Prostate-specific membrane antigen inhibitors, radiolabeled versions thereof, and methods of making and using

Abstract

The application discloses a prostate specific membrane antigen inhibitor, a radioactive marker thereof, and a preparation method and application thereof, and belongs to the technical field of biological medicines. The prostate specific membrane antigen inhibitor is shown as formula I. The radioactive marker of the prostate specific membrane antigen inhibitor is shown as formula II. The application discloses a preparation method of the compound of formula I. The preparation method of the compound of formula II comprises the following steps: reacting the compound of formula I with a radioactive metal salt to generate the radioactive marker of the compound of formula I, i.e. the compound of formula II. The application discloses application of the compound of formula I or the radioactive marker of the prostate specific membrane antigen inhibitor of formula II in preparation of a prostate cancer diagnosis reagent / drug or / and a therapeutic drug. The radioactive marker of the compound of formula I has high tumor uptake and an enhanced tumor-to-background ratio in a prostate cancer animal model.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and particularly relates to a prostate specific membrane antigen inhibitor, a radioactive marker thereof, and a preparation method and application thereof. BACKGROUND

[0002] Prostate cancer is an epithelial malignant tumor occurring in the prostate and is the most common malignant tumor of the male urogenital system. The incidence of prostate cancer is extremely high in elderly men, and the incidence rate is on the level before the age of 50, and the incidence rate gradually increases with age, and 80% of cases occur in men over 65 years old. The global incidence of prostate cancer continues to rise, and according to statistics, there were nearly 1.3 million new cases and 359,000 deaths worldwide in 2018, accounting for 13.5% of the incidence of male malignant tumors, ranking second; accounting for 6.7% of the mortality rate of male malignant tumors, ranking fifth.

[0003] In the prior art, a drug based on prostate cancer specific membrane antigen (PSMA) as a probe for positron emission tomography (PET) imaging is used for the diagnosis and treatment of prostate cancer. The applicant's prior application CN115010629A discloses a prostate specific membrane antigen inhibitor, a radionuclide marker thereof, and a preparation method and application thereof. The prostate specific membrane antigen inhibitor thereof is a polyamine carboxylic acid short peptide, and the molecular skeleton thereof is composed of a polyamine polyacid, a fatty amine linking group, and a hexanedioic acid monamide. The lysine-monamide-hexanedioic acid is a PSMA targeting part, the polyamine carboxylic acid is a chelating group for a radionuclide, and the fatty amine is a linking group. The positron marker labeling product is stable in a PBS buffer solution and fetal bovine serum, the positron marker has high tumor uptake and enhanced tumor-to-background ratio in a prostate cancer animal model, and can be used in the fields of diagnosis, staging, efficacy evaluation, and treatment of prostate cancer. However, more PSMA small molecule inhibitors with novel structures, high uptake rates, and reasonable metabolic properties are still needed in clinical practice. SUMMARY

[0004] One of the purposes of the present application is to provide a prostate specific membrane antigen inhibitor shown in formula I, which has a novel structure, stable physical and chemical properties, and can be used in the fields of diagnosis, staging, efficacy evaluation, and treatment of prostate cancer.

[0005] The second purpose of the present application is to provide a radioactive marker of the prostate specific membrane antigen inhibitor, and the structure of the radioactive marker is shown in formula II. The radioactive marker has a high labeling rate, high cell uptake and internalization, long drug retention time in tumors, clear imaging of prostate cancer, and can be used in the fields of diagnosis, staging, efficacy evaluation, and treatment of prostate cancer.

[0006] The third purpose of the present application is to provide a preparation method of the prostate specific membrane antigen inhibitor shown in formula I.

[0007] The fourth object of the present application is to provide a method for preparing the radionuclide marker shown in formula II.

[0008] The fifth object of the present application is to provide an application of the prostate-specific membrane antigen inhibitor shown in formula I.

[0009] The sixth object of the present application is to provide an application of the radionuclide marker pair shown in formula II.

[0010] To achieve the above object, the technical scheme adopted by the present application is as follows:

[0011] The present application discloses a prostate-specific membrane antigen inhibitor shown in formula I,

[0012]

[0013]

[0014]

[0015] X is alkyl or halogen;

[0016]

[0017] When a = b = e = 0, d = 1, and c = an integer from 1 to 8;

[0018] When c = e = 0, a = d = 1, and b = an integer from 1 to 8;

[0019] When c = d = 0, a = e = 1, and b = an integer from 1 to 8.

[0020] In some embodiments of the present application, X is C1-C4 alkyl or halogen;

[0021] Preferably, the halogen is I;

[0022] Preferably, the alkyl is methyl.

[0023] The compound of formula I of the present application is a polyamine carboxylic acid short peptide, and the molecular skeleton is composed of a polyamine polyacid, a fatty amine linking group, and a single amide adipic acid. The lysine-single amide-adipic acid is a PSMA targeting part, the polyamine carboxylic acid is 68 Ga, 177 Lu, 64 Cu, 18 F and 225 Ac chelating group, and the fatty amine or amide or polyethylene glycol is a linking group. The structure is novel, the physicochemical properties are stable, and it is used in the fields of diagnosis, staging, efficacy evaluation, and treatment of prostate cancer.

[0024] The above-mentioned prostate-specific membrane antigen inhibitor of the present application is a radiolabeling agent, and its structure is shown in formula II:

[0025]

[0026]

[0027] wherein M is a radionuclide;

[0028] R1, A, B, C, D, E, X, a, b, c, d, and e are defined as above.

[0029] In some embodiments of the present application, when R3= DOTA-M, M= 68 Ga, 177 Lu, 225 Ac or 64 Cu;

[0030] when R3= DOTA-Ga-M, M= 68 Ga, 177 Lu, 225 Ac or 64 Cu;

[0031] when R3= NOTA-M, M= 68 Ga or 18 F;

[0032] when R3= DATA-M, M= 68 Ga, 177 Lu, 225 Ac or 64 Cu;

[0033] when R3= DATA-M-C, C= 68 Ga, 177 Lu, 225 Ac or 64 Cu;

[0034] when R3= HBED-CC-M, M= 68 Ga.

[0035] The compound of formula II of the present application is a polyamine carboxylic acid short peptide radionuclide labeling agent, and the molecular skeleton is composed of a polyamine carboxylic acid labeled with a radioactive metal M or C, a fatty amine / amide / polyethylene glycol linker, lysine, and a monoamide adipic acid. Among them, the lysine-monoamide adipic acid is the PSMA targeting part, the radionuclide labeling is the imaging functional module or the therapeutic functional module of the drug, and the fatty amine / amide / polyethylene glycol is the linker. When the radionuclide = 68 Ga, 18 F or64 Cu, radionuclide labels the imaging functional module of the drug; when radionuclide = 177 Lu or 225 Ac, radionuclide labels the therapeutic functional module of the drug. The label structure is novel, the physical and chemical properties are stable, the labeling rate is high, the affinity to cells with high expression of PSMA is high, the imaging of tumor-bearing mice is clear, the tumor uptake is good, and the retention time in the tumor is long, and can be used for the diagnosis, staging, efficacy evaluation and other imaging and treatment fields of prostate cancer.

[0036] The preparation method of the compound of formula I disclosed in the application comprises the following steps:

[0037] Step 1. Compound b6 and b7 undergo nucleophilic substitution reaction to generate compound b8;

[0038] Step 2. Compound b8 is deprotected to generate compound b9;

[0039] Step 3. Compound b9 and b10 undergo polypeptide coupling reaction to generate compound b11;

[0040] Step 4. Compound b11 is deprotected to generate compound of formula I;

[0041] The reaction route is as follows:

[0042]

[0043] ; wherein, R1, R2, A, B, C, D, E, X, a, b, c, d, e are as defined above.

[0044] In some embodiments of the application, in step 1, compound b6 and compound b7 are coupled to generate compound b8 in the presence of a basic solvent and a condensing agent; preferably, the molar ratio of compound b7 to compound b6 is 1.0-2.0;

[0045] Or / and in step 2, compound b8 is reduced with a reducing agent in a solvent to generate compound b9;

[0046] Or / and in step 3, compound b9 and compound b10 are coupled to generate compound b11 in the presence of a basic solvent and a condensing agent; preferably, the molar ratio of compound b9 to compound b10 is 1.0-2.0;

[0047] Or / and in step 4, compound b11 is deprotected in an acidic solvent to generate compound of formula I;

[0048] Preferably, the base in step 1 and step 3 comprises at least one of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, cyclohexylamine; further preferably, the molar ratio of the base to b6 or b9 is 1.0-5.0;

[0049] Preferably, the condensing agent used in the coupling reaction in step 1 and step 3 comprises at least one of HATU, HBTU, HOBT, DCC, EDCI;

[0050] The solvent in step 1-step 4 is an aprotic polar solvent, further preferably comprising at least one of dichloromethane, chloroform, tetrahydrofuran, 1,4-dioxane, acetonitrile;

[0051] The reaction temperature in step 1-step 4 is 0-60℃, and the reaction time is 1-24 hours.

[0052] In some embodiments of the present application, the preparation method of the compound 6 comprises the following steps:

[0053] S1-1. Mannich reaction of compound b1 and b2-1 to generate compound b3-1;

[0054] S1-2. Nucleophilic substitution reaction of compound b3-1 and b4 to generate compound b5-1;

[0055] S1-3. Azide reduction reaction of compound b5-1 to generate compound b6;

[0056] The reaction route is as follows:

[0057]

[0058] wherein R1, A, B, C, a, b, c, and the definitions of are the same as above, and n is an integer of 1-6;

[0059] Preferably, in S1-1, Mannich reaction of compound b1, compound b2-1 and CH3BNNa in an organic solvent generates compound b3-1; preferably, the molar ratio of compound b1 to compound b2-1 is 1.0-3.0; preferably, the molar ratio of compound b1 to CH3BNNa is 1.0-3.0; preferably, the organic solvent used in S1-1 is a polar solvent; further preferably, it comprises any one or several of dimethylformamide, methanol, ethanol, water, formic acid, acetic acid, hydrochloric acid; preferably, the reaction temperature in S1-1 is 0-80℃, and the reaction time is 4-24 hours;

[0060] Preferably, in the S1-2, the compound b3-1 is subjected to nucleophilic substitution reaction with b4 in a basic organic solvent to generate compound b5-1; preferably, the molar ratio of compound b3-1 to b4 is 1.0-3.5; preferably, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, cyclohexylamine, and the molar ratio of the base to compound b3-1 is 1.0-5.0; preferably, the organic solvent in the S1-2 is an aprotic polar solvent, and further preferably, includes any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile; preferably, the reaction temperature of the S1-2 is 0-60℃, and the reaction time is 4-48 hours.

[0061] Preferably, in the S1-3, the compound b5-1 is subjected to reduction reaction with a reducing agent in a basic organic solvent to generate b6; preferably, the reducing agent is Pd / C and hydrogen or triphenylphosphine, and the amount of Pd / C is 1.25-20.50% of the molar amount of compound b5-1, the amount of hydrogen is 1.0-20.0% of the molar amount of compound b5-1, and the amount of triphenylphosphine is 1.0-5.0 times of the molar amount of compound b5-1; preferably, the organic solvent is a polar solvent, and further preferably, includes any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile, water, methanol, and ethanol; preferably, the reaction temperature of the S1-3 is 0-90℃, and the reaction time is 1-20 hours.

