FAP-targeted aluminum fluoride labeled compound and application thereof

Abstract

The invention relates to a FAP-targeted aluminum fluoride labeled compound and application thereof, the FAP-targeted aluminum fluoride labeled compound has a structure as shown in a general formula (I) or a pharmaceutically acceptable salt thereof, X is selected from O or # imgabs0 #, and R11 is selected from H, alkyl and halogenated alkyl; r1 is selected from amido, carbonyl, amino, carboxyl, peroxy, disulfide, C2-C15 alkyl containing or not containing substituent groups, C3-C15 heteroalkyl, C3-C15 cycloalkyl, C4-C15 heterocycloalkyl, C6-C15 aryl or C5-C15 heteroaryl. The formula (I) is shown in the description. R2 is a group formed by complexing a residue of the ligand with nuclides, the nuclides are preferably 18F and 19F, and the nuclides are complexed after forming multiple radical ions AlF < 2 + >. The present invention is useful in the diagnosis or treatment of diseases characterized by the overexpression of fibroblast activating protein (FAP). # imgabs1 #

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and in particular to an aluminum fluoride labeled compound targeting FAP and application thereof. Background Art

[0002] Fibroblast activation protein (FAP) is highly expressed in the vast majority of 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, which have attracted extensive attention and research.

[0003] 68 Ga-FAPI has been proposed as a promising PET tracer. However, the use 68 Ga-labeled radiotracers for PET imaging have disadvantages in terms of radionuclide supply. 68 Ga 3+ Produced by a generator, the yield is low and the half-life is short (only 68 minutes), which makes it impossible to achieve mass production and distribution, greatly limiting its diagnostic application in clinical promotion. The connecting chain between the targeting group and the chelating agent plays an important role in regulating the biological properties of radioactive drugs in the body. Radioactive drugs targeting FAP will be of great significance for the early diagnosis of tumors such as colorectal cancer, pancreatic cancer, ovarian cancer and melanoma. Summary of the invention

[0004] The connecting chain between the targeting group and the chelating agent plays an important role in regulating the biological properties of radiopharmaceuticals in the body. The present invention uses various groups such as aliphatic carbon chains, ethylene glycol chains, and six-membered ring carbon chains as connecting agents to connect the targeting group and the chelating agent, and designs and synthesizes multiple radiolabeled derivatives to explore their potential in tumor imaging.

[0005] One of the inventive points of the present invention is: an aluminum fluoride labeled compound targeting FAP, having a structure shown in general formula (I) or a pharmaceutically acceptable salt thereof,

[0006]

[0007] Wherein, X is selected from O or Among them, R 11 is selected from H, alkyl, and haloalkyl;

[0008] R1 is selected from -CO-NH-, -NH-CO-, -CO-, -NH-, -C(O)O-, -OC(O)-, -OO-, -SS-, C2 to C3 with or without substituents 15 Alkyl, C3~C 15 Heteroalkyl, C3~C 15 Cycloalkyl, C4~C 15 Heterocycloalkyl, C6~C 15 Aryl or C5~C 15 Heteroaryl, C3~C 15 Cycloalkyl-C2~C 15 Alkyl, C4~C 15 Heterocycloalkyl-C2~C 15 Alkyl, C6~C 15 Aryl-C2~C 15 Alkyl, C5~C 15 Heteroaryl-C2~C 15 alkyl..

[0009] The heteroatom contained in the heteroalkyl group, heterocycloalkyl group or heteroaryl group is selected from an oxygen atom, a nitrogen atom, a phosphorus atom or a sulfur atom; preferably, the heteroatom is an oxygen atom or a nitrogen atom.

[0010] The substituent is halogen, amide, amino, hydroxyl, carboxyl, nitro, cyano, hydrocarbon, alkoxy, alkylthio, C3-C 10 Cycloalkyl, C4~C 10 Heterocycloalkyl, C6~C 10 Aryl or C5~C 10 heteroaryl;

[0011] R2 is a group formed by the complexation of the residue of the ligand with the nuclide; wherein the nuclide is preferably 18 F. 19 F, nuclide is the complex ion AlF 2+ Re-complexed.

[0012] Complex ions are molecules or ions with different properties from the original components formed by a certain number of ligands bound to the central ion through coordination bonds. Nuclides directly serve as complex ions, such as Fe 2+ Mg 2+ Metal ions, or complex ions, such as AlF 2+ .

[0013] 19 F is a cold standard compound, generally 18 F will be tested before radioactive labeling 19 F, perform mass spectrometry confirmation to determine whether the compound can be labeled.

[0014] Further, X is selected from O or Among them, R 11 Selected from H, methyl, ethyl, propyl;

[0015] R1 is selected from

[0016] Among them, Y is selected from R3 is selected from methyl, ethyl, propyl;

[0017] Z is selected from -CO-NH-, -NH-CO-, -CO-, and R4 is selected from

[0018] R5 is selected from

[0019] q is an integer of 1-10; n is an integer of 1-10.

[0020] R2 is a group formed by the complexation of a nuclide with a residue of a NOTA ligand, a DOTA ligand, a DTPA ligand, a HYNIC ligand or a DFO ligand, wherein the nuclide is Al 18 F or Al 19 F.

[0021] NOTA ligand has a CAS number of 56491-86-2. NOTA, the full name of which is N-hydroxysuccinimide ester, is a compound with multiple functional groups that give it good reactivity and selectivity. It is often used as a ligand to separate and purify metal ions by forming a stable complex with metal ions.

[0022] DOTA ligand is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetracarboxylic acid (DOTA for short), a twelve-membered tetraaza macrocyclic ligand, which is essentially a product in which the hydrogens on the four nitrogens of cycloamine are replaced by acetic acid groups. This molecule was first reported in 1976. When it was discovered, it was proven that it could form chelates with Ca2+ and Gd3+ with the largest known stability constants. The improved derivatives of this ligand can be used as bifunctional chelating agents.

[0023] DTPA (diethylenetriaminepentaacetic acid) ligand is an important multidentate ligand with 3 N atoms and 5 -COOH structures in its molecular structure. It is a commonly used bifunctional integrator in nuclear medicine. The anhydride form of DTPA, diethylenetriaminepentaacetic acid dianhydride (DTPAA), contains groups that easily react with polysaccharide hydroxyl groups.

[0024] HYNIC ligand is a ligand used for tumor molecular imaging. It consists of two components: Hynic (6-hydrazinobutyramide) and PSMA (prostate-specific membrane antigen). HYNIC is a compound used to attach radioisotopes to target molecules, such as 188Re-HYNIC-PSMA. PSMA is a membrane antigen that is specifically expressed on the surface of prostate cancer cells. Therefore, HYNIC-PSMA can be used for prostate cancer research and tumor molecular imaging.

[0025] DFO ligand, also known as deferoxamine, is an important iron chelator commonly used in the biomedical field. It is a hexadentate macrocyclic ligand with hydrophilic properties that can form a stable complex with iron ions, thereby effectively chelating and removing iron ions from the body. DFO ligands are widely used, including in metal coordination chemistry, radioisotope labeling, and biomedical research.

[0026] Further, X is selected from O or or N-CH3;

[0027] R1 is selected from

[0028] R2 is a group formed by the complexing nuclide with the residue of NOTA ligand.

