A labeled precursor targeting prostate-specific membrane antigen and uses thereof

Abstract

This invention discloses a labeled prodrug precursor compound for a radiopharmaceutical targeting prostate-specific membrane antigen (PSMA) and its application, belonging to the field of PMA-targeted radiopharmaceutical development technology. This invention utilizes computer-aided drug design technology combined with experiments. While retaining the core region of the "glutamate-urea group," it introduces β-amino acids and employs a linker modification strategy to design a series of PMA-targeting molecules, enriching the variety of PMA-targeting labeled prodrug precursor compounds. Compared with traditional PSMA-617 targeting tracers, the novel PMA-targeting tracer provided by this invention exhibits higher tumor uptake rates, demonstrating greater advantages in clinical diagnosis and treatment. Compared with traditional PSMA-617 targeting tracers, the novel PSMA-targeting tracer provided by this invention also shows no significant increase in radioactive signal uptake values ​​in various non-target organs, resulting in higher safety and greater application value in clinical treatment.

Description

Technical Field

[0001] This invention belongs to the field of prostate-specific membrane antigen inhibitor technology, specifically relating to a labeled prodrug precursor compound that targets prostate-specific membrane antigens and its application. Background Technology

[0002] Prostate cancer is one of the most common tumors of the male genitourinary system. According to the latest global cancer statistics, its incidence rate ranks second among men, only after lung cancer, and it is gradually becoming a major disease threatening men's lives and health. Currently, the comprehensive assessment of prostate cancer patients in clinical practice mainly relies on serum prostate-specific antigen (PSA) level testing, Gleason score, and TNM staging. However, these methods have drawbacks such as high invasiveness, poor timeliness, and insufficient specificity. Most patients are diagnosed at an advanced stage, missing the optimal treatment window, and are prone to multiple metastatic lesions throughout the body, limiting the effectiveness of surgery and chemotherapy. Therefore, there is an urgent need to explore new targeted molecular markers and effective treatment methods for prostate cancer, enabling early detection, early diagnosis, and early treatment to improve prognosis and increase patient benefits.

[0003] Prostate-specific membrane antigen (PSMA), also known as glutamate carboxypeptidase II (GCP II), is a type II transmembrane protein that promotes neurotransmission of glutamate and absorption of folic acid. PSMA is overexpressed on the surface of prostate cancer cells and in their metastases, but its expression is extremely low in non-target organs such as the lacrimal gland, kidney, nervous system, duodenum, and colon. Studies have reported that PSMA expression levels increase with prostate cancer progression and Gleason score elevation. Furthermore, even in metastatic castration-resistant prostate cancer (mCRPC), PSMA expression remains high, while PSA levels are low. Therefore, PSMA is an ideal molecular target for the diagnosis and treatment of prostate cancer, possessing significant scientific research and clinical application value.

[0004] Positron emission tomography (PET) technology has seen rapid clinical development in recent years. It can achieve non-invasive imaging of various diseases and is currently the most advanced molecular imaging technology. It is widely used in the diagnosis, differentiation and treatment evaluation of tumors. 68Ga is a commonly used PET nuclide, which is a positron-emitting radionuclide compared to nuclides that require a cyclotron to obtain. 68 Ga has a suitable half-life (T) 1 / 2 =68min) and can be produced using a germanium-gallium generator, making it easy to obtain. The marking method has advantages such as simplicity, mild conditions, and speed, enabling kit-like production at low cost. 68 Ga-labeled tracers are widely studied and applied globally. In recent years, several domestic institutions have successively introduced [the technology / method / initiatives]. 68 Ge / 68 Ga generator, and in 68 Ga-labeled imaging agents have made encouraging progress in clinical applications. On the other hand, single-photon emission computed tomography (SPECT) technology is relatively mature and can achieve non-invasive imaging of various diseases, which is of great significance for the early diagnosis, staging, prognosis and treatment of diseases. 177 Lu is an ideal therapeutic radionuclide with a half-life of 6.7 days. It emits high-energy beta rays with a short range, which can effectively kill diseased tissues. It can also emit gamma rays for in vivo localization imaging and monitoring of therapeutic effects. It is suitable for SPECT imaging and has a wide range of applications in the targeted radiotherapy of clinical diseases.

[0005] In recent years, thanks to the rapid development of PET and SPECT technologies, an increasing number of radiolabeled tracers targeting prostate-specific membrane antigens (PSMAs) have been widely used in the clinical practice of early diagnosis, preoperative staging, efficacy evaluation, and recurrence and metastasis detection in prostate cancer, achieving significant clinical results. Currently, several PSMA-targeted radiopharmaceuticals have been approved for marketing in Europe and the United States. In terms of diagnosis, [ 18 In a clinical trial involving 139 positive lesions in 8 patients with metastatic prostate cancer, PEC-CT was used to observe [F]DCFPyL. 18 F]DCFPyL showed significant uptake in 138 definite lesions and 1 indistinct lesion, while conventional imaging detected only 45 lesions; in terms of treatment, a clinical trial in Germany involving 99 patients showed that after dosing at 8-12 week intervals... 177Following Lu-PSMA-617, 45% of patients experienced a PSA decrease of over 50%. The diagnostic and therapeutic value of PSMA-targeted radiopharmaceuticals in mCRPC has been fully validated in numerous clinical trials, and the development of PSMA-targeted radiopharmaceuticals has significant clinical value and practical implications. Recent studies have shown that a series of small molecule compounds targeting prostate-specific membrane antigens, constructed based on the glutamate-urea-lysine core unit, exhibit high affinity for PSMA. These compounds, utilizing various radionuclides, can be used for the diagnosis and treatment of prostate cancer. In 2022, Novartis announced that its […]. 177 LuLu-PSMA-617 has been approved by the FDA, providing a powerful imaging tool for the precise imaging localization and treatment of tumor lesions in patients with metastatic and castration-resistant prostate cancer.

[0006] However, at present [ 177 The clinical performance of Lu-PSMA-617 is not entirely satisfactory. Absorption in non-target organs is a major source of side effects. Studies have shown that although PSMA expression levels in healthy tissues (such as salivary glands, kidneys, and small intestines) are significantly lower than in tumor tissues, various healthy tissues such as lacrimal glands, salivary glands, large intestine, kidneys, rectum, and bladder walls can affect […]. 177 Ingestion of Lu-PSMA-617 may lead to toxicity. (The last sentence appears to be incomplete and possibly refers to a separate topic.) 177 During Lu-PSMA-617 treatment, a so-called "tumor pooling effect" was observed in individual cases, where the tumor volume decreased, leading to an increase in the dose absorbed by the kidneys. This has attracted widespread attention. How to reduce the dose absorbed by non-target organs such as the kidneys is a research direction for improving the safety of PSMA and expanding its clinical application.

