A pharmaceutically acceptable salt crystal form of an endothelin A (ETA) receptor antagonist compound and a preparation method thereof

Abstract

The present invention belongs to the technical field of chemical medicines and provides a pharmaceutically acceptable salt crystal form of an endothelin A (ETA) receptor antagonist compound and a preparation method thereof.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical medicines and provides a pharmaceutically usable salt crystal form of an endothelin A (ETA) receptor antagonist compound, a preparation method and an application thereof. Background Art

[0002] Atrasentan (CAS: 173937-91-2) is a potent and selective endothelin A (ETA) receptor antagonist with the following structural formula: Atrasentan hydrochloride (CAS: 195733-43-8) is used clinically and has the following structure: It was previously evaluated in clinical trials for the treatment of prostate cancer and is currently being evaluated in clinical trials for the treatment of chronic kidney disease associated with type 2 diabetes. It has also been shown to reduce albuminuria in patients with diabetic nephropathy. Currently, there is no marketed product. Summary of the Invention

[0003] The applicant obtained the following compound A in the early stage of research:

[0004]

[0005] In one aspect, the present invention provides a crystalline form of a compound represented by (I) and a pharmaceutically acceptable salt thereof.

[0006]

[0007] Wherein: the M is selected from oxalic acid, wherein n=1; x=1.

[0008] As a preferred embodiment of the present invention, the crystal form is oxalate crystal form A, and the X-ray diffraction pattern shows characteristic peaks at 4.96°, 5.32°, 6.44°, 7.44°, and 13.33° in terms of 2θ angle, with an error of ±0.2°.

[0009] As a preferred embodiment of the present invention, the crystal form is oxalate crystal form A, and the X-ray diffraction pattern shows characteristic peaks at 4.96°, 5.32°, 6.44°, 7.44°, 13.33°, 19.41°, 19.81°, 20.22°, 20.89°, and 22.48° in terms of 2θ angle, with an error of ±0.2°.

[0010] As a preferred embodiment of the present invention, the X-ray powder diffraction spectrum of the oxalate crystal form A is shown in the attached Figure 1 or attached Figure 2 shown.

[0011] As a preferred embodiment of the present invention, the differential thermal analysis spectrum of the oxalate crystal form A has an endothermic peak at 76.7±5°C. The differential thermal analysis spectrum of the oxalate crystal form A is as follows: Figure 3 shown.

[0012] As a preferred embodiment of the present invention, the crystal form is oxalate crystal form B, and the X-ray diffraction pattern shows characteristic peaks at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 13.25°, 14.56°, 18.05°, 19.02°, and 21.65° in terms of 2θ angle, with an error of ±0.2°.

[0013] As a preferred embodiment of the present invention, the crystal form is oxalate crystal form B, and the X-ray diffraction pattern is represented by 2θ angles at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 10.40°, 12.76°, 13.25°, 14.56°, 15.75°, 16.19°, 18.05°, 19.02°, 20.85°, 21.65°, 22.38°, 22.76°, and 25.21°, with an error of ±0.2°.

[0014] As a preferred embodiment of the present invention, the X-ray powder diffraction spectrum of the oxalate crystal form B is as shown in the attached Figure 4 or attached Figure 5 shown.

[0015] As a preferred embodiment of the present invention, the differential thermal analysis spectrum of the oxalate crystal form B has an endothermic peak at 69.4±5°C. The differential thermal analysis spectrum of the oxalate crystal form B is as follows: Figure 6 shown.

[0016] As a preferred embodiment of the present invention, the crystal form is oxalate crystal form C, and the X-ray diffraction pattern shows characteristic peaks at 5.11°, 6.94°, 8.84°, and 13.16° in terms of 2θ angle, with an error of ±0.2°.

[0017] As a preferred embodiment of the present invention, the X-ray powder diffraction spectrum of the oxalate crystal form C is as shown in the attached Figure 7 or attached Figure 8 shown.

[0018] As a preferred embodiment of the present invention, the differential thermal analysis spectrum of the oxalate crystal form C has an endothermic peak at 63.3±5°C. The differential thermal analysis spectrum of the oxalate crystal form C is as follows: Figure 9 shown.

[0019] The present invention further provides a crystalline form of the compound shown in (I) and a pharmaceutically acceptable salt thereof,

[0020]

[0021] Wherein: the M is selected from phosphoric acid, wherein n=3; x=5.

[0022] As a preferred embodiment of the present invention, the crystal form is phosphate crystal form A, and the X-ray diffraction pattern shows characteristic peaks at 5.07°, 7.05°, 8.63°, and 13.25° in terms of 2θ angle, with an error of ±0.2°.

[0023] As a preferred embodiment of the present invention, the X-ray powder diffraction spectrum of the phosphate crystal form A is shown in the attached Figure 10 or attached Figure 11 shown.

[0024] The present invention further provides a pharmaceutical composition, characterized in that the pharmaceutical composition contains a therapeutically effective amount of the compound shown in one crystalline form of (I) and its pharmaceutically acceptable salt, and one or more pharmaceutically acceptable carriers.

[0025] The present invention further provides the use of the compound represented by (I) in a crystalline form and a pharmaceutically acceptable salt thereof in the preparation of drugs for treating and / or preventing diseases related to endothelin A (ETA) receptor antagonism.

[0026] As a preferred embodiment of the present invention, the diseases include chronic kidney disease, IgA, FSGS, Alport and hypertension.

[0027] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects on the host or patient. Representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, viscosity increasers, transdermal enhancers, etc. Their preparation is well known to those skilled in the field of cosmetics or topical medicine. For additional information on carriers, reference can be made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is the X-ray diffraction pattern of the oxalate crystal form A obtained in Example 15 of the present invention.

[0029] Figure 2 This is the X-ray diffraction pattern of the oxalate crystal form A obtained in Example 16 of the present invention.

[0030] Figure 3 This is the DSC spectrum of the oxalate crystal form A obtained in Example 15 of the present invention.

[0031] Figure 4 This is the X-ray diffraction pattern of the oxalate crystal form B obtained in Example 17 of the present invention.

[0032] Figure 5 This is the X-ray diffraction pattern of the oxalate crystal form B obtained in Example 18 of the present invention.

[0033] Figure 6 This is the DSC spectrum of the oxalate crystal form B obtained in Example 17 of the present invention.

[0034] Figure 7 This is the X-ray diffraction pattern of the oxalate crystal form C obtained in Example 19 of the present invention.

[0035] Figure 8 This is the X-ray diffraction pattern of the oxalate crystal form C obtained in Example 20 of the present invention.

[0036] Figure 9 This is the DSC spectrum of the oxalate crystal form C obtained in Example 19 of the present invention

[0037] Figure 10 This is the X-ray diffraction pattern of the phosphate crystal form A obtained in Example 21 of the present invention.

[0038] Figure 11 This is the X-ray diffraction pattern of the phosphate crystal form A obtained in Example 22 of the present invention.

