Targeted FAP technetium-labeled compound and application thereof

Abstract

The invention discloses a technetium-labeled compound targeting FAP and application thereof, and belongs to the technical field of radiopharmaceutical chemistry and clinical nuclear medicine. The invention provides an FAPI04 or FAPI46 derivative containing a 6-hydrazino nicotinamide group, and the FAPI04 or FAPI46 derivative containing the 6-hydrazino nicotinamide group comprises the FAPI04 or the FAPI46 and the 6-hydrazino nicotinamide group which is connected with the FAPI04 or the FAPI46 through a connecting agent. The invention further provides a radionuclide M labeled FAPI04 or FAPI46 derivative, and the radionuclide M labeled FAPI04 or FAPI46 derivative is formed by coordination of the 6-hydrazino nicotinamide group-containing FAPI04 or FAPI46 derivative, a co-ligand and a radionuclide. The radioactive preparation is simple and convenient in preparation method, high in radiochemical purity and good in stability, has very high uptake and high target-to-non-target ratio at tumor parts of tumor-bearing mice, has specific binding with FAP in tumors, and is a novel tumor radioactive drug with popularization and application values.

Description

Technical Field

[0001] The present invention belongs to the technical fields of radiopharmaceutical chemistry and clinical nuclear medicine, and particularly relates to a technetium-labeled compound targeting FAP and application thereof. Background Art

[0002] Fibroblast activation protein (FAP) is highly expressed in most epithelial cell-derived tumor-associated fibroblasts, but is lowly expressed in normal tissues and organs. It is closely related to tumor migration, angiogenesis, immune surveillance escape, and other activities. Therefore, radionuclide-labeled fibroblast activation protein inhibitor (FAPI) and its derivatives are promising cancer diagnosis and treatment tracers that have attracted extensive attention and research. Based on the continuous modification and improvement of the structure of FAPI, a series of 68 Ga and 18 F-labeled FAPI complexes have been reported to be used for early diagnostic imaging of tumors such as colorectal cancer, pancreatic cancer, ovarian cancer, and melanoma. However, they are all PET imaging agents and are expensive, and their clinical application and promotion are subject to certain limitations. 99m Tc is the most common SPECT imaging radionuclide. 99 Mo / 99m Tc generator elution is obtained. 99m Tc-labeled drugs can be prepared in kit form and are easy to be promoted and used clinically. Therefore, a new drug targeting FAP is developed. 99m Tc tumor radiopharmaceuticals have important practical significance.

[0003] at present, 99m There are few reports on Tc-labeled molecular probes targeting FAP tumors, such as the published [ 99m Tc]Tc-FAPI-34 and [ 99m Tc][Tc-(CN-PEG4-FAPI)6] + It has high affinity and high tumor uptake ability, but the high uptake of non-target organs in the abdomen restricts its application to a certain extent. Patent application number ZL2021115355016 discloses a D-proline-modified FAPI derivative containing a HYNIC group (HYNIC-DP-FAPI) and a co-ligand tricine and TPPTS. 99m Tc coordinates to form a stable [ 99mTc]-Tc-HYNIC-DP-FAPI)(tricine / TPPTS). This complex exhibits good in vitro and in vivo stability and FAP affinity. Biodistribution studies show high uptake in tumors of tumor-bearing mice, with lower uptake in non-target organs such as the heart, liver, and muscle. However, due to high blood uptake, the tumor-to-blood ratio needs to be improved.

[0004] The linker connects the targeting group and the chelating group connected to the radionuclide, and plays an important role in regulating the affinity and biodistribution performance of radiopharmaceuticals in the body. 99m The tumor uptake and tumor / blood ratio of Tc-labeled FAP tumor molecular probes need to be improved. The present invention modifies the pharmacophore of FAPI04 or FAPI46 to obtain ligands containing different linkers, and the co-ligands (tricine / TPPTS or tricine / EDDA), and 99m Tc coordination forms a stable FAP-targeted tumor molecular probe with high preparation stability, strong targeting, good target-to-non-target ratio and easy promotion, which can be used for early diagnosis, staging and efficacy evaluation of tumors, and will lay a good foundation for the realization of translational medicine and precision medicine. Summary of the Invention

[0005] In order to overcome the defects in the above-mentioned prior art 99m To address the challenges of improving tumor uptake and tumor-to-blood ratios of Tc-labeled FAP-targeting tumor molecular probes, the present invention provides 6-hydrazinonicotinamide-containing FAPI04 or FAPI46 derivatives and their applications. The FAP pharmacophore (FAPI04 or FAPI46) and a metal ligand (6-hydrazinonicotinamide, represented by the hydrazone formed by the condensation of sodium benzaldehyde-2-sulfonate and 6-hydrazinonicotinamide) are linked using groups such as glycine-L-allylglycine, glycine-L-allylglycine-polyethylene glycol (PEGn), or proline-PEGn as linkers to prepare 6-hydrazinonicotinamide-containing FAPI04 or FAPI46 derivatives. Subsequently, multiple radiolabeled derivatives are designed and synthesized using tricine and TPPTS, or tricine and EDDA, ​​as co-ligands (i.e., tricine / TPPTS or tricine / EDDA). These derivatives are used for the diagnosis or treatment of diseases characterized by FAP overexpression, exploring their potential in tumor diagnosis and treatment.

[0006] This radiolabeled derivative has good stability and is easy to prepare. After being radiolabeled, it can be used for tumor diagnosis and treatment. It has high tumor uptake and a good target / non-target ratio, and has important scientific significance and application prospects in the field of tumor diagnosis and treatment.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] In one aspect, the present invention provides a FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group, comprising FAPI04 or FAPI46, and a 6-hydrazinonicotinamide group connected by a linker;

[0009] Preferably, the linker is glycine-L-allylglycine, glycine-L-allylglycine-polyethylene glycol or proline-polyethylene glycol.

[0010] The FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group has a general structural formula as shown in the following formula (I):

[0011]

[0012] Among them, when R 1 When it is O, X is selected from

[0013] When R 1 for When X is selected from

[0014] n is any integer from 1 to 6.

[0015] In a second aspect, the present invention provides the use of the aforementioned FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group in the preparation of a drug for treating cancer.

[0016] In a third aspect, the present invention provides a FAPI04 or FAPI46 derivative labeled with a radionuclide M, which is formed by the coordination of the FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group, a co-ligand and the radionuclide M.

[0017] The FAPI04 or FAPI46 derivative labeled with radionuclide M, the co-ligand is tris(hydroxymethyl)glycine and sodium triphenylphosphine trisulfonate, or tricine and ethylenediamine-N,N'-diacetic acid.

[0018] The FAPI04 or FAPI46 derivative labeled with a radionuclide M, wherein the radionuclide M is a metal radionuclide;

[0019] Preferably, the radionuclide M is 99m Tc, 99 Tc, 94m Tc,94 Tc, 52 Mn, 186 Re or 188 Re;

[0020] More preferably, the radionuclide M is 99m Tc.