[0007] To meet the increasing demands of clinical practice in the nuclear medicine diagnosis and treatment of prostate cancer, such as increasing tracer uptake at lesion sites, reducing radioactive concentration in non-target tissues, and improving the sensitivity of detecting small-volume cancer lesions, new small-molecule PSMA inhibitors are constantly being developed and have entered clinical research stages. Therefore, developing a novel radionuclide imaging reagent targeting prostate-specific membrane antigens with high target affinity, tumor uptake rate, and good in vivo metabolism will provide a highly efficient tool for the clinical diagnosis and treatment of prostate cancer, possessing significant clinical translational value and broad application prospects. 177Lu-PSMA-617 is a PSMA-targeted radiopharmaceutical based on a "glutamate-urea" core structure. Its pharmacophore pocket (S1' pocket and zinc active site) interacts effectively with the P1' motif and ZBG, a prerequisite for ensuring the inhibitory activity of the substrate molecule. β-amino acids are important building blocks regulating peptide bioactivity, conformational preference, and physicochemical properties. Compared to α-amino acids, β-amino acids possess anti-peptidase activity, thus increasing serum half-life and exhibiting higher resistance to enzymatic degradation. Furthermore, the introduction of β-amino acids helps regulate the binding affinity between the active molecule and the target protein, thereby enhancing affinity. 1-Naphthyl-L-alanine is a key component of the PSMA-617 structure; removal of the naphthyl group leads to a sharp decrease in tumor uptake of the tracer. et al. evaluated the effects of modifying the hydrophobic linker between the glutamate-urea-lysine binding group and the DOTA chelator, highlighting the importance of 1-naphthyl-L-alanine for pharmacological properties. Subtle changes in 1-naphthyl-L-alanine can lead to significant alterations in the in vivo and in vitro properties of PSMA inhibitors, providing important insights for PSMA inhibitor modification. However, PSMA inhibitors modified with β-naphthyl-alanine have not yet been reported. Among the many clinical application indicators of PSMA inhibitors, affinity is one of the most critical, determining the targeting specificity and thus influencing their application as diagnostic reagents or therapeutic drugs. Researchers have found that, unlike the highly conserved core structure, the functional groups linked to the S1 pocket have a far less significant impact on catalytic activity than the core structure. Therefore, by retaining the targeting motif (P1' region and the zinc-binding group ZBG), introducing β-type modified amino acids of 1-naphthyl-L-alanine can further improve the drug's tumor affinity and uptake value, and reduce uptake in non-target organs. This is of great significance for the development of PSMA-targeted radiopharmaceuticals. Summary of the Invention

[0008] The purpose of this invention is to provide a labeled precursor compound that targets prostate-specific membrane antigens and its application.

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] One of the technical solutions of this invention is to provide a compound that targets prostate-specific membrane antigen and its pharmaceutically acceptable salt, wherein the general structural formula of the compound that targets prostate-specific membrane antigen is shown in Formula I:

[0011]

[0012] In Formula I, x is any integer from 0, 1, 2, 3, 4, or 5; m is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; L1 and L2 are independently selected from straight-chain or branched C1-C4 alkyl groups, -NHCO-, -CH2-O-CH2-, natural amino acids, non-natural amino acids, or one of the following structures, where a is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10:

[0013]

[0014] R is a chelating agent, preferably DOTA, DOTAGA, DOTAM, DO3AP, or DO3AP. PrA DO3AP ABn HYNIC, DiAmSar, NOTA, NOTAGA, TRAPH, NOPO, TRAP-Pr, TRAP-OH, TRAP-Ph, AAZTA, H2DEPDPA, TETA, TE2A, CB-TE2A, 177, 178, 179, PCTA, EDTMP, DTPA, PEPA, HEHA, HBED or HBED-CC TFP, the specific structures are shown below:

[0015]

[0016] Preferably, the compound targeting prostate-specific membrane antigen has any one of the following structural formulas;

[0017]

[0018] More preferably, the structural formula of the compound targeting prostate-specific membrane antigen is selected from:

[0019]

[0020]

[0021] The method for synthesizing the aforementioned compound can be summarized as follows:

[0022]

[0023] First, compound 1 and DIPEA were added to 2-chloro-triphenylmethyl resin to couple it to the resin, yielding compound 2. The resin was washed and dried. 1 mmol of bis(tert-butyl)-L-glutamic acid hydrochloride (compound 3) and 0.5 mL of N-ethyldiisopropylamine (DIPEA) were added to 70 mL of dry CH2Cl2. 0.34 mmol of triphosgene was added to the solution over 4 hours at 0°C to obtain compound 4. The reaction was continued at 25°C with stirring for 1 hour. 0.2 mmol of resin (compound 2) was added to 2 mL of DCM, followed by compound 4. The reaction was carried out at room temperature for 16 hours to obtain compound 5. After the reaction was complete, the resin was filtered out, and 35 mg of tetra(triphenyl)palladium and 200 μL of morpholine were added to 2 mL of... The allyloxycarbonyl protecting group was removed by a solution in CH2Cl2, and the reaction was carried out at room temperature for 3 hours to obtain compound 6. Subunits were then sequentially coupled using the SPPS method, followed by cleavage to obtain a crude PET tracer precursor compound. The crude precursor compound was purified by HPLC to obtain a PET tracer precursor compound targeting prostate-specific membrane antigens.

[0024] Furthermore, the present invention also provides a diagnostic radionuclide marker, which is a radiolabeled compound of the aforementioned compound or an acceptable salt thereof, wherein the radionuclide is a diagnostic or therapeutic radionuclide selected from... 43 Sc、 44 Sc、 51 Mn, 52 Mn, 64 Cu、 67 Ga、 68 Ga、 90 Y、 89 Zr、 99m Tc, 111 ln、 152 Tb, 155 Tb, 201 Tl、 203 Pb, 212 Pb, 213 Bi、 212 Bi、 225 Ac、 211 At、 186 Re、 67 Cu、 188 Re、 177 Lu、 18 F, 76 Br、 77 Br、 123 I, 124 I, 125 I or 11 At least one of C;

[0025] Preferably, the structural formula of the diagnostic radionuclide marker is selected from:

[0026]

[0027]

[0028] More preferably, the structural formula of the diagnostic radionuclide marker is selected from:

[0029]

[0030] In another aspect, the radionuclide labeling method provided by the present invention involves eluting the radionuclide eluent, mixing it with the labeling precursor compound, adjusting the pH to 4-10 using a buffer solution, wherein the buffer solution is sodium acetate, water, ethanol, phosphate, salt buffer solution or dimethyl sulfoxide; reacting at 20-110°C for 10 minutes to overnight;