[0039] Figure 12 This is the NMR spectrum of the hydrochloride obtained in Example 23 of the present invention

[0040] Figure 13 This is the NMR spectrum of the sulfate obtained in Example 24 of the present invention DETAILED DESCRIPTION

[0041] The present invention will be further described in detail below with reference to examples, but the embodiments of the present invention are not limited thereto.

[0042] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-III NMR spectrometer, using deuterated dimethyl sulfoxide (DMSO-d6) and deuterated chloroform (CDCl3) as solvents and tetramethylsilane (TMS) as the internal standard.

[0043] MS was measured using an ISQ EC mass spectrometer (manufacturer: Thermo, model: ISQ EC).

[0044] High performance liquid chromatography (HPLC) analysis was performed using a Thermo U3000 HPLC DAD instrument.

[0045] The CombiFlash rapid preparation instrument used CombiFlash Rf+LUMEN (TELEDYNE ISCO).

[0046] Thin layer chromatography silica gel plate using Yantai Yinlong HSGF 254 or GF 254 Silica gel plates, the specifications of silica gel plates used in thin layer chromatography (TLC) are 0.17mm~0.23mm, and the specifications used for thin layer chromatography separation and purification products are 0.4mm~0.5mm.

[0047] Silica gel column chromatography generally uses Rushan Shangbang silica gel 100-200 mesh silica gel as the carrier.

[0048] The present invention relates to the following reagents: DMF (N,N-dimethylformamide), KI (potassium iodide), Cs2CO3 (cesium carbonate), DCM (dichloromethane), n-hexane, EA (ethyl acetate), Py (pyridine), CF3COOH (trifluoroacetic acid), IPA (isopropyl alcohol), isopropyl ether, and acetone.

[0049] Unless otherwise specified, the polymorphs of the present invention were detected using the following equipment and conditions: X-ray powder diffraction (XRPD). XRPD patterns were collected on an X-ray powder diffraction analyzer produced by PANalytacal. The scanning parameters are as follows:

[0050]

[0051] Differential scanning calorimetry (DSC) was performed on a German NETZSCH DSC 200F3 differential scanning calorimeter. The test parameters are as follows:

[0052]

[0053] Example 1

[0054] 1-[(Ethoxycarbonyl)oxy]methyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0055]

[0056] Step A: Synthesis of 1-[(ethoxycarbonyl)oxy]methyl(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0057]

[0058] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1, 3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), chloromethyl ethyl carbonate (270 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 2 h.

[0059] After the reaction, the reaction solution was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (100 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated and evaporated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to give 480 mg of colorless oily product [(ethoxycarbonyl)oxy]methyl (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 79.9%).

[0060] LC-MS: RT = 2.20 min, [M+H] + =613.42.

[0061] 1H NMR (400MHz, DMSO) δ7.23 (d, J=8.6Hz, 2H), 7.04 (d, J=1.2Hz, 1H), 6.89 (d, J=8.7Hz, 2H), 6.84-6.76 (m, 2H), 5.98 ( d, J=4.6Hz, 2H), 5.58 (q, J=6.2Hz, 2H), 4.12 (q, J=7.1Hz, 2H), 3.78-3.72 (m, 1H), 3.72 (s, 3H), 3.49 (dd, J=11.2, 5. 3Hz, 1H), 3.26-3.20 (m, 3H), 3.16 (s, 1H), 3.02-2.90 (m, 3H), 2.86-2.80 (m, 1H), 2.71 (d, J=13.8Hz, 1H), 1.33 (dd, J=14.6, 7.0Hz, 2H), 1.27-1.09 (m, 7H), 0.96 (dd, J=14.7, 7.3Hz, 2H), 0.81 (t, J=7.3Hz, 3H), 0.71 (t, J=7.3Hz, 3H).

[0062] Example 2

[0063] 1-[(Ethoxycarbonyl)oxy]ethyl (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0064]

[0065] Step A: Synthesis of 1-[(ethoxycarbonyl)oxy]ethyl (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0066]

[0067] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1, 3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), 1-chloroethyl ethyl carbonate (298 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 2 h.

[0068] After the reaction, the reaction solution was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (100 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated and evaporated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to give 442 mg of colorless oily product 1-[(ethoxycarbonyl)oxy]methyl (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 66.5%).

[0069] LC-MS: RT = 2.21 min, [M+H] + =627.40. 1 H NMR (400MHz, DMSO) δ7.27-7.17(m, 2H), 7.09-6.98(m, 1H), 6.92-6.85(m, 2H), 6.85-6.74(m, 2H), 6.60-6. 52 (m, 1H), 5.98 (d, J=5.3Hz, 2H), 4.11 (dq, J=11.1, 7.1Hz, 2H), 3.67-3.73 (m, 4H), 3.57-3.44 (m, 1H), 3.30 -3.20 (m, 3H), 3.19-3.10 (m, 1H), 3.01-2.85 (m, 3H), 2.77 (ddd, J=9.4, 6.9, 2.6Hz, 1H), 2.70 (d, J=13.9Hz, 1H), 1.39-1.22(m, 6H), 1.21-1.04(m, 6H), 1.01-0.89(m, 2H), 0.81(t, J=7.3Hz, 3H), 0.71(t, J=7.3Hz, 3H)

[0070] Example 3

[0071] 1-[(Isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0072]

[0073] Step A: Synthesis of 1-[(isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0074]

[0075] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1, 3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), 1-chloroethyl isopropyl carbonate (325 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 2 h.

[0076] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to give 482 mg of colorless oily product 1-[(isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 76.9%).

[0077] LC-MS: RT = 2.29 min, [M+H] + =641.48. 1 H NMR (400MHz, DMSO) δ7.25 (t, J=9.0Hz, 2H), 7.07-7.04 (m, 1H), 6.93-6.89 (m, 2H), 6.85-6.78 (m, 2H), 6.59-6.54 (m, 1H), 5.99 (d, J=6.2Hz, 2H), 4.79-4.69 (m, 1H), 3.76-3.70 (m, 4H), 3.55-3.44 (m, 1H), 3.33-3.23 (m, 4H), 3.21 -3.13 (m, 1H), 3.02-2.90 (m, 3H), 2.80-2.75 (m, 1H) 2.71 (d, J = 13.9Hz, 1H), 1.39-1.33 (m, 1H), 1.32 (d, J = 5.4Hz , 2H), 1.26 (d, J=5.4Hz, 2H), 1.22-1.14 (m, 9H), 1.00-0.94 (m, 2H), 0.82 (t, J=7.3Hz, 3H), 0.72 (t, J=7.3Hz, 3H).

[0078] Example 4

[0079] 1-[(Methoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0080]

[0081] Step A: Synthesis of 1-[(methoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0082]

[0083] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), 1-chloroethyl methyl carbonate (270 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 2 h.

[0084] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to give 428 mg of colorless oily product 1-[(methoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 71.3%).

