CN114014843B - PSMA targeted nuclide/fluorescent bimodal ligand, molecular probe and application - Google Patents

PSMA targeted nuclide/fluorescent bimodal ligand, molecular probe and application Download PDF

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CN114014843B
CN114014843B CN202111363417.0A CN202111363417A CN114014843B CN 114014843 B CN114014843 B CN 114014843B CN 202111363417 A CN202111363417 A CN 202111363417A CN 114014843 B CN114014843 B CN 114014843B
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psma
nuclide
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杨兴
田捷
张宁
胡振华
段小江
李源
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Peking University First Hospital
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Abstract

The invention belongs to the field of nuclear medicine, and relates to a PSMA targeted nuclide/fluorescent bimodal ligand, a molecular probe and application. The PSMA targeted nuclide/fluorescent bimodal ligand has a structure shown in a formula I, and R is a nuclide chelating group; q is a fluorophore; n is 0 to 6. The invention takes dencichine-ureido as a target structure of PSMA, designs and synthesizes a series of bimodal molecular probes targeting PSMA by optimizing a linker and then performing nuclide/fluorescence double functionalization, and performs detailed evaluation in vitro and in vivo. The nuclide/fluorescence bimodal probe can be used for preoperative PET/CT imaging to formulate an operation route, perform fluorescence guidance during operation, perform accurate tumor positioning and boundary division, and improve postoperative curative effect.

Description

PSMA (patterned beam-selective membrane antigen) targeted nuclide/fluorescent bimodal ligand, molecular probe and application
Technical Field
The invention belongs to the field of nuclear medicine, and particularly relates to a PSMA (patterned beam splitter) targeted nuclide/fluorescent bimodal ligand, a PSMA targeted nuclide/fluorescent bimodal molecular probe and application thereof.
Background
Prostate cancer (PCa) is taken as the second largest cancer of men all over the world, the total number of patients is close to 130 million every year, the number of the patients accounts for 13.5 percent of new cancers, and with the extension of the life span of people in China, the westernization of dietary structure and living habits, the incidence rate of prostate cancer in China is in a rapid rising trend, and the prostate cancer becomes one of the problems threatening the health of men. Accurate detection of early stages of prostate cancer is critical to the therapeutic efficacy and survival of patients.
Prostate cancer is relatively slow in progress and is often difficult to find early, Prostate Specific Antigen (PSA) combined magnetic resonance imaging and needle biopsy can effectively diagnose PCa, but currently, a functional imaging device with high sensitivity and specificity capable of defining biological behaviors of the prostate cancer is lacked, PET/CT has been developed greatly in the last decade, PET/CT integrates a nuclear medicine molecular image with high sensitivity and specificity and a CT fine anatomical image, and great potential has been brought forward in multiple aspects such as preoperative diagnosis and staging of primary prostate cancer and treatment decision making. In terms of treatment, radical prostatectomy is one of the most effective methods for treating localized and locally advanced prostate cancer, and the operator determines the margin range mainly by means of preoperative imaging examination, intraoperative naked eye observation and exploration and the experience of the operator. The normal tissues can be damaged due to the overlarge surgical excision range, and the normal functions such as urine control and the like are influenced; however, if the excision range is too small, a positive margin is caused, and the patient is easy to relapse. Therefore, how to preserve as much normal tissue and function as possible during surgery while performing a thorough resection of the prostate cancer affected area is a problem that clinicians often need to face and address.
The navigation technology in the targeted fluorescence is a good solution to the problem, and in the field of prostate cancer, a Prostate Specific Membrane Antigen (PSMA), which is a prostate cancer molecular marker with high specificity, exists. PSMA is overexpressed in 90% of prostate cancers, while normal tissues such as the lacrimal salivary glands, renal proximal tubules are expressed only in small amounts. The expression level of PSMA in prostate cancer is highly correlated with tumor invasion and malignancy. Therefore, PSMA becomes an ideal biomarker for accurate localization imaging and intraoperative navigation of prostate cancer lesions. The PSMA-targeted near-infrared fluorescence surgical navigation technology mainly utilizes the advantages that near-infrared dyes have longer wavelength, stronger tissue penetrating power, smaller scattering and the like, can light the focus in the operation, and enables operators to clearly know the range of the focus so as to more completely and effectively excise the focus, so that the PSMA-targeted near-infrared fluorescence surgical navigation technology is a very promising surgical mode for treating prostate cancer. Furthermore, the optical-nuclear medicine bimodal molecular imaging probe can combine the advantages of two imaging modalities, and has high sensitivity and high accuracy, lesion tissues can be identified and positioned through the nuclear medicine modality before operation, and the position of a focus is accurately marked by fluorescence in the operation to guide the excision of the surgical operation.
