Benzazepine derivative, radioactive probe and application thereof

Abstract

The invention provides a benzene azepine derivative, a radioactive probe and application thereof, and belongs to the technical field of biological medicine. According to the present invention, the benzazepine derivative is modified, and the hydrophilic or lipophilic side chain is introduced, such that the prepared benzazepine derivative can be used as the V2R targeting precursor for application, the specific V2R targeting can be achieved, and the new molecular tool is provided for the subsequent V2R positive tumor diagnosis and treatment. The radioactive probe combined with the 68Ga marker shows a good tumor imaging effect, can realize PET imaging of V2R positive tumors, is helpful for improving the accuracy of tumor diagnosis, can also be used for evaluating the treatment effect, has a relatively high target / non-target ratio, can better display the distribution of whole body V2R receptors, and has a good application prospect. The invention has various applications in cancer diagnosis and treatment.

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 more than 90% of renal cancers, of which clear cell renal cell carcinoma (ccRCC) accounts for the vast majority of RCC cases (70%), papillary RCC (pRCC) accounts for 10-15%, and chromophobe RCC (rhRCC) accounts for about 5%. Most RCCs are diagnosed when the lesions are confined to the kidneys. These patients have a good prognosis, with a 5-year survival rate of more than 90%. The prognosis of advanced patients is poor, such as when tumor cells have spread to surrounding tissues or organs and / or regional lymph nodes, with a 5-year survival rate of about 70%; the 5-year survival rate of patients with distant metastasis is only 13%. Although most RCCs can be cured by surgery, 20% to 30% of patients will experience distant metastasis, and 2% to 5% of patients will experience local recurrence. Therefore, the diagnosis and treatment of metastatic and recurrent RCC has always been the focus of RCC.

[0003] In the past 20 years, with the implementation of personalized medicine and precision medicine, radioactive drugs and fusion imaging equipment have been vigorously developed, and nuclear medicine has played an important role in the diagnosis and treatment of many diseases. Functional PET / CT function has been proven to be very important in the diagnosis, staging and treatment guidance of lymphoma, esophageal cancer, nasopharyngeal carcinoma, lung cancer, gastric cancer, intestinal cancer and other malignant tumors, but its application in RCC is still relatively limited. Compared with traditional imaging methods, the supplementary value of FDG PET / CT in evaluating the metastasis and recurrence of RCC is limited, and in low-grade renal cancer, the level of glucose metabolism in the lesions is often not high, and the lesions have poor ability to absorb FDG, which cannot detect tumors well. In addition, current research on RCC PET imaging agents includes 89Zr-bevacizumab, 89Zr-glenizumab, 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-glenizumab, the optimal imaging effect is usually achieved 3-7 days after injection, and there are certain limitations in clinical application. PSMA is mainly expressed in the endothelial cells of RCC new capillaries and cannot directly display the activity of tumor cells, 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 directions in nuclear medicine technology. Integrated nuclear medicine diagnosis and treatment combines imaging diagnosis with internal irradiation therapy by using different diagnostic and therapeutic radionuclide probes, thereby achieving the purpose of visual diagnosis and precise treatment. It runs through the entire process of personalized medicine, covering diagnosis, grading and staging, treatment, efficacy monitoring and prognosis judgment, so it has broad application prospects in the field of tumors. Currently, FDA-approved integrated diagnosis and treatment radiopharmaceuticals can be used for differentiated thyroid cancer, neuroendocrine tumors and prostate cancer, but there is no corresponding radiotherapy for RCC, a cancer with a high incidence and mortality rate, that has entered clinical research. Metastatic RCC is mainly treated with systemic treatments, such as tyrosine kinase inhibitors and immunotherapy, but for drug-resistant metastatic RCC, there are few alternative treatment options, and the efficacy and safety are uncertain.

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

[0006] The purpose of the present invention is to provide a benzazine derivative, a radioactive probe and its application, which can be used for in vivo tumor imaging based on V2R targets in the human body, and to detect the therapeutic efficacy of targeting V2R, so as to provide 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: The present invention provides a benzoazepine derivative, wherein the benzoazepine derivative is a compound having a structure shown in Formula 1: Formula 1.

[0008] The present invention also provides another benzoazepine derivative, wherein the benzoazepine derivative is a compound having a structure shown in Formula 2: Formula 2.

[0009] The present invention also provides another benzoazepine derivative, wherein the benzoazepine derivative is a compound having a structure shown in Formula 3: Formula 3.