[0062] In some embodiments of the present application, the method for preparing the compound 6 comprises the following steps:

[0063] S2-1. Mannich reaction of compound b1 with b2-2 to generate compound b3-2;

[0064] S2-2. Nucleophilic substitution reaction of compound b3-2 with b4 to generate compound b5-2-1;

[0065] S2-3. Compound b5-2-1 is subjected to deprotection to generate compound b5-2-2;

[0066] S2-4. Peptide coupling reaction of compound b5-2-2 with b5-2-3 to generate compound b5-2-4;

[0067] S2-5. Compound b5-2-4 is subjected to deprotection to generate compound b6;

[0068] The reaction route is shown as follows:

[0069]

[0070] wherein R1, A, B, C, a, b, c, are as defined above, and n is an integer from 1 to 6;

[0071] Preferably, in S2-1, compound b1 is subjected to Mannich reaction with compound b2-2 and NaBH4 in an organic solvent to generate compound b3-2; preferably, the molar ratio of compound b1 to compound b2-2 is 1.0-3.0; preferably, the molar ratio of compound b1 to NaBH4 is 1.0-3.0; preferably, the organic solvent is a polar organic solvent; further preferably, it includes any one or several of dimethylformamide, methanol, ethanol, water, formic acid, acetic acid, hydrochloric acid; preferably, the reaction temperature of S2-1 is 0-80℃, and the reaction time is 4-24 hours;

[0072] Preferably, in S2-2, compound b3-2 is subjected to nucleophilic substitution reaction with b4 in a basic organic solvent to generate compound b5-2-1; preferably, the molar ratio of compound b3-2 to compound b4 is 1.0-3.5; preferably, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, cyclohexylamine, and the molar ratio of the base to compound b3-2 is 1.0-5.0; preferably, the organic solvent in S2-2 is an aprotic polar solvent; further preferably, it includes any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile; preferably, the reaction temperature in S2-2 is 0-60℃, and the reaction time is 4-48 hours;

[0073] Preferably, in S2-3, compound b5-2-1 is deprotected under basic solvent conditions to generate compound b5-2-2; preferably, the base is one or more of lithium hydroxide, sodium hydroxide, potassium hydroxide, barium hydroxide, and the molar ratio of the base to compound b5-2-1 is 1.0-5.0; preferably, the solvent is a protic polar solvent, and further preferably, it includes any one or several of water, methanol, ethanol, isopropanol, n-butanol; preferably, the reaction temperature in S2-3 is 0-90℃, and the reaction time is 1-20 hours;

[0074] Preferably, in the S2-4, the compound b5-2-2 is coupled with the compound b5-2-3 in the presence of a basic solvent and a condensing agent to generate the compound b5-2-4; preferably, the molar ratio of the compound b5-2-3 to the compound b5-2-2 is 1.0-3.0; preferably, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, cyclohexylamine, and the molar ratio of the base to the compound b5-2-2 is 1.0-5.0; preferably, the coupling agent used is one or more of HATU, HBTU, HOBT, DCC, EDCI, and the molar ratio of the coupling agent to the compound b5-2-2 is 1.0-5.0; preferably, the solvent is an aprotic polar solvent, and further preferably, the solvent comprises any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile; preferably, the reaction temperature of S2-4 is 0-60℃, and the reaction time is 1-24 hours.

[0075] Preferably, in the S2-5, the compound b5-2-4 is deprotected in a basic solvent to generate the compound b6; preferably, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, cyclohexylamine, and the molar ratio of the base to the compound b5-2-4 is 1.0-5.0; preferably, the solvent is an aprotic polar solvent, and further preferably, the solvent comprises any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile; preferably, the reaction temperature of S2-5 is 0-90℃, and the reaction time is 1-24 hours.

[0076] The application discloses a preparation method of a compound of formula II, comprising the following steps: reacting a compound of formula I with a radioactive metal salt to generate a radioactive marker of the compound of formula I, i.e., a compound of formula II, and the reaction formula is as follows:

[0077]

[0078] wherein R1, R 2、 R 3、 A, B, C, D, E, a, b, c, d, and e are defined as above.

[0079] In some embodiments of the application, in the preparation method of the radioactive nuclide marker of the compound of formula I, the pH value of the reaction system is 3.5-10.0, the reaction temperature is 25-95℃, and the reaction time is 5-60 min.

[0080] Preferably, the reaction system further comprises a stabilizer, and further preferably, the stabilizer is selected from any one or several of ethanol, vitamin C, tyrosine, cysteine, serine, and gentisic acid.

[0081] This invention adjusts the pH value by adding a buffer solution to the reaction system. The buffer solution is selected from sodium acetate / acetic acid system, ammonium acetate / acetic acid system, sodium acetate / hydrochloric acid system, HEPES system or Tris system.

[0082] In some embodiments of the present invention, in the method for preparing the radionuclide label of the compound of formula I, the reaction solvent is one or any combination of two or three solvents, namely buffer solution, pure water, and 0.85% to 0.9% physiological saline.

[0083] The use of the compounds of Formula I or radiolabeled prostate-specific membrane antigen inhibitors of Formula II disclosed in this invention in the preparation of diagnostic reagents / drugs and / or therapeutic drugs for prostate cancer.

[0084] The Chinese names corresponding to the English abbreviations of the compounds or groups described in this invention are:

[0085] HBTU: Benzotriazole-N,N,N',N'-Tetramethylurea hexafluorophosphate; HATU: 2-(7-azabenzotriazole)-N,N,N',N'-Tetramethylurea hexafluorophosphate; HOBT: 1-Hydroxybenzotriazole; DCC: N,N'-Dicyclohexylcarbodiimide; EDCI: 1-Ethyl-(3-Dimethylaminopropyl)carbodiimide hydrochloride; tBu: Tert-butyl; TEA: Triethylamine; DEA: Diethanolamine; TFA: Trifluoroacetic acid; DMF: N,N-Dimethylformamide; EA: Ethyl acetate; EtOH: Ethanol; THF: Tetrahydrofuran.

[0086] Compared with the prior art, the present invention has the following beneficial effects:

[0087] The polyamine carboxylic acid short peptide compound of formula I synthesized in this invention is easy to prepare, has a short preparation cycle, and the radiolabeling process is simple and mild. The labeled product is stable in PBS buffer solution and fetal bovine serum. The positron-emitting label has high tumor uptake and enhanced tumor-to-background ratio in prostate cancer animal models, and can be used in the clinical diagnosis, staging, efficacy evaluation and treatment of prostate cancer. Attached Figure Description

[0088] Appendix Figure 1 The image shows the LC-MS mass spectrum of Equation I-1-5-BD-WJ-DOTA.

[0089] Appendix Figure 2 The image shows the LC-MS mass spectrum of Equation I-2-4-BD-WJ-DOTA.

[0090] Appendix Figure 3 The image shows the LC-MS mass spectrum of Equation I-2-4-PN-WJ-DOTA.

[0091] AppendixFigure 4 for 68 Radioactive high-performance liquid chromatogram of Ga-type ⅠI-1-5-BD-WJ-DOTA.

[0092] Appendix Figure 5 for 68 Radioactive high-performance liquid chromatography chromatogram of Ga-type ⅠI-2-4-BD-WJ-DOTA.

[0093] Appendix Figure 6 for 68 Radioactive high-performance liquid chromatography chromatogram of Ga-type ⅠI-2-4-PN-WJ-DOTA.

[0094] Appendix Figure 7 for 177 Radioactive high-performance liquid chromatogram of Lu-type ⅠI-1-5-BD-WJ-DOTA.

[0095] Appendix Figure 8 for 177 Radioactive high-performance liquid chromatography chromatogram of Lu-type ⅠI-2-4-BD-WJ-DOTA.

[0096] Appendix Figure 9 for 177 Radioactive high-performance liquid chromatogram of Lu-type ⅠI-2-4-PN-WJ-DOTA.

[0097] Appendix Figure 10 for 68 Radiometric high-performance liquid chromatography (HPLC) chromatogram of Ga-type ⅠI-1-5-BD-WJ-DOTA in fetal bovine serum (120 minutes).

[0098] Appendix Figure 11 for 68 Radiometric high-performance liquid chromatography (HPLC) chromatogram of Ga-type II-2-4-BD-WJ-DOTA in fetal bovine serum (120 min)

[0099] Appendix Figure 12 for 68 Radiometric high-performance liquid chromatography (HPLC) chromatogram of Ga-type II-2-4-PN-WJ-DOTA in fetal bovine serum (120 min).

[0100] Appendix Figure 13 for 68 PET / CT images of Ga-type Ⅱ-1-5-BD-WJ-DOTA, where the animal model is a NOD / SCID double-deficient mouse inoculated with LNCaP, and the imaging times are 10 minutes, 30 minutes, 60 minutes and 120 minutes after drug administration (from left to right).

[0101] Appendix Figure 14 for 68Ga-PET / CT imaging of Formula I I-2-4-BD-WJ-DOTA, wherein the animal model is LNCaP inoculated NOD / SCID double-deficient mice, and the imaging time is 10 min, 30 min, 60 min and 120 min (from left to right) after administration.

[0102] Figure 8 Figure 15 Figure 9 68 Ga-PET / CT imaging of Formula I I-2-4-PN-WJ-DOTA, wherein the animal model is LNCaP inoculated NOD / SCID double-deficient mice, and the imaging time is 10 min, 30 min, 60 min and 120 min (from left to right) after administration.

[0103] Figure 10 Figure 16 Figure 11 177 Lu-SPECT / CT imaging of Formula I I-1-5-BD-WJ-DOTA, wherein the animal model is LNCaP inoculated NOD / SCID double-deficient mice, and the imaging time is 1 h, 4 h, 24 h, 48 h and 96 h (from left to right) after administration.

[0104] Figure 12 Figure 17 Figure 13 177 Lu-SPECT / CT imaging of Formula I I-2-4-BD-WJ-DOTA, wherein the animal model is LNCaP inoculated NOD / SCID double-deficient mice, and the imaging time is 1 h, 4 h, 24 h, 48 h and 96 h (from left to right) after administration.

[0105] Figure 14 Figure 18 Figure 15 177 Lu-SPECT / CT imaging of Formula I I-2-4-PN-WJ-DOTA, wherein the animal model is LNCaP inoculated NOD / SCID double-deficient mice, and the imaging time is 1 h, 4 h, 24 h, 48 h and 96 h (from left to right) after administration.

[0106] Figure 16 Figure 19 Figure 17 177 Figure of the results of the in vivo distribution of Lu-PSMA-617.

[0107] Figure 18 Figure 20 Figure 19 177 Figure of the results of the in vivo distribution of Lu-Flu-1.

[0108] Figure 20 Figure 21 Figure 21 177 Figure of the results of the in vivo distribution of Lu-Formula II-1-5-BD-WJ-DOTA;

[0109] Figure 22 Figure 22For example 15 177 Figure showing the in vivo distribution of Lu-type II-2-4-BD-WJ-DOTA;

[0110] Appendix Figure 23 For example 15 177 Figure showing the results of the in vivo distribution of Lu-type II-2-4-PN-WJ-DOTA;

[0111] Appendix Figure 24 At a dose of 37 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H).

[0112] Appendix Figure 25 At a dose of 18.5 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H).

[0113] Appendix Figure 26 At a dose of 7.4 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H). Detailed Implementation

[0114] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0115] Example 1-1

[0116] This embodiment discloses the synthesis of compound b3-1-3 (i.e., in compound b3-1, n=3), and its reaction formula is as follows:

[0117]

[0118] Specifically, compound b1 (1.224 g, 2.513 mmol) and b2-1-3 (335 mg, 2.639 mmol) were dissolved in methanol (40 mL), stirred at 0 °C for 30 min. CH3BNNa (241 mg, 3.77 mmol) was added to the above solution and stirred at room temperature for 4 h. The reaction solution was poured into ice water and extracted with ethyl acetate, washed with saturated brine for 3 times, dried over anhydrous sodium sulfate overnight. The organic phase was filtered and the organic solvent was removed by rotary evaporation. The product was purified by column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain yellow oily liquid b3-1-3, a total of 1.15 g, yield: 55.3%.

[0119] Example 2-1

[0120] This example discloses the synthesis of compound b5-1-3-WJ (i.e. compound b5-1, n = 3, R1 = WJ), whose reaction formula is as follows:

[0121]

[0122] Specifically, b3-1-3 (220 mg, 0.367 mmol) and b4-WJ (101 mg, 0.441 mmol) were dissolved in dichloromethane (3 mL), and then triethylamine (75 mg, 0.735 mmol) was added, and stirred at room temperature for 12 h. After the reaction was completed, it was washed with saturated potassium carbonate solution for 3 times, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate overnight. The organic phase was rotary evaporated to remove the organic solvent. The product was purified by column chromatography (petroleum ether / ethyl acetate = 4 / 1) to obtain yellow oily liquid product b5-1-3-WJ, a total of 187 mg, yield 64.4%.

[0123] Example 3-1

[0124] This example discloses the synthesis of compound b6-1-5-WJ (i.e. compound b6, a = b = 0, c = 5, R1 = WJ), whose reaction formula is as follows:

[0125]

[0126] Specifically, the reactants b5-1-3-WJ (158 mg, 0.200 mmol) and triphenylphosphine (90 mg, 0.343 mmol) were dissolved in tetrahydrofuran (2 ml) and water (1 ml), and stirred at room temperature for 5 h. After the reaction was completed, the reaction solution was poured into ice water and extracted with dichloromethane, washed with saturated brine for 3 times, and the organic phase was dried over anhydrous sodium sulfate overnight. The organic phase was filtered and the organic solvent was removed by rotary evaporation. Column chromatography (dichloromethane / methanol = 15 / 1) was used to obtain yellow oily liquid product b6-1-5-WJ, a total of 135 mg, yield: 88.4%.

[0127] Example 1-2

[0128] This example discloses the synthesis of compound b3-2, the reaction formula of which is as follows:

[0129]

[0130] Specifically, compound b1 (1.15 g, 2.358 mmol) and b2-2 (250 mg, 2.83 mmol) were dissolved in methanol (10 mL), stirred at 0 °C for 30 min. NaBH4 (134 mg, 3.537 mmol) was added to the above solution and stirred at room temperature for 6 h. The reaction was quenched with saturated sodium bicarbonate solution and extracted with ethyl acetate, washed with saturated brine for 3 times, dried over anhydrous sodium sulfate. The organic phase was separated by filtration and the organic solvent was removed by rotary evaporation. The product was purified by column chromatography (dichloromethane / methanol = 30 / 1) to obtain transparent colorless oily liquid b3-2, total 877 mg, yield: 66.7%.