The current nuclide diagnosis and treatment and operation navigation medicines targeting PSMA used in clinical experiments are all glutamic acid-carbamido as basic skeletons. These probes have good effect on PSMAThe targeting property and affinity of the cancer cell are improved, but the problems of serious retention of bladder nuclein and high background of nuclein in kidney exist, the early diagnosis accuracy rate of the prostate cancer is seriously influenced by the retention of a large amount of nuclein in the bladder, and the background of double-kidney high nuclein has certain toxic and side effects on the kidney. Therefore, the development of a high-specificity low-renal-excretion PSMA molecular imaging probe by changing the structure of a medicament is urgently needed, and the probe has important significance for realizing accurate positioning and grading of prostate cancer focuses. The subject group developed a new generation of ODAP-ureido PSMA inhibitors, which increased the binding of the PSMA molecular imaging probe to 80pM, which had lower renal excretion properties. The novel ODAP-ureido is used as a core structure, and the in vivo metabolic kinetics characteristic of the drug is optimized by changing linker to adjust lipophilicity on the premise of keeping high affinity with PSMA. The subject group develops a probe with ODAP-Urea-Lys as a brand new targeting structure 68 Ga-P137, with classical glutamic acid probes 68 Compared with Ga-PSMA617, the probe obviously reduces the retention of radioactivity in bladder while keeping the uptake in tumor area, and is beneficial to the accurate diagnosis of prostate cancer in situ.
Disclosure of Invention
The invention aims to provide a novel PSMA targeted nuclide/fluorescent bimodal ligand, a molecular probe and application.
In order to achieve the above object, the present invention provides a PSMA targeted nuclide/fluorescent bimodal ligand, which has a structure shown in formula I,
Figure BDA0003359713380000031
wherein,
r is a nuclide chelating group;
q is a fluorophore;
n is 0 to 6, specifically 0, 1, 2, 3, 4, 5, 6, preferably 0, 1, 2, 3, 4, 5, more preferably 2, 3, 4.
According to the present invention, the nuclide chelating group generally refers to a group formed by a bifunctional chelating agent, which may be DOTA, NOTA, NODA, nodaa, DOTP, TETA, ATSM, PTSM, EDTA, EC, HBEDCC, DTPA, BAPEN, Df, DFO, TACN, NO2A, NOTAM, CB-DO2A, Cyclen, DO3A, DO3AP, HYNIC, MAS3, MAG3 or isonitrile.
The structures of the above bifunctional chelating agents are well known to those skilled in the art, for example, the DOTA and NOTA structures are shown below, respectively:
Figure BDA0003359713380000032
according to some preferred embodiments of the present invention, the fluorescent group is a near-infrared fluorescent group, and the near-infrared dye has advantages of longer wavelength, stronger tissue penetration, less scattering and the like, so that the fluorescent group can be suitable for surgical navigation. Preferably, the near-infrared fluorescent group is a fluorescent group in a near-infrared first region (NIR-I, 700-1000nm) and/or a near-infrared second region (NIR-II, 1000-1700 nm).
According to a preferred embodiment of the present invention, the fluorophore is represented as follows, and the fluorophore has both near-infrared first-zone fluorescence and near-infrared second-zone fluorescence, and is better suitable for surgical navigation:
Figure BDA0003359713380000041
more specifically, the PSMA-targeting bimodal ligand preferably has the structure shown in formula II, wherein n is 0, 1, 2, 3, 4, 5, 6, preferably 0, 1, 2, 3, 4, 5, more preferably 2, 3, 4.