[0085] LC-MS: RT = 2.20 min, [M+H] + =613.42. 1H NMR (400MHz, DMSO) δ7.25 (dd, J=10.3, 8.7Hz, 2H), 7.08-7.04 (m, 1H), 6.91 (dd, J=8.7, 2.9 Hz, 2H), 6.86-6.78 (m, 2H), 6.60-6.55 (m, 1H), 5.99 (d, J=5.1Hz, 2H), 3.76-3.69 (m, 6H), 3. 56-3.44(m, 1H), 3.33-3.16(m, 5H), 3.00-2.94(m, 3H), 2.81-2.76(m, 1H), 2.71(d, J=13.7H z, 1H), 1.39-1.14 (m, 9H), 1.02-0.93 (m, 2H), 0.82 (t, J=7.3Hz, 3H), 0.72 (t, J=7.3Hz, 3H).

[0086] Example 5

[0087] 1-(((2-(Methylamino)ethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0088]

[0089] Step A: Synthesis of tert-butyl (2-(((1-chloroethoxy)carbonyl)oxy)ethyl)(methyl)carbamate

[0090]

[0091] Under ice bath, 2-(N-Boc-N-methylamino)ethanol (500 mg, 2.85 mmol) and pyridine (248 mg, 3.14 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (248 mg, 3.14 mmol) was added dropwise under ice bath. After the addition was completed, the temperature was raised to room temperature and the reaction was carried out for 1 hour.

[0092] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), and the organic layer was washed twice with 1 M / L hydrochloric acid (20 ml), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure to give 725 mg of a colorless oily product (tert-butyl 2-(((1-chloroethoxy)carbonyl)oxy)ethyl)(methyl)carbamate (yield: 90.6%).

[0093] Step B: Synthesis of 2,2,5-trimethyl-4,9-dioxo-3,8,10-trioxa-5-aza-11-ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0094]

[0095] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1, 3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), tert-butyl (2-(((1-chloroethoxy)carbonyl)oxy)ethyl)(methyl)carbamate (550 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 4 h.

[0096] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution. The mixture was extracted with dichloromethane (20 ml x 3). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to obtain 625 mg of a colorless oily product, 2,2,5-trimethyl-4,9-dioxo-3,8,10-trioxa-5-aza-11-ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 84.4%). LC-MS: RT = 2.34 min, [M+H] + =756.49.

[0097] Step C: Synthesis of 1-(((2-(methylamino)ethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0098]

[0099] At room temperature, 2,2,5-trimethyl-4,9-dioxo-3,8,10-trioxa-5-aza-11-ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (300 mg, 0.39 mmol) was dissolved in 10 ml of ethyl acetate solvent, and 2 ml of 4 M / L hydrochloric acid dioxane solution was added under ice bath, and the reaction was carried out at room temperature for 2 hours.

[0100] After the reaction, the reaction mixture was concentrated to obtain 260 mg of a yellow solid 1-(((2-(methylamino)ethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 101.6%).

[0101] LC-MS: RT = 1.82 min, [M+H] + =656.45. 1 H NMR (400MHz, DMSO) δ7.60 (m, 2H), 7.21 (m, 1H), 7.01 (d, J=8.1Hz, 2H), 6.90 (d, J=15.8Hz, 2H), 6.50 (m, 1H), 6.04 (s, 2H), 3.77 (s, 3H), 3.17 (d, J=6.4Hz , 3H), 3.08-2.90 (m, 4H), 2.52 (d, J=5.6Hz, 3H), 2.06 (d, J=4.1Hz, 3H), 1.9 0 (s, 6H), 1.33 (d, J = 34.4Hz, 4H), 1.23-1.10 (m, 7H), 0.83 (t, J = 7.0Hz, 6H).

[0102] Example 6

[0103] 1-((((S)-2,3-dihydroxypropoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0104]

[0105] Step A: 1-Chloroethyl (((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl)carbonate

[0106]

[0107] Under ice bath, (S)-(+)-1,2-isopropyl glycerol (315 mg, 2.38 mmol) and pyridine (225 mg, 2.85 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (408 mg, 2.85 mmol) was added dropwise under ice bath. After the addition was completed, the temperature was raised to room temperature and the reaction was carried out for 2 hours.

[0108] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to give 250 mg of colorless oily product 1-chloroethyl (((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl) carbonate (yield: 44.2%).

[0109] Step B: Synthesis of 1-(((((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0110]

[0111] Atrasentan (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (100 mg, 0.20 mmol), 1-chloroethyl (((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl) carbonate (72 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol) and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF at room temperature and the temperature was raised to 65°C for 4 hours.

[0112] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution. The mixture was extracted with dichloromethane (20 ml x 3). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to obtain 85 mg of colorless oily product, 1-(((((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 59.9%). LC-MS: RT = 2.21 min, [M+H] + =713.46.

[0113] Step C: Synthesis of 1-((((S)-2,3-dihydroxypropoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0114]

[0115] At room temperature, 1-(((((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (80 mg, 0.11 mmol) was dissolved in 10 ml of dichloromethane solvent, 2 ml of trifluoroacetic acid solution was added, and the reaction was carried out at room temperature for 0.5 hour.

[0116] After the reaction, the crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 2) to obtain 56 mg of a yellow solid 1-((((S)-2,3-dihydroxypropoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 74.1%).

[0117] LC-MS: RT = 2.06 min, [M+H] + =673.39. 1H NMR (400MHz, DMSO) δ7.32-7.17(m, 2H), 7.12-7.04(m, 1H), 6.93(s, 2H), 6.84(s, 2H), 6. 55 (s, 1H), 6.00 (d, J=5.2Hz, 2H), 4.18-4.06 (m, 2H), 3.90 (s, 1H), 3.73 (s, 3H), 3.64 (s, 2 H), 3.55-3.42(m, 2H), 3.18-3.07(m, 3H), 3.00(s, 4H), 2.66(s, 1H), 2.44-2.35(m, 2H), 2.32 (s, 1H), 1.31 (s, 3H), 1.18 (s, 3H), 0.99 (s, 3H), 0.81 (t, J=7.4Hz, 3H), 0.74 (s, 3H).

[0118] Example 7

[0119] 1-[(Cyclopropyloxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0120]

[0121] Step A: Synthesis of 1-[(cyclopropyloxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0122]

[0123] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1, 3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (500 mg, 0.98 mmol), 1-chloroethylcyclopropyl carbonate (240 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol) and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF and heated to 65 °C for 2 h.

[0124] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution. Extraction was performed with dichloromethane (100 ml x 3). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to obtain 412 mg of colorless oily product, 1-[(cyclopropyloxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 65.8 g). LC-MS: RT = 2.29 min, [M+H] + =639.40. 1 H NMR (400MHz, DMSO) δ7.25 (s, 2H), 7.06 (d, J=10.4Hz, 1H), 6.90 (m, 2H), 6.83 (m, 2H), 6.57 (m, 1H), 5.98 (s, 2H), 4.09 (s, 2H), 3.73 (m, 4H), 3.5 3-3.42 (m, 2H), 2.95 (m, 5H), 2.71 (d, J=13.2Hz, 3H), 1.98 (s, 1H), 1.38-1.02 (m, 9H), 0.99-0.87 (m, 2H), 0.81 (s, 2H), 0.71 (d, J=7.0Hz, 6H).

