, and bone disease. MM is the second most common hematologic malignancy after non-Hodgkin's lymphoma, and current treatments primarily involve drug therapy and autologous stem cell transplantation.

[0003] Currently, there are four main classes of drugs widely used in clinical practice: lenalidomide immunomodulators, proteasome inhibitors, hormones, and monoclonal antibodies. Drugs in clinical research stages include bispecific antibodies, ADCs, and CAR-T therapy. These drugs have different mechanisms of action, and combination therapy often achieves better efficacy. Clinically, dual, triple, or even quadruple therapy is commonly used, typically combining immunomodulators, proteasome inhibitors, and hormones, sometimes with the addition of antibodies. Lenalidomide is the most commonly used immunomodulator, used in first-line treatment, maintenance therapy after stem cell transplantation, and second- and third-line treatment after relapse. Its sales reached $9.7 billion in 2018 / 2019. The entire MM market is also considerable and growing rapidly, due to continuous improvements in the diagnosis and treatment of MM, leading to longer patient survival and extended treatment durations. The MM market is projected to reach $33 billion by 2022, with immunomodulators, represented by lenalidomide, still accounting for the largest share.

[0004] The mechanism of action of immunomodulators (IMiDs) in treating multiple myeloma (MM) is primarily based on the fact that IMiD drugs bind to the Cerebrolysin B (CRBN) protein, activating the E3 ligase activity of CRBN, which then selectively binds to the transcription factors Ikaros (IKZF1) and Aiolos (IKZF3). This leads to the rapid ubiquitination and degradation of Ikaros and Aiolos. Downregulation of Ikaros / Aiolos results in downregulation of c-Myc, followed by downregulation of IRF4, ultimately inhibiting myeloma cell growth and inducing apoptosis. Furthermore, IKZF3 can inhibit the transcription of IL-2 and TNF cytokines in T / NK cells. IKZF3 degradation relieves this inhibition, promoting the release of these cytokines and thus exerting an immunomodulatory effect. Clinical trials have also shown a correlation between the clinical benefits of IMiD drugs and the level of CRBN expression. Knocking down CRBN in lenalidomide-sensitive cell lines (OPM2 and KMS18) revealed the loss of lenalidomide's inhibitory activity on cell growth, leading to drug resistance. The level of CRBN knockdown was correlated with the degree of drug resistance. In cell proliferation experiments, reducing the expression level of CRBN in cells (U266-CRBN60 and U266-CRBN75) reduced the inhibitory activity of both lenalidomide and pomalidomide on cell growth.

[0005] Currently approved IMiD drugs include thalidomide, lenalidomide, and pomalidomide, all from Celgene (now merged with BMS). The binding affinity of the three compounds to CRBN increases sequentially, hence the clinical dosage decreases accordingly. The primary indication for all three compounds is multidisciplinary disease (MM). Thalidomide and lenalidomide also have other indications, especially lenalidomide, which can be used to treat myelodysplastic syndromes (MDS). Regarding side effects, lenalidomide and pomalidomide exhibit similar effects, with significant myelosuppression, a target-related toxicity. Thalidomide has some other side effects, such as sedation, constipation, and neurological side effects.

[0006] All IMiDs' adipicimide moieties bind to a hydrophobic bag defined by three tryptophan residues in CRBN (called the "thalidomide binding bag"). Conversely, the phthalimide / isoindolone ring is exposed to the solvent and alters the molecular surface of CRBN, thereby modulating substrate recognition. Different IMiDs lead to significant modifications on the CRBN molecular surface and different substrate recognition preferences. Therefore, modifications to IMiDs may lead to the degradation of other transcription factors, causing unwanted toxic side effects. This mode of action of IMiDs is also known as molecular glue, vividly describing the binding effect of this small molecule on two protein substrates.

[0007] Because the median survival for multiple myeloma is currently over five years, this prolonged survival has led to a high proportion of patients developing resistance to currently marketed drugs such as lenalidomide and pomalidomide, severely reducing the effectiveness of these treatments. Therefore, we envision developing more active drug molecules to overcome drug resistance while minimizing the toxic side effects of these compounds.

[0008] Published patent applications for Cereblon modifiers include WO2008115516A2, WO2011100380A1, WO2019226770A1, WO2019014100A1 and WO2020064002A1, etc.

[0009] The applicant's patent application PCT / CN2021 / 107297 provides a compound as shown in Formula I, with the chemical name (S)-4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile. This compound exhibits good Cereblon modulating activity.

[0010] Summary of the Invention

[0011] This disclosure provides pharmaceutically acceptable salts of compounds represented by formula (I), wherein the pharmaceutically acceptable salts are selected from p-toluenesulfonate, methanesulfonate, phosphate, hydrobromide, and hydrochloride, and the chemical name of the compound represented by formula (I) is (S)-4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile,

[0012]

[0013] In some embodiments, the chemical ratio of the compound represented by formula (I) to the acid molecule is 5:1 to 1:5, preferably 2:1, 1:1, or 1:2, and most preferably 2:1, 1:1, or 1:2. In some embodiments, the chemical ratio of the compound represented by formula (I) to p-toluenesulfonic acid molecules is 1:1 or 2:1. In some embodiments, the chemical ratio of the compound represented by formula (I) to methanesulfonic acid molecules is 1:1. In some embodiments, the chemical ratio of the compound represented by formula (I) to phosphoric acid is 1:1. In some embodiments, the chemical ratio of the compound represented by formula (I) to hydrochloric acid is 1:1. In some embodiments, the chemical ratio of the compound represented by formula (I) to hydrobromic acid is 1:1.

[0014] This disclosure provides a method for preparing a pharmaceutically acceptable salt of the compound of formula (I), comprising the step of reacting the compound of formula (I) with an acid to form a salt. In some embodiments, the solvent used in the salt-forming reaction is selected from one or more of acetonitrile, tetrahydrofuran, water, and acetone.

[0015] In some embodiments, the method for preparing the aforementioned pharmaceutically acceptable salt also includes steps such as evaporating solvents or stirring to crystallize, filtering, and drying.

[0016] This disclosure provides a pharmaceutical composition prepared from the aforementioned pharmaceutically acceptable salt.

[0017] This disclosure provides a pharmaceutical composition comprising the aforementioned pharmaceutically acceptable salt or a pharmaceutically acceptable salt prepared by the aforementioned method, and optionally a pharmaceutically acceptable carrier, diluent, or excipient.

[0018] This disclosure provides a method for preparing a pharmaceutical composition, comprising the step of mixing the aforementioned pharmaceutically acceptable salt, or a pharmaceutically acceptable salt of a compound of formula (I) prepared by the aforementioned method, with a pharmaceutically acceptable carrier, diluent, or excipient.

[0019] This disclosure provides the use of a pharmaceutically acceptable salt of the compound represented by formula (I) above, or a pharmaceutically acceptable salt prepared by the aforementioned method, or the aforementioned composition, or the composition prepared by the aforementioned method, in the preparation of a medicament for treating and / or preventing diseases related to CRBN protein.