[0021] The FAPI04 or FAPI46 derivative labeled with a radionuclide M, the structural formula of the FAPI04 or FAPI46 derivative labeled with a radionuclide M is shown in the following formula (II) or (III):

[0022]

[0023] Among them, when R 1 When it is O, X is selected from

[0024] When R 1 for When X is selected from

[0025] n is any integer from 1 to 6.

[0026] In a fourth aspect, the present invention provides a tumor imaging agent or an anti-tumor radiopharmaceutical comprising a FAPI04 or FAPI46 derivative labeled with any one of the radionuclide M described above.

[0027] In a fifth aspect, the present invention provides the use of any one of the radionuclide M-labeled FAPI04 or FAPI46 derivatives in the preparation of a tumor imaging agent.

[0028] In a sixth aspect, the present invention provides the use of any one of the radionuclide M-labeled FAPI04 or FAPI46 derivatives in the preparation of anti-tumor drugs.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The present invention provides a FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group, which is labeled with a radioactive nuclide to obtain a radioactive preparation. The preparation method is simple, the radiochemical purity is high, the stability is good, and the uptake and target-to-non-target ratio at the tumor site of tumor-bearing mice are high. In addition, the preparation has specific binding to FAP in the tumor, and the preparation is a new tumor radiopharmaceutical with promotion and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 Synthesis flow chart of intermediate FAPI04 or FAPI46-PEGn-NH2 series compounds;

[0032] Figure 2 Synthesis flow chart of the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6;

[0033] Figure 3 Synthesis flow chart of the labeled precursor VI-3-OSu-D / L-Pro-FAPI04 / 46-PEGn series compounds;

[0034] Figure 4 Flowchart of the synthesis of the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46;

[0035] Figure 5 Labeling roadmap for radiopharmaceuticals;

[0036] Figure 6 Radiopharmaceuticals 99m HPLC analysis chromatogram of Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPTS;

[0037] Figure 7 Radiopharmaceuticals 99m HPLC analysis chromatogram of Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI46-PEG2-tricine_TPPTS;

[0038] Figure 8 Radiopharmaceuticals 99m HPLC analysis chromatogram of Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_TPPTS;

[0039] Figure 9 Radiopharmaceuticals 99m HPLC analysis chromatogram of Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_EDDA;

[0040] Figure 10 Radiopharmaceuticals 99m HPLC analysis chromatogram of Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI04-PEG6-tricine_TPPTS;

[0041] Figure 11 [ 99mIn vitro stability HPLC chromatograms of [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPTS, wherein (A) is the in vitro stability HPLC chromatogram of physiological saline at room temperature for 6 hours, and (B) is the in vitro stability HPLC chromatogram of mouse serum at 37°C for 6 hours;

[0042] Figure 12 [ 99m In vitro stability HPLC chromatograms of [Tc]Tc-HYNIC-D-Pro-VI-8-de_Fmoc-FAPI46-PEG2-tricine_TPPTS, wherein (A) is the in vitro stability HPLC chromatogram of normal saline at room temperature for 6 hours, and (B) is the in vitro stability HPLC chromatogram of mouse serum at 37°C for 6 hours;

[0043] Figure 13 [ 99m In vitro stability HPLC chromatograms of [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_TPPTS, wherein (A) is the in vitro stability HPLC chromatogram of physiological saline at room temperature for 6 hours, and (B) is the in vitro stability HPLC chromatogram of mouse serum at 37°C for 6 hours;

[0044] Figure 14 [ 99m In vitro stability HPLC chromatograms of [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_EDDA, ​​wherein (A) is the in vitro stability HPLC chromatogram of physiological saline at room temperature for 6 hours, and (B) is the in vitro stability HPLC chromatogram of mouse serum at 37°C for 6 hours;

[0045] Figure 15 [ 99m Figure 2 shows the in vitro stability HPLC chromatogram of Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI04-PEG6-tricine_TPPTS, where (A) is the in vitro stability HPLC chromatogram of physiological saline at room temperature for 6 h, and (B) is the in vitro stability HPLC chromatogram of mouse serum at 37°C for 6 h. DETAILED DESCRIPTION

[0046] To make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below. However, it should be understood that the description herein is only for the purpose of explaining the present invention and is not intended to limit the scope of the present invention.

[0047] Unless otherwise defined, all technical and scientific terms herein have the same meanings as those commonly understood by those skilled in the art to which the present invention pertains. The terms used in this specification are intended only to describe specific embodiments and are not intended to limit the present invention. The reagents and instruments used herein are commercially available. The characterization methods involved can be found in the relevant descriptions in the prior art and will not be detailed here.

[0048] Example 1: Synthesis of intermediate FAPI04 or FAPI46-PEGn-NH2 series compounds

[0049] The synthetic flow chart of the intermediate FAPI04 or FAPI46-PEGn-NH2 series compounds is as follows: Figure 1 shown.

[0050] 1) Synthesis of intermediate FAPI04-PEG2-NHBoc

[0051] The starting material FAPI04 (100 mg, 0.1665 mmol, 1.2000 equiv.) and PEG2-NHBoc-CO2H (38.5 mg, 0.1387 mmol, 1.0000 equiv.) were dissolved in ultra-dry DMF (0.77 mL). N-methylimidazole (NMI, 35.3 mg, 0.4299 mmol, 3.1000 equiv.) was added, followed by the addition of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (TCFH, 42.8 mg, 0.1526 mmol, 1.1000 equiv.) in portions. The mixture was stirred at room temperature for 1 hour. After the reaction was nearly complete as monitored by TLC, water (1 mL) was added to the mixture, which was then concentrated under reduced pressure and purified by silica gel column chromatography to afford the intermediate FAPI04-PEG2-NHBoc (101.4 mg, 97.93% yield). 1H NMR(400MHz, Methanol-d4)δ8.74(d,J=4.4Hz,1H),7.97–7.94(m,2H),7.58–7.56(m,2H),7.45(dd ,J1=9.2Hz,J2=2.6Hz,1H),7.06(s,0.5H),6.94(s,0.5H),5.12(dd,J1=9.4Hz,J2=2.8Hz,1H),4.3 5–4.08(m,6H),3.76–3.73(m,4H),3.63–3.59(m,8H),3.49(t,J=5.6Hz,2H),3.21(t,J=5.6Hz,2H) ,2.68–2.62(m,2H),2.55(t,J=4.6Hz,2H),2.49(t,J=5.0Hz,2H),2.12–2.04(m,2H),1.42(s,9H); 19 F NMR(376MHz, Methanol-d4)δ-73.93,-75.81,-97.80,-98.42,-105.45,-106.08.