[0029] The residue is the ligand. Since its carboxyl group participates in the formation of the amide bond, the part of the ligand minus the reacted carboxyl group is called the "residue". For example, the residue is

[0030] Furthermore, the structure in the general formula (I) Selected from R1 is selected from

[0031] Preferably, if the R2 structure comprises Al 19 F, the compound is a non-radioactive compound.

[0032] Furthermore, the structure Includes the following groups:

[0033]

[0034]

[0035] The second invention of the present invention is a precursor compound of an aluminum fluoride labeling compound targeting FAP, having a structure shown in the general formula (I), wherein X is consistent with X defined in any one of claims 1 to 3; R1 is consistent with R1 defined in any one of claims 1 to 3, and R2 is a structure containing an 8-10 membered heterocyclic ring, wherein the heterocyclic ring contains at least one N, preferably containing The structure of , wherein R7, R8, and R9 are independently H, carboxyl, amide, amino, hydroxyl, nitro, or optionally substituted alkyl, alkoxy, or alkylthio.

[0036] Preferably, R2 is selected from

[0037] Specifically, the structure The precursor compound of the aluminum fluoride labeled compound includes a connecting chain and a chelating agent, and the specific structure is shown below:

[0038]

[0039] Furthermore, the precursor compound includes any one of the following compounds:

[0040]

[0041] The third invention of the present invention is: a PET imaging agent, comprising the above-mentioned aluminum fluoride labeled compound targeting FAP, preferably Al 18 F.Al 19 F-labeled compounds.

[0042] The fourth inventive point of the present invention is the use of the above-mentioned aluminum fluoride-labeled compound targeting FAP in the preparation of drugs for radioactive diagnosis and treatment of diseases.

[0043] The fifth invention point of the present invention is: use of the above-mentioned aluminum fluoride labeled compound targeting FAP in the preparation of tumor diagnostic imaging agents.

[0044] The present invention uses various groups such as aliphatic carbon chains, ethylene glycol chains, and six-membered ring carbon chains as linkers to connect the targeting group and the chelating agent, and designs and synthesizes multiple radiolabeled aluminum fluoride derivatives to explore their potential in tumor imaging.

[0045] Compared with the prior art, the present invention has the following advantages:

[0046] The aluminum fluoride-labeled compound targeting FAP of the present invention is used for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein (FAP). 18 The positron range of F is shorter than 68Ga can both reduce image noise and improve contrast resolution, thereby achieving higher spatial resolution of PET / CT imaging and maximizing the possibility of detecting subtle lesions. 18 F]AlF radiolabeling strategy is a new type of positron nuclide labeling technology based on [ 18 F][AlF] 2+ The formation of cations and complexation with the bifunctional chelator NOTA has the advantages of one-step labeling, no need for drying and water removal, and simple purification process. The compounds of the present application can maintain good in vitro stability, and the tumor uptake is significantly improved; the uptake of the present invention in tumors is specific, and the binding of radioactive drugs to FAP-α receptors can be competitively replaced by active drug precursors at the same target, indicating that it can be used as a new type of PET molecular probe with excellent pro-tumor performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 [ 18 F]-HPLC profile of in vitro stability of compound 2;

[0048] Figure 2 [ 18 F]-in vitro stability HPLC profile of compound 3;

[0049] Figure 3 [ 18 F]-HPLC profile of in vitro stability of compound 4;

[0050] Figure 4 (A) U87MG mice were injected with 18 F]-Coronal PET images 60 min after compound 3; (B) U87MG mice were injected with [ 18 (B) Coronal PET images 60 min after [F]-compound 3 and NOTA-FAPI-42.

[0051] Specific implementation method

[0052] In order to make the purpose, technical solution and advantages of the present application more clear, the present application is further described in detail below. However, it should be understood that the description here is only used to explain the present application and is not used to limit the scope of the present application.

[0053] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present application, and the terms used herein in the specification of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The reagents and instruments used herein are all commercially available, and the characterization means involved can refer to the relevant descriptions in the prior art, which will not be repeated herein.

[0054] In order to further understand the present application, the present application is further described in detail below in conjunction with the best embodiment.

[0055] Example 1 Synthesis of Compound 1

[0056]

[0057] 1) Synthesis of Compound II-3a

[0058] Compound II-2a (317 mg, 1.23 mmol, 1.5 equiv) was dissolved in DMF (10 mL) solution, HATU (468 mg, 1.23 mmol, 1.5 equiv) and DIPEA (106 mg, 1.64 mmol, 2 equiv) were added, and the mixture was stirred at room temperature for 5 minutes, and compound II-1 (400 mg, 0.82 mmol, 1 equiv) was added. The mixture was reacted at room temperature overnight, and the mixture was dried by oil pump at 50°C. The mixture was separated by silica gel column chromatography (dichloromethane: methanol = 20:1 to 10:1) to give compound II-3a (400 mg, white solid, yield 67.21%). 1 H NMR (400MHz, Methanol-d4) δ8.75(d,J=4.4Hz,1H),7.97(d,J=9.3Hz,1H),7.94(d,J=2.6Hz,1H),7.5 6(d,J=4.4Hz,1H),7.46(dd,J=9.2,2.7Hz,1H),5.12(dd,J=9.3,3.0Hz,1H),4.34–4.11(m,6H),3.60( s,4H),2.95–2.79(m,4H),2.63(q,J=8.6,8.0Hz,3H),2.54(s,2H),2.48(t,J=4.7Hz,2H),2.08(d,J= 6.8Hz,1H),1.79(t,J=14.4Hz,4H),1.48(d,J=13.0Hz,4H),1.43(s,9H),1.07–0.97(m,2H).HRMS(ESI + ):m / z calculated for[C 37 H 49 F2N7O6+H] + 726.3785, found726.3778.

[0059] 2) Synthesis of Compound II-4a

[0060] Trifluoroacetic acid (1 mL) was added to a solution of compound II-3a (297 mg, 0.41 mmol) in dichloromethane (1 mL), and the mixture was stirred at 30°C for 2 h. After TLC detection showed that there was no more raw material, the solvent was removed by spin drying, and the mixture was dissolved in a small amount of dichloromethane and concentrated under reduced pressure to remove the solvent. This was repeated twice to obtain compound II-4a (yellow oil, yield 99%), which was used directly in the next step without purification. HRMS (ESI + ):m / z calculated for[C 32 H 41 F2N7O4+H] + 626.326; found 626.3259.