Figure BDA0003359713380000042
The bimodal ligands of the present invention can be prepared using the synthetic route shown in FIG. 2-1. The reaction conditions are as follows: (a) triphosgene, triethylamine, anhydrous DCM; (b) triethylamine, anhydrous DCM; (c) Pd/C, H 2 Methanol; (d) NaHCO 2 3 Dioxane, water; (e) n, N-diisopropylethylamine, 2-chlorotrityl resin, DCM, DMF; (f) a solution of 20% piperidine in DMF and,DMF solution of Fmoc-3- (2-naphthyl) -L-alanine, HBTU, HOBt and DIPEA; (g) 20% piperidine in DMF, Fmoc- (4-aminomethyl) benzoic acid, HBTU, HOBt and DIPEA in DMF; (h) 20% piperidine in DMF, DDE-Fmoc-L-lysine, HBTU, HOBt and DIPEA in DMF; (i) 20% piperidine in DMF, DOTA, HBTU, HOBt and DIPEA in DMF; (j) 2% Hydrazine (Hydrazine) in DMF, Fmoc-L-aspartic acid 1-tert-butyl ester, HBTU, HOBt and DIPEA in DMF; (k) 20% piperidine in DMF, LY-12, PyBOP and DIPEA in DMF; (l) Trifluoroacetic acid, water and triisopropylsilane. The starting materials used are either commercially available or are prepared by conventional organic synthesis methods.
The second aspect of the present invention provides a PSMA-targeted nuclide/fluorescent bimodal molecular probe, which is the above PSMA-targeted nuclide/fluorescent bimodal ligand labeled with a radionuclide, specifically, the radionuclide is chelated with the nuclide chelating group.
The bimodal molecular probe can be prepared by labeling a PSMA targeting bimodal ligand with a radionuclide, specifically, the ligand is dissolved in a radioactive labeling buffer solution, and then different radionuclides are added for reaction to obtain the corresponding molecular probe.
According to the invention, the radionuclide may be a diagnostic radionuclide or a therapeutic radionuclide.
According to some preferred embodiments of the invention, the diagnostic radionuclide is 68 Ga、 64 Cu、 18 F、 86 Y、 90 Y、 89 Zr、 111 In、 99m Tc、 11 C、 123 I、 125 I and 124 at least one of I.
According to some preferred embodiments of the invention, the therapeutic radionuclide is 177 Lu、 125 I、 131 I、 211 At、 111 In、 153 Sm、 186 Re、 188 Re、 67 Cu、 212 Pb、 225 Ac、 213 Bi、 212 Bi and 212 at least one of Pb.
The invention also provides application of the PSMA targeted nuclide/fluorescent bimodal ligand or the PSMA targeted nuclide/fluorescent bimodal molecular probe in preparation of a nuclide diagnosis and treatment reagent and/or a surgical navigation drug for targeting PSMA.
The invention takes dencichine-ureido as a target structure of PSMA, designs and synthesizes a series of bimodal molecular probes targeting PSMA by optimizing a linker and then performing nuclide/fluorescence double functionalization, and performs detailed evaluation in vitro and in vivo. The nuclide/fluorescence bimodal probe can be used for preoperative PET/CT imaging to formulate an operation route, perform fluorescence guidance during operation, perform accurate tumor positioning and boundary division, and improve postoperative curative effect, thereby being applied to clinic.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Drawings
The above and other objects, features and advantages of the present invention will become more apparent by describing in more detail exemplary embodiments thereof with reference to the attached drawings.
FIG. 1 shows the structural formulae of LY-13 to LY-16; LY-13, n ═ 0; LY-14, n ═ 2; LY-15, n is 4; LY-16, n is 6.
FIG. 2-1 shows a general route for preparation of LY-13 to LY-16.
FIG. 2-2 shows a route for the preparation of LY-12.
FIGS. 3-1 to 3-5 are a mass spectrum of LY-12, a mass spectrum of LY-13, a mass spectrum of LY-14, a mass spectrum of LY-15, and a mass spectrum of LY-16, respectively.
FIG. 4 shows the Ki values of LY-13-16.
Fig. 5 shows the results of the LNCaP and PC3 cell experiments.
FIG. 6 shows 68 Results of Ga-labeled LY-13-16 tumor-bearing mouse PET/CT.
FIG. 7 shows 68 Results of biodistribution 60min after Ga-LY-15 injection and 120min after injection, and inhibition experiment result, data is tissue% ID/g + -SD, n is 4.