[0052] 2) Synthesis of the intermediate FAPI46-PEG2-NHBoc

[0053] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The raw material FAPI46 (100 mg, 0.2002 mmol, 1.2000 equiv.) was reacted with PEG2-NHBoc-CO2H (46.3 mg, 0.3503 mmol, 1.0000 equiv.) to obtain the intermediate FAPI46-PEG2-NHBoc (123.5 mg, yield 97.48%). 1 H NMR(400MHz, Methanol-d4)δ8.52(d,J=4.4Hz,1H),7.87(d,J=9.4Hz,1H),7.59–7.53 (m,2H),7.46–7.41(m,2H),7.06(s,0.5H),6.94(s,0.5H),5.13–5.10(m,1H),4.35–4. 08(m,4H),3.75–3.72(m,4H),3.65–3.57(m,10H),3.48–3.45(m,2H),3.20–3.17(m,2H ),3.12(s,3H),2.66–2.63(m,2H),2.49–2.44(m,6H),1.91–1.86(m,2H),1.41(s,9H); 19F NMR(376MHz, Methanol-d4)δ-73.93,-75.81,-97.80,-98.42,-105.45,-106.08.

[0054] 3) Synthesis of intermediate FAPI04-PEG4-NHBoc

[0055] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The raw material FAPI04 (150 mg, 0.2498 mmol, 1.2000 equiv.) was reacted with PEG4-NHBoc-CO2H (76.1 mg, 0.2081 mmol, 1.0000 equiv.) to obtain the intermediate FAPI46-PEG4-NHBoc (156.9 mg, yield 90.35%). 1 H NMR(400MHz, Methanol-d4)δ8.75(d,J=4.4Hz,1H),7.98–7.96(m,2H),7.60(s,1H),7.57(d,J =4.4Hz,1H),7.47–7.44(m,1H),7.07(s,0.5H),6.94(s,0.5H),5.14–5.11(m,1H),4.31–4.09( m,6H),3.76–3.73(m,4H),3.62–3.58(m,16H),3.52(t,J=5.5Hz,2H),3.24(t,J=5.3Hz,2H),2. 70–2.63(m,4H),2.56(t,J=4.6Hz,2H),2.50(t,J=4.8Hz,2H),2.13–2.06(m,2H),1.43(s,9H); 19 F NMR(376MHz, Methanol-d4)δ-73.93,-75.81,-98.13,-98.76,-105.34,-105.97.

[0056] 4) Synthesis of the intermediate FAPI46-PEG4-NHBoc

[0057] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The raw material FAPI46 (50 mg, 0.1001 mmol, 1.2000 equiv.) was reacted with PEG4-NHBoc-CO2H (30.5 mg, 0.2081 mmol, 1.0000 equiv.) to obtain the intermediate FAPI46-PEG4-NHBoc (93.0 mg). 1H NMR(400MHz,Methanol-d4)δ8.52(d,J=4.4Hz,1H),7.87(d,J=9.4Hz,1H),7.59(s,1H) ,7.57–7.54(m,1H),7.48–7.42(m,2H),7.07(s,0.5H),6.94(s,0.5H),5.14–5.11(m,1 H),4.29–4.09(m,4H),3.75–3.72(m,4H),3.62–3.55(m,20H),3.26(t,J=5.3Hz,2H),3 .12(s,3H),2.69(t,J=5.6Hz,2H),2.48–2.42(m,6H),1.91–1.86(m,2H),1.42(s,9H); 19 F NMR(376MHz, Methanol-d4)δ-74.01,-75.89,-97.80,-98.42,-105.46,-106.08.

[0058] 5) Synthesis of intermediate FAPI04-PEG6-NHBoc

[0059] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The raw material FAPI04 (100.0 mg, 0.1665 mmol, 1.2000 equiv.) was reacted with PEG6-NHBoc-CO2H (62.9 mg, 0.1388 mmol, 1.0000 equiv.) to obtain the intermediate FAPI46-PEG4-NHBoc (115.0 mg, yield 89.89%). 1 H NMR(400MHz, Methanol-d4)δ8.75(d,J=4.4Hz,1H),7.98–7.95(m,2H),7.62(s,1H),7.57( d,J=4.4Hz,1H),7.47–7.44(m,1H),7.08(s,0.5H),6.96(s,0.5H),5.14–5.11(m,1H),4.31 –4.08(m,6H),3.74–3.73(m,4H),3.67–3.59(m,24H),3.52–3.49(m,2H),3.23–3.21(m,2H) ,2.70–2.64(m,4H),2.58–2.56(m,2H),2.53–2.50(m,2H),2.13–2.07(m,2H),1.43(s,9H); 19FNMR(376MHz, Methanol-d4)δ-73.87,-75.75,-98.01,-98.64,-105.46,-106.08.

[0060] 6) Synthesis of the intermediate FAPI46-PEG6-NHBoc

[0061] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The raw material FAPI46 (50.0 mg, 0.1001 mmol, 1.2000 equiv.) was reacted with PEG6-NHBoc-CO2H (37.8 mg, 0.0834 mmol, 1.0000 equiv.) to obtain the intermediate FAPI46-PEG2-NHBoc (74.4 mg, yield 95.42%). 1 H NMR(400MHz, Methanol-d4)δ8.52(d,J=4.4Hz,1H),7.89–7.86(m,1H),7.58(s,1H),7.57–7 .54(m,1H),7.46–7.45(m,2H),7.07(s,0.5H),6.94(s,0.5H),5.14–5.11(m,1H),4.28–4.0 9(m,4H),3.73–3.72(m,4H),3.64–3.60(m,26H),3.49(t,J=5.6Hz,2H),3.21(t,J=5.5Hz,2 H),3.12(s,3H),2.67(t,J=5.9Hz,2H),2.49–2.42(m,6H),1.91–1.86(m,2H),1.43(s,9H); 19 FNMR(376MHz, Methanol-d4)δ-74.01,-75.89,-97.80,-98.42,-105.46,-106.08.

[0062] 7) Synthesis of intermediate FAPI04-PEG2-NH2

[0063] A solution of the intermediate FAPI04-PEG2-NHBoc (101.4 mg, 0.1360 mmol) in ultra-dry dichloromethane (1.1 mL) was cooled to 0°C, and trifluoroacetic acid (494.7 mg, 31.9153 equiv.) was added dropwise. The mixture was then returned to room temperature and stirred for 1.5 h. After completion of the reaction, methanol (1.1 mL) was added incrementally, and the solvent was removed by concentration under reduced pressure using a circulating water vacuum pump. This process was repeated 6-9 times to obtain the intermediate FAPI04-PEG2-NH2 (103.3 mg). This product was used directly in the next reaction without purification.

[0064] 8) Synthesis of intermediate FAPI46-PEG2-NH2

[0065] The synthesis procedure was similar to that in Example 1, "7) Synthesis of Intermediate FAPI04-PEG2-NH2." Intermediate FAPI46-PEG2-NHBoc (123.5 mg, 0.1627 mmol) was used to obtain intermediate FAPI46-PEG2-NH2 (156.6 mg). This product was used directly in the next reaction without purification.