[0031] In another aspect, the present invention provides the use of a prostate-specific membrane antigen-targeting labeling compound and its diagnostic and therapeutic radionuclide labeling in the preparation of one or more drugs or reagents for the diagnosis and / or treatment of one or more tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammation, metabolic / endocrine disorders or neurological diseases;

[0032] Furthermore, the diagnostic and therapeutic radionuclide-labeled compounds provided by this invention are used to diagnose and treat diseases by the following pathogenic cells: prostate tumor cells, metastatic prostate tumor cells, lung tumor cells, kidney tumor cells, liver tumor cells, glioblastoma cells, pancreatic tumor cells, bladder tumor cells, sarcoma cells, melanoma cells, breast tumor cells, colon tumor cells, germ cells, pheochromocytoma, esophageal tumor cells, and gastric tumor cells;

[0033] Furthermore, the diagnostic method provided by the present invention is radionuclide imaging, which is further preferably PET imaging and SPECT imaging;

[0034] In another aspect, the treatment provided by the present invention is selected from radionuclide therapy, and more preferably includes beta-nuclide therapy and alpha-nuclide therapy;

[0035] The beneficial technical effects of the present invention are as follows:

[0036] This invention combines computer-aided drug design technology with experiments to design a series of prostate-specific membrane antigen-targeting molecules while retaining the core region "glutamate-urea group", thus enriching the types of PSMA-targeted labeling precursor compounds.

[0037] Compared with PSMA-617, the novel prostate-specific membrane antigen targeted tracer provided by this invention has a higher tumor uptake rate and no significant increase in the radioactive signal uptake values ​​of various non-target organs. It has high safety in clinical treatment and is more advantageous in clinical diagnosis and treatment.

[0038] The reagent kit invention involved in this invention includes the following aspects:

[0039] Aspect 1: A radiolabeling kit for prostate-specific membrane antigen-targeting precursor compounds, comprising...

[0040] A certain amount of acetate or its salt buffer is provided to equilibrate the acidic pH eluent from the Ga-68 generator to a pH value in the range of 3 to 5 when the Ga-68 generator / Ltene-177 accelerator is eluted in the kit.

[0041] The chelate functional group of the prostate-specific membrane antigen-targeting precursor compound is capable of chelating gallium-68 / lutetium-177 under radiolabeling conditions; and a metal inhibitor, which is a co-chelating agent capable of inactivating contaminating metals without interfering with the chelation between gallium-68 / lutetium-177 and the PSMA-targeting precursor compound under said radiolabeling conditions, wherein the metal inhibitor is selected from the group consisting of DOTA, DTPA, monosaccharides, and disaccharides, and the PSMA-targeting precursor compound and the chelate are from claims 1-5;

[0042] Aspect 2: The radiolabeled reagent kit for the prostate-specific membrane antigen-targeting precursor compound according to Aspect 1, wherein the acetate, the PSMA-targeting precursor compound, and the metal inhibitor are co-lyophilized;

[0043] Aspect 3: A radiolabeling kit for a prostate-specific membrane antigen-targeting precursor compound according to Aspect 1, wherein the prostate-specific membrane antigen-targeting precursor compound and the metal inhibitor are co-lyophilized, and wherein the acetate or its buffer solution is present alone and may be added subsequently;

[0044] Aspect 4: A radiolabeling kit for prostate-specific membrane antigen-targeting precursor compounds according to any one of Aspects 1 to 3, wherein the acetate or its buffer is present at a specific concentration to equilibrate the acidic eluent from the gallium-68 generator / lutetium-177 accelerator to a pH in the range of 3 to 5 when the generator is eluted in the kit after the addition of hydrochloric acid to the kit.

[0045] Aspect 5: A radiolabeling kit for a prostate-specific membrane antigen-targeting precursor compound according to any one of Aspects 1 to 3, wherein the chelate functional group of the targeting agent is capable of reacting with Ga 3+ / Lu3+ Formation of stable complexes;

[0046] Aspect 6: A radiolabeling kit for prostate-specific membrane antigen-targeting precursor compounds according to any one of Aspects 1 to 3, wherein the acetate is present in a mass between 0.005 g and 1 g;

[0047] Aspect 7: A radiolabeling kit for prostate-specific membrane antigen-targeting precursor compounds according to any one of Aspects 1 to 3, wherein the metal inhibitor is present in micromolar amounts;

[0048] Aspect 8: The radiolabeling kit according to Aspect 1 is used for operation at 25-120°C;

[0049] Aspect 9: The method according to Aspect 1, wherein the acid is hydrochloric acid;

[0050] Aspect 10: The method according to aspect 1 further includes the step of adding hydrochloric acid to the PSMA-targeted precursor compound kit prior to elution;

[0051] Aspect 11: A method for preparing the aforementioned radiolabeled reagent kit for targeting prostate-specific membrane antigens, comprising the following steps:

[0052] a) Preparing or providing a solution comprising at least an appropriate amount of the acetate or a buffer thereof equilibrated to a pH value in the range of 3 to 5 with an acidic pH eluent from a gallium-68 generator / lutetium-177; preparing or providing the prostate-specific membrane antigen-targeting prodrug compound; and preparing or providing the metal inhibitor; and

[0053] b) The solution obtained in step a) by freeze-drying;

[0054] Aspect XII: A method for preparing a radiolabeling kit for the aforementioned prostate-specific membrane antigen-targeting precursor compound, comprising the following steps:

[0055] a) Prepare or provide a solution comprising the chelate-functionalized target agent and the metal inhibitor;

[0056] b) the solution obtained in step a) and c1) adding the acetate as a powder to the lyophilized product obtained in step b), or c2) adding the acetate buffer to the lyophilized product obtained in step b). Attached Figure Description

[0057] Figure 1 This is the mass spectrum of PSMA-HK1.

[0058] Figure 2This is the mass spectrum of PSMA-HK2.

[0059] Figure 3 Mass spectrum of PSMA-HK3

[0060] Figure 4 This is the mass spectrum of PSMA-HK4.

[0061] Figure 5 This is the mass spectrum of PSMA-HK5.

[0062] Figure 6 This is the mass spectrum of PSMA-HK6.

[0063] Figure 7 This is the mass spectrum of PSMA-HK7.