[0131] Example 2-2

[0132] This example discloses the synthesis of compound b5-2-1-WJ (i.e. in compound b5-2-1, R1 = WJ), the reaction formula of which is as follows:

[0133]

[0134] Specifically, b3-2 (430 mg, 0.768 mmol) and b4-WJ (210 mg, 0.922 mmol) were dissolved in dichloromethane (4 mL), and triethylamine (94 mg, 0.922 mmol) was added, stirred at room temperature for 10 h. After the reaction was completed, it was washed with saturated potassium carbonate solution for 3 times, extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate overnight. The organic phase was rotary evaporated to remove the organic solvent. The product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain yellow oily liquid product b5-2-1-WJ, total 420 mg, yield 72.7%.

[0135] Example 3-2-1

[0136] This example discloses the synthesis of compound b5-2-2-WJ (i.e. in compound b5-2-2, R1 = WJ), the reaction formula of which is as follows:

[0137]

[0138] Specifically, b5-2-1-WJ (300 mg, 0.399 mmol) was dissolved in methanol (5 ml), and 5 ml of aqueous lithium hydroxide solution (20 mg / ml) was added, and stirred at room temperature for 10 hours. After the reaction was completed, the methanol was removed by rotary evaporation under reduced pressure, the reaction solution was diluted with water, the pH was adjusted to 5-6 with 1M hydrochloric acid, and the reaction solution was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate. The organic phase was rotary evaporated to remove the organic solvent, and the product was purified by column chromatography (dichloromethane / methanol = 10 / 1) to obtain a white solid crude product b5-2-2-WJ, a total of 187 mg, a yield of 63.6%.

[0139] Example 3-2-2

[0140] This example discloses the synthesis of compound b5-2-4-4-WJ (i.e., compound b5-2-4, n = 4, R1 = WJ), the reaction formula of which is as follows:

[0141]

[0142] Specifically, b5-2-2-WJ (150 mg, 0.203 mmol) was dissolved in DMF (2 mL), HATU (93 mg, 0.244 mmol) and diisopropylethylamine (32 mg, 0.244 mmol) were added, and stirred at room temperature for 30 minutes, then b5-2-3-4 (112 mg, 0.244 mmol) was added, and stirred at room temperature for 8 hours. The reaction solution was poured into ice water, extracted with dichloromethane, washed with saturated brine 3 times, and the organic phase was collected and dried over anhydrous sodium sulfate overnight. After filtration, the organic solvent was removed by rotary evaporation, and the product was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain a colorless oily liquid product b5-2-4-4-WJ, a total of 124 mg, a yield of 51.7%.

[0143] Example 3-2-3

[0144] This example discloses the synthesis of compound b6-2-4-WJ (i.e., compound b6, c = 0, a = 1, b = 4, R1 = WJ), the reaction formula of which is as follows:

[0145]

[0146] b5-2-4-4-WJ (110 mg, 0.093 mmol) was dissolved in dichloromethane (2 ml), and 25% diethanolamine was added, and the reaction was stirred at room temperature overnight. After the reaction was completed, the organic solvent was removed by rotary evaporation, and the reaction product was washed with n-hexane repeatedly several times, and the product was purified by column chromatography (dichloromethane / methanol = 8 / 1) to obtain a colorless viscous liquid product b6-2-4-WJ, a total of 42 mg, a yield of 47.2%.

[0147] Example 4-1

[0148] This example discloses the synthesis of compound b8-1-5-BD-WJ (i.e. in compound b8, a = b = e = 0, d = 1, X = CH3, c = 5, R1= WJ), whose reaction formula is as follows:

[0149]

[0150] Specifically, b7-BD (200 mg, 0.425 mmol) was dissolved in DMF (3 mL), EDCI (66 mg, 0.489 mmol), HOBT (94 mg, 0.489 mmol) and TEA (65.9 mg, 0.652 mmol) were added, and after stirring at room temperature for 30 minutes, b6-1-5-WJ (250 mg, 0.326 mmol) was added, and stirring was performed at room temperature for 10 hours. After the reaction was completed, ice water was added for quenching, and the organic phase was extracted with ethyl acetate, washed with saturated brine for 3 times, and finally the organic phase was collected and dried over anhydrous sodium sulfate overnight. After filtration, the organic solvent was removed by rotary evaporation, and the product was purified by column chromatography (dichloromethane / methanol = 40 / 1) to obtain colorless oily liquid product b8-1-5-BD-WJ, a total of 190 mg, with a yield of 47.8%.

[0151] Example 4-2

[0152] This example discloses the synthesis of compound b8-1-5-BID-WJ (i.e. in compound b8, a = b = e = 0, d = 1, X = I, c = 5, R1= WJ), whose reaction formula is as follows:

[0153]

[0154] Specifically, b7-BID (248 mg, 0.425 mmol) was dissolved in DMF (3 mL), EDCI (66 mg, 0.489 mmol), HOBT (94 mg, 0.489 mmol) and TEA (66 mg, 0.652 mmol) were added, and after stirring at room temperature for 30 minutes, b6-1-5-WJ (250 mg, 0.326 mmol) was added, and stirring was performed at room temperature for 10 hours. After the reaction was completed, ice water was added for quenching, and the organic phase was extracted with ethyl acetate, washed with saturated brine for 3 times, and finally the organic phase was collected and dried over anhydrous sodium sulfate overnight. After filtration, the organic solvent was removed by rotary evaporation, and the product was purified by column chromatography (dichloromethane / methanol = 40 / 1) to obtain colorless oily liquid product b8-1-5-BID-WJ, a total of 233 mg, with a yield of 53.6%.

[0155] Example 4-3

[0156] This example discloses the synthesis of compound b8-2-4-PN-WJ (i.e. in compound b8, a = e = 1, c = d = 0, b = 4, R1= WJ), whose reaction formula is as follows:

[0157]

[0158] b7-PN (233 mg, 0.425 mmol) was dissolved in DMF (3 mL), EDCI (66 mg, 0.489 mmol), HOBT (94 mg, 0.489 mmol) and TEA (66 mg, 0.652 mmol) were added, after stirring at room temperature for 30 minutes, b6-2-4-WJ (269 mg, 0.326 mmol) was added, and stirring was carried out at room temperature for 10 hours. After the reaction was completed, ice water was added for quenching, and the organic phase was extracted with ethyl acetate, washed with saturated brine for 3 times, and finally the organic phase was collected, dried with anhydrous sodium sulfate overnight. After filtration, the organic solvent was removed by rotary evaporation, and the product was purified by column chromatography (dichloromethane / methanol = 35 / 1) to obtain colorless oily liquid product b8-2-4-PN-WJ, a total of 318.2 mg, with a yield of 74.1%.

[0159] Example 4-4

[0160] This example discloses the synthesis of compound b8-2-4-BD-WJ (i.e. in compound b8, a = d = 1, X = CH3, b = 4, c = e = 0, R1= WJ), whose reaction formula is as follows:

[0161]

[0162] Specifically, b7-BD (200 mg, 0.425 mmol) was dissolved in DMF (3 mL), EDCI (66 mg, 0.489 mmol), HOBT (94 mg, 0.489 mmol) and TEA (66 mg, 0.652 mmol) were added, after stirring at room temperature for 30 minutes, b6-2-4-WJ (269 mg, 0.326 mmol) was added, and stirring was carried out at room temperature for 10 hours. After the reaction was completed, ice water was added for quenching, and the organic phase was extracted with ethyl acetate, washed with saturated brine for 3 times, and finally the organic phase was collected, dried with anhydrous sodium sulfate overnight. After filtration, the organic solvent was removed by rotary evaporation, and the product was purified by column chromatography (dichloromethane / methanol = 35 / 1) to obtain colorless oily liquid product b8-2-4-BD-WJ, a total of 301 mg, with a yield of 65.5%.

[0163] Example 5-1

[0164] This example discloses the synthesis of compound b9-1-5-BD-WJ (i.e., in compound b9, a = b = e = 0, d = 1, X = CH3, c = 5, R1= WJ), the reaction scheme of which is:

[0165]

[0166] To 30 ul of hydrazine hydrate dissolved in anhydrous dichloromethane (1.5 mL) was added b8-1-5-BD-WJ (100 mg, 0.082 mmol) and stirred at room temperature for 1 hour. After completion, the reaction was poured into ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine three times, collected and dried over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, the organic solvent was removed by rotary evaporation to give 65 mg of crude product b9-1-5-BD-WJ as a colorless transparent oil, which was used directly in the next step.

[0167] Example 5-2

[0168] This example discloses the synthesis of compound b9-1-5-BID-WJ (i.e., in compound b9, a = b = e = 0, d = 1, X = I, c = 5, R1= WJ), the reaction scheme of which is:

[0169]

[0170] To 30 ul of hydrazine hydrate dissolved in anhydrous dichloromethane (1.5 mL) was added b8-1-5-BID-WJ (110 mg, 0.082 mmol) and stirred at room temperature for 1 hour. After completion, the reaction was poured into ice water and extracted with ethyl acetate. The organic phase was washed with saturated brine three times, collected and dried over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, the organic solvent was removed by rotary evaporation to give 58.5 mg of crude product b9-1-5-BID-WJ as a colorless transparent oil, which was used directly in the next step.

[0171] Example 5-3

[0172] This example discloses the synthesis of compound b9-2-4-PN-WJ (i.e., in compound b9, a = e = 1, c = d = 0, b = 4, R1= WJ), the reaction scheme of which is:

[0173]

[0174] Dissolve 30 ul hydrazine hydrate in anhydrous dichloromethane (1.5 mL) and stir well, then add b8-2-4-BD-WJ (115.5 mg, 0.082 mmol) and stir at room temperature for 1 hour. After completion, pour the reaction mixture into ice water and extract with ethyl acetate. Wash the organic phase with saturated brine three times, collect the organic phase and dry over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, remove the organic solvent by rotary evaporation to obtain 56.8 mg of crude product b9-2-4-BD-WJ as colorless transparent oil. The crude product is used directly in the next step.

[0175] Example 5-4

[0176] This example discloses the synthesis of compound b9-2-4-BD-WJ (i.e., in compound b9, a = d = 1, X = CH3, b = 4, c = e = 0, R1= WJ), whose reaction formula is:

[0177]

[0178] Dissolve 30 ul hydrazine hydrate in anhydrous dichloromethane (1.5 mL) and stir well, then add b8-2-4-BD-WJ (115.5 mg, 0.082 mmol) and stir at room temperature for 1 hour. After completion, pour the reaction mixture into ice water and extract with ethyl acetate. Wash the organic phase with saturated brine three times, collect the organic phase and dry over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, remove the organic solvent by rotary evaporation to obtain 56.8 mg of crude product b9-2-4-BD-WJ as colorless transparent oil. The crude product is used directly in the next step.

[0179] Example 6-1

[0180] This example discloses the synthesis of compound b11-1-5-BD-WJ-DOTA (i.e., in compound b11, a = b = e = 0, d = 1, X = CH3, c = 5, R1= WJ, R2= DOTA), whose reaction formula is:

[0181]

[0182] Dissolve b10-DOTA (45 mg, 0.096 mmol) in acetonitrile (2 mL), then add NHS (13.2 mg, 0.115 mmol), HBTU (41.8 mg, 0.110 mmol) and diisopropylethylamine (19 mg, 0.147 mmol), and stir at room temperature for 30 minutes. Then add b9-1-5-BD-WJ (77.4 mg, 0.074 mmol) and stir at room temperature for 10 hours. Pour the reaction mixture into ice water and extract with dichloromethane. Wash the organic phase with saturated brine three times, collect the organic phase and dry over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, remove the organic solvent by rotary evaporation. The product is used directly in the next step.

[0183] Example 6-2

[0184] This example discloses the synthesis of compound b11-1-5-BID-WJ-DOTA (i.e., in compound b11, a = b = e = 0, d = 1, X = I, c = 5, R1= WJ, R2= DOTA), the reaction scheme of which is:

[0185]

[0186] b10-DOTA (45 mg, 0.096 mmol) was dissolved in acetonitrile (2 mL), followed by the addition of NHS (13.2 mg, 0.115 mmol), HBTU (41.8 mg, 0.110 mmol) and diisopropylethylamine (19 mg, 0.147 mmol). After stirring at room temperature for 30 minutes, b9-1-5-BID-WJ (87 mg, 0.074 mmol) was added, and the reaction was stirred at room temperature for 10 hours. The reaction was poured into ice water, extracted with dichloromethane, washed with saturated brine three times, and the organic phase was collected and dried over anhydrous sodium sulfate overnight. After the drying agent was removed by filtration, the organic solvent was removed by rotary evaporation, and the product was used directly in the next reaction.