[0020] This disclosure provides the use of a pharmaceutically acceptable salt of the compound shown in formula (I) above, or a pharmaceutically acceptable salt prepared by the aforementioned method, or the aforementioned composition, or a composition prepared by the aforementioned method, in the preparation of a medicament for treating and / or preventing cancer, angiogenesis-related conditions, pain, macular degeneration or related syndromes, skin diseases, lung diseases, asbestos-related diseases, parasitic diseases, immunodeficiency diseases, CNS diseases, CNS damage, atherosclerosis or related conditions, sleep disorders or related conditions, infectious diseases, hemoglobinopathies or related conditions, or TNFα-related conditions; preferably, in the preparation of a medicament for treating and / or preventing cancer or CNS damage.

[0021] In some embodiments, the cancer is selected from leukemia, myeloma, lymphoma, melanoma, skin cancer, liver cancer, kidney cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, esophageal cancer, colorectal cancer, gallbladder cancer, bile duct cancer, choriocarcinoma, pancreatic cancer, polycythemia vera, pediatric tumors, cervical cancer, ovarian cancer, breast cancer, bladder cancer, urothelial carcinoma, ureteral tumors, prostate cancer, seminoma, testicular tumors, head and neck tumors, head and neck squamous cell carcinoma, endometrial cancer, thyroid cancer, sarcoma, osteoma, neuroblastoma, neuroendocrine carcinoma, brain tumor, CNS cancer, astrocytoma, and glioma; preferably, the liver cancer is hepatocellular carcinoma; the colorectal cancer is colon cancer or rectal cancer; the sarcoma is osteosarcoma or soft tissue sarcoma; and the glioma is glioblastoma.

[0022] In some embodiments, the myeloma is multiple myeloma (MM) and myelodysplastic syndrome (MDS); preferably, the multiple myeloma is relapsed, refractory, or resistant.

[0023] In some embodiments, the multiple myeloma is lenalidomide or pomalidomide refractory or resistant.

[0024] This disclosure provides the α-crystal form of the compound (I) p-toluenesulfonate, and its X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, showing characteristic peaks at 5.787, 6.961, 9.961, 15.756, 21.717, 23.539, and 26.272. In some embodiments, the X-ray powder diffraction pattern of the α-crystal form of the compound (I) p-toluenesulfonate, expressed in terms of diffraction angle 2θ, is as follows: Figure 1 As shown.

[0025] This disclosure further provides a method for preparing the a-crystal form of the compound shown in formula (I), comprising: method 1, a) mixing the compound shown in formula (I), acetonitrile and a p-toluenesulfonic acid solution, heating, b) cooling to crystallize.

[0026] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 200. In some embodiments, the method for preparing the α-crystal form of p-toluenesulfonate according to this disclosure further includes steps such as filtration, washing, or drying.

[0027] This disclosure provides the b-crystal form of the p-toluenesulfonate compound of formula (I), and its X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, showing characteristic peaks at 5.823, 11.447, 12.455, 13.772, 17.807, 22.501, and 23.072. In some embodiments, the X-ray powder diffraction pattern of the b-crystal form of the p-toluenesulfonate compound of formula (I), expressed in terms of diffraction angle 2θ, is as follows: Figure 2 As shown.

[0028] This disclosure further provides a method for preparing the b-crystal form of the compound shown in formula (I), comprising: a) mixing the compound shown in formula (I) with a solution of p-toluenesulfonic acid and tetrahydrofuran, and heating; b) cooling to crystallize.

[0029] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 200. In some embodiments, the method for preparing p-toluenesulfonate b crystal form according to this disclosure further includes steps such as filtration, washing, or drying.

[0030] This disclosure provides the α-crystal form of the methanesulfonate of formula (I), with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 8.616, 11.915, 17.525, 19.197, 22.233, 24.561, and 25.679. In some embodiments, the α-crystal form of the methanesulfonate of formula (I) has characteristic peaks at 8.616, 11.915, 17.525, 19.197, 19.969, 22.233, 24.124, 24.561, 25.679, and 27.524. In some embodiments, the α-crystal form of the methanesulfonate compound shown in formula (I) exhibits characteristic peaks at 8.616, 11.915, 17.525, 19.197, 19.969, 22.233, 24.124, 24.561, 25.679, 27.524, 28.725, 31.000, and 34.434. In some embodiments, the X-ray powder diffraction pattern of the α-crystal form of the methanesulfonate compound shown in formula (I), expressed in terms of diffraction angle 2θ, is shown below. Figure 3 As shown.

[0031] This disclosure further provides a method for obtaining the a-crystal form of the methanesulfonate of the compound shown in formula (I), comprising: a) mixing the compound shown in formula I with acetonitrile and a methanesulfonic acid solution, heating, and b) cooling to crystallize.

[0032] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 200. In some embodiments, the method for preparing the α-crystal form of methanesulfonate according to this disclosure further includes steps such as filtration, washing, or drying.

[0033] This disclosure provides the b-crystal form of the methanesulfonate of formula (I), with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, and 24.655. In some embodiments, the b-crystal form of the methanesulfonate of formula (I) has characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, 24.655, 26.357, 29.109, and 32.893. In some embodiments, the b-crystal form of the methanesulfonate compound shown in formula (I) exhibits characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, 24.655, 26.357, 29.109, 32.893, and 35.688. In some embodiments, the X-ray powder diffraction pattern of the b-crystal form of the methanesulfonate compound shown in formula (I), expressed in terms of diffraction angle 2θ, is shown below. Figure 4 As shown.

[0034] This disclosure further provides a method for obtaining the b-crystal form of the methanesulfonate of the compound shown in formula (I), comprising the steps of: a) mixing the compound shown in formula (I) with a solution of tetrahydrofuran and methanesulfonic acid, and heating; b) cooling to crystallize.

[0035] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 200. In some embodiments, the method for preparing the b-crystal form of methanesulfonate according to this disclosure further includes steps such as filtration, washing, or drying.

[0036] This disclosure provides the c-crystal form of the methanesulfonate compound of formula (I), and its X-ray powder diffraction pattern, expressed in terms of diffraction angle 2θ, showing characteristic peaks at 11.191, 16.225, 17.493, 19.622, 22.683, 24.308, and 26.271. In some embodiments, the c-crystal form of the methanesulfonate compound of formula (I) shows characteristic peaks at 11.191, 15.027, 16.225, 17.493, 19.622, 22.683, 24.308, 25.113, and 26.271. In some embodiments, the X-ray powder diffraction pattern of the c-crystal form of the methanesulfonate compound of formula (I), expressed in terms of diffraction angle 2θ, is as follows: Figure 5 As shown.

[0037] This disclosure further provides a method for obtaining the c-crystal form of the methanesulfonate of the compound shown in formula (I), comprising the steps of: placing the b-crystal form of the methanesulfonate of the compound shown in formula (I) under 75% RH conditions for 30 days or after DVS.

[0038] This disclosure provides the α-crystal form of the phosphate compound of formula (I), with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 8.525, 12.180, 14.251, 17.952, 20.210, 22.157, and 27.662. In some embodiments, the α-crystal form of the phosphate compound of formula (I) has characteristic peaks at 8.525, 12.180, 14.251, 17.952, 19.282, 20.210, 22.157, 24.560, 25.821, and 27.662. In some embodiments, the α-crystal form of the phosphate compound shown in formula (I) exhibits characteristic peaks at 8.525, 12.180, 14.251, 16.158, 17.952, 19.282, 20.210, 22.157, 24.560, 25.404, 25.821, 27.662, and 28.781. In some embodiments, the X-ray powder diffraction pattern of the α-crystal form of the phosphate compound shown in formula (I), expressed in terms of diffraction angle 2θ, is as follows: Figure 6 As shown.