[0064] Figure 8 for[ 68 The radioactive HPLC chromatogram of Ga]Ga-PSMA-HK1~7, where A is [ 68 The radiometric HPLC chromatogram of Ga]Ga-PSMA-HK1, B is [ 68 The radioactive HPLC chromatogram of Ga]Ga-PSMA-HK2, where C is [ 68 The radioactive HPLC chromatogram of Ga]Ga-PSMA-HK3, where D is [ 68 The radiometric HPLC chromatogram of Ga]Ga-PSMA-HK4, E is [ 68 The radioactive HPLC chromatogram of Ga]Ga-PSMA-HK5, F is [ 68 The radioactive HPLC chromatogram of Ga]Ga-PSMA-HK6, where G represents [ 68 Radioactive HPLC chromatogram of Ga]Ga-PSMA-HK7.

[0065] Figure 9 for[ 177 Lu]Lu-PSMA-HK4、[ 177 The radioactive HPLC chromatogram of Lu-PSMA-HK7, where A is [ 177 The radioactive HPLC chromatogram of Lu-PSMA-HK4, B is [ 177 Radioactive HPLC chromatogram of Lu]Lu-PSMA-HK7;

[0066] Figure 10 for[ 68 Ga]Ga-PSMA-HK1~7 and [ 68 Ga]Ga-PSMA-617 in PET images and quantitative analysis of tumor uptake values ​​in LNCaP-bearing mice, where A is [ 68Ga]Ga-PSMA-HK1~7 and [ 68 Ga]Ga-PSMA-617 PET / CT image, B is [ 68 Ga]Ga-PSMA-HK1~7 and [ 68 Quantitative analysis results of tumor uptake values ​​of Ga]Ga-PSMA-617.

[0067] Figure 11 for[ 68 Ga]Ga-PSMA-HK4、[ 68 Ga]Ga-PSMA-HK7 and [ 68 Ga][ 68 The in vitro biodistribution of Ga-PSMA-617 in tumor-bearing mice, where A represents […]. 68 In vitro biodistribution of Ga]Ga-PSMA-HK4, B is [ 68 In vitro biodistribution of Ga]Ga-PSMA-HK7, where C is [ 68 In vitro biological distribution of Ga]Ga-PSMA-617.

[0068] Figure 12 for[ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 The in vitro biodistribution of Lu-PSMA-617 in tumor-bearing mice, where A is [ 177 In vitro biological distribution of Lu]Lu-PSMA-HK4, B is [ 177 In vitro biodistribution of Lu]Lu-PSMA-HK7, C is [ 177 In vitro biological distribution of Lu]Lu-PSMA-617.

[0069] Figure 13 For administration [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7、[ 177 Lu]Lu-PSMA-617、 177 Tumor volume changes in Lu and the control group over 15 days.

[0070] Figure 14 For administration [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7、[ 177 Lu]Lu-PSMA-617、 177 Changes in body weight of mice in Lu and the control group over 15 days. Detailed Implementation

[0071] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention. It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the present invention.

[0072] Furthermore, regarding the numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0073] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar to or equivalent to those described herein may be used in the implementation or testing of this invention.

[0074] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0075] In this invention 177 Lu was purchased through commercial channels, and the radionuclide purity is 98%. 68 Ga from 68 Ge / 68 Ga generator (ITM, Germany). Radiometric high-performance liquid chromatography (HPLC) analysis of all radiotracers was performed using an HPLC system (Shimadzu, Japan).

[0076] All animal experiments used in this invention were approved by the Animal Ethics Committee of Nanjing Medical University. All animals were cared for by humans, and experiments were conducted according to the recommendations of the Laboratory Animal Care and Use Committee. The establishment of the tumor-bearing mouse model followed standard procedures. LNCaP (1×10⁻⁶) was administered subcutaneously. 6 A tumor-bearing Nude-Bulb / c mouse model was established using cells / mice. The tumor volume reached approximately 100-300 mm². 3 Mice were used for PET / CT imaging and biodistribution studies. LNCaP cell lines were maintained in a humidified CO2 incubator (37°C, 5% CO2) and cultured in Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. Subcultures were performed every two days.

[0077] General formula for synthesis:

[0078]

[0079] General method:

[0080] (1) Resin swelling: Add 2-chlorotriphenylmethyl chloride resin (0.44 mmol / g) to DCM (15 mL / g), shake at room temperature for 30 min to swell, and filter out DCM;

[0081] (2) Amino acid linkage: Weigh 5 equivalents of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((((allyloxy)carbonyl)amino)butyric acid or (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((allyloxy)carbonyl)amino)valerate or N-[(9H-fluoren-9-methoxy)carbonyl]-N'-[(2-propenoxy)carbonyl]-L-lysine or (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-7-(((allyloxy)carbonyl)amino)heptanoic acid, dissolve in a small amount of DCM, add 5 equivalents of DIPEA to activate for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0082] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add blocking solution (methanol:DCM:DIPEA=17:2:1, 10mL / g), shake at room temperature for 30min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0083] (4) Fmoc removal, amino conversion and resin linkage: Add 50% morpholine (DMF) solution (15 g / mL) to 0.2 equivalents of compound 2, react with gas for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 1 equivalent of compound 3 and 3 equivalents of DIPEA to 70 mL of dry DCM, add 0.3 equivalents of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 equivalents of resin (Fmoc-removed compound 2) to DCM (4 mL / g), add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0084] (5) Alloc protecting group removal: Take 1 equivalent of compound 5, add 0.15 equivalents of tetra(triphenyl)palladium and 10% morpholine ( / DCM) solution (4 mL / g), react at room temperature for 3 hours to remove the allyloxycarbonyl protecting group, filter out after the reaction and wash the resin 5 times with DMF to obtain compound 6;

[0085] (6) Amino acid linkage: Next, 5 equivalents of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 15 equivalents of HATU, and 40 equivalents of DIPEA were added to 1 equivalent of compound 6. After reacting at room temperature for 2 hours, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0086] (7) Fmoc removal and amino acid linkage: 50% morpholine (DMF) solution (15 g / mL) was added to 1 equivalent of compound 7, and the reaction was carried out under air for 5 min. Morpholine solution was added and the reaction was carried out for another 15 min. The reaction solution was filtered off, and the mixture was washed 6 times with DMF. 5 equivalents of trans-FMOC-4-aminomethylcyclohexanecarboxylic acid or trans-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexane-1-carboxylic acid, 5 equivalents of DIPEA, and 10 equivalents of HATU were added. The mixture was reacted at room temperature for 2 h, filtered off, and the resin was washed. The mixture was then dried to obtain compound 8.