FIG. 8 shows the fluorescence image of LY-15 bearing mice.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below. While the following describes preferred embodiments of the present invention, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein.
Synthesis of ODAP-PSMA targeting bimodal ligand
Preparation of ligands LY-13, LY-14, LY-15, LY-16
The structures of the four ODAP-PSMA ligands (LY-13 to LY-16) are shown in FIG. 1, the general synthetic route is shown in FIG. 2-1, the reagents are purchased from reagents company, and are not purified, and the coupling of amino acids is performed according to the standard Fmoc solid phase synthesis method.
Reaction conditions are as follows: (a) triphosgene, triethylamine, anhydrous DCM; (b) triethylamine, anhydrous DCM; (c) Pd/C, H 2 Methanol; (d) NaHCO 2 3 Dioxane, water; (e) n, N-diisopropylethylamine, 2-chlorotrityl resin, DCM, DMF; (f) 20% piperidine in DMF, Fmoc-3- (2-naphthyl) -L-alanine, HBTU, HOBt and DIPEA in DMF; (g) 20% piperidine in DMF, Fmoc- (4-aminomethyl) benzoic acid, HBTU, HOBt and DIPEA in DMF; (h) 20% piperidine in DMF, DDE-Fmoc-L-lysine, HBTU, HOBt and DIPEA in DMF; (i) 20% piperidine in DMF, DOTA, HBTU, HOBt and DIPEA in DMF; (j) 2% Hydrazine (Hydrazine) in DMF, Fmoc-L-aspartic acid 1-tert-butyl ester, HBTU, HOBt and DIPEA in DMF; (k) 20% piperidine in DMF, LY-12, PyBOP and DIPEA in DMF; (l) Trifluoroacetic acid, water and triisopropylsilane.
Preparation of ligands LY-13 to LY-16: as shown in FIG. 2-1, 100mg of the resin (0.03mmol) was taken out of a 10mL solid phase synthesis tube, and 2mL of Dichloromethane (DCM) was added for swelling and repeated three times for 5 minutes, followed by washing three times for 5 minutes with N, N-Dimethylformamide (DMF). The amino protecting group Fmoc was removed using 20% piperidine in DMF (v/v) in 2mL 20% piperidine in DMF for 2 min, 10min, followed by 3-5 washes with 2mL DMF for 2 min each. The Fmoc amino acid of 3 times chemical amount was activated with HBTU or PyBOP of 3.6 times chemical amount in the presence of DIPEA of 7.2 times chemical amount with respect to the resin (0.03mmol), and then added to the synthesis tube, and reacted for 1 hour with electromagnetic stirring. All deprotection, activation and coupling steps are carried out as described above, except for the step of deprotecting the Dde. The amino protecting group Dde was removed using 2% hydrazine in DMF (v/v) by 2mL of 2% hydrazine in DMF for 2 min, 3 min, and 3 min. The cleavage of the ligand from the resin and the removal of the tert-butyl ester were accomplished by stirring for 2 hours using 5mL of trifluoroacetic acid/triisopropylsilane/water (95:2.5:2.5, v/v/v) and washing the resin with 2mL of trifluoroacetic acid, collecting all filtrates, removing the trifluoroacetic acid under reduced pressure, and preparing the crude product by reverse phase HPLC, followed by lyophilization to give the desired ligands LY-13 to LY-16. The ligand structures were identified by mass spectrometry and are shown in FIGS. 3-2 through 3-5 and Table 1, respectively.
Preparation of resin LY-12
The preparation route is shown in figure 2-2. Reaction conditions are as follows: (a) potassium carbonate, acetonitrile, 50 ℃; (b) trifluoroacetic acid, dichloromethane.
The preparation process comprises the following steps: neoindocyanine green (17, 200mg, 0.24mmol) and tert-butyl p-hydroxybenzoate (18, 45mg, 0.26mmol) were taken in a 50mL round-bottomed flask, 20mL of acetonitrile was added to dissolve, potassium carbonate (50mg,0.36mmol) was added, the mixture was heated to 50 ℃ under electromagnetic stirring, and the reaction was stopped after 20 hours. Preparation by reverse phase HPLC gave a green solid (LY-18, 134mg, 0.13 mmol). The product obtained in the previous step is taken out of a round-bottom flask, 4mL of dichloromethane and 4mL of trifluoroacetic acid are added, and the reaction is carried out for 3 hours under the electromagnetic stirring. After completion of the reaction, the solvent was removed to give a green solid (LY-12, 120mg, yield 100%). The structure was identified by mass spectrometry as shown in FIG. 3-1 and Table 1.
