Radiolabeled FAPI complex, preparation method therefor, and use thereof

Abstract

The present invention belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine diagnosis and treatment and relates to a radiolabeled FAPI complex, a preparation method therefor, and a use thereof. The general structural formula of FAPI capable of realizing radionuclide labeling is as shown below: wherein R is a nitroimidazole group or -COOH; X is a chelating group or a chelating structure chelated with a radionuclide; L1 and L2 are linking groups; and A is an adjusting atom of an FAP-targeting group, and all the A atoms are H atoms or F atoms. The synthesis method for a radionuclide-labeled precursor of the present invention is simple and suitable for various nuclide labels, the labeling method is convenient, and the label has high radiochemical purity and good stability. The radiolabeled FAPI complex shows high affinity and specificity for FAP in a cell experiment and a high tumor uptake and target to non-target ratio in a tumor-bearing mouse, and is a potential FAP-targeting radioactive diagnosis and treatment drug.

Description

A radiolabeled FAPI complex and its preparation method and application Technical Field

[0001] The present invention relates to a novel radionuclide-labeled complex and a preparation method and application thereof, and in particular to a radioactively labeled FAPI complex and a preparation method and application thereof, belonging to the fields of radiopharmaceutical chemistry technology and nuclear medicine diagnosis and treatment technology. Background Art

[0002] Fibroblast activation protein (FAP) is absent or expressed at low levels in healthy adult tissues. However, its expression increases in non-malignant diseases involving tissue remodeling, such as wound healing, cirrhosis, arthritis, atherosclerosis, and fibrosis, and correlates with disease progression. Importantly, FAP is highly expressed in stromal cancer-associated fibroblasts in over 90% of epithelial cancers. Therefore, FAP is a potential target for tumor detection and treatment.

[0003] Radionuclide-labeled quinoline-based small molecule inhibitors targeting fibroblast activation protein (FAPI) have shown promising results in the diagnosis of tumors and other diseases. Among them, FAPI-04 is the most representative, which has good affinity and specificity for FAP. 68 Ga]Ga-FAPI-04 is rapidly cleared from the body through the kidneys and exhibits a good tumor-to-background ratio in PET imaging. FAPI-04 can also be used to label therapeutic radionuclides such as Lu-177. However, for radioactive therapeutic drugs, FAPI-04 tumor uptake values ​​are relatively low, and in particular, tumor retention time still needs to be improved. Therefore, further increasing drug uptake and retention time in tumors while reducing uptake in non-target tissues is the current direction and goal of FAPI drug development.

[0004] Since 2018, a large number of studies have optimized the non-pharmacophore portion of the FAPI structure, but these results have limited improvements in the tumor uptake dose and retention time of FAPI, and cannot fully meet the needs of clinical diagnosis or treatment. In 2021, the inventors developed a Ga-68-labeled FAPI-type radioactive molecular probe (patent number: ZL202111437982.7). This type of drug has high affinity and specificity for FAP and shows high tumor uptake in tumor-bearing mice. However, this type of drug is metabolized through the liver and intestinal system, with high uptake and slow clearance in the liver, gallbladder and intestines, and there is still room for optimization.

[0005] The FAPIs reported so far all have low target uptake and are rapidly eluted, resulting in a low target-to-non-target ratio, which is not conducive to disease diagnosis, imaging, and treatment.

[0006] Therefore, a fibroblast activation protein inhibitor, FAPI, and its preparation and application are provided. The synthesis method is simple, amenable to multi-nuclide labeling, and the labeling method is convenient. The labeled product has high radiochemical purity and good stability. The radionuclide-labeled FAPI complex exhibited high affinity and specificity for FAP in cell-based experiments and demonstrated high tumor uptake and target-to-nontarget ratios in tumor-bearing mice. This novel FAPI, characterized by rapid and efficient uptake and long-term retention in target tissues and rapid clearance from non-target organs, could increase the drug's uptake dose and retention time at the target site, while reducing damage to the blood and non-target organs. It is a potential FAP-targeted radioactive diagnostic and therapeutic drug.

[0007] Summary of the Invention

[0008] One of the objectives of the present invention is to develop a novel radiolabeled FAPI complex that effectively combines with the FAPI group through different chelating groups and linking groups, thereby increasing the drug uptake in tumors and extending the retention time, thereby overcoming the shortcomings of radiolabeled FAPI compounds currently reported in the literature, such as low tumor uptake values ​​and short retention times, which are not conducive to clinical diagnosis and treatment of tumors. The novel radiolabeled FAPI complex has a simple synthesis method, is suitable for labeling with a variety of radionuclides, and has a convenient labeling method. The labeled compound has high radiochemical purity and good stability. The radiolabeled FAPI complex exhibits high affinity and specificity for FAP in cell experiments, high tumor uptake values ​​and long tumor retention times in tumor-bearing mice, and an excellent target-to-non-target ratio. It is a potential new FAP-targeted radioactive diagnostic and therapeutic drug.

[0009] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0010] A radiolabeled FAPI complex has the following general structural formula:

[0011] Wherein, R is a nitroimidazole group or -COOH, and the nitroimidazole group is selected from any one of the following:

[0012] X is a chelating group or a chelating structure for chelating radionuclides, selected from any one of the following:

[0013] Among them, M includes but is not limited to 67 Ga 3+ 、 68 Ga 3+ 、[Al 18 F] 2+ 、 64 Cu 2+ 、 67 Cu2+ 、 111 In 3+ 、 177 Lu 3+ 、 86 Y 3+ 、 90 Y 3+ 、 44 Sc 3+ 、 47 Sc 3+ 、 225 Ac 3+ 、 212 Pb 2+ 、 203 Pb 2+ 、 213 Bi 3+ 、 212 Bi 3+ wait;

[0014] L1 and L2 are linking groups, wherein L1 is selected from any one of the following:

[0015] In the formula, n is an integer from 0 to 6;

[0016] L2 is selected from any of the following:

[0017] A is the regulatory atom of the FAP targeting group, and A is either an H atom or an F atom.

[0018] Preferably, the specific structure is:

[0019] Another object of the present invention is to provide a method for preparing the radiolabeled FAPI complex.

[0020] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0021] A preparation of a radiolabeled FAPI complex comprises the following steps:

[0022] (1) Nitroimidazole and anhydrous potassium carbonate were dissolved in ultra-dry N,N-dimethylformamide and stirred at room temperature. N-(3-bromopropyl)carbamic acid tert-butyl ester was added. After the reaction was completed, the mixture was filtered and extracted. The organic phase was dried and purified to obtain a yellow-green oily compound. The compound was deprotected with trifluoroacetic acid to obtain intermediate 1 as a white solid.

[0023] (2) (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid was dissolved in ultra-dry N,N-dimethylformamide. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice-cooling. The mixture was stirred and the white solid intermediate 1 obtained in step (1) was added. After the reaction was completed, the mixture was extracted, the organic phase was dried, and purified to obtain a light yellow solid. The yellow solid was deprotected with trifluoroacetic acid to obtain a yellow oil, which was dissolved in N,N-dimethylformamide. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, stirred, and FAPI (specifically (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-dimethylamino)propoxy]-1-piperazinyl ... Quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5 -diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester), after the reaction is completed, extraction, the organic phase is dried and purified to obtain a light yellow oil, which is deprotected with diethylamine to obtain a light yellow oily intermediate 2 (NI-FAPI);

[0024] (3) FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamyl) 1-Heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylate) were dissolved in trifluoroacetic acid, stirred at room temperature, and the solvent was evaporated to obtain a yellow oily substance. The yellow oily substance was dissolved in anhydrous acetonitrile, anhydrous potassium carbonate was added, and the mixture was stirred at room temperature. PEG chain (BocNH(CH2CH2O)) was added under ice-cooling. n CH2CH2Br), reflux reaction, overnight, filter, evaporate the solvent, purify, and obtain light yellow oily intermediate 3 (PEG n -FAPI);

[0025] (4) The chelating agent is dissolved in ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are added under ice bath, stirred, and N-Cbz-1,2-diaminoethane is added. After the reaction is completed, the mixture is extracted with ethyl acetate and saturated brine, and the organic phase is dried over anhydrous sodium sulfate and purified by silica gel column chromatography to obtain a yellow solid. The yellow solid is deprotected with hydrogen to obtain a white solid substance; 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,1 2-Dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene)bis(4-hydroxy-3,1-phenylene)dipropionic acid (HBED-CC(tBu)2) was dissolved in ultra-dry N,N-dimethylformamide. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice-cooling. The mixture was stirred and a white solid substance was added. After the reaction was completed, the mixture was extracted, the organic phase was dried, and purified to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu)2).

[0026] (5) The chelating agent is dissolved in ultra-dry N,N-dimethylformamide, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are added under ice bath, stirred, and the intermediate 2 prepared in step (2) or FAPI after deprotection of the Boc protecting group by trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-( tert-Butyl 3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate, (S)-tert-butyl 4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylate); after completion of the reaction, extraction was performed, the organic phase was dried, and purified to obtain a light yellow oil, which was deprotected with trifluoroacetic acid and purified by semi-preparative HPLC to obtain Chelator-NI-FAPI or Chelator-FAPI labeled precursor;

[0027] (6) The chelating agent is dissolved in ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are added under ice bath, stirred, and the intermediate 1 prepared in step (1) is added, and the reaction is carried out at room temperature overnight. After the reaction is completed, the organic phase is extracted and dried, and purified to obtain a yellow oily substance, which is dissolved in ultra-dry N,N-dimethylformamide and 2-( 7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were stirred, and FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4- Boc-1-piperazinyl)propyloxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4 -(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester), react at room temperature overnight. After the reaction, extract, dry the organic phase, and purify to obtain a light yellow oil. Deprotect with trifluoroacetic acid and purify by semi-preparative HPLC to obtain NI-Chelator-FAPI labeled precursor;

[0028] (7) The intermediate 4 prepared in step (4) was dissolved in N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, and stirred. FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-( (4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) or intermediate 3 prepared in step (3), after completion of the reaction, extraction, drying the organic phase, and purification to obtain a light yellow oily substance, which was deprotected with trifluoroacetic acid and purified by semi-preparative HPLC to obtain Chelator-HBED-CC-(PEG) n -FAPI labeled precursors;

[0029] (8) The labeled precursor obtained in steps (5), (6) and (7) is dissolved in dimethyl sulfoxide, sodium acetate buffer solution and a solution containing a radioactive nuclide are added, and the mixture is heated to obtain the corresponding radiolabeled product.

[0030] Preferably, in step (1), the nitroimidazole is 2-nitroimidazole, and the amount added is 1 equivalent; the amount added of anhydrous potassium carbonate is 3-5 equivalents; the amount added of ultra-dry N,N-dimethylformamide is 10-20 mL; and the amount added of tert-butyl N-(3-bromopropyl)carbamate is 1-2 equivalents.

[0031] Preferably, in step (1), diatomaceous earth is used for filtration, the filtrate is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified by silica gel column chromatography.

[0032] Preferably, in step (2), the amount of (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 10-20 mL; and the amounts of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine added are 1-2 equivalents.

[0033] Preferably, in step (2), the amount of the yellow oil added is 1-1.5 equivalents; the amount of the ultra-dry N,N-dimethylformamide added is 5-10 mL; the amount of the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine added is 1.3-1.5 equivalents; the FAPI after de-Boc protection by trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, The amount of the added components was 1 equivalent.

[0034] Preferably, in step (2), extraction is performed with ethyl acetate and saturated brine, and the organic phase is dried over anhydrous sodium sulfate and then purified by silica gel column chromatography.

[0035] Preferably, in step (3), the reaction is refluxed at 80° C.; potassium carbonate is removed by filtration using diatomaceous earth, the filtrate is evaporated under reduced pressure to remove the solvent, and the residue is purified by flash chromatography.

[0036] Preferably, in step (3), the FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide) The amount of (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) added is 1 equivalent; the amount of trifluoroacetic acid (TRIFLUOROACETIC ACID) added is 3-5 mL.

