Benzazepine derivatives, radioactive probes and their applications

Abstract

The present invention provides benzodiazepine derivatives, radioactive probes and their applications, belonging to the field of biomedicine technology. The present invention modifies the benzodiazepine derivatives and introduces hydrophilic or lipophilic side chains. The prepared benzodiazepine derivatives can be used as precursors targeting V2R, which can achieve specific targeting of V2R and provide new molecular tools for the subsequent diagnosis and treatment of V2R-positive tumors. 68 The Ga-labeled radioactive probe of the present invention exhibits good tumor imaging effects and can achieve PET imaging of V2R-positive tumors, which helps to improve the accuracy of tumor diagnosis. It can also be used to evaluate treatment effects, has a high "target / non-target" ratio, and can better display the distribution of V2R receptors throughout the body. It has many applications in the diagnosis and treatment of cancer.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to benzoazepine derivatives, radioactive probes and applications thereof. Background Art

[0002] Renal cell carcinoma (RCC) originates from renal tubular epithelial cells and accounts for over 90% of renal cancers. Clear cell renal cell carcinoma (ccRCC) accounts for the majority (70%) of RCC cases, papillary RCC (pRCC) accounts for 10-15%, and chromophobe RCC (rhRCC) accounts for approximately 5%. Most RCCs are diagnosed when the disease is confined to the kidney. These patients have a favorable prognosis, with a 5-year survival rate exceeding 90%. However, the prognosis is poor for patients with advanced disease, such as those with tumor cells that have spread to surrounding tissues or organs and / or regional lymph nodes, with a 5-year survival rate of approximately 70%. For patients with distant metastases, the 5-year survival rate is only 13%. While the vast majority of RCCs are curable with surgery, 20% to 30% of patients will develop distant metastases, and 2% to 5% will experience local recurrence. Therefore, the diagnosis and treatment of metastatic and recurrent RCC remain key priorities in RCC.

[0003] Over the past 20 years, with the implementation of personalized medicine and precision medicine, radiopharmaceuticals and fusion imaging equipment have been vigorously developed, and nuclear medicine has played a significant role in the diagnosis and treatment of various diseases. Functional PET / CT has been shown to be very important in the diagnosis, staging, and treatment guidance of various malignancies, including lymphoma, esophageal cancer, nasopharyngeal cancer, lung cancer, gastric cancer, and intestinal cancer. However, its application in RCC remains relatively limited. Compared with traditional imaging methods, FDG PET / CT has limited supplementary value in assessing RCC metastasis and recurrence. In addition, in low-grade renal cancer, lesions often have low levels of glucose metabolism and poor FDG uptake, making it difficult to effectively detect tumors. In addition, current research on RCC PET imaging agents includes 89Zr-bevacizumab, 89Zr-glinduzumab, 18F-fluoro-L-thymidine (FLT), prostate-specific membrane antigen (PSMA), 11C-choline, 18F-sodium fluoride (NaF), etc. However, due to the large molecular weight and long blood circulation time of the above-mentioned imaging agents such as 89Zr-bevacizumab and 89Zr-glinduzumab, the optimal imaging effect is usually achieved 3-7 days after injection, and their clinical application has certain limitations. PSMA is mainly expressed in the endothelial cells of RCC's newly formed capillaries and cannot directly display tumor cell activity, resulting in it not yet achieving satisfactory results in the clinical transformation of RCC diagnosis and treatment.

[0004] Integrated nuclear medicine diagnosis and treatment is one of the most active research areas in nuclear medicine technology. By using different diagnostic and therapeutic radionuclide probes to combine imaging diagnosis with internal irradiation therapy, this approach achieves both visual diagnosis and precise treatment. This approach permeates the entire process of personalized medicine, encompassing diagnosis, grading and staging, treatment, efficacy monitoring, and prognosis assessment. Therefore, it holds broad promise for application in the field of oncology. Currently, FDA-approved integrated diagnostic and therapeutic radiopharmaceuticals are available for differentiated thyroid cancer, neuroendocrine tumors, and prostate cancer. However, no corresponding radiopharmaceuticals have entered clinical trials for RCC, a cancer with a high morbidity and mortality rate. Metastatic RCC is primarily treated with systemic therapies, such as tyrosine kinase inhibitors and immunotherapy. However, for drug-resistant metastatic RCC, there are few alternative treatment options, and their efficacy and safety are uncertain.

[0005] Therefore, developing a radioactive probe with high targeting and definite efficacy will be beneficial to promoting the personalized diagnosis and precise treatment of RCC. Summary of the Invention

[0006] The purpose of the present invention is to provide benzoazepine derivatives, radioactive probes and their applications, which can be used for in vivo tumor imaging based on V2R targets in the human body and to detect the therapeutic efficacy of V2R-targeted therapies, providing an effective means for related research.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a benzodiazepine derivative, wherein the benzodiazepine derivative is a compound having a structure shown in Formula 1: Formula 1.

