Dimeric compounds targeting psma, derivatives and uses thereof

Abstract

The present application relates to the field of biological medicine, and provide a kind of targeting PSMA dimer compound and its derivative and application, the structure general formula of the dimer compound is as shown in formula (I).The dimer compound and its derivative of the present application have very high affinity and targeting to PSMA, after being labeled with suitable radionuclide, can be used as nuclear medicine molecular probe, to realize early diagnosis, staging and treatment of prostate cancer, 68 Ga]diPSMA-1-DOTA showed good tumor uptake in PET imaging of tumor mice.Therefore, this kind of radioactive complex has good clinical application prospect in tumor imaging targeting PSMA (especially PET imaging), and lays an important foundation for the radiotherapy application of [ 177 Lu]diPSMA-1-DOTA and [ 225 Ac]diPSMA-1-DOTA.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, and in particular to a dimer compound targeting PSMA, derivatives and applications thereof. BACKGROUND

[0002] Prostate cancer is one of the most common cancers affecting men and the fifth leading cause of male cancer death. According to reports, there were more than 1.1 million new cases and 300,000 deaths worldwide in 2020. This disease is more common in older men, with a median age of over 60 years at diagnosis. Prostate cancer is a major medical problem that needs attention because the disease has a long incubation period and has a high incidence and mortality rate. Therefore, research on early diagnosis, initial staging, biochemical recurrence detection and treatment of prostate cancer is of great significance. Thanks to diagnostic tests such as PSA testing and tissue biopsy and early treatment, the mortality rate of the disease has been steadily declining.

[0003] In order to accurately diagnose prostate cancer, a series of 11 C and 18 F-labeled choline and acetate derivatives have been synthesized, but they need to be improved in terms of accuracy, specificity and sensitivity in diagnosis. For example, 18 F] fluoroacetylcholine positron emission tomography / computed tomography (PET / CT) has a detection sensitivity of only 71% for recurrent prostate cancer. Therefore, there is a need to further develop new nuclear medicine molecular probes with higher specificity for prostate cancer.

[0004] Prostate-specific membrane antigen (PSMA) is a transmembrane glycoprotein enzyme that is selectively highly expressed in prostate cancer cells, also known as type I folate hydrolase or type II glutamate carboxypeptidase. Studies have shown that the expression level of PSMA in prostate cancer is 100-1000 times higher than that in healthy prostate tissue, and is highly positively correlated with the degree of cancer progression, making it an important target for the diagnosis and treatment of prostate cancer.

[0005] Currently, many PSMA small molecule inhibitors with lysine-urea-glutamate (EuK) as the pharmacophore have entered clinical research and application. This inhibitor was first reported in 2001 and was first introduced by Professor Pomper's laboratory at Johns Hopkins University School of Medicine in the United States in 2002 for research in the field of nuclear medicine diagnosis and treatment of prostate cancer. The main probes related to the current are [ 68 Ga]-PSMA-11, [ 68 Ga]-PSMA-617, [ 18 F]-PSMA-1007, [ 18F]-DCFPyL, Al 18 F-PSMA-BCH, etc., wherein, 68 Ga]-PSMA-11 and 18 F]-DCFPyL has been approved by the US FDA, and interferes with the diagnostic accuracy of part of the lesion. These probes provide a powerful tool for precise staging of prostate cancer, precise positioning of biochemical recurrence lesions, and treatment of prostate cancer, but both have high bladder uptake.

[0006] Further studies have found that PSMA dimer compounds show good application potential. For example, compared with [ 68 Ga]-PSMA-11, its dimer shows longer retention time, stronger PSMA affinity and higher tumor / background ratio (EJNMMI Res 2, 23, 2012); compared with [ 68 Ga]-PSMA-11 and 18 F]-PSMA-1007, 18 F-Bi-PSMA has better pharmacokinetics, higher tumor uptake and better imaging quality (European Journal of Medicinal Chemistry, 221, 113502, 2021); dimer 68 Ga-DOTA-DiPSMA has good pharmacokinetics and high imaging ability, and has low non-specific uptake in normal organs such as salivary glands and kidneys, showing potential application in radioligand therapy (Frontiers in Bioengineering and Biotechnology, 9, 811972, 2022).