[0037] Preferably, in step (3), the yellow oily substance is added in an amount of 1 equivalent; the anhydrous potassium carbonate is added in an amount of 4-5 equivalents; and the PEG chain is tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamate, and the amount added is 1.2-1.5 equivalents.

[0038] Preferably, in step (4), the chelating agent includes 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-(tert-butoxy)-2-oxoethyl)-1,4-diazacycloheptane-6-yl)pentanoic acid (AAZTA(tBu)4), 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid (DOTA(tBu)3), 5-tert-butoxy-5-oxo-4-(4,7,10-tris(2-tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)pentanoic acid (DOTAGA(tBu)3), u)4), 4-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-7-(1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)-1,4,7,10-tetraazacyclododec-1-yl)-5-tert-butoxy-5-oxopentanoic acid (DOTA(GA)2(tBu)5), 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclopentan-1-yl)acetic acid (NOTA(tBu)2), 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazolidin-1-yl)-5-tert-butoxy-5-oxopentanoic acid (NODAGA(tBu)3).

[0039] Preferably, in step (4), the amount of the chelating agent added is 1 equivalent; the amount of the ultra-dry N,N-dimethylformamide added is 15-20 mL; the amount of the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1 equivalent; the amount of the N,N'-diisopropylethylamine added is 3 equivalents; and the amount of the N-Cbz-1,2-diaminoethane added is 1.2-1.5 equivalents.

[0040] Preferably, in step (4), extraction is performed with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified by silica gel column chromatography to obtain a yellow solid; the amount of HBED-CC(tBu)2 added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 10-20 mL; the amount of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1 equivalent; and the amount of N,N'-diisopropylethylamine added is 3 equivalents.

[0041] Preferably, in step (4), a white solid substance is added. After the reaction is completed, the mixture is extracted with ethyl acetate and saturated brine. The organic phase is dried over anhydrous sodium sulfate and purified by a speed purification chromatograph to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu)2)

[0042] Preferably, in step (5), the chelating agent is 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-(tert-butoxy)-2-oxoethyl)-1,4-diazacycloheptane-6-yl)pentanoic acid (AAZTA(tBu)4), 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane -1-yl)acetic acid (DOTA(tBu)3), 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu)2), 5-tert-butoxy-5-oxo-4-(4,7,10-tri( 2-tert-Butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanoic acid (DOTAGA(tBu)4), 4-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-7-(1,5-di-tert-butoxy-1,5-dioxolane-2-yl)-1,4,7,10-tetraazacyclododec-1-yl)-5-tert-butoxy-5-oxopentanoic acid ( DOTA(GA)2(tBu)5), 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclopentane-1-yl)acetic acid (NOTA(tBu)2), 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazolidin-1-yl)-5-tert-butoxy-5-oxopentanoic acid (NODAGA(tBu)3).

[0043] Preferably, in step (5), the amount of the chelating agent added is 1-1.2 equivalents; the amount of the ultra-dry N,N-dimethylformamide added is 2-5 mL; the amount of the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1-1.5 equivalents; the amount of the N,N'-diisopropylethylamine added is 1.5-3 equivalents; the intermediate 2 or the FAPI after de-Boc protection by trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2 -(2-cyanopyrrolidin-1-yl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, and (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) were added in an amount of 1 equivalent; after the reaction, the mixture was extracted with ethyl acetate and saturated brine, the organic phase was dried over anhydrous sodium sulfate, and purified using a speed purification chromatography instrument.

[0044] Preferably, in step (6), the chelating agent is 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu)2), 4,4'-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,7-diyl)bis(5-tert-butoxy-5-oxopentanoic acid) (DOTA(GA)2(tBu)4).

[0045] Preferably, in step (6), the amount of the chelating agent added is 1 equivalent; the amount of the ultra-dry N,N-dimethylformamide added is 2-10 mL; the amount of the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1 equivalent; the amount of the N,N'-diisopropylethylamine added is 3-5 equivalents; and the amount of the intermediate 1 added is 1 equivalent.

[0046] Preferably, in step (6), after the reaction is completed, the product is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a speed purification chromatograph.

[0047] Preferably, in step (6), the light yellow oil, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine and FAPI after de-Boc protection by trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide The amount of amine, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, and (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 7-10 mL.

[0048] Preferably, in step (6), after the reaction is completed, the product is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a speed purification chromatograph.

[0049] Preferably, in step (7), the amount of intermediate 4 added is 1 equivalent; the amount of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1-1.5 equivalents; the amount of N,N'-diisopropylethylamine added is 4-5 equivalents; the intermediate 3 or FAPI after de-Boc protection by trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1- The amount of addition of (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester and (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester is 1-1.5 equivalents.

[0050] Preferably, in step (7), after the reaction is completed, the product is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a speed purification chromatograph.

[0051] Preferably, in step (8), the solution containing radioactive nuclides is [ 68 Ga]GaCl3, [ 177 Lu]LuCl3, [ 18 F]AlF.

[0052] Another object of the present invention is to provide the application of the radiolabeled FAPI complex.

[0053] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0054] Application of radiolabeled FAPI complexes in FAP-targeted radiotherapy. Beneficial effects:

[0055] The novel radiolabeled FAPI complexes of the present invention effectively bind to FAPI groups through various chelating and linking groups, improving drug uptake and retention in tumors. The novel radiolabeled FAPI complexes are simple to synthesize and are suitable for labeling with a variety of radionuclides. The labeling method is convenient, and the labeled compounds exhibit high radiochemical purity and excellent stability. The radionuclide-labeled compounds have demonstrated high affinity and specificity for FAP in cell-based experiments, as well as high tumor uptake and target-to-nontarget ratios in tumor-bearing mice. They are promising FAP-targeted radioactive diagnostic and therapeutic drugs.

[0056] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but they are not intended to limit the scope of protection of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] FIG1 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-AAZTA-FAPI-04.

[0058] FIG2 is a graph showing the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-AAZTA-FAPI-04.

[0059] FIG3 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-AAZTA-NI-FAPI-04.

[0060] FIG4 is a graph showing the [ 177[Figure 5—Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-AAZTA-NI-FAPI-04.

[0061] Figure 5 is a diagram of the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-DOTA-NI-FAPI-04.

[0062] Figure 6 is a graph of the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of [Lu]Lu-DOTA-NI-FAPI-04.

[0063] FIG7 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-NI-FAPI-04.

[0064] FIG8 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-NI-HBED-CC-FAPI-04.

[0065] FIG9 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-NI-HBED-CC-FAPI-02.

[0066] FIG10 is a diagram of DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-FAPI-02.

[0067] FIG11 is a graph showing the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-02.

[0068] FIG12 is a DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-FAPI-04.

[0069] Figure 13 is a diagram of the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-04.

[0070] FIG14 is a DOTA-[ 68Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-PEG3-FAPI-02.

[0071] Figure 15 is a diagram of the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02.

[0072] FIG16 is a diagram of DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-PEG3-FAPI-04.

[0073] Figure 17 is a diagram of the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04.

[0074] FIG18 is a diagram of DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 Uptake-time curve of Ga]Ga-DOTA-FAPI-04 in HT1080-FAP cells in vitro.

[0075] FIG19 is a diagram of DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02(A), DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04(B)、DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02(C), DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04(D) and [ 68 Ga]Ga-DOTA-FAPI-04(E) uptake and blocking results in HT1080-FAP cells in vitro for 60 min.

[0076] FIG20 is a diagram of an embodiment 2 of the present invention. 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177 Uptake-time curve of Lu]Lu-DOTA-FAPI-04 in HT1080-FAP cells in vitro.

[0077] FIG21 is a diagram of an embodiment 2 of the present invention. 177 Lu]Lu-DOTA-HBED-CC-FAPI-02(A),[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04(B),[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02(C),[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04(D) and [ 177 [Lu]Lu-DOTA-FAPI-04(E) uptake and blocking results in HT1080-FAP cells in vitro for 60 min.

[0078] FIG. 22 is a diagram of an embodiment 3 of the present invention. 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 Uptake-time curve of Lu]Lu-DOTA-FAPI-04 in HT1080-FAP cells in vitro.

[0079] FIG23 is a diagram of an embodiment 3 of the present invention. 177 Lu]Lu-AAZTA-FAPI-04(F), [ 177 Lu]Lu-AAZTA-NI-FAPI-04(G), [ 177 Lu]Lu-DOTA-NI-FAPI-04(H) and [ 177 [Lu]Lu-DOTA-FAPI-04(E) uptake and blocking results in HT1080-FAP cells in vitro for 60 min.

[0080] FIG. 24 is a diagram of an embodiment 4 of the present invention. 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 Uptake-time curve of Ga]Ga-DOTA-FAPI-04 in HT1080-FAP cells in vitro.

[0081] FIG. 25 is a diagram of an embodiment 4 of the present invention. 68 Ga]Ga-HBED-CC-NI-FAPI-04(A),[ 68 Ga]Ga-NI-HBED-CC-FAPI-04(B),[ 68 Ga]Ga-NI-HBED-CC-FAPI-02(C) and [ 68 [Ga]Ga-DOTA-FAPI-04 (D) uptake and blocking results in HT1080-FAP cells in vitro for 60 min.

[0082] FIG26 is a diagram of an embodiment 5 of the present invention. 68 Ga]Ga-AAZTA-FAPI-04, [ 68 Ga]Ga-AAZTA-NI-FAPI-04, [ 68 Ga]Ga-DOTA-NI-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 In vivo PET / CT imaging results of U87MG tumor-bearing mice using Ga]Ga-DOTA-FAPI-04 (T: tumor; G: gallbladder). DETAILED DESCRIPTION

[0083] Unless otherwise specified, the reagents and raw materials used in the preparation and detection methods described in the following examples and comparative examples are all commercially available commodities, and the equipment used are all commonly used equipment; the concentrations and ratios described are all weight and amount units.

[0084] Example 1: 68 Preparation of Ga]Ga-AAZTA-FAPI-04

[0085] Step 1: Synthesis of AAZTA-FAPI-04

[0086] The synthetic route is as follows:

[0087] The specific steps include:

[0088] (1) Synthesis of Compound 1

[0089] In a 10 mL round-bottom flask, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2374782-82-6, 50 mg, 0.09 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was evaporated under reduced pressure and 3 mL of ultra-dry N,N-dimethylformamide was added. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (25.60 mg, 0.13 mmol), 1-hydroxybenzotriazole (18.00 mg, 0.13 mmol) and N,N-diisopropylethylamine (39.00 mg, 0.30 mmol) were added under ice bath. 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-tert-butoxy-2-oxoethyl)-1,4-diazacycloheptane-6-yl)pentanoic acid (AAZTA(tBu)4, 57.70 mg, 0.09 mmol) dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added, and the reaction solution was reacted at room temperature overnight. The reaction solution was diluted with ethyl acetate and washed with ethyl acetate (20 mL×3) and saturated brine (10 mL×1). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation of the filtrate under reduced pressure and purified by silica gel column chromatography (dichloromethane / methanol / ammonia water, v / v / v=20 / 1 / 0.1) to give orange-yellow compound 1 (64.70 mg, 0.06 mmol) in a yield of 67%.

[0090] Structure confirmation of compound 1:

[0091] HRMS C 58 H 88 N9O 12 F2[M+H] + Theoretical molecular weight is 1140.6526, and the measured molecular weight is 1140.6520.

[0092] 1H NMR (400MHz, CDCl3) δ8.82(d,J=4.4Hz,1H),8.04(d,J=9.2Hz,1H),7.50(d,J=4.3 Hz,1H),7.40(d,J=8.9Hz,1H),4.54-3.89(m,6H),3.61(s,4H),3.22(s,4H),2.99( d,J=13.9Hz,2H),2.80(dd,J=18.4,9.1Hz,4H),2.64(d,J=14.1Hz,5H),2.33(s,2H ),1.65-1.53(m,6H),1.44(d,J=3.8Hz,31H),1.25(s,14H),0.88(t,J=6.8Hz,4H).

