A benzazepine-2-one compound and its preparation method and application

Abstract

The present invention relates to the field of biomedicine technology, and more specifically to a benzazepine-2-one compound, its preparation method, and application. The benzazepine-2-one compound has high efficiency in inhibiting USP16 activity, providing a safe and efficient candidate drug molecule for the targeted treatment of prostate cancer. The benzazepine-2-one compound has a simple and easy preparation process and can be prepared in a single step via a click reaction. According to the experimental results in the examples of the present invention, the tumor inhibition rate of the tumor-bearing group of mice treated with the benzazepine-2-one compound was superior to that of the positive control group, indicating that the benzazepine-2-one compound can be used to treat prostate cancer and has good development prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a benzazepine-2-one compound, a preparation method and an application thereof. Background Art

[0002] Prostate cancer is the second most common cancer in men worldwide. Drug resistance in the castration-resistant prostate (CRPC) stage presents a therapeutic bottleneck. Research has shown that ubiquitin-specific protease 16 (USP16) specifically removes the ubiquitination modification of c-Myc, inhibiting its proteasomal degradation and thereby stabilizing c-Myc expression. As a core oncogenic factor, c-Myc activates cell cycle genes (such as Cyclin D1 and CDK4), driving cancer cell proliferation and invasion. Inhibiting USP16 significantly reduces c-Myc levels and inhibits tumor growth in CRPC cell lines and xenograft models. Clinical analyses have shown that USP16 is highly expressed in prostate cancer tissues and positively correlates with tumor malignancy (Gleason score), suggesting its potential as a prognostic marker and therapeutic target.

[0003] While existing broad-spectrum USP inhibitors (such as YM155) can inhibit USP16 activity, their lack of selectivity can lead to off-target effects (such as abnormal DNA damage and immune imbalance), limiting their clinical application. Developing highly selective USP16 inhibitors is crucial. Such drugs can be used alone to treat CRPC or in combination with second-generation AR antagonists (such as enzalutamide) to enhance anticancer efficacy by synergistically blocking AR signaling and the c-Myc pathway. Furthermore, USP16 inhibitors also have potential for treating solid tumors such as liver cancer and colon cancer, where USP16 is abnormally activated.

[0004] Therefore, it is necessary to develop new USP16 selective inhibitors. Summary of the Invention

[0005] In order to promote the therapeutic effect of prostate cancer, the present invention provides a benzazepine-2-one compound and its preparation method and application.

[0006] The first aspect of the present invention provides a benzazepin-2-one compound or a pharmaceutically acceptable salt thereof. The benzazepin-2-one compound has a structure shown in Formula 3:

[0007] ;

[0008] Wherein, R is selected from a hydrogen atom, a C1-C4 alkoxy group, a halogen atom, and a trifluoromethyl group; and X is a carbon atom or a nitrogen atom.

[0009] The above-mentioned benzazepin-2-one compound or a pharmaceutically acceptable salt thereof can selectively inhibit USP16 and is a USP16 selective inhibitor. The pharmaceutically acceptable salt of the above-mentioned benzazepin-2-one compound can be easily prepared from the above-mentioned benzazepin-2-one compound as a raw material. For example, the hydrochloride salt of the above-mentioned benzazepin-2-one compound can be prepared by reacting the above-mentioned benzazepin-2-one compound with hydrochloric acid.

[0010] In some optional embodiments, the above R is selected from hydrogen, methoxy, ethoxy, fluorine atom, chlorine atom, bromine atom, iodine atom, and trifluoromethyl.

[0011] In some optional embodiments, the above-mentioned benzazepine-2-one compound has a structure shown in one of the following formulae:

[0012] 、 、 、 、 、 、 .

[0013] The second aspect of the present invention provides a method for preparing the above-mentioned benzazepine-2-one compound, which comprises the following steps:

[0014] reacting the compound represented by Formula 1 with the compound represented by Formula 2 to obtain the above-mentioned benzazepine-2-one compound;

[0015] 、 ;

[0016] Wherein, R is selected from a hydrogen atom, a C1-C4 alkoxy group, a halogen atom, and a trifluoromethyl group; and X is a carbon atom or a nitrogen atom.

[0017] In some optional embodiments, the above reaction is carried out with the participation of copper sulfate and vitamin C.

