Radiolabeled FAPI complex and preparation method and application thereof

Abstract

The invention relates to a radiolabeled FAPI complex as well as a preparation method and application thereof, and belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine diagnosis and treatment. The structural general formula of the FAPI capable of being subjected to radionuclide labeling is as follows: # imgabs0 #, wherein R is a nitroimidazole group or-COOH; x is a chelating group or a chelating structure of a chelating radionuclide; l1 and L2 are linking groups; a is a regulating atom of an FAP targeting group, and A is an H atom or an F atom. The synthesis method of the radionuclide labeling precursor is simple and suitable for labeling of various nuclides, the labeling method is convenient and fast, and the radiochemical purity of the label is high, and the stability is good; the radiolabeled FAPI complex shows high affinity and specificity with FAP in cell experiments, shows high tumor uptake and target-to-non-target ratio in tumor-bearing mice, and is a potential FAP targeted radioactive diagnosis and treatment drug.

Description

Technical Field

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

[0002] Fibroblast activation protein-α (FAP) is not expressed or expressed at low levels in healthy adult tissues, but its expression increases in non-malignant diseases involving tissue remodeling, such as wound healing, cirrhosis, arthritis, atherosclerosis, and fibrosis, and is associated with the degree of disease progression. More importantly, FAP is highly expressed in stromal cancer-associated fibroblasts in more than 90% of epithelial cancers. Therefore, FAP is a potential target for tumor detection and treatment.

[0003] Radionuclide-labeled quinoline-based small molecule inhibitors of 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, the tumor uptake value of FAPI-04 is relatively low, and in particular, the tumor retention time still needs to be improved. Therefore, further increasing the drug uptake and retention time in the tumor while reducing the 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 part of the FAPI structure, but these results have limited improvements on 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 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 method and application are provided. The synthesis method is simple, suitable for multiple radionuclide labeling, and the labeling method is convenient. The radiochemical purity of the label is high and the stability is good. The radionuclide labeled FAPI complex shows high affinity and specificity with FAP in cell experiments, and shows high tumor uptake and target-to-non-target ratio in tumor-bearing mice. The new FAPI that is quickly and efficiently taken up and retained for a long time by the target tissue in the body and quickly cleared by non-target organs will increase the drug uptake dose and retention time at the target site, reduce damage to the blood and non-target organs, and is a potential FAP-targeted radioactive diagnostic and therapeutic drug. Summary of the invention

[0007] One of the purposes of the present invention is to develop a novel radiolabeled FAPI complex, which effectively combines with the FAPI group through different chelating groups and linking groups, thereby increasing the drug uptake in the tumor and prolonging the retention time, thereby overcoming the shortcomings of the radiolabeled FAPI compounds reported in the current literature, namely, low tumor uptake values ​​and short retention times, which are not conducive to the 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 the labeling method is convenient, and the radiochemical purity of the labeled substance is high and the stability is good; the radiolabeled FAPI complex shows high affinity and specificity with FAP in cell experiments, shows very high tumor uptake values ​​and very long tumor retention times in tumor-bearing mice, and exhibits an excellent target-to-non-target ratio, and is a potential novel FAP-targeted radioactive diagnostic and therapeutic drug.

[0008] The above object of the present invention is achieved through the following technical solutions:

[0009] A radiolabeled FAPI complex, the general structural formula of which is shown below:

[0010]

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

[0012]

[0013] X is a chelating group or a chelating structure for chelating radionuclides, selected from any one of the following:

[0014]

[0015] 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+ , 212 Bi 3+ wait;

[0016] L1 and L2 are linking groups, wherein L1 is selected from any one of the following:

[0017]

[0018] In, n is an integer from 0 to 6;

[0019] L2 is selected from any of the following:

[0020]

[0021] A is the regulatory atom of the FAP targeting group, and A is either an H atom or a F atom.

[0022] Preferably, the specific structure is:

[0023]

[0024]

[0025] Another object of the present invention is to provide a method for preparing the above-mentioned radioactive labeled FAPI complex.

[0026] The above object of the present invention is achieved through the following technical solutions:

[0027] A preparation of a radiolabeled FAPI complex, the steps are as follows:

[0028] (1) Nitroimidazole and anhydrous potassium carbonate are dissolved in ultra-dry N,N-dimethylformamide, stirred at room temperature, and tert-butyl N-(3-bromopropyl)carbamate is added. After the reaction is completed, the mixture is filtered and extracted, and the organic phase is dried and purified to obtain a yellow-green oily compound, which is then deprotected with trifluoroacetic acid to obtain a white solid intermediate 1;

[0029] (2) (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid was dissolved in ultra-dry N,N-dimethylformamide, and 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-dihydro ... 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-cyanopyrrolidin-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);

[0030] (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)carbamyl tert-butyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate, (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 dried 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(CH2CH2O) was added under ice bath. n CH2CH2Br), reflux reaction, overnight, suction filtration, distillation of solvent, purification, to obtain light yellow oily intermediate 3 (PEG n -FAPI);

[0031] (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, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, stirred, and a white solid substance was added. After the reaction was completed, extraction was performed, the organic phase was dried, and purified to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu)2);

[0032] (5) dissolving the chelating agent in ultra-dry N,N-dimethylformamide, adding 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under ice bath, stirring, adding intermediate 2 prepared in step (2) or FAPI after deprotection of Boc 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-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, extraction, drying the organic phase, and purification to obtain a light yellow oil, deprotection with trifluoroacetic acid, and purification by semi-preparative HPLC to obtain Chelator-NI-FAPI or Chelator-FAPI labeled precursor;

[0033] (6) 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 1 prepared in step (1) is added, and the reaction is carried out at room temperature overnight. After the reaction is completed, extraction is performed, and the organic phase is 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 are added under ice bath. 7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine, 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, extract after the reaction, dry the organic phase, purify to obtain a light yellow oil, deprotect with trifluoroacetic acid, purify by semi-preparative HPLC to obtain NI-Chelator-FAPI labeled precursor;

[0034] (7) The intermediate 4 prepared in step (4) is dissolved in N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are 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-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) or intermediate 3 prepared in step (3), after the reaction, extraction, drying the organic phase, purification, to obtain a light yellow oily substance, deprotection with trifluoroacetic acid, purification by semi-preparative HPLC, to obtain Chelator-HBED-CC-(PEG) n - FAPI labeled precursors;

[0035] (8) The labeled precursor obtained in steps (5), (6) and (7) is dissolved in dimethyl sulfoxide, a sodium acetate buffer solution and a solution containing a radioactive nuclide are added, and the mixture is heated to obtain a corresponding radioactive labeled product.

[0036] Preferably, in step (1), the nitroimidazole is 2-nitroimidazole, and the amount added is 1 equivalent; the amount of anhydrous potassium carbonate added is 3-5 equivalents; the amount of ultra-dry N,N-dimethylformamide added is 10-20 mL; and the amount of tert-butyl N-(3-bromopropyl)carbamate added is 1-2 equivalents.

[0037] 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.