[0009] The present invention also provides another benzodiazepine derivative, which is a compound having a structure shown in Formula 2: Formula 2.

[0010] The present invention also provides another benzodiazepine derivative, which is a compound having a structure shown in Formula 3: Formula 3.

[0011] The present invention also provides a radioactive probe, which is a compound having a structure shown in Formula 4: Formula 4.

[0012] The present invention also provides another radioactive probe, which is a compound having a structure shown in Formula 5: Formula 5.

[0013] The present invention also provides another radioactive probe, which is a compound having a structure shown in Formula 6: Formula 6.

[0014] The present invention also provides the use of the above-mentioned benzoazepine derivatives or radioactive probes in the preparation of arginine vasopressin receptor 2 targeting drugs.

[0015] The present invention also provides the use of the above-mentioned benzoazepine derivatives or radioactive probes in the preparation of cancer cell imaging agents, cancer auxiliary diagnostic reagents, cancer staging evaluation reagents or cancer drug efficacy evaluation products.

[0016] Preferably, the cancer is renal cell carcinoma.

[0017] Beneficial effects of the present invention:

[0018] The present invention modifies benzoazepine derivatives by introducing hydrophilic or lipophilic side chains. The resulting benzoazepine derivatives can be used as precursors for V2R targeting, enabling specific V2R targeting and providing new molecular tools for the subsequent diagnosis and treatment of V2R-positive tumors. The preparation method of the benzoazepine derivatives described in the present invention is simple, and the raw materials are readily available, promising promising clinical translation prospects.

[0019] The radioactive probe of the present invention is combined with 68 Ga labeling shows good tumor imaging effects, can realize PET imaging of V2R-positive tumors, provide clear tumor images, and help improve the accuracy of tumor diagnosis; the radioactive probe of the present invention can also be used to evaluate the treatment effect, has a high "target / non-target" ratio, and can better display the distribution of V2R receptors throughout the body.

[0020] The present invention provides a benzazepine derivative and its 68 Ga markers have many applications in the diagnosis and treatment of cancer, including reflecting the expression level of V2R in renal cell carcinoma, which can be used for the diagnosis and staging of renal cell carcinoma, and formulating treatment plans at the same time, realizing the integration of targeted V2R diagnosis and treatment. It has high clinical translation value and provides a new approach for personalized diagnosis and precision treatment of tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 To determine the labeling rate of radioactive probe 1 using radioactive high performance liquid chromatography (HPLC)

[0022] Figure 2 To determine the labeling rate of radioactive probe 2 using radioactive high performance liquid chromatography (HPLC)

[0023] Figure 3 To determine the labeling rate of radioactive probe 3 using radioactive high performance liquid chromatography (HPLC)

[0024] Figure 4 The stability of radioactive probe 1 in fetal bovine serum and saline

[0025] Figure 5 The stability of radioactive probe 2 in fetal bovine serum and saline

[0026] Figure 6 The stability of radioactive probe 3 in fetal bovine serum and saline

[0027] Figure 7 This is a 1-hour uptake and blocking experiment of three radioactive probes in Caki-1 cells;

[0028] Figure 8 Micro-PET / CT imaging at different time points after injection of radioactive probe 1 in the Caki-1-bearing mouse model;

[0029] Figure 9 Micro-PET / CT imaging at different time points after injection of radioactive probe 2 in the Caki-1-bearing mouse model;

[0030] Figure 10 Micro-PET / CT imaging at different time points after injection of radioactive probe 3 in the Caki-1-bearing mouse model;

[0031] Figure 11 For Micro-PET / CT quantitative analysis of the distribution of radioactive probe 1 in various organs and tumors at different time points in Caki-1-bearing mice;

[0032] Figure 12 Micro-PET / CT quantitative analysis of the distribution of radioactive probe 2 in various organs and tumors at different time points in Caki-1-bearing mice;

[0033] Figure 13 Micro-PET / CT quantitative analysis of the distribution of radioactive probe 3 in various organs and tumors at different time points in Caki-1-bearing mice. DETAILED DESCRIPTION

[0034] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0035] Example

[0036] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 1 is as follows:

[0037] NaH (2.5 g, 0.10 mol) was dissolved in THF (80 mL), and compound 2 (10 g, 0.04 mol) was added at room temperature. After stirring for 1 h, compound 1 was added and stirred at room temperature overnight. The reaction was complete as indicated by plate monitoring (DCM:MeOH = 20:1). 20 ml of water was added, and the mixture was extracted three times with EA. The EA was spin-dried and purified by column chromatography (DCM:MeOH system) to obtain 88 g of an orange-red oil. 1 H NMR (500 MHz, Chloroform- d ) δ 8.24 (t, J = 5.5 Hz, 1H), 8.05 (dd, J = 7.5, 1.6Hz, 1H), 7.57 (td, J = 7.5, 1.5 Hz, 1H), 7.20 (dd, J = 7.5, 1.5 Hz, 1H), 7.13(td, J = 7.5, 1.5 Hz, 1H), 3.43 (td, J = 7.1, 5.5 Hz, 2H), 3.35 (t, J = 7.0Hz, 4H), 2.54 (td, J = 7.2, 5.9 Hz, 6H), 2.01 – 1.83 (m, 2H), 1.46 (s, 7H).

[0038]

[0039] Compound 3 (8.8 g, 0.024 mol), iron powder (8.0 g, 0.143 mol), and ammonium chloride (2.5 g, 0.047 mol) were dissolved in EtOH:H2O (45 mL). After reacting at 78°C for 5 hours, a spot plate monitor (DCM:MeOH = 20:1) indicated the reaction was complete. The iron powder was filtered, 20 mL of water was added, and the mixture was extracted three times with EA. The EA was then spin-dried and purified by column chromatography (DCM:MeOH system) to yield 6.2 g of a black solid. 1 H NMR (500 MHz, Chloroform- d ) δ 7.00 (td, J = 7.5, 1.5 Hz, 1H), 6.76(ddd, J = 15.3, 7.6, 1.4 Hz, 2H), 6.60 (dd, J= 7.5, 1.5 Hz, 1H), 5.67 (t, J = 5.9 Hz, 1H), 4.95 (s, 1H), 3.39 (t, J = 7.1 Hz, 2H), 3.38 – 3.29 (m, 3H), 3.32 – 3.22 (m, 1H), 2.52 (dt, J = 10.3, 7.1 Hz, 6H), 2.03 – 1.85 (m, 2H), 1.46 (s, 6H).

[0040]

[0041] Compound 4 (6.0 g, 0.018 mol) was dissolved in THF and incubated at 0°C for 5 minutes. Compound 5 (1.7 ml, 0.018 mol) was then added and stirred for 5 hours. A microplate reader indicated the reaction was complete. The solvent was evaporated and the product was purified by column chromatography (DCM:MeOH) to yield 420 mg of a pink-yellow solid. 1 H NMR (500 MHz, Chloroform- d ) δ 9.41 (s, 1H), 7.50(dd, J = 7.4, 1.5 Hz, 1H), 7.12 (td, J = 7.4, 1.5 Hz, 1H), 6.98 (td, J = 7.5,1.6 Hz, 1H), 6.94 (t, J = 5.8 Hz, 1H), 6.84 (dd, J = 7.4, 1.5 Hz, 1H), 3.83(t, J = 7.1 Hz, 2H), 3.33 (t, J = 7.0 Hz, 4H), 3.27 (td, J = 7.1, 5.8 Hz,2H), 2.81 (t, J = 7.1 Hz, 2H), 2.59 (t, J = 7.1 Hz, 2H), 2.54 (dt, J = 14.1,7.1 Hz, 4H), 1.94 (p, J = 7.1 Hz, 2H), 1.46 (s, 7H).

[0042]

[0043] Compound 6 (5.4 g, 0.013 mol) and cesium carbonate (6.2 g, 0.019 mol) were dissolved in DMF (60 mL) and stirred overnight at room temperature. The reaction was monitored for completion using a plate with a DCM:MeOH ratio of 20:1. Water (20 mL) was added, and the mixture was extracted three times with EA. The EA was then evaporated and purified by column chromatography (DCM:MeOH system) to yield 3 g of the product. 1 H NMR (500 MHz, Chloroform- d ) δ 9.30(s, 1H), 7.55 (dd, J = 7.0, 2.0 Hz, 1H), 7.13 – 7.03 (m, 2H), 7.01 (dd, J =6.9, 2.1 Hz, 1H), 3.73 (t, J = 7.1 Hz, 2H), 3.33 (dt, J = 9.0, 7.0 Hz, 6H),3.25 (t, J = 7.1 Hz, 2H), 2.58 (t, J = 7.1 Hz, 2H), 2.54 (t, J = 7.1 Hz, 4H),1.78 (p, J = 7.1 Hz, 2H), 1.47 (s, 7H).