[0018] In some optional embodiments, the above reaction is carried out at 20-50° C. Preferably, the above reaction is carried out at room temperature.

[0019] In some optional embodiments, the above reaction is carried out in an organic solvent. Preferably, the above organic solvent is a mixture of isopropyl alcohol and water.

[0020] The third aspect of the present invention provides the use of the above-mentioned benzazepin-2-one compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating and / or preventing prostate cancer.

[0021] A fourth aspect of the present invention provides a drug for treating and / or preventing prostate cancer, comprising the aforementioned benzazepin-2-one compound or a pharmaceutically acceptable salt thereof. The aforementioned benzazepin-2-one compound or a pharmaceutically acceptable salt thereof is the main active ingredient (primary agent) in the drug for treating and / or preventing prostate cancer.

[0022] In some optional embodiments, the drug for treating and / or preventing prostate cancer further comprises an excipient. The drug for treating and / or preventing prostate cancer can be in any pharmaceutically acceptable dosage form. The excipient is stable, has no incompatibility with the main drug, does not produce side effects, does not affect efficacy, is not easily deformed, cracked, or moldy at room temperature, and is harmless to the human body.

[0023] In some optional embodiments, the excipient is at least one of gum arabic, syrup, lanolin, and starch.

[0024] The technical solution of the embodiment of the present invention has the following beneficial effects:

[0025] The present invention obtains a novel USP16 selective inhibitor, namely the above-mentioned benzazepin-2-one compound, which has the ability to effectively inhibit USP16 activity and provides a safe and efficient candidate drug molecule for the targeted treatment of prostate cancer; the preparation process of the benzazepin-2-one compound is simple and easy, and can be prepared in one step by a click reaction; according to the experimental results in the examples of the present invention, the tumor inhibition rate of the tumor-bearing group of mice treated with the benzazepin-2-one compound is better than that of the positive control group, indicating that the benzazepin-2-one compound can be used to treat prostate cancer and has good development prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a photograph of mouse tumor tissue 14 days after the experiment on the inhibition of prostate tumor by the benzazepine-2-one compound in Example 14 of the present invention.

[0027] Figure 2 This is a graph showing the changes in tumor volume in mice undergoing an experimental study on the inhibition of prostate tumors by the benzazepine-2-one compound in Example 14 of the present invention. DETAILED DESCRIPTION

[0028] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0029] In the following examples, the synthesis method of the azide compound represented by Formula 2 is as follows:

[0030]

[0031] Under an argon atmosphere, the compound of Formula 4 (1.0 mmol, 1.0 eq.) and ammonium acetate (3.0 mmol, 3.0 eq.) were dissolved in 10.0 mL of methanol. The temperature was cooled to 0°C, and sodium cyanoborohydride (10.0 mmol, 10.0 eq.) was added in small portions. The system was then transferred to an oil bath and heated to reflux. After completion of the reaction, saturated sodium carbonate solution was added to quench the reaction. The product was extracted with ethyl acetate, and the organic phases were combined, dried, concentrated, and purified by column chromatography to obtain the compound of Formula 5 in a 31% yield.

[0032] The characterization results of the compound shown in Formula 5 are as follows: 1 H NMR (400 MHz, CDCl3) δ 8.70 (s, 1H),7.72 (ddd, J = 7.5, 2.0, 1.0 Hz, 1H), 7.24 (td, J = 7.4, 2.0 Hz, 1H), 7.11(td, J = 7.5, 2.1 Hz, 1H), 7.06 (dd, J = 7.3, 2.0 Hz, 1H), 4.13 (td, J = 7.0,1.1 Hz, 1H), 2.72 – 2.56 (m, 1H), 2.45 – 2.18 (m, 2H), 2.20 – 1.97 (m, 1H),1.36 (s, 2H). 13 C NMR (100 MHz, CDCl3) δ 174.01, 136.69, 128.27, 127.66, 125.90, 122.37, 121.72, 52.43, 31.86, 30.29.

[0033] The compound represented by Formula 5 (1.0 mmol, 1.0 eq.) and the compound represented by Formula 6 (1.0 mmol, 1.0 eq.) obtained above were dissolved in 10.0 mL of a mixture of methyl tert-butyl ether and N,N-dimethylformamide (volume ratio of 1:1). Potassium bicarbonate (3.0 mmol, 3.0 eq.) was then added to the mixture and allowed to react at room temperature. After completion of the reaction, ethyl acetate was added for extraction. The organic phase was dried and concentrated to obtain the azide compound represented by Formula 2.