[0038] Preferably, in step (2), the amount of (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butyloxy)-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.

[0039] 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 amounts of the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine added are 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 (S)-4-(3-((4-((2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylic acid tert-butyl ester) added is 1 equivalent.

[0040] 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.

[0041] 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 a rapid purification chromatograph.

[0042] 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-cyanopyrrolidinyl)- 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.

[0043] 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.

[0044] 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-tetraazacyclododecane-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).

[0045] 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.

[0046] 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.

[0047] 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)

[0048] 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-butyloxy-5-oxo-4-(4,7,10-tri( 2-tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-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-tetraazacyclododecane-1-yl)-5-tert-butoxy-5-oxopentanic 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).

[0049] 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-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 added in an amount of 1 equivalent; 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 using a speed purification chromatograph.

[0050] 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).

[0051] Preferably, in step (6), the chelating agent is added in an amount of 1 equivalent; the ultra-dry N,N-dimethylformamide is added in an amount of 2-10 mL; the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate is added in an amount of 1 equivalent; the N,N'-diisopropylethylamine is added in an amount of 3-5 equivalents; and the intermediate 1 is added in an amount of 1 equivalent.

[0052] Preferably, in step (6), after the reaction is completed, extraction is performed with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a rapid purification chromatograph.

[0053] Preferably, in step (6), the light yellow oil, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine and FAPI after deprotection from Boc 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-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide The amount of tert-butyl 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-carboxylate, and (S)-4-(3-((4-((2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)carbamoyl)quinolin-6-yl)(methyl)amino)propyl)piperazine-1-carboxylate) added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 7-10 mL.

[0054] Preferably, in step (6), after the reaction is completed, extraction is performed with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a rapid purification chromatograph.

[0055] 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-piperazinyl)propoxy]-N -[2-(2-cyanopyrrolidin-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 added in an amount of 1-1.5 equivalents.

[0056] Preferably, in step (7), after the reaction is completed, extraction is performed with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified using a rapid purification chromatograph.

[0057] Preferably, in step (8), the solution containing radionuclides is [ 68 Ga]GaCl3, [ 177 Lu]LuCl3, [ 18 F]AlF.

[0058] Another object of the present invention is to provide the application of the above-mentioned radioactive labeled FAPI complex.

[0059] The above object of the present invention is achieved through the following technical solutions:

[0060] Application of radiolabeled FAPI complexes in FAP-targeted radiotherapy.

[0061] Beneficial effects:

[0062] The novel radiolabeled FAPI complex of the present invention effectively combines with the FAPI group through different chelating groups and linking groups, thereby improving the uptake and retention time of the drug in the tumor. The novel radiolabeled FAPI complex has a simple synthesis method; it is suitable for multiple nuclide labeling, and the labeling method is convenient, and the labeled substance has high radiochemical purity and good stability; the radionuclide labeled compound shows high affinity and specificity with FAP in cell experiments, and shows high tumor uptake and target-to-non-target ratio in tumor-bearing mice, and is a potential FAP-targeted radioactive diagnostic and therapeutic drug.

[0063] The present invention is further described below through the accompanying drawings and specific implementation modes, but it is not intended to limit the protection scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0064] Figure 1 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-AAZTA-FAPI-04.

[0065] Figure 2 [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-AAZTA-FAPI-04.

[0066] Figure 3 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-AAZTA-NI-FAPI-04.

[0067] Figure 4 [ 177

[0045] Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-AAZTA-NI-FAPI-04.

[0068] Figure 5 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-DOTA-NI-FAPI-04.

[0069] Figure 6 [ 177

[0045] Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-NI-FAPI-04.

[0070] Figure 7 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-NI-FAPI-04.

[0071] Figure 8 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-NI-HBED-CC-FAPI-04.

[0072] Fig. 9 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-NI-HBED-CC-FAPI-02.

[0073] Fig.10 DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-FAPI-02.

[0074] Fig.11 [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-02.

[0075] Fig.12 DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-FAPI-04.

[0076] Fig.13 [ 177Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-FAPI-04.

[0077] Fig.14 DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-HBED-CC-PEG3-FAPI-02.

[0078] Fig.15 [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02.

[0079] Fig.16 DOTA-[ 68 Radioactive HPLC spectrum of the labeling reaction solution of [Ga]Ga-HBED-CC-PEG3-FAPI-04.

[0080] Fig.17 [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04.

[0081] Fig.18 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 in HT1080-FAP cells in vitro.

[0082] Fig.19 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-[ 68Ga]Ga-HBED-CC-PEG3-FAPI-04(D) and [ 68 (E) Uptake and blocking results of Ga]Ga-DOTA-FAPI-04 in HT1080-FAP cells in vitro for 60 min.

[0083] Fig. 20 For the application example 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.

[0084] Fig.21 For the application example 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 (E) Uptake and blocking results of Lu]Lu-DOTA-FAPI-04 in HT1080-FAP cells in vitro for 60 min.

[0085] Fig. 22 For the present invention, in Example 3 [ 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.

[0086] Fig.23 For the present invention, in Example 3 [ 177 Lu]Lu-AAZTA-FAPI-04(F), [ 177Lu]Lu-AAZTA-NI-FAPI-04(G), [ 177 Lu]Lu-DOTA-NI-FAPI-04(H) and [ 177 (E) Uptake and blocking results of Lu]Lu-DOTA-FAPI-04 in HT1080-FAP cells in vitro for 60 min.

[0087] Fig.24 For the present invention, in Example 4, [ 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 in HT1080-FAP cells in vitro.

[0088] Fig.25 For the present invention, in Example 4, [ 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.

[0089] Fig.26 For the application example 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 [ 68In vivo PET / CT imaging results of U87MG tumor-bearing mice with Ga]Ga-DOTA-FAPI-04 (T: tumor; G: Gallbladder). DETAILED DESCRIPTION

[0090] Unless otherwise specified, the reagents and raw materials used in the preparation methods and detection methods described in the following examples and comparative examples are all commercially available products, and the equipment used is all commonly used equipment; the concentrations and ratios described are all weight and amount of substance units.

[0091] Example 1: 68 Preparation of Ga]Ga-AAZTA-FAPI-04

[0092] Step 1: Synthesis of AAZTA-FAPI-04

[0093] The synthetic route is as follows:

[0094]

[0095] The specific steps include:

[0096] (1) Synthesis of Compound 1

[0097] 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 filtrate was evaporated under reduced pressure to remove the solvent and purified by silica gel column chromatography (dichloromethane / methanol / ammonia water, v / v / v=20 / 1 / 0.1) to obtain an orange-yellow compound 1 (64.70 mg, 0.06 mmol) with a yield of 67%;

[0098] Structure confirmation of compound 1:

[0099] HRMS C 58 H 88 N9O 12 F2[M+H] + Theoretical molecular weight is 1140.6526, and the measured molecular weight is 1140.6520.

[0100] 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).

[0101] (2) Synthesis of compound AAZTA-FAPI-04

[0102] 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 removed by vacuum rotary evaporation. 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: 0-12 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 by a lyophilizer to obtain a white solid compound AAZTA-FAPI-04, which was determined by LC-MS to have a purity of >97%.