[0066] 9) Synthesis of intermediate FAPI04-PEG4-NH2

[0067] The synthesis procedure was similar to that in Example 1, "7) Synthesis of Intermediate FAPI04-PEG2-NH2." Intermediate FAPI04-PEG4-NHBoc (156.9 mg, 0.1881 mmol) was used to obtain intermediate FAPI04-PEG4-NH2 (159.5 mg). This product was used directly in the next reaction without purification.

[0068] 10) Synthesis of intermediate FAPI46-PEG4-NH2

[0069] The synthesis procedure was similar to that in Example 1, "7) Synthesis of Intermediate FAPI04-PEG2-NH2." Intermediate FAPI46-PEG4-NHBoc (93.0 mg, 0.1098 mmol) was used to obtain intermediate FAPI46-PEG4-NH2 (94.5 mg). This product was used directly in the next reaction without purification.

[0070] 11) Synthesis of intermediate FAPI04-PEG6-NH2

[0071] The synthesis procedure was similar to that in Example 1, "7) Synthesis of Intermediate FAPI04-PEG2-NH2." Intermediate FAPI04-PEG4-NHBoc (115.0 mg, 0.1245 mmol) was used to obtain intermediate FAPI04-PEG6-NH2 (116.7 mg). This product was used directly in the next reaction without purification.

[0072] 12) Synthesis of intermediate FAPI46-PEG6-NH2

[0073] The synthesis procedure was similar to that in Example 1, "7) Synthesis of Intermediate FAPI04-PEG2-NH2." Intermediate FAPI46-PEG6-NHBoc (74.4 mg, 0.0796 mmol) was used to obtain intermediate FAPI46-PEG6-NH2 (75.5 mg). This product was used directly in the next reaction without purification.

[0074] Example 2: Synthesis of Labeling Precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6

[0075] The flow chart of the synthesis of the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6 is as follows Figure 2 shown.

[0076] 1) Synthesis of intermediate VI-3-OSu

[0077] To a suspension of compound VI-1 (5.0000 g, 32.6499 mmol, 1.0000 equiv.) in ultra-dry DMF (201 mL) was added compound VI-2 (7.2147 g, 34.6578 mmol, 1.0615 equiv.), and the mixture was stirred at room temperature for 3 h. The mixture was then cooled to 0°C, and N-hydroxysuccinimide (NHS, 3.7577 g, 32.6499 mmol, 1.0000 equiv.) was added, followed by the dropwise addition of N,N'-dicyclohexylcarbodiimide (DCC, 13.8876 g, 67.3077 mmol, 2.0615 equiv.). The mixture was then allowed to return to room temperature and stirred for 18 h. After the reaction was essentially complete as monitored by TLC, the mixture was filtered under reduced pressure, and the filtrate was concentrated under reduced pressure using a circulating water vacuum pump. After ethyl acetate (201 mL) was added and refluxed, the mixture was filtered under reduced pressure to obtain intermediate VI-3-OSu (13.2744 g, light yellow solid, yield 92.33%). 1H NMR (400MHz, DMSO-d6) δ11.91(s,1H),9.16(s,1H),8.79(d,J=1.9Hz,1H),8.18–8 .15(m,1H),8.05–8.03(m,1H),7.80–7.78(m,1H),7.39–7.36(m,3H),2.89(s,4H).

[0078] 2) Synthesis of intermediate VI-7

[0079] The synthesis steps were as described in "1) Synthesis of intermediate VI-3-OSu" in Example 2. Intermediate VI-5 (5.0000 g, 13.2582 mmol, 1.0000 equiv.) was reacted with VI-6 (2.0857 g, 15.9098 mmol, 1.2000 equiv.) to obtain intermediate VI-7 (3.6928 g, white solid, yield 55.30%). 1 H NMR(400MHz,Chloroform-d)δ7.75(d,J=7.5Hz,2H),7.57(d,J=7.3Hz,2H),7.40(t,J=7.4Hz,2H),7.32–7.28(m,2H),6.63(s,1H),5.77–5.7 3(m,1H),5.48–5.46(m,1H),5.17–5.13(m,2H),4.45–4.31(m,3H),4.2 1(t,J=6.9Hz,1H),3.98–3.86(m,2H),2.62–2.48(m,2H),1.45(s,9H).

[0080] 3) Synthesis of Compound VI-8

[0081] Intermediate VI-7 (3.6928 g, 8.1964 mmol, 1.0000 equiv.) and trimethylsilane (2.3827 g, 20.4910 mmol, 2.5000 equiv.) were dissolved in ultra-dry dichloromethane (17 mL). The mixture was cooled to 0°C, and trifluoroacetic acid (12.1492 g, 106.5530 mmol, 13.0000 equiv.) was added dropwise. The mixture was stirred at room temperature for 4 h. After the reaction was nearly complete as monitored by TLC, methanol (12 mL) was added incrementally, and the solvent was removed by concentration under reduced pressure using a circulating water vacuum pump. This process was repeated 6-9 times. Intermediate VI-8 was then purified by silica gel column chromatography. 1H NMR(400MHz, Methanol-d4)δ7.78(d,J=7.6Hz,2H),7.67–7.64(m,2H),7.38(t,J=7.4Hz,2H),7.32–7.28(m,2H),5.85–5.75(m,1H),5 .16–5.07(m,2H),4.43–4.38(m,1H),4.35–4.31(m,1H),4.23–4.20(m,2H),3.96–3.85(m,2H),2.62–2.55(m,1H),2.42–2.35(m,1H).

[0082] 4) Synthesis of intermediate VI-8-de_Fmoc

[0083] Compound VI-8 (671.7 mg, 1.7037 mmol, 1.0000 equiv.) and octanethiol (2.4913 g, 17.0300 mmol, 10.0000 equiv.) were dissolved in ultra-dry DMF (17 mL). Under nitrogen, tetra(n-butyl)ammonium fluoride trihydrate (1.0746 g, 3.4060 mmol, 2.0000 equiv.) was added. The mixture was ultrasonically stirred at room temperature for 1 min. After the reaction was nearly complete as monitored by TLC, the solvent was removed by concentration under reduced pressure. The product was then initially isolated and purified by silica gel column chromatography to obtain intermediate VI-8-de_Fmoc. 1 H NMR (400MHz, Methanol-d4) δ5.74–5.66(m,1H),5.08–5.01(m,2H),3.73–3.58(m,2H),3.41–3.38(m,1H),2.44–2.38(m,1H),2.30–2.22(m,1H).