[0034] The characterization results of the azide compound shown in Formula 2 are as follows: 1H NMR (400 MHz, CDCl3) δ 8.70 (s,1H), 7.41 (ddd, J = 7.5, 2.0, 0.9 Hz, 1H), 7.22 (td, J = 7.5, 2.0 Hz, 1H),7.14 (td, J = 7.4, 2.1 Hz, 1H), 7.04 (dd, J = 7.4, 2.1 Hz, 1H), 3.67 (td, J =7.0, 1.1 Hz, 1H), 2.66 (dt, J = 13.9, 7.1 Hz, 1H), 2.28 (dt, J = 13.7, 7.0Hz, 1H), 2.23 – 2.10 (m, 1H), 2.06 – 1.94 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ174.01, 136.37, 134.14, 128.03, 127.13, 125.80, 123.23, 55.90, 31.60, 28.49.

[0035] Among them, the racemic azide compound represented by Formula 2 is represented by Formula 2a. After chiral resolution, the R-configured azide compound represented by Formula 2 is represented by Formula 2b, and the S-configured azide compound represented by Formula 2 is represented by Formula 2c.

[0036] Example 1: Synthesis of the Benzazepine-2-one Compound of Formula 3a

[0037] Under argon, phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) and the racemic azide of Formula 2a (202.0 mg, 1.0 mmol, 1.0 eq.) were dissolved in 10 mL of a 1:1 volume ratio mixture of isopropanol and water. Copper sulfate pentahydrate (25.0 mg, 0.1 mmol, 0.1 eq.) and vitamin C (17.6 mg, 0.5 mmol, 0.5 eq.) were added and reacted at room temperature for 1 h. The reaction was quenched by the addition of saturated sodium bicarbonate, and the mixture was extracted with ethyl acetate. The organic phase was dried, concentrated, and purified by column chromatography to afford the benzazepine-2-one compound of Formula 3a (270.6 mg, 89% yield).

[0038]

[0039] The characterization results of the benzazepine-2-one compound represented by formula 3a are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.27 (s, 1H), 7.85 – 7.78 (m, 3H), 7.54 – 7.46 (m, 2H), 7.46 –7.39 (m, 1H), 7.29 – 7.22 (m, 2H), 7.17 (td, J = 7.5, 1.5 Hz, 1H), 5.94 –5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 148.45, 138.76, 131.00, 130.41, 130.19,129.41, 128.70, 128.32, 125.43, 124.57, 120.57, 119.59, 65.68, 35.37, 29.48.

[0040] Example 2: Synthesis of the Benzazepine-2-one Compound of Formula 3b

[0041] This example is basically the same as Example 1, except that the racemic azide compound represented by Formula 2a is replaced by the azide compound represented by Formula 2b with R configuration (obtained by chiral resolution of the racemic azide compound represented by Formula 2a), to obtain the benzazepine-2-one compound represented by Formula 3b (276.6 mg, yield 91%).

[0042]

[0043] The characterization results of the benzazepine-2-one compound represented by formula 3b are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.27 (s, 1H), 7.85 – 7.78 (m, 3H), 7.54 – 7.46 (m, 2H), 7.46 –7.39 (m, 1H), 7.29 – 7.22 (m, 2H), 7.17 (td, J= 7.5, 1.5 Hz, 1H), 5.94 –5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 148.45, 138.76, 131.00, 130.41, 130.19,129.41, 128.70, 128.32, 125.43, 124.57, 120.57, 119.59, 65.68, 35.37, 29.48.

[0044] Example 3: Synthesis of the Benzazepine-2-one Compound of Formula 3c

[0045] This example is basically the same as Example 1, except that the racemic azide compound represented by Formula 2a is replaced by the azide compound represented by Formula 2c with S configuration (obtained by chiral resolution of the racemic azide compound represented by Formula 2a), to obtain the benzazepine-2-one compound represented by Formula 3c (276.6 mg, yield 91%).