[0103] Structural confirmation of AAZTA-FAPI-04:

[0104] HRMS C 42 H 56 N9O 12 F2[M+H] + Theoretical molecular weight is 916.4011, and the measured molecular weight is 916.4016.

[0105] Step 2: Ga-68 labeling of AAZTA-FAPI-04:

[0106] The Ga-68 marked route is as follows:

[0107]

[0108] The marking method is as follows:

[0109] 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 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the mixed solution of radioligand sodium acetate. After mixing evenly, react at 50°C for 10 minutes. Determine its radiochemical purity by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-AAZTA-FAPI-04.

[0110] like Figure 1 As shown, it is [ 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%.

[0111] Example 2: 177 Preparation of Lu]Lu-AAZTA-FAPI-04

[0112] Step 1: Synthesis of AAZTA-FAPI-04:

[0113] The synthetic route and method are the same as step 1 in Example 1;

[0114] Step 2: Lu-177 labeling of AAZTA-FAPI-04:

[0115] The Lu-177 marked route is as follows:

[0116]

[0117] The marking method is as follows:

[0118] 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;

[0119] like Figure 2 As shown, it is [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-AAZTA-FAPI-04 shows that [ 177 The radiochemical purity of Lu]Lu-AAZTA-FAPI-04 was greater than 95%.

[0120] Example 3: 68 Preparation of Ga]Ga-AAZTA-NI-FAPI-04

[0121] Step 1: Synthesis of AAZTA-NI-FAPI-04:

[0122] The synthetic route is as follows:

[0123]

[0124] The specific steps include:

[0125] (1) Synthesis of Compound 2

[0126] 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 over a 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 anhydrous sodium sulfate solid was removed. The filtrate was rotary evaporated 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 obtain a yellow-green oil 2 (2.36 g, 8.74 mmol), with a yield of 87%;

[0127] The structure of compound 2 was confirmed:

[0128] HRMS C 11 H 19 N4O4[M+H] + Theoretical molecular weight 271.1400, measured molecular weight 271.1408;

[0129] 1H 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);

[0130] (2) Synthesis of Compound 3

[0131] 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-fluorene-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, and the mixture was stirred at room temperature overnight, and then 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, and purified by silica gel column chromatography (dichloromethane / methanol / ammonia water, v / v / v=25 / 1 / 0.1) to obtain a light yellow solid compound 3 (1.5 g, 2.60 mmol), with a yield of 74%;

[0132] The structure of compound 3 was confirmed:

[0133] HRMS C 30 H 36 N5O7[M+H] + Theoretical molecular weight 578.2609, measured molecular weight 578.2609;

[0134] 1H 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);

[0135] (2) Synthesis of Compound 4

[0136] 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. 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 mixture 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 dimethylformamide was added to the above reaction solution, reacted 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, anhydrous sodium sulfate was removed, the filtrate was evaporated under reduced pressure, the solvent was removed, and the residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=15 / 1 / 0.1) to obtain a light yellow oily compound 4 (205.6 mg, 0.21 mmol), yield: 81%;

[0137] The structure of compound 4 was confirmed:

[0138] HRMS C 50 H 54 N 11O9F2[M+H] + Theoretical molecular weight 990.4068, measured molecular weight 990.4076;

[0139] 1 H NMR (600MHz, CD3OD) δ8.69(d,J=4.4Hz,1H),7.90(dd,J=16.1,5.8Hz,2H),7.73(d,J=7.5Hz,2H),7.60(dd,J=11.1 ,7.6Hz,2H),7.51(d,J=4.4Hz,1H),7.45(s,1H),7.40(dd,J=9.2,2.6Hz,1H),7.32(t,J=7.4Hz,2H),7.25(ddd,J=

[0140] 10.1,5.5,2.1Hz,2H),7.03(s,1H),5.07(dd,J=9.3,2.7Hz,1H),4.57(dd,J=9.7,3.9Hz,1H),4.44-4.37(m,2H),4.34(dd,J= 10.5,6.9Hz,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= 3.1,1.5Hz,3H),3.15-3.07(m,1H),2.85(ddt,J=14.0,9.5,6.9Hz,1H),2.73(t,J=13.6Hz,1H),2.61-2.44(m,5H),2.32-2.20 (m,2H),1.98(dtd,J=20.6,13.8,6.9Hz,5H),1.74(td,J=15.4,6.0Hz,1H),1.33(dd,J=26.1,13.4Hz,1H),1.29-1.21(m,3H);

[0141] (2) Synthesis of Compound 5

[0142] 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, 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, 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, the filtrate was evaporated under reduced pressure, the solvent was removed, and the mixture was purified by rapid purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain yellow oily compound 5 (29 mg, 0.02 mmol), yield: 40%;

[0143] HRMS C 69 H 103 N 14 O 16 F2[M+H] + Theoretical molecular weight 1421.7639, measured molecular weight 1421.7646;

[0144] 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);

[0145] (2) Synthesis of compound AAZTA-NI-FAPI-04

[0146] 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 by a lyophilizer to obtain AAZTA-NI-FAPI-04 as a white solid.

[0147] Structure confirmation of compound AAZTA-NI-FAPI-04:

[0148] HRMS C 53 H 70 N 14 O 16 F2[M+H] + Theoretical molecular weight 1197.5135, measured molecular weight 1197.5132;

[0149] Step 2: Ga-68 labeling of AAZTA-NI-FAPI-04:

[0150] The Ga-68 marked route is as follows:

[0151]

[0152] The marking method is as follows:

[0153] 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 Take 300 μL of Ga]GaCl3 hydrochloric acid solution and add it to the mixed solution of radioligand sodium acetate. After mixing evenly, react at 50°C for 10 minutes. Determine its radiochemical purity by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-AAZTA-NI-FAPI-04;

[0154] like Figure 3 As shown, it is [ 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%;

[0155] Example 4: 177 Preparation of Lu]Lu-AAZTA-NI-FAPI-04

[0156] Step 1: Synthesis of AAZTA-NI-FAPI-04:

[0157] The synthetic route and method are the same as step 1 in Example 3;

[0158] Step 2: Lu-177 labeling of AAZTA-NI-FAPI-04:

[0159] The Lu-177 marked route is as follows:

[0160]

[0161] The marking method is as follows:

[0162] 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 [ 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-NI-FAPI-04;

[0163] like Figure 4 As shown, it is [ 177 The radioactive HPLC spectrum of the labeling 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%.