[0061] 3) Synthesis of Compound II-5a

[0062] Compound 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)acetic acid (120 mg, 0.29 mmol, 1 equiv) was dissolved in DMF (5 mL) solution, HATU (132 mg, 0.348 mmol, 1.2 equiv) and DIPEA (187 mg, 1.45 mmol, 5 equiv) were added, and the mixture was stirred at room temperature for 5 minutes, and then a DMF solution (1 mL) of compound II-4a (181 mg, 0.29 mmol, 1 equiv) was added, and the mixture was stirred at room temperature for 6 hours. The reaction progress was monitored by TLC, and water was added to terminate the reaction and the mixture was extracted with ethyl acetate (10 mL*3 times). The organic layer was collected and dehydrated, and silica gel was added and dried under reduced pressure, and finally separated by column chromatography (dichloromethane: methanol = 20:1 to 10:1) to obtain compound II-5a (90 mg, light yellow solid, yield 30.33%). 1 HNMR(400MHz, Methanol-d4)δ8.75(d,J=4.4Hz,1H),7.97(d,J=9.3Hz,1H),7.95(d,J=2.6Hz,1H),7. 57(d,J=4.5Hz,1H),7.48–7.44(m,1H),5.16–5.10(m,1H),4.32–4.14(m,6H),3.61(s,4H),3.52–3.33 (m,6H),3.11(d,J=6.8Hz,2H),2.98–2.76(m,12H),2.67–2.62(m,3H),2.55(s,2H),2.51–2.47(m,2H) ,2.12–2.07(m,2H),1.88–1.73(m,5H),1.48(s,18H),1.34–1.26(m,4H),1.12–1.05(m,2H).HRMS(ESI+ ):m / z calculated for[C 52 H 76 F2N 10 O9+H] + 1023.5837, found 1023.5843.

[0063] 4) Synthesis of Compound 1

[0064] Compound II-5a (31 mg, 0.03 mmol) was added to a 25 ml round-bottom flask, dichloromethane (1 mL) was added as solvent, and then an equal volume of trifluoroacetic acid was added. The reaction was stirred at 30°C for 2 h. After TLC detection showed that there was no more starting material, the solvent was removed by concentration under reduced pressure. The product was separated and purified using a C18 reverse phase separation column (water: methanol = 10:1 to 2:1) to obtain product 1 (12 mg, light yellow solid, yield 43.91%). 1 H NMR(400MHz,D2O)δ8.72(d,J=4.2Hz,1H),7.93(d,J=9.3Hz,1H),7.58(d,J=4.3Hz,1H),7.53(s,1H), 7.44(d,J=9.3Hz,1H),5.15(d,J=4.7Hz,1H),4.32(d,J=5.9Hz,2H),4.28–4.05(m,4H),3.99–3.65(m, 4H),3.56(s,4H),3.41(s,2H),3.39–3.32(m,4H),3.29(s,2H),3.20–2.92(m,12H),2.81–2.64(m,5H) ,2.28(s,2H),1.78(s,2H),1.76(s,2H),1.55(s,1H),1.46–1.32(m,2H),1.10–0.95(m,2H).HRMS(ESI - ):m / zcalculated for[C 44 H 60 F2N 10 O9+H] + 911.4585, found 911.4578.

[0065] Example 2 Synthesis of Compound 2

[0066] 1) Synthesis of compound II-3b

[0067] The synthesis method of II-3b is the same as that of compound II-3a, using compound II-1 and II-2b as raw materials, and the reaction progress is detected by TLC. The reaction solution is added into water to terminate the reaction, and then extracted with ethyl acetate (10mL*3 times), and washed with saturated brine. The organic phase is collected, added with silica gel, and then spin-dried, and separated by column chromatography (dichloromethane: methanol = 20: 1 to 10: 1) to obtain the product II-3b (403 mg, light yellow solid, yield 70.23%). 1 H NMR (400MHz, Methanol-d4) δ8.82(d,J=4.5Hz,1H),8.06(d,J=2.3Hz,1H),8.02(d,J=9.3Hz,1H),7.64(d, J=4.5Hz,1H),7.54–7.50(m,1H),5.17–5.10(m,1H),4.39(t,J=5.5Hz,2H),4.33(d,J=3.7Hz,2H),4.26–4 .10(m,2H),4.07–3.70(m,2H),3.63–3.32(m,6H),3.03(t,J=6.9Hz,2H),2.99–2.81(m,4H),2.44(t,J=7. 4Hz,2H),2.40–2.34(m,2H),1.65–1.58(m,2H),1.52–1.47(m,2H),1.43(s,9H),1.39–1.34(m,2H).MS(ESI + ):m / z calculated for[C 35 H 47 F2N7O6+H] + 700.3628; found 700.3635.

[0068] 2) Synthesis of compound II-4b

[0069] The synthesis method of II-4b is the same as that of compound II-4a, using compound II-3b as raw material to obtain the product II-4b (yellow oil, yield 99%).

[0070] 3) Synthesis of Compound II-5b

[0071] The synthesis method of II-5b is the same as that of compound II-5a, using compound II-4b and 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)acetic acid as raw materials, and the product is separated and purified by silica gel column chromatography (dichloromethane: methanol = 20:1 to 10:1) to obtain II-5b (121 mg, light yellow solid, yield 41.84%). 1H NMR (400MHz, Methanol-d4) δ8.74(d,J=4.3Hz,1H),7.96(d,J=9.3Hz,1H),7.95–7.92(m,1H),7.56(d,J=4.3Hz,1H),7. 47–7.43(m,1H),5.13(d,J=6.8Hz,1H),4.31(d,J=4.5Hz,2H),4.27(d,J=6.2Hz,2H),4.25–4.08(m,2H),3.58(d,J=12.1 Hz,4H),3.36(d,J=10.3Hz,4H),3.23(t,J=6.9Hz,4H),2.93–2.72(m,12H),2.66–2.60(m,4H),2.53(s,2H),2.48(s,2H ),2.39(t,J=7.3Hz,2H),2.08(s,2H),1.63–1.59(m,2H),1.56–1.53(m,2H),1.47(s,18H),1.40–1.35(m,2H).HRMS(ESI + ):m / z calculated for[C 50 H 74 F2N 10 O9+H] + 997.5681, found 997.5689.

[0072] 4) Synthesis of Compound 2

[0073] The synthesis method of compound 2 is the same as that of compound 1, using compound II-5b as the raw material, and the product is separated and purified by C18 reverse phase separation column (water: methanol = 10: 1 to 2: 1) to obtain compound 2 (23 mg, light yellow solid, yield 86.63%). 1HNMR(600MHz,D2O)δ8.70(d,J=4.1Hz,1H),7.88(d,J=9.1Hz,1H),7.56(d,J=4.1Hz,1H),7.48(s,1H),7.39(d,J=9.2Hz ,1H),5.21–5.11(m,1H),4.31(d,J=11.5Hz,2H),4.27(d,J=4.8Hz,1H),4.19(s,2H),4.17–4.05(m,1H),3.84(d,J=40. 6Hz,4H),3.54(s,4H),3.37(s,6H),3.31(s,2H),3.19(t,J=6.9Hz,4H),3.04(s,4H),3.01–2.92(m,4H),2.74(s,2H),2 .66(s,2H),2.46(t,J=7.2Hz,2H),2.28(s,2H),1.61–1.57(m,2H),1.54–1.50(m,2H),1.33(d,J=6.7Hz,2H).HRMS(ESI + ):m / z calculated for[C 42 H 58 F2N 10 O9+H] + 885.4429, found 885.4414.