[0046]

[0047] The characterization results of the benzazepine-2-one compound represented by formula 3c are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.27 (s, 1H), 7.85 – 7.78 (m, 3H), 7.54 – 7.46 (m, 2H), 7.46 –7.39 (m, 1H), 7.29 – 7.22 (m, 2H), 7.17 (td, J = 7.5, 1.5 Hz, 1H), 5.94 –5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 148.45, 138.76, 131.00, 130.41, 130.19,129.41, 128.70, 128.32, 125.43, 124.57, 120.57, 119.59, 65.68, 35.37, 29.48.

[0048] Example 4: Synthesis of the Benzazepine-2-one Compound of Formula 3d

[0049] This example is basically the same as Example 2, except that phenylacetylene is replaced with 4-(trifluoromethyl)phenylacetylene to obtain the benzazepine-2-one compound represented by Formula 3d (319.9 mg, yield 86%).

[0050]

[0051] The characterization results of the benzazepine-2-one compound represented by formula 3d are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.33 (d, J = 0.7 Hz, 1H), 7.95 – 7.88 (m, 2H), 7.83 (dd, J =8.1, 1.5 Hz, 1H), 7.76 (dq, J = 11.3, 1.3 Hz, 2H), 7.29 – 7.22 (m, 2H), 7.17(td, J = 7.5, 1.5 Hz, 1H), 5.94 – 5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 –2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 147.96,138.76, 132.70, 132.44, 132.18, 131.93, 130.19, 128.86, 128.70, 128.32,127.16, 127.13, 127.09, 127.05, 125.43, 124.98, 124.42, 124.39, 124.35,124.32, 122.81, 120.63, 120.57, 119.53, 65.68, 35.37, 29.48.

[0052] Example 5: Synthesis of the Benzazepine-2-one Compound of Formula 3e

[0053] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with 4-ethynylanisole (132.2 mg, 1.0 mmol, 1.0 eq.) to obtain the benzazepine-2-one compound represented by Formula 3e (300.6 mg, 90% yield).

[0054]

[0055] The characterization results of the benzazepine-2-one compound represented by formula 3e are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.25 (d, J = 0.7 Hz, 1H), 7.83 (dd, J = 8.1, 1.5 Hz, 1H), 7.67– 7.60 (m, 2H), 7.29 – 7.22 (m, 2H), 7.17 (td, J = 7.5, 1.5 Hz, 1H), 6.91 –6.84 (m, 2H), 5.94 – 5.88 (m, 1H), 3.78 (s, 3H), 2.75 – 2.65 (m, 1H), 2.62 –2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 160.82,148.49, 138.76, 130.19, 128.70, 128.32, 125.62, 125.43, 125.38, 120.57,119.41, 115.33, 65.68, 55.35, 35.37, 29.48.

[0056] Example 6: Synthesis of the Benzazepine-2-one Compound of Formula 3f

[0057] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with p-chlorophenylacetylene (136.6 mg, 1.0 mmol, 1.0 eq.) to obtain the benzazepine-2-one compound represented by Formula 3f (311.0 mg, 92% yield).

[0058]

[0059] The characterization results of the benzazepine-2-one compound represented by formula 3f are as follows:1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.28 (d, J = 0.7 Hz, 1H), 7.83 (dd, J = 8.1, 1.4 Hz, 1H), 7.76– 7.70 (m, 2H), 7.44 – 7.38 (m, 2H), 7.29 – 7.22 (m, 2H), 7.17 (td, J = 7.5,1.5 Hz, 1H), 5.94 – 5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 148.37, 138.76, 136.12,131.05, 130.19, 130.02, 128.70, 128.32, 126.48, 125.43, 120.57, 119.55,65.68, 35.37, 29.48.

[0060] Example 7: Synthesis of the Benzazepine-2-one Compound of Formula 3g

[0061] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with p-fluorophenylacetylene (120.1 mg, 1.0 mmol, 1.0 eq.) to obtain the benzazepine-2-one compound represented by Formula 3g (302.7.0 mg, 94% yield).

[0062]

[0063] The characterization results of the benzazepine-2-one compound represented by formula 3g are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.28 (d, J = 0.7 Hz, 1H), 7.91 – 7.84 (m, 2H), 7.83 (dd, J=8.1, 1.4 Hz, 1H), 7.29 – 7.22 (m, 2H), 7.21 – 7.14 (m, 2H), 7.14 – 7.09 (m,2H), 5.94 – 5.88 (m, 1H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 –2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 165.08, 163.11, 148.72,138.76, 130.19, 128.70, 128.48, 128.45, 128.32, 126.36, 126.29, 125.43,120.57, 119.59, 116.98, 116.80, 65.68, 35.37, 29.48.