[0010] The present invention also provides a radioactive probe, which is a compound having a structure shown in Formula 4: Formula 4.

[0011] The present invention also provides another radioactive probe, which is a compound having a structure shown in Formula 5: Formula 5.

[0012] The present invention also provides another radioactive probe, which is a compound having a structure shown in Formula 6: Formula 6.

[0013] The present invention also provides the use of the above-mentioned benzazepine derivatives or radioactive probes in the preparation of arginine vasopressin receptor 2 targeted drugs.

[0014] 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.

[0015] Preferably, the cancer is renal cell carcinoma.

[0016] Beneficial effects of the present invention: The present invention modifies the benzazepine derivatives and introduces hydrophilic or lipophilic side chains. The prepared benzazepine derivatives can be used as V2R-targeting precursors, can achieve specific targeting of V2R, and provide new molecular tools for subsequent diagnosis and treatment of V2R-positive tumors. The preparation method of the benzazepine derivatives of the present invention is simple, the raw materials are easily available, and have good clinical transformation prospects.

[0017] The radioactive probe of the present invention is combined with 68 Ga labeling shows good tumor imaging effect, 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 higher "target / non-target" ratio, and can better display the distribution of V2R receptors throughout the body.

[0018] The present invention provides a benzazepine derivative and 68 Ga markers have many applications in the diagnosis and treatment of cancer, including the ability to reflect the expression level of V2R in renal cell carcinoma, which can be used for the diagnosis and staging of renal cell carcinoma, as well as the formulation of treatment plans, to achieve integrated targeted V2R diagnosis and treatment, and have high clinical translational value, providing a new approach for personalized diagnosis and precision treatment of tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 To determine the labeling rate of radioactive probe 1 using radioactive high performance liquid chromatography (HPLC) Figure 2 To determine the labeling rate of radioactive probe 2 using radioactive high performance liquid chromatography (HPLC) Figure 3 To determine the labeling rate of radioactive probe 3 using radioactive high performance liquid chromatography (HPLC) Figure 4 Stability of radioactive probe 1 in fetal bovine serum and saline Figure 5Stability of radioactive probe 2 in fetal bovine serum and saline Figure 6 Stability of radioactive probe 3 in fetal bovine serum and saline Figure 7 This is a 1-hour uptake and blocking experiment of three radioactive probes by Caki-1 cells; Figure 8 Micro-PET / CT imaging at different time points after injection of radioactive probe 1 in the Caki-1-bearing mouse model; Fig. 9 Micro-PET / CT imaging at different time points after injection of radioactive probe 2 in the Caki-1-bearing mouse model; Fig.10 Micro-PET / CT imaging at different time points after injection of radioactive probe 3 in the Caki-1-bearing mouse model; Fig.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; Fig.12 For 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; Fig.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

[0020] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0021] Example

[0022] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 1 is as follows:

[0023] NaH (2.5 g, 0.10 mol) was dissolved in THF (80 mL), and compound 2 (10 g, 0.04 mol) was added thereto at room temperature. After stirring for 1 h, compound 1 was added and stirred at room temperature overnight. The spot plate monitoring (DCM:MeOH = 20:1) showed that the reaction was complete. 20 ml of water was added, and EA was extracted 3 times. The EA was spin-dried and purified by column (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).

[0024] 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, the DCM:MeOH=20:1 spot plate monitoring indicated that the reaction was complete. The iron powder was filtered, 20 ml of water was added, EA was extracted 3 times, EA was spin-dried, and column purification (DCM: MeOH system) was performed to obtain 6.2 g of 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).

[0025] Compound 4 (6.0 g, 0.018 mol) was dissolved in THF and placed at 0°C for 5 minutes, then compound 5 (1.7 ml, 0.018 mol) was added and stirred for 5 hours. The plate monitoring indicated that the reaction was complete. The solvent was spin-dried and purified by column (DCM: MeOH system) to obtain 420 mg of 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).

[0026] Compound 6 (5.4 g, 0.013 mol) and cesium carbonate (6.2 g, 0.019 mol) were mixed and dissolved in DMF (60 mL), stirred at room temperature overnight, and the reaction was monitored by a plate with DCM:MeOH=20:1. Water (20 mL) was added, and EA was extracted three times. The EA was dried by spin drying, and the product was purified by column purification (DCM: MeOH system) to obtain 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).

