PSMA-targeted fluorescent dimer probe as well as pharmaceutical salt and application of PSMA-targeted fluorescent dimer probe

Abstract

The invention aims to provide a compound as well as a composition, a kit and application thereof, mainly relates to the fields of medicinal chemistry, molecular probes and biomedical engineering, and in particular relates to a PSMA-targeted EuK dimer compound as well as pharmaceutical salt and application thereof. The structure of the EuK dimer compound is as shown in formula (I): # imgabs0 #. The novel PSMA-targeted cyanine EuK dimer compound provided by the invention has very high affinity with a prostate specific membrane antigen, has good in-vivo pharmacokinetic properties, is suitable for early diagnosis and staging of prostate cancer, and has good application prospects. And the probe can also be used for navigation or cleaning in a fluorescence operation in a prostatic cancer resection operation. The high-affinity fluorescent probe molecules can specifically target tumors, can accurately position tumor boundaries, can provide real-time image navigation in an operation, and have the advantage of improving the accuracy of the operation.

Description

Technical Field

[0001] The present invention relates to the fields of pharmaceutical chemistry, molecular probes and biomedical engineering, and in particular to the application of PSMA-targeting EuK dimer fluorescently labeled compounds and pharmaceutical salts thereof in the diagnosis of prostate cancer and intraoperative navigation precise resection. Background Art

[0002] The treatment of prostate cancer depends on the stage of the disease at the time of initial diagnosis. For patients with early localized prostate cancer who need surgical resection, radical prostatectomy (RP) is still the main treatment method. After RP treatment, the 5-year survival rate is usually over 98%, showing a very high treatment effectiveness. However, there are two challenges in the surgical treatment process. On the one hand, how to control the positive surgical margin. Due to the invasive growth of the tumor, the tumor edge will be irregular, making it difficult to completely remove the cancerous tissue without damaging the normal tissue. Although complete resection of the prostate can ensure the complete removal of the primary lesion, it may cause damage to functional tissues such as peripheral nerves, resulting in side effects including urinary incontinence and erectile dysfunction. Clinicians need to face the balance between complete resection and preservation of nerve function and the challenge of judging the resection margin. On the other hand, when performing lymph node dissection on patients, most lymph nodes cannot be distinguished by vision and touch, and local lymph node metastasis in the pelvis cannot be identified. How doctors accurately identify and remove diseased lymph nodes during surgery is also a major challenge. Incomplete resection will greatly increase the probability of recurrence and significantly affect the prognosis.

[0003] In recent years, the PSMA fluorescence-guided surgery (FGS) with targeted capabilities has developed rapidly, providing surgeons with a new and important method to accurately determine the location and margins of tumors. When using laparoscopic systems and da Vinci robots, various fluorescent cameras can be equipped to provide real-time and effective signal guidance during surgery, helping doctors to accurately determine the margins of tumors and reduce the probability of positive resection margins. In addition, fluorescent probes are not limited by the half-life of radionuclides, providing a longer surgical time window, helping to reduce the accumulation of probe molecules in the bladder and avoiding urine contamination of the surgical field.

[0004] Fluorescent probes currently approved by the FDA for clinical use include sodium fluorescein, methylene blue (MB), indocyanine green (ICG), 5-aminolevulinic acid (5-ALA) and its hexyl ester. Although these probes have played an important role in a variety of surgeries, they have certain limitations in prostate cancer surgery. In particular, although ICG has been initially used in sentinel lymph node identification and lymph node dissection for prostate cancer, its lack of targeting and weak binding ability at the tumor site result in a short visualization surgery time window and low sensitivity to small lesions.

[0005] Prostatic Specific Membrane Antigen (PSMA) is highly expressed in prostate cancer cells, reaching 100-1000 times that of normal tissues, and is positively correlated with Gleason scores, making PSMA an important biomarker for prostate cancer and a target with great application potential in diagnosis and treatment. In actual use, the most widely studied small molecule inhibitors targeting PSMA are mainly glutamic acid urea inhibitors, including EuK (glutamic acid-urea-lysine) and EuE (glutamic acid-urea-glutamic acid).