[0093] (2) Synthesis of compound AAZTA-FAPI-04

[0094] In a 10 mL round-bottom flask, compound 1 (64.70 mg, 0.06 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was evaporated under reduced pressure and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4 × 250 mm) (Phase A: 0.1% trifluoroacetic acid-water, Phase B: 0.1% trifluoroacetic acid-acetonitrile, gradient: 0-12 min, 5%-25% B; 12-14 min, 25%-5% B; 14-15 min, 5% B, flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain AAZTA-FAPI-04 as a white solid. The purity was >97% as determined by LC-MS.

[0095] Structural confirmation of AAZTA-FAPI-04:

[0096] HRMS C 42 H 56 N9O 12 F2[M+H] + Theoretical molecular weight is 916.4011, and the measured molecular weight is 916.4016.

[0097] Step 2: Ga-68 labeling of AAZTA-FAPI-04:

[0098] The Ga-68 marked routes are as follows:

[0099] The marking method is as follows:

[0100] The AAZTA-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 9 μL of the precursor solution was taken and placed in a 10 mL vial. 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, and reacted at 50°C for 10 minutes. The radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector, and the radiochemical purity was greater than 95% [ 68 Ga]Ga-AAZTA-FAPI-04.

[0101] As shown in FIG1 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-AAZTA-FAPI-04 shows that [ 68 The radiochemical purity of Ga]Ga-AAZTA-FAPI-04 was greater than 95%.

[0102] Example 2: 177 Preparation of Lu]Lu-AAZTA-FAPI-04

[0103] Step 1: Synthesis of AAZTA-FAPI-04:

[0104] The synthetic route and method are the same as those in step 1 of Example 1;

[0105] Step 2: Lu-177 labeling of AAZTA-FAPI-04:

[0106] The Lu-177 marked route is as follows:

[0107] The marking method is as follows:

[0108] Prepare a 1 μg / μL precursor solution of AAZTA-FAPI-04 obtained in step 1 with dimethyl sulfoxide. Take 18 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 50 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-AAZTA-FAPI-04;

[0109] As shown in FIG2, the [ 177The radioactive HPLC spectrum of the labeled reaction solution of Lu]Lu-AAZTA-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-AAZTA-FAPI-04 was greater than 95%.

[0110] Example 3: 68 Preparation of [Ga]Ga-AAZTA-NI-FAPI-04)

[0111] Step 1: Synthesis of AAZTA-NI-FAPI-04:

[0112] The synthetic route is as follows:

[0113] The specific steps include:

[0114] (1) Synthesis of Compound 2

[0115] 2-Nitroimidazole (1.14 g, 10.09 mmol) was dissolved in 15 mL of ultra-dry N, N-dimethylformamide, anhydrous potassium carbonate (4.89 g, 35.38 mmol) was added, and the mixture was stirred at room temperature for 30 minutes to obtain a reaction solution. Tert-butyl N-(3-bromopropyl)carbamate (3.57 g, 14.99 mmol) dissolved in ultra-dry N, N-dimethylformamide (10 mL) was added to the reaction solution, and the mixture was stirred at room temperature overnight. The mixture was dried with diatomaceous earth. The mixture was filtered, and most of the solvent was removed by rotary evaporation under reduced pressure. The residue was washed with ethyl acetate (20 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and the anhydrous sodium sulfate solid was removed. The filtrate was rotary evaporation under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate, v / v = 1 / 1) to give a yellow-green oil 2 (2.36 g, 8.74 mmol) in a yield of 87%.

[0116] Structure confirmation of compound 2:

[0117] HRMS C 11 H 19 N4O4[M+H] + Theoretical molecular weight 271.1400, measured molecular weight 271.1408;

[0118] 1 H NMR (600MHz, CDCl3) δ7.27(s,1H),7.14(s,1H),4.75(s,1H),4.46(t,J=7.0Hz,2H),3.20(t,J=6.2Hz,2H),2.08-2.01(m,2H),1.44(s,9H);

[0119] (2) Synthesis of compound 3

[0120] Compound 2 (432 mg, 1.60 mmol) was dissolved in 4 mL of trifluoroacetic acid solution, stirred at room temperature for 30 minutes, and evaporated under reduced pressure to remove the solvent to obtain a white solid intermediate. (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid (1.5 g, 3.53 mmol) was dissolved in 10 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.61 g, 4.23 mmol) and N,N'-dimethylformamide were added under ice bath. Isopropylethylamine (547 mg, 4.23 mmol) was stirred in an ice bath for 25 minutes, and the above-mentioned white solid intermediate (603 mg, 3.55 mmol) was added. The mixture was stirred at room temperature overnight, washed with ethyl acetate (20 mL × 3) and saturated brine (20 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by silica gel column chromatography (dichloromethane / methanol / ammonia water, v / v / v = 25 / 1 / 0.1) to give compound 3 (1.5 g, 2.60 mmol) as a light yellow solid in a yield of 74%.

[0121] Structure confirmation of compound 3:

[0122] HRMS C 30 H 36 N5O7[M+H] + Theoretical molecular weight 578.2609, measured molecular weight 578.2609;

[0123] 1 H NMR (400MHz, CDCl3) δ7.76(d,J=7.6Hz,2H),7.59(t,J=7.2Hz,2H),7.40(t,J=7.5H z,2H),7.35-7.28(m,2H),7.27(s,1H),7.09(s,1H),6.57(s,1H),5.65(d,J=7.2Hz, 1H),4.41(dd,J=12.1,7.0Hz,4H),4.21(t,J=7.0Hz,2H),3.42-3.32(m,1H),3.31- 3.21(m,1H),2.35-2.18(m,3H),2.10-1.99(m,2H),1.92-1.81(m,1H),1.47(s,9H);

[0124] (2) Synthesis of compound 4

[0125] Compound 3 (319.6 mg, 0.55 mmol) was dissolved in 2 mL of dichloromethane, and then 2 mL of trifluoroacetic acid was added. The mixture was stirred at room temperature for 30 minutes. The reaction was detected by TLC and the solvent was removed by rotary evaporation under reduced pressure to obtain a yellow oily compound. The yellow oily compound (151.2 mg, 0.29 mmol) was dissolved in 2 mL of ultra-dry N,N-dimethylformamide. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (74.8 mg, 0.39 mmol), 1-hydroxybenzotriazole (52.7 mg, 0.39 mmol) and N,N'-diisopropylethylamine (134.4 mg, 1.04 mmol) were added under ice bath conditions. The mixture was stirred for 30 minutes under ice bath conditions to obtain a reaction solution. The product was deprotected by trifluoroacetic acid and dissolved in 2 mL of ultra-dry N,N (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2374782-82-6, 126.4 mg, 0.26 mmol) containing 1,2-dimethylformamide was added to the above reaction solution and reacted at room temperature overnight. The mixture was washed with ethyl acetate (20 mL × 3) and saturated brine (20 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and anhydrous sodium sulfate was removed. The filtrate was evaporated under reduced pressure to remove the solvent. The residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v = 15 / 1 / 0.1) to give compound 4 (205.6 mg, 0.21 mmol) as a light yellow oil in a yield of 81%.

[0126] Structure confirmation of compound 4:

[0127] HRMS C 50 H 54 N 11 O9F2[M+H] + Theoretical molecular weight 990.4068, measured molecular weight 990.4076;

[0128] 11H NMR (600 MHz, CD3OD) δ 8.69 (d, J = 4.4 Hz, 1H), 7.90 (dd, J = 16.1, 5.8 Hz, 2H), 7.73 (d, J = 7.5 Hz, 2H), 7.60 (dd, J = 11.1, 7.6 Hz, 2H), 7.51 (d, J = 4.4 Hz, 1H), 7.45 (s, 1H), 7.40 (dd, J = 9.2, 2.6 Hz, 1H), 7.32 (t, J = 7.4 Hz, 2H), 7.25 (ddd, J = 10.1, 5.5, 2.1 Hz, 2H), 7.03 (s, 1H), 5.07 (dd, J = 9.3, 2.7 Hz, 1H), 4.57 (dd, J = 9.7, 3.9 Hz, 1H), 4.44 - 4.37 (m, 2H), 4.34 (dd, J = 10.5, 6.9 Hz, 1H), 4.28 - 4.20 (m, 4H), 4.20 - 4.12 (m, 2H), 4.12 - 4.02 (m, 1H), 3.67 - 3.58 (m, 2H), 3.58 - 3.49 (m, 1H), 3.27 (dt, J = 3.1, 1.5 Hz, 3H), 3.15 - 3.07 (m, 1H), 2.85 (ddt, J = 14.0, 9.5, 6.9 Hz, 1H), 2.73 (t, J = 13.6 Hz, 1H), 2.61 - 2.44 (m, 5H), 2.32 - 2.20 (m, 2H), 1.98 (dtd, J = 20.6, 13.8, 6.9 Hz, 5H), 1.74 (td, J = 15.4, 6.0 Hz, 1H), 1.33 (dd, J = 26.1, 13.4 Hz, 1H), 1.29 - 1.21 (m, 3H);

[0129] (2) Synthesis of Compound 5

[0130] Compound 4 (50.5 mg, 0.05 mmol) was dissolved in 2 mL of dichloromethane, 1 mL of diethylamine was added, and the mixture was stirred at room temperature for 2 hours. The reaction was detected by TLC, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light yellow oily intermediate. 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-tert-butoxy-2-oxoethyl)-1,4-diazacycloheptane-6-yl)pentanoic acid (AAZTA(tBu)4, 42 mg, 0.06 mmol) was dissolved in 2 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (23 mg, 0.06 mmol) was added under ice bath. mol) and N,N'-diisopropylethylamine (13 mg, 0.1 mmol), stirred in an ice bath for 30 minutes to obtain a reaction solution, and a light yellow oily intermediate (38.37 mg, 0.05 mmol) dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution, reacted at room temperature overnight, and washed with ethyl acetate (10 mL×3) and saturated brine (10 mL×2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The filtrate was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain yellow oily compound 5 (29 mg, 0.02 mmol) in a yield of 40%;

[0131] HRMS C 69 H 103 N 14 O 16 F2[M+H] + Theoretical molecular weight 1421.7639, measured molecular weight 1421.7646;

[0132] 1H NMR(600MHz,CD3OD)δ8.75(d,J=4.1Hz,1H),7.96(dd,J=10.7,5.6Hz,2H),7.64-7.52(m,2H),7.46(d,J=9.1Hz,1H),7.1 3(s,1H),4.50(d,J=3.3Hz,2H),4.36-4.21(m,5H),4.15(dt,J=20.1,10.3Hz,1H),3.70(s,2H),3.66-3.51(m,5H),3.36- 3.24(m,7H),3.23-3.16(m,2H),2.94(dt,J=23.4,9.4Hz,2H),2.82(d,J=15.1Hz,2H),2.75-2.47(m,8H),2.37-2.23(m, 4H),2.15-2.00(m,5H),1.87-1.76(m,1H),1.63-1.56(m,2H),1.53(s,5H),1.46(d,J=11.0Hz,30H),1.38-1.25(m,12H);

[0133] (2) Synthesis of compound AAZTA-NI-FAPI-04

[0134] Compound 5 (29 mg, 0.02 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 2 hours. The reaction was detected by mass spectrometry and the solvent was removed by rotary evaporation under reduced pressure. The residue was dissolved in 2.9 mL of dimethyl sulfoxide and separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-25 min, 5%-100% B; 25-26 min, 100%-5% B; 26-30 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain AAZTA-NI-FAPI-04 as a white solid.