[0164] Example 5: 68 Preparation of Ga]Ga-DOTA-NI-FAPI-04

[0165] Step 1: Synthesis of DOTA-NI-FAPI-04:

[0166] The synthetic route is as follows:

[0167]

[0168] The specific steps include:

[0169] (1) Synthesis of Compound 6

[0170] 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 monitored 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, 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 at room temperature for 2 hours. Stir for 30 minutes to obtain a reaction solution, add a light yellow oily intermediate (38 mg, 0.05 mmol) dissolved in 2 mL of ultra-dry N,N-dimethylformamide to the above reaction solution, react at room temperature overnight, wash with ethyl acetate (10 mL×3) and saturated brine (10 mL×2), collect the organic phase and dry it over anhydrous sodium sulfate, filter and remove anhydrous sodium sulfate, evaporate the filtrate under reduced pressure, remove the solvent, and purify it with a flash purification chromatograph (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to obtain a yellow oily compound 6 (21.5 mg, 0.02 mmol), yield: 40%;

[0171] The structure of compound 6 was confirmed:

[0172] HRMS C 63 H 93 N 15 O 14 F2Na[M+Na] + Theoretical molecular weight 1344.6886, measured molecular weight 1344.6878;

[0173] 1H 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);

[0174] (2) Synthesis of compound DOTA-NI-FAPI-04

[0175] Compound 6 (21.5 mg, 0.02 mmol) was dissolved in 2 mL of trifluoroacetic acid, stirred at room temperature for 2 hours, and evaporated under reduced pressure to remove the solvent. 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 by a lyophilizer to obtain a white solid compound DOTA-NI-FAPI-04.

[0176] HRMS C 51 H 69 N 15 O 14 F2[M+H] + Theoretical molecular weight 1154.5189, measured molecular weight 1154.5188;

[0177] Step 2: Ga-68 labeling of DOTA-NI-FAPI-04:

[0178] 68 Ga marked routes are as follows:

[0179]

[0180] The marking method is as follows:

[0181] 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 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 95℃ for 15 minutes, cool to room temperature, and measure its radiochemical purity by high performance liquid chromatography with a radioactivity detector. The radiochemical purity of [ 68 Ga]Ga-DOTA-NI-FAPI-04;

[0182] like Figure 5 As shown, it is [ 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%;

[0183] Example 6: 177 Preparation of Lu]Lu-DOTA-NI-FAPI-04

[0184] Step 1: Synthesis of DOTA-NI-FAPI-04:

[0185] The synthetic route and synthetic method are the same as step 1 in Example 5;

[0186] Step 2: Lu-177 marking of DOTA-NI-FAPI-04:

[0187] The Lu-177 marked route is as follows:

[0188]

[0189] The marking method is as follows:

[0190] 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;

[0191] like Figure 6 As shown, it is [ 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%.

[0192] Example 7: 68 Preparation of Ga]Ga-HBED-CC-NI-FAPI-04

[0193] Step 1: Synthesis of HBED-CC-NI-FAPI-04:

[0194] The synthetic route is as follows:

[0195]

[0196] The specific steps include:

[0197] (1) Synthesis of Compound 7

[0198] 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 hexafluorophosphoric acid was added under ice bath. Acid ester (27 mg, 0.07 mmol) and N, N'-diisopropylethylamine (13 mg, 0.1 mmol) were added to the reaction solution, stirred in an ice bath for 30 minutes, and a light yellow intermediate (38 mg, 0.05 mmol) dissolved in 2 mL of ultra-dry N, N-dimethylformamide was added to the reaction solution, and the reaction was allowed to proceed 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, and anhydrous sodium sulfate was removed. The filtrate was evaporated under reduced pressure to remove the solvent, and purified by a rapid purification chromatograph (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain a yellow oily compound 7 (21 mg, 0.02 mmol) with a yield of 40%.

[0199] The structure of compound 7 was confirmed:

[0200] HRMS C 69 H 90 N 13 O 16 F2[M+H] + Theoretical molecular weight 1394.6591, measured molecular weight 1394.6599;

[0201] 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);

[0202] (2) Synthesis of compound HBED-CC-NI-FAPI-04

[0203] Compound 7 (21 mg, 0.02 mmol) was dissolved in 2 mL of trifluoroacetic acid, stirred at room temperature for 2 hours, and evaporated under reduced pressure to remove the solvent. 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 by a lyophilizer to obtain a white solid compound HBED-CC-NI-FAPI-04.

[0204] The structure of compound HBED-CC-NI-FAPI-04 was confirmed:

[0205] HRMS C 61 H 73 F2N 13 O 16 [M+H] + Theoretical molecular weight 1282.5339, measured molecular weight 1282.5331;

[0206] Step 2: Ga-68 labeling of HBED-CC-NI-FAPI-04:

[0207] The Ga-68 marked route is as follows:

[0208]

[0209] The marking method is as follows:

[0210] 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 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 50℃ for 10 minutes, cool to room temperature, and measure its radiochemical purity by high performance liquid chromatography with a radioactive detector. The radiochemical purity of [ 68 Ga]Ga-HBED-CC-NI-FAPI-04;

[0211] like Figure 7 As shown, it is [ 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%.

[0212] Example 8: 68 Preparation of Ga]Ga-NI-HBED-CC-FAPI-04

[0213] Step 1: Synthesis of NI-HBED-CC-FAPI-04:

[0214] The synthetic route is as follows:

[0215]

[0216] The specific steps include:

[0217] (1) Synthesis of Compound 8

[0218] 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, 0.48 mmol) was added under ice bath. , 0.48mmol) and N,N'-diisopropylethylamine (187mg, 1.45mmol), stirred in an ice bath for 30 minutes to obtain a reaction solution, a white solid intermediate dissolved in 2mL ultra-dry N,N-dimethylformamide was added to the above reaction solution, reacted at room temperature for 5 hours, the obtained reaction solution was washed with ethyl acetate (20mL×3) and saturated brine (20mL×2), the organic phase was collected and dried over anhydrous sodium sulfate, filtered to remove anhydrous sodium sulfate, the filtrate was evaporated under reduced pressure, the solvent was removed, and it was purified by rapid purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to obtain a light yellow oily compound 8 (55mg, 0.07mmol); yield: 15%;

[0219] The structure of compound 8 was confirmed:

[0220] HRMS C 40 H 56 N6O 11 [M+H] + Theoretical molecular weight 797.4085, measured molecular weight 797.4081;

[0221] 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);

[0222] (2) Synthesis of Compound 9

[0223] 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, reacted at room temperature for 5 hours, 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, anhydrous sodium sulfate was removed, the filtrate was evaporated under reduced pressure, the solvent was removed, and purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v = 10 / 1 / 0.1) to obtain a light yellow oily compound 9 (31.7 mg, 0.025 mmol); yield: 36%;

[0224] HRMS C 64 H 82 N 12 O 143 F2[M+H] +Theoretical molecular weight 1265.617, measured molecular weight 1265.6169;

[0225] 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);

[0226] (3) Synthesis of compound NI-HBED-CC-FAPI-04

[0227] Compound 9 (31.7 mg, 0.025 mmol) was dissolved in 2 mL of trifluoroacetic acid, stirred at room temperature for 30 minutes, and evaporated under reduced pressure to remove the solvent. 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 by a lyophilizer to obtain NI-HBED-CC-FAPI-04 as a white solid.