[0084] 5) Synthesis of intermediate VI-3-OSu-VI-8-de_Fmoc

[0085] The preliminarily isolated intermediate VI-8-de-Fmoc (314.3 mg, 1.5165 mmol, 1.0000 equiv.) and VI-3-OSu (667.8 mg, 1.8254 mmol, 1.2037 equiv.) were dissolved in ultra-dry DMF (17 mL). Triethylamine (886.3 mg, 8.7584 mmol, 5.7754 equiv.) was then added, and the mixture was stirred at room temperature for 24 hours. After the reaction was nearly complete, as monitored by TLC, the solvent was removed by concentration under reduced pressure, and the intermediate VI-3-OSu-VI-8-deFmoc (45.9 mg) was initially isolated and purified by silica gel column chromatography. 1H NMR (400MHz, Methanol-d4) δ8.67 (s, 1H), 8.40 (s, 1H), 8.08 (d, J = 7.8Hz, 1H), 7. 91(dd,J1=9.0Hz,J2=1.8Hz,1H),7.77(d,J=7.9Hz,1H),7.48(t,J=7.2Hz,1H),7. 37(t,J=7.7Hz,1H),7.08(d,J=9.0Hz,1H),5.79–5.69(m,1H),5.12–5.02(m,2H) ,4.52–4.49(m,1H),3.65(q,J=14.7Hz,2H),2.63–2.57(m,1H),2.49–2.41(m,1H)

[0086] 6) Synthesis of the Labeling Precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6

[0087] The synthesis was carried out by referring to step "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1, and compound VI-3-OSu-VI-8-de_Fmoc (14.7 mg, 0.0296 mmol, 1.0000 equiv.) was reacted with FAPI46-PEG6-NH2 (33.6 mg, 0.0355 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6 (15.2 mg, yield 39.13%). 1 H NMR (400MHz, Methanol-d4) δ8.99(s,1H),8.78(s,1H),8.52(d,J=4.1Hz,1H),8.22(d,J=7.7H z,1H),8.14(d,J=8.9Hz,1H),7.98–7.87(m,3H),7.50–7.34(m,7H),5.93–5.83(m,1H),5.23– 5.18(m,2H),5.12–5.09(m,1H),4.49–4.45(m,1H),4.33–4.06(m,6H),3.69–3.67(m,6H),3.5 8–3.52(m,26H),3.17–3.07(m,9H),2.73–2.69(m,2H),2.66–2.62(m,2H),2.11–1.98(m,4H); 19 F NMR(376MHz, Methanol-d4)δ-97.79,-98.41,-105.50,-106.12.

[0088] Example 3: Synthesis of Labeled Precursor VI-3-OSu-D / L-Pro-FAPI04 / 46-PEGn Series Compounds

[0089] The synthesis process of the labeled precursor VI-3-OSu-D / L-Pro-FAPI04 / 46-PEGn series compounds is as follows Figure 3 shown.

[0090] 1) Synthesis of intermediate VI-3-OSu-D-Pro

[0091] The synthesis procedure was similar to that of "Synthesis of Intermediate VI-3-OSu-VI-8-de_Fmoc" in Example 2. Intermediate VI-3-OSu (1 g, 2.2709 mmol, 1.0000 equiv.) was reacted with D-proline (314.7 mg, 2.7334 mmol, 1.2037 equiv.) to obtain intermediate VI-3-OSu-D-Pro (101.4 mg, 10.14% yield). 1 H NMR(400MHz,D2O)δ8.78(s,1H),8.20(s,0.5H),8.09(d,J=7.8Hz,1H),8.01(m,0.5H),7.97–7.95(m,0.5H),7.80–7.74(m,1.5H),7. 51–7.47(m,1H),7.42–7.37(m,1H),7.07–7.04(m,1H),4.29–4.20(m,1H),3.60–3.52(m,2H),2.27–2.14(m,1H),1.92–1.75(m,3H); 1 H NMR(400MHz, Methanol-d4)δ9.07(s,1H),8.37(s,0.8H),8.29–8.22(m,1.2H),8.04(d,J=8.6Hz,0.5H),7.99–7.88(m,1.50 H),7.50–7.40(m,2H),7.26–7.23(m,1H),4.57(t,J=12.6Hz,1H),3.74–3.72(m,2H),2.44–2.36(m,1H),2.07–1.94(m,3H).

[0092] 2) Synthesis of intermediate VI-3-OSu-L-Pro

[0093] The synthesis procedure was similar to that of "5) Synthesis of Intermediate VI-3-OSu-VI-8-de_Fmoc" in Example 2. Intermediate VI-3-OSu (500.0 mg, 1.1354 mmol, 1.0000 equiv.) was reacted with L-proline (157.4 mg, 1.3667 mmol, 1.2037 equiv.) to obtain intermediate VI-3-OSu-L-Pro (61.8 mg, 12.36% yield). 1 H NMR(400MHz,D2O)δ8.75–8.73(m,1H),8.18–8.16(m,0.59H),8.07–8.05( m,1H),7.80(s,0.5H),7.92–7.90(m,0.5H),7.74–7.67(m,1.5H),7.51–7 .47(m,1H),7.46–7.45(m,1H),7.37–7.31(m,1H),7.01–6.98(m,1H),4.2 6–4.21(m,1H),3.58–3.50(m,2H),2.24–2.14(m,1H),1.93–1.75(m,3H).

[0094] 3) Synthesis of Labeling Precursor VI-3-OSu-D-Pro-FAPI04-PEG2

[0095] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The intermediate VI-3-OSu-D-Pro (33.7 mg, 0.0765 mmol, 1.0000 equiv.) was reacted with FAPI04-PEG2-NH2 (69.8 mg, 0.0918 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-D-Pro-FAPI04-PEG2 (21.1 mg, yield 25.82%). 1 H NMR(400MHz,Methanol-d4)δ8.86–8.78(m,1H),8.66–8.58(m,1H),8.36(s,0.5H),8 .15–8.11(m,0.8H),8.02–8.00(m,0.7H),7.88–7.82(m,4H),7.63–7.11(m,7H),5.07 –5.05(m,1H),4.55–4.41(m,1H),4.22–3.97(m,4H),3.78–3.30(m,18H),2.85–2.81 (m,2H),2.76–2.47(m,8H),2.21–2.20(m,1H),2.11–2.04(m,2H),1.94–1.87(m,3H);19 F NMR(376MHz, Methanol-d4)δ-73.89,-75.77,-98.70,-105.44,-106.07.

[0096] 4) Synthesis of the Labeling Precursor VI-3-OSu-D-Pro-FAPI46-PEG2

[0097] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The intermediate VI-3-OSu-D-Pro (34.0 mg, 0.0772 mmol, 1.0000 equiv.) was reacted with FAPI46-PEG2-NH2 (71.6 mg, 0.0926 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-D-Pro-FAPI46-PEG2 (28.0 mg, 33.55% yield). 1 H NMR(400MHz,Methanol-d4)δ8.93(s,1H),8.52–8.48(m,2H),8.21–8.19(m ,1.4H),7.97–7.84(m,3H),7.69–7.67(m,0.6H),7.50–7.28(m,7H),5.13(d ,J=8.7Hz,1H),4.52–4.49(m,1H),4.31–4.06(m,4H),3.78–3.38(m,20H),3 .10–3.06(m,4H),2.69–2.53(m,9H),2.33–2.29(m,2H),2.05–1.87(m,2H).