[0125] Example 8

[0126] 1-(((2-hydroxyethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0127]

[0128] Step A: Synthesis of 2-((tert-butyldimethylsilyl)oxy)ethyl(1-chloroethyl)carbonate

[0129]

[0130] Under ice bath, 2-tert-butyldimethylsilyloxyethanol (1000 mg, 5.67 mmol) and pyridine (739 mg, 9.36 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (900 mg, 6.24 mmol) was added dropwise under ice bath. After the addition was completed, the temperature was raised to room temperature and the reaction was carried out for 1 hour.

[0131] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to give 245 mg of colorless oily product 2-((tert-butyldimethylsilyl)oxy)ethyl (1-chloroethyl) carbonate (yield: 15.4%).

[0132] Step B: Synthesis of 2,2,3,3-tetramethyl-8-oxo-4,7,9-trioxa-3-sila-10-ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0133]

[0134] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (100 mg, 0.20 mmol), 2-((tert-butyldimethylsilyl)oxy)ethyl(1-chloroethyl)carbonate (85 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol) and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF and heated to 65 °C for 4 h.

[0135] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution. The mixture was extracted with dichloromethane (20 ml x 3). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to obtain 62 mg of colorless oily product, 2,2,3,3-tetramethyl-8-oxo-4,7,9-trioxa-3-sila-10-ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 41.0%). LC-MS: RT = 2.75 min, [M+H] + =757.50.

[0136] Step C: Synthesis of 1-(((2-hydroxyethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0137]

[0138] At room temperature, 2,2,3,3-tetramethyl-8-oxo-4,7,9-trioxa-3-sila-10-ethyl-(2R,3R,4SS)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (62 mg, 0.08 mmol) was dissolved in 10 ml of dichloromethane solvent, and 2 ml of 4 M / L hydrochloric acid dioxane solution was added under ice bath, and the reaction was carried out at room temperature for 2 hours.

[0139] After the reaction, the crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 2) to give 40 mg of a yellow solid 1-(((2-hydroxyethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 77.9%).

[0140] LC-MS: RT = 2.07 min, [M+H] + =643.44. 1 H NMR (400MHz, DMSO) δ7.71-7.38 (m, 2H), 7.19 (d, J=12.2Hz, 1H), 7.00 (d, J=6.8Hz, 2H), 6.93-6.79 (m, 2H), 6.48 (s, 1H), 6.03 (s, 2H), 4.02 (d, J=7.1Hz, 3H), 3.76 (m, 4H), 3.57-3.48 (m, 3H), 3.20-3.09 (m, 1H), 3.08-2.90 (m, 3H), 1.45-1.02 (m, 14H), 0.91-0.68 (m, 8H).

[0141] Example 9

[0142] 1-(((((R)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0143]

[0144] Step A: ((R)-1,4-dioxan-2-yl)methyl (1-chloroethyl) carbonate

[0145]

[0146] Under ice bath, (R)-(1,4-dioxane-2-yl)methanol (280 mg, 2.38 mmol) and pyridine (225 mg, 2.85 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (408 mg, 2.85 mmol) was added dropwise under ice bath. After the addition was completed, the temperature was raised to room temperature and the reaction was carried out for 2 hours.

[0147] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to give 212 mg of a colorless oily product ((R)-1,4-dioxan-2-yl)methyl (1-chloroethyl) carbonate (yield: 39.8%).

[0148] Step B: Synthesis of 1-(((((R)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0149]

[0150] At room temperature, atrasentan (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (100 mg, 0.20 mmol), ((R)-1,4-dioxan-2-yl)methyl(1-chloroethyl)carbonate (67.2 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol) and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF and heated to 65°C for 4 hours.

[0151] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to give 80 mg of colorless oily product 1-(((((R)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 59.9%).

[0152] LC-MS: RT = 2.16 min, [M+H] + =699.42. 1 H NMR (400MHz, DMSO) δ7.24 (dd, J=11.6, 8.6Hz, 2H), 7.06 (d, J=11.4Hz, 1H), 6.91 (dd, J=8.6, 2.7Hz, 2H), 6.85-6.75 (m, 2H) , 6.59-6.53 (m, 1H), 5.99 (d, J=6.0Hz, 2H), 4.06 (dd, J=9.5, 4.7Hz, 2H), 3.75-3.56 (m, 8H), 3.53 (d, J=9.9Hz, 1H), 3.50-3. 41 (m, 2H), 3.29-3.21 (m, 5H), 2.95 (dd, J=18.3, 10.6Hz, 3H), 2.79-2.73 (m, 1H), 2.69 (d, J=14.1Hz, 1H), 1.33 (t, J=8.5Hz, 3H), 1.29-1.21 (m, 3H), 1.17 (dd, J=14.8, 7.4Hz, 3H), 0.96 (d, J=4.7Hz, 2H), 0.81 (t, J=7.3Hz, 3H), 0.71 (t, J=7.3Hz, 3H).

[0153] Example 10

[0154] 1-((((oxetan-3-yl)oxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0155]

[0156] Step A: 1-Chloroethyloxetan-3-yl carbonate

[0157]

[0158] Under ice bath, oxetane-3-ol (176 mg, 2.38 mmol) and pyridine (225 mg, 2.85 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (408 mg, 2.85 mmol) was added dropwise under ice bath. After the addition was completed, the mixture was warmed to room temperature and reacted for 2 hours.

[0159] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure and evaporated to dryness. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to give 200 mg of colorless oily product 1-chloroethyloxetane-3-yl carbonate (yield: 46.7%).

[0160] Step B: Synthesis of 1-((((oxetan-3-yl)oxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0161]

[0162] At room temperature, atrasentan (2R, 3R, 4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (100 mg, 0.2 mmol), 1-chloroethyloxetane-3-yl carbonate (54 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol) and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF and heated to 65 °C for 4 h.

[0163] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), and the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to give 76 mg of colorless oily product 1-((((oxetan-3-yl)oxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 58.0%).

[0164] LC-MS: RT = 2.18 min, [M+H] + =655.39. 1 H NMR (400MHz, DMSO) δ7.24 (t, J=8.4Hz, 2H), 7.05 (d, J=12.1Hz, 1H), 6.94-6.86 (m, 2H), 6.85-6.71 (m, 2H), 6.61-6.50 (m , 1H), 5.99 (d, J=5.7Hz, 2H), 5.41-5.26 (m, 1H), 4.82-4.63 (m, 2H), 4.53-4.32 (m, 2H), 3.80-3.60 (m, 4H), 3.57-3.41 (m , 1H), 3.27-3.11 (m, 4H), 2.98 (dd, J=14.1, 6.6Hz, 3H), 2.82-2.74 (m, 1H), 2.68 (s, 1H), 1.32 (t, J=7.1Hz, 3H), 1.26 (dd , J=18.2, 7.3Hz, 3H), 1.17 (dd, J=14.8, 7.1Hz, 3H), 0.96 (d, J=6.6Hz, 2H), 0.81 (t, J=7.3Hz, 3H), 0.71 (t, J=7.3Hz, 3H).