[0135] Structure confirmation of compound AAZTA-NI-FAPI-04:

[0136] HRMS C 53 H 70 N 14 O 16 F2[M+H] + Theoretical molecular weight 1197.5135, measured molecular weight 1197.5132;

[0137] Step 2: Ga-68 labeling of AAZTA-NI-FAPI-04:

[0138] The Ga-68 marked route is as follows:

[0139] The marking method is as follows:

[0140] The AAZTA-NI-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 6 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, and reacted at 50°C for 10 minutes. The radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector, and the radiochemical purity was greater than 95% [ 68 Ga]Ga-AAZTA-NI-FAPI-04;

[0141] As shown in FIG3, the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-AAZTA-NI-FAPI-04 shows that [ 68 The radiochemical purity of Ga]Ga-AAZTA-NI-FAPI-04 is greater than 95%;

[0142] Example 4: 177 Preparation of Lu]Lu-AAZTA-NI-FAPI-04

[0143] Step 1: Synthesis of AAZTA-NI-FAPI-04:

[0144] The synthetic route and method are the same as those in step 1 of Example 3;

[0145] Step 2: Lu-177 labeling of AAZTA-NI-FAPI-04:

[0146] The Lu-177 marked route is as follows:

[0147] The marking method is as follows:

[0148] Prepare a 1 μg / μL precursor solution of AAZTA-NI-FAPI-04 obtained in step 1 with dimethyl sulfoxide. Take 24 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 50 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-AAZTA-NI-FAPI-04;

[0149] As shown in FIG4, the [ 177 The radioactive HPLC spectrum of the labeled reaction solution of Lu]Lu-AAZTA-NI-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-AAZTA-NI-FAPI-04 was greater than 95%.

[0150] Example 5: 68 Preparation of [Ga]Ga-DOTA-NI-FAPI-04

[0151] Step 1: Synthesis of DOTA-NI-FAPI-04:

[0152] The synthetic route is as follows:

[0153] The specific steps include:

[0154] (1) Synthesis of Compound 6

[0155] Compound 4 (50.5 mg, 0.05 mmol) was dissolved in 2 mL of dichloromethane, 1 mL of diethylamine was added, and the mixture was stirred at room temperature for 2 hours. The reaction was detected by TLC, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light yellow oily intermediate. DOTA(tBu)3 (41 mg, 0.07 mmol) was dissolved in 2 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (27 mg, 0.07 mmol) and N,N'-diisopropylethylamine (13 mg, 0.1 mmol) were added under ice bath, and the mixture was stirred under ice bath. The mixture was stirred for 30 minutes to obtain a reaction solution. A light yellow oily intermediate (38 mg, 0.05 mmol) dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution and reacted at room temperature overnight. The mixture was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 10 / 1 / 0.1) to obtain yellow oily compound 6 (21.5 mg, 0.02 mmol) in a yield of 40%.

[0156] Structure confirmation of compound 6:

[0157] HRMS C 63 H 93 N 15 O 14 F2Na[M+Na] + Theoretical molecular weight 1344.6886, measured molecular weight 1344.6878;

[0158] 1 H NMR (600MHz, CD3OD) δ8.71(d,J=4.4Hz,1H),7.93(d,J=9.2Hz,2H),7.53(t,2H),7.41(dd,J=9.1,2.6Hz,1H),7.10(s,1H),5.09(dd,J=9.3,2.6Hz ,1H),4.45(t,J=7.2Hz,2H),4.30-4.18(m,5H),4.15-4.06(m,1H),3.76 (d,J=11.7Hz,1H),3.56-3.48(m,1H),3.40(d,J=5.6Hz,2H),3.27(dt,J= 3.2,1.6Hz,5H),3.20-3.14(m,2H),2.96-2.84(m,3H),2.76(dd,J=17.9 ,8.9Hz,3H),2.69-2.52(m,7H),2.45(d,J=27.2Hz,3H),2.23(t,J=19.9, 12.6Hz, 3H), 2.12-1.96 (m, 8H), 1.94 (d, J = 10.5Hz, 1H), 1.83 (dd, J = 13. 9,7.6Hz,1H),1.57-1.48(m,2H),1.47-1.38(m,27H),1.29-1.22(m,8H);

[0159] (2) Synthesis of compound DOTA-NI-FAPI-04

[0160] Compound 6 (21.5 mg, 0.02 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 2 hours. The solvent was removed by rotary evaporation under reduced pressure. The residue was dissolved in 2.1 mL of dimethyl sulfoxide and separated and purified using a semi-preparative chromatography column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-25 min, 5%-100% B; 25-26 min, 100%-5% B; 26-30 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain DOTA-NI-FAPI-04 as a white solid.

[0161] HRMS C51 H 69 N 15 O 14 F2[M+H] + Theoretical molecular weight 1154.5189, measured molecular weight 1154.5188;

[0162] Step 2: Ga-68 labeling of DOTA-NI-FAPI-04:

[0163] 68 Ga marked routes are as follows:

[0164] The marking method is as follows:

[0165] The DOTA-NI-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 23 μL of the precursor solution was placed in a 10 mL vial, and 72 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 95°C for 15 minutes, cooled to room temperature, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-DOTA-NI-FAPI-04;

[0166] As shown in FIG5 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-DOTA-NI-FAPI-04 shows that [ 68 The radiochemical purity of Ga]Ga-DOTA-NI-FAPI-04 is greater than 95%;

[0167] Example 6: 177 Preparation of Lu]Lu-DOTA-NI-FAPI-04

[0168] Step 1: Synthesis of DOTA-NI-FAPI-04:

[0169] The synthetic route and method are the same as those in step 1 of Example 5;

[0170] Step 2: Lu-177 labeling of DOTA-NI-FAPI-04:

[0171] The Lu-177 marked route is as follows:

[0172] The marking method is as follows:

[0173] Prepare a 1 μg / μL precursor solution of DOTA-NI-FAPI-04 obtained in step 1 with dimethyl sulfoxide. Take 24 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-DOTA-NI-FAPI-04;

[0174] As shown in FIG6, the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-NI-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-DOTA-NI-FAPI-04 was greater than 95%.

[0175] Example 7: 68 Preparation of [Ga]Ga-HBED-CC-NI-FAPI-04

[0176] Step 1: Synthesis of HBED-CC-NI-FAPI-04:

[0177] The synthetic route is as follows:

[0178] The specific steps include:

[0179] (1) Synthesis of compound 7

[0180] Compound 4 (50.5 mg, 0.05 mmol) was dissolved in 2 mL of dichloromethane, 1 mL of diethylamine was added, and the mixture was stirred at room temperature for 2 hours. The reaction was detected by TLC, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light yellow oily intermediate. 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu)2, 45 mg, 0.07 mmol) was dissolved in 2 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphonium was added under ice bath. Ester (27 mg, 0.07 mmol) and N,N'-diisopropylethylamine (13 mg, 0.1 mmol) were added to the reaction mixture and stirred in an ice bath for 30 minutes to obtain a reaction solution. A light yellow intermediate (38 mg, 0.05 mmol) dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution and reacted at room temperature overnight. The mixture was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain yellow oily compound 7 (21 mg, 0.02 mmol) in a yield of 40%.

[0181] Structure confirmation of compound 7:

[0182] HRMS C 69 H 90 N 13 O 16 F2[M+H] + Theoretical molecular weight 1394.6591, measured molecular weight 1394.6599;

[0183] 1H NMR (600MHz, CD3OD) δ8.70(t,J=5.1Hz,1H),7.92(dd,J=11.9,6.0Hz1H),7.52(t,J=4.2Hz ,1H),7.45-7.39(m,1H),7.08(d,J=4.9Hz,1H),6.97(dd,J=12.5,5.0Hz,1H),6.89-6.81( m,1H),6.66(t,J=9.0Hz,1H),5.08(t,J=11.8Hz,1H),4.45(dd,J=13.5,6.7Hz,2H),4.30- 4.23(m,3H),4.22-4.17(m,1H),4.14-4.05(m,1H),3.69(d,J=6.3Hz,3H),3.64(d,J=6.8Hz ,3H),3.60-3.50(m,2H),3.27(dt,J=3.2,1.6Hz,7H),3.26-3.23(m,3H),3.15(dt,J=13.9 ,6.6Hz,1H),2.81-2.71(m,6H),2.68(dd,J=15.3,8.3Hz,2H),2.55(dd,J=24.7,17.2Hz,4 H),2.45(dt,J=16.3,7.4Hz,3H),2.22-2.12(m,2H),2.07(dd,J=13.3,6.4Hz,2H),2.03-1 .92(m,3H),1.77-1.64(m,1H),1.60-1.48(m,1H),1.45-1.38(m,14H),1.33-1.22(m,15H);

[0184] (2) Synthesis of compound HBED-CC-NI-FAPI-04

[0185] Compound 7 (21 mg, 0.02 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 2 hours. The solvent was removed by rotary evaporation under reduced pressure. The residue was dissolved in 2.1 mL of dimethyl sulfoxide and separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-25 min, 5%-100% B; 25-26 min, 100%-5% B; 26-30 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain compound HBED-CC-NI-FAPI-04 as a white solid.

[0186] Structure confirmation of compound HBED-CC-NI-FAPI-04:

[0187] HRMS C 61 H 73 F2N 13 O 16 [M+H] + Theoretical molecular weight 1282.5339, measured molecular weight 1282.5331;

[0188] Step 2: Ga-68 labeling of HBED-CC-NI-FAPI-04:

[0189] The Ga-68 marked route is as follows:

[0190] The marking method is as follows:

[0191] The HBED-CC-NI-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 13 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 50°C for 10 minutes, cooled to room temperature, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-HBED-CC-NI-FAPI-04;

[0192] As shown in FIG7 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-HBED-CC-NI-FAPI-04 shows that [ 68 The radiochemical purity of [Ga]Ga-HBED-CC-NI-FAPI-04 was greater than 95%.

[0193] Example 8: 68 Preparation of Ga]Ga-NI-HBED-CC-FAPI-04

[0194] Step 1: Synthesis of NI-HBED-CC-FAPI-04:

[0195] The synthetic route is as follows:

[0196] The specific steps include:

[0197] (1) Synthesis of Compound 8

[0198] Compound 2 (130 mg, 0.48 mmol) was dissolved in 2 mL of trifluoroacetic acid solution, stirred at room temperature for 30 minutes, and evaporated under reduced pressure to remove the solvent to obtain a white solid intermediate; 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu)2, 309 mg, 0.48 mmol) was dissolved in 8 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (183 mg) was added under ice bath. , 0.48 mmol) and N,N'-diisopropylethylamine (187 mg, 1.45 mmol), stirred in an ice bath for 30 minutes to obtain a reaction solution, and a white solid intermediate dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution, and the reaction was allowed to proceed at room temperature for 5 hours. The resulting reaction solution was washed with ethyl acetate (20 mL×3) and saturated brine (20 mL×2), and the organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent, and purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to obtain compound 8 (55 mg, 0.07 mmol) as a light yellow oil; yield: 15%;

[0199] Structure confirmation of compound 8:

[0200] HRMS C 40 H 56 N6O 11 [M+H] + Theoretical molecular weight 797.4085, measured molecular weight 797.4081;

[0201] 1H NMR (400MHz, CD3OD) δ7.37(s,1H),7.08(d,J=1.0Hz,1H),7.01(dd,J=8.2,1.9Hz,2H),6.92(d,J=2.6Hz,2H),6.69(d ,J=2.8Hz,,1H),6.67(d,J=2.8Hz,1H),4.27(t,J=7.1Hz,2H),4.05(dd,1H),3.82(s,2H),3.75(s,2H),3.35(s,2H),3 .31(s,2H),3.26(dt,J=3.3,1.6Hz,2H),3.15(t,J=6.6Hz,2H),2.88(dd,J=7.8,3.9Hz,4H),2.76(dd,J=16.5,7.9Hz ,4H),2.47(t,J=7.7Hz,2H),2.41(t,J=7.5Hz,2H),1.96(d,J=0.5Hz,2H),1.95-1.87(m,2H),1.42(d,J=1.8Hz,18H);

[0202] (2) Synthesis of compound 9

[0203] Compound 8 (54.7 mg, 0.069 mmol) was dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (26 mg, 0.069 mmol) and N,N'-diisopropylethylamine (27 mg, 0.069 mmol) were added under ice bath, and stirred in ice bath for 30 minutes to obtain a reaction solution. (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1- [pyrrolidinyl]-2-oxoethyl]quinoline-4-carboxamide (CAS: 2374782-82-6, 33.6 mg, 0.069 mmol) was added to the above reaction solution and reacted at room temperature for 5 hours. The mixture was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 10 / 1 / 0.1) to obtain compound 9 (31.7 mg, 0.025 mmol) as a light yellow oil; yield: 36%;

[0204] HRMS C 64 H 82 N 12 O 143 F2[M+H] +Theoretical molecular weight 1265.617, measured molecular weight 1265.6169;

[0205] 1 H NMR (400MHz, CD3OD) δ8.69(d,J=4.5Hz,1H),7.94-7.87(m,2H),7.51(d,J=4.4Hz,1H),7.40(d,J=2.7Hz,1H),7.39-7.35(m, 1H),7.10-7.04(m,1H),7.02-6.94(m,2H),6.88(dd,J=11.9,1.7Hz,2H),6.67(dd,J=12.9,8.2Hz,2H),5.09(dd,J=9.3,3.0H z,1H),4.32-4.17(m,7H),3.74(s,2H),3.68(s,2H),3.59(s,2H),3.47(s,2H),3.29-3.24(m,7H),3.13(t,J=6.5Hz,2H),2.8 2-2.68(m,11H),2.63-2.54(m,4H),2.48(s,2H),2.39(t,J=7.5Hz,2H),2.11-2.01(m,2H),1.94-1.85(m,2H),1.41(s,18H);

[0206] (3) Synthesis of compound NI-HBED-CC-FAPI-04

[0207] Compound 9 (31.7 mg, 0.025 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was removed by rotary evaporation under reduced pressure. The residue was dissolved in dimethyl sulfoxide and separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-18 min, 5%-59% B; 18-20 min, 59% B; 20-21 min, 59%-5% B; 21-25 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain NI-HBED-CC-FAPI-04 as a white solid.