[0187] Example 6-3

[0188] This example discloses the synthesis of compound b11-1-5-BD-WJ-DOTAGA (i.e., in compound b11, a = b = e = 0, d = 1, c = 5, R1= WJ, R2= DOTAGA), the reaction scheme of which is:

[0189]

[0190] b10-DOTAGA (67.3 mg, 0.096 mmol) was dissolved in acetonitrile (2 mL), followed by the addition of NHS (13.2 mg, 0.115 mmol), HBTU (41.8 mg, 0.110 mmol) and diisopropylethylamine (19 mg, 0.147 mmol). After stirring at room temperature for 30 minutes, b9-1-5-BD-WJ (78 mg, 0.074 mmol) was added, and the reaction was stirred at room temperature for 10 hours. The reaction was poured into ice water, extracted with dichloromethane, washed with saturated brine three times, and the organic phase was collected and dried over anhydrous sodium sulfate overnight. After the drying agent was removed by filtration, the organic solvent was removed by rotary evaporation, and the product was used directly in the next reaction.

[0191] Example 6-4

[0192] This example discloses the synthesis of compound bll-2-4-PN-WJ-DOTA (i.e., in compound bll, a = e = 1, c = d = 0, b = 4, R1= WJ, R2= DOTA), whose reaction formula is:

[0193]

[0194] b10-DOTA (45 mg, 0.096 mmol) was dissolved in acetonitrile (2 mL), and then NHS (13.2 mg, 0.115 mmol), HBTU (41.8 mg, 0.110 mmol) and diisopropylethylamine (19 mg, 0.147 mmol) were added. After stirring at room temperature for 30 minutes, b9-2-4-PN-WJ (85.5 mg, 0.074 mmol) was added, and stirring was performed at room temperature for 10 hours. The reaction solution was poured into ice water, extracted with dichloromethane, washed with saturated brine for 3 times, and then the organic phase was collected and dried over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, the organic solvent was removed by rotary evaporation, and the product was directly used in the next reaction.

[0195] Example 6-5

[0196] This example discloses the synthesis of compound bll-2-4-PN-WJ-DOTA (i.e., in compound bll, a = e = 1, c = d = 0, b = 4, R1= WJ, R2= DOTA), whose reaction formula is:

[0197]

[0198] b10-DOTA (45 mg, 0.096 mmol) was dissolved in acetonitrile (2 mL), and then NHS (13.2 mg, 0.115 mmol), HBTU (41.8 mg, 0.110 mmol) and diisopropylethylamine (19 mg, 0.147 mmol) were added. After stirring at room temperature for 30 minutes, b9-2-4-PN-WJ (85.5 mg, 0.074 mmol) was added, and stirring was performed at room temperature for 10 hours. The reaction solution was poured into ice water, extracted with dichloromethane, washed with saturated brine for 3 times, and then the organic phase was collected and dried over anhydrous sodium sulfate overnight. After removing the drying agent by filtration, the organic solvent was removed by rotary evaporation, and the product was directly used in the next reaction.

[0199] Example 7-1

[0200] This example discloses the synthesis of compound bll-2-4-PN-WJ-DOTA (i.e., in compound bll, a = e = 1, c = d = 0, b = 4, R1= WJ, R2= DOTA), whose reaction formula is:

[0201]

[0202] b11-1-5-BD-WJ-DOTA (25.8 mg, 0.016 mmol) was dissolved in a mixed solvent of dichloromethane (0.5 ml) and trifluoroacetic acid (0.5 ml), and the reaction solution was stirred at room temperature for 12 hours. The organic solvent and trifluoroacetic acid were removed by rotary evaporation under reduced pressure, the product was dissolved in methanol again, and the crude product was obtained by extraction with diethyl ether, and then the product of formula I-1-5-BD-WJ-DOTA was obtained by separation and purification with preparative high performance liquid chromatography, with a total of 3.4 mg and a yield of 16.7%.

[0203] The compound of formula I-1-5-BD-WJ-DOTA was subjected to LC-MS analysis, and the mass spectrum is shown in FIG. 1. Figure 1

[0204] The mass spectrometer was Agilent 1200 series 6120 model, electrospray protonation (ESI), and the HPLC conditions were as follows: Waters XBridge C18 column (150 mm x 4.6 mm x 3.5 μm), flow rate: 1.0 ml / min, column temperature: 40°C, gradient: acetonitrile (0.05% TFA): water (0.05%), in which the acetonitrile was increased from 5% to 100% in 10 minutes, and eluted at 100% for 10 minutes.

[0205] Example 7-2

[0206] This example discloses the synthesis of the compound of formula I-1-5-BID-WJ-DOTA (i.e., in the compound of formula I, a = b = e = 0, d = 1, X = I, c = 5, R1 = WJ, R2 = DOTA), and the reaction formula is as follows:

[0207]

[0208] b11-1-5-BID-WJ-DOTA (27.5 mg, 0.016 mmol) was dissolved in a mixed solvent of dichloromethane (0.5 ml) and trifluoroacetic acid (0.5 ml), and the reaction solution was stirred at room temperature for 12 hours. The organic solvent and trifluoroacetic acid were removed by rotary evaporation under reduced pressure, the product was dissolved in methanol again, and the crude product was obtained by extraction with diethyl ether, and then the product of formula I-1-5-BID-WJ-DOTA was obtained by separation and purification with preparative high performance liquid chromatography, with a total of 4.2 mg and a yield of 19%.

[0209] Example 7-3

[0210] This example discloses the synthesis of the compound of formula I-1-5-BD-WJ-DOTAGA (i.e., in the compound of formula I, a = b = e = 0, d = 1, c = 5, R1 = WJ, R2 = DOTAGA), and the reaction formula is as follows:​

[0211]

[0212] b11-1-5-BD-WJ-DOTAGA (27.8 mg, 0.016 mmol) was dissolved in a mixture of dichloromethane (0.5 ml) and trifluoroacetic acid (0.5 ml), and the reaction was stirred at room temperature for 12 hours. The organic solvent and trifluoroacetic acid were removed by rotary evaporation under reduced pressure, the product was dissolved in methanol again, and the crude product was obtained by extraction with diethyl ether, and then purified by preparative high performance liquid chromatography to obtain the product of formula I-1-5-BD-WJ-DOTAGA, a total of 4.9 mg, yield: 22.8%.

[0213] Example 7-4

[0214] This example discloses the synthesis of the compound of formula I-2-4-BD-WJ-DOTA (i.e., the compound of formula I, a = d = 1, X = CH3, b = 4, c = e = 0, R1 = WJ, R2 = DOTA), the reaction formula of which is as follows:

[0215]

[0216] b11-2-4-BD-WJ-DOTA (28.8 mg, 0.016 mmol) was dissolved in a mixture of dichloromethane (0.5 ml) and trifluoroacetic acid (0.5 ml), and the reaction was stirred at room temperature for 12 hours. The organic solvent and trifluoroacetic acid were removed by rotary evaporation under reduced pressure, the product was dissolved in methanol again, and the crude product was obtained by extraction with diethyl ether, and then purified by preparative high performance liquid chromatography to obtain the product of formula I-2-4-BD-WJ-DOTA, a total of 4.4 mg, yield: 18.8%.

[0217] The compound of formula I-2-4-BD-WJ-DOTA was subjected to LC-MS analysis, and the mass spectrum is shown in FIG. 2. Figure 2

[0218] The mass spectrometer was Agilent 1200 series 6120 model, electrospray protonation (ESI), and the HPLC conditions were as follows: Waters XBridge C18 column (150 mm x 4.6 mm x 3.5 μm), flow rate: 1.0 ml / min, column temperature: 40 °C, gradient: acetonitrile (0.05% TFA): water (0.05%), in which the acetonitrile was increased from 5% to 100% in 10 minutes, and eluted at 100% for 10 minutes.

[0219] Example 7-5

[0220] ​This example discloses the synthesis of compound Formula I-2-4-PN-WJ-DOTA (i.e. in Formula I, a = e = 1, c = d = 0, b = 4, R1= WJ, R2= DOTA), whose reaction formula is as follows:

[0221]

[0222] b11-2-4-PN-WJ-DOTA (27.5 mg, 0.016 mmol) was dissolved in a mixed solvent of dichloromethane (0.5 ml) and trifluoroacetic acid (0.5 ml), and the reaction solution was stirred at room temperature for 12 hours. The organic solvent and trifluoroacetic acid were removed by rotary evaporation under reduced pressure, the product was dissolved in methanol again, and the crude product was obtained by extraction with diethyl ether, and then the product Formula I-2-4-PN-WJ-DOTA was obtained by preparative high performance liquid chromatography separation and purification, a total of 5.5 mg, yield: 22.3%.

[0223] Formula I-2-4-PN-WJ-DOTA was subjected to LC-MS analysis, and its mass spectrum is shown in Figure Figure 3

[0224] The mass spectrometer was Agilent 1200 series 6120 model, electrospray protonation (ESI), and the HPLC conditions were as follows: Waters XBridge C18 column (150 mm x 4.6 mm x 3.5 μm), flow rate: 1.0 ml / min, column temperature: 40 °C, gradient: acetonitrile (0.05% TFA): water (0.05%), in which the acetonitrile was increased from 5% to 100% in 10 minutes, and eluted at 100% for 10 minutes.

[0225] Example 8

[0226] This example discloses the synthesis of compound Formula I-2-4-PN-WJ-DOTA (i.e. in Formula I, a = e = 1, c = d = 0, b = 4, R1= WJ, R2= DOTA), whose reaction formula is as follows: 68 Ga-labeled compound 68 Ga-Formula II-1-5-BD-WJ-DOTA, 68 Ga-Formula II-1-5-BD-WJ-DOTAGA, 68 Ga-Formula II-1-5-BID-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 Ga-Formula II-2-4-PN-WJ-DOTA, whose reaction formula is as follows:

[0227]

[0228]

[0229] ​The specific method is as follows: NaAc / HAc buffer solution (pH=4.2, 1 mL) is mixed with 0.85% normal saline (1 mL) at room temperature, then 10 μL (10 μg) of formula I-5-BD-WJ-DOTA is added, and after mixing, 0.85% normal saline (1 mL) is added, and the mixture is mixed and reacted at room temperature for 30 minutes. The mixture is then passed through a C18lighting reverse-phase column, washed with normal saline, and collected as waste liquid. The reverse-phase column is then washed with 50% medical alcohol (1 mL), washed with 0.85% normal saline (5 mL), and the alcohol washing liquid and normal saline washing liquid are collected. The retention time and radiochemical purity of the product are measured by high performance liquid chromatography (acetonitrile / water, acetonitrile 10% to 90% in 15 minutes, both water and acetonitrile containing 0.1% trifluoroacetic acid). The radioactive peak retention time of the product is 9.688 min, and the labeling rate is 98.95%. 68 GaCl3high-purity hydrochloric acid solution (5 mCi, 2 mL, 0.05 mol / L) is heated to 90°C for 15 minutes, passed through a C18lighting reverse-phase column, washed with normal saline, and collected as waste liquid. The reverse-phase column is then washed with 50% medical alcohol (1 mL), washed with 0.85% normal saline (5 mL), and the alcohol washing liquid and normal saline washing liquid are collected. The retention time and radiochemical purity of the product are measured by high performance liquid chromatography (acetonitrile / water, acetonitrile 10% to 90% in 15 minutes, both water and acetonitrile containing 0.1% trifluoroacetic acid). The radioactive peak retention time of the product is 9.688 min, and the labeling rate is 98.95%.

[0230] 68 Ga-Formula II-1-5-BD-WJ-DOTAGA, 68 Ga-Formula II-1-5-BID-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 Ga-Formula II-2-4-PN-WJ-DOTA are prepared by the same method as Ga-Formula II-1-5-BD-WJ-DOTA. 68 68 Ga-Formula II-1-5-BD-WJ-DOTAGA has a labeling rate of 99.32%, 68 Ga-Formula II-1-5-BID-WJ-DOTA has a labeling rate of 98.98%, 68 Ga-Formula II-2-4-BD-WJ-DOTA has a labeling rate of 98.74%, 68 Ga-Formula II-2-4-PN-WJ-DOTA has a labeling rate of 99.11%.

[0231] Formula I-1-5-BD-WJ-DOTA, Formula I-1-5-BID-WJ-DOTA, Formula I-1-5-BD-WJ-DOTAGA, Formula I-2-4-PN-WJ-DOTA, and Formula I-2-4-BD-WJ-DOTA in this example are prepared by the methods of Examples 7-1, 7-2, 7-3, 7-4, and 7-5, respectively.

[0232] Formula 68 Ga-Formula II-1-5-BD-WJ-DOTA is shown in FIG. 1. The radio high performance liquid chromatogram of Ga-Formula II-1-5-BID-WJ-DOTA is shown in FIG. 2. The radio high performance liquid chromatogram of Ga-Formula II-2-4-BD-WJ-DOTA is shown in FIG. 3. The radio high performance liquid chromatogram of Ga-Formula II-2-4-PN-WJ-DOTA is shown in FIG. 4. Figure 4 Formula 68 ​The radioactive high-performance liquid chromatogram of Ga-type II-2-4-BD-WJ-DOTA is attached. Figure 5 As shown.