[0006] In response to practical problems encountered during surgery, the development of near-infrared (NIR) fluorescent molecules targeting PSMA for use in fluorescence navigation during prostate cancer surgical resection has great application potential. Summary of the invention

[0007] The present invention provides a PSMA-targeted cyanine EuK dimer fluorescent probe or a pharmaceutically acceptable salt thereof. Such compounds can be used for fluorescence intraoperative navigation or sweeping in prostate cancer resection, can accurately locate the tumor boundary, can provide real-time image navigation during surgery, and have the advantage of improving surgical accuracy.

[0008] To achieve the above object, the first embodiment of the present invention provides a compound of formula I, whose structure is shown in formula (I):

[0009]

[0010] Wherein, F is the fluorophore;

[0011] W is a PSMA targeting ligand or its analogue; L1 is absent or represented by n is an integer from 1 to 12; L2 does not exist or is represented by n is an integer from 1 to 12.

[0012] F is selected from the following:

[0013]

[0014] Wherein, R1 and R2 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ;

[0015] R3 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0016] n is an integer between 1 and 5;

[0017]

[0018] Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ;

[0019] R5 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0020] n=an integer between 1 and 5;

[0021]

[0022] Wherein, R1 and R2 are simultaneously or independently represented by SO3- M + or SO3H,M + Mainly selected from Na + , K + , Li + ;

[0023] R3 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0024] R4 represents Cl or

[0025]

[0026] Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ;

[0027] R5 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0028] R6=Cl or

[0029]

[0030] Wherein, R1 and R2 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li+ ;

[0031] R3 and R4 are simultaneously or independently CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0032]

[0033] Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ;

[0034] R5 and R6 are simultaneously or independently CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ;

[0035] In some embodiments, the compound is selected from the group consisting of:

[0036]

[0037] The compound of formula I provided by the present invention has high affinity with PSMA and high tumor target / non-target ratio. It is found in the in vivo fluorescence imaging of tumor-bearing mice that it can be effectively enriched at the tumor site and can achieve long-term retention. It is mainly excreted through the kidneys and liver, and has fast metabolism and low uptake in non-target tissues and organs. It has good in vivo pharmacokinetic properties and can be accurately used in the diagnosis, detection and intraoperative navigation and precise resection of prostate cancer and related diseases.

[0038] The second aspect of the present invention provides a pharmaceutical composition comprising a compound according to any one of the embodiments of the first aspect, and optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0039] A third aspect of the present invention provides a kit comprising the compound of any embodiment of the first aspect and the pharmaceutical composition of any embodiment of the second aspect as active ingredients.

[0040] The fourth aspect of the present invention provides a reagent for diagnosing or detecting abnormal prostate cancer-specific membrane antigen (PSMA), comprising a compound of any embodiment of the first aspect and a pharmaceutical composition provided by the second aspect as active ingredients.

[0041] A fifth aspect embodiment of the present invention provides the use of the compound of any embodiment of the first aspect or the pharmaceutical composition of any embodiment of the second aspect as an active ingredient for diagnosis and / or treatment selected from the group consisting of: preparation of PSMA detection reagents, preparation of prostate cancer detection reagents, diagnosis of prostate cancer, fluorescence navigation in surgical treatment, and detection of clinical samples of prostate cancer.

[0042] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects:

[0043] The fluorescent molecular probe provided by the present invention has good water solubility and optical properties, and can be directly used as a small molecule fluorescent probe for detecting abnormal PSMA in biological tissue samples. When using a 1 nmol / mouse compound solution for in vivo imaging of tumor-bearing mice, the tumor targeting ability is strong and the imaging effect is good. It can help doctors to accurately locate the tumor boundary during surgery when using molecular imaging surgical navigation equipment to achieve accurate tumor resection, and can provide real-time fluorescence guidance during lymph node dissection to achieve identification and positioning of abnormal lymph nodes and accurate resection, thereby reducing the risk of postoperative recurrence. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0045] Figure 1 It is a synthetic route map.