[0074] Example 3 Synthesis of Compound 3

[0075] 1) Synthesis of compound II-3c

[0076] The synthesis method of II-3c is the same as that of compound II-3a, using compound II-1 and compound II-2c as raw materials, and the product is obtained by silica gel column chromatography separation (dichloromethane: methanol = 20:1 to 5:1) to obtain II-3c (516 mg, white solid, yield 85.99%). 1 H NMR (400MHz, Methanol-d4) δ8.75(s,1H),7.97(d,J=13.3Hz,2H),7.57(s,1H),7.46(d,J=8.2Hz,1H),5.13(d,J=6.9Hz,1H),4.30(s,4H),4.26(s ,2H),3.76–3.58(m,8H),3.55–3.48(m,4H),3.22(s,2H),2.96–2.72(m, 2H),2.66(s,2H),2.59–2.51(m,4H),2.10(s,2H),1.43(s,9H).HRMS(ESI +):m / z calculated for[C 35 H 47 F2N7O8+H + 732.3526; found 732.3532.

[0077] 2) Synthesis of compound II-4c

[0078] The synthesis method of II-4c is the same as that of compound II-4a, using compound II-3c as the raw material to obtain the product II-4c (yellow oil, yield 99%). MS (ESI + ):m / z calculated for[C 30 H 39 F2N7O6+H] + 632.30, found 632.38.

[0079] 3) Synthesis of compound II-5c

[0080] The synthesis method of II-5c is the same as that of compound II-5a, using compound II-4c and 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)acetic acid as raw materials, and the product is obtained by silica gel column chromatography (dichloromethane: methanol = 20:1 to 5:1) to obtain II-5c (130 mg, light yellow solid, yield 43.56%). 1 H NMR (600MHz, Methanol-d4) δ8.74(d,J=3.4Hz,1H),7.96(d,J=9.1Hz,1H),7.93(s,1H),7.56(d,J=3.2Hz,1H ),7.45(d,J=9.2Hz,1H),5.12(d,J=9.2Hz,1H),4.31(s,2H),4.28(t,J=5.1Hz,2H),4.25(s,2H),4.22(d,J=4 .0Hz,1H),4.17–4.09(m,1H),3.69–3.62(m,8H),3.60(s,2H),3.57–3.55(m,2H),3.50(s,2H),3.40(s,2H), 3.16–2.75(m,16H),2.66–2.63(m,2H),2.55(s,2H),2.51(s,2H),2.10–2.07(m,2H),1.48(s,18H).HRMS(ESI + ):m / z calculated for[C 50 H 74 F2N 10 O 11 +H]+ 1029.5579, found 1029.5584.

[0081] 4) Synthesis of Compound 3

[0082] The synthesis method of compound 3 is the same as that of compound 1, using compound II-5c as the raw material, and the product is separated and purified by C18 reverse phase separation column (water: methanol = 10: 1 to 1: 1) to obtain compound 3 (23 mg, light yellow solid, yield 83.61%). 1 HNMR(600MHz,D2O)δ8.60(d,J=4.4Hz,1H),7.80(d,J=9.2Hz,1H),7.46(d,J=4.6Hz,1H),7.40(d,J=2.6 Hz,1H),7.32–7.29(m,1H),5.05–5.01(m,1H),4.22(s,2H),4.20(d,J=11.0Hz,2H),4.18–4.14(m,1H),4 .12–4.09(m,2H),4.06–3.99(m,1H),3.66(s,2H),3.57(s,4H),3.48(t,J=5.9Hz,2H),3.40(s,4H),3.3 0–3.17(m,12H),3.00(s,2H),2.89–2.79(m,8H),2.59(s,2H),2.53(s,2H),2.19–2.15(m,2H).HRMS(ESI + ):m / z calculated for[C 42 H 58 F2N 10 O 11 +H] + 917.4327, found 917.4335.

[0083] Example 4 Synthesis of Compound 4

[0084]

[0085] 1) Synthesis of Compound II-9a

[0086] Cool the DMF (500 mL) solution of compound II-7a (58 g, 141 mmol, 1 equiv) to 0 °C under an ice bath, add HATU (64 g, 169.2 mmol, 1.2 equiv) and stir for 5 minutes, then slowly add DIPEA (55 g, 423 mmol, 3 equiv) and compound II-8 (23 g, 141 mmol, 1 equiv), gradually warm to room temperature for 4 hours, and monitor the reaction progress by TLC. For post-treatment, add the DMF solution to water (1.5 L), then extract with ethyl acetate (100 mL * 3 times), and wash the organic layer with saturated brine. Collect the organic phase, dry and remove water, add silica gel and decompress and spin dry, and finally separate and purify by column chromatography (petroleum ether: ethyl acetate = 5: 1 to 3: 1) to obtain compound II-9a (63 g, light yellow solid, yield 79.98%). 1 H NMR (400MHz, CDCl3) δ7.77(s,1H),7.76(s,1H),7.60(s,1H),7.58(s,1H),7.40–7.29(m,9H),7.02(s,1H),5.98(d,J=8.1Hz,1H),5.17(s,2 H),4.58(s,1H),4.48–4.38(m,2H),4.23(t,J=6.9Hz,1H),4.13–4.02(m,2H),3.00–2.86(m,1H),2.73–2.57(m,1H),1.45(s,9H).HRMS(ESI + ):m / zcalculated for[C 32 H 34 N2O7+H] + 559.2438, found 559.2434.

[0087] 2) Synthesis of Compound II-10a

[0088] Morpholine (10.454 g, 120 mmol, 3 equiv) was added to DMF (100 mL) of compound II-9a (22 g, 40 mmol, 1 equiv), and the mixture was reacted at room temperature for 1 h. After TLC detection showed that there was no more raw material, the reaction solution was poured into water (500 mL), and then extracted with ethyl acetate (100 mL*3 times), and washed with saturated brine. The organic layer was collected and dried to remove water, and the liquid was removed by adding silica gel under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate = 3:1 to 1:1) to obtain compound II-10a (10.77 g, light yellow solid, yield 80.04%). 1H NMR (400MHz, CDCl3) δ7.92 (s, 1H), 7.35 (s, 5H), 5.17 (s, 2H), 4.08 (t, J = 5.6Hz, 2H), 3.7 9–3.67(m,1H),2.89–2.79(m,1H),2.60–2.44(m,1H),1.91(s,2H),1.44(s,9H).MS(ESI + ):m / z calculated for[C 17 H 24 N2O5+Na] + 359.16, found 359.17.

[0089] 3) Synthesis of Compound II-12a

[0090] The synthesis method of II-12a is the same as that of II-9a, using compounds II-10a and II-11a as raw materials, and the product is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1 to ethyl acetate) to obtain II-12a (11.1 g, light yellow solid, yield 60.1%). 1 H NMR(400MHz, CDCl3)δ7.76(s,1H),7.74(s,1H),7.59(s,1H),7.58(s,1H), 7.38(t,J=7.5Hz,2H),7.32–7.27(m,7H),5.10(s,2H),4.88(q,J=6.3Hz,1 H),4.41–4.36(m,2H),4.21(t,J=7.0Hz,1H),4.07–3.95(m,2H),3.88(t,J =5.0Hz,2H),2.89–2.81(m,1H),2.68–2.60(m,1H),1.41(s,9H).HRMS(ESI + ):m / z calculated for[C 34 H 37 N3O8+H] + 616.2653, found 616.2640.