[0208] Structure confirmation of compound NI-HBED-CC-FAPI-04:

[0209] HRMS C 56 H 67 N 12 O 13 F2[M+H] +Theoretical molecular weight 1153.4913, measured molecular weight 1153.4912;

[0210] Step 2: Ga-68 labeling of NI-HBED-CC-FAPI-04:

[0211] The Ga-68 marked route is as follows:

[0212] The marking method is as follows:

[0213] The NI-HBED-CC-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 12 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 50°C for 10 minutes, cooled to room temperature, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-NI-HBED-CC-FAPI-04;

[0214] As shown in FIG8 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-NI-HBED-CC-FAPI-04 shows that [ 68 The radiochemical purity of Ga]Ga-NI-HBED-CC-FAPI-04 was greater than 95%.

[0215] Example 9: 68 Preparation of Ga]Ga-NI-HBED-CC-FAPI-02

[0216] Step 1: Synthesis of NI-HBED-CC-FAPI-02:

[0217] The synthetic route is as follows:

[0218] The specific steps include:

[0219] (1) Synthesis of compound 10

[0220] (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2370952-97-7, 35 mg, 0.064 mmol) was dissolved in 3 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was removed by rotary evaporation under reduced pressure to obtain an orange-red intermediate. Compound 8 was dissolved in 3 mL of ultra-dry N,N-dimethylformamide and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (49 mg, 0.13 mmol) and N,N'-diisopropylamine were added under ice-bath conditions. The mixture was stirred in an ice bath for 30 minutes to obtain a reaction solution. The orange-red intermediate dissolved in 2 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution and reacted at room temperature for 5 hours. The reaction solution was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The product was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain compound 10 (38.4 mg, 0.031 mmol) as a light yellow oil in a yield of 48%.

[0221] Structure confirmation of compound 10:

[0222] HRMS C 64 H 85 N 12 O 13 [M+H] + Theoretical molecular weight 1229.6353, measured molecular weight 1229.6356;

[0223] (2) Synthesis of compound NI-HBED-CC-FAPI-02

[0224] Compound 10 (38.4 mg, 0.031 mmol) was dissolved in 2 mL of trifluoroacetic acid and stirred at room temperature for 1 hour. The solvent was removed by rotary evaporation under reduced pressure, and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-25 min, 5%-80% B; 25-26 min, 80%-5% B; 26-30 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain NI-HBED-CC-FAPI-02 as a white solid.

[0225] Structure confirmation of compound NI-HBED-CC-FAPI-02:

[0226] HRMS C 56 H 68 N 12 O 13 [M+H] + Theoretical molecular weight 1117.5101, measured molecular weight 1117.5104;

[0227] Step 2: Ga-68 labeling of NI-HBED-CC-FAPI-02:

[0228] The Ga-68 marked route is as follows:

[0229] The marking method is as follows:

[0230] The NI-HBED-CC-FAPI-02 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 11 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 50°C for 10 minutes, cooled to room temperature, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-NI-HBED-CC-FAPI-02;

[0231] As shown in FIG9 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-NI-HBED-CC-FAPI-02 shows that [ 68 The radiochemical purity of Ga]Ga-NI-HBED-CC-FAPI-02 was greater than 95%.

[0232] Example 10: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-FAPI-02

[0233] Step 1: Synthesis of DOTA-HBED-CC-FAPI-02:

[0234] The synthetic route is as follows:

[0235] The specific steps include:

[0236] (1) Synthesis of compound 11

[0237] In a 100 mL round-bottom flask, 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)acetic acid (DOTA(tBu)3, 1 g, 1.75 mmol) was dissolved in 15 mL of ultra-dry N,N-dimethylformamide. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (666 mg, 1.75 mmol) and N,N'-diisopropylethylamine (679 mg, 5.25 mmol) were added under ice bath. The mixture was stirred for 30 minutes while keeping the ice bath. The reaction mixture was added with N-Cbz-1,2-diaminoethane (408 mg, 2.1 mmol) dissolved in 5 mL of ultra-dry N,N-dimethylformamide. The mixture was reacted at room temperature for 5 hours, and the mixture was washed with ethyl acetate (20 mL × 3) and saturated brine (20 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, and the filtrate was evaporated under reduced pressure to remove the solvent. The mixture was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 20 / 1 / 0.1) to obtain compound 11 (1.02 g, 1.36 mmol) as a yellow solid in a yield of 78%.

[0238] Structure confirmation of compound 11:

[0239] HRMS C 38 H 65 N6O9[M+H] + Theoretical molecular weight 749.4807, measured molecular weight 749.4802;

[0240] 1 H NMR (400MHz, CDCl3) δ7.34(d,J=1.9Hz,4H),7.32(s,1H),6.58(t,J=4.7Hz,1H),5.07(s,2H),3.33(s,8H),2.80(s,4H),1.45(s,16H),1.42(s,27H);

[0241] (2) Synthesis of compound 12

[0242] In a 100 mL round-bottom flask, compound 11 (1.02 g, 1.36 mmol) was dissolved in a mixed solution of 20 mL tetrahydrofuran and 10 mL methanol. 10% Pd / C (150 mg, 0.14 mmol) was added, and the mixture was stirred at room temperature under a hydrogen atmosphere overnight. The resulting reaction solution was filtered through celite, and the filtrate was evaporated under reduced pressure to remove the solvent to obtain compound 12 (852 mg, 1.38 mmol) as a white solid with a yield of >99%. The product was carried on to the next step without purification.

[0243] (3) Synthesis of Compound 13

[0244] In a 100 mL round-bottom flask, 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu)2, 500 mg, 0.78 mmol) was dissolved in 20 mL of ultra-dry N,N-dimethylformamide, and N,N'-diisopropylethylamine (300 mg, 2.3 mmol) was added under ice-bath conditions. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate) dissolved in 10 mL of ultra-dry N,N-dimethylformamide was added dropwise. The reaction mixture was stirred in an ice bath for 30 minutes to obtain a reaction solution. Compound 12 (479 mg, 0.78 mmol) dissolved in 10 mL of ultra-dry N,N-dimethylformamide was added to the above reaction solution, and the mixture was reacted at room temperature for 5 hours. The mixture was evaporated under reduced pressure to remove most of the N,N-dimethylformamide. The residue was washed with ethyl acetate (20 mL × 3) and saturated brine (20 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain compound 13 (234 mg, 0.19 mmol) as a colorless solid in a yield of 24%.

[0245] Structure confirmation of compound 13:

[0246] HRMS C 64 H 105 N8O 16 [M+H] + Theoretical molecular weight 1241.7643, measured molecular weight 1241.7652;

[0247] 1 H NMR (400MHz, CDCl3) δ9.22(s,1H),6.97(d,J=7.9Hz,2H),6.84(s,1H),6.79(s,1H),6.70(dd,J=12.0,8.3Hz,2H),6.60(s,1H),6.45(s, 1H),3.65(s,4H),3.23(s,10H),3.12-2.63(m,16H),2.63-2.52(m,4H),2.51-2.38(m,4H),2.09(d,J=44.2Hz,7H),1.49-1.33(m,45H);

[0248] (4) Synthesis of Compound 14

[0249] In a 50 mL round-bottom flask, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2370952-97-7, 21 mg, 0.038 mmol) was dissolved in 3 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was then dried to give an orange-red intermediate. In a 50 mL round-bottom flask, compound 13 (36 mg, 0.029 mmol) was dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and N,N'-diisopropylethylamine (15 mg, 0.12 mmol) and 2-(7-azobenzotriazole)-N,N, N', N'-tetramethyluronium hexafluorophosphate (14 mg, 0.037 mmol) was stirred in an ice bath for 30 minutes to obtain a reaction solution. The above orange-red intermediate was dissolved in 5 mL of ultra-dry N, N-dimethylformamide and added to the above reaction solution. The reaction was carried out at room temperature for 5 hours. The majority of N, N-dimethylformamide was removed by pressure rotary evaporation. The residue was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by vacuum rotary evaporation. The residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain compound 14 (15 mg, 0.009 mmol) as a light yellow oil in a yield of 31%.

[0250] Structure confirmation of compound 14:

[0251] HRMS C 88 H 133 N 14 O 18 [M+H] + Theoretical molecular weight 1673.9916, measured molecular weight 1673.9922;

[0252] (5) Synthesis of compound DOTA-HBED-CC-FAPI-02

[0253] In a 25 mL round-bottom flask, compound 14 (15 mg, 0.009 mmol) was dissolved in 4 mL of trifluoroacetic acid and stirred at room temperature for 2.5 hours. The solvent was removed by rotary evaporation under reduced pressure, and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-15 min, 5%-59% B; 15-16 min, 59%-5% B; 16-20 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain DOTA-HBED-CC-FAPI-02 as a white solid with a purity of >95% as determined by LC-MS.

[0254] Structure confirmation of compound DOTA-HBED-CC-FAPI-02:

[0255] HRMS C 68 H 93 N 14 O 18 [M+H] + Theoretical molecular weight 1393.6786, measured molecular weight 1393.6784;

[0256] Step 2: Ga-68 labeling of DOTA-HBED-CC-FAPI-02:

[0257] The Ga-68 marked route is as follows:

[0258] The marking method is as follows:

[0259] The DOTA-HBED-CC-FAPI-02 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 14 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the radioligand sodium acetate mixed solution. After mixing evenly, react at room temperature for 10 minutes to obtain DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02;

[0260] As shown in FIG10 , the DOTA-[ 68The radioactive HPLC spectrum of the labeled reaction solution of DOTA-[Ga]Ga-HBED-CC-FAPI-02 shows that 68 The radiochemical purity of [Ga]Ga-HBED-CC-FAPI-02 was greater than 95%.

[0261] Example 11: 177 Preparation of Lu]Lu-DOTA-HBED-CC-FAPI-02

[0262] Step 1: Synthesis of DOTA-HBED-CC-FAPI-02:

[0263] The synthetic route and method are the same as those in step 1 in Example 10;

[0264] Step 2: Lu-177 labeling of DOTA-HBED-CC-FAPI-02:

[0265] The Lu-177 marked route is as follows:

[0266] The marking method is as follows:

[0267] Prepare a 1 μg / μL precursor solution of DOTA-HBED-CC-FAPI-02 obtained in step 1 with dimethyl sulfoxide. Take 28 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02;

[0268] As shown in FIG11 , the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-02 shows that [ 177 The radiochemical purity of Lu]Lu-DOTA-HBED-CC-FAPI-02 was greater than 95%.