[0046] Figure 2 This is a cell staining image.

[0047] Figure 3 This is the NIR-I fluorescence imaging in tumor-bearing mice.

[0048] Figure 4 This is the NIR-II fluorescence imaging in tumor-bearing mice.

[0049] Figure 5 This is a fluorescence imaging image of a surgical simulation experiment under fluorescence guidance. DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are 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 creative work are within the scope of protection of the present invention.

[0051] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0052] The synthetic route of the following example compounds is as follows Figure 1 As shown; wherein a to c represent the reaction reagents and conditions of each step, specifically: (a) iPrOH, DMTMM, 40°C; (b) CH2Cl2 / CF3COOH, rt; (c) Na2CO3 (aqueous solution), 75°C.

[0053] Example 1 Synthesis of Compound 1-1

[0054]

[0055] Compound 1-1 was synthesized by the method described in the literature (Eur J Med Chem, 2011, 46(10):5175-83). 1HNMR(400MHz,DMSO-d6)δ12.03(d,J=4.3Hz,1H),8.25(dt,J=8.7,4.0Hz,2H),8.16–7.90(m ,6H),7.89–7.74(m,2H),7.74–7.68(m,1H),7.68–7.59(m,2H),7.57–7.41(m,2H),6.74–6. 47(m,3H),6.40(d,J=13.6Hz,1H),4.22(dd,J=16.8,8.2Hz,4H),,2.58–2.53(m,2H),2.28– 2.18(m,2H),1.92(s,12H),1.87–1.70(m,6H),1.66–1.52(m,2H),1.51–1.38(m,2H).MS(ESI + )m / z 731.69[M+H] + (calculated for[C 45 H 51 N2O5S] + :731.35).

[0056] Example 2 Synthesis of Compound 1-2

[0057]

[0058] Compound 1-2 was synthesized using the method described in the literature (J. Med. Chem. 2023, 66, 18, 13332–13345). 1 HNMR (400MHz, DMSO-d6) δ8.57 (t, J = 5.5 Hz, 2H), 8.10 (s, 1H), 8.07 (d, J = 5. 8Hz,2H),7.51–7.40(m,2H),6.64(d,J=8.2Hz,2H),6.27–6.22(m,4H),5.29 (s,2H),4.04–3.89(m,4H),3.27–3.17(m,4H),2.22–2.13(m,4H),1.86–1.7 7(m,2H),1.66–1.58(m,4H),1.55–1.48(m,6H),1.37–1.35(m,54H).MS(ESI + )m / z 1196.44[M+H] + (calculated for[C 62 H 98 N7O 16 ] + :1196.71).

[0059] Example 3 Synthesis of Compound 1-3

[0060]

[0061] Compounds 1-1 (37 mg, 0.050 mmol) and 1-2 (50 mg, 0.042 mmol) were dissolved in iPrOH, and then DMTMM (28 mg, 0.10 mmol) was added and stirred at 40 °C overnight. After the reaction was completed, the solvent was removed under reduced pressure to obtain a crude product. After column chromatography purification (MeOH / CH2Cl2, 1 / 99–15 / 85, v / v), compound 1-3 was obtained as a green solid (71 mg, 89.0%). MS (ESI + )m / z 1910.06[M+H] + (calculated for[C 107 H 146 N9O 20 S] + :1910.04).