[0027] Compound 7 (1.5 g, 0.004 mol) was dissolved in THF (20 mL), placed in an ice bath, and lithium aluminum hydride (0.8 g, 0.021 mol) was added. The mixture was heated at 65°C for 20 minutes. TLC showed (DCM: MeOH = 20: 1) that the reaction of the raw material was complete. The reaction mixture was cooled, and water (20 mL) was slowly added. After stirring for 1 hour, the mixture was filtered and purified by column (DCM: MeOH system) to obtain 1.2 g of a light 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).

[0028] Compound 8 (0.94 g, 0.003 mol) and compound 9 (0.51 g, 0.003 mol) were dissolved in DMF (7 mL), and triethylamine (0.7 ml, 0.005 mol) was added. The mixture was stirred overnight at room temperature. TLC showed (DCM: MeOH = 20: 1) that the raw materials were completely reacted. Water (20 mL) was added to quench the reaction, and DCM was extracted three times. The DCM was dried by spin drying and purified by column (DCM: MeOH system) to obtain 750 mg of a light 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).

[0029] 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) spot plate monitoring indicated that the reaction was complete. The iron powder was filtered, 20 ml of water was added, EA was extracted 3 times, EA was spin-dried, and column purification (DCM:MeOH system) was performed to obtain 545 mg of a light yellow oily compound. 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).

[0030] Compound 11 (500 mg, 1.014 mmol), compound 12 (280 mg, 1.414 mmol), HATU (540 mg, 1.421 mmol) were dissolved in DCM (5 mL), and triethylamine (310 ul, 2.234 mmol) was added. The mixture was stirred at room temperature overnight. TLC showed (DCM:MeOH=15:1) that the raw materials were completely reacted. Column purification (DCM:MeOH system) was performed to obtain 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).

[0031] Compound 13 (50 mg, 0.074 mmol) was dissolved in DCM (200 ul), trifluoroacetic acid (250 ul, 3.289 mmol) was added and stirred at room temperature for 1 h. TLC showed that (DCM: MeOH = 10: 1) the raw material was basically reacted. The solvent was spin-dried and scraped on a large plate (DCM: EA = 10: 1) to obtain 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).

[0032] Compound 14 (15 mg, 0.026 mmol) and compound 15 (16 mg, 0.035 mmol) were dissolved in DMSO (300ul), and triethylamine (5 ml, 0.036 mmol) was added. The mixture was stirred at room temperature overnight. LC-MS showed that the product peaked at 7.5min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) and lyophilized to obtain 12 mg of a white solid. 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).

[0033] Compound 16 (20ug, 0.019 umol) and 1M sodium acetate (35ul) were mixed and then 200ul 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.

[0034] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 2 is as follows:

[0035] 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 that the reaction was complete. 20 ml of water was added, EA was extracted 3 times, EA was dried by spin drying, and column purification (DCM: MeOH system) was performed to obtain 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).

[0036] NaF (0.89 g, 0.037 mol) was dissolved in DMF (15 ml), compound 21 (2 g, 0.004 mol) was added, stirred for 1 h, then placed at 0°C, compound 20 (1.3 g, 0.004 mol) was added, stirred overnight at room temperature, TLC showed (DCM: MeOH = 10:1) the reaction was complete. 20 ml of water was added, EA was extracted 3 times, EA was dried by spin drying, and column purification (DCM: MeOH system) was performed to obtain 2 g of a light 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).

[0037] Compound 22 (210 mg, 0.311 mmol) was dissolved in DCM (5 ml), trifluoroacetic acid (1 ml, 0.013 mmol) was added, and the mixture was stirred overnight at room temperature. TLC showed that (DCM: MeOH = 10:1) the reaction of the raw material was basically complete. The solvent was dried and purified by column chromatography (DCM: MeOH system) to obtain 164 mg of a light yellow foamy product (compound 23). 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).

[0038] Compound 23 (7.7 mg, 0.013 mmol) and compound 15 (6.8 mg, 0.151 mmol) were dissolved in DMSO (200 ul), and triethylamine (2.06 ul, 0.015 mmol) was added. The mixture was stirred at room temperature overnight. LC-MS showed that the product peaked at 7.6 min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) and lyophilized to obtain 3 mg of a white solid. 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).

[0039] Compound 24 (20ug, 0.019 umol) and 1M sodium acetate (35ul) were mixed and then 200ul 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.