[0233] The formula obtained in this embodiment 68 The radioactive high-performance liquid chromatogram of Ga-type II-2-4-PN-WJ-DOTA is attached. Figure 6 As shown.

[0234] HPLC conditions were as follows: Angilent C18 column (250 mm x 4.6 mm x 3.5 μm), flow rate: 1.0 mL / min, column temperature: room temperature. The gradient was acetonitrile (0.1% TFA): water (0.1% TFA), wherein the acetonitrile was increased from 10% to 90% over 15 minutes, and isocratic elution was performed at 90% for 10 minutes.

[0235] Example 9

[0236] This embodiment discloses 177 LuCl3-labeled compound of formula II 177 Lu-style II-1-5-BD-WJ-DOTA 177 Lu-style

[0237] Ⅱ-1-5-BD-WJ-DOTAGA 177 Lu-style II-1-5-BID-WJ-DOTA, 177 Lu-style II-2-4-BD-WJ-DOTA, 177 Lu-style

[0238] The preparation of Ⅱ-2-4-PN-WJ-DOTA is carried out by the following reaction:

[0239]

[0240]

[0241] Specifically, the following steps were taken: NaAc / HAc buffer solution (pH = 4.6, 1 mL) was mixed with 0.85% physiological saline (1 mL) at room temperature, then the precursor I-1-5-BD-WJ-DOTA (20 μL, 20 μg) was added, mixed thoroughly, and then... 177A high-purity hydrochloric acid solution of LuCl3 (5 μL, 0.04 mol / L high-purity hydrochloric acid, specific activity 1 mCi / μL) was heated to 90℃ and reacted for 15 minutes. The solution was then passed through a C18 Lighting reversed-phase column, washed with saline, and collected as waste liquid. The reversed-phase column was then washed with 50% medical alcohol (0.2 mL) and 0.85% physiological saline (2 mL). The alcohol solution and washing liquid were collected, and their high-performance liquid chromatography (HPLC) was measured (acetonitrile / water, acetonitrile 10% to 90% within 15 minutes, water and acetonitrile both containing 0.1% trifluoroacetic acid). The retention time of the radioactive peak of the product was 7.69 min, and the labeling rate was 98.29%.

[0242] 177 Lu-style II-1-5-BD-WJ-DOTAGA, 177 Lu-style II-1-5-BID-WJ-DOTA, 177 Lu-style II-2-4-BD-WJ-DOTA 177 Preparation method of Lu-type II-2-4-PN-WJ-DOTA and 177 Lu-style II-1-5-BD-WJ-DOTA is the same. 177 The marking rate of Lu--Form II-1-5-BD-WJ-DOTAGA was 99.13%. 177 The labeling rate of Lu-type II-1-5-BID-WJ-DOTA was 98.74%. 177 The labeling rate of Lu-type II-2-4-BD-WJ-DOTA was 97.94%. 177 The labeling rate of Lu-type II-2-4-PN-WJ-DOTA was 99.09%.

[0243] The formulas I-1-5-BD-WJ-DOTA, I-1-5-BID-WJ-DOTA, I-1-5-BD-WJ-DOTAGA, I-2-4-PN-WJ-DOTA, and I-2-4-BD-WJ-DOTA used in this embodiment were prepared according to the methods of Examples 7-1, 7-2, 7-3, 7-4, and 7-5, respectively. The formulas prepared in this embodiment... 177 The radioactive high-performance liquid chromatogram of Lu-type II-1-5-BD-WJ-DOTA is attached. Figure 7 As shown. The formula obtained in this embodiment 177 The radioactive high-performance liquid chromatogram of Lu-type II-2-4-BD-WJ-DOTA is attached. Figure 8 As shown.

[0244] The formula obtained in this embodiment 177 The radioactive high-performance liquid chromatogram of Lu-form II-2-4-PN-WJ-DOTA is attached. Figure 9As shown. HPLC conditions were: Angilent C18 column (250 mm x 4.6 mm x 3.5 μm), flow rate: 1.0 mL / min, column temperature: room temperature. The gradient was acetonitrile (0.1% TFA): water (0.1% TFA), wherein the acetonitrile was increased from 10% to 90% over 15 minutes, and isocratic elution was performed at 90% for 10 minutes.

[0245] Example 10

[0246] This embodiment discloses 225 Ac-labeled compounds of formula II 225 Ac-Type II-1-5-BD-WJ-DOTA, 225 Ac-type

[0247] Ⅱ-1-5-BD-WJ-DOTAGA、 225 The preparation of Ac-form II-1-5-BID-WJ-DOTA is carried out by the following reaction:

[0248]

[0249] Add 0.1M Tris buffer (pH 9.0, 1.0 mL) to a 5 mL EP tube, then add... 225 The reaction mixture consisted of 0.1 mL of Ac-hydrochloric acid solution (0.3 MBq) and 20 μL (20 μg) of formula I-1-5-BD-WJ-DOTA. After preheating to 85°C in a metal bath, the reaction system was placed in the metal heating bath and heated at 85°C for 5 minutes. Heating was then stopped, and 2 mL of sterile water for injection was added to the reaction solution. The mixture was passed through a C18 micro-separation column, and the column was washed with 5 mL of sterile water for injection, collecting the waste liquid. The C18 column was then rinsed with 0.5 mL of 50% medical alcohol, followed by 2 mL of sterile water for injection. The alcohol washing solution and 2 mL of sterile water for injection were collected as the product. 10 μL of the product solution was extracted and analyzed by thin-layer chromatography (TLC) for radiochemical purity; the labeling rate was 98.55%.

[0250] 225 Ac-Type II-1-5-BD-WJ-DOTAGA 225 Preparation method of Ac-type II-1-5-BID-WJ-DOTA and 225 The Ac-type II-1-5-BD-WJ-DOTA is the same. 225 The Ac-type II-1-5-BD-WJ-DOTAGA labeling rate was 98.79%. 225 The Ac-type II-1-5-BID-WJ-DOTA labeling rate was 99.02%.

[0251] The formula I-1-5-BD-WJ-DOTA, formula I-1-5-BID-WJ-DOTA and formula I-1-5-BD-WJ-DOTAGA in this example are prepared according to the methods of example 7-1, 7-2 and 7-3 respectively.

[0252] Example 11

[0253] This example discloses 64 Cu-labeled formula II compound 64 Cu-formula II-1-5-BD-WJ-DOTA, 64 Cu-formula II-1-5-BD-WJ-DOTAGA, 64 The preparation of Cu-formula II-1-5-BID-WJ-DOTA is shown in the following reaction formula:

[0254]

[0255]

[0256] The specific steps are as follows: in a 5 mL EP tube, add 1.0 M NaAc / HAc buffer (pH 4.4, 1.0 mL), then add the purified 64 CuCl2solution (1 mL, 3.8 MBq) and formula I-1-5-BD-WJ-DOTA (20 μL, 20 μg). After preheating to 85°C in a metal bath, place the above reaction system into the metal heating bath, heat at 85°C for 10 minutes, stop heating, and add 2 mL of sterile water for injection to the reaction solution. Pass through a C18 microcolumn and wash the column with 5 mL of sterile water for injection, and collect the waste liquid. The C18 column is then eluted with 0.5 mL of 50% medical alcohol, and then washed with 2 mL of sterile water for injection. The alcohol eluate and 2 mL of sterile water for injection are collected as the product, and 10 μL of the product solution is taken for radiochemical purity determination by radio-HPLC. The labeling yield is 97.56%.

[0257] 64 Cu-formula II-1-5-BD-WJ-DOTAGA, 64 The preparation method of Cu-formula II-1-5-BID-WJ-DOTA is the same as 64 Cu-formula II-1-5-BD-WJ-DOTA, 64 The labeling yield of Cu-formula II-1-5-BD-WJ-DOTAGA is 98.99%, 64 The labeling yield of Cu-formula II-1-5-BID-WJ-DOTA is 98.61%.

[0258] The formula I-1-5-BD-WJ-DOTA, formula I-1-5-BID-WJ-DOTA, formula I-1-5-BD-WJ-DOTAGA used in this example were prepared according to the methods of example 7-1, 7-2, 7-3 respectively. Test Example 1

[0259] This test example discloses 68 Ga-labeled formula II compound 68 Ga-formula II-1-5-BD-WJ-DOTA, 68 Ga-formula II-2-4-BD-WJ-DOTA, 68 Ga-formula II-2-4-PN-WJ-DOTA PBS stability experiment, specifically:

[0260] 20 μL of the label was taken in parallel 68 Ga-formula II-1-5-BD-WJ-DOTA was dissolved in 3 180 μL PBS buffer and incubated at 37°C for 30 min, 60 min, and 120 min, and sampled at each time point, and the change in sample radio peak retention time was determined by high performance liquid chromatography.

[0261] The same method was used to determine 68 Ga-formula II-2-4-BD-WJ-DOTA, 68 Ga-formula II-2-4-PN-WJ-DOTA in PBS.

[0262] Test results: sample 68 Ga-formula II-1-5-BD-WJ-DOTA was dissolved in 3 180 μL PBS buffer and incubated at 37°C for 30 min, 60 min, and 120 min, and sampled at each time point, and the change in sample radio peak retention time was determined by high performance liquid chromatography. 68 Ga-formula II-2-4-BD-WJ-DOTA was dissolved in 3 180 μL PBS buffer and incubated at 37°C for 30 min, 60 min, and 120 min, and sampled at each time point, and the change in sample radio peak retention time was determined by high performance liquid chromatography. 68 Ga-formula II-2-4-PN-WJ-DOTA was dissolved in 3 180 μL PBS buffer and incubated at 37°C for 30 min, 60 min, and 120 min, and sampled at each time point, and the change in sample radio peak retention time was determined by high performance liquid chromatography. 68 Ga-formula II-1-5-BD-WJ-DOTA, 68 Ga-formula II-2-4-BD-WJ-DOTA, 68 Ga-formula II-2-4-PN-WJ-DOTA label has good stability in PBS.

[0263] Table 1 68 Ga-formula II-1-5-BD-WJ-DOTA, 68 Ga-formula II-2-4-BD-WJ-DOTA, 68PBS stability test results of Ga-type II-2-4-PN-WJ-DOTA

[0264] Time 30 min 60 min 120 min 68 Radiochemical purity of Ga-Formula II-1-5-BD-WJ-DOTA ​ 99.7％±1.2％ 99.35％±0.9％ 99.23％±1.4％ 68 Radiochemical purity of Ga-Formula II-2-4-BD-WJ-DOTA ​ 99.58％±0.69％ 99.23％±0.58％ 98.51％±0.19％ 68 Radiochemical purity of Ga-Formula II-2-4-PN-WJ-DOTA ​ 99.47％±0.15％ 98.57％±0.14％ 97.70％±0.98％

[0265] Experimental Example 2

[0266] This experimental example discloses 68 Ga-labeled compound of formula II 68 Ga-Formula II-1-5-BD-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 The fetal bovine serum stability assay for Ga-type II-2-4-PN-WJ-DOTA is as follows:

[0267] Take 200 μL of fetal bovine serum in parallel into three 2 mL EP tubes, then add radiochemically pure (>99%)... 68 100 μL of Ga-type II-1-5-BD-WJ-DOTA marker (specific activity: 0.5 μCi / μL) was incubated at 37 °C for 30 min, 60 min, and 120 min. At each time point, an equal volume of acetonitrile to serum was added to the sample, the precipitate was shaken, centrifuged, and 20 μL of the supernatant was collected. The radiochemical purity of the sample was determined by radioactive high-performance liquid chromatography.

[0268] Same method determination 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 Stability of Ga-type II-2-4-PN-WJ-DOTA in fetal bovine serum.

[0269] Test results: Sample 68 The Ga-type II-1-5-BD-WJ-DOTA sample had a radiochemical purity of 98.16% after 30 minutes and maintained a radiochemical purity of 97.50% after 2 hours. 68 The radiochemical purity of Ga-type II-2-4-BD-WJ-DOTA was 97.87% after 30 minutes. 68 The radiochemical purity of Ga-type II-2-4-PN-WJ-DOTA was 98.59% after 30 minutes; after 2 hours, both maintained high radiochemical purity, at 96.48% and 93.07%, respectively. Therefore, 68 Ga-Formula II-1-5-BD-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 The Ga-type II-2-4-PN-WJ-DOTA marker exhibits good stability in fetal bovine serum.

[0270] The radio-HPLC profile of the stability (120 min) of the compound in fetal bovine serum is shown in Figure 1. Figure 10 , 11 and 12.