[0062] Example 4 Synthesis of Compound 1-4 (ICG-PSMA-D5)

[0063]

[0064] Compound 1-3 (65 mg, 0.034 mmol) was dissolved in a mixed solution of CH2Cl2 (8 mL) and TFA (2 mL) and stirred at room temperature overnight. After the reaction was completed, the solvent was removed under reduced pressure, and the crude product was washed five times with Et2O to obtain 1-4 (ICG-PSMA-D5) as a green solid (44 mg, 82.3%). 1H NMR(600MHz,DMSO-d6)δ12.44(s,6H),10.02(s,1H),8.71(t,J=5.7Hz,2H),8 .29–8.22(m,3H),8.20(d,J=1.5Hz,2H),8.11–8.01(m,4H),7.96(dd,J=20.4, 10.0Hz,2H),7.77(d,J=8.9Hz,2H),7.73–7.66(m,5H),7.64(q,J=8.0Hz,2H) ,7.53–7.45(m,2H),6.57(s,2H),6.50(d,J=13.7Hz,1H),6.41(d,J=13.6Hz,1 H),6.35(dd,J=10.8,8.2Hz,4H),4.28–4.16(m,4H),4.11–4.04(m,4H),3.29 –3.25(m,4H),2.54–2.52(m,2H),2.35(t,J=7.3Hz,2H),2.29–2.21(m,4H),1. 95–1.91(m,2H),1.90(s,6H),1.89(s,6H),1.85–1.76(m,6H),1.72–1.66(m, 6H),1.55(q,J=8.0,7.3Hz,6H),1.51–1.45(m,2H),1.38–1.32(m,4H).MS(ESI + )m / z 1572.68[M+H] + (calculated for[C 83 H 98 N9O 20 S] + :1572.66).

[0065] Example 5 Synthesis of Compound 2-1

[0066]

[0067] Hydrazine benzenesulfonic acid (9.1 g, 60 mmol) and isopropyl methyl ketone (7.4 g, 85 mmol) were heated to reflux in 100 mL of glacial acetic acid for 4 hours. After the reaction was completed, the mixture was cooled to room temperature, the pink solid precipitate was filtered, the precipitate was dissolved in methanol, and potassium hydroxide isopropanol solution was added until the yellow solid was completely precipitated. The yellow solid was filtered, washed with ether 3 times, and dried to obtain an off-white powder, which was the intermediate compound. The product was used directly in the subsequent reaction without purification.

[0068] The above compound and 1,4-butanesulfonate were dissolved in 40 mL of toluene and heated under reflux for 50 h. After the reaction, an off-white solid was obtained by filtration, which was the intermediate compound. The product was used directly in the subsequent reaction without purification.

[0069] Furthermore, the above compound (376mg, 1mmol) and (86 mg, 0.5 mmol) was dissolved in acetic anhydride solution, potassium acetate was added, and the mixture was stirred overnight at 60°C. The product was separated and purified by preparative liquid phase separation to obtain a metallic green solid compound (189 mg, 21.3%). 1 H NMR (400MHz, DMSO-d6) δ8.21(d,J=14.0Hz,2H),7.74(d,J=1.5Hz,2H),7.61(dd,J=8.2,1.5Hz,2H),7.37( d,J=8.3Hz,2H),6.34(d,J=14.2Hz,2H),4.16(t,J=7.3Hz,4H),2.70(t,J=5.9Hz,4H),1.92–1.64(m,12H).

[0070] Example 6 Synthesis of Compound 2-2

[0071]

[0072] Compound 1-2 (120 mg, 0.10 mmol) and p-hydroxyphenylpropionic acid (17 mg, 0.10 mmol) were dissolved in iPrOH, and then DMTMM (55 mg, 0.20 mmol) was added and stirred at 40 °C overnight. After the reaction was completed, the solvent was removed under reduced pressure to obtain a crude product. After column chromatography purification (MeOH / CH2Cl2, 1 / 99–15 / 85, v / v), the tert-butyl protected intermediate compound was obtained as an off-white foamy solid (87 mg, 64.7%). MS (ESI + )m / z 1008.29[M+H] + (calculated for [C 71 H 106 N7O 18 ] + :1344.76).