[0091] 4) Synthesis of Compound II-13a

[0092] Compound II-12a (9.24 g, 15 mmol, 1 equiv) was added to a three-necked flask, dissolved with a mixed solution of methanol and tetrahydrofuran (50 mL, v:v = 1:1), 10% palladium carbon (0.9 g, 0.1 equiv) was added, hydrogen was replaced with a glass three-way joint and the reaction was stirred at room temperature. TLC was used to monitor the progress of the reaction. After the reaction was completed, the palladium carbon was filtered off and the liquid was dried under reduced pressure to obtain II-13a (7.88 g, black solid, yield 99%). The product did not need to be purified and could be directly used in the next step. HRMS (ESI + ):m / zcalculated for[C 27 H 31 N3O8+Na] + 548.2003,found 548.2001.

[0093] 5) Synthesis of Compound II-14a

[0094] A solution of compound II-13a (1.08 g, 2.06 mmol, 2 equiv) in DMF (10 mL) was cooled to 0°C under an ice bath, HATU (587 mg, 1.545 mmol, 1.5 equiv) was added and stirred for 5 minutes, then DIPEA (266 mg, 2.06 mmol, 2 equiv) and compound II-1 (500 mg, 1.03 mmol, 1 equiv) were slowly added, and the temperature was slowly raised to room temperature for 5 hours. The progress of the reaction was monitored by TLC. During post-treatment, DMF was poured into water (100 mL) to terminate the reaction, followed by extraction with ethyl acetate (30 mL*3 times) and washing with saturated NaCl solution. The ethyl acetate layers were combined and dried to remove water, and silica gel was added and the liquid was removed by vacuum rotary evaporation. II-14a (443 mg, light yellow solid, yield 43.31%) was obtained by column chromatography separation (dichloromethane: methanol = 20:1 to 10:1). 1H NMR (400MHz, Methanol-d4) δ8.73(d,J=4.4Hz,1H),7.96(d,J=9.2Hz,1H),7.91(d,J=2.5Hz,1H),7.77(d,J=7.5Hz,2H),7.65(d,J=7.4Hz, 2H),7.56(d,J=4.4Hz,1H),7.45–7.42(m,1H),7.36(t,J=7.4Hz,2H),7.28(t,J=7.4Hz,2H),5.13–5.09(m,1H),4.36(d,J=7.2Hz,1H),4.34 –4.28(m,4H),4.24–4.12(m,5H),4.08(d,J=11.8Hz,1H),3.89(d,J=16.8Hz,1H),3.81(s,2H),3.47(s,2H),3.41(s,2H),2.98–2.82(m,2H ),2.76(d,J=16.7Hz,1H),2.66–2.59(m,1H),2.57–2.51(m,2H),2.44(s,2H),2.40–2.33(m,2H),2.04–1.98(m,2H),1.42(s,9H).HRMS(ESI + ):m / z calculated for[C 51 H 57 F2N9O 10 +H] + 994.4269, found 994.4261.

[0095] 6) Synthesis of Compound II-15a

[0096] To a solution of compound II-14a (443 mg, 0.446 mmol) in dichloromethane (4 mL) was added piperidine (0.8 mL), and the mixture was reacted at room temperature for 2 h. After TLC detection showed that there was no more starting material, silica gel was added and the mixture was dried under reduced pressure. The mixture was separated by column chromatography (dichloromethane: methanol: 28% aqueous ammonia = 20: 1: 0.1 to 4: 1: 0.1) to give II-15a (251 mg, yellow oil, 72.92%). 1H NMR (400MHz, Methanol-d4) δ8.75(d,J=4.4Hz,1H),7.97(d,J=9.3Hz,1H),7.95(d,J=2.7Hz, 1H),7.57(d,J=4.5Hz,1H),7.49–7.44(m,1H),5.15–5.11(m,1H),4.84–4.81(m,1H),4.32–4. 05(m,8H),3.63–3.58(m,2H),3.54–3.50(m,2H),3.35–3.32(m,2H),2.99–2.68(m,4H),2.67– 2.63(m,2H),2.59–2.54(m,2H),2.53–2.49(m,2H),2.13–2.05(m,2H),1.44(s,9H).HRMS(ESI + ):m / z calculated for[C 36 H 47 F2N9O8+H] + 772.3588, found 772.3576.

[0097] 7) Synthesis of Compound II-16a

[0098] The synthesis method of II-16a is the same as that of II-5a, using compound II-15a and 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)acetic acid as raw materials to obtain II-16a (68 mg, light yellow solid, yield 42.76%). 1 H NMR(400MHz, Methanol-d4)δ8.74(d,J=3.9Hz,1H),7.96(d,J=9.2Hz,1H),7.94–7.91(m,1H) ,7.57(d,J=3.8Hz,1H),7.46(d,J=9.0Hz,1H),5.12(d,J=8.2Hz,1H),4.33–4.23(m,5H),4.1 2–3.83(m,6H),3.62(d,J=17.3Hz,4H),3.50(d,J=19.5Hz,4H),3.01–2.81(m,18H),2.65(s, 2H),2.58(s,2H),2.52(s,2H),2.09(s,2H),1.48(s,9H),1.47(s,9H),1.44(s,9H).HRMS(ESI + ):m / z calculated for[C 56 H 82 F2N 12 O13 +H] + 1169.6165, found1169.6174.

[0099] 8) Synthesis of Compound 4

[0100] The synthesis method of compound 4 is the same as compound 1, using II-16a as raw material to obtain the product compound 4 (13 mg, light yellow solid, yield 56.69%). 1 H NMR(600MHz,D2O)δ8.79(d,J=4.6Hz,1H),7.98(d,J=9.2Hz,1H),7.69(d,J=4. 6Hz,1H),7.58(s,1H),7.51(d,J=8.6Hz,1H),5.19–5.14(m,1H),4.77–4.74(m ,1H),4.42–4.06(m,12H),4.05–3.80(m,4H),3.69(d,J=9.0Hz,2H),3.56–3.3 2(m,8H),3.19(s,2H),3.11(s,2H),3.08–2.65(m,12H),2.34(s,2H).HRMS(ESI + ):m / z calculatedfor[C 44 H 58 F2N 12 O 13 +H] + 1001.4287, found 1001.4270.

[0101] Example 5 Synthesis of Compound 5

[0102] 1) The synthesis method of II-9b is the same as II-9a, using compounds II-7b and II-8 as raw materials to obtain compound II-9b (54.6 g, white solid, yield 67.62%). 1 H NMR (400MHz, CDCl3) δ7.77(s,1H),7.75(s,1H),7.60(s,1H),7.59(s,1H),7. 41–7.29(m,9H),6.86(s,1H),5.79(d,J=8.2Hz,1H),5.18(s,2H),4.44–4.33( m,2H),4.28(d,J=6.7Hz,1H),4.21(t,J=6.8Hz,1H),4.16–4.02(m,2H),2.51– 2.32(m,2H),2.11(d,J=6.6Hz,1H),2.01–1.89(m,1H),1.46(s,9H).HRMS(ESI +):m / zcalculated for[C 33 H 36 N2O7+H] + 573.2595, found 573.2583.