[0228] The structure of compound NI-HBED-CC-FAPI-04 was confirmed:

[0229] HRMS C 56 H 67 N 12 O 13 F2[M+H] +Theoretical molecular weight 1153.4913, measured molecular weight 1153.4912;

[0230] Step 2: Ga-68 labeling of NI-HBED-CC-FAPI-04:

[0231] The Ga-68 marked route is as follows:

[0232]

[0233] The marking method is as follows:

[0234] 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 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 50℃ for 10 minutes, cool to room temperature, and measure its radiochemical purity by high performance liquid chromatography with a radioactive detector. The radiochemical purity of [ 68 Ga]Ga-NI-HBED-CC-FAPI-04;

[0235] like Figure 8 As shown, it is [ 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%.

[0236] Example 9: 68 Preparation of Ga]Ga-NI-HBED-CC-FAPI-02

[0237] Step 1: Synthesis of NI-HBED-CC-FAPI-02:

[0238] The synthetic route is as follows:

[0239]

[0240] The specific steps include:

[0241] (1) Synthesis of Compound 10

[0242] (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, stirred at room temperature for 30 minutes, and evaporated under reduced pressure to remove the solvent 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. ethylamine (33 mg, 0.26 mmol), 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, 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, anhydrous sodium sulfate was removed, the filtrate was evaporated under reduced pressure, the solvent was removed, and it was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain a light yellow oily compound 10 (38.4 mg, 0.031 mmol), yield: 48%;

[0243] The structure of compound 10 was confirmed:

[0244] HRMS C 64 H 85 N 12 O 13 [M+H] + Theoretical molecular weight 1229.6353, measured molecular weight 1229.6356;

[0245] (2) Synthesis of compound NI-HBED-CC-FAPI-02

[0246] Compound 10 (38.4 mg, 0.031 mmol) was dissolved in 2 mL of trifluoroacetic acid, stirred at room temperature for 1 hour, and evaporated under reduced pressure to remove the solvent. 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: 0-25 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 by a lyophilizer to obtain NI-HBED-CC-FAPI-02 as a white solid.

[0247] The structure of compound NI-HBED-CC-FAPI-02 was confirmed:

[0248] HRMS C 56 H 68 N 12 O 13 [M+H] + Theoretical molecular weight 1117.5101, measured molecular weight 1117.5104;

[0249] Step 2: Ga-68 labeling of NI-HBED-CC-FAPI-02:

[0250] The Ga-68 marked route is as follows:

[0251]

[0252] The marking method is as follows:

[0253] 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 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 50℃ for 10 minutes, cool to room temperature, and measure its radiochemical purity by high performance liquid chromatography with a radioactive detector. The radiochemical purity of [ 68 Ga]Ga-NI-HBED-CC-FAPI-02;

[0254] like Fig. 9 As shown, it is [ 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%.

[0255] Example 10: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-FAPI-02

[0256] Step 1: Synthesis of DOTA-HBED-CC-FAPI-02:

[0257] The synthetic route is as follows:

[0258]

[0259] The specific steps include:

[0260] (1) Synthesis of Compound 11

[0261] 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, and 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, and the mixture was stirred for 30 minutes while keeping ice bath. The mixture was added with N-Cbz-1,2-diaminoethane (408 mg, 2.1 mmol) dissolved in 5 mL of ultra-dry N,N-dimethylformamide, and the mixture was reacted at room temperature for 5 hours. 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, and purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=20 / 1 / 0.1) to obtain yellow solid compound 11 (1.02 g, 1.36 mmol), with a yield of 78%;

[0262] The structure of compound 11 was confirmed:

[0263] HRMS C 38 H 65 N6O9[M+H] + Theoretical molecular weight 749.4807, measured molecular weight 749.4802;

[0264] 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);

[0265] (2) Synthesis of Compound 12

[0266] 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 in a hydrogen atmosphere overnight. The resulting reaction solution was filtered with diatomaceous earth, and the filtrate was evaporated under reduced pressure to remove the solvent to obtain white solid compound 12 (852 mg, 1.38 mmol), yield: >99%; the next step of the reaction was carried out without purification;

[0267] (3) Synthesis of Compound 13

[0268] 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, N,N'-diisopropylethylamine (300 mg, 2.3 mmol) was added under ice bath conditions, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2 89 mg, 0.76 mmol), 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, reacted at room temperature for 5 hours, and most of the N, N-dimethylformamide was removed by pressure rotary evaporation. 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, and 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 colorless solid compound 13 (234 mg, 0.19 mmol), yield: 24%;

[0269] The structure of compound 13 was confirmed:

[0270] HRMS C 64 H 105 N8O 16 [M+H] + Theoretical molecular weight 1241.7643, measured molecular weight 1241.7652;

[0271] 1H 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);

[0272] (4) Synthesis of Compound 14

[0273] 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 spin-dried to obtain 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 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 purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain a light yellow oily compound 14 (15 mg, 0.009 mmol) with a yield of 31%.

[0274] The structure of compound 14 was confirmed:

[0275] HRMS C 88 H 133 N 14 O 18 [M+H] + Theoretical molecular weight 1673.9916, measured molecular weight 1673.9922;

[0276] (5) Synthesis of compound DOTA-HBED-CC-FAPI-02

[0277] In a 25 mL round-bottom flask, compound 14 (15 mg, 0.009 mmol) was dissolved in 4 mL of trifluoroacetic acid, stirred at room temperature for 2.5 hours, and evaporated under reduced pressure to remove the solvent. 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: 0-15 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 by a lyophilizer to obtain a white solid compound DOTA-HBED-CC-FAPI-02, and the purity was >95% as determined by LC-MS.

[0278] The structure of the compound DOTA-HBED-CC-FAPI-02 was confirmed:

[0279] HRMS C 68 H 93 N 14 O 18 [M+H] + Theoretical molecular weight 1393.6786, measured molecular weight 1393.6784;

[0280] Step 2: Ga-68 labeling of DOTA-HBED-CC-FAPI-02:

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

[0282]

[0283] The marking method is as follows:

[0284] 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;

[0285] like Fig.10 As shown, it is DOTA-[ 68 The radioactive HPLC spectrum of the labeling 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%.

[0286] Example 11: 177 Preparation of Lu]Lu-DOTA-HBED-CC-FAPI-02

[0287] Step 1: Synthesis of DOTA-HBED-CC-FAPI-02:

[0288] The synthetic route and synthetic method are the same as step 1 in Example 10;

[0289] Step 2: Lu-177 labeling of DOTA-HBED-CC-FAPI-02:

[0290] The Lu-177 marked route is as follows:

[0291]

[0292] The marking method is as follows:

[0293] 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;

[0294] like Fig.11 As shown, it is [ 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%.