[0044]

[0045] Compound 7 (1.5 g, 0.004 mol) was dissolved in THF (20 mL). Lithium aluminum hydride (0.8 g, 0.021 mol) was added to the solution in an ice bath. The reaction mixture was heated at 65°C for 20 minutes. TLC indicated complete reaction (DCM:MeOH = 20:1). The reaction mixture was cooled, and water (20 mL) was slowly added. The mixture was stirred for 1 hour, filtered, and purified by column chromatography (DCM:MeOH) to yield 1.2 g of a pale yellow oil. 1 H NMR (500 MHz, Chloroform- d ) δ 6.90 – 6.83 (m, 2H), 6.79 – 6.71 (m,1H), 6.62 (dd, J = 7.2, 1.1 Hz, 1H), 5.00 (t, J= 6.0 Hz, 1H), 3.35 – 3.23(m, 10H), 2.56 (dt, J = 21.5, 7.0 Hz, 6H), 2.05 (p, J = 7.1 Hz, 2H), 1.78 (p, J = 7.1 Hz, 2H), 1.47 (s, 7H).

[0046]

[0047] Compound 8 (0.94 g, 0.003 mol) and compound 9 (0.51 g, 0.003 mol) were dissolved in DMF (7 mL). Triethylamine (0.7 mL, 0.005 mol) was then added and stirred at room temperature overnight. TLC (DCM:MeOH = 20:1) indicated complete reaction. Water (20 mL) was added to quench the reaction, and the product was extracted three times with DCM. The DCM was then spin-dried and purified by column chromatography (DCM:MeOH system) to yield 750 mg of a pale yellow oil (Compound 10). 1 H NMR (500 MHz, Chloroform- d ) δ 8.28 –8.22 (m, 1H), 8.03 – 7.97 (m, 1H), 7.07 – 6.93 (m, 2H), 3.86 (t, J = 7.1 Hz,1H), 3.31 (tt, J = 9.5, 7.1 Hz, 4H), 2.57 (q, J = 7.3 Hz, 2H), 2.50 (t, J =7.1 Hz, 1H), 1.97 (p, J = 7.1 Hz, 1H), 1.79 (p, J = 7.1 Hz, 1H), 1.45 (s,3H).

[0048]

[0049] Compound 10 (750 mg, 1.432 mmol), iron powder (480 mg, 8.594 mmol), and ammonium chloride (153 mg, 2.860 mmol) were dissolved in EtOH:H2O (12 mL). After reacting at 78°C for 3 hours, TLC (DCM:MeOH = 20:1) indicated completion of the reaction. The iron powder was filtered, 20 mL of water was added, and the mixture was extracted three times with EA. The EA was then spin-dried and purified by column chromatography (DCM:MeOH) to yield 545 mg of the compound as a pale yellow oil. 1 H NMR (500 MHz, Chloroform- d ) δ 7.54 – 7.49 (m,2H), 7.06 – 6.93 (m, 5H), 6.67 – 6.61 (m, 2H), 5.19 (d, J = 5.7 Hz, 1H), 4.98(d, J = 5.7 Hz, 1H), 3.86 (t, J = 7.1 Hz, 2H), 3.31 (tt, J = 8.2, 7.1 Hz,8H), 2.57 (td, J = 7.1, 5.6 Hz, 4H), 2.51 (t, J = 7.0 Hz, 2H), 1.97 (p, J =7.1 Hz, 2H), 1.80 (p, J = 7.1 Hz, 2H), 1.44 (s, 7H).

[0050]

[0051] Compound 11 (500 mg, 1.014 mmol), compound 12 (280 mg, 1.414 mmol), and HATU (540 mg, 1.421 mmol) were dissolved in DCM (5 mL). Triethylamine (310 μl, 2.234 mmol) was then added. The mixture was stirred at room temperature overnight. TLC (DCM:MeOH = 15:1) indicated complete reaction. Column purification (DCM:MeOH) afforded 270 mg of a pale white foamy solid (compound 13). 1 H NMR (500 MHz, Chloroform- d) δ 9.16 (s, 1H), 7.95 –7.92 (m, 2H), 7.71 – 7.66 (m, 3H), 7.63 – 7.59 (m, 1H), 7.58 – 7.53 (m, 2H),7.49 – 7.44 (m, 2H), 7.43 – 7.35 (m, 3H), 7.05 (ddd, J = 7.3, 5.9, 3.3 Hz,1H), 7.00 – 6.95 (m, 3H), 3.86 (t, J = 7.1 Hz, 2H), 3.35 – 3.27 (m, 8H), 2.57(t, J = 7.0 Hz, 2H), 2.50 (t, J = 7.1 Hz, 4H), 1.97 (p, J = 7.1 Hz, 2H), 1.76(p, J = 7.1 Hz, 2H), 1.44 (s, 9H).