[0039] This disclosure further provides a method for obtaining the a-crystal form of the phosphate of the compound shown in formula (I), comprising the steps of: a) mixing the compound shown in formula (I) with acetonitrile and phosphoric acid solution, and heating; b) cooling to crystallize.

[0040] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 200. In some embodiments, the method for preparing the α-crystal form of phosphates described in this disclosure further includes steps such as filtration, washing, or drying.

[0041] This disclosure provides the α-crystal form of the hydrobromide of formula (I), with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, showing characteristic peaks at 9.405, 10.749, 17.979, 19.610, 20.887, 22.693, and 26.133. In some embodiments, the α-crystal form of the hydrobromide of formula (I) has characteristic peaks at 9.405, 10.749, 12.992, 15.520, 17.979, 19.610, 20.887, 22.693, 25.458, and 26.133. In some embodiments, the α-crystal form of the hydrobromide of formula (I) exhibits characteristic peaks at 9.405, 10.749, 12.992, 15.520, 17.979, 19.610, 20.887, 22.693, 25.458, 26.133, 27.099, 29.346, and 31.679. In some embodiments, the X-ray powder diffraction pattern of the α-crystal form of the hydrobromide of formula (I), expressed in terms of diffraction angle 2θ, is shown below. Figure 7 As shown.

[0042] This disclosure further provides a method for obtaining the a-crystal form of the hydrobromide of the compound shown in formula (I), comprising the steps of: a) mixing the compound shown in formula (I) with a hydrobromic acid solution, a solvent selected from acetonitrile, tetrahydrofuran and water / acetone, and heating to dissolve; b) cooling to crystallize.

[0043] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 200. In some embodiments, the method for preparing the α-crystal form of hydrobromide described in this disclosure further includes steps such as filtration, washing, or drying.

[0044] This disclosure provides the α-crystal form of the hydrochloride salt of the compound shown in formula (I), with an X-ray powder diffraction pattern expressed as a diffraction angle 2θ, exhibiting characteristic peaks at 7.019, 9.760, 10.683, 17.988, 21.349, 23.050, and 26.039. In some embodiments, the α-crystal form of the hydrochloride salt of the compound shown in formula (I) exhibits characteristic peaks at 7.019, 9.760, 10.683, 13.645, 16.287, 17.988, 19.646, 21.349, 23.050, and 26.039. In some embodiments, the α-crystal form of the hydrochloride salt of formula (I) exhibits characteristic peaks at 7.019, 9.760, 10.683, 12.844, 13.645, 16.287, 17.988, 19.646, 21.349, 23.050, and 26.039. In some embodiments, the X-ray powder diffraction pattern of the α-crystal form of the hydrochloride salt of formula (I), expressed in terms of diffraction angle 2θ, is as follows: Figure 8 As shown.

[0045] This disclosure further provides a method for obtaining the a-crystal form of the hydrochloride salt of the compound shown in formula (I), comprising the steps of: a) mixing the compound shown in formula (I) with a hydrochloric acid solution, a solvent selected from acetonitrile, tetrahydrofuran and water / acetone, and heating; b) cooling to crystallize.

[0046] In some embodiments, the volume (μl) of the solvent used in this disclosure may be 1-200 times the mass (mg) of the compound of formula (I), and in non-limiting embodiments may be 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 200. In some embodiments, the method for preparing the α-crystal form of hydrochloride described in this disclosure further includes steps such as filtration, washing, or drying.

[0047] This disclosure also provides pharmaceutical compositions prepared from the crystal form of the pharmaceutically acceptable salt of the compound shown in formula (I).

[0048] This disclosure also provides a pharmaceutical composition comprising the crystal form of the aforementioned pharmaceutically acceptable salt and optionally a pharmaceutically acceptable carrier, diluent, or excipient.

[0049] This disclosure also provides a method for preparing a pharmaceutical composition, comprising the step of mixing the crystal form of the aforementioned pharmaceutically acceptable salt with a pharmaceutically acceptable carrier, diluent, or excipient.

[0050] This disclosure also provides the use of the crystal form of the aforementioned pharmaceutically acceptable salt, or the aforementioned composition, or the composition prepared by the aforementioned method, in the preparation of a medicament for treating and / or preventing diseases related to CRBN protein.

[0051] This disclosure also provides the use of the aforementioned pharmaceutically acceptable salt crystal form, or the aforementioned composition, or the composition prepared by the aforementioned method, in the preparation of medicaments for the treatment and / or prevention of cancer, angiogenesis-related conditions, pain, macular degeneration or related syndromes, skin diseases, lung diseases, asbestos-related diseases, parasitic diseases, immunodeficiency diseases, CNS diseases, CNS damage, atherosclerosis or related conditions, sleep disorders or related conditions, infectious diseases, hemoglobinopathies or related conditions, or TNFα-related conditions; preferably, in the preparation of medicaments for the treatment and / or prevention of cancer or CNS damage.

[0052] In some embodiments, the cancer is selected from leukemia, myeloma, lymphoma, melanoma, skin cancer, liver cancer, kidney cancer, lung cancer, nasopharyngeal carcinoma, gastric cancer, esophageal cancer, colorectal cancer, gallbladder cancer, bile duct cancer, choriocarcinoma, pancreatic cancer, polycythemia vera, pediatric tumors, cervical cancer, ovarian cancer, breast cancer, bladder cancer, urothelial carcinoma, ureteral tumors, prostate cancer, seminoma, testicular tumors, head and neck tumors, head and neck squamous cell carcinoma, endometrial cancer, thyroid cancer, sarcoma, osteoma, neuroblastoma, neuroendocrine carcinoma, brain tumor, CNS cancer, astrocytoma, and glioma; preferably, the liver cancer is hepatocellular carcinoma; the colorectal cancer is colon cancer or rectal cancer; the sarcoma is osteosarcoma or soft tissue sarcoma; and the glioma is glioblastoma.

[0053] In some embodiments, the myeloma is multiple myeloma (MM) and myelodysplastic syndrome (MDS); preferably, the multiple myeloma is relapsed, refractory, or resistant.

[0054] In some embodiments, the multiple myeloma is lenalidomide or pomalidomide refractory or resistant.

[0055] The determination of the stoichiometry of the compound of formula (I) described in this disclosure with the base or acid molecule is subject to a certain error. Generally, ±10% is within a reasonable error range. The error may vary to a certain extent depending on the context in which it is used, but this variation shall not exceed ±10%, ±9%, ±8%, ±7%, ±6%, ±5%, ±4%, ±3%, ±2%, ±1%, preferably ±5%.

[0056] The "2θ or 2θ angle" mentioned in this disclosure refers to the diffraction angle, where θ is the Bragg angle, and the unit is ° or degree; the error range of 2θ for each characteristic peak is ±0.20 (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.

[0057] 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,

[0058] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0059] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0060] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0061] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0062] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0063] 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.