[0087] (8) DOTA-3OtBu linkage: 10 equivalents of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 10 equivalents of DIPEA, and 20 equivalents of HATU were added to 1 equivalent of compound 8. After reacting at room temperature for 12 h, the mixture was filtered out, the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0088] (9) Shearing and purification: Shearing solution (TFA:TIPS:H2O = 95%:2.5%:2.5%, 24 mL / g) was added to compound 9. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, diethyl ether was added, the liquid was discarded after centrifugation, the diethyl ether was removed under reduced pressure, the product was purified by HPLC, and the product was verified by LC-MS.

[0089] Example 1: PSMA-HK1:(3S,11S,15S)-3-(naphth-2-ylmethyl)-1,5,13-trioxo-1-((1r, 4S)-4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)methyl)cyclohexane 2,6,12,14-tetraazaheptadecane-11,15,17-triacetic acid

[0090] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0091] (2) Amino acid linkage: Weigh 397.8 mg of N-[(9H-fluorene-9-methoxy)carbonyl]-N'-[(2-propenoxy)carbonyl]-L-lysine, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0092] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0093] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0094] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0095] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0096] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash with DMF 6 times, add 380 mg of trans-FMOC-4-aminomethylcyclohexanecarboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0097] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0098] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (26.2 mg, yield 12.4%).

[0099] Product validation was performed by LC-MS, MS(ESI) + )m / z 1056.9[M+H] +

[0100] Example 2: PSMA-HK2:(3S,9S,13S)-3-(naphth-2-ylmethyl)-1,5,11-trioxo-1-((1r, 4S)-4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)methyl)cyclohexane 2,6,10,12-tetraazapentadecan-9,13,15-triacetic acid (2,6,10,12-tetraazapentadecan-9,13,15-triacetic acid)

[0101] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0102] (2) Amino acid linkage: Weigh 373.1 mg of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((((allyloxy)carbonyl)amino)butyric acid, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0103] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0104] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0105] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0106] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0107] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash with DMF 6 times, add 380 mg of trans-FMOC-4-aminomethylcyclohexanecarboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0108] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0109] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (24.2 mg, yield 11.8%).

[0110] Product validation was performed by LC-MS, MS(ESI) + )m / z 1028.8[M+H] +

[0111] Example 3: PSMA-HK3:(3S,10S,14S)-3-(naphth-2-ylmethyl)-1,5,12-trioxo-1-((1r, 4S)-4-((2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)methyl)cyclohexane 2,6,11,13-tetraazahexadecane-10,14,16-triacetic acid

[0112] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0113] (2) Amino acid linkage: Weigh 385.4 mg of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((allyloxy)carbonyl)amino)valeric acid, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0114] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0115] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0116] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0117] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0118] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash with DMF 6 times, add 380 mg of trans-FMOC-4-aminomethylcyclohexanecarboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0119] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0120] (9) Shearing and purification: 12 mL of shearing solution was added to the resin and reacted at room temperature for 3 h. The liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (29.6 mg, yield 14.2%).

[0121] Product validation was performed by LC-MS, MS(ESI) + m / z 1042.9 [M+H] +

[0122] Example 4: PSMA-HK4:(3S,11S,15S)-3-(naphth-2-ylmethyl)-1,5,13-trioxo-1-((1r, 4S)-4-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)cyclohexyl)-2, 6,12,14-Tetraazaheptadecane-11,15,17-triacetic acid

[0123] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0124] (2) Amino acid linkage: Weigh 397.8 mg of N-[(9H-fluorene-9-methoxy)carbonyl]-N'-[(2-propenoxy)carbonyl]-L-lysine, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0125] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0126] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0127] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0128] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0129] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash 6 times with DMF, add 346 mg of trans-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexane-1-carboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0130] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0131] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (23.8 mg, yield 11.4%).

[0132] Product validation was performed by LC-MS, MS(ESI) - m / z 1040.9 [MH] --

[0133] Example 5: PSMA-HK5:(3S,9S,13S)-3-(naphth-2-ylmethyl)-1,5,11-trioxo-1-((1r, 4S)-4-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)cyclohexyl)-2, 6,10,12-Tetraazapentadecan-9,13,15-triacetic acid

[0134] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0135] (2) Amino acid linkage: Weigh 373.1 mg of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-((((allyloxy)carbonyl)amino)butyric acid, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0136] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0137] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0138] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0139] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0140] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash 6 times with DMF, add 346 mg of trans-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexane-1-carboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0141] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0142] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (11.0 mg, yield 12.4%).

[0143] Product validation was performed by LC-MS, MS(ESI) + )m / z 1014.7[M+H] +

[0144] Example 6: PSMA-HK6:(3S,10S,14S)-3-(naphth-2-ylmethyl)-1,5,12-trioxo-1-((1r, 4S)-4-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)cyclohexyl)-2, 6,11,13-Tetraazahexadecane-10,14,16-triacetic acid

[0145] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0146] (2) Amino acid linkage: Weigh 385.4 mg of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(((allyloxy)carbonyl)amino)valeric acid, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0147] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0148] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0149] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0150] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0151] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash 6 times with DMF, add 346 mg of trans-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexane-1-carboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0152] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0153] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (27.7 mg, yield 13.5%).

[0154] Product validation was performed by LC-MS, MS(ESI) - m / z 1027.1 [MH] -

[0155] Example 7: PSMA-HK7:(3S,12S,16S)-3-(naphth-2-ylmethyl)-1,5,14-trioxo-1-((1r, 4S)-4-(2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido)cyclohexyl)-2, 6,13,15-Tetraazaoctadecane-12,16,18-triacetic acid

[0156] (1) Resin swelling: Add 1g of 2-chlorotriphenylmethyl chloride resin to 15mL of DCM, shake at room temperature for 30min to swell, and filter out the DCM;

[0157] (2) Amino acid linkage: Weigh 410.1 mg of (S)-2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-7-(((allyloxy)carbonyl)amino)heptanoic acid, dissolve it in 10 mL of DCM, add 612 μL of DIPEA to activate it for 5 min, add the mixed solution to the resin, and react for 2 h under nitrogen protection;

[0158] (3) Resin blocking: Filter out the reaction solution, wash the resin 5 times with DMF, add 10 mL of blocking solution, shake at room temperature for 30 min, filter out the reaction solution, wash with DMF 4 times to obtain compound 2;

[0159] (4) Fmoc removal, amino conversion and resin connection: Weigh 0.2 mmol of compound 2 according to the resin loading, add 8 mL of 50% morpholine (DMF) solution, react under purging for 5 min, continue to add morpholine solution and react for 15 min, filter out the reaction solution, and wash with DMF 6 times; add 295.8 mg of bis(tert-butyl) L-glutamic acid hydrochloride and 0.5 mL of DIPEA to 70 mL of dry DCM, add 101 mg of triphosgene to the solution within 4 h at 0 °C to obtain compound 4, continue to stir and react at 25 °C for 1 h, add 0.2 mmol of resin (compound 2 with Fmoc removed) to 2 mL of DCM, add to compound 4 and react at room temperature for 16 h, filter out after the reaction and wash the resin with DMF 5 times to obtain compound 5;

[0160] (5) Alloc protecting group removal: 0.2 mmol of compound 5 and 35 mg of tetrakis(triphenyl)palladium were added to 2 mL of 10% morpholine (DCM) and reacted at room temperature for 3 hours to remove the allyloxycarbonyl protecting group. After the reaction was completed, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 6.