[0271] Table 2 68 Ga-Formula II-1-5-BD-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 The results of the fetal bovine serum stability experiment of Ga-Formula II-2-4-PN-WJ-DOTA

[0272] Time 30 min 60 min 120 min 68 Radiochemical purity of Ga-Formula II-1-5-BD-WJ-DOTA ​ 98.16％±0.7％ 98.43％±0.9％ 97.50％±1.2％ 68 Radiochemical purity of Ga-Formula II-2-4-BD-WJ-DOTA ​ 97.87％±1.47％ 96.86％±1.18％ 96.48％±1.06％ 68 Radiochemical purity of Ga-Formula II-2-4-PN-WJ-DOTA ​ 98.59％±0.45％ 96.77％±1.47％ 93.07％±1.73％

[0273] Test Example 3

[0274] This test example discloses 225 Ac-labeled Formula II compound 225 The PBS stability experiment of Ac-Formula II-1-5-BD-WJ-DOTA, specifically:

[0275] 20 μL of the labeled Formula II compound was taken in parallel 225 Ac-Formula II-1-5-BD-WJ-DOTA was dissolved in 3 80 μL PBS buffers, and incubated at 37°C for 30 min, 60 min, and 120 min, and sampled at each time point, and the change in the retention time of the radio peak of the sample was determined by thin layer chromatography.

[0276] Test results: sample 225 The radiochemical purity of Ac-Formula II-1-5-BD-WJ-DOTA was 98.50% at 30 minutes, and remained 98.01% after 2 hours. Therefore, 225 Ac-Formula II-1-5-BD-WJ-DOTA is stable in PBS.

[0277] Table 3 225 The results of the PBS stability experiment of Ac-Formula II-1-5-BD-WJ-DOTA

[0278] Time 30 min 60 min 120 min 225 Radiochemical purity of Ac-Formula II-1-5-BD-WJ-DOTA ​ 98.50％±0.8％ 99.20％±1.4％ 98.01％±0.9％

[0279] Test Example 4

[0280] This test example discloses 225 Ac-labeled Formula II compound 225 The fetal bovine serum stability experiment of Ac-Formula II-1-5-BD-WJ-DOTA, specifically:

[0281] 100 μL of fetal bovine serum was taken in parallel in 2 mL of 3 EP tubes, and then the labeled compound with a radiochemical purity of >99% was added225 Ac-Formula II-1-5-BD-WJ-DOTA 100 μL, incubated at 37°C for 30 min, 60 min, 120 min. At each time point, an equal amount of acetonitrile as the serum was added to the sample, and the sample was shaken to precipitate, and after centrifugation, the supernatant was spotted on a thin layer chromatography paper, and the radiochemical purity of the sample was detected by thin layer chromatography.

[0282] Test results: sample 225 The radiochemical purity of Ac-Formula II-1-5-BD-WJ-DOTA was 98.17% at 30 min, and remained 95.39% after 2 hours. Therefore, 225 The Cu-Formula II-1-5-BD-WJ-DOTA label has good stability in the fetal bovine serum.

[0283] Table 4 225 The results of the fetal bovine serum stability experiment of Ac-Formula II-1-5-BD-WJ-DOTA

[0284] Time 30 min 60 min 120 min 225 Radiochemical purity of Ac-Formula II-1-5-BD-WJ-DOTA ​ 98.17％±0.6％ 98.09％±0.3％ 95.39％±0.8％

[0285] Test Example 5

[0286] This test example discloses 64 Cu-labeled Formula II compound 64 The PBS stability experiment of Cu-Formula II-1-5-BD-WJ-DOTA is as follows:

[0287] 20 μL of the label was taken in parallel 64 Cu-Formula II-1-5-BD-WJ-DOTA was dissolved in 3 PBS buffers of 180 μL, and incubated at 37°C for 30 min, 60 min, 120 min, and sampled at each time point, and the change in the retention time of the radiochemical peak of the sample was determined by high performance liquid chromatography.

[0288] The test results: the radiochemical purity of the sample was 98.7% at 30 min, and remained 97.62% after 2 hours. Therefore, 64 The Cu-Formula II-1-5-BD-WJ-DOTA label has good stability in the PBS.

[0289] Table 5 64 The results of the PBS stability experiment of Cu-Formula II-1-5-BD-WJ-DOTA

[0290] Time 30 min 60 min 120 min 64 Radiochemical purity of Cu-formula II-1-5-BD-WJ-DOTA ​ 98.7％±0.5％ 98.1％±1.1％ 97.62％±1.6％

[0291] Test Example 6

[0292] This test example discloses 64 Cu-labeled Formula II compound64 The fetal bovine serum stability experiment of Cu-Formula II-1-5-BD-WJ-DOTA is specifically as follows:

[0293] 200 μL of fetal bovine serum was taken in parallel in 2 mL of 3 EP tubes, and 100 μL of radiolabeled compound with a radiochemical purity of >99% was added. Incubation was performed at 37°C for 30 min, 60 min and 120 min. At each time point, an equal amount of acetonitrile was added to the sample, and the sample was shaken and precipitated. After centrifugation, 20 μL of supernatant was taken, and the radiochemical purity of the sample was detected by radio-HPLC.

[0294] Test results: The radiochemical purity of the sample was 98.5% at 30 min, and remained at 96.7% after 2 h. Therefore, 64 The Cu-Formula II-1-5-BD-WJ-DOTA labeled compound has good stability in fetal bovine serum.

[0295] Table 6 64 The fetal bovine serum stability experiment results of Cu-Formula II-1-5-BD-WJ-DOTA

[0296] Time Time Figure 13 Figure 14 64 Radiochemical purity of Cu-formula II-1-5-BD-WJ-DOTA ​ 98.5％±1.9％ 97.9％±1.3％ 96.7％±2.1％

[0297] Test Example 7

[0298] This test example discloses 177 Lu-labeled Formula II compound 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 The PBS stability experiment of Lu-Formula II-2-4-PN-WJ-DOTA is specifically as follows:

[0299] 20 μL of labeled compound was taken 177 Lu-Formula II-1-5-BD-WJ-DOTA was dissolved in 180 μL of PBS buffer and incubated at 37°C for 1 h, 4 h and 24 h, and sampled at each time point. The radiochemical purity of the sample was determined by HPLC.

[0300] The radiochemical purity of the sample was determined by HPLC. 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-PN-WJ-DOTA in PBS.

[0301] Test results: The radiochemical purity of the sample 177 The radiochemical purity of Lu-Formula II-1-5-BD-WJ-DOTA was 98.50% at 1 h, and remained at 96.54% after 24 h. The radiochemical purity of the sample 177The radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA was 98.84% at 1h, and the sample 177 The radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA was 98.84% at 1h, and the sample 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA was stable in PBS.

[0302] Table 7 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 The PBS stability experiment results of Lu-Formula II-2-4-PN-WJ-DOTA

[0303] Figure 15 1h 4h 24h 177 Radiochemical purity of Lu-formula II-1-5-BD-WJ-DOTA ​ 98.50％±1.5％ 98.29％±1.1％ 96.54％±0.6％ 177 Radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA ​ 98.84％±1.2％ 97.79％±2.19％ 96.17％±1.24％ 177 Radiochemical purity of Lu-Formula II-2-4-PN-WJ-DOTA ​ 98.82％±1.87％ 98.47％±3.61％ 97.9％±0.51％

[0304] Test Example 8

[0305] This test example discloses 177 Lu-labeled Formula II compound 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 The fetal bovine serum stability experiment of Lu-Formula II-2-4-PN-WJ-DOTA, specifically:

[0306] 100 μL of fetal bovine serum was taken in parallel in 2 mL of 3 EP tubes, and 100 μL of the label with a radiochemical purity of >99% was added, and incubated at 37°C for 1h, 4h, and 24h. At each time point, an equal amount of acetonitrile was added to the sample, and the precipitate was shaken and centrifuged, and 20 μL of the supernatant was taken and detected by radio-high performance liquid chromatography to detect the radiochemical purity of the sample.

[0307] The same method was used to determine 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 The stability of Lu-Formula II-2-4-PN-WJ-DOTA in fetal bovine serum.

[0308] Test results: sample 177 The radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA was 98.84% at 1h, and the sample 177The radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA was 96.4% at 1h, and the sample 177 The radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA was 96.4% at 1h, and the sample 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA,

[0309] Table 8 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 The results of the stability experiment of Lu-Formula II-2-4-BD-WJ-DOTA in fetal bovine serum

[0310] Figure 16 1h 4h 24h 177 Radiochemical purity of Lu-Formula II-1-5-BD-WJ-DOTA ​ 92.68％±1.2％ 90.06％±0.9％ 88.15％±1.2％ 177 Radiochemical purity of Lu-Formula II-2-4-BD-WJ-DOTA ​ 96.4％±2.03％ 94.5％±1.38％ 90.85％±2.03％ 177 Radiochemical purity of Lu-Formula II-2-4-PN-WJ-DOTA ​ 98.7％±1.07％ 97.9％±1.56％ 94.78％±0.82％

[0311] Test Example 9

[0312] This test example discloses 68 Ga-Formula II-2-4-BD-WJ-DOTA, 68 Ga-Formula II-2-4-BD-WJ-DOTA, and 68 Ga-Formula II-2-4-BD-WJ-DOTA, and

[0313] LNCaP cells were implanted in the left anterior limb of NOD / SCID mice under the armpit, and the animals were bred in a SPF level animal room for about 4 weeks, and the tumor mass was about 0.8 cm in diameter. About 100 μL of high radiochemical purity drug (radiochemical purity > 98.5%) with a dose of about 50 μCi was injected through the tail vein. Imaging was performed at 10 minutes, 30 minutes, 60 minutes and 120 minutes, respectively.

[0314] The imaging results are shown in the accompanying Figure 17 ( 68 Ga-Formula II-2-4-BD-WJ-DOTA), the accompanying Figure 18 ( 68 Ga-Formula II-2-4-BD-WJ-DOTA), and the accompanying Figure 19 ( 68 Ga-Formula II-2-4-BD-WJ-DOTA).

[0315] The results show that: 68 Ga-Formula II-2-4-BD-WJ-DOTA,68 Ga-Formula II-2-4-BD-WJ-DOTA and 68 Ga-Formula II-2-4-PN-WJ-DOTA showed a rapid accumulation in PSMA-positive LNCaP tumors at 10 min, and all images showed a clean background at 120 min. Under the same parameters, 68 Ga-Formula II-1-5-BD-WJ-DOTA showed higher tumor uptake than other radioligands, while its heart uptake was most pronounced at 10 min, followed by rapid washout and near background values at 30 min. In addition, only the kidneys and bladder could significantly take up these radioligands except for the tumor, indicating that these drugs were excreted into the body through the kidneys.

[0316] Test Example 10

[0317] This test example discloses 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-PN-WJ-DOTA and 177 The LNCaP animal model SPECT / CT imaging experiment of Lu-Formula II-2-4-PN-WJ-DOTA, specifically:

[0318] LNCaP cells were implanted in the left anterior limb of NOD / SCID mice under the armpit, and the SPF level animal room was bred for about 4 weeks, and the tumor mass was about 0.8 cm in diameter. About 100 μL of high-activity pure drug with a dose of about 200 μCi (radiochemical purity > 98.5%) was injected through the tail vein. Imaging was performed at 1 hour, 4 hours, 24 hours, 48 hours and 96 hours, respectively. The imaging results are shown in the accompanying Figure 24-26 177 Lu-Formula II-1-5-BD-WJ-DOTA), the accompanying Figure 24 177 Lu-Formula II-2-4-BD-WJ-DOTA) and the accompanying Figure 25 177 Lu-Formula II-2-4-PN-WJ-DOTA).

[0319] The results show that: 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-PN-WJ-DOTA and 177 Lu-Formula II-2-4-PN-WJ-DOTA maintained a good tumor-to-background ratio within 96 hours. The tumor showed obvious radioactive signals at 1 hour, and the rest of the radioactive accumulation was in the kidneys and bladder. 177 Lu-Formula II-2-4-PN-WJ-DOTA and 177 ​​​The uptake of Lu-Formula II-2-4-PN-WJ-DOTA at the tumor reached a peak at 4 hours, and then gradually decreased over time. 177 The peak time of tumor uptake of Lu-Formula II-1-5-BD-WJ-DOTA was about 48 hours after injection. In terms of kidney absorption, 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-PN-WJ-DOTA and 177 Lu-Formula II-2-4-PN-WJ-DOTA all showed rapid clearance.

[0320] Test Example 11

[0321] This test example discloses the pharmacodynamics test of the compound of the present application.

[0322] The drugs used in this test example are as follows:

[0323] 177 Lu-Formula II-1-5-BD-WJ-DOTA was prepared according to the method of Example 9.

[0324] 177 Lu-Formula II-2-4-BD-WJ-DOTA was prepared according to the method of Example 9.