[0064] The drying temperature described in this disclosure is generally 25℃~150℃, preferably 40℃~80℃, and can be dried under normal pressure or reduced pressure.

[0065] "Pharmaceutical composition" means a mixture containing one or more compounds of formula (I) described herein, or pharmaceutically acceptable salts thereof, along with other chemical components, such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and its biological activity.

[0066] The crystal forms described in this disclosure include, but are not limited to, solvates of pharmaceutically acceptable salts of compounds of formula (I), wherein the solvents include, but are not limited to, acetonitrile, tetrahydrofuran, and water / acetone.

[0067] The “solvents” described in this disclosure include, but are not limited to, complexes formed by combining pharmaceutically acceptable salts of compounds of formula (I) with solvents. Attached Figure Description

[0068] Figure 1 XRPD pattern of p-toluenesulfonate a crystal form of the compound shown in formula (I).

[0069] Figure 2 XRPD pattern of p-toluenesulfonate b crystal form of compound (I).

[0070] Figure 3 XRPD pattern of the crystal form of methanesulfonate a shown in formula (I).

[0071] Figure 4 XRPD pattern of the methanesulfonate b crystal form of the compound shown in formula (I).

[0072] Figure 5 XRPD pattern of the c-crystal form of the compound shown in formula (I).

[0073] Figure 6 XRPD pattern of phosphate a crystal form of the compound shown in formula (I).

[0074] Figure 7 XRPD pattern of hydrobromide a crystal form of the compound shown in formula (I).

[0075] Figure 8 XRPD pattern of the a-crystal form of the compound shown in formula (I).

[0076] Figure 9 Example 1: Efficacy data of compound CC-92480 and control CC-92480 against NCI-H929 xenografts in CB-17SCID mice.

[0077] Figure 10 Example 1: Effects of compound CC-92480 and control CC-92480 on body weight of CB-17SCID mice. Detailed Implementation Plan

[0078] The present disclosure will be explained in more detail below with reference to embodiments or experimental examples. The embodiments or experimental examples in the present disclosure are only used to illustrate the technical solutions in the present disclosure and are not intended to limit the substance and scope of the present disclosure.

[0079] Experimental methods in the embodiments of this disclosure that do not specify specific conditions are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose specific source is not specified are commercially available conventional reagents.

[0080] 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 measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard.

[0081] MS measurements were performed using an Agilent 1200 / 1290DAD-6110 / 6120 Quadrupole MS LC-MS system (manufacturer: Agilent, MS model: 6110 / 6120 Quadrupole MS), a Waters ACQuity UPLC-QD / SQD system (manufacturer: Waters, MS model: Waters ACQuity Qda Detector / Waters SQ Detector), or a THERMO Ultimate3000-Q Exactive system (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0082] High-performance liquid chromatography (HPLC) analysis was performed using an Agilent HPLC 1200DAD, an Agilent HPLC 1200VWD, and a Waters HPLC e2695-2489 high-performance liquid chromatograph.

[0083] Chiral HPLC analysis was performed using an Agilent 1260DAD high-performance liquid chromatograph.

[0084] High performance liquid chromatography (HPLC) was performed using Waters 2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP, and Gilson-281 preparative chromatographs.

[0085] Chiral preparation was performed using a Shimadzu LC-20AP preparative chromatograph.

[0086] The CombiFlash rapid preparation system uses a CombiFlash Rf200 (TELEDYNE ISCO).

[0087] 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.

[0088] Silica gel column chromatography generally uses Yantai Huanghai silica gel with a mesh size of 200-300 as the carrier.

[0089] Mean inhibition rate of kinases and IC 50 The values ​​were determined using a NovoStar microplate reader (BMG GmbH, Germany).

[0090] 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.

[0091] Unless otherwise specified in the examples, the reactions can be carried out under an argon or nitrogen atmosphere.

[0092] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0093] A hydrogen atmosphere refers to a reaction flask connected to a hydrogen balloon with a volume of approximately 1L.

[0094] The pressurized hydrogenation reaction was performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0095] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0096] The microwave reaction was performed using a CEM Discover-S 908860 microwave reactor.

[0097] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0098] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0099] 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: n-hexane / ethyl acetate system, B: dichloromethane / methanol 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.

[0100] The testing conditions of the instruments used in the experiments in this disclosure are as follows:

[0101] 1. Differential Scanning Calorimeter (DSC)

[0102] Instrument Model: Mettler Toledo DSC 3+STARe System

[0103] Purging gas: nitrogen; Nitrogen purging rate: 50 mL / min

[0104] Heating rate: 10.0℃ / min

[0105] Temperature range: 25-350℃ (or 25℃-300℃)

[0106] 2. X-ray Powder Diffraction (XRPD)

[0107] Instrument Model: BRUKER D8 Discover X-ray Powder Diffractometer

[0108] Rays: Monochromatic Cu-Kα rays

[0109] Scanning mode: θ / 2θ, scanning range (2θ range): 3~50°

[0110] Voltage: 40kV, Current: 40mA

[0111] 3. Thermogravimetric Analysis (TGA)

[0112] Instrument model: Mettler Toledo TGA2

[0113] Purging gas: nitrogen; Nitrogen purging rate: 50 mL / min

[0114] Heating rate: 10.0℃ / min

[0115] Temperature range: 25-400℃ (or 25℃-350℃)

[0116] 4. DVS is a dynamic moisture adsorption method.

[0117] The test was performed using SMS DVS Advantage at 25°C with humidity ranging from 0% to 95% in 10% increments. The judgment criterion was that the mass change dM / dT for each gradient was less than 0.002%, with a TMAX of 360 min and two cycles.

[0118] 5. Anion chromatography

[0119] Instrument Model: DIONEX INTEGRION HPIC Ion Chromatograph (USA)

[0120] Detection method: conductivity; Separation column: Dionex IonPac TM -AS11-HC

[0121] Rinse solution: EGC-500-KOH

[0122] Flow rate: 1.5 ml / min

[0123] Example 1: Preparation of the compound shown in formula (I) (S)-4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile (Compound 1)

[0124]

[0125] first step

[0126] 4-((5-formylpyridin-2-yl)thio)piperidine-1-carboxylic acid tert-butyl ester 1b

[0127] Compound 1-1a (700 mg, 3.22 mmol), 6-fluoropyridine-3-carboxaldehyde 1a (443 mg, 3.54 mmol), and potassium carbonate (1.11 g, 8.05 mmol) were added to N,N-dimethylformamide (10 mL). The reaction mixture was heated to 80 °C and reacted for 1 hour. The reaction mixture was diluted with water (30 mL) and then extracted with ethyl acetate (40 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography using eluent system B to give title compound 1b (1.0 g, yield: 96%).

[0128] MS m / z (ESI): 267.1 [M-55].

[0129] Step 2

[0130] 6-(piperidin-4-ylthio)nicotinaldehyde trifluoroacetate 1c

[0131] Compound 1b (950 mg, 2.95 mmol) was dissolved in dichloromethane (10 mL), and trifluoroacetic acid (2 mL) was slowly added under ice bath conditions. The reaction mixture was reacted for 1 hour. The reaction solution was concentrated and dried to give the title compound 1c. The crude product was used directly in the next reaction without purification.