[0161] (6) Amino acid linkage: Next, 451 mg of Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, 1.6 g of HATU, and 1.4 mL of DIPEA were added to 0.2 mmol of compound 6. After reacting at room temperature for 2 h, the mixture was filtered out and the resin was washed 5 times with DMF to obtain compound 7.

[0162] (7) Fmoc removal and amino acid linkage: Add 8 mL of 50% morpholine (DMF) solution to the resin, purge and react for 5 min, add another 8 mL of morpholine solution and react for 15 min, filter off the reaction solution, wash 6 times with DMF, add 346 mg of trans-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexane-1-carboxylic acid, 180 μL of DIPEA, and 760 mg of HATU, react at room temperature for 2 h, filter out and wash the resin, dry under vacuum to obtain compound 8;

[0163] (8) DOTA-3OtBu linkage: 1.1 g of 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid tritert-butyl ester, 360 μL of DIPEA, and 2.2 g of HATU were added to the resin. After reacting at room temperature for 12 h, the mixture was filtered out and the resin was washed with DMF, dehydrated, and dried to obtain compound 9.

[0164] (9) Shearing and purification: 12 mL of shearing solution was added to the resin. After reacting at room temperature for 3 h, the liquid was filtered, the liquid was removed under reduced pressure, ether was added, the liquid was discarded after centrifugation, the ether was removed under reduced pressure, the product was purified by HPLC, and the product was lyophilized to obtain a white solid (15.6 mg, yield 7.4%).

[0165] Product validation was performed by LC-MS, MS(ESI) + )m / z 1056.7[M+H] +

[0166] The structural formulas of PSMA-HK1 to HK7 are as follows:

[0167]

[0168] Example 1: Evaluation of the synthesis of labeled precursor compounds targeting prostate-specific membrane antigens Synthesized

[0169] The PSMA-HK1 to PSMA-HK7 prepared in Example 1 were analyzed by MS under the following conditions: ion source: ES; nebulizing gas (NEB): 12.00; nitrogen (CUR): 6.00; ion spray voltage (IS): ±4500; temperature (TEM): 0.00; run time: 0.5–1 minute. The mass spectra are shown below. Figure 1-7 The mass spectra are shown for PSMA-HK1 to PSMA-HK7, in that order. The results indicate that the synthesized structure is correct.

[0170] Experimental Example 2: Labeling and Quality Control of PET Tracers Targeting Prostate-Specific Membrane Antigens

[0171] PET tracers targeting prostate-specific membrane antigens [ 68 Synthesis of Ga]Ga-PSMA-HK1~7 (with [ 68 Taking Ga]Ga-PSMA-HK1 as an example, the labeling method for other precursor compounds is the same:

[0172] Take 5 mL of 0.1 mol / L HCl solution and slowly inject it into the germanium-gallium generator for rinsing. When about 1.5 mL of rinsing solution has been discharged, start collecting the leaching liquid (HCl system). 68 GaCl3 milk, approximately 1.3 × 10⁻⁶ 9 Bq, 3.5 mL). Weigh 20 μg PSMA-HK1, add HCl (0.6 mol / L, 1.0 mL, pH = 4) solution, and then add all of the above freshly rinsed solution to the system. 68 The GaCl3 milk solution was reacted at 85℃ for 15 min. After the reaction, the liquid in the system was sterilized by passing it through a 0.22 μm needle filter and collected in a receiving bottle to obtain [ 68 [Ga]Ga-PSMA-HK1. The reaction process is as follows:

[0173]

[0174] Detected by RP-HPLC [ 68 Radiochemical purity of Ga-PSMA-HK1~7 was determined under the following conditions: column: YMC-tria-C18; mobile phase gradient: mobile phase A, 0.1% aqueous trifluoroacetic acid (TFA), mobile phase B, acetonitrile (0.1% TFA); flow time: 15 min, mobile phase B from 20% to 90%; flow rate: 1.0 mL / min. Figure 8 It can be seen from this that, 68 Ga]Ga-PSMA-HK1~[ 68 The radiochemical yield and radiochemical purity of Ga]Ga-PSMA-HK 7 are both greater than 99%;

[0175] Experimental Example 3: PET tracer targeting prostate-specific membrane antigens [ 177 Lu]Lu-PSMA-HK4~7 markers control

[0176] PET tracers targeting prostate-specific membrane antigens [ 177 Synthesis of Lu]Lu-PSMA-HK4~7 (with [ 177 Taking Lu]Lu-PSMA-HK4 as an example, the synthesis methods for other compounds are the same:

[0177] Take 20 μg of PSMA-HK4 and add it to 500 μl of sodium acetate solution (0.25 M). Add... 177 LuCl3 was reacted with an HCl solution (2 mL, 0.05 M, 10 GBq) at 90 °C for 20 min. After the reaction, the liquid in the system was sterilized by passing it through a 0.22 μm needle filter and collected in a receiving bottle to obtain [ 177 Lu]Lu-PSMA-HK1. The reaction process is as follows:

[0178]

[0179] Detected by RP-HPLC [ 177 Lu]Lu-PSMA-HK4 and [ 177 Radiochemical purity of Lu-PSMA-HK7 was determined under the following conditions: column: YMC-tria-C18; mobile phase gradient: mobile phase A, 0.1% aqueous trifluoroacetic acid (TFA), mobile phase B, acetonitrile (0.1% TFA); flow time: 15 min, mobile phase B from 20% to 90%; flow rate: 1.0 mL / min. Figure 9 It can be seen from this that, 177 Lu]Lu-PSMA-HK4 and [ 177 The radiochemical yield and radiochemical purity of Lu]Lu-PSMA-HK 7 are both greater than 99%.