[0052]

[0053] Compound 13 (50 mg, 0.074 mmol) was dissolved in DCM (200 μl) and trifluoroacetic acid (250 μl, 3.289 mmol) was added. After stirring at room temperature for 1 hour, TLC (DCM:MeOH = 10:1) indicated that the starting material had essentially reacted. The solvent was evaporated and the mixture was scraped off with a large plate (DCM:EA = 10:1) to afford 38 mg of a milky white solid (compound 14). 1 H NMR (500 MHz, Chloroform- d ) δ 9.18 (s, 1H), 7.98 – 7.95 (m, 2H), 7.73 – 7.68 (m, 3H), 7.62 (td, J =7.4, 1.6 Hz, 1H), 7.56 (dd, J = 7.5, 1.6 Hz, 1H), 7.51 (td, J = 7.3, 1.6 Hz,1H), 7.49 – 7.42 (m, 4H), 7.41 – 7.36 (m, 1H), 7.02 (dtd, J = 20.9, 7.4, 1.7Hz, 2H), 6.95 (ddd, J= 7.3, 4.7, 1.7 Hz, 2H), 3.86 (t, J = 7.1 Hz, 2H), 3.31(dt, J = 17.6, 7.1 Hz, 4H), 2.76 – 2.71 (m, 4H), 2.64 – 2.51 (m, 6H), 2.05 –1.93 (m, 3H), 1.79 (p, J = 7.1 Hz, 2H).

[0054]

[0055] Compound 14 (15 mg, 0.026 mmol) and compound 15 (16 mg, 0.035 mmol) were dissolved in DMSO (300 μl), and triethylamine (5 ml, 0.036 mmol) was added. The reaction was stirred at room temperature overnight. LC-MS showed product elution at 7.5 min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) to yield 12 mg of a white solid after lyophilization. 1 HNMR (500 MHz, Chloroform- d ) δ 9.16 (s, 1H), 7.95 – 7.89 (m, 2H), 7.73 – 7.64 (m, 3H), 7.61 (td, J = 7.3, 1.7 Hz, 1H), 7.58 – 7.50 (m, 2H), 7.48 – 7.41 (m,2H), 7.41 – 7.33 (m, 3H), 7.10 – 7.03 (m, 1H), 7.01 – 6.92 (m, 3H), 3.85 (td, J = 7.0, 5.0 Hz, 2H), 3.56 – 3.47 (m, 6H), 3.45 (s, 2H), 3.39 – 3.25 (m, 7H), 2.80 – 2.68 (m, 11H), 2.66 – 2.60 (m, 1H), 2.63 – 2.51 (m, 8H), 2.54 – 2.46 (m, 4H), 2.32 (dt, J = 12.5, 7.1 Hz, 1H), 2.24 (dt, J = 12.5, 7.1 Hz, 1H), 2.04 – 1.91 (m, 3H), 1.84 – 1.68 (m, 3H).

[0056]

[0057] Compound 16 (20 μg, 0.019 μmol) and 1 M sodium acetate (35 μl) were mixed and then 200 μl of 0.1% HCl (containing 68 GaCl31.2mCi), heated to 95℃ and heated for 10min, HPLC showed (30%A70%B, A: acetonitrile, B: 0.1% trifluoroacetic acid) the labeling rate was greater than 95%, no purification was required, such as Figure 1 As shown, compound 17 is the radioactive probe 1.

[0058] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 2 is as follows:

[0059]

[0060] Compound 18 (5 g, 0.027 mol), compound 19 (8.1 g, 0.040 mol), and potassium carbonate (5.5 g, 0.040 mol) were dissolved in DMF (60 mL). After reacting at 60°C for 30 min, TLC (DCM:MeOH = 20:1) indicated completion of the reaction. 20 mL of water was added, and the mixture was extracted three times with EA. The EA was then evaporated, and the mixture was purified by column chromatography (DCM:MeOH system) to afford compound 20. 1 H NMR (500 MHz, Chloroform- d ) δ 3.40 (t, J = 7.1 Hz, 1H), 3.35 (t, J = 7.1 Hz, 2H),2.61 (t, J = 7.0 Hz, 1H), 2.51 (t, J = 7.1 Hz, 2H), 1.95 (p, J = 7.1 Hz, 1H),1.46 (s, 3H).