[0064] Example 8: Synthesis of the Benzazepine-2-one Compound of Formula 3h

[0065] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with 2-chloro-4-ethynyl-1-methoxybenzene (166.6 mg, 1.0 mmol, 1.0 eq.) to obtain the benzazepine-2-one compound represented by Formula 3h (320.2 mg, yield 87%).

[0066]

[0067] The characterization results of the benzazepine-2-one compound represented by formula 3h are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.23 ​​(d, J = 0.7 Hz, 1H), 7.85 – 7.79 (m, 2H), 7.64 (dd, J =8.8, 2.2 Hz, 1H), 7.29 – 7.22 (m, 2H), 7.17 (td, J = 7.5, 1.5 Hz, 1H), 7.05(d, J = 8.7 Hz, 1H), 5.94 – 5.88 (m, 1H), 3.90 (s, 3H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H).13 C NMR (100 MHz, CDCl3) δ 171.13,156.50, 148.46, 138.76, 130.19, 128.70, 128.32, 126.23, 125.64, 125.43,124.34, 122.97, 120.57, 119.78, 114.66, 65.68, 56.67, 35.37, 29.48.

[0068] Example 9: Synthesis of the Benzazepine-2-one Compound of Formula 3i

[0069] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with 4-ethynyl-1-methoxy-2-(trifluoromethyl)benzene (200.2 mg, 1.0 mmol, 1.0 eq) to obtain the benzazepine-2-one compound represented by Formula 3i (353.8 mg, 88% yield).

[0070]

[0071] The characterization results of the benzazepine-2-one compound represented by formula 3i are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.25 – 8.17 (m, 2H), 7.83 (dd, J = 8.2, 1.5 Hz, 1H), 7.61 (dd, J = 8.1, 2.2 Hz, 1H), 7.29 – 7.22 (m, 3H), 7.17 (td, J = 7.5, 1.5 Hz, 1H),5.94 – 5.88 (m, 1H), 3.85 (s, 3H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H),2.35 – 2.24 (m, 1H). 13C NMR (100 MHz, CDCl3) δ 171.13, 157.27, 157.25, 157.24,157.23, 148.41, 138.76, 130.19, 128.70, 128.32, 127.25, 125.43, 125.11,125.06, 124.62, 124.58, 124.55, 124.52, 124.24, 124.21, 124.18, 124.15,122.87, 120.67, 120.57, 119.55, 116.41, 116.15, 115.88, 115.62, 114.22,114.18, 114.15, 114.12, 65.68, 55.23, 35.37, 29.48.

[0072] Example 10: Synthesis of the Benzazepine-2-one Compound of Formula 3j

[0073] This example is basically the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with 4-ethynyl-2-fluoro-1-methoxybenzene (150.0 mg, 1.0 mmol, 1.0 eq) to obtain the benzazepine-2-one compound represented by Formula 3j (320.3 mg, 91% yield).

[0074]

[0075] The characterization results of the benzazepine-2-one compound represented by formula 3j are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.23 ​​(d, J = 0.7 Hz, 1H), 7.83 (dd, J = 8.1, 1.5 Hz, 1H), 7.56(dd, J = 9.9, 2.2 Hz, 1H), 7.49 (dd, J = 12.1, 2.1 Hz, 1H), 7.29 – 7.22 (m,2H), 7.17 (td, J = 7.5, 1.5 Hz, 1H), 7.04 (dd, J= 9.9, 4.7 Hz, 1H), 5.94 –5.88 (m, 1H), 3.86 (s, 3H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 –2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 154.25, 152.27, 148.61,148.50, 147.04, 147.02, 138.76, 130.19, 128.70, 128.32, 125.43, 123.30,123.24, 122.42, 122.38, 120.57, 119.70, 115.19, 115.14, 111.47, 111.29,65.68, 56.51, 56.48, 35.37, 29.48.

[0076] Example 11: Synthesis of the Benzazepine-2-one Compound of Formula 3k

[0077] This example is essentially the same as Example 2, except that phenylacetylene (102.0 mg, 1.0 mmol, 1.0 eq.) is replaced with 5-ethynyl-2-methoxypyridine (133.2 mg, 1.0 mmol, 1.0 eq.) to obtain the benzazepine-2-one compound of Formula 3k (301.5 mg, 90% yield).