[0180] Example 4 :[ 68 Ga]Ga-PSMA-HK1~7 and [ 68 In vivo PET imaging of Ga-PSMA-617 in tumor-bearing mice Research

[0181] PET scans were performed using Inveon Micro-PET / CT (Siemens Medical Solutions, Knoxville, Munich, Germany). Each mouse was injected via tail vein with 3.7 MBq dissolved in 100 μL saline. 68 Ga]Ga-PSMA-HK1~7 and [ 68 Ga]Ga-PSMA-617. Dynamic PET scanning begins immediately after injection and continues (n=3). In [ 68Static PET images were acquired 1 hour and 2 hours after injection of Ga-PSMA-HK1-7, and tumor uptake values ​​were quantitatively analyzed. Results are shown below. Figure 10 , where A is [ 68 Ga]Ga-PSMA-HK1~7 and [ 68 PET / CT image of Ga-PSMA-617, B is [ 68 Ga]Ga-PSM A-HK1~7 and [ 68 Quantitative analysis results of tumor uptake values ​​of Ga]Ga-PSMA-617. From Figure 10 It can be seen from this that, 68 Ga]Ga-PSMA-HK4 and [ 68 The tumor uptake value of Ga-PSMA-HK7 is higher than that of [ 68 Ga]Ga-PSMA-617 indicates that the radioactive tracer [ 68 Ga]Ga-PSM A-HK4 and [ 68 Ga]Ga-PSMA-HK7 can increase tumor uptake.

[0182] Further measurements were taken. 68 Ga]Ga-PSMA-HK4、[ 68 Ga]Ga-PSMA-HK7 and [ 68 The in vitro biodistribution of Ga-PSMA-617 in tumor-bearing mice is shown in the following results. Figure 11 , where A is [ 68 In vitro biodistribution of Ga]Ga-PSMA-HK4, B is [ 68 In vitro biodistribution of Ga]Ga-PSMA-HK7, where C is [ 68 In vitro biological distribution of Ga]Ga-PSMA-617. Figure 11 It can be seen from this that, 68 Ga]Ga-PSMA-HK4 and [ 68 Ga-PSMA-HK7 was rapidly cleared from the bloodstream within 30 minutes of injection. Similar phenomena were observed in other major organs, including the heart, lungs, pancreas, and bones. Consistent with PET results, the highest renal uptake was observed at all study time points, indicating […]. 68 Ga]Ga-PSMA-HK4 and [ 68 Ga-PSMA-HK7 is primarily excreted by the kidneys. [ 68 Compared to Ga-PSMA-617, the tumor showed a higher radiation signal value, indicating that the radioactive tracer [ 68 Ga]Ga-PSMA-HK4 and [ 68Ga]Ga-PSMA-HK7 compares [ 68 It has a higher tumor uptake value compared to Ga]Ga-PSMA-617.

[0183] Experimental Example 5: [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 Lu]Lu-PSMA-617 in tumor-bearing tissue In vitro biodistribution of mice

[0184] The same method as in Example 4 was used to examine [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 The in vitro biodistribution of Lu-PSMA-617 in tumor-bearing mice is shown in the following results. Figure 12 , where A is [ 177 In vitro biological distribution of Lu]Lu-PSMA-HK4, B is [ 177 In vitro biodistribution of Lu]Lu-PSMA-HK7, C is [ 177 In vitro biodistribution of Lu]Lu-PSMA-617. From... Figure 12 It can be seen from this that, 177 Lu]Lu-PSMA-HK4 and [ 177 Lu-PSMA-HK7 was rapidly cleared from non-target sites such as the blood within 4 hours after injection. Similar phenomena were observed in other major organs, including the heart, lungs, pancreas, and bones. The highest renal uptake was observed at all study time points, indicating […]. 177 Lu]Lu-PSMA-HK4 and [ 177 Lu-PSMA-HK7 is primarily excreted by the kidneys. [ 177 Compared to Lu-PSMA-617, the tumor showed a higher radiation signal value, indicating that the radioactive tracer [ 177 Lu]Lu-PSMA-HK4 and [ 177 Lu]Lu-PSMA-HK7 compared to [ 177 Compared to Lu-PSMA-617, it has higher tumor affinity and uptake value.

[0185] Experimental Example 6: [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 Treatment with Lu-PSMA-617 and safety evaluation

[0186] 18.5 MBq of [method / treatment] was injected into tumor-bearing mice via the tail vein. 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 Following Lu-PSMA-617 administration, changes in tumor volume and body weight were observed in tumor-bearing mice (n=6). Figure 13 It can be seen from the medication [ 177Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7 and [ 177 Following Lu-PSMA-617 administration, tumor volume growth was significantly inhibited, among which [ 177 The tumor volume in mice in the Lu-PSMA-HK4 administration group was smaller than [ 177 Mice in the Lu-PSMA-617-treated group showed that [Lu]Lu-PSMA-617 administration group mice, indicating that [ 177 Lu]Lu-PSMA-HK4 has a stronger tumor-suppressing effect and is more promising in clinical treatment.

[0187] Further determination of drug administration [ 177 Lu]Lu-PSMA-HK4、[ 177 Lu]Lu-PSMA-HK7、[ 177 The changes in body weight of mice in Lu-PSMA-617 and the control group over 15 days are shown in the figure. Figure 14 . Figure 14 The display shows that the drug was administered [ 177 Lu]Lu-PSMA-HK4 and [ 177 Lu]Lu-PSMA-HK7 after [ 177 Compared to Lu-PSMA-617, the mice showed no significant change in body weight. This indicates that the compound involved in this invention is effective for the indicator of body weight change. 177 Similar to Lu-PSMA-617, it has no obvious toxicity.

[0188] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A compound or an acceptable salt thereof that targets a prostate-specific membrane antigen, characterized in that, The general structural formula of the compound is shown in Formula I: Where x is any integer from 0, 1, 2, 3, 4, or 5; m is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; n is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; L1 and L2 are independently selected from straight-chain or branched C1-C4 alkyl groups, -NHCO-, -CH2-O-CH2-, natural amino acids, non-natural amino acids, or one of the following structures, where a is any integer from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10: R is selected from one of the following structures:

2. The compound targeting prostate-specific membrane antigen according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, x is 0 or 1, m is 1, L1 is a C atom, n is 0, 1 or 2, and L2 is a C atom.

3. The compound targeting prostate-specific membrane antigen according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, characterized in that, R is or 4. The compound targeting prostate-specific membrane antigen according to any one of claims 1-2, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

5. A method for preparing the compounds PSMA-HK1, PSMA-HK2, PSMA-HK3, PSMA-HK4, PSMA-HK5, PSMA-HK6, and PSMA-HK7 targeting prostate-specific membrane antigens according to claim 4, comprising the following steps: (1) Using compound 1 as the starting material, compound 1 and DIPEA were added to 2-chloro-triphenylmethyl resin to couple them onto the resin to obtain compound 2. (2) Compound 3 was reacted with DIPEA and triphosgene to obtain compound 4; (3) React compound 2 with compound 4 to obtain compound 5; (4) After the reaction was complete, the resin was filtered out to remove the allyloxycarbonyl protecting group, yielding compound 6. (5) Fmoc-(S)-3-amino-4-(2-naphthyl)-butyric acid, trans-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)cyclohexaneacetic acid or trans-4-(N-fluorenmethoxycarbonylaminomethyl)cyclohexanecarboxylic acid, and tri-tert-butyl 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid were coupled by SPPS method, and purified by shearing resin to obtain A, a PET tracer precursor compound targeting prostate-specific membrane antigen.