[0132] MS m / z (ESI): 223.1 [M+1].

[0133] Step 3

[0134] 3-Fluoro-4-(4-(((5-formylpyridin-2-yl)thio)piperidin-1-yl)benzonitrile 1d

[0135] Compound 1c (760 mg, 2.94 mmol), 3,4-difluorobenzonitrile (817 mg, 5.87 mmol), and potassium carbonate (1.22 g, 8.81 mmol) were added to N,N-dimethylformamide (15 mL), and the reaction mixture was heated to 80 °C and reacted overnight. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography using eluent system B to give title compound 1d (850 mg, yield: 84%).

[0136] MS m / z (ESI): 342.1 [M+1].

[0137] Step 4

[0138] 3-Fluoro-4-(4-((5-(hydroxymethyl)pyridin-2-yl)thio)piperidin-1-yl)benzonitrile 1e

[0139] Compound 1d (600 mg, 1.76 mmol) was added to methanol (10 mL) under ice bath conditions, followed by slow addition of sodium borohydride (133 mg, 3.51 mmol), and the reaction was allowed to proceed for 1 hour. The reaction mixture was quenched with water (10 mL) and then extracted with ethyl acetate (30 mL × 3). The combined organic phases were washed with saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography using eluent system B to give the title compound 1e (580 mg, yield: 96%).

[0140] MS m / z (ESI): 344.1 [M+1].

[0141] Step 5

[0142] 4-(4-((5-(bromomethyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile 1f

[0143] Compound 1e (200 mg, 0.582 mmol) was added to dichloromethane (6 mL), followed by the addition of triphenylphosphine (199 mg, 0.757 mmol) and carbon tetrabromide (251 mg, 0.757 mmol). The reaction mixture was reacted for 2 hours. The reaction solution was concentrated, and the residue was purified by column chromatography using eluent system B to give the title compound 1f (190 mg, yield: 80%).

[0144] MS m / z(ESI): 406.0[M+1]; 408.0[M+3].

[0145] Step 6

[0146] (S)-5-amino-4-(4-((6-((1-(4-cyano-2-fluorophenyl)piperidin-4-yl)thio)pyridin-3-yl)methoxy)-1-oxoisoindolin-2-yl)-5-oxovalerate tert-butyl ester 1g

[0147] (S)-5-amino-4-(4-hydroxy-1-oxoisoindoline-2-yl)-5-oxovalerate tert-butyl ester 1-3a (82 mg, 0.246 mmol, prepared by a known method, "Journal of Medicinal Chemistry, 2020, 63(13), 6648-6676") and anhydrous potassium carbonate (65 mg, 0.468 mmol) were added to N,N-dimethylformamide (3 mL), and compound 1f (95 mg, 0.244 mmol) was added. The reaction mixture was reacted for 2 hours. The reaction mixture was poured into ice water (10 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL × 2), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography using eluent system B to give 1 g (145 mg, yield: 94%) of the title compound.

[0148] MS m / z (ESI): 660.2 [M+1].

[0149] Step 7 (S)-4-(4-((5-(((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-4-yl)oxy)methyl)pyridin-2-yl)thio)piperidin-1-yl)-3-fluorobenzonitrile 1

[0150] 1 g (60 mg, 0.091 mmol) of compound was added to acetonitrile (5 mL), and benzenesulfonic acid (16 mg, 0.091 mmol) was added at room temperature. The reaction mixture was reacted at 80 °C for 8 hours. The solvent was removed from the reaction solution under reduced pressure, and the residue was prepared by high performance liquid chromatography (Gilson GX-281, elution system: 10 mmol / L ammonium bicarbonate aqueous solution and acetonitrile, acetonitrile gradient: 60%-80%, flow rate: 30 mL / min) to give title compound 1 (42 mg, yield: 78%).

[0151] X-ray powder diffraction analysis showed that title compound 1 is amorphous. MS m / z (ESI): 586.4 [M+1].

[0152] 1 H NMR (500MHz, DMSO-d6): δ10.98(s,1H),8.60(s,1H),7.78(dd,1H),7.69(dd,1H),7.59-7. 48(m,2H),7.38-7.33(m,3H),7.16(t,1H),5.24(s,2H),5.12(dd,1H),4.42(d,1H),4.26( d,1H),4.07-3.99(m,1H),3.56-3.48(m,2H),3.13-3.03(m,2H),2.96-2.87(m,1H),2.63- 2.55(m,1H),2.46-2.38(m,1H),2.21-2.12(m,2H),2.02-1.94(m,1H),1.82-1.70(m,2H).

[0153] Biological evaluation

[0154] Test Example 1: Biological Evaluation of NCI-H929 Proliferation Experiment

[0155] The following method was used to determine the inhibitory activity of the disclosed compound on the proliferation of NCI-H929 cells. The experimental method is briefly described below.

[0156] NCI-H929 cells (ATCC, CRL-9068) were cultured in complete medium containing 10% fetal bovine serum (Corning, 35-076-CV) and 0.05 mM 2-mercaptoethanol (Sigma, M3148) in RPMI 1640 medium (Hyclone, SH30809.01). On day 1, NCI-H929 cells were seeded at a density of 6000 cells / well in 96-well plates using complete medium, with 100 μL of cell suspension per well. Simultaneously, 10 μL of serially diluted test compounds prepared in complete medium were added to each well. The compounds were first dissolved in DMSO at an initial concentration of 10 mM, and then serially diluted 5-fold to a total of 9 concentrations. The blank control was 100% DMSO. Then, 5 μL of the DMSO-dissolved compound was added to 95 μL of complete medium, resulting in a 20-fold dilution of the compound. Finally, 10 μL of the compound diluted in complete culture medium was added to each well of the cell suspension, resulting in nine concentration points representing a 5-fold serial dilution starting from 50 μM. A blank control containing 0.5% DMSO was included. The cells were incubated at 37°C in a 5% CO2 cell culture incubator for 5 days. On the sixth day, 50 μL of the compound was added to each well of the 96-well cell culture plate. The luminescent cell activity assay reagent (Promega, G7573) was incubated at room temperature for 10 minutes. The luminescence signal values ​​were then read using a multi-functional microplate reader (PerkinElmer, EnVision2015). The IC50 of the compound's inhibitory activity was calculated using Graphpad Prism software. 50 The values ​​are shown in Table I.

[0157] Table I shows the IC50 of the disclosed compounds in inhibiting NCI-H929 cell proliferation. 50 value

[0158] 1 0.02

[0159] Conclusion: Compound 1 disclosed herein exhibits excellent inhibitory activity against the proliferation of NCI-H929 cells.

[0160] Test Example 2: Efficacy Test

[0161] 1. Experimental Objective

[0162] The effects of compound 1 and control CC-92480 on inhibiting the growth of human multiple myeloma cell NCI-H929 xenografts in CB-17SCID mice were evaluated.

[0163] 2. Experimental reagents

[0164] Compound of Example 1;

[0165] Comparative Example CC-92480 (see Compound 2 in WO2019014100A1, synthesized according to the method disclosed therein)

[0166]

[0167] The compound of Example 1 and Comparative Example CC-92480 were formulated with 5% DMSO + 20% PEG400 + 70% (10% TPGS) + 5% (1% HPMC K100LV).