[0073] Further, the above compound was dissolved in a mixed solution of CH2Cl2 (8 mL) and TFA (4 mL) and stirred at room temperature overnight. After the reaction was completed, the solvent was removed under reduced pressure, and the crude product was washed five times with Et2O to obtain 2-2 as a white solid (52 mg, 86.7%). MS (ESI+ )m / z 1008.29[M+H] + (calculated for[C 47 H 58 N7O 18 ] + :1008.38).

[0074] Example 7 Synthesis of Compound 2-3 (IR783-PSMA-D5)

[0075]

[0076] Compound 2-2 (30 mg, 0.030 mmol) was dissolved in Na2CO3 solution, and compound 2-1 (27 mg, 0.030 mmol) was added thereto, and heated at 75°C for 4 hours. After the reaction was completed, the solvent was removed under reduced pressure, and the crude product was separated by preparative liquid phase and freeze-dried by freeze dryer to obtain 2-3 (IR783-PSMA-D5) as a green solid (23 mg, 41.3%). 1 HNMR(600MHz,DMSO-d6)δ9.94(s,1H),8.77(s,2H),8.21(s,1H),8.13(s,2H),7.79(d,J=13.9Hz,2H),7. 69–7.59(m,6H),7.56(d,J=8.1Hz,2H),7.26(dd,J=20.5,8.0Hz,4H),7.09–7.03(m,2H),6.44–6.31(m,4H ),6.20(d,J=14.2Hz,2H),4.07(s,8H),3.30(s,4H),2.87(s,2H),2.71(s,4H),2.63–2.58(m,2H),2.30–2 .22(m,4H),1.95–1.87(m,4H),1.69(s,12H),1.58(s,8H),1.36(t,J=7.4Hz,4H),1.21(s,14H).HRMS(ESI - )m / z 618.5149[M-3H] 3- (calculated for[C 85 H 102 N9O 30 S4] 3- :618.8544).

[0077] Example 8 Optical Property Measurement

[0078] Fluorescence spectroscopy: Prepare the test substance solution (10 μM) in PBS and use a fluorescence spectrometer to test the fluorescence spectrum and excitation wavelength (λEM ) and emission (λ EX )wavelength.

[0079] The results are shown in Table 1.

[0080] Example 9 Affinity determination with PSMA

[0081] 1) Dilute the test compound to different concentrations using HEPES buffer solution (50 mM):

[0082] 400μM, 40μM, 4μM, 400nM, 40nM, 4nM, 0.4nM, 0.04nM.

[0083] 2) Add 25 μL of the compound to be tested in gradient concentrations, 25 μL of NAAG (160 μM) and 50 μL of PSMA recombinant protein solution (0.4 μg / mL) to the black 96 ELISA plate. After mixing evenly in the ELISA instrument, transfer to a shaker and incubate at 37°C for 1 hour. After the incubation, add 100 μL of OPA detection reagent to each well, incubate in the dark for 3 minutes, and measure the fluorescence intensity at Ex / Em=350 / 450nm.

[0084] 3) The experimentally measured IC is converted to 50 Value converted to K i .

[0085] The activity data are shown in Table 1.

[0086] Table 1 Optical and affinity data of fluorescent probes.

[0087]

[0088] a Affinity is expressed as K i ,mean±SD,n=3.

[0089] Example 10 Cell Imaging Experiment

[0090] 22Rv1 and PC-3 cells were seeded in 12-well plates (22Rv1, 6×10 5 / hole; PC-3,5×10 4 / well) and cultured for 36 hours at 37°C and 5% CO2. Before the experiment, the old culture medium was removed and washed twice with fresh culture medium, and then fluorescent dye (final concentration, 1 μM) solution was added and incubated for 1 hour. After the incubation, the cells were washed twice with PBS buffer, and the cell fluorescence staining images were collected using a fluorescence microscope, using the CY7 filter set.

[0091] The results are as follows Figure 2 .