[0061]

[0062] NaF (0.89 g, 0.037 mol) was dissolved in DMF (15 ml), and compound 21 (2 g, 0.004 mol) was added. The mixture was stirred for 1 hour, then incubated at 0°C and compound 20 (1.3 g, 0.004 mol) was added. The mixture was stirred overnight at room temperature. TLC (DCM:MeOH = 10:1) indicated completion of the reaction. 20 ml of water was added, and the mixture was extracted three times with EA. The EA was then evaporated, and the mixture was purified by column chromatography (DCM:MeOH system) to obtain 2 g of a pale yellow foamy solid (compound 22). 1 H NMR (500 MHz, Chloroform- d ) δ 9.15 (s, 1H), 7.71 – 7.65 (m, 2H), 7.54 – 7.52 (m, 1H), 7.41 (dd, J = 7.5, 1.4 Hz, 2H), 7.38 – 7.25 (m,5H), 4.75 (td, J = 6.7, 0.8 Hz, 1H), 3.84 (t, J = 6.9 Hz, 2H), 3.54 (dt, J =12.3, 7.1 Hz, 1H), 3.46 (dt, J = 12.4, 7.1 Hz, 1H), 3.31 (td, J = 7.1, 3.3Hz, 4H), 2.71 – 2.58 (m, 2H), 2.54 – 2.41 (m, 10H), 1.97 – 1.87 (m, 4H), 1.80(p, J = 7.1 Hz, 2H), 1.44 (s, 9H).

[0063]

[0064] Compound 22 (210 mg, 0.311 mmol) was dissolved in DCM (5 mL) and trifluoroacetic acid (1 mL, 0.013 mmol) was added. After stirring overnight at room temperature, TLC (DCM:MeOH = 10:1) indicated that the starting material had reacted almost completely. The solvent was evaporated and the product was purified by column chromatography (DCM:MeOH) to afford 164 mg of the product (compound 23) as a pale yellow foam. 1 H NMR (500 MHz, Chloroform- d ) δ 9.15 (s, 1H), 7.76 (dd, J= 7.2, 1.8 Hz, 1H), 7.70 (d, J =7.5 Hz, 1H), 7.57 – 7.53 (m, 1H), 7.42 (dd, J = 7.5, 1.7 Hz, 1H), 7.40 – 7.24(m, 6H), 4.79 – 4.73 (m, 1H), 3.87 – 3.80 (m, 2H), 3.55 (dt, J = 12.5, 7.1Hz, 1H), 3.48 (dt, J = 12.5, 7.1 Hz, 1H), 2.74 (q, J = 7.1 Hz, 4H), 2.60(ddtd, J = 28.4, 14.1, 7.1, 5.2 Hz, 6H), 2.46 – 2.41 (m, 5H), 2.02 (p, J =5.2 Hz, 1H), 1.99 – 1.87 (m, 4H), 1.81 (pd, J = 7.2, 2.0 Hz, 2H).

[0065]

[0066] Compound 23 (7.7 mg, 0.013 mmol) and compound 15 (6.8 mg, 0.151 mmol) were dissolved in DMSO (200 μl). Triethylamine (2.06 μl, 0.015 mmol) was then added. The reaction was stirred at room temperature overnight. LC-MS showed product elution at 7.6 min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) to yield 3 mg of a white solid after lyophilization. 1 HNMR (500 MHz, Chloroform- d ) δ 9.15 (s, 1H), 7.71 – 7.65 (m, 2H), 7.56 – 7.53(m, 1H), 7.40 – 7.25 (m, 7H), 4.78 – 4.73 (m, 1H), 3.82 (td, J= 6.9, 1.1 Hz,2H), 3.57 – 3.43 (m, 10H), 3.38 – 3.32 (m, 3H), 2.79 – 2.69 (m, 10H), 2.69 –2.48 (m, 12H), 2.45 – 2.41 (m, 6H), 2.32 (dt, J = 12.5, 7.1 Hz, 1H), 2.24(dt, J = 12.5, 7.1 Hz, 1H), 2.03 – 1.86 (m, 5H), 1.84 – 1.74 (m, 3H).

[0067]

[0068] Compound 24 (20 μg, 0.019 μmol) and 1 M sodium acetate (35 μl) were mixed and then 200 μl of 0.1% HCl (containing 68 GaCl31.2mCi), heated to 95℃ and heated for 10min, HPLC showed (30%A70%B, A: acetonitrile, B: 0.1% trifluoroacetic acid) the labeling rate was greater than 95%, no purification was required, such as Figure 2 As shown, compound 25 is the radioactive probe 2.

[0069] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 3 is as follows:

[0070]

[0071] Compound 18 (2.8 g, 0.015 mol), compound 26 (5.0 g, 0.018 mol), and potassium carbonate (2.5 g, 0.018 mol) were dissolved in DMF (40 mL) and stirred at room temperature. TLC (DCM:MeOH = 10:1) indicated completion of the reaction. 20 mL of water was added, and the mixture was extracted three times with EA. The EA was then evaporated and purified by column chromatography (DCM:MeOH system) to afford 3 g of a colorless oil (Compound 27). 1 H NMR (500 MHz, Chloroform- d ) δ 3.93 (t, J = 7.1 Hz, 2H), 3.68– 3.59 (m, 8H), 3.33 (t, J = 7.1 Hz, 4H), 3.12 (dt, J= 12.6, 7.1 Hz, 1H),2.76 (dt, J = 12.8, 7.0 Hz, 1H), 2.53 (t, J = 7.1 Hz, 4H), 1.46 (s, 7H).