[0098] 5) Synthesis of the Labeling Precursor VI-3-OSu-L-Pro-FAPI46-PEG4

[0099] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The intermediate VI-3-OSu-L-Pro (30.6 mg, 0.0695 mmol, 1.0000 equiv.) was reacted with FAPI46-PEG4-NH2 (71.8 mg, 0.0834 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-L-Pro-FAPI46-PEG4 (34.6 mg, yield 42.59%). 1H NMR(400MHz,Methanol-d4)δ8.95(s,1H),8.52(s,1H),8.21–8.19(m,1H),7 .98–7.86(m,3H),7.73–7.58(m,2H),7.50–7.20(m,5H),5.17–5.15(m,1H),4 .51–4.48(m,1H),4.30–4.12(m,4H),3.84–3.40(m,22H),3.16–2.97(m,9H), 2.89–2.78(m,6H),2.69–2.62(m,2H),2.31–2.07(m,2H),2.02–1.85(m,4H); 19 F NMR(376MHz, Methanol-d4)δ-73.79,-75.67,-76.85,-77.03,-97.84,-98.47,-105.42,-106.04.

[0100] 6) Synthesis of Labeling Precursor VI-3-OSu-D-Pro-FAPI04-PEG6

[0101] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The intermediate VI-3-OSu-D-Pro (33.7 mg, 0.0765 mmol, 1.0000 equiv.) was reacted with FAPI04-PEG6-NH2 (85.9 mg, 0.0918 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-D-Pro-FAPI04-PEG6 (30.1 mg, yield 31.61%). 1 H NMR(400MHz, Methanol-d4)δ8.91(s,1H),8.73(t,J=3.3Hz,1H),8.45(s,0.6H),8.24–8.19( m,2.4H),7.97–7.91(m,4H),7.56(s,1H),7.47–7.44(m,3H),7.38–7.21(m,4H),5.17–5.14( m,1H),4.51(t,J=7.2Hz,1H),4.32–4.09(m,6H),3.76–3.70(m,8H),3.62–3.56(m,24H),3.1 7–3.02(m,6H),2.67–2.65(m,2H),2.35–2.31(m,2H),2.21–2.17(m,3H),2.03–1.98(m,3H); 19F NMR(376MHz, Methanol-d4)δ-73.86,-75.74,-98.04,-98.66,-105.49,-106.12.

[0102] 7) Synthesis of Labeling Precursor VI-3-OSu-L-Pro-FAPI46-PEG6

[0103] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. The intermediate VI-3-OSu-L-Pro (16.2 mg, 0.0368 mmol, 1.0000 equiv.) was reacted with FAPI46-PEG6-NH2 (41.9 mg, 0.0442 mmol, 1.2000 equiv.) to obtain the labeled precursor VI-3-OSu-L-Pro-FAPI46-PEG6 (18.2 mg, 39.35% yield). 1 H NMR(400MHz,Methanol-d4)δ8.93–8.91(m,1H),8.53–8.46(m,1.5H),8.21– 8.11(m,1.8H),7.93–7.87(m,2.7H),7.51–7.21(m,8H),5.16–5.14(m,1H),4 .53–4.50(m,1H),4.33–4.07(m,6H),3.77–3.64(m,8H),3.56–3.48(m,24H), 3.15–2.99(m,9H),2.63–2.60(m,2H),2.35–2.23(m,2H),2.09–1.89(m,6H); 19 F NMR(376MHz, Methanol-d4)δ-76.90,-97.93,-97.95,-98.55,-98.58,-105.42,-106.05.

[0104] Example 4: Synthesis of Labeled Precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46

[0105] The synthesis process of the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46 is as follows Figure 4 shown.

[0106] 1) Synthesis of intermediate VI-8-FAPI46

[0107] The synthesis steps were as described in "1) Synthesis of intermediate FAPI04-PEG2-NHBoc" in Example 1. Intermediate VI-8 (227.7 mg, 0.1273 mmol, 1.0000 equiv.) was reacted with FAPI46 (425.1 mg, 0.6927 mmol, 1.2000 equiv.) to obtain intermediate VI-8-FAPI46 (345.6 mg, light yellow solid, yield 68.34%). 1 H NMR (400MHz, Methanol-d4) δ8.51(d,J=4.2Hz,1H),7.87(d,J=9.4Hz,1H),7.78(d,J=7.5Hz ,2H),7.65(d,J=7.3Hz,2H),7.54–7.51(m,1H),7.45(s,2H),7.38(t,J=7.4Hz,2H),7.30(t ,J=7.5Hz,2H),5.84–5.74(m,1H),5.15–5.06(m,3H),4.40–3.99(m,10H),3.67–3.53(m,4H ),3.52–3.49(m,2H),3.09(s,3H),2.95–2.76(m,2H),2.61–2.36(m,8H),1.89–1.82(m,2H); 19 FNMR(376MHz, Methanol-d4)δ-73.83,-73.96,-75.72,-75.84,-77.13,-97.99,-98.04,-98.60,-98.67,-105.24,-105.87.

[0108] 2) Synthesis of Compound VI-8-de_Fmoc-FAPI46

[0109] The synthesis steps refer to "4) Synthesis of intermediate VI-8-de_Fmoc" in Example 2, and intermediate VI-8-de_Fmoc-FAPI46 (123.8 mg, light yellow solid, yield 48.00%) was obtained from intermediate VI-8-FAPI46 (345.6 mg, 0.3945 mmol, 1.0000 equiv.). 1H NMR (400MHz, Methanol-d4) δ8.51(d,J=4.4Hz,1H),7.87(d,J=9.4Hz,1H),7.56–7.39(m,3H),5.86–5.75(m,1H),5.17–5.10(m,3H ),4.33–4.07(m,6H),3.68–3.55(m,4H),3.49–3.46(m,3H),3.11(s,3H),2.98–2.74(m,2H),2.52–2.32(m,8H),1.91–1.82(m,2H).