[0295] Example 12: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-FAPI-04

[0296] Step 1: Synthesis of DOTA-HBED-CC-FAPI-04:

[0297] The synthetic route is as follows:

[0298]

[0299] (1) Synthesis of Compound 15

[0300] 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 spin-dried to obtain 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, reacted at room temperature for 5 hours, and 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 reduced pressure rotary evaporation, and the residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=8 / 1 / 0.1) to obtain a light yellow oily compound 15 (29 mg, 0.017 mmol), yield: 68%;

[0301] The structure of compound 15 was confirmed:

[0302] HRMS C 88 H 132 N 14 O 18 F2[M+2H] 2+ Theoretical molecular weight 855.49, measured molecular weight 855.4894;

[0303] (2) Synthesis of compound DOTA-HBED-CC-FAPI-04

[0304] In a 25 mL round-bottom flask, compound 15 (29 mg, 0.017 mmol) was dissolved in 4 mL trifluoroacetic acid, stirred at room temperature for 2.5 hours, and evaporated under reduced pressure to remove the solvent. 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: 0-15 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 by a lyophilizer to obtain a white solid compound DOTA-HBED-CC-FAPI-04, and the purity was determined by LC-MS to be >95%;

[0305] The structure of compound DOTA-HBED-CC-FAPI-04 was confirmed:

[0306] HRMS C 68 H 91 N 14 O 18 F2[M+H] + Theoretical molecular weight 1429.6598, measured molecular weight 1429.6592;

[0307] Step 2: Ga-68 labeling of DOTA-HBED-CC-FAPI-04:

[0308] The Ga-68 marked route is as follows:

[0309]

[0310] The marking method is as follows:

[0311] 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;

[0312] like Fig.12 As shown, it is DOTA-[ 68The radioactive HPLC spectrum of the labeling reaction solution of DOTA-[Ga]Ga-HBED-CC-FAPI-04 shows that 68 The radiochemical purity of Ga]Ga-HBED-CC-FAPI-04 was greater than 95%.

[0313] Example 13: 177 Preparation of Lu]Lu-DOTA-HBED-CC-FAPI-04

[0314] Step 1: Synthesis of DOTA-HBED-CC-FAPI-04:

[0315] The synthetic route and synthetic method are the same as step 1 in Example 12;

[0316] Step 2: Lu-177 labeling of DOTA-HBED-CC-FAPI-04:

[0317] The Lu-177 marked route is as follows:

[0318]

[0319] The marking method is as follows:

[0320] 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;

[0321] like Fig.13 As shown, it is [ 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%.

[0322] Example 14: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-PEG3-FAPI-02

[0323] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-02:

[0324] The synthetic route is as follows:

[0325]

[0326] The specific steps include:

[0327] (1) Synthesis of Compound 16

[0328] 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, anhydrous potassium carbonate (216 mg, 1.56 mmol) was added, and 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 diatomaceous earth filtration. The filtrate was evaporated under reduced pressure to remove the solvent, and the residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to obtain a light yellow oily compound 16 (191 mg, 0.26 mmol) with a yield of 67%.

[0329] The structure of compound 16 was confirmed:

[0330] HRMS C 37 H 56 N7O8[M+H] + Theoretical molecular weight 726.419, measured molecular weight 726.4179;

[0331] 1H 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);

[0332] (2) Synthesis of Compound 17

[0333] In a 50 mL round-bottom flask, compound 16 (32 mg, 0.044 mmol) was dissolved in 3 mL 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 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 ultra-dry N,N-dimethylformamide. Formamide was added to the above reaction solution, and the reaction was carried out at room temperature for 5 hours. 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 purification chromatography (dichloromethane / methanol / ammonia water, v / v / v = 8 / 1 / 0.1) to obtain a light yellow oily compound 17 (31 mg, 0.017 mmol), with a yield of 57%;

[0334] The structure of compound 17 was confirmed:

[0335] HRMS C 96 H 150 N 15 O 21 Na[M+H+Na] 2+ Theoretical molecular weight 936.0508, measured molecular weight 936.0511;

[0336] (3) Synthesis of compound DOTA-HBED-CC-PEG3-FAPI-02

[0337] In a 25 mL round-bottom flask, compound 17 (31 mg, 0.017 mmol) was dissolved in 4 mL of trifluoroacetic acid, stirred at room temperature for 2.5 hours, and evaporated under reduced pressure to remove the solvent. 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: 0-15 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 by a lyophilizer to obtain a white solid compound DOTA-HBED-CC-PEG3-FAPI-02, and the purity was determined by LC-MS to be >95%;

[0338] The structure of the compound DOTA-HBED-CC-PEG3-FAPI-02 was confirmed:

[0339] HRMS C 76 H 110 N 15 O 21 [M+H] + Theoretical molecular weight 1568.7995, measured molecular weight 1568.7989;

[0340] Step 2: Ga-68 labeling of DOTA-HBED-CC-PEG3-FAPI-02:

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

[0342]

[0343] The marking method is as follows:

[0344] 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.

[0345] like Fig.14 As shown, it is DOTA-[ 68 The radioactive HPLC spectrum of the labeled reaction solution of DOTA-[Ga]Ga-HBED-CC-PEG3-FAPI-02 shows that 68 The radiochemical purity of Ga]Ga-HBED-CC-PEG3-FAPI-02 was greater than 95%.

[0346] Example 15: 177 Preparation of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02

[0347] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-02:

[0348] The synthetic route and synthetic method are the same as step 1 in Example 14;

[0349] Step 2: Lu-177 labeling of DOTA-HBED-CC-PEG3-FAPI-02:

[0350] The Lu-177 marked route is as follows:

[0351]

[0352] The marking method is as follows:

[0353] The DOTA-HBED-CC-PEG3-FAPI-02 obtained in step 1 was prepared into a 1 μg / μL precursor solution with dimethyl sulfoxide. 31 μL of the precursor solution was placed in a 10 mL vial, 12 μL of 3 M sodium acetate solution was added, 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.

[0354] like Fig.15 As shown, it is [ 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%.

[0355] Example 16: DOTA-[ 68 Preparation of Ga]Ga-HBED-CC-PEG3-FAPI-04

[0356] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-04:

[0357] The synthetic route is as follows:

[0358]

[0359] The specific steps include:

[0360] (1) Synthesis of Compound 18

[0361] 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 ol), stirred at room temperature for 30 minutes, added tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamate (82 mg, 0.23 mmol), the mixed solution was refluxed at 80°C overnight, potassium carbonate was removed by diatomaceous earth filtration, the filtrate was evaporated under reduced pressure to remove the solvent, and the residue was purified by flash purification chromatography (dichloromethane / methanol / ammonia water, v / v / v=10 / 1 / 0.1) to obtain light yellow oily compound 18 (131 mg, 0.17 mmol), yield: 94%;

[0362] The structure of compound 18 was confirmed:

[0363] HRMS C 37 H 54 N7O8F2[M+H] + Theoretical molecular weight 762.3996, measured molecular weight 762.3992;

[0364] 1H 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);

[0365] (2) Synthesis of Compound 19

[0366] 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 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 a light yellow oily compound 19 (28 mg, 0.015 mmol). The yield was 58%.