[0092] Example 11 Near-infrared in vivo fluorescence imaging and in vitro fluorescence imaging of tumor-bearing mice

[0093] Each 22Rv1 tumor-bearing mouse was injected with ICG-PSMA-D5 PBS solution (1 nmol / mouse, 100 μL) through the tail vein, and fluorescence images were collected at different time points (1, 2, 4, 6, 12, 24, 36 and 48 hours) using a near-infrared zone 1 small animal in vivo imager. Filter: λ EX =780nm,λ EM = 845nm. After the in vivo imaging, the main organs and tissues were collected for ex vivo imaging, and the data were collected, processed and analyzed.

[0094] For the inhibition group, DCFPyL (2.26 μmol / mouse) and ICG-PSMA-D5 (1 nmol / mouse) were co-injected into tumor-bearing mice via the tail vein, and fluorescence imaging data were collected under the same experimental conditions for comparison.

[0095] Results Figure 3 The results showed that after ICG-PSMA-D5 was injected into the tail vein, the fluorescent signal accumulated rapidly in the tumor site, which could effectively distinguish the tumor boundary, and the retention time in the tumor tissue was significantly longer than that in the normal tissue. Figure 4 As shown in Figure c, after completing in vivo near-infrared living fluorescence imaging (48 hours), we dissected out the main organs and tissues for ex vivo fluorescence imaging. The signals in tumors and kidneys were significantly higher than those in other organs, further proving that ICG-PSMA-D5 has excellent selectivity and affinity. Tumor ( Figure 4 g) H&E staining results of paraffin sections confirmed poorly differentiated carcinoma. Figure 4 b and 4d) demonstrate that ICG-PSMA-D5 is highly specific for PSMA in vivo.

[0096] Example 12 Near-infrared second-zone in vivo and in vitro fluorescence imaging of tumor-bearing mice

[0097] Each 22Rv1 tumor-bearing mouse was injected with ICG-PSMA-D5 PBS solution (1 nmol / mouse, 100 μL) through the tail vein, and fluorescence images were collected at different time points (1, 2, 4, 6, 8, 12, 24, 30, 36 and 48 hours) using the near-infrared zone II small animal in vivo fluorescence imaging system. The laser wavelength was selected as 808 nm, the filter was selected as 1000 nm, the exposure time was 50 ms, and the ROI was circled using Image J software to calculate and analyze the fluorescence signal values ​​of the main organs.

[0098] For the inhibition group, DCFPyL (2.26 μmol / mouse) and ICG-PSMA-D5 (1 nmol / mouse) were co-injected into tumor-bearing mice via the tail vein, and fluorescence imaging data were collected under the same experimental conditions for comparison.

[0099] Results Figure 4 . The results showed that under the same dosage and collection time, ICG-PSMA-D5 exhibited similar metabolic pathways, but there were also certain differences between the images obtained in the near-infrared zone 1 and zone 2, which may be mainly related to the penetration ability of the probe at different wavelengths and the strength of the fluorescence signal of the probe under different filters. As time goes on, the clearance rate of the probe in normal tissue is faster than that in tumors, and the TNR value continues to increase. At 24 hours after injection and subsequent time points, the TNR value is greater than 3.5, and the tumor edge can be clearly distinguished.

[0100] Example 13 Near-infrared second-zone fluorescence intraoperative navigation experiment

[0101] After completing the near-infrared zone II in vivo fluorescence imaging (48 hours after ICG-PSMA-D5 injection), a mouse was selected and a simulated surgical resection of the tumor of the 22Rv1 tumor-bearing mouse was performed under the guidance of real-time near-infrared zone II fluorescence images. During the entire operation, the mouse was anesthetized, and the near-infrared zone II small animal in vivo fluorescence imaging system was used to record the entire tumor resection process.

[0102] Results Figure 5 A. The results showed that under the real-time guidance of near-infrared zone II fluorescence, the tumor margin can be accurately determined, enabling precise tumor resection during surgery.