[0103] 2) Synthesis of Compound II-10b

[0104] The synthesis method of II-10b is the same as II-10a to obtain compound II-10b (11.5 g, light yellow solid, yield 82.04%). 1 H NMR (400MHz, CDCl3) δ7.35 (t, J = 2.5Hz, 5H), 5.17 (s, 2H), 4.68 (s, 2H), 4.07 (t ,J=5.3Hz,1H),2.38–2.33(m,2H),2.10(d,J=5.7Hz,2H),1.43(s,9H).MS(ESI + ):m / zcalculated for[C 18 H 26 N2O5+H] + 351.19, found 351.00.

[0105] 3) Synthesis of Compound II-12b

[0106] The synthesis method of II-12b is the same as that of II-9a, using compounds II-10b and II-11b as raw materials, and the product is obtained by silica gel column chromatography (petroleum ether: ethyl acetate = 3:1 to ethyl acetate) to obtain II-12b (15.35 g, light yellow solid, yield 72.49%). 1 H NMR (400MHz, DMSO-d6) δ8.50(d,J=7.9Hz,1H),8.44(t,J=5.9Hz,1H),7.96–7.92(m,1H),7.89(d,J =7.5Hz,2H),7.81(d,J=10.1Hz,2H),7.70(d,J=7.5Hz,2H),7.45–7.39(m,4H),7.37–7.30(m,7H),5 .13(s,2H),4.55–4.46(m,1H),4.34(d,J=6.9Hz,2H),4.24(d,J=6.2Hz,2H),4.23–4.10(m,1H),3.9 9–3.85(m,2H),2.32(t,J=7.8Hz,2H),2.10–2.00(m,1H),1.96–1.86(m,1H),1.37(s,9H).HRMS(ESI +):m / z calculated for[C 41 H 43 N3O8+H] + 706.3122, found 706.3117.

[0107] 4) Synthesis of Compound II-13b

[0108] The synthesis method of II-13b is the same as II-13a, using II-12b as the raw material to obtain compound II-13b (9.24 g, black solid, yield 99%). HRMS (ESI + ):m / z calculated for[C 34 H 37 N3O8+Na] + 638.2472, found 638.2462.

[0109] 5) Synthesis of Compound II-14b

[0110] The synthesis method of II-14b is the same as that of II-14a, using compound II-13b and raw material II-1 to obtain II-14b (564 mg, white solid, yield 50.5%). 1 H NMR (400MHz, Methanol-d4) δ8.71–8.66(m,1H),7.93–7.90(m,1H),7.87(s,1H),7.79(s,1H),7.73(d,J=6.9Hz,3H),7.59(s,1H ),7.57(s,1H),7.53–7.48(m,1H),7.42–7.36(m,3H),7.32(t,J=7.1Hz,2H),7.23(t,J=6.9Hz,2H),5.07(d,J=9.3Hz,1H),4.66– 4.61(m,1H),4.34–4.30(m,2H),4.29(s,2H),4.26(s,2H),4.19–4.05(m,5H),3.98(s,2H),3.47(s,2H),3.37(s,2H),2.96–2.7 2(m,2H),2.53–2.47(m,2H),2.43–2.35(m,6H),2.25–2.18(m,1H),2.09–2.00(m,1H),1.98–1.92(m,2H),1.37(s,9H).HRMS(ESI + ):m / z calculated for[C 58 H 63 F2N9O 10 +H] +1084.4738, found 1084.4715.

[0111] 6) Synthesis of Compound II-15b

[0112] The synthesis method of II-15b is the same as II-15a, using II-14b as raw material to obtain II-15b (314 mg, light yellow oil, 70.06%). 1 HNMR (400MHz, Methanol-d4) δ8.75(d,J=4.5Hz,1H),7.97(d,J=9.3Hz,1H),7.94(d,J=2.6Hz,1H),7.88(s,1H),7.79(d,J =7.8Hz,1H),7.57(d,J=4.4Hz,1H),7.55(s,1H),7.49–7.46(m,1H),7.46–7.44(m,1H),5.15–5.10(m,1H),4.69–4.65(m,1 H),4.32–4.19(m,6H),4.10(s,2H),3.90(s,2H),3.62–3.58(m,2H),3.54–3.51(m,2H),2.95–2.79(m,2H),2.66–2.61(m,2 H),2.57–2.54(m,2H),2.53–2.50(m,2H),2.45–2.41(m,2H),2.32–2.14(m,2H),2.07–2.02(m,2H),1.43(s,9H).HRMS(ESI + ):m / z calculated for[C 43 H 53 F2N9O8+H] + 862.4057, found 862.4051.

[0113] 7) Synthesis of Compound II-16b

[0114] The synthesis method of II-16b is the same as that of II-5a, using compound II-15b and 2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazacyclononane-1-yl)acetic acid as raw materials to obtain II-16b (87 mg, light yellow solid, yield 50.78%). 1H NMR (400MHz, Methanol-d4) δ8.75(d,J=4.4Hz,1H),7.97(d,J=9.3Hz,1H),7.94(d,J=2.7Hz,1H),7.81(d,J=1.8Hz, 1H),7.57(d,J=4.4Hz,1H),7.51–7.43(m,4H),5.14–5.10(m,1H),4.69–4.65(m,1H),4.50(s,2H),4.33–4.30(m,2H) ,4.30–4.10(m,6H),4.09(s,2H),3.60(d,J=6.0Hz,2H),3.53–3.51(m,2H),3.07–2.66(m,20H),2.63(d,J=7.6Hz,2H ),2.57–2.54(m,2H),2.52–2.50(m,2H),2.44–2.40(m,2H),2.09–2.05(m,2H),1.44(s,18H),1.43(s,9H).HRMS(ESI + ):m / z calculated for[C 63 H 88 F2N 12 O 13 +H] + 1259.6634, found 1259.6595.

[0115] 8) Synthesis of Compound 5

[0116] The synthesis method of compound 5 is the same as compound 1, using II-16b as raw material to obtain the product compound 5 (16 mg, light yellow solid, yield 50.5%). 1H NMR(600MHz,D2O)δ8.70(d,J=4.3Hz,1H),7.90–7.88(m,1H),7.71–7.68(m,1H),7.66(s,1H),7.58(d,J=4.4Hz,1 H),7.51–7.49(m,1H),7.46–7.39(m,3H),5.15–5.12(m,1H),4.53–4.51(m,1H),4.42(s,2H),4.35–4.24(m,4H),4 .19–4.10(m,6H),3.89–3.61(m,4H),3.46(s,2H),3.40(s,2H),3.38(s,2H),3.35(s,2H),3.08–2.79(m,12H),2. 73(s,2H),2.67–2.60(m,2H),2.44(t,J=7.2Hz,2H),2.27(s,2H),2.22–2.18(m,1H),2.12–2.07(m,1H).HRMS(ESI + ):m / z calculated for[C 51 H 64 F2N 12 O 13 +H] + 1091.4756, found 1091.4755.