[0072]

[0073] Compound 21 (973.3 mg, 2.168 mmol), compound 27 (829.1 mg, 2.174 mmol), and cesium carbonate (1419.9 mg, 4.358 mmol) were dissolved in DMF (24 ml) and heated with stirring at 60°C overnight. TLC indicated a 50% reaction (DCM:MeOH = 10:1). Add 50 ml of water, extract with EA three times, spin-dry the EA, and purify by column chromatography (DCM:MeOH system) to obtain compound 28. 1 HNMR (500 MHz, Chloroform- d ) δ 9.15 (s, 1H), 7.71 (dd, J = 7.2, 1.8 Hz, 1H),7.66 (d, J = 7.5 Hz, 1H), 7.55 – 7.51 (m, 1H), 7.41 – 7.24 (m, 7H), 4.78 (t, J = 6.7 Hz, 1H), 3.82 (td, J = 6.9, 1.3 Hz, 2H), 3.79 – 3.69 (m, 2H), 3.68 –3.51 (m, 8H), 3.31 (dt, J = 8.4, 7.1 Hz, 4H), 3.05 (dt, J = 12.4, 7.0 Hz,1H), 2.76 (dt, J = 12.4, 7.0 Hz, 1H), 2.53 (t, J = 7.1 Hz, 2H), 2.50 – 2.43(m, 5H), 2.42 (d, J = 0.7 Hz, 3H), 1.99 – 1.85 (m, 4H), 1.44 (s, 7H).

[0074]

[0075] Compound 28 (264.9 mg, 0.354 mmol) was dissolved in DCM (1 ml). Trifluoroacetic acid (1 ml, 0.013 mmol) was added and stirred overnight at room temperature. TLC (DCM:MeOH = 10:1) indicated that the starting material had essentially reacted. The solvent was evaporated and the product was purified by column chromatography (DCM:MeOH system) to afford compound 29. 1 H NMR (500 MHz, Chloroform- d ) δ 9.16 (s,1H), 7.71 (dd, J = 7.2, 1.7 Hz, 1H), 7.66 (d, J = 7.5 Hz, 1H), 7.55 – 7.51(m, 1H), 7.42 (dd, J = 7.5, 1.5 Hz, 1H), 7.39 – 7.24 (m, 6H), 4.81 – 4.75 (m,1H), 3.84 (t, J = 6.9 Hz, 2H), 3.81 – 3.71 (m, 1H), 3.74 – 3.64 (m, 1H), 3.67– 3.53 (m, 8H), 3.02 (dt, J = 12.5, 7.1 Hz, 1H), 2.81 (dt, J = 12.8, 7.1 Hz,4H), 2.73 (dt, J = 12.4, 7.0 Hz, 1H), 2.57 (dtd, J = 12.4, 7.1, 5.2 Hz, 4H),2.46 – 2.40 (m, 5H), 2.05 (p, J = 5.2 Hz, 1H), 1.99 – 1.84 (m, 4H).

[0076]

[0077] Compound 29 (26.6 mg, 0.024 mmol) and compound 15 (20.2 mg, 0.044 mmol) were dissolved in DMSO (500 μl). Triethylamine (115.9 μl, 0.853 mmol) was then added. The reaction was stirred at room temperature overnight. LC-MS showed product elution at 7.8 min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) to yield 3 mg of a white solid after lyophilization. 1HNMR (500 MHz, Chloroform- d ) δ 9.15 (s, 1H), 7.72 – 7.65 (m, 2H), 7.56 – 7.54(m, 1H), 7.40 – 7.25 (m, 7H), 4.78 (t, J = 6.7 Hz, 1H), 3.87 – 3.70 (m, 4H), 3.66 – 3.54 (m, 8H), 3.50 (dt, J = 10.4, 7.0 Hz, 5H), 3.45 (d, J = 2.1 Hz,6H), 3.34 (t, J = 7.0 Hz, 1H), 3.05 (dt, J = 12.3, 7.0 Hz, 1H), 2.79 – 2.69(m, 12H), 2.68 – 2.46 (m, 11H), 2.45 – 2.40 (m, 6H), 2.36 – 2.20 (m, 2H), 2.03 – 1.84 (m, 5H), 1.77 (dq, J = 12.5, 7.1 Hz, 1H).

[0078]

[0079] Compound 30 (20 μg, 0.018 μmol) and 1 M sodium acetate (35 μl) were mixed and then 200 μl of 0.1% HCl (containing 68 GaCl31.2mCi), heated to 95℃ and heated for 10min, HPLC showed (30%A70%B, A: acetonitrile, B: 0.1% trifluoroacetic acid) the labeling rate was greater than 95%, no purification was required, such as Figure 3 As shown, compound 31 is the radioactive probe 3.