[0007] However, there are few dimer compounds with clinical application prospects at present, and there is no dimer probe that has been truly applied in clinical. From the perspective of 68 Ga-DOTA-DiPSMA imaging results, it has obvious imaging in large blood vessels; moreover, the maximum absorption of the probe in the lesion of prostate cancer patients is only SUV max =4.41. Therefore, the development of PSMA probes with excellent pharmacokinetic properties will provide efficient tools for the detection of prostate cancer lesions and the subsequent application of radiotherapy, and has broad clinical prospects and important use value. SUMMARY

[0008] In view of the deficiencies in the prior art, the present application provides a dimer compound targeting PSMA and its derivatives and applications.

[0009] In a first aspect, the present application provides a dimeric compound targeting PSMA or a pharmaceutically acceptable derivative thereof, the general structure of which is shown as Formula (I):

[0010]

[0011] wherein R is DOTA or NODA.

[0012] Specifically, when R is DOTA, the structure of the dimeric compound is shown as Formula (II), denoted as diPSMA-1-DOTA.

[0013]

[0014] When R is NODA, the structure of the dimeric compound is shown as Formula (III), denoted as diPSMA-1-NODA.

[0015]

[0016] The dimeric compound of the present application or a pharmaceutically acceptable derivative thereof can also be referred to as a ligand compound or a labeling precursor.

[0017] The present application introduces a biphenyl structure in the molecule, which increases the liposolubility of the molecule and further improves the biophysical and chemical properties of the pharmacophore. In addition, using DOTA or NODA as a chelating group of radionuclide, the prepared radioimaging reagent has high affinity and targeting to PSMA, and has good tumor uptake in tumor mice and prostate cancer patients. Therefore, after labeling different nuclides, this kind of molecule has good clinical application prospect in the field of tumor imaging and radiotherapy of PSMA targeting.

[0018] It should be noted that the dimeric compound of the present application can exist in multiple crystal forms, i.e. different lattice arrangements of the same element composition of the compound. Polymorphs usually have different X-ray diffraction spectra, infrared spectra, melting points, densities, hardness, crystal form, optical and electrical properties, stability and solubility. Different factors such as recrystallization solvent, crystallization rate and storage temperature can obtain recrystallization products dominated by single crystal form. It can be understood that the dimeric compound of the present application includes all such crystal forms.

[0019] The dimer compounds of the present application can have chiral centers and / or axial chirality, and thus exist in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds and single diastereomers, and the form of cis-trans isomers. Each chiral center or axial chirality will independently produce two optically active isomers, and all possible optically active isomers and diastereomeric mixtures, and pure or partially pure compounds are included in the scope of the present application. The dimer compounds of the present application include all the above-mentioned isomeric forms thereof.

[0020] The pharmaceutically acceptable derivatives of the present application include forms such as salts, solvates, esters / ethers, prodrugs and metabolites, etc.

[0021] The "salts" and "esters / ethers" each refer to the form in which a compound exists, which does not cause significant irritation to the organism to which it is administered, and does not cause the biological activity and properties of the compound to disappear.

[0022] The "solvate" refers to a compound containing a stoichiometric or non-stoichiometric amount of solvent, and is selectively formed in the crystallization process with a pharmaceutically acceptable solvent such as water or other solvents such as ethanol, etc.

[0023] The "prodrug" is also known as precursor drug, drug precursor, precursor drug, etc., which refers to a compound obtained by chemical structural modification of a drug, which is inactive or less active in vitro, and releases active drugs in vivo through enzymatic or non-enzymatic conversion to exert drug efficacy. There are two major categories of prodrugs: one is carrier prodrug, simply referred to as carrier prodrug; the other is biological prodrug. The carrier prodrug refers to a compound with activity and its transport carrier combined by covalent bond, which is released by simple hydrolysis in vivo to exert pharmacological effect by active compounds. The carrier prodrug is often less active or inactive compared to the parent compound. For the structure of the carrier, it is lipophilic, requires no harm to the organism, and can release active compounds in time. The biological prodrug is different from the carrier prodrug, and the active substance does not need to be temporarily combined with the carrier, but to exert the effect through the change of its own molecular structure. The biological prodrug itself has no activity, and the active metabolite is in the organism.

[0024] In a second aspect, the present application provides a preparation method of the above-mentioned dimer compound.