[0165] Example 11

[0166] 1-(((((S)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0167]

[0168] Step A: ((S)-1,4-dioxan-2-yl)methyl (1-chloroethyl) carbonate

[0169]

[0170] Under ice bath, (S)-(1,4-dioxane-2-yl)methanol (280 mg, 2.38 mmol) and pyridine (225 mg, 2.85 mmol) were added to 10 ml of dry dichloromethane, and 1-chloroethyl chloroformate (408 mg, 2.85 mmol) was added dropwise under ice bath. After the addition was completed, the temperature was raised to room temperature and the reaction was carried out for 2 hours.

[0171] After the reaction, the mixture was poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to give 219 mg of colorless oily product ((S)-1,4-dioxan-2-yl)methyl (1-chloroethyl) carbonate (yield: 41.0%).

[0172] Step B: Synthesis of 1-(((((S)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

[0173]

[0174] At room temperature, atrasentan (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylic acid (100 mg, 0.20 mmol), ((S)-1,4-dioxan-2-yl)methyl(1-chloroethyl)carbonate (44.8 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol) and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF and heated to 65°C for 4 hours.

[0175] After the reaction, the mixture was cooled to room temperature and poured into 40 ml of ice-water solution, extracted with dichloromethane (20 ml × 3). The organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, and concentrated to dryness under reduced pressure. The crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to give 92 mg of colorless oily product 1-(((((S)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate (yield: 65.8%).

[0176] LC-MS: RT = 2.14 min, [M+H] + =699.43. 1H NMR (400MHz, DMSO) δ7.24 (dd, J=11.6, 8.7Hz, 2H), 7.06 (d, J=10.7Hz, 1H), 6.91 (dd, J=8.6, 2.7Hz, 2H), 6.85-6.70 ( m, 2H), 6.59-6.53 (m, 1H), 5.99 (d, J=6.2Hz, 2H), 4.09-3.97 (m, 2H), 3.75-3.65 (m, 6H), 3.56 (dd, J=22.9, 9.5Hz, 3H ), 3.47-3.38 (m, 2H), 3.23 (dd, J=31.0, 20.1Hz, 5H), 2.96 (dt, J=16.3, 8.0Hz, 3H), 2.84-2.62 (m, 2H), 1.33 (t, J=8. 2Hz, 3H), 1.29-1.20 (m, 3H), 1.19-1.07 (m, 3H), 0.96 (d, J=7.4Hz, 2H), 0.81 (t, J=7.3Hz, 3H), 0.71 (t, J=7.3Hz, 3H).

[0177] Example 12

[0178] Microsomal studies of compounds

[0179] (1) Experimental materials

[0180] Human liver microsomes were purchased from Red Liver Disease Research (Shanghai) Co., Ltd.

[0181] Reagents: DMSO (dimethyl sulfoxide), acetonitrile, formic acid, and propranolol (internal standard) are all commercially available.

[0182] Instrument: Thermo Fisher LC-MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer).

[0183] (2) Experimental methods

[0184] Accurately weigh a certain amount of compound and dissolve it in DMSO to prepare a 10 mM stock solution. Dilute the stock solution to a 100 μM working solution with diluent (ACN:H2O = 1:1). Then dilute it to a 3 μM dosing solution with 0.1 M potassium phosphate buffer for later use. Add 75 μL of liver microsomes to 925 μL of 0.1 M potassium phosphate buffer, mix well, and prepare a 1.5 mg / mL liver microsomal suspension. Preincubate at 37°C for 10 min. For point 0 preparation: Add 15 μL of the above liver microsomal suspension to 6 mM NADPH solution, immediately add 150 μL of propranolol acetonitrile solution for precipitation, then add 15 μL of the above dosing solution, mix well, and set aside. For 20-min and 60-min sample preparation: Add 15 μL of the dosing solution to 15 μL of the liver microsomal suspension and 15 μL of the 6 mM NADPH solution, mix well, and incubate at 37°C for 20 and 60 min, respectively. All samples were prepared in duplicate. At the relevant time points, 150 μL of propranolol acetonitrile solution was added to terminate the reaction. All samples were centrifuged at 4000 rpm for 5 minutes. 100 μL of the supernatant was added to 100 μL of ultrapure water and mixed before LC-MS / MS analysis. LC-MS / MS analysis conditions were as follows:

[0185] Chromatographic column: Waters ACQUITY™ PREMIER HSS T3, 50*2.1 mm, 1.8 μm.

[0186] Mobile phase: Water (0.1% formic acid)-acetonitrile Gradient elution according to the table below

[0187]

[0188]

[0189] (3) Data processing

[0190] Taking the initial 0 point as 100%, the relative remaining content of the drug at each time point was calculated, and taking the atrasentan at each time point as 100%, the relative amount of the example compound converted to atrasentan was calculated. The results are shown in Tables 1 and 2.

[0191] Table 1. Changes of prototypes of various drugs in microsomes

[0192]

[0193]

[0194] Table 2. Amount of Example Compounds Converted into Atrasentan in Microsomes

[0195]

[0196] The results showed that the compound of Example 3 could be rapidly metabolized in human microsomes and could be completely converted into atrasentan.

[0197] Example 13

[0198] Pharmacokinetic study of the compound in rats

[0199] (1) Experimental materials

[0200] SD rats: male, 200-300 g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0201] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), normal saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) are all commercially available.

[0202] Instrument: Thermo Fisher LC-MS / MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer).

[0203] (2) Experimental methods

[0204] The compound was weighed and dissolved in DMSO-PEG-400-saline (5:60:35, v / v / v). After oral administration to rats, 200 μL of venous blood was collected at 15 minutes, 30 minutes, 1 hour, 2 hours, 5 hours, 7 hours, and 24 hours after administration into heparinized EP tubes with sodium fluoride. The blood was centrifuged at 12,000 rpm for 2 minutes, and plasma was frozen at -80°C for analysis. A precisely weighed amount of the test compound was dissolved in DMSO to 2 mg / mL to prepare a stock solution. An appropriate amount of the stock solution was accurately aspirated and diluted with acetonitrile to prepare a series of standard solutions. 20 μL of each standard solution was accurately aspirated and added to 180 μL of blank plasma. The mixture was vortexed to mix thoroughly. Plasma samples were prepared at plasma concentrations equivalent to 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL. Duplicate samples were analyzed at each concentration to construct a standard curve. Take 30 μL of plasma, add 200 μL of acetonitrile solution of internal standard propranolol (50 ng / mL), vortex mix, add 100 μL of purified water, vortex mix again, centrifuge at 4000 rpm for 5 minutes, and collect the supernatant for LC-MS / MS analysis. LC-MS / MS detection conditions are as follows:

[0205] Chromatographic column: Waters ACQUITY™ PREMIER HSS T3, 50*2.1 mm, 1.8 μm.