Table 1 summarizes the mass spectra results for compounds LY-12, 13, 14, 15, 16 as follows:
Figure BDA0003359713380000081
preparation of resin LY-6
N6-Cbz-L-lysine benzyl ester hydrochloride (1, 1g, 2.46mmol) and triethylamine (995mg, 9.85mmol) were dissolved in 50mL DCM, the mixture was added dropwise slowly to a solution of triphosgene (240mg, 0.82mmol) in DCM under an ice salt bath, after completion of the dropwise addition, reaction was continued at room temperature for 3h, and Compound 2(708mg, 2.46mmol, prepared according to this laboratory patent CN 109748896B and patent CN 111233758B) and triethylamine (745mg, 7.38mmol) were added to the reaction solution, which was purified by column to give LY-3(1.23g) as a colorless oil. LY-3 was debenzylated with hydrogen in methanol under catalysis of 100mg 10% Pd/C to give LY-4 as a colorless oil (744mg) after purification of the crude product on a silica gel column. LY-4(300mg, 0.65mmol) was dissolved in 20mL dioxane/water (2.5/1, v/v), sodium bicarbonate (164mg, 1.95mmol) and Fmoc-Cl (201mg, 0.78mmol) were added, and the mixture was stirred at room temperature for 15 minutes. After the reaction, 200mL of ethyl acetate was added to the reaction solution, washed twice with water, the organic phase was collected, dried over anhydrous sodium sulfate and the solvent was removed under reduced pressure, and the crude product was purified by a silica gel column to obtain a white solid product LY-5(290 mg). 1g of 2-CTC resin was taken in a 50mL solid phase synthesis tube, swelled with dichloromethane for 1 hour, after solvent was drained, LY-5(290mg) in DCM/DMF (1:1, v/v) was added, after 3 hours reaction at room temperature, blocked four times with DCM/MeOH/DIPEA (10:10:1, v/v/v) for 10 minutes each, washed with methanol and dried to constant weight, yielding 1.2 g of resin LY-6, with a loading of 0.33 mmol/g. Reference is made to the present laboratory patent CN 109748896B for the preparation of resin LY-6.
Ki determination of LY-13 to LY-16
1. Preparation of the solution
(1) Borax buffer solution
4.7625g of sodium tetraborate is dissolved in 200mL of deionized water, the pH value is adjusted to 10.0 by NaOH, and finally the volume is adjusted to 250mL by a volumetric flask and the sodium tetraborate is stored at 4 ℃ for standby.
(2) OPA detection reagent
10mg of OPA was dissolved in 100. mu.L of methanol, diluted to 10mL with the solution obtained in step (1), and 25. mu.L of 2-mercaptoethanol was added thereto, mixed well and stored at 4 ℃ in the dark.
(3)HEPES buffer
50mM HEPES；0.1M NaCl；pH 7.5。
2. Measurement procedure
The PSMA recombinant protein was diluted to 0.4. mu.g/mL with HEPES buffer, NAAG was diluted to 160. mu.M with HEPES buffer, and the probe was diluted to various concentrations with HEPES buffer (400. mu.M, 40. mu.M, 4. mu.M, 400nM, 40nM, 4nM, 400pM, 40 pM). Add 25. mu.L NAAG, 25. mu.L probe and 50. mu.L protein to EP tube, centrifuge for 3-5 seconds, mix the solution at 37 ℃ and incubate for 1 hour, each set of 3 replicates. After the incubation, the reaction was terminated by denaturing the protein at high temperature by metal bath at 95 ℃ for 5 minutes. Then 100. mu.L of the prepared OPA detection reagent is added into each tube, vortexed for 2-3 seconds to be fully and uniformly mixed, and incubated for 3 minutes at room temperature in a dark place. mu.L of the solution was taken out of each tube, added to a 96-well blackboard, and immediately detected by a microplate reader (detection conditions: Ex/Em. 350/450nm, gain 100).