[0117] Example 6 Compounds inhibit FAP enzyme activity in vitro (IC 50 )experiment

[0118] Table 1 Buffer preparation

[0119]

[0120] Prepare buffer: The final buffer consists of 50mM HEPES (2-[4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, purchased from Invitrogen) pH 7.4 buffer, 150mM sodium chloride solution, 1mM EDTA (Ethylenediaminetetraacetic acid, ethylenediaminetetraacetic acid), 0.1mg / mL BSA (Bovine Serum Albumin, purchased from Sigma) and ddH2O (double distilled water). Dilute the FAP enzyme to 0.75nM with detection buffer and dilute the Z-Gly-Pro-AMC (Z-glycylproline aminomethylcoumarin) substrate to 125μM. The test compound was serially diluted from 0.1μM to 5.08pM. 50μL of compound and 40μl of FAP enzyme were added to 384 wells. Then 10μl of Z-Gly-Pro-AMC substrate solution was added to each well. Seal the plate with stickers. Centrifuge at 600 rpm for 1 min. Incubate at room temperature for 60 min. FAPI-42 and UAMC-1110 (purchased from MCE) test IC 50 The buffer preparation method is consistent with the compounds of the present application. These two compounds are equivalent to positive drugs. Envision5 was used to read the data, and after measuring the fluorescence intensity of the Z-Gly-Pro-AMC substrate (Ex / Em=380 / 465nm), GraphPad Prism was used to analyze the data, draw the dose-response curve and calculate the IC 50 The values ​​are shown in Table 2.

[0121] Table 2 FAP enzyme inhibition activity test results

[0122]

[0123] For example, the half-inhibitory concentration of compound 3 for FAP is 0.513 nM, which is at the same nanomolar level as FAPI-42 and UAMC-1110 (0.424 nM and 0.886 nM). This shows that compound 3 has a strong in vitro inhibitory effect on FAP, and other compounds 1, 2, 4, and 5 also have a strong in vitro inhibitory effect on FAP.

[0124] Example 7 Radiolabeling Experiment

[0125] 1)[ 18 F]-compound radiolabeling operation steps: 0.5 mg of precursor (compounds 1, 2, 3, 4, 5) was dissolved in 100 μL of deionized water, and then 300 μL of acetonitrile, 20 μL of AlCl3 solution (10 mM) and 10 μL of glacial acetic acid were added. 18 F]F -((80-100 μL) was added to the mixture. The mixture was heated at 110°C for 15 minutes and then cooled. After dilution with 5 mL of deionized water, the reaction solution was transferred to a SepPak C18 Plus Light column (Waters Corporation, pretreated with 10 mL of anhydrous ethanol and 10 mL of water), then washed with water (5 mL) and eluted with 1 mL of a mixture of ethanol / water (0.75 / 0.25, v / v), concentrated by rotary evaporation, and then diluted with physiological saline containing 7% ethanol. The radiochemical purity of the labeled substance was analyzed by high performance liquid chromatography (HPLC). Taking the labeling of compound 2 as an example, the labeled 18 F-Compound 2 is shown below, and other compounds are not listed one by one.

[0126]

[0127] 2)[ 19 F]- compound labeling method: aluminum chloride (0.46 mg) and potassium fluoride (0.28 mg) were dissolved in 100 μL of deionized water, 0.5 mg of the precursor was dissolved in 300 μL of acetonitrile and then added, and acetic acid (10 μL) was added. The mixture was heated at 110°C for 15 minutes and identified by mass spectrometry. Take the labeling of compound 2 as an example. MS (ESI+): m / z calculated for [C 42 H 56 AlF3N 10 O9+H] + 929.41, found 929.50.

[0128]

[0129] Example 8 Fat-water distribution coefficient test

[0130] Equal volumes of n-octanol and phosphate buffer solution (PBS, pH=7.4) were added to a centrifuge tube, vortexed for 3-5 minutes, and then centrifuged at 5000 rpm for 5 minutes, and left to stand overnight at room temperature for use.

[0131] Take PBS (1000 μL), n-octanol (900 μL) and purified labeling solution (100 μL, 370 KBq) from the above mixture and add them to another centrifuge tube, centrifuge at 5000 rpm for 5 min, and vortex for 5 min. Collect 100 μL of each phase in a counting tube and measure with a γ counter (n=4). The calculation method of the distribution coefficient is as follows: P=(radioactivity of n-octanol layer / radioactivity of PBS layer). Log P is the mean ± standard deviation of 4 experiments.

[0132] Measured 18 F]-compound 2, [ 18F]-compound 3, [ 18 The lipid-water partition coefficients of [F]-compound 4 were -2.38±0.25, -2.70±0.06 and -3.28±0.04, respectively (as shown in Table 3).

[0133] Table 3 LogP test results of markers

[0134]

[0135] Example 9 Stability Study

[0136] 1) Stability test in physiological saline containing 10% ethanol.

[0137] The marker (1850 KBq) solution was placed in physiological saline (200 μL) and incubated at 37° C. After 1 and 2 h, the marker (50 μL) solution was directly analyzed by HPLC under the above HPLC conditions.

[0138] 2) Stability test of mouse serum.

[0139] The marker (1850 KBq) and saline (200 μL) solution were added to 400 μL mouse serum and incubated at 37° C. At 1 and 2 h, the protein was precipitated with acetonitrile (200 μL) and the marker (150 μL) solution was analyzed by HPLC.

[0140] The results of high performance liquid chromatography (HPLC) analysis showed that 18 In vitro stability HPLC analysis of compound 2 Figure 1 As shown, 18 In vitro stability HPLC analysis of compound 3 Figure 2 As shown, 18 In vitro stability HPLC analysis of compound 4 Figure 3 shown.

[0141] [ 18 The radioactive peak time of compound 2 was 23.35 min (0-15 min, 5% water (0.1% TFA) + 95% acetonitrile (0.1% TFA); 15-30 min, 95% water (0.1% TFA) + 5% acetonitrile (0.1% TFA), flow rate: 1 mL / min, SP ODS-A, 5 μm, 10×250mm). In the in vitro stability study at 37°C, the radiochemical purity of the marker after incubation in saline for 1h and 2h was 99.96% and 99.95%, respectively, and the radiochemical purity after incubation in mouse serum for 1h and 2h was 99.94% and 99.89%, respectively.

[0142] [ 18 The elution time of [F]-compound 3 was 22.63 min, and it had an excellent radiochemical purity of 99.99%. 18 The radiochemical purity of [F]-compound 3 in saline for 1 and 2 h was 99.98% and 99.97%, respectively, and the radiochemical purity of [F]-compound 3 in 37C for 1 and 2 h was 98.38% and 98.33%, respectively.

[0143] [ 18 The elution time of [F]-compound 4 was 23.71 min, and the radiochemical purity was 99.99%. 18 The in vitro stability test of compound 4 showed that after incubation in saline and mouse serum for 1 h and 2 h, respectively, the [ 18 The radiochemical purity of the compounds 4 was greater than 95%.

[0144] In summary, there was no significant change in the peak time and peak shape of the marker after 1 and 2 hours of in vitro incubation, indicating 18 The F-labeled substance can maintain good in vitro stability in both physiological saline and mouse serum, which is conducive to further in vivo biological experimental research.

[0145] Example 10 Biodistribution Experiment

[0146] 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. They were cultured in DMEM containing 10% fetal bovine serum and 1% penicillin-streptomycin. Approximately 5×10 6 U87 MG cells were injected subcutaneously 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.