[0025] The synthesis route of diPSMA-1-DOTA is shown in Figure 1 The synthesis route of diPSMA-1-NODA is shown in Figure 2 The specific synthesis steps are described in detail in the specific embodiments section.

[0026] It can be understood that, based on the preparation of the dimeric compound, the person skilled in the art can prepare the pharmaceutically acceptable derivatives of the dimeric compound according to the prior art and the experimental skills mastered by the person skilled in the art.

[0027] In a third aspect, the present application provides use of the dimeric compound or the pharmaceutically acceptable derivative thereof in the preparation of a medicament for diagnosing and / or treating one or more tumors or cells expressing PSMA.

[0028] Further, the medicament is a chemical medicament, a nucleic acid medicament or a protein medicament.

[0029] It should be noted that the above-mentioned medicament should be understood as a broad-sense medicament, and any chemical substance capable of affecting the physiological functions of organs and the metabolic activities of cells belongs to the category of medicaments, including substances used for preventing, treating and diagnosing diseases.

[0030] Further, when the dimeric compound or the pharmaceutically acceptable derivative thereof is modified with a diagnostic and / or therapeutic group, the formed substance can be used as a corresponding diagnostic and / or therapeutic reagent and / or medicament. In addition, the present application does not have a special limitation on the specific form of the diagnosis and treatment, which is completely dependent on the modified group.

[0031] In some embodiments of the present application, the form of diagnosis includes nuclide imaging; further, the nuclide imaging can be PET imaging or SPECT imaging. PET imaging is Positron emission tomography (PET), and SPECT imaging is single-photon emission computerized tomography (SPECT).

[0032] In some embodiments of the present application, the form of treatment includes radiotherapy.

[0033] In a fourth aspect, the present application provides a PSMA-targeting inhibitor, which includes the dimeric compound or the pharmaceutically acceptable derivative thereof.

[0034] In the embodiments of the present application, the PSMA-targeting inhibitor can include only one or more of the dimeric compound or the pharmaceutically acceptable derivative thereof, or can include other known PSMA-targeting inhibitors in addition to the dimeric compound or the pharmaceutically acceptable derivative thereof.

[0035] In a fifth aspect, the present application provides a nuclear medicine molecular imaging probe targeting PSMA, which includes a nuclide-labeled dimeric compound or a pharmaceutically acceptable derivative thereof.

[0036] In a preferred embodiment of the present application, the nuclear medicine molecular imaging probe is [ 68 Ga]diPSMA-1-DOTA, having the structure shown in formula (IV):

[0037]

[0038] In another preferred embodiment of the present application, the nuclear medicine molecular imaging probe is [ 18 F]diPSMA-1-NODA, having the structure shown in formula (V):

[0039]

[0040] In a sixth aspect, the present application provides a PSMA-targeting radionuclide therapeutic drug, comprising the above-mentioned dimer compound or a pharmaceutically acceptable derivative thereof labeled with a radionuclide.

[0041] In an embodiment of the present application, the radionuclide therapeutic drug is [ 177 Lu]diPSMA-1-DOTA, having the structure shown in formula (VI):

[0042]

[0043] In another embodiment of the present application, the radionuclide therapeutic drug is [ 225 Ac]diPSMA-1-DOTA, having the structure shown in formula (VII):

[0044]

[0045] The present application provides a PSMA-targeting dimer compound and its derivatives and applications, which have high affinity and targeting property to PSMA. After being labeled with a suitable radionuclide, the dimer compound and its derivatives can be used as a nuclear medicine molecular probe to achieve early diagnosis, staging and treatment of prostate cancer. These radioactive complexes have high stability in physiological saline and mouse serum. More importantly, these radioactive complexes are simple and fast to prepare, have high labeling rate, and have [ 68 Ga]diPSMA-1-DOTA shows good tumor uptake in PET imaging of tumor mice. Therefore, these radioactive complexes have good clinical application prospects in tumor imaging targeting PSMA (especially PET imaging), which also lays an important foundation for the radiotherapy application of [ 177 Lu]diPSMA-1-DOTA and [ 225 Ac]diPSMA-1-DOTA. BRIEF DESCRIPTION OF DRAWINGS

[0046] Figure 1 The synthetic route for diPSMA-1-DOTA is as follows, with the following reagents and conditions: (a) 1) K2CO3, Pd(pph3)4, N2, THF / H2O; 2) NaOH, phosphoric acid; (b) H2, Pd / C, MeOH; (c) Na2CO3, H2O / THF; (d) 1) HATU, DIPEA, DMF; 2) 5% piperidine, DMF; (e) 5% piperidine, DMF; (f) HATU, DIPEA, DMF; (g) HATU, DIPEA, DMF; (h) TFA, H2O;

[0047] Figure 2 The synthetic route for diPSMA-1-NODA is shown, with the following reagents and conditions: (a) HATU, DIPEA, DMF; (b) TFA, H2O.