Binding affinities of LY-13 to LY-16 to PSMA as shown in table 2 and fig. 4, Ki values for all compounds were nanomolar, showing high affinity to PSMA with potential for further applications.
TABLE 2 Ki values for LY-13 to LY-16
Complexes Ki(nM)
LY-13 5.28nM
LY-14 1.54nM
LY-15 1.78nM
LY-16 3.09nM
Marking and quality control
Marking:
68 ga: 1.0mg of ligand was accurately weighed into a sample tube, dissolved by adding 200. mu.L of DMSO (dimethyl sulfoxide), and the ligand concentration was diluted to 5. mu.g/. mu.L. Pipette 6. mu.L of ligand solution and 130. mu.L of NaOAc solution (1mol/L) into a vial, add 2mL of freshly eluted solution 68 Ga 3+ An ionic solution (hydrochloric acid solution with a solvent of 0.05mol/L and a radioactivity of 10-17mCi) was shaken up, sealed and reacted at 90 ℃ for 10 minutes. The reaction solution was cooled to room temperature and analyzed by HPLC for quality control.
Quality control:
68 the radiochemical purity of the Ga complexes was determined by HPLC (high performance liquid chromatography) with a mobile phase of 40% acetonitrile in water (containing 0.1% TFA), all complexes having a radiochemical purity of greater than 90% and being able to be studied next without purification.
68 Cell binding and internalization experiments for Ga-labeled products
LNCaP cells and PC3 cells were seeded separately in 24-well plates, each plate being seeded at 1X 10 5 Placing the cells in a container containing 5% CO 2 And then the cells become monolayer adherent cells in a constant temperature incubator at 37 ℃. The radioactive complex was diluted to 7.4MBq/mL using the medium, and 50. mu.L each (containing 0.37MBq radioactivity) was taken in three small tubes as total activity. After the old medium was aspirated, the cells were washed twice with fresh medium, 400 μ L of fresh medium was added to each well, 50 μ L of fresh medium was added to the experimental group (n ═ 6), 50 μ L of 1mmol/L inhibitor (ZJ-43) was added to the inhibitory group (n ═ 6), and incubation was carried out at 37 ℃ for 15 min; after incubation 0.37MBq per well was added 68 Ga-labeled complex (50. mu.L) was incubated at 37 ℃ for 1 h; after incubation, the medium was aspirated, washed 2 times with 1mL of cold PBS containing 0.2% BSA, then the experimental group was washed 2 times with 50mM glycine NaCl solution at pH 2.8 for 5min, glycine solutions were collected, and finally the experimental group and the inhibitory group were incubated with 0.5M NaOH solution for 10min to disrupt the cells, NaOH solutions were collected, and counted separately with a gamma counter.
The results of the cell experiments are shown in figure 5, 68 the uptake of Ga-LY-13-16 in PSMA positive cells (LNCaP) was significantly higher than that of PSMA negative cells (PC3) and could be inhibited by ZJ-43, suggesting that 68 Ga-LY-13-16 binds specifically to PSMA. 68 Ga-LY-13-16 has a high internalization ratio in LNCaP cells, between 63.1% and 81.9%.
68 Imaging of Ga-labelled products
Taking 0.1mL of the newly prepared 68 Ga-labeled complex (5.6-7.4 MBq) is injected into Balb/c nude mice with male 22RV1 tumor (the tumor diameter is about 0.6 cm) through tail vein, and is anesthetized with isoflurane after 1h and 2h respectively, and then the imaging of small animal PET/CT (SUPER-NOVA, Pingsheng technology, China) is carried out, and the SUV of standard uptake value is sketched for the region of interest.
68 The PET-CT imaging and data of Ga-LY-13-16 in tumor-bearing mice are shown in figure 6 and table 3, and the results show that: 68 the Ga complexes can be obviously concentrated in the tumor area, wherein 68 Ga-LY-15 and 68 the uptake value of Ga-LY-14 in the tumor area is higher than that in the tumor area 68 Ga-LY-13、 68 Ga-LY-16, in particular, 68 the ratio of Ga-LY-15 tumor/muscle and tumor/liver is higher than that of Ga-LY-15 tumor/muscle 68 Ga-LY-13、 68 Ga-LY-14、 68 Ga-LY-16。
Figure BDA0003359713380000131
68 Biodistribution of Ga-LY-15
Taking 0.1mL of the newly prepared 68 Ga-LY-15(0.74MBq) is injected into Balb/c nude mice with 22RV1 tumor male through tail vein, and in the inhibition experiment, 50mg/kg of ZJ43 (a common PSMA protein inhibitor) is injected through tail vein of the mice half an hour in advance. Injection of drugs 68 After 60min, 120min, mice were sacrificed by decapitation, tumors, muscles and other tissues and organs of interest were removed for measurement by weighing and radioactive counting, and finallyThe percent injected dose per gram (% ID/g) for each tissue was calculated.