[0080] The stability of the three probes was tested in fetal bovine serum and normal saline. After the three probes were synthesized, they were mixed with equal volumes of fetal bovine serum and normal saline, incubated at 37 degrees. 1-4 μl of the mixture was dropped onto a silica gel plate at 0 h, 1 h, 2 h, and 4 h, using 0.1 M citric acid as the developing agent. When the plate was run to 11 cm from the origin, the radiochemical purity was detected by radioactive thin-layer chromatography. The results were as follows: Figure 4 、 Figure 5 and Figure 6As shown, it can be seen that the radiochemical purity of the three radioactive probes in fetal bovine serum and normal saline is still greater than 95% after 4 hours, indicating that the three radioactive probes have good in vitro stability.

[0081] The targeting of the three probes was tested on Caki-1 cells. Caki-1 cells were seeded in a six-well plate and incubated with the three probes for 1 hour. The radioactivity counts of the cell supernatant and cell lysate were measured using a gamma counter to calculate the cell uptake value. At the same time, under the same conditions, three labeled precursors (i.e., compound 16, compound 24, and compound 30) were used to incubate for half an hour in advance for blocking to detect whether the three probes targeted V2R. The results are shown in Figure 2. Figure 7 As shown in the figure, the blocking rates of probe 1, probe 2 and probe 3 were 18.3%, 36.4% and 30.8%, respectively, indicating that all three probes have the ability to target V2R, among which probe 2 has the best targeting ability.

[0082] PET Imaging Experiment in a Caki-1 Tumor-Bearing Mouse Model: After obtaining the radiolabeled material through positron emission tomography (PET) labeling, a sterile injection solution of the PET probe was obtained for animal imaging without purification. The Caki-1 tumor-bearing mouse model was successfully established. After anesthesia with an isoflurane-oxygen mixture using a small animal anesthesia machine, static micro-PET / CT (IRIS Micro-PET / CT, INVISCAN) scans were performed at 3.5-4.5 MBq per mouse. PET signals were acquired and reconstructed at 30, 60, and 120 minutes after injection.

[0083] The Caki-1 tumor-bearing mouse model was injected with probe 1, probe 2, and probe 3, and micro-PET / CT imaging was performed at different time points. Figure 8 、 Figure 9 and Figure 10 As shown in the figure, we can see that all three probes are retained in the tumor to varying degrees. To quantify the uptake of radioactive drugs in the body, we use the Standard Uptake Value (SUV) to evaluate drug uptake. SUV = radioactivity concentration in the lesion (kBq / ml) / injected dose (MBq) / body weight (kg). The higher the value, the higher the concentration of radioactive probe in that area. Figure 11 、 Figure 12 and Figure 13 In this experiment, the heart, liver, spleen, lungs, kidneys, stomach, small intestine, muscle, right humerus, brain, and tumor were delineated as regions of interest (ROIs). SUVs were calculated using software. This figure shows that probes 1 and 2 are primarily excreted through the liver, while probe 3 is primarily excreted through the kidneys. Probe 1 is better retained in the tumor, while probes 2 and 3 are excreted more rapidly.

[0084] It can be seen from the above examples that the probe provided by the present invention can be used to target V2R receptors for imaging, providing strong evidence for V2R-targeted therapy.

[0085] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A benzoazepine derivative, characterized in that: The benzoazepine derivative is a compound having a structure shown in Formula 1: Formula 1.

2. A benzoazepine derivative, characterized in that: The benzoazepine derivative is a compound having a structure shown in Formula 2: Formula 2.

3. A benzoazepine derivative, characterized in that: The benzoazepine derivative is a compound having a structure shown in Formula 3: Formula 3.

4. A radioactive probe, characterized in that The radioactive probe is a compound having a structure shown in Formula 4: Formula 4.

5. A radioactive probe, characterized in that The radioactive probe is a compound having a structure shown in Formula 5: Formula 5.

6. A radioactive probe, characterized in that The radioactive probe is a compound having a structure shown in Formula 6: Formula 6.

7. Use of the benzazepine derivative according to any one of claims 1 to 3 or the radioactive probe according to any one of claims 4 to 6 in the preparation of a drug targeting arginine vasopressin receptor 2.

8. Use of the benzoazepine derivative according to any one of claims 1 to 3 or the radioactive probe according to any one of claims 4 to 6 in the preparation of a cancer cell imaging agent, a cancer auxiliary diagnostic reagent, a cancer staging assessment reagent, or a cancer drug efficacy evaluation product.

9. The use according to claim 8, characterized in that The cancer is renal cell carcinoma.

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