[0048] Figure 3 Mass spectrometry results for diPSMA-1-DOTA;

[0049] Figure 4 LC-MS detection image of diPSMA-1-DOTA;

[0050] Figure 5 Mass spectrometry results for diPSMA-1-NODA;

[0051] Figure 6 LC-MS detection image of diPSMA-1-NODA;

[0052] Figure 7 The results show the affinity determination of diPSMA-1-DOTA, diPSMA-1-NODA, and PMSA-617.

[0053] Figure 8 for[ 68 PET images of Ga]diPSMA-1-DOTA in mice with LNCap and PC3 tumors at 60 minutes;

[0054] Figure 9 for[ 68 Ga]diPSMA-1-DOTA in prostate cancer patients and 68 Comparison of head-to-head imaging using Ga-PSMA-11. Detailed Implementation

[0055] Unless otherwise defined or unless required by particular context, all technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0056] The words "comprise," "comprising," "include," "including," and "contains," "containing," and the like are to be construed in an open, inclusive way, meaning "including but not limited to."

[0057] The expressions "one embodiment," "an embodiment,” or the like as used herein do not necessarily refer to the same embodiment, though they can. Neither are such expressions necessarily referring to particular embodiments of any inventively disclosed subject matter. The expressions can refer to at least one implementation of the particular feature or characteristic, or a particular combination of features or characteristics, as variously described in connection with the corresponding expressions. These expressions are also to be construed to cover all existing forms or variants of the particular features, features, or characteristics, including those that are developed after the calendar dates of the embodiments described.

[0058] In order to make the objects, technical solutions and advantages of the present application clearer, the technical solutions in the present application will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present application, and not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by a person of ordinary skill in the art without creative effort belong to the protection scope of the present application.

[0059] Unless otherwise specified in the embodiments, the techniques or conditions are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions. Unless otherwise specified, the reagents or instruments are conventional products that can be purchased through a regular channel.

[0060] In the following examples, some abbreviations correspond to the following Chinese full names:

[0061] Pd(pph3)4: tetrakis(triphenylphosphine)palladium;

[0062] THF: tetrahydrofuran;

[0063] DCM: dichloromethane;

[0064] MeOH: methanol;

[0065] Fmoc-osu: 9-fluorenylmethyl succinimidyl carbonate;

[0066] DMF: N,N-dimethylformamide;

[0067] HATU: 2-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate;

[0068] DIPEA: N,N-diisopropylethylamine;

[0069] TFA: trifluoroacetic acid;

[0070] OPA: o-phthalaldehyde;

[0071] NAAG: N-acetyl aspartate glutamate.

[0072] Example 1: Preparation of diPSMA-1-DOTA (synthetic route as shown in Figure 1

[0073]

[0074] Synthesis of compound 1: 250ml single-necked flask was added 1-bromo-4- nitrobenzene (1g, 1equ), 3,5-bis(methoxycarbonyl)phenylboronic acid pinacol ester (1.9g, 1.2equ), K2CO3(1.03g, 1.5equ) and Pd(pph3)4(0.286g, 0.05equ), after adding nitrogen protection, THF (45ml) and water (15ml) were added, after adding, the temperature was raised to 80°C and reacted for 3h, LC-MS detection showed that the reaction was completed. The reaction solution was concentrated, and after drying, 200ml DCM and 100ml water were added, the organic phase was concentrated and dried, then THF 50ml, water 50ml and 2M sodium hydroxide aqueous solution 100ml were added, after stirring for 3h, the layers were separated, the aqueous phase was washed with DCM twice, the aqueous phase was adjusted to pH=3 with 5% phosphoric acid, a large amount of solid was precipitated, which was filtered, the solid was washed with water twice and then dried under vacuum, finally 10.7g of compound was obtained, the yield was 50%, MS m / z: [M+H] + calcd for C14H10NO6, 288.04, found 288.5.