[0040] 68 The specific synthesis method of Ga-chelated V2R small molecule inhibitor probe 3 is as follows:

[0041] 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 showed (DCM: MeOH = 10:1) that the reaction was complete. 20 ml of water was added, EA was extracted 3 times, EA was dried by spin drying, and column purification (DCM: MeOH system) was performed to obtain 3 g of 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).

[0042] 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), heated and stirred at 60°C overnight, and TLC showed (DCM:MeOH=10:1) that the reaction reached 50%. 50 ml of water was added, and EA was extracted three times. The EA was spin-dried, and the mixture was purified by column (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).

[0043] Compound 28 (264.9 mg, 0.354 mmol) was dissolved in DCM (1 ml), trifluoroacetic acid (1 ml, 0.013 mmol) was added, and the mixture was stirred overnight at room temperature. TLC showed that (DCM: MeOH = 10:1) the reaction of the raw material was basically complete. The solvent was dried and purified by column chromatography (DCM: MeOH system) to obtain 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).

[0044] Compound 29 (26.6 mg, 0.024 mmol) and compound 15 (20.2 mg, 0.044 mmol) were dissolved in DMSO (500 ul), and triethylamine (115.9 ul, 0.853 mmol) was added. The mixture was stirred at room temperature overnight. LC-MS showed that the product peaked at 7.8 min. The product was purified by HPLC (acetonitrile: 0.1% formic acid system) and lyophilized to obtain 3 mg of a white solid. 1 HNMR (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).

[0045] Compound 30 (20ug, 0.018 umol) and 1M sodium acetate (35ul) were mixed and then 200ul of 0.1% HCl (containing 68GaCl31.2mCi), 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.

[0046] The stability of the three probes was tested in fetal bovine serum and saline. After the three probes were synthesized, they were mixed with equal volumes of fetal bovine serum and saline, respectively, and incubated at 37 degrees. 1-4ul of the mixed solution was dropped onto a silica gel plate at 0h, 1h, 2h, and 4h, respectively. 0.1M citric acid was used as the developing agent. When the plate was run to 11cm from the origin, a radioactive thin layer chromatography was used to detect its radiochemical purity. The results were as follows: Figure 4 , Figure 5 and Figure 6 As shown, it can be seen that the radiochemical purity of the three radioactive probes in fetal bovine serum and physiological saline is still greater than 95% after 4 hours, indicating that the three radioactive probes have good in vitro stability.

[0047] The targeting of the above three probes was detected 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 block the cells by incubation for half an hour in advance to detect whether the three probes targeted V2R. The results are shown in Figure 2. Figure 7 As shown, 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.

[0048] PET imaging experiment of Caki-1 mouse tumor model: After obtaining the radioactive marker through the positron nuclide labeling experiment, the sterile injection solution of nuclide probe that can be used for animal imaging is obtained without purification. The Caki-1 mouse tumor model was successfully established. After anesthesia with isoflurane-oxygen mixed gas using a small animal anesthesia machine, 3.5-4.5MBq was used for each mouse, and Micro PET / CT (IRIS Micro-PET / CT, INVISCAN) static scanning was performed. PET signals were collected and PET images were reconstructed at 30, 60 and 120 minutes after injection.

[0049] 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 , Fig. 9 and Fig.10 As shown, it can be seen that the three probes are retained in the tumor to varying degrees. In order to quantify the uptake of radioactive drugs in the body, we use SUV (Standard Uptake Value) to evaluate the uptake of drugs. SUV = radioactive concentration of lesions (kBq / ml) / injection dose (MBq) / body weight (kg). The higher the value, the higher the concentration of radioactive probes in the area. Fig.11 , Fig.12 and Fig.13 In this experiment, the heart, liver, spleen, lung, kidney, stomach, small intestine, muscle, right humerus, brain, and tumor were delineated as regions of interest, and the SUV was calculated by software. It can be seen from the figure that probes 1 and 2 are mainly excreted through the liver, while probe 3 is mainly excreted through the kidneys. Probe 1 has a better retention effect in the tumor, and probes 2 and 3 are excreted faster.

[0050] 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.

[0051] 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 principle of the present invention. These improvements and modifications should also be regarded as 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 arginine vasopressin receptor 2 targeted drugs.

8. Use of the benzazine 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 cancer cell imaging agents, cancer auxiliary diagnostic reagents, cancer staging evaluation reagents or cancer drug efficacy evaluation products.

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

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