[0168] 3. Experimental Methods and Experimental Materials

[0169] 3.1. Experimental Animals and Feeding Conditions

[0170] Thirty female CB-17 SCID mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (Certificate No.: 20170011006049, SCXK (Hu) 2017-0011). The body weight was approximately 19 g when purchased. The mice were housed at 5 animals per cage, with a 12 / 12-hour light / dark cycle, constant temperature of 23±1°C, humidity of 50-60%, and free access to food and water.

[0171] 3.2. Animal Grouping:

[0172] After acclimatization feeding, CB-17 SCID mice were grouped as follows:

[0173]

[0174]

[0175] Note: qd means administration once a day; i.g means intragastric administration.

[0176] 3.3. Experimental Method:

[0177] NCI-H929 cells in logarithmic growth phase at 5×10 6 cells / mouse / 100 μL (containing 50 μL Matrigel) were inoculated subcutaneously into the right flank of 30 female CB-17 SCID mice. After 10 days, when the tumor volume of tumor-bearing mice reached approximately 200 mm 3 , the mice were randomly divided into 3 groups according to tumor volume and body weight: vehicle control group, CC-92480-1 mpk, compound of Example 1-1 mpk, with 7 mice in each group. The day of grouping was defined as Day 0 (D0), and intragastric administration was started once a day for a total of 11 days (Table 2). Tumor volume of tumor-bearing mice was measured with a vernier caliper and body weight was measured with a balance twice a week, and the data were recorded. When the tumor volume reached 2000 mm 3 or most tumors ulcerated or the body weight decreased by 20%, the tumor-bearing animals were euthanized as the experimental endpoint.

[0178] 3.4 Data Statistics

[0179] All data were plotted and statistically analyzed using Excel and GraphPad Prism 5 software.

[0180] The formula for calculating tumor volume (V) is: V = 1 / 2 × a × b 2 , where a and b represent length and width, respectively.

[0181] The relative tumor proliferation rate T / C (%) = (T-T0) / (C-C0) × 100 (%), where: T and C are the tumor volumes of the treatment group and the control group at the end of the experiment; T0 and C0 are the tumor volumes at the beginning of the experiment.

[0182] Tumor inhibition rate TGI (%) = 1 - T / C (%). When TGI (%) exceeds 100%, the specific value will not be displayed, but only >100% will be used.

[0183] Tumor regression (%) = [(T0-T) / T0] × 100 (%).

[0184] 4. Results

[0185] The efficacy data of compound CC-92480 against NCI-H929 xenografts in CB-17SCID mice in Example 1 are shown in Table II below. Figure 9 .

[0186] The effects of compound CC-92480 and control CC-92480 on body weight of CB-17SCID mice in Example 1 are shown below. Figure 10 .

[0187] Table II. Efficacy of the disclosed compounds against NCI-H929 xenografts in CB-17SCID mice.

[0188]

[0189] Note: qd means once a day; po means oral administration.

[0190] 5. Conclusion

[0191] In Example 1, the compound was administered once daily, starting 10 days after tumor cell transplantation. Significant tumor regression occurred after 11 days of administration. The calculated tumor inhibition rate was >100%, and the tumor regression rate was 88%, showing a statistically significant difference compared to the same dose of CC-92480 at the experimental endpoint (p < 0.05). Furthermore, the administration had no effect on mouse body weight. Under the same conditions, the tumor regression rate of the control group CC-92480 was 34%.

[0192] Test Example 3: Pharmacokinetic Evaluation

[0193] 1. Summary

[0194] Using mice as test animals, the LC / MS / MS method was used to determine the drug concentration in plasma at different time points after intragastric administration of the compound of Example 1 and the control compound CC-92480 to mice. The pharmacokinetic behavior of the compounds of the present disclosure in mice was studied, and their pharmacokinetic characteristics were evaluated.

[0195] 2. Test Protocol

[0196] 2.1 Test Drugs

[0197] The compound of Example 1 and the control compound CC-92480.

[0198] 2.2 Test Animals

[0199] Eighteen female mice were purchased from Vital River Laboratory Animal Co., Ltd., with the animal production license number: SCXK (Shanghai) 2017-0005.

[0200] 2.3 Drug Preparation

[0201] Weigh the compound of Example 1, add 5% by volume of DMSO and 5% Tween 80 (Shanghai Titan Technology Co., Ltd.) to dissolve it, then add 90% physiological saline to prepare a clear solution of 0.1 mg / mL.

[0202] Weigh the control compound CC-92480, add 5% by volume of DMSO and 5% Tween 80 (Shanghai Titan Technology Co., Ltd.) to dissolve it, then add 90% physiological saline to prepare a clear solution of 0.1 mg / mL.

[0203] 2.4 Administration

[0204] Nine mice were intragastrically administered with the compound of Example 1 at a dose of 2 mg / kg, and the administration volume was 0.2 mL / 10 g.

[0205] Nine mice were intragastrically administered with the control compound CC-92480 at a dose of 2 mg / kg, and the administration volume was 0.2 mL / 10 g.

[0206] 3. Procedure

[0207] After intragastric administration of the compound of Example 1 and the control compound CC-92480 to mice, 0.2 mL of blood was collected 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 (3 animals at each time point), and placed in EDTA-K2 anticoagulant tubes, followed by centrifugation at 10000 rpm for 1 minute at 4°C. Plasma was separated within 1 hour and stored at -20°C for testing. The process from blood collection to centrifugation was carried out under ice bath conditions.

[0208] Determination of the content of the analyte compound in mouse plasma after administration of different drug concentrations: 25 μL of mouse plasma was collected at each time point after drug administration, and 50 μL (100 ng / mL) of internal standard solution camptothecin (China National Institutes for Biological Products Control) and 175 μL of acetonitrile were added. The mixture was vortexed for 5 minutes and centrifuged for 10 minutes (3700 rpm). 1 μL of the supernatant from the plasma sample was analyzed by LC / MS / MS (API4000 triple quadrupole tandem mass spectrometer, Applied Biosystems, USA; Shimadzu LC-30AD ultra-high performance liquid chromatography system, Shimadzu Corporation, Japan).

[0209] 4. Pharmacokinetic Parameter Results

[0210] The pharmacokinetic parameters of the compounds disclosed herein are shown in Table III below.

[0211] Table III. Pharmacokinetic parameters of the compounds disclosed herein

[0212]

[0213] Conclusion: The compound disclosed in this study exhibits good pharmacokinetic absorption and has advantages in pharmacokinetic properties.

[0214] Test Example 4: Evaluation of the plasma stability of the disclosed compounds

[0215] 1. Abstract

[0216] The stability of the compound of Example 1 and CC-92480 and the control example CC-92480 in monkey frozen plasma at 37°C for 0, 15, 30, 60, 120, 180 and 240 minutes was determined by LC-MS / MS.

[0217] 2. Test Plan

[0218] 2.1 Test Drugs

[0219] Compound of Example 1 and Control Example CC-92480.

[0220] 2.2 Test plasma

[0221] Monkey plasma was purchased from Shanghai Medicilon Biopharmaceutical Co., Ltd.