[0075]

[0076] Synthesis of compound 2: 250ml single-necked flask was added compound 1 (500mg, 1equ), Pd / C 10% (50mg, 0.1equ), 20ml MeOH, after adding, hydrogen was replaced for 3 times, and the reaction was carried out at room temperature overnight, LC-MS detection showed that the reaction was completed. Filtration and concentration of the filtrate gave the product 250mg, yield 55%, MS m / z: [M] calcd for C14H11NO4, 256.07, found 257.24.

[0077]

[0078] ​Synthesis of compound 3: 100 ml single neck flask was charged with compound 2 (250 mg, 1 equ), H20 / THF (20 ml) and Na2C03(124 mg, 1.2 equ), after addition, Fmoc-osu (361 mg, 1.1 equ) was added, after addition, the reaction was carried out at room temperature for 3 hours, and LC-MS was used to detect the end of the reaction. The dry reaction liquid was concentrated, 50 ml DCM and 30 ml water were added, and the aqueous phase was extracted with 50 ml DCM once more. The combined organic phase was concentrated and dried over silica gel column, and the eluent polarity was DCM (500 ml), DCM / MeOH = 200:1 (500 ml), 100:1 (200 ml) to 50:1 (100 ml) in order. The product was obtained in 300 mg, the yield was 64%, MS m / z: [M+H] = 479.3, calcd for C29H21NO6, 479.14. + calcd for C29H21NO6, 479.14, found 479.3.

[0079]

[0080] Synthesis of compound 5: 100 ml single neck flask was charged with compound 3 (230 mg, 1 equ), tri-tBu-EUK (468 mg, 2 equ), 10 ml DMF, DIPEA (344 ul, 4 equ), and finally HATU (402 mg, 2.2 equ) was added, after addition, the reaction was carried out at room temperature for 1 hour, and LC-MS was used to detect the end of the reaction to obtain compound 4. Subsequently, an equal volume of 5% piperidine and DMF mixture was added to the reaction liquid, dried and washed with ether 3 times, and then dried and purified by reverse phase to obtain the product compound 5 450 mg, the yield was 78%, MS m / z: [M+H] = 1195.9, calcd for C62H97N7O16, 1195.70. + calcd for C62H97N7O16, 1195.70, found 1195.9.

[0081]

[0082] Synthesis of compound 6: 100 ml single neck flask was charged with compound 5 (450 mg, 1 equ), Fmoc-5-amino pentanoic acid (Fmoc-osu, 129 mg, 1 equ), 10 ml DMF, DIPEA (272 ul, 4 equ), and finally HATU (159 mg, 1.1 equ) was added, after addition, the reaction was carried out at room temperature for 1 hour, and LC-MS was used to detect the end of the reaction. An equal volume of 5% piperidine and DMF mixture was added to the reaction liquid, dried and washed with ether 3 times, and then dried and purified by reverse phase to obtain the product 340 mg, the yield was 69.8%, MS m / z: [M+H] = 1295.60, calcd for C67H107N8O17, 1295.77. + calcd for C67H107N8O17, 1295.77, found 1295.60.

[0083]

[0084] Synthesis of compound diPSMA-1-DOTA: 100ml single neck flask was charged with compound 6 (170mg, 1equ), tri-tBu-DOTA (75mg, 1equ), 10ml DMF, DIPEA (93ul, 4equ), lastly HATU (55mg, 1.1equ), after addition, reaction was carried out at room temperature for 1h, LC-MS was used to monitor the reaction, compound 7 was obtained. The reaction solution was concentrated to dryness, 10ml TFA, 0.5ml H2O was added, stirred for 3h, ether was added to precipitate the solid, centrifuged, washed with ether twice, dried, and the product was obtained by reverse phase purification, 100mg, yield 56.8%. The mass spectrum is shown in Figure 3 , LC-MS detection is shown in Figure 4 . MS m / z: [M+2H] + / 2calcd for (C59H86N12O24) / 2, 673.29, found 673.4.