[0367] The structure of compound 19 was confirmed:

[0368] HRMS C 96 H 149 N 15 O 21 F2[M+2H] 2+ Theoretical molecular weight 943.0504, measured molecular weight 943.0520;

[0369] (3) Synthesis of compound DOTA-HBED-CC-PEG3-FAPI-04

[0370] In a 25 mL round-bottom flask, compound 19 (28 mg, 0.015 mmol) was dissolved in 4 mL of trifluoroacetic acid, stirred at room temperature for 2.5 hours, and evaporated under reduced pressure to remove the solvent. 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: 0-15 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 by a lyophilizer to obtain a white solid compound DOTA-HBED-CC-PEG3-FAPI-04, and the purity was determined by LC-MS to be >95%;

[0371] The structure of the compound DOTA-HBED-CC-PEG3-FAPI-04 was confirmed:

[0372] HRMS C 76 H 108 N 15 O 21 F2[M+H] + Theoretical molecular weight 1604.7806, measured molecular weight 1604.7778;

[0373] Step 2: Ga-68 labeling of DOTA-HBED-CC-PEG3-FAPI-04:

[0374] The Ga-68 marked route is as follows:

[0375]

[0376] The marking method is as follows:

[0377] 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;

[0378] like Fig.16As shown, it is DOTA-[ 68 The radioactive HPLC spectrum of the labeling reaction solution of DOTA-[ 68 The radiochemical purity of Ga]Ga-HBED-CC-PEG3-FAPI-04 was greater than 95%.

[0379] Example 17: 177 Preparation of Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04

[0380] Step 1: Synthesis of DOTA-HBED-CC-PEG3-FAPI-04:

[0381] The synthetic route and synthetic method are the same as step 1 in Example 16;

[0382] Step 2: Lu-177 labeling of DOTA-HBED-CC-PEG3-FAPI-04:

[0383] The Lu-177 marked route is as follows:

[0384]

[0385] The marking method is as follows:

[0386] 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.

[0387] like Fig.17 As shown, it is [ 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%.

[0388] Application Example 1

[0389] 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:

[0390] HT1080-FAP cells were prepared at 1.25 × 10 6 400 μL of the 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-[ 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 solution and incubated in a 37°C incubator. At 10, 30, 60, 90 and 120 minutes of incubation in each group, PBS solution was used to terminate its uptake, cells were lysed with 1M NaOH, and the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count; for blocking experiments, excess unlabeled DOTA-FAPI-04 was used to incubate with the corresponding radiolabeled drugs. After 60 minutes of incubation, PBS solution was used to terminate its uptake, cells were lysed with 1M NaOH, and the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0391] like Fig.18 As shown, 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 [ 68Ga]Ga-DOTA-FAPI-04 uptake-time curve (n=3); Fig.19 As shown, in the application example 1 of the present invention, the in vitro HT1080-FAP cells are sensitive to DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-02, DOTA-[ 68 Ga]Ga-HBED-CC-FAPI-04,

[0392] 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).

[0393] The results of in vitro cell uptake experiments showed that 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 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 Ga]Ga-DOTA-FAPI-04, the uptake of all drugs could be blocked by excess DOTA-FAPI-04, indicating that all drugs specifically targeted FAP.

[0394] Application Example 2

[0395] [ 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:

[0396] HT1080-FAP cells were prepared at 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,

[0397] [ 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 solution and incubated in a 37°C incubator. 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 taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count; for blocking experiments, excess unlabeled DOTA-FAPI-04 was used to incubate with the corresponding radiolabeled drugs. After 60 minutes of incubation, PBS solution was used to terminate its uptake, and the cells were lysed with 1M NaOH. The cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0398] like Fig. 20 As shown, in the application example 2 of the present invention, the in vitro HT1080-FAP cell uptake [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,

[0399] [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-02,

[0400] [ 177 Lu]Lu-DOTA-HBED-CC-PEG3-FAPI-04 and [ 177 Lu] Lu-DOTA-FAPI-04 uptake-time curve (n=3); Fig.21 As shown, in Example 2 of the present invention, the in vitro HT1080-FAP cells are [ 177 Lu]Lu-DOTA-HBED-CC-FAPI-02,[ 177 Lu]Lu-DOTA-HBED-CC-FAPI-04,[ 177Lu]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).

[0401] Application Example 3

[0402] [ 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:

[0403] HT1080-FAP cells were prepared at 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. The uptake was terminated with PBS solution, the cells were lysed with 1M NaOH, the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count; for blocking experiments, an excess of unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drugs. After 60 minutes of incubation, the uptake was terminated with PBS solution, the cells were lysed with 1M NaOH, the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0404] like Fig. 22 As shown, the in vitro HT1080-FAP cell uptake in Example 3 of the present invention is 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04, [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177Lu] Lu-DOTA-FAPI-04 uptake-time curve (n=3); Fig.23 As shown, in Example 3 of the present invention, the in vitro HT1080-FAP cells are [ 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).

[0405] The results of in vitro cell uptake experiments show that [ 177 Lu]Lu-AAZTA-FAPI-04, [ 177 Lu]Lu-AAZTA-NI-FAPI-04,

[0406] [ 177 Lu]Lu-DOTA-NI-FAPI-04 and [ 177 The 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 similar to [ 177 Lu]Lu-DOTA-FAPI-04 is similar or slightly higher than [ 177 Lu]Lu-DOTA-FAPI-04.

[0407] Application Example 4

[0408] [ 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:

[0409] HT1080-FAP cells were prepared to 4 × 10 5500 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. The uptake was terminated with PBS solution, the cells were lysed with 1M NaOH, the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count; for blocking experiments, an excess of unlabeled DOTA-FAPI-04 was incubated with the corresponding radiolabeled drugs. After 60 minutes of incubation, the uptake was terminated with PBS solution, the cells were lysed with 1M NaOH, the cell lysate was taken up with filter paper and plugged into a plastic test tube to determine the radioactivity count.

[0410] like Fig.24 As shown, the in vitro HT1080-FAP cell uptake in Example 4 of the present invention is 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); Fig.25 As shown, in Example 4 of the present invention, the in vitro HT1080-FAP cells are [ 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).

[0411] The results of in vitro cell uptake experiments show that [ 68 Ga]Ga-HBED-CC-NI-FAPI-04, [ 68 Ga]Ga-NI-HBED-CC-FAPI-04,

[0412] [ 68The 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,

[0413] [ 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 [ 68 Ga]Ga-DOTA-FAPI-04.

[0414] Application Example 5

[0415] [ 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 imaging of U87MG tumor-bearing mice with Ga]Ga-DOTA-FAPI-04.

[0416] 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-[ 68Ga]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 evenly with an excess of DOTA-FAPI-04 and then injected through the tail vein of tumor-bearing mice, and imaging was performed at 60 minutes.

[0417] like Fig.26 As shown, in the application example 4 of the present invention, [ 68 Ga]Ga-AAZTA-FAPI-04, [ 68 Ga]Ga-AAZTA-NI-FAPI-04,

[0418] [ 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 imaging results of U87MG tumor-bearing mice with Ga]Ga-DOTA-FAPI-04.

[0419] The results of PET / CT imaging in mice bearing U87MG tumors showed that all compounds were taken up in the tumor site, and the blocking experiment showed that the 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 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.