[0078]

[0079] The characterization results of the benzazepine-2-one compound represented by formula 3k are as follows: 1 H NMR (400 MHz, CDCl3) δ9.25 (s, 1H), 8.65 (d, J = 2.0 Hz, 1H), 8.27 (d, J = 0.7 Hz, 1H), 8.03 (dd, J = 8.6, 1.8 Hz, 1H), 7.83 (dd, J = 8.1, 1.5 Hz, 1H), 7.29 – 7.22 (m, 2H), 7.17(td, J = 7.5, 1.5 Hz, 1H), 6.89 (d, J= 8.6 Hz, 1H), 5.94 – 5.88 (m, 1H), 3.92 (s, 3H), 2.75 – 2.65 (m, 1H), 2.62 – 2.49 (m, 2H), 2.35 – 2.24 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 171.13, 165.81, 147.26, 146.40, 138.76, 134.81,130.19, 128.70, 128.32, 125.43, 121.15, 120.57, 120.40, 110.98, 65.66, 54.60,35.37, 29.48.

[0080] Example 12: Benzazepin-2-one compounds inhibit USPs kinases

[0081] Deubiquitinase (DUB) activity was monitored by a fluorometric assay using ubiquitin-rhodamine 110 (Ub-Rho110, Catalog # M3022, UBP Bio) in a buffer containing 50 mM HEPES (pH 8.0), 150 mM NaCl, 0.5 mM EDTA, 1 mM DTT, and 0.1 mg / mL bovine serum albumin (BSA). The assay procedure is briefly as follows: the benzazepine-2-one compounds represented by Formulas 3a to 3k were first diluted to 10 μM, followed by the addition of a 10 nM deubiquitinase. After brief mixing, the mixture was placed in a 200 μL reaction volume and incubated at room temperature for 1 hour. After the addition of 200 nM Ub-Rho110, the final fluorescence signal (excitation wavelength: 485 nm, emission wavelength: 535 nm) was obtained using a TECAN SPARK multifunctional microplate reader. The inhibition rate of the benzazepine-2-one compound on the enzyme activity was calculated based on the fluorescence signal. The results are shown in Table 1.

[0082] Table 1 Results of the inhibitory activity of benzazepine-2-one compounds against USPs

[0083]

[0084] In Table 1, +++ represents an inhibition rate greater than 80%, ++ represents an inhibition rate between 50% and 80%, and + represents an inhibition rate less than 50%.

[0085] It can be seen that the benzazepine-2-one compounds represented by Formula 3d, Formula 3i and Formula 3j can selectively inhibit the kinase activity of USP16 at the protein level.

[0086] Example 13: Benzazepine-2-one compounds inhibit prostate cancer cells

[0087] The in vitro antiproliferative activity of the benzazepine-2-one compounds represented by Formula 3d, Formula 3i, and Formula 3j against prostate cancer cell lines PC-3 and DU145 was evaluated using a CCK-8 assay. Using UBD1031 as a positive control, a 10-μM maximum concentration was established, followed by a three-fold serial dilution cycle, with a minimum concentration of 0 μM, for a total of 10 concentrations. Logarithmically growing PC-3 and DU145 cells were resuspended and counted, and the concentration was adjusted to 5×10 3 -1×10 4 Cells / mL were added to a 96-well plate: 100 μL of cell suspension was added to each well to a cell density of 500-1000 cells / well (three replicates were set). The plate was placed in an incubator and incubated for 24 hours before adding the prepared drug (1-1000 nM) and incubating for 72 hours. CCK-8 working solution was added and incubated for another 3 hours. The plate was immediately placed in a microplate reader for reading. Data were processed using GraphPad Prism 9 to calculate the IC 50 As shown in Table 2, the benzazepine-2-one compounds represented by Formula 3d, Formula 3i and Formula 3j all have excellent in vitro anti-proliferation activity against prostate cancer cells.

[0088] Table 2 Determination of the antiproliferative activity of benzazepine-2-one compounds on prostate cancer cells

[0089]

[0090] In Table 2, + indicates >1000 nM; ++ indicates 1000-100 nM; and +++ indicates <100 nM.