[0325] 177 Lu-Formula II-2-4-PN-WJ-DOTA was prepared according to the method of Example 9. Positive control drug:

[0326] 177 Lu-PSMA-617 (Aladdin, 25mg), which became the first FDA-approved radioligand for internal radiotherapy of prostate cancer in 2022, prolongs the progression-free survival and overall survival of patients with PSMA-positive metastatic castration-resistant prostate cancer.

[0327] 177 Lu-Flu-1: This compound is the compound in the applicant's prior application, Patent "Prostate-specific membrane antigen inhibitor, its radionuclide marker and preparation method and application" with publication number CN115010629A 177 Lu-Formula II-1-SP-W0-DOTA, the compounds involved in this patent are all modified on the basis of the molecular structure of this compound, so it is used as a control drug for efficacy research.

[0328] Negative control: normal saline

[0329] The specific experimental steps are as follows:

[0330] LNCaP cells were implanted in the left anterior limb axillary of NOD / SCID mice, which were bred in SPF level animal room. When the average tumor volume of LNCaP tumor-bearing mice reached about 250 ± 25 mm 3 , the in vivo distribution and radioligand therapy studies were carried out.

[0331] 1. In vivo distribution: Male mice carrying LNCaP tumors with an average weight of about 20 ± 5 g were injected with 2.5 MBq of the corresponding radioligand through the tail vein. Mice were sacrificed at 1, 4, 24, 48 and 96 h after injection, respectively, blood was drawn, organs of interest were quickly collected, dried and weighed. Radioactivity was detected using a gamma counter, and the biodistribution results were calculated as a percentage of injected dose per gram (%ID / g). Each time point consisted of at least 4 mice.

[0332] 2. Treatment: Tumor mice were randomly divided into 16 groups (7 in each group), as follows:

[0333] Saline group;

[0334] 177 Three dose groups of Lu-PSMA-617: 37 MBq, 18.5 MBq and 7.4 MBq;

[0335] 177 Three dose groups of Lu-Flu-1: 37 MBq, 18.5 MBq and 7.4 MBq;

[0336] 177 Three dose groups of Lu-BWD: 37 MBq, 18.5 MBq and 7.4 MBq;

[0337] 177 Three dose groups of Lu-P4-BWD: 37 MBq, 18.5 MBq and 7.4 MBq;

[0338] 177 Three dose groups of Lu-P4-PND: 37 MBq, 18.5 MBq and 7.4 MBq.

[0339] The formula for calculating the size of individual tumors (mm 3 ) is: (length x width 2 ) / 2. The tumor volume and body weight of all mice were monitored every two days. Once the body weight was reduced to 40% or the tumor volume exceeded 1500 mm 3 , the mice were euthanized.

[0340] Results:

[0341] 1. In vivo distribution

[0342] The in vivo distribution of drugs in each group is shown in the attached Figure 26, 20, 21, 22, 23.

[0343] From the above results it is known that: 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA, 177 Lu-Formula II-2-4-PN-WJ-DOTA, 177 Lu-PSMA-617, 177 Lu-Flu-1 showed a higher tumor radioactivity at 1 h after injection and reached a peak at 4 h, followed by a gradual decrease in tumor uptake values, but overall maintained a high tumor uptake over 96 h. In comparison to 177 Lu-PSMA-617: 177 Lu-Formula II-1-5-BD-WJ-DOTA and 177 Lu-Formula II-2-4-BD-WJ-DOTA showed a faster tumor targeting uptake at early time points, within 48 h, 177 Lu-Formula II-1-5-BD-WJ-DOTA showed significantly higher tumor uptake values than 177 Lu-PSMA-617 until 96 h, 177 Lu-Formula II-1-5-BD-WJ-DOTA showed only slightly lower tumor uptake than 177 Lu-PSMA-617; 177 Lu-Formula II-2-4-BD-WJ-DOTA showed higher tumor uptake values than 177 Lu-PSMA-617 within 24 h, followed by slightly lower values than 177 Lu-PSMA-617 at later time points; 177 Lu-Formula II-2-4-PN-WJ-DOTA showed lower tumor uptake values than 177 Lu-PSMA-617 at all time points within 96 h. In comparison to 177 Lu-Flu-1, 177 Lu-Formula II-1-5-BD-WJ-DOTA and 177 Lu-Formula II-2-4-BD-WJ-DOTA showed higher tumor uptake and longer retention of radioactivity at all time points; 177 Lu-Formula II-2-4-PN-WJ-DOTA showed comparable tumor uptake to 177 Lu-Flu-1 at 1 h, followed by lower values than 177 Lu-Flu-1 at later time points.

[0344] For kidney uptake, the results showed that the renal route is the main excretion route for all radioligands. High kidney radioactivity was observed at 1 h after intravenous injection, followed by a gradual decrease.177 Lu-Flu-1, 177 Lu-Formula II-1-5-BD-WJ-DOTA, 177 Lu-Formula II-2-4-BD-WJ-DOTA initially showed higher kidney uptake, but these values were far below 177 Lu-PSMA-617. 177 Lu-Formula II-1-5-BD-WJ-DOTA and 177 Lu-Formula II-2-4-BD-WJ-DOTA showed higher kidney uptake at 1 hour than 177 Lu-Flu-1, followed by a decrease to comparable levels, 177 Lu-Formula II-2-4-PN-WJ-DOTA showed lower kidney uptake from the beginning, with values far below 177 Lu-Flu-1. After 96 hours of injection, the accumulation of radioactivity in the kidneys decreased to lower levels for all drugs.

[0345] In addition, the accumulation of all radioligands in critical organs such as the heart was low after 4 hours of injection. The accumulation activity in the salivary glands, one of the most critical organs in PSMA-targeted radioligand therapy, was negligible for all radioligands investigated. Similarly, the accumulation activity in normal organs and tissues such as the liver, muscle, and blood, which had comparable low values in %ID / g, was negligible.

[0346] These results show that, 177 Lu-Formula II-2-4-PN-WJ-DOTA showed the lowest tumor uptake in combination with low kidney uptake. 177 Lu-Formula II-1-5-BD-WJ-DOTA and 177 Lu-Formula II-2-4-BD-WJ-DOTA had higher tumor uptake than 177 Lu-Flu-1 with fast kidney clearance, demonstrating a superior therapeutic potential. In particular, 177 Lu-Formula II-1-5-BD-WJ-DOTA demonstrated faster and higher tumor uptake and lower kidney uptake than 177 Lu-PSMA-617, and, therefore, 177 Lu-Formula II-1-5-BD-WJ-DOTA has similar or even better therapeutic potential than 177 Lu-PSMA-617.

[0347] 2. Therapeutic data

[0348] The therapeutic data for each dose group are shown in the attached ​ table.

[0349] The attached ​At a dose of 37 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H).

[0350] Among them, the appendix ​ At a dose of 18.5 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H).

[0351] Among them, the appendix ​ At a dose of 7.4 MBq, the overall trend of tumor volume change (A) and the trend of body weight change (B) in each group of animals; the trend of individual tumor changes in each treatment group at the corresponding dose (C-H).

[0352] The results above show that all treated mice experienced a slight decrease in body weight, but this weight loss decreased or even returned to normal levels when the tumor volume was controlled. 177 37 MBq of Lu-type II-1-5-BD-WJ-DOTA. Overall, throughout the treatment period, all treated mice maintained a weight of more than 65% of their initial body weight.

[0353] Tumors grew rapidly in all mice in the saline group, exceeding 1500 mm in volume within 30 days. 3 Compared with the control group, single doses of 37 MBq, 18.5 MBq, or 7.4 MBq... 177 Lu-PSMA-617, 177 Lu-Flu-1, 177 Lu-style II-2-4-BD-WJ-DOTA 177 Lu-style II-2-4-PN-WJ-DOTA and 177 The Lu-type II-1-5-BD-WJ-DOTA treatment group all showed significant inhibition or delay in tumor growth.

[0354] 37 MBq and 18.5 MBq dose groups: During a 60-day treatment cycle, 177 Lu-PSMA-617, 177 Lu-Flu-1, 177 Lu-style II-2-4-BD-WJ-DOTA 177 Lu-style II-2-4-PN-WJ-DOTA and 177 In all Lu-type II-1-5-BD-WJ-DOTA treatment groups, tumor growth was significantly inhibited, and the tumor volume decreased to approximately 50 mm. 3 Even smaller. There were no significant differences in efficacy between the two dosage groups. In the 37MBq dosage group,177 Lu-formula II-1-5-BD-WJ-DOTA had the best recovery of body weight while tumors were inhibited or reduced in size in mice, better than 177 Lu-PSMA-617 and 177 Lu-Flu-1; 177 Lu-formula II-2-4-BD-WJ-DOTA, 177 Lu-formula II-2-4-PN-WJ-DOTA was better than 177 Lu-PSMA-617 and 177 Lu-Flu-1. In the 18.5 MBq dose group, 177 Lu-formula II-2-4-BD-WJ-DOTA, 177 Lu-formula II-2-4-PN-WJ-DOTA and 177 Lu-formula II-1-5-BD-WJ-DOTA had the best recovery of body weight while tumors were inhibited or reduced in size in mice, better than 177 Lu-PSMA-617 and 177 Lu-Flu-1.

[0355] 7.4 MBq dose group: 177 Lu-PSMA-617, 177 Lu-Flu-1, 177 Lu-formula II-2-4-BD-WJ-DOTA, 177 Lu-formula II-2-4-PN-WJ-DOTA had a significantly reduced inhibition of tumors, showing a short-term tumor inhibition in the tumor, followed by a trend of increasing tumor volume. Among them, 177 Lu-formula II-2-4-BD-WJ-DOTA, 177 Lu-formula II-2-4-PN-WJ-DOTA group reduced tumor volume within 20 days, followed by an increase in tumor volume. 177 Lu-PSMA-617, 177 Lu-Flu-1 group of tumors were inhibited within 30 days, followed by an increase in volume. 177 Lu-formula II-1-5-BD-WJ-DOTA still maintained a significant tumor inhibition, and the tumor volume decreased to about 50 mm 3 or even smaller. At this dose, 177 Lu-formula II-1-5-BD-WJ-DOTA had the best therapeutic effect in PSMA-positive LNCaP tumor-bearing mice.

[0356] Based on the above data, 177 Lu-formula II-1-5-BD-WJ-DOTA had the best therapeutic effect while ensuring body weight, indicating 177The therapeutic effect of Lu-type II-1-5-BD-WJ-DOTA in PSMA-positive LNCaP tumor-bearing mice is better than that of Lu-PSMA-617 177 Lu-PSMA-617 and 177 Lu-Flu-1.

[0357] In summary, the present application first prepared a novel macrocyclic polyamine carboxylic acid short peptide compound of formula I by two routes. The compound has mild preparation conditions, simple chemical reaction type, stable physical and chemical properties of the prepared compound. The labeling method of radionuclide label formula II is simple, efficient, and the reaction conditions are controllable. The labeling rate and radiochemical purity of the product are high, which can reach more than 99%. In the stability test, the labeled product has good stability in PBS and FBS. Formula I can be combined with diagnostic nuclides 68 Ga, 64 Cu, 18 F, etc. to form imaging agents for targeted diagnosis, and can be labeled with beta particle emitter nuclides 177 Lu, 225 Ac, etc. for tumor nuclide targeted 68 Ga, 64 Cu, internal irradiation therapy.

[0358] When formula I is labeled with nuclides such as 68 Ga and 64 Cu for targeted diagnosis, in vivo PET / CT imaging studies, it is found that 68 Ga-type II and 64 Cu-type II have good tumor uptake in LNCaP tumor models, and the tumor-to-kidney, muscle, and other organ ratios are high. In LNCaP tumor models, they have good imaging performance and have the potential to become a new generation of prostate cancer imaging and guidance imaging agents.

[0359] When formula I is labeled with nuclides such as 177 Lu and 225 Ac for tumor nuclide targeted internal irradiation therapy, the obtained 177 Lu-type II and 225 Ac-type II show excellent 24-hour PBS and serum stability in physicochemical property and in vivo and in vitro property studies. SPECT imaging and biodistribution studies show that the label is mainly excreted through the kidney, and the drug shows high affinity for PSMA in LNCaP models, which to some extent improves tumor absorption and treatment effect, and is worthy of further research on prostate cancer internal radiotherapy.

[0360] The embodiments described above are only part of the embodiments of the present application, rather than all the embodiments. The detailed description of the embodiments of the present application is not intended to limit the scope of the claimed application, but only to represent selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor are within the scope of protection of the present application.

Claims

1. A prostate specific membrane antigen inhibitor as shown in formula I, R1＝ ； R2＝ ； A＝ ； B＝ ； C＝ ； D= 0.5 wherein X is alkyl or halogen; E＝ ； wherein: when a = b = e = 0, d = 1, c = an integer from 1 to 8; when c = e = 0, a = d = 1, b = an integer from 1 to 8; when c = d = 0, a = e = 1, b = an integer from 1 to 8.