[0420] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A radiolabeled FAPI complex, the general structural formula of which is shown below: in, 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 for chelating radionuclides, 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+ , 212 Bi 3+ wait; L1 and L2 are linking groups, wherein L1 is selected from any one of the following: Wherein, n is an integer from 0 to 6; L2 is selected from any of the following: A is the regulatory atom of the FAP targeting group, and A is either an H atom or a F atom.

2. The radiolabeled FAPI complex according to claim 1, wherein the specific structure is:

3. A preparation of a radiolabeled 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, and tert-butyl N-(3-bromopropyl)carbamate is added. After the reaction is completed, the mixture is filtered and extracted, and the organic phase is dried and purified to obtain a yellow-green oily compound, which is then deprotected with trifluoroacetic acid to obtain a white solid intermediate 1; (2) (S)-4-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid was dissolved in ultra-dry N,N-dimethylformamide, and 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-dihydro ... 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-cyanopyrrolidin-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); (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)carbamyl tert-butyl)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate, (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 dried 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(CH2CH2O) was added under ice bath. n CH2CH2Br), reflux reaction, overnight, suction filtration, distillation of solvent, purification, to obtain light yellow oily intermediate 3 (PEG n -FAPI); (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, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, stirred, and a white solid substance was added. After the reaction was completed, extraction was performed, the organic phase was dried, and purified to obtain a colorless solid intermediate 4 (Chelator-HBED-CC(tBu)2); (5) dissolving the chelating agent in ultra-dry N,N-dimethylformamide, adding 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under ice bath, stirring, adding intermediate 2 prepared in step (2) or FAPI after deprotection of Boc 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-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, extraction, drying the organic phase, and purification to obtain a light yellow oil, deprotection with trifluoroacetic acid, and purification by semi-preparative HPLC to obtain Chelator-NI-FAPI or Chelator-FAPI labeled precursor; (6) 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 1 prepared in step (1) is added, and the reaction is carried out at room temperature overnight. After the reaction is completed, extraction is performed, and the organic phase is 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 are added under ice bath. 7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine, 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, extract after the reaction, dry the organic phase, purify to obtain a light yellow oil, deprotect with trifluoroacetic acid, purify by semi-preparative HPLC to obtain NI-Chelator-FAPI labeled precursor; (7) The intermediate 4 prepared in step (4) is dissolved in N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are 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-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) or intermediate 3 prepared in step (3), after the reaction, extraction, drying the organic phase, purification, to obtain a light yellow oily substance, deprotection with trifluoroacetic acid, purification by semi-preparative HPLC, to obtain Chelator-HBED-CC-(PEG) n - FAPI labeled precursors; (8) The labeled precursor obtained in steps (5), (6) and (7) is dissolved in dimethyl sulfoxide, a sodium acetate buffer solution and a solution containing a radioactive nuclide are added, and the mixture is heated to obtain a corresponding radioactive labeled product.

4. The method for preparing a radiolabeled FAPI complex according to claim 3, characterized in that: In step (1), the nitroimidazole is added in an amount of 1 equivalent; the anhydrous potassium carbonate is added in an amount of 3-5 equivalents; the ultra-dry N,N-dimethylformamide is added in an amount of 10-20 mL; the N-(3-bromopropyl)carbamic acid tert-butyl ester is added in an amount of 1-2 equivalents; filtration is performed using diatomaceous earth, 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.

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-fluorene-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-azobenzotriazole)-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 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 (specifically including (S)-6-[3-(4-Boc-1-piperazinyl)propoxy]-N-[2-(2-cyano-4,4-difluoro-1-pyrrolidinyl)-2-oxygen) after deprotection of the Boc protecting group by trifluoroacetic acid 2-((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-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 - tert-butyl carboxylate, (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; extracted with ethyl acetate and saturated brine, the organic phase was dried over anhydrous sodium sulfate and purified by silica gel column chromatography; extracted with ethyl acetate and saturated brine, the organic phase was dried over anhydrous sodium sulfate and purified by speed purification chromatograph.

6. The method for preparing the radiolabeled FAPI complex according to claim 5, characterized in that: In step (3), the reaction is refluxed at 80° C.; potassium carbonate is removed by diatomaceous earth filtration, the filtrate is evaporated to remove the solvent under reduced pressure, and the residue is purified by flash purification chromatography; 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) 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; 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 of addition 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-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)acetic acid Cyclododecane-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-tetraazacyclododecane-1-yl)-5-tert-butoxy-5-oxopentanic 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-oxopentan-2-yl)-7-(1,5-di-tert-butoxy-1,5-dioxopentan-2-yl)-1,4,7,10-tetraazacyclododecane-1-yl)-5-tert-butoxy-5-oxopentanic acid (DOTA(GA)2(tBu)5), 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; the amount of the N-Cbz-1,2-diaminoethane added is 1.2-1.5 equivalents; the mixture is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and the spectral purification 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 10-20mL; the amount of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate added is 1 equivalent; the amount of N,N'-diisopropylethylamine added is 3 equivalents; a white solid substance is added, and after the reaction is completed, it 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).

8. The method for preparing the 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-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-tetraazacyclododecane-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-tetraazacyclododecane-1-yl)-5-tert-butoxy-5-oxopentanic acid (DOTA(GA)2(tBu)5), 2-(4,7-bis(2-(tert-butoxy)-2-oxopentan-2-yl)-1,4,7,10-tetraazacyclododecane-1-yl)pentanoic acid (DOTAGA(tBu)4), 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 trifluoroethyl FAPI after acid removal of Boc protecting group (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) were added in an amount of 1 equivalent; after the reaction was completed, 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 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))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); the chelating agent is added in an amount of 1 equivalent; the ultra-dry N,N -dimethylformamide is added in an amount of 2-10mL; the 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate is added in an amount of 1 equivalent; the N,N'-diisopropylethylamine is added in an amount of 3-5 equivalents; the intermediate 1 is added in an amount of 1 equivalent; after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified by a speed purification chromatograph; the light yellow oil, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate, N,N'-diisopropylethylamine 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)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- The amount of 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-carboxylate) added is 1 equivalent; the amount of ultra-dry N,N-dimethylformamide added is 7-10mL; after the reaction is completed, it is extracted with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified by a speed purification chromatograph; 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 deprotection of Boc 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-cyanopyrrolidinyl)-2-oxoethyl]quinoline-4-carboxamide, (1S,4S)-5-(3-((4-((2-((S)-2-cyano-4,4-difluoropyrrolidyl)-2-oxoethyl]quinoline-4-carboxamide, pyrrolidin-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 added in an amount of 1-1.5 equivalents; in step (7), after the reaction is completed, extraction is performed with ethyl acetate and saturated brine, the organic phase is dried over anhydrous sodium sulfate, and purified by a rapid purification chromatograph; in step (8), the solution containing the radionuclide is [. 68 Ga]GaCl3, [ 177 Lu]LuCl3, [ 18 F]AlF.

10. Use of the radiolabeled FAPI complex according to any one of claims 1 to 2 in the preparation of targeted radioactive diagnostic and therapeutic drugs.

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