[0085] Example 2: Preparation of diPSMA-1-NODA

[0086]

[0087] Compound diPSMA-1-NODA was prepared according to the synthetic route shown in Figure 2 , 97mg, yield 61%. The mass spectrum is shown in Figure 5 , LC-MS detection is shown in Figure 6 . MS m / z: [M+2H] + / 2calcd for (C55H79N11O22) / 2, 622.76, found 622.8.

[0088] Example 3: Preparation of [ 68 Ga]diPSMA-1-DOTA and stability test

[0089] [ 68 Ga]diPSMA-1-DOTA was labeled using the miniallinone module, the brief method is as follows: after obtaining the 68 Ga eluent from the Ge-Ga generator, it was added to the acetate / sodium acetate buffer (pH = 7.4) containing 50μg of precursor diPSMA-1-DOTA, heated at 100℃ for 10min, the crude product was enriched by C18, washed with water, diluted with ethanol, and filtered through a sterile filter to obtain

[0090] The stability test method is as follows: the obtained probe solution is respectively placed in physiological saline and mouse serum and incubated at 37 degrees, and its radioactivity purity is analyzed by HPLC at 2 hours and 4 hours, respectively, and the results show that the radioactivity ligand[ 68 Ga]diPSMA-1-DOTA has good in vitro stability, and after 4 hours, its radioactivity purity in physiological saline and mouse serum is >99%.

[0091] Example 4: 18 Preparation of [ 18 F]diPSMA-1-NODA

[0092] Physiological saline is eluted to obtain 18 F, and then added to a potassium hydrogen phthalate solution and an AlCl3 solution, shaken and shaken uniformly, placed for 5 min, then the diPSMA-1-NODA precursor is added, heated at 110°C for 15 min. The crude product is enriched by C18, washed with water, and then eluted and diluted with ethanol, filtered with a sterile filter membrane to obtain the final product.

[0093] Example 5: 177 Preparation of [ 225 Lu]diPSMA-1-DOTA and [ 177 Ac]diPSMA-1-DOTA

[0094] Preparation of [ 177 Lu]diPSMA-1-DOTA and [ 225 Ac]diPSMA-1-DOTA is similar to the preparation of [ 68 Ga]diPSMA-1-DOTA, 177 Lu or 225 Ac is added to a buffer solution containing 50 μg of the precursor dissolved in 100°C, heated for 10 min, and then the crude product is enriched by C18, washed with water, and then eluted and diluted with ethanol, filtered with a sterile filter membrane to obtain the final product.

[0095] Example 6: Activity determination

[0096] The activity of the ligand compound is determined by the "OPA-Ki detection" experiment, and the specific steps are as follows:

[0097] a. Preparation of solution:

[0098] (1) Borax buffer: 4.7625 g of sodium tetraborate (MW: 381) is dissolved in 200 mL of deionized water, the pH is adjusted to 10.0 with NaOH, and finally the volume is made to 250 mL with a volumetric flask, and stored at 4 degrees for use.

[0099] (2) OPA detection reagent: 10 mg OPA was dissolved in 100 mL of methanol, diluted to 10 mL with the solution in (1), and finally added with 25 mL of 2-mercaptoethanol. After mixing, it was stored at 4 degrees in the dark.

[0100] (3) HEPES buffer: 50 mM HEPES; 0.1 M NaCl; pH 7.5.

[0101] b. Ki determination:

[0102] (First open the incubator and set the temperature to 37 degrees. While waiting for the temperature to rise, you can prepare the following solutions)

[0103] (1) PSMA recombinant protein dilution: dilute the protein to a concentration of 0.4 mg / mL with HEPES buffer for standby;

[0104] (2) NAAG dilution: dilute NAAG to 160 mM with HEPES buffer for standby;

[0105] (3) Probe dilution: dilute the probe to different concentrations: 40 mM, 4 mM, 400 nM, 40 nM, 4 nM, 400 pM, 4 pM with HEPES buffer;

[0106] (4) Add 12.5 mL of NAAG, 12.5 mL of probe and 25 mL of protein in a 96-well plate and mix well. Incubate the mixed solution at 37 degrees for 1 hour. Set 3 parallel groups for each group.

[0107] (5) Add 50 mL of prepared OPA detection reagent to each tube, then incubate at room temperature for 3 minutes in the dark.

[0108] (6) Detection conditions: Enzyme marker, Ex / Em = 350 / 450 nm, gain 100.