[0206] Mobile phase: Water (0.1% formic acid)-acetonitrile Gradient elution according to the table below

[0207] Time (min) Water (containing 0.1% formic acid) Acetonitrile 0 85％ 15％ 0.6 85％ 15％ 1 20％ 80％ 2.3 20％ 80％ 2.31 85％ 15％ 3 85％ 15％

[0208] (3) Data processing

[0209] After LC-MS / MS detection of blood drug concentration, WinNonlin 6.1 software was used to calculate the pharmacokinetic parameters using the non-compartmental model method. The results are shown in Table 3. Table 3 Pharmacokinetic parameters of atrasentan after oral administration of atrasentan, Example 3, and Example 4 to SD rats

[0210]

[0211] Note: The dosage is converted to atrasentan

[0212] The exposure levels of the compounds of Examples 3 and 4 in rats were higher than that of atrasentan at the same dose, and their absorption after oral administration was better than that of atrasentan.

[0213] Example 14

[0214] Pharmacokinetic study of the compound in rats

[0215] (1) Experimental materials

[0216] SD rats: male, 200-300 g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0217] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), normal saline, heparin, acetonitrile, formic acid, and propranolol (internal standard) are all commercially available.

[0218] Instrument: Thermo Fisher LC-MS / MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer).

[0219] (2) Experimental methods

[0220] The compound was weighed and dissolved in DMSO-PEG-400-saline (5:60:35, v / v / v). After oral administration to rats, 200 μL of venous blood was collected at 15 minutes, 30 minutes, 1 hour, 2 hours, 5 hours, 7 hours, and 24 hours after administration into heparinized EP tubes with sodium fluoride. The blood was centrifuged at 12,000 rpm for 2 minutes, and plasma was frozen at -80°C for analysis. A precisely weighed amount of the test compound was dissolved in DMSO to 2 mg / mL to prepare a stock solution. An appropriate amount of the stock solution was accurately aspirated and diluted with acetonitrile to prepare a series of standard solutions. 20 μL of each standard solution was accurately aspirated and added to 180 μL of blank plasma. The mixture was vortexed to mix thoroughly. Plasma samples were prepared at plasma concentrations equivalent to 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL. Duplicate samples were analyzed at each concentration to construct a standard curve. Take 30 μL of plasma, add 200 μL of acetonitrile solution of internal standard propranolol (50 ng / mL), vortex mix, add 100 μL of purified water, vortex mix again, centrifuge at 4000 rpm for 5 minutes, and collect the supernatant for LC-MS / MS analysis. LC-MS / MS detection conditions are as follows:

[0221] Chromatographic column: Waters ACQUITY™ PREMIER HSS T3, 50*2.1 mm, 1.8 μm.

[0222] Mobile phase: Water (0.1% formic acid)-acetonitrile Gradient elution according to the table below

[0223] Time (min) Water (containing 0.1% formic acid) Acetonitrile 0 85％ 15％ 0.6 85％ 15％ 1 20％ 80％ 2.3 20％ 80％ 2.31 85％ 15％ 3 85％ 15％

[0224] (3) Data processing

[0225] After LC-MS / MS detection of blood drug concentration, WinNonlin 6.1 software was used to calculate the pharmacokinetic parameters using the non-compartmental model method. The results are shown in Table 4. Table 4 Pharmacokinetic parameters of atrasentan after oral administration of atrasentan to SD rats in Example 7

[0226]

[0227] The exposure of the compound of Example 7 in rats was significantly lower than that of atrasentan at the same dosage, and its absorption after oral administration was lower than that of atrasentan.

[0228] Based on Examples 12, 13 and 14, in general, the compound of Example 3 can be rapidly converted into atrasentan both in vivo and in vitro, and the exposure is higher than that of atrasentan under oral equimolar doses.

[0229] Example 15

[0230] Preparation Example of Oxalate Form A

[0231] The free base of compound A (3.5 g) was dissolved in isopropanol (10 ml), and oxalic acid dihydrate (689 mg) was added. The reaction was stirred at room temperature for 1 h and concentrated to obtain an oil. Isopropyl ether (50 ml) was added under ice bath and stirred to precipitate a solid. The mixture was stirred under ice bath for 1 day and filtered to obtain an oxalate solid (Form A).

[0232] The X-ray diffraction pattern of the obtained oxalate crystal form A is as follows: Figure 1 As shown, the specific characteristic absorption peaks are: 4.96°, 5.32°, 6.44°, 7.44°, 13.33°, 19.41°, 19.81°, 20.22°, 20.89°, and 22.48°, with an error of ±0.2°.

[0233] DSC spectrum Figure 3 As shown, the differential thermal analysis spectrum has an endothermic peak at 76.7°C with an error of ±5°C.

[0234] Example 16

[0235] Preparation Example of Oxalate Form A

[0236] Add atrasentan hydrochloride (83 g), cesium carbonate (98.8 g), potassium iodide (25.28 g), and DMF (415 mL) to a reaction flask and heat to 55°C with stirring. Add 1-chloromethyl isopropyl carbonate dropwise, and after addition, keep warm for 2 h. Once the reaction is complete, cool to below 30°C, add water (400 mL) and ethyl acetate (400 mL), stir to dissolve, allow to stand, and extract once with ethyl acetate (200 mL). Combine the organic phases, wash with saturated brine (400 mL x 3), dry over anhydrous sodium sulfate, filter, and concentrate to dryness to obtain a light yellow oily product, which is used directly in the next salt formation reaction.

[0237] To a three-necked flask, add isopropyl ether (996 g), ethyl acetate (83 g), and 13.65 g of oxalic acid in sequence, and stir to dissolve to obtain oxalic acid solution A, which is then cooled to -5°C and set aside.

[0238] Add the oil and isopropyl ether (249 g) to a single-necked flask and stir to dissolve. Slowly add the solution dropwise to the oxalic acid solution A. After the addition is complete, stir at room temperature for at least 48 hours. Filter to obtain a white solid product (Form A).

[0239] The X-ray diffraction pattern of the obtained oxalate crystal form A is as follows: Figure 2 As shown, the specific characteristic absorption peaks are: 4.96°, 5.29°, 6.45°, 7.44°, 13.35°, 19.47°, 19.89°, 20.20°, 20.90°, and 22.54, with an error of ±0.2°.

[0240] in, Figure 1 and Figure 2 The comparison of the main absorption peaks is shown in Table 1 below, with an error of ±0.2°.

[0241]

[0242]

[0243] Wherein, No. = serial number, Rel.Int. = Relative Intensity, Pos.[°2Th.] = Position[°2Theta], and the error is ±0.2°. Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form.

[0244] Summary: According to Figure 1 and Figure 2 The XRD spectrum and characteristic peak data are expressed in 2θ angles, with the strongest characteristic absorption peak at 4.96°, an error of ±0.2°, and a relative absorption intensity of 100%.

[0245] Furthermore, the crystal form has characteristic peaks at 4.96°, 5.32°, 6.44°, 7.44°, and 13.33°, with an error of ±0.2° and a relative absorption intensity greater than 15%;

[0246] Further, the crystal form has characteristic peaks at 4.96°, 5.32°, 6.44°, 7.44°, 13.33°, 19.41°, 19.81°, 20.22°, 20.89°, and 22.48°, with an error of ±0.2° and a relative absorption intensity greater than 10%, which can distinguish other substances from representing this crystal form in more detail. Other weak absorption peaks may vary significantly due to experimental operation errors. For those skilled in the art, other absorption peaks are considered unnecessary absorption peaks when characterizing this crystal form.