[0110] 3) Synthesis of the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46

[0111] The synthesis steps were as described in "5) Synthesis of intermediate VI-3-OSu-VI-8-de_Fmoc" in Example 2. The intermediate VI-8-de_Fmoc-FAPI46 (123.8 mg, 0.1894 mmol, 1.0000 equiv.) was reacted with VI-3-OSu (108.4 mg, 0.2462 mmol, 1.3000 equiv.) to obtain the labeled precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46 (24.0 mg, light yellow solid, yield 12.95%). 1 H NMR (400MHz, Methanol-d4) δ8.86(s,1H),8.54(d,J=1.9Hz,1H),8.42(d,J=4.4Hz,1H),8.13(d,J=7 .9Hz,1H),8.00–7.97(m,1H),7.84(d,J=7.7Hz,1H),7.78(d,J=9.4Hz,1H),7.44–7.19(m,10H),5.83 –5.73(m,1H),5.28–5.25(m,1H),5.06–5.04(m,2H),4.58–4.54(m,1H),4.18–3.93(m,5H),3.64–3. 47(m,7H),3.01(s,3H),2.69–2.58(m,6H),2.59–2.40(m,2H),2.23–2.15(m,2H),1.96–1.94(m,2H); 19 F NMR(376MHz, Methanol-d4)δ-75.72,-76.93,-98.02,-98.05,-98.65,-98.68,-105.31,-105.93,-105.94; HRMS(ESI+):m / z calculated for[C45 H 49 F2N 12 NaO8S+Na] + 1001.3275, found 1001.3346.

[0112] Example 5: Radiolabeling Experiment

[0113] 99m The labeling route of Tc-labeled radiopharmaceuticals is as follows Figure 5 shown.

[0114] 1) 99m Labeling method of Tc-labeled radiopharmaceuticals

[0115] For formula (II) (co-ligands are tricine and TPPTS), with radiopharmaceuticals [ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPPTS,[ 99m Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI46-PEG2-tricine_TPPPTS,[ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_TPPTS or [ 99m Take Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI04-PEG6-tricine_TPPTS as an example:

[0116] Weigh tricine (1 mg) and TPPTS (2 mg) and dissolve them in physiological saline (0.5 mL). Add succinate buffer (pH 5.0) to adjust the solution to pH 5.0. Then add the labeled precursor (10-20 μg) and freshly washed [ 99m [Tc]NaTcO4 in physiological saline solution (about 185-3700 MBq, 0.5 mL) and heated in a boiling water bath for 30 min. Alternatively, tricine (1 mg), TPPTS (2 mg), pH 5.0 succinate buffer, and labeling precursor (10-20 μg) can be prepared into a lyophilized kit and dissolved in physiological saline (0.5 mL). Freshly washed [ 99m[Tc] NaTcO4 in physiological saline (approximately 185-3700 MBq, 0.5 mL) was heated in a boiling water bath for 30 minutes. The reaction solution was cooled and transferred to a SepPak C18 Plus Light cartridge (Waters Corporation, pretreated with 10 mL of anhydrous ethanol and 10 mL of water). The cartridge was then washed with water (5 mL) and eluted with 1 mL of a mixture of ethanol and water (0.75 / 0.25, v / v). The eluate was purified using the HPLC separation method described below to obtain the radiopharmaceutical.

[0117] The labeling precursors corresponding to the above radioactive drugs are: VI-3-OSu-VI-8-de_Fmoc-FAPI46, VI-3-OSu-D-Pro-FAPI46-PEG2, VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6 or VI-3-OSu-D-Pro-FAPI04-PEG6.

[0118] HPLC semi-preparative column separation and purification conditions: Phase A: water (containing 0.1% trifluoroacetic acid), Phase B: acetonitrile (containing 0.1% trifluoroacetic acid), gradient: 10% Phase B (0-4 min), 10% to 90% Phase B (4-20 min), 90% to 10% Phase B (20-30 min), 90% to 10% Phase B (30-40 min), and 10% Phase B (40-60 min). Total flow rate: 2.363 mL / min. Beijing Huideyi QuikSep SP ODS-A reversed-phase C18 semi-preparative column, 10 μm particle size, 10 mm x 250 mm column.

[0119] 2) For formula (II) (co-ligands are tricine and EDDA), with radiopharmaceuticals [ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_EDDA as an example:

[0120] Weigh tricine (10 mg) and dissolve it in PBS buffer (pH 7.2, 0.05 M, 0.5 mL). Dissolve EDDA (5 mg) in NaOH aqueous solution (0.1 M, 0.25 mL). The above two solutions, the labeling precursor VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6 (39.6 μg) and freshly washed [ 99mTc]NaTcO4 in saline (about 185-3700 MBq, 0.5 mL), and SnCl2 (1 mg / mL in 0.1 M HCl, 20 μL) were added and heated in a boiling water bath for 30 min. 99m Tc]NaTcO4 physiological saline solution) into a freeze-dried kit, then add freshly washed [ 99m [Tc] NaTcO4 in physiological saline (approximately 185-3700 MBq, 0.5 mL) was heated in a boiling water bath for 30 minutes. The reaction solution was cooled and transferred to a SepPak C18 Plus Light cartridge (Waters Corporation, pretreated with 10 mL of anhydrous ethanol and 10 mL of water). The cartridge was then washed with water (5 mL) and eluted with 1 mL of a mixture of ethanol and water (0.75 / 0.25, v / v). The eluate was purified by the following HPLC separation method to obtain the radiopharmaceutical.

[0121] HPLC semi-preparative column separation and purification conditions: Phase A: water (containing 0.1% trifluoroacetic acid), Phase B: acetonitrile (containing 0.1% trifluoroacetic acid), gradient: 5% Phase B (0-4 min), 5% to 60% Phase B (4-26 min), and 60% Phase B (26-30 min). Total flow rate: 2.363 mL / min. Beijing Huideyi QuikSep SP ODS-A reversed-phase C18 semi-preparative column, 10 μm particle size, column size 10 mm x 250 mm.

[0122] The HPLC analytical column conditions for the two types of radiopharmaceuticals mentioned above were: phase A was water (containing 0.1% trifluoroacetic acid), phase B was acetonitrile (containing 0.1% trifluoroacetic acid), and the gradient was: 0-2 min phase B: 10%, 2-10 min phase B: 10% to 90%, 10-15 min phase B: 90%, 15-20 min phase B: 90% to 10%, and 20-30 min phase B: 10%. Total flow rate: 1 mL / min. Tianjin Bona Aijieer Venusil MP C18(2) reversed-phase C18 analytical column, column packing particle size 5 μm, 4.6 mm*250 mm.

[0123] like Figures 6-10 As shown, under the HPLC analysis conditions, the peak times of the above radioactive drugs were approximately 11.796 min, 11.925 min, 10.378 min, 10.526 min and 10.282 min, respectively.

[0124] Example 6: In vitro stability study of radiopharmaceuticals

[0125] 1) Stability test in physiological saline containing 10% ethanol:

[0126] Stability in normal saline (room temperature): Take out about 4 μCi of the above radioactive drug in normal saline at 1 hour and 2 hours respectively, and dilute it with 100 μL of a mixture of acetonitrile containing 1‰ trifluoroacetic acid and water containing 1‰ trifluoroacetic acid (1 / 19, v / v).

[0127] 2) Mouse serum stability test:

[0128] The marker (4 μCi) and saline (200 μL) solution were added to 400 μL of mouse serum and incubated at 37°C. After 2 h, the protein was precipitated with acetonitrile (200 μL) and the marker (150 μL) solution was analyzed by HPLC.