[0109] As shown in Figure 7 , the ligand compound has high affinity for PSMA, and the Ki values of diPSMA-1-DOTA and diPSMA-1-NODA are 0.27 nM and 0.76 nM, respectively, which are higher than the activity of PSMA-617 compound measured under the same conditions. Therefore, the activity of the dimer compound is obviously improved compared with the monomer compound, which lays a foundation for further PET imaging application.

[0110] Example 7: 68 PET imaging of [68Ga]diPSMA-1-DOTA

[0111] Take the mice implanted with LNCaP and PC3 tumors, respectively, and inject 0.1 mL of radioactive complex [68 Ga]diPSMA-1-DOTA solution (approximately 15 MBq) was injected, and micro-PET imaging was performed 60 minutes after injection. Anesthesia was administered using 2.5% isoflurane before and during imaging, and the imaging time was 15 minutes.

[0112] like Figure 8 As shown, significant radioactivity concentration was observed in LNCaP-tumor mice, with a tumor / tumor ratio reaching 37. At 60 minutes, the radiocomplex also showed significant concentrations in the kidneys and bladder, while concentrations were lower in other organs.

[0113] Will 68 Ga-DOTA-DiPSMA was used in PET imaging of the model mice using the same method, and the tumor / tumor ratio was significantly lower than that of […]. 68 Ga]diPSMA-1-DOTA, and, from 68 The imaging results of Ga-DOTA-DiPSMA show that it has a clear presence in large blood vessels.

[0114] Example 8: [ 68 Clinical PET imaging of Ga]diPSMA-1-DOTA

[0115] Radioactive complex was administered intravenously. 68 Ga]diPSMA-1-DOTA solution was administered to a 65-year-old male prostate cancer patient (diagnosed with prostate cancer by SerumPSA). Clinical PET imaging was performed 30 minutes after injection, with a imaging time of 20 minutes. One day later, a head-to-head comparison was performed. 68 Ga-PSMA-11 imaging.

[0116] like Figure 9 As shown, [ 68 Ga]diPSMA-1-DOTA uptake in tumors is SUV max =15.69, in imaging of the same patient, commercially approved probes 68 The value of Ga-PSMA-11 is 13.76.

[0117] The probe of this invention was used to perform identical imaging comparisons in multiple cases willing to undergo clinical trials. 68 Ga-diPSMA-1-DOTA uptake in tumors is SUV max =15.69-35.21, in imaging of the same patient, commercially approved probes 68 The Ga-PSMA-11 values ​​range from 13.76 to 33.97, all of which are values ​​found in the probe SUV of this invention. max The value is larger. Additionally, 68The maximum absorption of the Ga-DOTA-DiPSMA probe in the lesion of the prostate cancer patient is SUV max = 4.41.

[0118] Therefore, it can be seen from the clinical data that the probe of the present application has 68 Ga]diPSMA-1-DOTA has excellent clinical effect, and through safety experiment evaluation, the safety of the probe of the present application is higher, that is, it has good application prospect. 68 Ga]diPSMA-1-DOTA has excellent clinical effect, and through safety experiment evaluation, the safety of the probe of the present application is higher, that is, it has good application prospect.

[0119] Finally, it should be pointed out that: the above examples are only used to illustrate the technical solutions of the present application, and not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A dimeric compound targeting PSMA, characterized in that, The dimer compounds are diPSMA-1-DOTA and diPSMA-1-NODA, and the structural formulas of the diPSMA-1-DOTA and diPSMA-1-NODA are as follows: diPSMA-1-DOTA; diPSMA-1-NODA.

2. Use of a radioactive complex formed after radionuclide labeling of the dimer compound of claim 1, the radioactive complex having a structure as shown in formula (IV): in the preparation of a medicament for diagnosing one or more tumors or cells expressing PSMA: Formula (IV) The diagnostic form is nuclide imaging.

3. Use according to claim 2, characterized in that, The medicament is a chemical drug, a nucleic acid drug, or a protein drug.

4. Use according to claim 2, characterized in that, The nuclide imaging is positron emission tomography.

5. A nuclear medicine molecular imaging probe targeting PSMA, characterized in that, The nuclear medicine molecular imaging probe has a structure as shown in formula (IV): Formula (IV).

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