[0222] 2.3 Preparation of Compound Solutions

[0223] Weigh a certain amount of the compound from Example 1 and add DMSO to prepare a 30 mM stock solution. Take a certain volume of the stock solution and dilute it with DMSO to prepare a 1600 μM solution I. Then take a certain volume of the 1600 μM solution I and dilute it with 50% methanol to prepare a 16 μM working solution II. Prepare the 30 mM stock solution, 1600 μM solution I', and 16 μM working solution II' of CC-92480 using the above method.

[0224] 2.4 Sample incubation

[0225] Take 5 μL of the working solution of 16 μM of the compound from Example 1 and the control example CC-92480, and add them to 75 μL of plasma respectively to bring the final concentration of the compound to 1 μM. Incubate the samples in a 37°C water bath for 0, 15, 30, 60, 90, 120, and 180 minutes. After incubation, add 240 μL of acetonitrile containing the internal standard, then shake at 800 rpm for 10 minutes, centrifuge at 3700 rpm at 4°C for 20 minutes, and analyze the supernatant using LC-MS with an injection volume of 2 μL.

[0226] 3. Results

[0227] The conversion of the disclosed compounds in monkey plasma is shown in Table IV below.

[0228] Table IV. Stability data of the disclosed compounds in monkey plasma.

[0229]

[0230]

[0231] Conclusion: The disclosed compound has a stability advantage in monkey plasma.

[0232] Example 2: Preparation of p-toluenesulfonate crystal form a of the compound shown in formula (I)

[0233] 10.08 mg of the compound shown in formula (I) was added to 0.4 mL of acetonitrile, heated, and then 8.96 μL of 2 M p-toluenesulfonic acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis identified the product as crystal form a, and the XRPD pattern is shown below. Figure 1 As shown in Table 1, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peaks have peak values ​​of 62.18℃ and 131.19℃. The TGA spectrum shows a weight loss of 2.26% between 30℃ and 210℃. Ion chromatography analysis indicates that the salt ratio of the compound shown in formula (I) to p-toluenesulfonic acid is 1:1.

[0234] Table 1

[0235]

[0236] Example 3: Preparation of p-toluenesulfonate crystal form b of the compound shown in formula (I)

[0237] 9.37 mg of the compound shown in formula (I) was added to 0.4 mL of tetrahydrofuran, heated, and then 8.96 μL of 2 M p-toluenesulfonic acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis identified the product as crystal form b. The XRPD spectrum is shown below. Figure 2 As shown in Table 2, the positions of its characteristic peaks are as follows. The DSC spectrum shows endothermic peaks at 76.87℃, 89.87℃, and 132.56℃. The TGA spectrum shows a weight loss of 3.67% between 30℃ and 210℃.

[0238] Table 2

[0239]

[0240] Example 4: Preparation of methanesulfonate crystal form a of the compound shown in formula (I)

[0241] 8.68 mg of the compound shown in formula (I) was added to 0.4 mL of acetonitrile, heated, and then 8.96 μL of 2 M methanesulfonic acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis showed that the product was of crystal form a, and the XRPD spectrum is shown below. Figure 3 As shown in Table 3, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 206.79℃. The TGA spectrum shows a weight loss of 0.01% from 30℃ to 190℃. Ion chromatography analysis shows that the salt ratio of the compound shown in formula (I) to methanesulfonic acid is 1:1.

[0242] Table 3

[0243]

[0244] Example 5: Preparation of methanesulfonate crystal form b of the compound shown in formula (I)

[0245] 9.76 mg of the compound shown in formula (I) was added to 0.4 mL of tetrahydrofuran, heated, and then 8.96 μL of 2 M methanesulfonic acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis showed that the product was crystal form b, and the XRPD pattern is shown below. Figure 4 As shown in Table 4, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 208.96℃. The TGA spectrum shows that the weight loss is 1.20% from 30℃ to 195℃.

[0246] Table 4

[0247]

[0248]

[0249] Example 6: Preparation of methanesulfonate crystal form c of the compound shown in formula (I)

[0250] The methanesulfonate b crystal form of the compound shown in formula (I) was placed in a DVS to obtain the product. X-ray powder diffraction analysis showed that the product was crystal form c, and the XRPD spectrum is shown below. Figure 5 As shown in Table 5, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peaks have peak values ​​of 82.64℃ and 206.30℃.

[0251] Table 5

[0252]

[0253] Example 7: Preparation of phosphate crystal form a of the compound shown in formula (I)

[0254] 10.01 mg of the compound shown in formula (I) was added to 0.4 mL of acetonitrile, heated, and then 8.96 μL of 2 M phosphoric acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis identified the product as crystal form a, and the XRPD pattern is shown below. Figure 6 As shown in Table 6, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peaks have peak values ​​of 208.82℃ and 220.84℃. The TGA spectrum shows a weight loss of 0.78% between 30℃ and 220℃. Ion chromatography analysis indicates that the salt ratio of the compound shown in formula (I) to phosphoric acid is 1:1.

[0255] Table 6

[0256]

[0257]

[0258] Example 8: Preparation of hydrobromide crystal form a of the compound shown in formula (I)

[0259] 9.85 mg of the compound shown in formula (I) was added to 0.4 mL of acetonitrile, heated, and then 8.96 μL of 2 M hydrobromic acid solution was added. The mixture was cooled, and a solid precipitated. The solid was centrifuged and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis identified the product as crystal form a, and the XRPD pattern is shown below. Figure 7 As shown in Table 7, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 195.15℃. The TGA spectrum shows a weight loss of 5.66% from 30℃ to 200℃. Ion chromatography analysis shows that the salt ratio of the compound shown in formula (I) to hydrobromic acid is 1:1.

[0260] Table 7

[0261]

[0262] Example 9: Preparation of crystal form a of the hydrochloride salt of the compound shown in formula (I)

[0263] 10.37 mg of the compound shown in formula (I) was added to 0.4 mL of acetonitrile and 8.96 μL of 2 M hydrochloric acid solution. The mixture was heated, cooled to precipitate a solid, centrifuged, and dried under vacuum at 45 °C to obtain the product. X-ray powder diffraction analysis identified the product as crystal form a, and the XRPD pattern is shown below. Figure 8 As shown in Table 8, the positions of its characteristic peaks are as follows. The DSC spectrum shows that the endothermic peak has a peak value of 159.52℃. The TGA spectrum shows that the weight loss is 5.92% from 30℃ to 205℃. Ion chromatography analysis shows that the salt formation ratio of the compound shown in formula (I) to hydrochloric acid is 1:1.

[0264] Table 8

[0265]

[0266]

[0267] Example 10: Hygroscopicity Study

[0268] Using Surface Measurement Systems advantage 2, at 25°C and humidity starting from 50%, the humidity range was investigated from 0% to 95%, with a step size of 10%. The judgment criterion was that the mass change dM / dT for each gradient was less than 0.002%, TMAX 360 min, and two cycles were performed.

[0269] Table 9

[0270]

[0271] Example 11: Stability of Influencing Factors

[0272] The samples of hydrobromide a crystal form, methanesulfonate a crystal form, methanesulfonate b crystal form, and hydrochloride a crystal form were laid out in the open and their stability was investigated under light (4500 Lux), high temperature (40℃, 60℃), and high humidity (RH 75%, RH 92.5%) conditions. The sampling period was 30 days.