6. A radiolabeled compound for diagnosis and treatment, comprising the compound of any one of claims 1-4 that targets prostate-specific membrane antigens, or a radiolabeled compound of an acceptable salt thereof, wherein, The radionuclide is a diagnostic or therapeutic radionuclide selected from... 43 Sc、 44 Sc、 51 Mn, 52 Mn, 64 Cu、 67 Ga、 68 Ga、 90 Y、 89 Zr、 99m Tc, 111 ln、 152 Tb, 155 Tb, 201 Tl、 203 Pb, 212 Pb, 213 Bi、 212 Bi、 225 Ac、 211 At、 186 Re、 67 Cu、 188 Re、 177 Lu、 18 F, 76 Br、 77 Br、 123 I, 124 I, 125 I or 11 At least one of C.

7. A diagnostic radionuclide-labeled compound according to claim 6, wherein, The radionuclide labeling method involves eluting the radionuclide eluent, mixing it with the labeling precursor compound, and adjusting the pH to 4–10 using a buffer solution, wherein the buffer solution is sodium acetate, water, ethanol, phosphate, salt buffer solution or dimethyl sulfoxide; and reacting at 20–110°C for 10 minutes to overnight.

8. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises at least one compound targeting prostate-specific membrane antigen according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or at least one labeled precursor compound prepared according to the method of claim 5, or a pharmaceutically acceptable salt thereof, and at least one diagnostic radionuclide labeled compound according to any one of claims 6 to 7, and optionally a pharmaceutically acceptable diluent, carrier, and / or excipient.

9. Use of the compound targeting prostate-specific membrane antigen according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or at least one diagnostic radionuclide-labeled compound according to any one of claims 6-7, or the pharmaceutical composition according to claim 8, in the preparation of a medicament for the prevention, diagnosis, and / or treatment of prostate-specific membrane antigen-mediated diseases.

10. The use of the diagnostic radionuclide marker compound of claim 6 in the preparation of one or more medicaments or reagents for the diagnosis and / or treatment of one or more tumors, autoimmune diseases, kidney diseases, cardiovascular diseases, inflammation, metabolic / endocrine disorders or neurological diseases.

11. The application according to claim 10, characterized in that, Among them, the radionuclide-labeled compounds used to diagnose and treat diseases include the following pathogenic cells: prostate tumor cells, metastatic prostate tumor cells, lung tumor cells, kidney tumor cells, liver tumor cells, glioblastoma cells, pancreatic tumor cells, bladder tumor cells, sarcoma cells, melanoma cells, breast tumor cells, colon tumor cells, germ cells, pheochromocytoma, esophageal tumor cells, and gastric tumor cells.

12. The application according to claim 10, characterized in that, The diagnostic method is radionuclide imaging, and more preferably includes PET imaging and SPECT imaging. The treatment is selected from radionuclide therapy, and more preferably includes beta radionuclide therapy and alpha radionuclide therapy.

13. A radiolabeling kit for a prostate-specific membrane antigen-targeting compound, comprising: a certain amount of acetate or its salt buffer, to equilibrate the acidic pH eluent from the gallium-68 generator / lutetium-177 accelerator to a pH value in the range of 3 to 5 when eluted in the kit by the gallium-68 generator / lutetium-177 accelerator; The chelating functional group of the prostate-specific membrane antigen-targeting compound is capable of chelating gallium-68 / lutetium-177 under radiolabeling conditions; and a metal inhibitor, which is a co-chelating agent capable of inactivating contaminating metals without interfering with the chelation between gallium-68 / lutetium-177 and the prostate-specific membrane antigen-targeting precursor compound under said radiolabeling conditions, wherein... The metal inhibitor is selected from the group consisting of DOTA, DTPA, monosaccharides, and disaccharides, and the prostate-specific membrane antigen-targeting precursor compound and chelate are derived from claims 1-5.

14. The radiolabeling kit according to claim 13, wherein, The acetate, the prostate-specific membrane antigen-targeting precursor compound, and the metal inhibitor are co-lyophilized.

15. The radiolabeling kit according to claim 13, wherein, The prostate-specific membrane antigen-targeting precursor compound and the metal inhibitor are co-lyophilized, and the acetate or its buffer solution is present alone and may be added subsequently.

16. The radiolabeling kit according to any one of claims 14 to 15, wherein, The acetate or its buffer solution is present at a specific concentration to equilibrate the acidic eluent from the gallium-68 generator / lutetium-177 accelerator to a pH in the range of 3 to 5 when the generator is eluted in the kit after the addition of hydrochloric acid.

17. The radiolabeling kit according to any one of claims 14 to 16, wherein, The chelate functional group of the target agent can react with Ga 3+ / Lu 3+ Forming stable complexes.

18. The radiolabeling kit according to any one of claims 14 to 15, wherein, The acetate is present in quantities between 0.005 g and 1 g.

19. The radiolabeling kit according to any one of claims 14 to 15, wherein, The metal inhibitor is present in micromolar amounts.

20. The radiolabeling kit according to claim 14 is used for operation at 25-120°C.

21. The radiolabeling kit according to claim 14, wherein, The acid in question is hydrochloric acid.

22. The radiolabeling kit of claim 14, further comprising the step of adding hydrochloric acid to the prostate-specific membrane antigen-targeted precursor compound kit prior to elution.

23. A method for preparing a radiolabeling kit for the compound according to claim 1, comprising the following steps: a) Preparing or providing a solution comprising at least an appropriate amount of the acetate or a buffer thereof equilibrated to a pH value in the range of 3 to 5 with an acidic pH eluent from a gallium-68 generator / lutetium-177; preparing or providing the prostate-specific membrane antigen-targeting prodrug compound; and preparing or providing the metal inhibitor; and b) The solution obtained in step a) by freeze-drying.

24. A method for preparing the compound according to claim 1, comprising the following steps: a) Prepare or provide a solution comprising the chelate-functionalized target agent and the metal inhibitor; b) the solution obtained in step a) and c1) adding the acetate as a powder to the lyophilized product obtained in step b), or c2) adding the acetate buffer to the lyophilized product obtained in step b).

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