[0247] Example 17

[0248] Preparation Example of Oxalate Crystal Form B

[0249] The free base of compound A (2.4 g) and oxalic acid dihydrate (46 mg) were salified in ethyl acetate (1 w / w), and the mixture was added dropwise to isopropyl ether (15 w / w) at -5°C. The mixture was stirred for 1 day and filtered and dried to obtain a solid (Form B).

[0250] The X-ray diffraction pattern of the obtained oxalate crystal form B is as follows: Figure 4As shown, the specific characteristic absorption peaks are: 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 10.40°, 12.76°, 13.25°, 14.56°, 15.75°, 16.19°, 18.05°, 19.02°, 20.85°, 21.65°, 22.38°, 22.76°, and 25.21°, with an error of ±0.2°.

[0251] DSC spectrum Figure 6 As shown, the differential thermal analysis spectrum has an endothermic peak at 69.4°C with an error of ±5°C.

[0252] Example 18

[0253] Preparation Example of Oxalate Crystal Form B

[0254] Compound A oxalate Form A was dissolved in ethyl acetate (4w / w) and added dropwise to n-hexane (60w / w) at -5°C to precipitate a solid, which was transferred to a refrigerator and frozen overnight. The solid was filtered and dried under vacuum to obtain a solid (Form B).

[0255] The X-ray diffraction pattern of the obtained oxalate crystal form B is as follows: Figure 5 As shown, the specific characteristic absorption peaks are: 3.68°, 5.02°, 6.13°, 7.14°, 8.46°, 10.34°, 12.69°, 13.20°, 14.51°, 15.68°, 16.04°, 18.04°, 18.94°, 20.82°, 21.50°, 22.28°, 22.59°, and 25.11, with an error of ±0.2°.

[0256] in, Figure 4 and Figure 5 The comparison of the main absorption peaks is shown in Table 1 below, with an error of ±0.2°.

[0257]

[0258]

[0259] Wherein, No. = serial number, Rel.Int. = Relative Intensity, Pos.[°2Th.] = Position[°2Theta], and the error is ±0.2°. Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form.

[0260] Summary: According to Figure 4 and Figure 5The XRD spectrum and characteristic peak data are expressed in 2θ angles, with the strongest characteristic absorption peak at 5.01°, an error of ±0.2°, and a relative absorption intensity of 100%.

[0261] Further, the crystal form has characteristic peaks at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 13.25°, 14.56°, 18.05°, 19.02°, and 21.65°, with an error of ±0.2° and a relative absorption intensity greater than 15%;

[0262] Further, the crystal form has characteristic peaks at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 10.40°, 12.76°, 13.25°, 14.56°, 15.75°, 16.19°, 18.05°, 19.02°, 20.85°, 21.65°, 22.38°, 22.76°, and 25.21°, with an error of ±0.2° and a relative absorption intensity greater than 10%, which can distinguish other substances representing the present crystal form in more detail. Other weak absorption peaks may vary significantly due to experimental operation errors. For those skilled in the art, other absorption peaks are all absorption peaks that can be considered unnecessary when characterizing the present crystal form.

[0263] Example 19

[0264] Preparation Example of Oxalate Crystal Form C

[0265] The free base of compound A (2.2 g) was reacted with oxalic acid (70 mg) in isopropanol (2 ml). After the reaction was completed, the mixture was concentrated and isopropyl acetate (0.5 ml) was added. Isopropyl ether (15 ml) was added at -15°C and stirred for 4 h. The mixture was filtered to obtain a solid (Form C).

[0266] The X-ray diffraction pattern of the obtained oxalate crystal form C is as follows: Figure 7 As shown, the specific characteristic absorption peaks are at 5.11°, 6.94°, 8.84°, and 13.16°, with an error of ±0.2°.

[0267] DSC spectrum Figure 9 As shown, the differential thermal analysis spectrum has an endothermic peak at 63.3°C with an error of ±5°C.

[0268] Example 20

[0269] Preparation Example of Oxalate Crystal Form C

[0270] At room temperature, compound A oxalate Form A (3 g) was dissolved in acetone (2 ml) and the temperature was lowered to -5°C. The reaction solution was added dropwise to isopropyl ether (30 ml) to precipitate a solid. The mixture was stirred at low temperature for 5 h, filtered, and dried in vacuo to obtain a solid (Form C).

[0271] The X-ray diffraction pattern of the obtained oxalate crystal form C is as follows: Figure 8 As shown, the specific characteristic absorption peaks are at 5.12°, 6.92°, 8.99°, and 13.07°, with an error of ±0.2°.

[0272] in, Figure 7 and Figure 8 The comparison of the main absorption peaks is shown in Table 1 below, with an error of ±0.2°.

[0273]

[0274]

[0275] Wherein, No. = serial number, Rel.Int. = Relative Intensity, Pos.[°2Th.] = Position[°2Theta], and the error is ±0.2°. Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form.

[0276] Summary: According to Figure 7 and Figure 8 The XRD spectrum and characteristic peak data are expressed in 2θ angles, with the strongest characteristic absorption peak at 5.11°, an error of ±0.2°, and a relative absorption intensity of 100%.

[0277] Furthermore, the crystal form has characteristic peaks at 5.11°, 6.94°, 8.84°, and 13.16°, with an error of ±0.2° and a relative absorption intensity greater than 5%, which can more clearly distinguish other substances as representing this crystal form. Other weak absorption peaks may vary significantly due to experimental operation errors. For those skilled in the art, these other absorption peaks are considered unnecessary absorption peaks when characterizing this crystal form.

[0278] Example 21

[0279] Preparation Example of Phosphate Crystal Form A

[0280] Compound A free base (2.33 g), ethyl acetate (4.66 g) for dissolution, and anhydrous phosphoric acid (0.356 g) were added to a single-necked bottle in sequence, and stirred at room temperature for 30 min to prepare a phosphate solution of Compound A.

[0281] In another single-necked bottle, add isopropyl ether (23.3 g), cool to -5-5°C, and dropwise add the phosphate solution of Compound A. After the addition is complete, stir at this temperature for at least 5 h, and filter to obtain a white solid product (1.43 g).

[0282] The X-ray diffraction pattern of the obtained phosphate crystal form A is as follows: Figure 10 As shown, the specific characteristic absorption peaks are at 5.07°, 7.05°, 8.63°, and 13.25°, with an error of ±0.2°.

[0283] Example 22

[0284] Preparation Example of Phosphate Crystal Form A

[0285] The free base of compound A (4.1 g) was dissolved in isopropanol (12 ml), and phosphoric acid solution (800 mg) was added to react for 1 h. The mixture was concentrated to obtain an oily substance. Isopropyl ether (80 ml) was added under ice bath and stirred for 4 h to obtain a phosphate solid.