[0129] Radiopharmaceuticals 99m When [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPTS was placed in normal saline and mouse serum at room temperature for 6 h, the area ratios of its prototype peak were 99.110% and 99.021% respectively under the above HPLC analysis conditions, both greater than 95%, indicating that it is stable in this system ( Figure 11 );

[0130] Radiopharmaceuticals 99m When [Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI46-PEG2-tricine_TPPTS was placed in normal saline and mouse serum at room temperature for 6 h, the area ratios of its prototype peak were 99.456% and 99.159% respectively under the above HPLC analysis conditions, both greater than 95%, indicating that it is stable in this system ( Figure 12 );

[0131] Radiopharmaceuticals 99m When [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_TPPTS was placed in normal saline and mouse serum at room temperature for 6 h, the area ratios of its prototype peak were 99.372% and 99.257% respectively under the above HPLC analysis conditions, both greater than 95%, indicating that it is stable in this system ( Figure 13 );

[0132] Radiopharmaceuticals 99mWhen [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_EDDA was placed in normal saline and mouse serum at room temperature for 6 h, the area ratios of its prototype peak were 98.663% and 98.347% respectively under the above HPLC analysis conditions, both greater than 95%, indicating that it is stable in this system ( Figure 14 );

[0133] Radiopharmaceuticals 99m When [Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPTS was placed in normal saline and mouse serum at room temperature for 6 h, the area ratios of its prototype peak were 98.975% and 98.768% respectively under the above HPLC analysis conditions, both greater than 95%, indicating that it is stable in this system ( Figure 15 ).

[0134] Example 7: Biodistribution experiment

[0135] Biodistribution studies were performed using female BALB / c nude mice bearing U87 MG tumors. BALB / c nude mice were purchased from Beijing Weitong Lihua Co., Ltd., China. Approximately 5×10 6 U87 MG cells were subcutaneously injected into the left anterior axilla of female mice. Approximately 3 weeks later, when the tumor volume reached 100 mm 3 Can be used for experiments.

[0136] 1. Preparation of injection solution: Inject 7.4 MBq / 160 μL per mouse.

[0137] 2.12 U87MG tumor-bearing nude mice (BALB / c mice) were divided into 4 experimental groups, with an average of 3 female mice in each group.

[0138] 3. Time point for organ and tissue sampling: The mice in the four groups were killed 60 minutes after injection, and tissue samples were collected.

[0139] 4. The radioactivity count was measured by γ-counter and the data was processed. The results are shown in Table 1 below.

[0140] Where A is [ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-tricine_TPPTS B is [ 99m Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI46-PEG2-tricine_TPPPTS

[0141] C is [ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_TPPTS D is [ 99m Tc]Tc-HYNIC-VI-3-OSu-VI-8-de_Fmoc-FAPI46-PEG6-tricine_EDDA E is [ 99m Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI04-PEG6-tricine_TPPPTS

[0142] Table 1 Biodistribution data of radiopharmaceuticals in female U87MG tumor-bearing BALB / c mice 60 min after tail vein injection (mean ± SD, n = 3)

[0143]

[0144]

[0145] [a] Each mouse was injected with 7.4 MBq by tail vein. Except for the gastrointestinal uptake value of %ID, the other tissue absorption values ​​were %ID / g. 99m Tc]Tc-HYNIC-VI-3-OSu-D-Pro-FAPI46-PEG2-tricine_TPPTS) showed relatively good efficacy 1 h after tail vein injection, with a high tumor uptake value (17.71±3.09% ID / g). At this time, the tumor / flesh, tumor / liver, and tumor / bone values ​​were also relatively ideal, at 6.09±0.26, 4.33±0.38, and 4.17±1.20, respectively.

[0146] In addition, the tumor / meat and tumor / bone values ​​of compound A were also ideal, at 7.01±0.22 and 4.10±0.57, respectively; the tumor / bone value of compound D was also relatively high, at 4.30±1.44.

[0147] In the present invention, by simultaneously introducing structures such as X into the general formula, the structure of the present application maintains good stability, high tumor uptake, and a high on-target / off-target ratio, making it suitable for diagnosing or treating diseases characterized by FAP overexpression. More importantly, the radiopharmaceutical of the present application has the advantages of one-step labeling, no need for drying and dehydration, and a simple purification process.

[0148] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent replacements or improvements made within the spirit and principles of the present application should be included in the scope of protection of the present application.

Claims

1. A FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group, characterized in that: comprising FAPI04 or FAPI46, and a 6-hydrazinonicotinamide group connected via a linker; Preferably, the linker is glycine-L-allylglycine, glycine-L-allylglycine-polyethylene glycol or proline-polyethylene glycol.

2. A 6-hydrazinonicotinamide-containing FAPI04 or FAPI46 derivative according to claim 1, characterized in that: The general structural formula of the FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group is shown in the following formula (I): Among them, when R 1 When it is O, X is selected from When R 1 for When X is selected from n is any integer from 1 to 6.

3. Use of a 6-hydrazinonicotinamide-containing FAPI04 or FAPI46 derivative according to claim 1 or 2 in the preparation of a drug for treating cancer.

4. A FAPI04 or FAPI46 derivative labeled with radionuclide M, characterized in that: It is formed by coordinating the FAPI04 or FAPI46 derivative containing a 6-hydrazinonicotinamide group as claimed in claim 1 or 2, a co-ligand and a radionuclide M.

5. A radionuclide M-labeled FAPI04 or FAPI46 derivative according to claim 4, characterized in that: The co-ligand is tris(hydroxymethyl)glycine and sodium triphenylphosphine trisulfonate, or tricine and ethylenediamine-N,N'-diacetic acid.

6. A radionuclide M-labeled FAPI04 or FAPI46 derivative according to claim 4, characterized in that: The radionuclide M is a metal radionuclide; Preferably, the radionuclide M is 99m Tc, 99 Tc, 94m Tc, 94 Tc, 52 Mn, 186 Re or 188 Re; More preferably, the radionuclide M is 99m Tc.

7. The radionuclide M-labeled FAPI04 or FAPI46 derivative according to claim 4, characterized in that: The structural formula of the FAPI04 or FAPI46 derivative labeled with the radionuclide M is shown in the following formula (II) or (III): Among them, when R 1 When it is O, X is selected from When R 1 for When X is selected from n is any integer from 1 to 6.

8. A tumor imaging agent or anti-tumor radiopharmaceutical, characterized in that: Comprising a FAPI04 or FAPI46 derivative labeled with the radionuclide M as described in any one of claims 4 to 7.

9. Use of the FAPI04 or FAPI46 derivative labeled with the radionuclide M according to any one of claims 4 to 7 in the preparation of a tumor imaging agent.

10. Use of the FAPI04 or FAPI46 derivative labeled with the radionuclide M according to any one of claims 4 to 7 in the preparation of anti-tumor drugs.

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