2. The PSMA inhibitor according to claim 1, characterized in that X is C1-C4 alkyl or halogen.

3. The PSMA inhibitor according to claim 2, characterized in that The halogen is I.

4. The PSMA inhibitor according to claim 3, characterized in that The alkyl is methyl.

5. The radiolabeled agent of any one of claims 1 to 4, wherein the agent is a prostate-specific membrane antigen inhibitor. The structure is shown in formula II: R3＝ ； wherein M is a radionuclide; R1, A, B, C, D, E, X, a, b, c, d, e are defined as in claim 1.

6. The radiolabeled inhibitor of prostate-specific membrane antigen according to claim 5, wherein, M= 68 Hey, 177 Monday, 225 Needle 64 With.

7. Process for the preparation of a compound of formula I according to any one of claims 1 to 4, characterized in that comprising the following steps: Step 1. Compound b6 and b7-Dde undergo nucleophilic substitution reaction to generate compound b8; Step 2. Compound b8 removes the protecting group to generate compound b9; Step 3. Compound b9 and b10 undergo polypeptide coupling reaction to generate compound b11; Step 4. Compound b11 removes the protecting group to generate compound formula I; The reaction scheme is as follows: ； wherein R1, R2, A, B, C, D, E, X, a, b, c, d, e are defined as in claim 1.

8. Process for the preparation of a compound of formula I according to claim 7, characterized in that In step 1, compound b6 and compound b7-Dde are coupled to generate compound b8 in the presence of a basic solvent and a condensing agent; Or / and in step 2, compound b8 is reduced with a reducing agent in a solvent to generate compound b9; Or / and in step 3, compound b9 and compound b10 are coupled to generate compound b11 in the presence of a basic solvent and a condensing agent; Or / and in step 4, compound b11 is deprotected in an acidic solvent to generate compound formula I; The solvent in steps 1-4 is an aprotic polar solvent; The reaction temperature in steps 1-4 is 0-60℃, and the reaction time is 1-24 hours.

9. Process for the preparation of a compound of formula I according to claim 8, characterized in that In step 1, the molar ratio of compound b7-Dde to compound b6 is 1.0-2.

0.

10. Process for the preparation of a compound of formula I according to claim 8, characterized in that In step 3, the molar ratio of compound b9 to compound b10 is 1.0-2.

0.

11. Process for the preparation of a compound of formula I according to claim 8, characterized in that In steps 1 and 3, the base is at least one of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, and cyclohexylamine.

12. Process for the preparation of a compound of formula I according to claim 8, characterized in that In steps 1 and 3, the molar ratio of the base to b6 or b9 is 1.0-5.

0.

13. Process for the preparation of a compound of formula I according to claim 8, characterized in that In steps 1 and 3, the condensing agent used in the coupling reaction is at least one of HATU, HBTU, HOBT, DCC, and EDCI.

14. Process for the preparation of a compound of formula I according to claim 8, characterized in that The solvent in steps 1-4 is at least one of dichloromethane, chloroform, tetrahydrofuran, 1,4-dioxane, and acetonitrile.

15. Process for the preparation of a compound of formula I according to claim 7, characterized in that The preparation method of compound b6 comprises the following steps: S1-1. Mannich reaction of compound b1 and b2-1 to generate compound b3-1; S1-2. Nucleophilic substitution reaction of compound b3-1 and b4 to generate compound b5-1; S1-3. Azide reduction of compound b5-1 to generate compound b6; The reaction scheme is as follows: wherein R1, A, B, C, a, b, c, are defined as in claim 1, and n is an integer from 1 to 6.

16. Process for the preparation of a compound of formula I according to claim 15, characterized in that In the S1-1, the Mannich reaction of the compound b1 with the compound b2-1 and CH3BNNa in an organic solvent generates the compound b3-1.

17. Process for the preparation of a compound of formula I according to claim 16, characterized in that The molar ratio of the compound b1 to the compound b2-1 is 1.0-3.

0.

18. Process for the preparation of a compound of formula I according to claim 16, characterized in that In the S1-1, the molar ratio of the compound b1 to CH3BNNa is 1.0-3.

0.

19. Process for the preparation of a compound of formula I according to claim 16, characterized in that The organic solvent used in the S1-1 is a polar solvent.

20. The method for preparing the compound of formula I according to claim 16, characterized in that, The organic solvent used in the S1-1 is any one or several of dimethylformamide, methanol, ethanol, water, formic acid, acetic acid, and hydrochloric acid.

21. A process for the preparation of a compound of formula I according to claim 16, characterized in that The reaction temperature in the S1-1 is 0-80°C, and the reaction time is 4-24 hours.

22. The process of claim 15, wherein the process is for the preparation of a compound of formula I, ###00011### I In the S1-2, the nucleophilic substitution reaction of the compound b3-1 with b4 in a basic organic solvent generates the compound b5-1.

23. Process for the preparation of a compound of formula I according to claim 22, characterized in that The molar ratio of the compound b3-1 to the compound b4 is 1.0-3.

5.

24. Process for the preparation of a compound of formula I according to claim 22, characterized in that In the S1-2, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, and cyclohexylamine, and the molar ratio of the base to the compound b3-1 is 1.0-5.

0.

25. The process according to claim 22 for the preparation of a compound of formula I, wherein The organic solvent in the S1-2 is an aprotic polar solvent.

26. The method for preparing the compound of formula I according to claim 22, characterized in that, The organic solvent in the S1-2 is any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, and acetonitrile.

27. The method for preparing the compound of formula I according to claim 22, characterized in that, The reaction temperature in the S1-2 is 0-60°C, and the reaction time is 4-48 hours.

28. A process for the preparation of a compound of formula I according to claim 15, wherein In the S1-3, the reduction reaction of the compound b5-1 with a reducing agent in a basic organic solvent generates b6.

29. Process for the preparation of a compound of formula I according to claim 28, characterized in that In the S1-3, the reducing agent is Pd / C and hydrogen or triphenylphosphine, the amount of Pd / C is 1.25-20.50% of the molar amount of the compound b5-1, the amount of hydrogen is 1.0-20.0% of the molar amount of the compound b5-1, and the amount of triphenylphosphine is 1.0-5.0 times the molar amount of the compound b5-1.

30. The method for preparing the compound of formula I according to claim 28, characterized in that, The organic solvent in the S1-3 is a polar solvent.

31. A process for the preparation of a compound of formula I according to claim 28, wherein The organic solvent in the S1-3 is any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile, water, methanol, and ethanol.

32. The method for preparing the compound of formula I according to claim 28, characterized in that, The reaction temperature in the S1-3 is 0-90°C, and the reaction time is 1-20 hours.

33. A process for the preparation of a compound of formula I according to claim 7, characterized in that The preparation method of the compound b6 comprises the following steps: S2-1. Mannich reaction of the compound b1 with b2-2 generates the compound b3-2; S2-2. Nucleophilic substitution reaction of the compound b3-2 with b4 generates the compound b5-2-1; S2-3. Compound b5-2-1 is deprotected to generate the compound b5-2-2; S2-4. Polypeptide coupling reaction of the compound b5-2-2 with b5-2-3 generates the compound b5-2-4; S2-5. Compound b5-2-4 is deprotected to generate the compound b6; The reaction route is shown in the following scheme: wherein R1, A, B, C, a, b, c, and n are as defined in claim 1, and n is an integer of 1-6.

34. Process for the preparation of a compound of formula I according to claim 33, characterized in that In the S2-1, the Mannich reaction of the compound b1 with the compound b2-2 and NaBH4 in an organic solvent generates the compound b3-2.

35. Process for the preparation of a compound of formula I according to claim 34, characterized in that The molar ratio of the compound b1 to the compound b2-2 is 1.0-3.

0.

36. The method for preparing the compound of formula I according to claim 34, characterized in that, The molar ratio of compound b1 to NaBH4 is 1.0-3.

0.

37. The method for preparing the compound of formula I according to claim 34, characterized in that, The organic solvent in S2-1 is a polar organic solvent.

38. The method for preparing the compound of formula I according to claim 34, characterized in that, The organic solvent in S2-1 is any one or several of dimethylformamide, methanol, ethanol, water, formic acid, acetic acid, and hydrochloric acid.

39. The method for preparing the compound of formula I according to claim 34, characterized in that, The reaction temperature of S2-1 is 0-80°C, and the reaction time is 4-24 hours.

40. The method for preparing the compound of formula I according to claim 33, characterized in that, In S2-2, compound b3-2 reacts with b4 in a basic organic solvent to generate compound b5-2-1 through nucleophilic substitution.

41. A process for the preparation of a compound of formula I according to claim 40, wherein The molar ratio of compound b3-2 to compound b4 is 1.0-3.

5.

42. The method for preparing the compound of formula I according to claim 40, characterized in that, In S2-2, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, and cyclohexylamine, and the molar ratio of the base to compound b3-2 is 1.0-5.

0.

43. The method for preparing the compound of formula I according to claim 40, characterized in that, The organic solvent in S2-2 is an aprotic polar solvent.

44. The method for preparing the compound of formula I according to claim 40, characterized in that, The organic solvent in S2-2 is any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, and acetonitrile.

45. The method for preparing the compound of formula I according to claim 40, characterized in that, The reaction temperature in S2-2 is 0-60°C, and the reaction time is 4-48 hours.

46. ​​The method for preparing the compound of formula I according to claim 33, characterized in that, In S2-3, compound b5-2-1 is deprotected under basic solvent conditions to generate compound b5-2-2.

47. The method for preparing the compound of formula I according to claim 46, characterized in that... In S2-3, the base is one or more of lithium hydroxide, sodium hydroxide, potassium hydroxide, and barium hydroxide, and the molar ratio of the base to compound b5-2-1 is 1.0-5.

0.

48. The method for preparing the compound of formula I according to claim 46, characterized in that, In S2-3, the solvent is a protic polar solvent.

49. The method for preparing the compound of formula I according to claim 46, characterized in that, In S2-3, the solvent is any one or several of water, methanol, ethanol, isopropanol, and n-butanol.

50. The method for preparing the compound of formula I according to claim 46, characterized in that, The reaction temperature in S2-3 is 0-90°C, and the reaction time is 1-20 hours.

51. The method for preparing the compound of formula I according to claim 33, characterized in that, In S2-4, compound b5-2-2 reacts with compound b5-2-3 in the presence of a basic solvent and a condensing agent to generate compound b5-2-4 through coupling reaction.

52. The method for preparing the compound of formula I according to claim 51, characterized in that, The molar ratio of compound b5-2-3 to compound b5-2-2 is 1.0-3.

0.

53. The method for preparing the compound of formula I according to claim 51, characterized in that, In S2-4, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, and cyclohexylamine, and the molar ratio of the base to compound b5-2-2 is 1.0-5.

0.

54. The method for preparing the compound of formula I according to claim 51, characterized in that, In S2-4, the coupling agent used is one or more of HATU, HBTU, HOBT, DCC, and EDCI, and the molar ratio of the coupling agent to compound b5-2-2 is 1.0-5.

0.

55. The method for preparing the compound of formula I according to claim 51, characterized in that, In S2-4, the solvent is an aprotic polar solvent.

56. The method for preparing the compound of formula I according to claim 51, characterized in that, In S2-4, the solvent is any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, and acetonitrile.

57. The method of claim 51, wherein the compound of formula I is prepared by the process of: ###00020### I 57 The reaction temperature of S2-4 is 0-60°C, and the reaction time is 1-24 hours.

58. The method for preparing the compound of formula I according to claim 33, characterized in that, In S2-5, compound b5-2-4 is deprotected under basic solvent conditions to generate compound b6.

59. The method for preparing the compound of formula I according to claim 58, characterized in that, In S2-5, the base is one or more of triethylamine, diethylamine, diethanolamine, pyridine, diisopropylamine, ethylenediamine, and cyclohexylamine, and the molar ratio of the base to compound b5-2-4 is 1.0-5.

0.

60. The method of claim 58, wherein the compound of formula I is prepared by the process of: ###0007### I In the S2-5, the solvent is a non-protic polar solvent.

61. The method for preparing the compound of formula I according to claim 58, characterized in that, In the S2-5, the solvent is any one or several of dichloromethane, trichloromethane, tetrahydrofuran, 1,4-dioxane, acetonitrile.

62. The method for preparing the compound of formula I according to claim 58, characterized in that, The reaction temperature of S2-5 is 0-90℃, and the reaction time is 1-24 hours.

63. A method of preparing a radiolabelled PSA inhibitor according to claim 5 or 6, wherein, The compound of formula I is reacted with a radioactive metal salt to generate a radioactive label of the compound of formula I, formula II, and the reaction formula is as follows: wherein R1, R 2、 R 3、 A, B, C, D, E, a, b, c, d, e are each as defined in claim 5.

64. Use of the compound of formula I according to any one of claims 1-4 or the radioactive label of the prostate-specific membrane antigen inhibitor according to claim 5 or 6 in the preparation of a prostate cancer diagnostic reagent / drug or / and a therapeutic drug.

Images