68 The biodistribution data of Ga-LY-15 in tumor-bearing mice are shown in Table 4 and FIG. 7, and the results show that: after the injection was carried out for 2 hours, 68 Ga-LY-15 is mainly concentrated in the tumor area and has certain uptake in the kidney of a metabolic organ, 68 the uptake of Ga-LY-15 in the tumor was 8.98 + -1.91% ID/g, and the tumor to muscle ratio was 7.47 + -1.42. In an inhibition experiment, ZJ43 can obviously inhibit tumor pairs 68 Uptake of Ga-LY-15 (8.98% ID/g vs 2.13% ID/g), indicating 68 The specific binding of Ga-LY-15 to PSMA protein is shown by the results 68 Ga-LY-15 is a very potential radionuclide imaging agent for prostate cancer.
TABLE 4 68 Biodistribution of Ga-LY-15 tumor-bearing mice
Figure BDA0003359713380000141
Figure BDA0003359713380000151
Fluorescent imaging of LY-15
The fluorescence imaging data of LY-15 in tumor-bearing mice are shown in FIG. 8 and Table 5, and the results show that: the probe LY-15 can be quickly targeted to a tumor region, mainly gathers in the tumor region and urinary system organs including kidney and bladder, and simultaneously the probe in blood circulation can be quickly eliminated so as to reduce background signals, after the injection of LY-15 intravenously, the fluorescence signal of the tumor region is strongest at 1h, the fluorescence signal is obviously weakened after 4h, and the tumor/muscle display is optimal at 8 h.
Table 5LY-15 ROI values in tumor, muscle and their ratios (mean ± SD, n ═ 4)
Figure BDA0003359713380000152
Having described embodiments of the present invention, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.

Claims (5)

1. A PSMA targeted nuclide/fluorescent bimodal ligand has a structure shown in a formula I,
Figure FDA0003737874190000011
wherein,
r is a nuclide chelating group;
q is a fluorophore;
n is 0 to 6;
the nuclide chelating group is a group formed by a bifunctional chelating agent, and the bifunctional chelating agent is DOTA, NOTA, NODA, NODAGA, DOTP, TETA, NOTAM or DO3 AP;
the structure of the fluorophore is shown as follows:
Figure FDA0003737874190000012
2. the PSMA-targeting nuclide/fluorescent bimodal ligand of claim 1, wherein n is 0, 1, 2, 3, 4, 5.
3. The PSMA targeted nuclide/fluorescent bimodal ligand as in any of claims 1-2, wherein the PSMA targeted bimodal ligand has a structure represented by formula II,
Figure FDA0003737874190000021
4.a PSMA-targeted nuclide/fluorescent bimodal molecular probe that is a radionuclide-labeled PSMA-targeted nuclide/fluorescent bimodal ligand of any one of claims 1 to 3; the radionuclide is a diagnostic radionuclide or a therapeutic radionuclide; the diagnostic radionuclide is 68 Ga、 64 Cu、 18 F、 86 Y、 90 Y、 89 Zr、 111 In、 99m Tc、 11 C、 123 I、 125 I and 124 at least one of I; the therapeutic radionuclide is 177 Lu、 125 I、 131 I、 211 At、 111 In、 153 Sm、 186 Re、 188 Re、 67 Cu、 212 Pb、 225 Ac、 213 Bi、 212 Bi and 212 at least one of Pb.
5. Use of a PSMA-targeting nuclide/fluorescent bimodal ligand according to any of claims 1 to 3 or a PSMA-targeting nuclide/fluorescent bimodal molecular probe according to claim 4 for the preparation of a nuclide diagnostic and therapeutic agent and/or a surgical guidance drug targeting PSMA.
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