[0269] Example 12: DOTA-[ 68 Preparation of [Ga]Ga-HBED-CC-FAPI-04)

[0270] Step 1: Synthesis of DOTA-HBED-CC-FAPI-04:

[0271] The synthetic route is as follows:

[0272] (1) Synthesis of compound 15

[0273] In a 50 mL round-bottom flask, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2374782-82-6, 25 mg, 0.043 mmol) was dissolved in 3 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was then dried to give a yellow intermediate. In a 50 mL round-bottom flask, compound 13 (31 mg, 0.025 mmol) was dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and N,N'-diisopropylethylamine (15 mg, 0.12 mmol) and 2-(7-azobenzotriazole) were added under ice-bath conditions. )-N,N,N',N'-tetramethyluronium hexafluorophosphate (15 mg, 0.039 mmol), stirred in an ice bath for 30 minutes to obtain a reaction solution. The above yellow intermediate was dissolved in 5 mL of ultra-dry N,N-dimethylformamide and added to the above reaction solution. The reaction was carried out at room temperature for 5 hours. The majority of N,N-dimethylformamide was removed by pressure rotary evaporation. The residue was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by reduced pressure rotary evaporation. The residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain light yellow oily compound 15 (29 mg, 0.017 mmol) in a yield of 68%.

[0274] Structure confirmation of compound 15:

[0275] HRMS C 88 H 132 N 14 O 18 F2[M+2H] 2+ Theoretical molecular weight 855.49, measured molecular weight 855.4894;

[0276] (2) Synthesis of compound DOTA-HBED-CC-FAPI-04

[0277] In a 25 mL round-bottom flask, compound 15 (29 mg, 0.017 mmol) was dissolved in 4 mL of trifluoroacetic acid and stirred at room temperature for 2.5 hours. The solvent was removed by rotary evaporation under reduced pressure, and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-15 min, 5%-50% B; 15-16 min, 50%-5% B; 16-20 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target component was lyophilized to obtain DOTA-HBED-CC-FAPI-04 as a white solid with a purity of >95% as determined by LC-MS.

[0278] Structure confirmation of compound DOTA-HBED-CC-FAPI-04:

[0279] HRMS C 68 H 91 N 14 O 18 F2[M+H] + Theoretical molecular weight 1429.6598, measured molecular weight 1429.6592;

[0280] Step 2: Ga-68 labeling of DOTA-HBED-CC-FAPI-04:

[0281] The Ga-68 marked route is as follows:

[0282] The marking method is as follows:

[0283] The DOTA-HBED-CC-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 14 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the radioligand sodium acetate mixed solution, mix well, and react at room temperature for 10 minutes to obtain DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04;

[0284] As shown in FIG12 , the DOTA-[ 68The radioactive HPLC spectrum of the labeled reaction solution of DOTA-[Ga]Ga-HBED-CC-FAPI-04 was shown in the spectrum. 68 The radiochemical purity of [Ga]Ga-HBED-CC-FAPI-04 was greater than 95%.

[0285] Example 13: 177 Preparation of Lu]Lu-DOTA-HBED-CC-FAPI-04

[0286] Step 1: Synthesis of DOTA-HBED-CC-FAPI-04:

[0287] The synthetic route and method are the same as those in step 1 in Example 12;

[0288] Step 2: Lu-177 labeling of DOTA-HBED-CC-FAPI-04:

[0289] The Lu-177 marked route is as follows:

[0290] The marking method is as follows:

[0291] Prepare a 1 μg / μL precursor solution of DOTA-HBED-CC-FAPI-04 obtained in step 1 with dimethyl sulfoxide. Take 29 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04;

[0292] As shown in FIG13, the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-DOTA-HBED-CC-FAPI-04 was greater than 95%.

[0293] Example 14: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-PEG3-FAPI-02

[0294] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-02:

[0295] The synthetic route is as follows:

[0296] The specific steps include:

[0297] (1) Synthesis of compound 16

[0298] In a 50 mL round-bottom flask, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2370952-97-7, 212 mg, 0.39 mmol) was dissolved in 3 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was spin-dried to obtain an orange-red substance, which was dissolved in 10 mL of anhydrous acetonitrile and anhydrous potassium carbonate (216 mg, 1.56 mmol) was added. The mixture was stirred at room temperature for 30 minutes. The mixture was stirred for 30 minutes, and tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamate (167 mg, 0.47 mmol) was added. The mixed solution was refluxed at 80° C. overnight, and potassium carbonate was removed by filtration through celite. The filtrate was evaporated under reduced pressure to remove the solvent, and the residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to give compound 16 (191 mg, 0.26 mmol) as a light yellow oil in a yield of 67%.

[0299] Structure confirmation of compound 16:

[0300] HRMS C 37 H 56 N7O8[M+H] + Theoretical molecular weight 726.419, measured molecular weight 726.4179;

[0301] 1 H NMR(400MHz, CDCl3) δ8.66(d,J=4.4Hz,1H),7.92(dd,J=9.2,4.3Hz,1H),7.62(d,J=2.3Hz,1 H),7.51(s,1H),7.41(d,J=4.4Hz,1H),7.30(dd,J=9.2,2.6Hz,1H),4.74(dd,J=23.1,6.2Hz ,1H),4.15-4.05(m,2H),3.68-3.51(m,12H),3.51-3.43(m,3H),3.36(s,1H),3.25(d,J=3.4 Hz,2H),2.62(s,12H),2.21(tdd,J=17.5,10.5,7.9Hz,4H),2.08-1.95(m,2H),1.37(s,9H);

[0302] (2) Synthesis of Compound 17

[0303] In a 50 mL round-bottom flask, compound 16 (32 mg, 0.044 mmol) was dissolved in 3 mL of trifluoroacetic acid, stirred at room temperature for 30 minutes, and the solvent was spin-dried to obtain an orange-yellow intermediate. In a 50 mL round-bottom flask, compound 13 (37 mg, 0.03 mmol) was dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and N,N'-diisopropylethylamine (15 mg, 0.12 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (17 mg, 0.045 mmol) were added under ice bath conditions, and stirred in an ice bath for 30 minutes to obtain a reaction solution. The above orange-yellow intermediate was dissolved in 5 mL of ultra-dry N,N-dimethylformamide. The reaction mixture was added with formamide and the reaction mixture was reacted at room temperature for 5 hours. Most of the N,N-dimethylformamide was removed by pressure rotary evaporation. The residue was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by vacuum rotary evaporation. The residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to give compound 17 (31 mg, 0.017 mmol) as a light yellow oil in a yield of 57%.

[0304] Structure confirmation of compound 17:

[0305] HRMS C 96 H 150 N 15 O 21 Na[M+H+Na] 2+ Theoretical molecular weight 936.0508, measured molecular weight 936.0511;

[0306] (3) Synthesis of compound DOTA-HBED-CC-PEG3-FAPI-02

[0307] In a 25 mL round-bottom flask, compound 17 (31 mg, 0.017 mmol) was dissolved in 4 mL of trifluoroacetic acid and stirred at room temperature for 2.5 hours. The solvent was removed by rotary evaporation under reduced pressure, and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-15 min, 5%-50% B; 15-16 min, 50%-5% B; 16-20 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target fraction was lyophilized to obtain DOTA-HBED-CC-PEG3-FAPI-02 as a white solid with a purity of >95% as determined by LC-MS.

[0308] The structure of the compound DOTA-HBED-CC-PEG3-FAPI-02 was confirmed:

[0309] HRMS C 76 H 110 N 15 O 21 [M+H] + Theoretical molecular weight 1568.7995, measured molecular weight 1568.7989;

[0310] Step 2: Ga-68 labeling of DOTA-HBED-CC-PEG3-FAPI-02:

[0311] The Ga-68 marked route is as follows:

[0312] The marking method is as follows:

[0313] The DOTA-HBED-CC-PEG3-FAPI-02 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 16 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the radioligand sodium acetate mixed solution, mix well, and react at room temperature for 10 minutes to obtain DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02.

[0314] As shown in FIG14 , the DOTA-[ 68 The radioactive HPLC spectrum of the labeled reaction solution of DOTA-[Ga]Ga-HBED-CC-PEG3-FAPI-02 was shown in the spectrum. 68 The radiochemical purity of [Ga]Ga-HBED-CC-PEG3-FAPI-02 was greater than 95%.

[0315] Example 15: 177 Preparation of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02

[0316] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-02:

[0317] The synthetic route and method are the same as those in step 1 in Example 14;

[0318] Step 2: Lu-177 labeling of DOTA-HBED-CC-PEG3-FAPI-02:

[0319] The Lu-177 marked route is as follows:

[0320] The marking method is as follows:

[0321] Prepare a 1 μg / μL precursor solution of DOTA-HBED-CC-PEG3-FAPI-02 obtained in step 1 with dimethyl sulfoxide. Take 31 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02.

[0322] As shown in FIG15 , the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02 shows that [ 177 The radiochemical purity of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02 was greater than 95%.

[0323] Example 16: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-PEG3-FAPI-04

[0324] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-04:

[0325] The synthetic route is as follows:

[0326] The specific steps include:

[0327] (1) Synthesis of Compound 18

[0328] In a 50 mL round-bottom flask, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide (CAS: 2374782-82-6, 105 mg, 0.18 mmol) was dissolved in 3 mL of trifluoroacetic acid and stirred at room temperature for 30 minutes. The solvent was dried to obtain a yellow substance, which was dissolved in 10 mL of anhydrous acetonitrile and anhydrous potassium carbonate (106 mg, 0.77 mm Hg) was added. ol), stirred at room temperature for 30 minutes, and tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamate (82 mg, 0.23 mmol) was added. The mixed solution was refluxed at 80°C overnight, and potassium carbonate was removed by filtration through celite. The filtrate was evaporated under reduced pressure to remove the solvent, and the residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 10 / 1 / 0.1) to give compound 18 (131 mg, 0.17 mmol) as a light yellow oil in a yield of 94%;

[0329] Structure confirmation of compound 18:

[0330] HRMS C 37 H 54 N7O8F2[M+H] + Theoretical molecular weight 762.3996, measured molecular weight 762.3992;

[0331] 1 H NMR (600MHz, CDCl3) δ8.57(d,J=28.1Hz,1H),7.85(d,J=5.6Hz,2H),7.56(d,J=26.6Hz,1H),7.33(s,1H),5.26(s,1H),4.93(s,1H),4.22(t,J=21.4Hz ,1H),4.03(d,J=35.2Hz,4H),3.95-3.78(m,2H),3.65-3.33(m,13H),3.20( d,J=21.6Hz,2H),2.61(d,J=43.8Hz,13H),1.99(s,2H),1.44-1.26(m,9H);

[0332] (2) Synthesis of compound 19

[0333] In a 50 mL round-bottom flask, compound 18 (33 mg, 0.043 mmol) was dissolved in 3 mL of trifluoroacetic acid, stirred at room temperature for 30 minutes, and the solvent was spin-dried to obtain a light yellow intermediate; in a 50 mL round-bottom flask, compound 13 (32 mg, 0.026 mmol) was dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and N,N'-diisopropylethylamine (13 mg, 0.10 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (13 mg, 0.034 mmol) were added under ice bath conditions, and stirred under ice bath conditions. The mixture was stirred for 30 minutes to obtain a reaction solution. The above light yellow intermediate was dissolved in 5 mL of ultra-dry N,N-dimethylformamide and added to the above reaction solution. The mixture was reacted at room temperature for 5 hours. The mixture was evaporated under reduced pressure to remove most of the N,N-dimethylformamide. The residue was washed with ethyl acetate (10 mL × 3) and saturated brine (10 mL × 2). The organic phase was collected and dried over anhydrous sodium sulfate. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain compound 19 (28 mg, 0.015 mmol) as a light yellow oil in a yield of 58%.