[0091] Example 14: Benzazepine-2-one compounds inhibit animal tumors

[0092] To evaluate the anti-tumor activity of the active compounds in vivo, animal experiments were conducted using two prostate cancer cell xenograft models (PC-3 and DU145) in BALB / c mice. Specifically, a subcutaneous tumor-bearing mouse model was established to evaluate the anti-tumor effects of the benzazepin-2-one compounds represented by Formula 3d, 3i, and 3j in BALB / c mice. PC-3 and DU145 cells were cultured in a 37°C, 5% CO2 incubator. All mice were housed under standard specific pathogen-free (SPF) conditions. BALB / c mice aged 6-8 weeks were subcutaneously injected with PC-3 and DU145 cells. Mice were examined daily to assess tumor progression. When tumors reached a certain size, mice were randomly divided into a control group and a treatment group, with 5 mice in each group. The treatment group received an intraperitoneal injection of 50 mpk of the benzazepin-2-one compound dissolved in 100 µL of 5% DMSO and 95% saline. Tumor diameter was measured using a vernier caliper and mouse weight was recorded daily. Tumor tissue was removed on day 14 and the tumor inhibition effect of the compound was evaluated by the change in tumor volume. The results are shown in Table 3 and Figure 1-Figure 2 As shown in , the benzazepine-2-one compounds represented by Formula 3d, Formula 3i and Formula 3j all have excellent in vivo anti-prostate tumor proliferation activity.

[0093] Table 3 Animal experimental results of benzazepine-2-one compounds inhibiting prostate tumors

[0094]

[0095] In Table 3, + represents a tumor inhibition rate of less than 50%, ++ represents a tumor inhibition rate of 50%-80%, and +++ represents a tumor inhibition rate of more than 80%.

[0096] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form or substance. It should be pointed out that ordinary technicians in this technical field can make several improvements and supplements without departing from the method of the present invention. These improvements and supplements should also be regarded as the scope of protection of the present invention. Any equivalent changes, modifications and evolutions made by technicians familiar with this profession without departing from the spirit and scope of the present invention by using the technical content disclosed above are all equivalent embodiments of the present invention; at the same time, any equivalent changes, modifications and evolutions made to the above embodiments based on the essential technology of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. A benzazepine-2-one compound or a pharmaceutically acceptable salt thereof, characterized in that: The benzazepine-2-one compound has a structure shown in Formula 3, Formula 3h, Formula 3i or Formula 3j: 、 、 、 ； Wherein, R is selected from a hydrogen atom, a C1-C4 alkoxy group, a halogen atom, and a trifluoromethyl group; and X is a carbon atom or a nitrogen atom.

2. The benzazepine-2-one compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The R is selected from hydrogen, methoxy, ethoxy, fluorine, chlorine, bromine, iodine, and trifluoromethyl.

3. The benzazepine-2-one compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The benzazepine-2-one compound has a structure shown in one of the following formulas: 、 、 、 、 、 。 4. A method for preparing a benzazepine-2-one compound, characterized in that: The following steps are involved: reacting the compound represented by Formula 1, Formula 1h, Formula 1i, or Formula 1j with the compound represented by Formula 2 to obtain a benzazepine-2-one compound represented by Formula 3, Formula 3h, Formula 3i, or Formula 3j; 、 、 、 、 、 、 、 、 ； Wherein, R is selected from a hydrogen atom, a C1-C4 alkoxy group, a halogen atom, and a trifluoromethyl group; and X is a carbon atom or a nitrogen atom.

5. The method according to claim 4, characterized in that The reaction is carried out in the presence of copper sulfate and vitamin C.

6. The method according to claim 4, characterized in that The reaction is carried out at 20-50°C.

7. The method according to claim 4, characterized in that The reaction is carried out in an organic solvent.

8. Use of the benzazepin-2-one compound or a pharmaceutically acceptable salt thereof according to claim 1, 2 or 3 in the preparation of a medicament for treating and / or preventing prostate cancer.

9. A drug for treating and / or preventing prostate cancer, characterized in that: The invention comprises the benzazepine-2-one compound or a pharmaceutically acceptable salt thereof according to claim 1, 2 or 3.

10. The drug for treating and / or preventing prostate cancer according to claim 9, characterized in that The drug for treating and / or preventing prostate cancer further comprises an excipient.

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