[0103] Example 14 Intraoperative Navigation Experiment under Endoscopic System

[0104] 24 hours after the tail vein injection of ICG-PSMA-D5 PBS solution (1nmol / mouse, 100μL), the tumors of LNCaP tumor-bearing mice were simulated surgically removed under the guidance of real-time images of the endoscopic fluorescence imaging system. During the entire operation, the endoscopic fluorescence imaging system was used to record the entire tumor removal process. After the operation, the main organs and tissues were dissected, and the fluorescence signals of the ex vivo organs were collected, and the data were processed and analyzed using Image J software.

[0105] Results Figure 5 B. The results showed that ICG-PSMA-D5 can clearly show the tumor in the surgical field, especially after the skin near the tumor is removed, the edge of the tumor can be accurately judged. Under the guidance of fluorescence, the tumor can be well visualized. After the first large tumor is removed ( Figure 5c) Since the diameter of the second residual part is about 1 mm, it is impossible to distinguish whether the residual part is tumor tissue under white light, but the signal is strong under fluorescence ( Figure 5 d and 5h), it was judged to be a tumor and secondary resection was performed. After the resection was completed, there was no obvious fluorescent signal in the surgical area ( Figure 5 e and 5i). After the surgery, the main organs and tissues were removed for ex vivo fluorescence imaging ( Figure 5 j), the second resected suspicious tissue is smaller in size, so the fluorescence signal is slightly lower than the first resected tumor. Since the operation time is selected 24 hours after drug administration, the signal value in the bladder is low, which can effectively avoid the contamination of the surgical field by urine during the operation.

[0106] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A EuK dimer compound or a pharmaceutically acceptable salt thereof, characterized in that: The structure of the EuK dimer compound is shown in formula (I): Wherein, F is the fluorophore; W is a PSMA targeting ligand or its analogue; L1 is absent or represented by n is an integer from 1 to 12; L2 does not exist or is represented by n is an integer from 1 to 12.

2. The EuK dimer compound or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: F is selected from the following: Wherein, R1 and R2 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R3 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ; n is an integer between 1 and 5; Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R5 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ; n=an integer between 1 and 5; Wherein, R1 and R2 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R3 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ; R4 represents Cl or Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R5 is CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ; R6=Cl or Wherein, R1 and R2 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R3 and R4 are simultaneously or independently CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + ; Wherein, R1, R2, R3 and R4 are simultaneously or independently represented by SO3 - M + or SO3H,M + Mainly selected from Na + , K + , Li + ; R5 and R6 are simultaneously or independently CH3, CH2CH3, CH2CH2CH2SO3H, CH2(CH2)2CH2SO3H, CH2CH2CH2SO3 - M + 、CH2(CH2)2CH2SO3 - M + 、M + Mainly selected from Na + , K + , Li + .

3. The compound according to claim 1 and 2, characterized in that The compound is selected from the group consisting of:

4. The derivative of the EuK dimer compound according to claim 3, characterized in that The derivatives include pharmaceutically acceptable salts of the compounds represented by formula (II) or formula (III).

5. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to any one of claims 1 to 4, and optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

6. A kit, characterized in that: The kit comprises the compound according to any one of claims 1 to 4 or the composition according to claim 5 as an active ingredient.

7. A reagent for diagnosing or detecting abnormal prostate cancer specific membrane antigen (PSMA), characterized in that: The agent comprises the compound according to any one of claims 1 to 4 or the composition according to claim 5 as an active ingredient.

8. A prostate cancer specific membrane antigen (PSMA) targeted inhibitor, characterized in that: The active ingredient is the compound according to any one of claims 1 to 4 or the composition according to claim 5.

9. Use of the reagent according to claims 6 to 8 for diagnosis and / or treatment selected from the group consisting of: preparation of PSMA detection reagents, preparation of prostate cancer detection reagents, diagnosis of prostate cancer, fluorescence navigation in surgical treatment, and detection of clinical samples of prostate cancer.

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