[0334] Structure confirmation of compound 19:

[0335] HRMS C 96 H 149 N 15 O 21 F2[M+2H] 2+ Theoretical molecular weight 943.0504, measured molecular weight 943.0520;

[0336] (3) Synthesis of compound DOTA-HBED-CC-PEG3-FAPI-04

[0337] In a 25 mL round-bottom flask, compound 19 (28 mg, 0.015 mmol) was dissolved in 4 mL of trifluoroacetic acid and stirred at room temperature for 2.5 hours. The solvent was removed by rotary evaporation under reduced pressure, and the residue was dissolved in dimethyl sulfoxide. The mixture was separated and purified using a semi-preparative column (Eclipse XDB-C18, 5 μm, 9.4×250 mm) (phase A: 0.1% trifluoroacetic acid-water, phase B: 0.1% trifluoroacetic acid-acetonitrile; gradient: 0-15 min, 5%-50% B; 15-16 min, 50%-5% B; 16-20 min, 5% B; flow rate: 4 mL / min, UV: 280 nm). The target fraction was lyophilized to obtain DOTA-HBED-CC-PEG3-FAPI-04 as a white solid with a purity of >95% as determined by LC-MS.

[0338] The structure of the compound DOTA-HBED-CC-PEG3-FAPI-04 was confirmed:

[0339] HRMS C 76 H 108 N 15 O 21 F2[M+H] + Theoretical molecular weight 1604.7806, measured molecular weight 1604.7778;

[0340] Step 2: Ga-68 labeling of DOTA-HBED-CC-PEG3-FAPI-04:

[0341] The Ga-68 marked route is as follows:

[0342] The marking method is as follows:

[0343] The DOTA-HBED-CC-PEG3-FAPI-04 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 16 μL of the precursor solution was placed in a 10 mL vial, and 135 μL of 3 M sodium acetate solution was added to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba) was rinsed with 6 mL of 0.6 M high-purity hydrochloric acid solution to obtain [ 68 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the radioligand sodium acetate mixed solution, mix well, and react at room temperature for 10 minutes to obtain DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04;

[0344] As shown in FIG16 , the DOTA-[ 68 The radioactive HPLC spectrum of the labeled reaction solution of DOTA-[Ga]Ga-HBED-CC-PEG3-FAPI-04 was shown in the figure. 68 The radiochemical purity of [Ga]Ga-HBED-CC-PEG3-FAPI-04 was greater than 95%.

[0345] Example 17: 177 Preparation of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04

[0346] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-04:

[0347] The synthetic route and method are the same as those in step 1 in Example 16;

[0348] Step 2: Lu-177 labeling of DOTA-HBED-CC-PEG3-FAPI-04:

[0349] The Lu-177 marked route is as follows:

[0350] The marking method is as follows:

[0351] Prepare a 1 μg / μL precursor solution of DOTA-HBED-CC-PEG3-FAPI-04 obtained in step 1 with dimethyl sulfoxide. Take 32 μL of the precursor solution and place it in a 10 mL vial. Add 12 μL of 3 M sodium acetate solution and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95 ° C for 20 minutes, and its radiochemical purity was determined by high performance liquid chromatography with a radioactivity detector to obtain [ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04.

[0352] As shown in FIG17 , the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 was greater than 95%.

[0353] Application Example 1

[0354] DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 In vitro HT1080-FAP cell uptake of [Ga]Ga-DOTA-FAPI-04:

[0355] HT1080-FAP cells were prepared to a density of 1.25 × 10 6 400 μL of cell suspension was inoculated into 6-well plates and cultured for 24 hours. 37-111 KBq of DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 Ga]Ga-DOTA-FAPI-04 solution was added and incubated in a constant temperature box at 37°C. At 10, 30, 60, 90 and 120 minutes of incubation in each group, PBS solution was used to terminate its uptake, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube to determine the radioactivity count; for the blocking experiment, excess unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drug. After incubation for 60 minutes, PBS solution was used to terminate its uptake, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0356] As shown in FIG18 , the uptake of DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 Ga]Ga-DOTA-FAPI-04 uptake-time curve (n=3); as shown in FIG19 , the in vitro HT1080-FAP cell uptake of DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 60 min uptake and blocking results of Ga]Ga-DOTA-FAPI-04 (n=3).

[0357] The results of in vitro cell uptake experiments showed that DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 The uptake of Ga]Ga-HBED-CC-PEG3-FAPI-04 in FAP-positive HT1080-FAP cells gradually increased with the extension of incubation time, and the uptake value of the tested drug was higher than that of the positive control at any time point monitored. 68 The uptake of all drugs could be blocked by excess DOTA-FAPI-04, indicating that all drugs specifically targeted FAP.

[0358] Application Example 2

[0359] [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177 In vitro HT1080-FAP cell uptake of Lu]Lu-DOTA-FAPI-04

[0360] HT1080-FAP cells were prepared to a density of 1.25 × 10 6 400 μL of the cell suspension was inoculated into 6-well plates and cultured for 24 hours. 81 KBq of [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177Lu]Lu-DOTA-FAPI-04 solution and incubated in a constant temperature box at 37 ° C. At 10, 30, 60, 90 and 120 minutes of incubation in each group, PBS solution was used to terminate its uptake, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube to determine the radioactivity count; for the blocking experiment, excess unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drug. After incubation for 60 minutes, PBS solution was used to terminate its uptake, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0361] As shown in FIG20 , the in vitro HT1080-FAP cell uptake in Example 2 of the present invention is shown. 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177 Lu]Lu-DOTA-FAPI-04 uptake-time curve (n=3); as shown in FIG21, the in vitro HT1080-FAP cell uptake of [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177 60-min uptake and blocking results of Lu]Lu-DOTA-FAPI-04 (n=3).

[0362] Application Example 3

[0363] [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 In vitro HT1080-FAP cell uptake of Lu]Lu-DOTA-FAPI-04

[0364] HT1080-FAP cells were prepared to a density of 1.25 × 10 6 400 μL of the cell suspension was inoculated into 6-well plates and cultured for 48 hours. 18.5 KBq of [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 Lu]Lu-DOTA-FAPI-04 solution was incubated at 37°C for 10, 30, 60, 90 and 120 minutes in each group, and its uptake was terminated with PBS solution. The cells were lysed with 1M NaOH, and the cell lysate was aspirated with filter paper and plugged into a plastic test tube for determination of radioactivity count. For blocking experiments, excess unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drug. After 60 minutes of incubation, its uptake was terminated with PBS solution, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube for determination of radioactivity count.

[0365] As shown in FIG22 , the in vitro HT1080-FAP cell uptake in Example 3 of the present invention is shown. 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 Lu]Lu-DOTA-FAPI-04 uptake-time curve (n=3); as shown in FIG23, the in vitro HT1080-FAP cell uptake of [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 60-min uptake and blocking results of Lu]Lu-DOTA-FAPI-04 (n=3).

[0366] The results of in vitro cell uptake experiments showed that [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177The uptake of Lu]Lu-DOTA-FAPI-04 in FAP-positive HT1080-FAP cells gradually increased with the extension of incubation time, and the uptake of all drugs could be blocked by excess DOTA-FAPI-04, indicating that all drugs specifically targeted FAP; at all monitored time points, [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 The uptake values ​​of Lu]Lu-DOTA-NI-FAPI-04 were all the same as [ 177 Lu]Lu-DOTA-FAPI-04 is similar or slightly higher than [ 177 Lu]Lu-DOTA-FAPI-04.

[0367] Application Example 4

[0368] [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 In vitro HT1080-FAP cell uptake of [Ga]Ga-DOTA-FAPI-04:

[0369] HT1080-FAP cells were prepared to 4 × 10 5 500 μL of the cell suspension was inoculated into 6-well plates and cultured for 60 hours. 222 KBq of [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 Ga]Ga-DOTA-FAPI-04 solution was incubated at 37°C for 10, 30, 60, 90 and 120 minutes in each group, and its uptake was terminated with PBS solution. The cells were lysed with 1M NaOH, and the cell lysate was aspirated with filter paper and plugged into a plastic test tube for determination of radioactivity count. For blocking experiments, excess unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drugs. After 60 minutes of incubation, its uptake was terminated with PBS solution, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and plugged into a plastic test tube for determination of radioactivity count.

[0370] As shown in FIG24 , the in vitro HT1080-FAP cell uptake in Example 4 of the present invention is shown. 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 Ga]Ga-DOTA-FAPI-04 uptake-time curve (n=3); as shown in FIG25, the in vitro HT1080-FAP cell uptake of [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 60 min uptake and blocking results of Ga]Ga-DOTA-FAPI-04 (n=3).

[0371] The results of in vitro cell uptake experiments showed that [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 The uptake of [Ga]Ga-NI-HBED-CC-FAPI-02 in FAP-positive HT1080-FAP cells gradually increased with the extension of incubation time, and the uptake of all drugs could be blocked by excess DOTA-FAPI-04, indicating that all drugs specifically targeted FAP; at all monitored time points, [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-02 and [ 68 The uptake values ​​of Ga]Ga-DOTA-FAPI-04 were higher than those of [ 68 Ga]Ga-DOTA-FAPI-04.

[0372] Application Example 5

[0373] [ 68 Ga]Ga-AAZTA-FAPI-04, [ 68 Ga]Ga-AAZTA-NI-FAPI-04, [ 68Ga]Ga-DOTA-NI-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 In vivo PET imaging of U87MG tumor-bearing mice with Ga]Ga-DOTA-FAPI-04.

[0374] In U87MG tumor-bearing mice, 5.55 MBq of [ 68 Ga]Ga-AAZTA-FAPI-04, [ 68 Ga]Ga-AAZTA-NI-FAPI-04, [ 68 Ga]Ga-DOTA-NI-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 or [ 68 Ga]Ga-DOTA-FAPI-04 (positive control), and then small animal PET / CT imaging was performed under isoflurane anesthesia at 10, 30, 60, 90, and 120 minutes after administration. For the blocking group, each drug was mixed with an excess of DOTA-FAPI-04 and then injected into the tail vein of tumor-bearing mice, and imaging was performed at 60 minutes.

[0375] As shown in FIG26, in the application example 4 of the present invention, [ 68 Ga]Ga-AAZTA-FAPI-04, [ 68 Ga]Ga-AAZTA-NI-FAPI-04, [ 68 Ga]Ga-DOTA-NI-FAPI-04, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04, DOTA-[ 68Ga]Ga-HBED-CC-PEG3-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-PEG3-FAPI-04 and [ 68 In vivo PET imaging results of U87MG tumor-bearing mice using Ga]Ga-DOTA-FAPI-04.

[0376] In vivo PET / CT imaging of mice bearing U87MG tumors showed that all compounds were taken up in the tumor site, and blocking experiments showed that drug uptake in the tumor could be blocked by excessive DOTA-FAPI-04, indicating that the drug specifically targets FAP. In addition, the tumor uptake and retention of all tested drugs were higher than those of the positive control. 68 Ga]Ga-DOTA-FAPI-04, and AAZTA-FAPI-04, AAZTA-NI-FAPI-04, DOTA-NI-FAPI-04, DOTA-HBED-CC-FAPI-02, DOTA-HBED-CC-FAPI-04, DOTA-HBED-CC-PEG3-FAPI-02, and DOTA-HBED-CC-PEG3-FAPI-04 can all be labeled with therapeutic radionuclides such as Lu-177, and are expected to become new FAP targeted radiodiagnosis and treatment integrated drugs.

[0377] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A radioactively labeled FAPI complex, the general structural formula thereof is as follows: Among them, R is a nitroimidazole group or -COOH, and the nitroimidazole group is selected from any one of the following: X is a chelating group or a chelating structure that chelates a radionuclide, and is selected from any one of the following: Among them, M includes but is not limited to 67 Ga 3+ 、 68 Ga 3+ 、[Al 18 F] 2+ 、 64 Cu 2+ 、 67 Cu 2+ 、 111 In 3+ 、 177 Lu 3+ 、 86 Y 3+ 、 90 Y 3+ 、 44 Sc 3+ 、 47 Sc 3+ 、 225 Ac 3+ 、 212 Pb 2+ 、 203 Pb 2+ 、 213 Bi 3+ or 212 Bi 3+ ; L 1 and L 2 is a linking group, wherein L 1 is selected from any one of the following: wherein, n is an integer from 0 to 6; L 2 Selected from any one of the following: A is a regulatory atom of the FAP targeting group, and A is either an H atom or an F atom.