[0273] Table 10

[0274]

[0275]

[0276]

[0277] Conclusion: The hydrobromide a crystal form and the hydrochloride a crystal form exhibit good physicochemical stability under high temperature (40℃ and 60℃) and high humidity (75% and 92.5%) conditions, although their purity slightly decreases under light exposure at 40℃ and 60℃. The methanesulfonate b crystal form also exhibits good physicochemical stability under high temperature (40℃ and 60℃) and high humidity (92.5%) conditions.

[0278] Example 12: Long-term / accelerated stability

[0279] The stability of hydrobromide a crystal form, methanesulfonate a crystal form, methanesulfonate b crystal form, and hydrochloride a crystal form was investigated under conditions of 25°C, 60% RH, and 40°C, 75% RH, respectively.

[0280]

[0281]

[0282]

[0283] Long-term / accelerated stability tests showed that the hydrobromide a crystal form and the hydrochloride a crystal form exhibited good physical and chemical stability after 6 months of storage. The methanesulfonate b crystal form showed good physical stability after 6 months of storage, although its chemical purity decreased slightly.

Claims

1. A pharmaceutically acceptable salt of the compound shown in formula (I), wherein the pharmaceutically acceptable salt is selected from p-toluenesulfonate, methanesulfonate, phosphate, hydrochloride and hydrobromide. Formula I.

2. The medicinal salt according to claim 1, characterized in that, The chemical ratio of the compound shown in formula (I) to the acid molecule is 5:1 to 1:

5.

3. The medicinal salt according to claim 1, characterized in that, The chemical ratio of the compound shown in formula (I) to the acid molecule is 3:1, 2:1, 1:1, 1:2, 1:3, 1:4 or 1:

5.

4. The medicinal salt according to claim 1, characterized in that, The chemical ratio of the compound shown in formula (I) to the acid molecule is 2:1, 1:1 or 1:

2.

5. The α-crystal form of the compound p-toluenesulfonate shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 5.787, 6.961, 9.961, 15.756, 21.717, 23.539, and 26.

272.

6. The a-crystal form of the compound of formula (I) according to claim 5, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 1.

7. The b-crystal form of the p-toluenesulfonate of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 5.823, 11.447, 12.455, 13.772, 17.807, 22.501, and 23.

072.

8. The b-crystal form of the compound of formula (I) according to claim 7, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 2.

9. The α-crystal form of the methanesulfonate of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.616, 11.915, 17.525, 19.197, 22.233, 24.561, and 25.

679.

10. The α-crystal form of the methanesulfonate of formula (I) according to claim 9, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.616, 11.915, 17.525, 19.197, 19.969, 22.233, 24.124, 24.561, 25.679, and 27.

524.

11. The α-crystal form of the methanesulfonate of formula (I) according to claim 9, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.616, 11.915, 17.525, 19.197, 19.969, 22.233, 24.124, 24.561, 25.679, 27.524, 28.725, 31.000, and 34.

434.

12. The α-crystal form of the methanesulfonate of formula (I) according to claim 9, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 3.

13. The b-crystal form of the methanesulfonate of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, and 24.

655.

14. The b-crystal form of the methanesulfonate of formula (I) according to claim 13, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, 24.655, 26.357, 29.109, and 32.

893.

15. The b-crystal form of the methanesulfonate of formula (I) according to claim 13, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.690, 11.263, 17.505, 19.175, 20.988, 23.275, 24.655, 26.357, 29.109, 32.893, and 35.

688.

16. The b-crystal form of the methanesulfonate of formula (I) according to claim 13, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 4.

17. The c-crystal form of the methanesulfonate of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 11.191, 16.225, 17.493, 19.622, 22.683, 24.308, and 26.

271.

18. The c-crystal form of the methanesulfonate of formula (I) according to claim 17, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 11.191, 15.027, 16.225, 17.493, 19.622, 22.683, 24.308, 25.113, and 26.

271.

19. The c-crystal form of the methanesulfonate of formula (I) according to claim 17, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 5.

20. The α-crystal form of the phosphate of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.525, 12.180, 14.251, 17.952, 20.210, 22.157, and 27.

662.

21. The α-crystal form of the phosphate of formula (I) according to claim 20, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.525, 12.180, 14.251, 17.952, 19.282, 20.210, 22.157, 24.560, 25.821, and 27.

662.

22. The α-crystal form of the phosphate of formula (I) according to claim 20, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 8.525, 12.180, 14.251, 16.158, 17.952, 19.282, 20.210, 22.157, 24.560, 25.404, 25.821, 27.662, and 28.

781.

23. The α-crystal form of the phosphate of formula (I) according to claim 20, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 6.

24. The α-crystal form of the hydrobromide of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 9.405, 10.749, 17.979, 19.610, 20.887, 22.693, and 26.

133.

25. The α-crystal form of the hydrobromide of formula (I) according to claim 24, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 9.405, 10.749, 12.992, 15.520, 17.979, 19.610, 20.887, 22.693, 25.458, and 26.

133.

26. The α-crystal form of the hydrobromide of formula (I) according to claim 24, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 9.405, 10.749, 12.992, 15.520, 17.979, 19.610, 20.887, 22.693, 25.458, 26.133, 27.099, 29.346, and 31.

679.

27. The α-crystal form of the hydrobromide of formula (I) according to claim 24, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 7.

28. The α-crystal form of the hydrochloride salt of the compound shown in formula (I), Formula I, in, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 7.019, 9.760, 10.683, 17.988, 21.349, 23.050, and 26.

039.

29. The α-crystal form of the hydrochloride salt of formula (I) according to claim 28, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 7.019, 9.760, 10.683, 13.645, 16.287, 17.988, 19.646, 21.349, 23.050, and 26.

039.

30. The α-crystal form of the hydrochloride salt of formula (I) according to claim 28, wherein, The X-ray powder diffraction pattern, expressed as a diffraction angle 2θ, shows characteristic peaks at 7.019, 9.760, 10.683, 12.844, 13.645, 16.287, 17.988, 19.646, 21.349, 23.050, and 26.

039.

31. The α-crystal form of the hydrochloride salt of formula (I) according to claim 28, wherein, The X-ray powder diffraction pattern expressed in terms of the diffraction angle 2θ is shown in Figure 8.

32. The crystal form according to any one of claims 5-31, characterized in that, The error range of the 2θ angle is ±0.

20.

33. A pharmaceutical composition comprising a pharmaceutically acceptable salt as described in any one of claims 1-4 or a crystal form as described in any one of claims 5-32, and optionally a pharmaceutically acceptable carrier, diluent, or excipient.

34. A method for preparing a pharmaceutical composition, comprising the step of mixing a pharmaceutically acceptable salt according to any one of claims 1-4 or a crystal form according to any one of claims 5-32 with a pharmaceutically acceptable carrier, diluent or excipient.

35. Use of the pharmaceutically acceptable salt of any one of claims 1-4, or the crystal form of any one of claims 5-32, or the pharmaceutical composition of claim 33, or the composition prepared by the method of claim 34, in the preparation of a medicament for treating and / or preventing cancer, wherein the cancer is selected from multiple myeloma.

36. The use according to claim 35, wherein the multiple myeloma is relapsed, refractory, or resistant.

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