[0286] The X-ray diffraction pattern of the obtained phosphate crystal form A is as follows: Figure 11 As shown, the specific characteristic absorption peaks are at 5.04°, 7.04°, 8.50°, and 13.06°, with an error of ±0.2°.

[0287] in, Figure 10 and Figure 11 The comparison of the main absorption peaks is shown in Table 1 below, with an error of ±0.2°.

[0288]

[0289] Wherein, No. = serial number, Rel.Int. = Relative Intensity, Pos.[°2Th.] = Position[°2Theta], and the error is ±0.2°. Rel.Int. = Relative Intensity only indicates the approximate intensity of the characteristic peak and should not be used as a limitation on the specific crystal form.

[0290] Summary: According to Figure 10 and Figure 11 The XRD spectrum and characteristic peak data are expressed in 2θ angles, with the strongest characteristic absorption peak at 5.07°, an error of ±0.2°, and a relative absorption intensity of 100%.

[0291] Furthermore, the crystal form has characteristic peaks at 5.07°, 7.05°, 8.63°, and 13.25°, with an error of ±0.2° and a relative absorption intensity greater than 5%, which can more clearly distinguish other substances as representing this crystal form. Other weak absorption peaks may vary significantly due to experimental operation errors. For those skilled in the art, these other absorption peaks are considered unnecessary absorption peaks when characterizing this crystal form.

[0292] Example 18

[0293] Preparation of hydrochloride

[0294] Weigh Compound A free base (202.5 mg) into a 5 mL vial and add MeOH (1.0 mL) (molar ratio of methanolic hydrochloric acid). Suspend and stir at -20°C for 1 day. Rotary evaporation removes methanol. The sample becomes a gelatinous substance, which remains after vacuum drying. Add isopropyl ether (2.0 mL), suspend and stir at -20°C for 1 day. A solid precipitates, which is then centrifuged and vacuum dried to obtain a solid.

[0295] NMR spectra such as Figure 12 shown.

[0296] Example 19

[0297] Preparation of sulfate

[0298] Compound A free base (203.1 mg) was weighed into a 5 mL vial. IPA (1.0 mL) and an equimolar ratio of sulfuric acid were added. The mixture was suspended and stirred at -20°C for 1 day. The IPA was then removed by vacuum drying, resulting in a gel-like sample. Isopropyl ether (2.0 mL) was added, and the mixture was suspended and stirred at -20°C for 1 day. A solid precipitated, which was then centrifuged and dried under vacuum to obtain a solid.

[0299] NMR spectra such as Figure 13 shown.

[0300] Example 20

[0301] Stability studies

[0302] The sulfate solid, hydrochloride solid, phosphate crystal form A and oxalate crystal forms A, B and C were placed under high temperature 40°C, high humidity 92.5% RH and light (4500Lx±500Lx) conditions for 10 days, and the chemical stability of the samples was tested by HPLC.

[0303]

[0304] As can be seen from the above table, the oxalate and phosphate crystals obtained in the present invention have good stability and are superior to those of hydrochloride and sulfate.

[0305] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. A crystalline form of the compound represented by (I) and a pharmaceutically acceptable salt thereof, characterized in that: Wherein: the M is selected from oxalic acid, wherein n=1; x=1; The crystal form is oxalate crystal form A, and in the X-ray diffraction pattern, characteristic peaks are expressed in 2θ angles at 4.96°, 5.32°, 6.44°, 7.44°, 13.33°, 19.41°, 19.81°, 20.22°, 20.89°, and 22.48°, with an error of ±0.2°.

2. The compound of claim 1 or its pharmaceutically acceptable salt in a crystalline form of (I), wherein: The X-ray powder diffraction spectrum of the oxalate salt form A is shown in Figure 1 or Figure 2.

3. The compound of claim 1 or its pharmaceutically acceptable salt in a crystalline form of (I), wherein: The differential thermal analysis spectrum of the oxalate crystal form A has an endothermic peak at 76.7±5°C.

4. The compound of claim 3 in a crystalline form (I) and a pharmaceutically acceptable salt thereof, wherein: The differential thermal analysis spectrum of the oxalate crystal form A is shown in FIG3 .

5. A crystalline form of the compound represented by (I) and a pharmaceutically acceptable salt thereof, characterized in that: The crystal form is oxalate crystal form B, and in the X-ray diffraction pattern, characteristic peaks are expressed as 2θ angles at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 13.25°, 14.56°, 18.05°, 19.02°, and 21.65°, with an error of ±0.2°.

6. The compound of claim 5, wherein the crystalline form of the compound (I) and a pharmaceutically acceptable salt thereof is: The oxalate salt form B has characteristic peaks at 3.66°, 5.01°, 6.15°, 7.17°, 8.51°, 10.40°, 12.76°, 13.25°, 14.56°, 15.75°, 16.19°, 18.05°, 19.02°, 20.85°, 21.65°, 22.38°, 22.76°, and 25.21° in the X-ray diffraction pattern with a 2θ angle, with an error of ±0.2°.

7. The compound of claim 5, wherein the crystalline form of (I) and a pharmaceutically acceptable salt thereof is characterized in that: The X-ray powder diffraction spectrum of the oxalate crystal form B is shown in Figure 4 or Figure 5.

8. The compound of claim 5, wherein the crystalline form of the compound (I) and a pharmaceutically acceptable salt thereof is: The differential thermal analysis spectrum of the oxalate crystal form B has an endothermic peak at 69.4±5°C.

9. The compound of claim 5, wherein the crystalline form of (I) and a pharmaceutically acceptable salt thereof is characterized in that: The differential thermal analysis spectrum of the oxalate crystal form B is shown in FIG6 .

10. A crystalline form of the compound represented by (I) and a pharmaceutically acceptable salt thereof, characterized in that: The crystal form is oxalate crystal form C, and the X-ray powder diffraction spectrum of the oxalate crystal form C is shown in Figure 7 or Figure 8.

11. The compound of claim 10 in a crystalline form and a pharmaceutically acceptable salt thereof, characterized in that: The differential thermal analysis spectrum of the oxalate crystal form C has an endothermic peak at 63.3±5°C.

12. The compound of claim 11 in a crystalline form and a pharmaceutically acceptable salt thereof, characterized in that: The differential thermal analysis spectrum of the oxalate crystal form C is shown in FIG9 .

13. A crystalline form of the compound represented by (I) and a pharmaceutically acceptable salt thereof, characterized in that: Wherein: the M is selected from phosphoric acid, wherein n=3; x=5; The crystal form is phosphate crystal form A, and the X-ray powder diffraction spectrum of the phosphate crystal form A is shown in Figure 10 or Figure 11.

14. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a therapeutically effective amount of a compound as shown in a crystalline form of (I) according to any one of claims 1 to 13 and a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

15. Use of the compound of any one of claims 1 to 13 in a crystalline form as represented by (I) and a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing diseases related to endothelin A (ETA) receptor antagonism.

16. The use according to claim 15, characterized in that The disease is selected from the group consisting of chronic kidney disease, IgA, FSGS, Alport's disease and hypertension.

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