2. The radiolabeled FAPI complex according to claim 1 has the following specific structure:

3. Preparation of a radioactively labeled FAPI complex, the steps are as follows: (1) Nitroimidazole and anhydrous potassium carbonate are dissolved in ultra-dry N,N-dimethylformamide, stirred at room temperature, N-(3-bromopropyl)carbamic acid tert-butyl ester is added, after the reaction is completed, suction filtration is carried out, extraction is carried out, the organic phase is dried and purified to obtain a yellowish-green oily compound, and deprotection is carried out with trifluoroacetic acid to obtain an intermediate 1 in the form of a white solid; (2) (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid is dissolved in ultra-dry N,N-dimethylformamide, under ice bath, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N’-diisopropylethylamine are added, stirred, and the white solid intermediate 1 obtained in step (1) is added. After the reaction is completed, extraction is carried out, the organic phase is dried and purified to obtain a pale yellow solid, deprotection is carried out with trifluoroacetic acid to obtain a yellow oil. The yellow oil is dissolved in N,N-dimethylformamide, 2-(7-azabenzotriazol)-N,N,N',N'-tetramethylurea hexafluorophosphate and N,N’-diisopropylethylamine are added under ice bath, stirred, and FAPI after deprotection of the Boc group with trifluoroacetic acid is added (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)aminocarbonyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)aminocarbonyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester). After the reaction is completed, extraction is carried out, the organic phase is dried and purified to obtain a pale yellow oil, and deprotection is carried out with diethylamine to obtain a pale yellow oily intermediate 2 (NI-FAPI); (3) FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) was dissolved in trifluoroacetic acid, stirred at room temperature, and the solvent was evaporated to dryness to obtain a yellow oily substance. The yellow oily substance was dissolved in anhydrous acetonitrile, anhydrous potassium carbonate was added, stirred at room temperature, and a PEG chain (BocNH(CH 2 CH 2 O) n CH 2 CH 2 Br) was added under ice bath, and the reaction was refluxed overnight. The mixture was filtered by suction, the solvent was evaporated, and purified to obtain a pale yellow oily intermediate 3 (PEG n -FAPI); (4) Dissolve the chelating agent in ultra-dry N,N-dimethylformamide. Under ice bath, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine, stir, add N-Cbz-1,2-diaminoethane. After the reaction is completed, extract with ethyl acetate and saturated brine. Dry the organic phase with anhydrous sodium sulfate and purify by silica gel column chromatography to obtain a yellow solid. Deprotect this yellow solid with hydrogen to obtain a white solid substance; 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxo-6,9-diaza-tetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropionic acid (HBED-CC(tBu) 2 ) is dissolved in ultra-dry N,N-dimethylformamide. Under ice bath, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N’-diisopropylethylamine, stir, add the white solid substance. After the reaction is completed, extract, dry the organic phase, and purify to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu) 2 ); (5) Dissolve the chelating agent in ultradry N,N-dimethylformamide, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under ice bath, stir, and add the intermediate 2 prepared in step (2) or FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester)); after the reaction is completed, extract, dry the organic phase, purify to obtain a pale yellow oil, deprotect with trifluoroacetic acid, and purify by semi-preparative HPLC to obtain Chelator-NI-FAPI or the Chelator-FAPI labeling precursor; (6) Dissolve the chelating agent in ultradry N,N-dimethylformamide. Add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under an ice bath, stir, add the intermediate 1 prepared in step (1), react at room temperature overnight. After the reaction is completed, extract, dry the organic phase, and purify to obtain a yellow oil. Dissolve it in ultradry N,N-dimethylformamide, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under an ice bath, stir, add FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester), react at room temperature overnight. After the reaction is completed, extract, dry the organic phase, and purify to obtain a pale yellow oil. Deprotect with trifluoroacetic acid and purify by semi-preparative HPLC to obtain the NI-Chelator-FAPI labeling precursor; (7) Dissolve the intermediate 4 prepared in step (4) in N,N-dimethylformamide. Under ice bath, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine, stir, add FAPI after removing the Boc protecting group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) or the intermediate 3 prepared in step (3). After the reaction is completed, extract, dry the organic phase, purify to obtain a pale yellow oily substance, remove the protecting group with trifluoroacetic acid, and purify by semi-preparative HPLC to obtain Chelator-HBED-CC-(PEG) n -FAPI labeling precursor; (8) Dissolve the labeling precursors obtained in steps (5), (6), and (7) in dimethyl sulfoxide, add sodium acetate buffer solution and a solution containing a radionuclide, heat to obtain the corresponding radiolabeled product.

4. The method for preparing a radiolabeled FAPI complex according to claim 3, characterized in that: In step (1), the amount of nitroimidazole added is 1 equivalent; the amount of anhydrous potassium carbonate added is 3 - 5 equivalents; the amount of ultradry N,N-dimethylformamide added is 10 - 20 mL; the amount of tert-butyl N-(3-bromopropyl)carbamate added is 1 - 2 equivalents; filter by diatomaceous earth, extract the filtrate with ethyl acetate and saturated brine, dry the organic phase with anhydrous sodium sulfate, and purify by silica gel column chromatography.

5. The method for preparing a radiolabeled FAPI complex according to claim 4, characterized in that: In step (2), the amount of (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 10 - 20 mL; the amounts of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine added are 1 - 2 equivalents; the amount of the yellow oil added is 1 - 1.5 equivalents; the amount of ultra-dry N,N-dimethylformamide added is 5 - 10 mL; the amounts of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine added are 1.3 - 1.5 equivalents; the amount of FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) added is 1 equivalent; it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate and then purified by silica gel column chromatography; it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate and then purified by a rapid purification chromatograph.

6. The method for preparing a radiolabeled FAPI complex according to claim 5, wherein: In step (3), the reflux reaction is carried out at 80 °C; potassium carbonate is removed by suction filtration with diatomaceous earth, the solvent in the filtrate is removed by rotary evaporation under reduced pressure, and the residue is purified by a flash purification chromatograph; the amount of the FAPI (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) added is 1 equivalent; the amount of trifluoroacetic acid (trifluoroacetic acid) added is 3 - 5 mL; the amount of the yellow oily substance added is 1 equivalent; the amount of anhydrous potassium carbonate added is 4 - 5 equivalents; the PEG chain is (2-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamic acid tert-butyl ester, and the amount added is 1.2 - 1.5 equivalents.

7. The method for preparing a radiolabeled FAPI complex according to claim 6, characterized in that: In step (4), the chelating agent is 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-(tert-butoxy)-2-oxoethyl)-1,4-diazacycloheptan-6-yl)pentanoic acid (AAZTA(tBu) 4 ), 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetic acid (DOTA(tBu) 3 ), 5-tert-butoxy-5-oxo-4-(4,7,10-tris(2-tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)pentanoic acid (DOTAGA(tBu) 4 ), 4-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-7-(1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)-1,4,7,10-tetraazacyclododecan-1-yl)-5-tert-butoxy-5-oxopentanoic acid (DOTA(GA) 2 (tBu) 5 ), 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononan-1-yl)acetic acid (NOTA(tBu) 2 ), 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononan-1-yl)-5-tert-butoxy-5-oxopentanoic acid (NODAGA(tBu) 3 ); the addition amount of the chelating agent is 1 equivalent; the addition amount of the ultra-dry N,N-dimethylformamide is 15 - 20 mL; the addition amount of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1 equivalent; the addition amount of N,N'-diisopropylethylamine is 3 equivalents; the addition amount of N-Cbz-1,2-diaminoethane is 1.2 - 1.5 equivalents; extract with ethyl acetate and saturated brine, dry the organic phase with anhydrous sodium sulfate, and purify by silica gel column chromatography to obtain a yellow solid; the addition amount of HBED-CC(tBu) 2 is 1 equivalent; the addition amount of the ultra-dry N,N-dimethylformamide is 10 - 20 mL; the addition amount of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1 equivalent; the addition amount of N,N'-diisopropylethylamine is 3 equivalents; add a white solid substance, after the reaction is completed, extract with ethyl acetate and saturated brine, dry the organic phase with anhydrous sodium sulfate, and purify by a rapid purification chromatograph to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu) 2 ).

8. The method for preparing a radiolabeled FAPI complex according to claim 7, characterized in that: In step (5), the chelating agent is 5-(6-(bis(2-(tert-butoxy)-2-oxoethyl)amino)-1,4-bis(2-(tert-butoxy)-2-oxoethyl)-1,4-diazacycloheptan-6-yl)pentanoic acid (AAZTA(tBu) 4 ), 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetic acid (DOTA(tBu) 3 ), 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropanoic acid (HBED-CC(tBu) 2 ), 5-tert-butoxy-5-oxo-4-(4,7,10-tris(2-tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)pentanoic acid (DOTAGA(tBu) 4 ), 4-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-7-(1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)-1,4,7,10-tetraazacyclododecan-1-yl)-5-tert-butoxy-5-oxopentanoic acid (DOTA(GA) 2 (tBu) 5 ), 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononan-1-yl)acetic acid (NOTA(tBu) 2 ), 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononan-1-yl)-5-tert-butoxy-5-oxopentanoic acid (NODAGA(tBu) 3 ); the addition amount of the chelating agent is 1 - 1.2 equivalents; the addition amount of the super-dry N,N-dimethylformamide is 2 - 5 mL; the addition amount of the 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1 - 1.5 equivalents; the addition amount of the N,N'-diisopropylethylamine is 1.5 - 3 equivalents; the addition amount of the intermediate 2 or the FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) is 1 equivalent; after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate, and purified by a rapid purification chromatograph.

9. The method for preparing a radiolabeled FAPI complex according to claim 8, characterized in that: In step (6), the chelating agent is 3,3'-(((2,2,13,13-tetramethyl-4,11-dioxo-3,12-dioxa-6,9-diazatetradecane-6,9-diyl)bis(methylene))bis(4-hydroxy-3,1-phenylene))dipropanoic acid (HBED-CC(tBu) 2 )、4,4'-(4,10-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,7-diyl)bis(5-tert-butoxy-5-oxopentanoic acid) (DOTA(GA) 2 (tBu) 4 )); The addition amount of the chelating agent is 1 equivalent; the addition amount of the ultra-dry N,N-dimethylformamide is 2 - 10 mL; the addition amount of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1 equivalent; the addition amount of N,N'-diisopropylethylamine is 3 - 5 equivalents; the addition amount of Intermediate 1 is 1 equivalent; after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate, and purified by a rapid purification chromatograph; the addition amounts of the pale yellow oil, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine, and FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester, (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) are all 1 equivalent; the addition amount of the ultra-dry N,N-dimethylformamide is 7 - 10 mL; after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate, and purified by a rapid purification chromatograph; in step (7), the addition amount of Intermediate 4 is 1 equivalent; the addition amount of 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate is 1 - 1.5 equivalents; the addition amount of N,N'-diisopropylethylamine is 4 - 5 equivalents; Intermediate 3 or FAPI after deprotecting the Boc group with trifluoroacetic acid (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)oxy)propyl)-2,5-diazabicyclo[2.2.1] The addition amount of tert-butyl heptane-2-carboxylate and (S)-tert-butyl 4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylate) is 1 - 1.5 equivalents; in step (7), after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried with anhydrous sodium sulfate, and purified by a rapid purification chromatograph; in step (8), the solution containing the radionuclide is [[. 68 Ga] GaCl 3 、 177 Lu] LuCl 3 、 18 F] AlF。 10. Use of the radiolabeled FAPI complex according to any one of claims 1 - 2 in the preparation of a targeted radiodiagnostic and therapeutic drug.

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