[0147] The marker (100 μL, 3.7 MBq) was injected into nude mice carrying U87MG via the tail vein. Each group of mice (n=3) was killed at 15, 30, 60, and 120 min after injection. Blood, brain, heart, liver, spleen, lung, kidney, muscle, bone, intestine, stomach, and tumor were collected, and the 18F radioactivity was measured by a gamma counter, and then weighed. The uptake of radiotracers in all tissues is shown as the percentage of the injected dose / g±standard deviation (%ID / g±SD). The uptake of all radiotracers is shown as the mean±standard deviation. We [ 18 F]-compound 2 and [ 18 The biodistribution of compound 3 was studied in Balb / c mice bearing U87MG tumors. 18 F]-FAPI-42 is a FAPI-targeted tumor imaging agent that is widely used in clinical trials. 18 The biodistribution of F]-FAPI-42 was studied and compared.

[0148] The groups are as follows:

[0149] Comparative group 1: NOTA-FAPI-42 solution was injected 30 minutes in advance, and then [ 18 F]--FAPI-42, the results are shown in Table 4 (%ID / g, Mean±SD, n=3);

[0150] Inhibition group 1: NOTA-FAPI-42 solution was injected 30 minutes in advance, and then [ 18 F]-Compound 2 results are shown in Table 5 (%ID / g, Mean±SD, n=3);

[0151] Suppression group 2: injection of [ 18 F]-Compound 3 was injected with NOTA-FAPI-42 solution at the same time. The results are shown in Table 6 (%ID / g, Mean±SD, n=3).

[0152] Table 4 18 Biodistribution of [F]-FAPI-42 in U87MG tumor-bearing mice

[0153]

[0154] Table 5 18 Biodistribution results of compound 2 in U87MG tumor-bearing mice

[0155]

[0156]

[0157] Table 618 Biodistribution results of compound 3 in U87MG tumor-bearing mice

[0158]

[0159] In U87MG tumor-bearing mice, at 15, 30, 60, and 120 min after injection, the tumor 18 The absolute uptake of [F]-compound 3 was 6.51±0.22, 4.24±0.65, 3.38±0.25 and 2.54±0.22%ID / g, respectively. 18 F]-FAPI-42 (2.23±0.2±0.48, 1.71±0.53, 0.41±0.09 and 0.22±0.02% ID / g at 15, 30, 60 and 120 min, respectively).

[0160] for[ 18 F]-compound 3, a higher tumor-to-blood ratio was observed at 60 and 120 min, which was significantly higher than that of [ 18 F]-FAPI-42 mice (5.72 vs. 1.14 and 9.84 vs. 1.22, respectively). At 15, 60, and 120 min, the radiotracer of the present application also had higher 18 F]-FAPI-42 tumor-to-muscle ratios (4.02 vs. 1.91, 5.51 vs. 1.29, and 4.86 vs. 1.53, respectively). 18 F]-compound 3 showed a higher tumor-to-brain ratio, far exceeding [ 18 F]-FAPI-42 (28.85 vs. 8.78, 28.41 vs. 8.82, 34.79 vs. 1.58 and 18.50 vs. 3.51). At 30, 60 and 120 min, mice [ 18 The tumor-lung ratio of compound 3 was also higher than that of [ 18 F]-FAPI-42 (2.14 vs. 0.69, 4.35 vs. 0.44, and 3.71 vs. 1.00, respectively). More importantly, at 60 and 120 min, [ 18 F]-tumor ratio of compound 3 18 F]-FAPI-42 tumors performed better (3.50 vs. 0.64 and 3.43 vs. 0.78, respectively).

[0161] In addition, the bone background of the radiotracer of the present application at 15, 30, 60 and 120 min after injection was lower than [ 18F]-FAPI-42 (tumor-to-bone ratio: 1.33 vs. 0.63, 1.65 vs. 0.67, 1.81 vs. 0.28, and 0.89 vs. 0.34, respectively).

[0162] As mentioned above, NOTA-FAPI-42 is a competitive FAP inhibitor with an IC 50 In mice that received NOTA-FAPI-42 (77 μg) and the above radiotracer (blocking studies), tumor uptake was significantly reduced (~91%) (0.31±0.05% ID / g 60 min after injection). Therefore, our biodistribution data, combined with the results of the above in vitro FAP inhibition experiments, suggest that [ 18 The good uptake of compound 3 in U87MG-bearing mice was due to the expression of FAP, while [ 18 F]-compound 3 may be a tumor imaging agent targeting FAP.

[0163] Embodiment 11 18 PET imaging of [F]-compound 3 in tumor-bearing mice

[0164] Will[ 18 F]-Compound 3 (approximately 3.7 MBq) was intravenously injected into mice bearing U87MG tumors. PET imaging was performed 1 h after injection. Mice were anesthetized with about 1.5% concentration of gaseous isoflurane and placed prone on the scanner bed. Image data were processed using OsiriX Mac.

[0165] Inhibition group 3 was a competitive FAP ligand (NOTA-FAPI-42, 77 μg) combined with [ 18 F]-Compound 3 was injected into nude mice carrying U87MG via the tail vein. PET imaging was performed 1 h after injection. At 60 min after injection, [ 18 F]-compound 3 showed good tumor imaging quality ( Figure 4 A). 18 F]-Compound 3 tumor imaging was clear, while uptake in muscle, brain, lung, blood, and liver was almost negligible.

[0166] After blocking with NOTA-FAPI-42 as a competitive FAP inhibitor, [ 18 PET imaging studies of [F]-compound 3 showed poor tumor uptake, indicating that [ 18 The good uptake of [F]-compound 3 in tumors is due to the expression of FAP ( Figure 4 B).

[0167] In the experiment, it was significantly concentrated in the tumor, while in the inhibition group 3, the tumor uptake was significantly reduced, indicating that [ 18 The uptake of [F]-compound 3 in tumors is specific, and the binding of the radioactive drug to the FAP-α receptor can be competitively replaced by active drug precursors at the same target, indicating that it can be used as a new type of PET molecular probe with excellent tumor affinity.

[0168] In the present invention, by simultaneously introducing R1, R2 and other structures into the general formula, the structure of the present application can maintain good stability as a whole, and its water solubility and lipophilicity are better, and it can be used to diagnose or treat diseases characterized by overexpression of fibroblast activation protein (FAP). More importantly, some of the compounds of the present application adopt [ 18 F]AIF radiolabeling strategy is a new type of positron nuclide labeling technology based on [ 18 The formation of [F][AIF]+ cation and its complexation with the bifunctional integrator NOTA has the advantages of one-step labeling, no need for drying and water removal, and simple purification process.

[0169] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. An aluminum fluoride-labeled compound targeting FAP, having a structure represented by general formula (I) or a pharmaceutically acceptable salt thereof, in, X is selected from O or Among them, R 11 is selected from H, alkyl, and haloalkyl; R1 is selected from -CO-NH-, -NH-CO-, -CO-, -NH-, -C(O)O-, -OC(O)-, -OO-, -SS-, C2 to C3 with or without substituents 15 Alkyl, C3~C 15 Heteroalkyl, C3~C 15 Cycloalkyl, C4~C 15 Heterocycloalkyl, C6~C 15 Aryl or C5~C 15 Heteroaryl, C3~C 15 Cycloalkyl-C2~C 15 Alkyl, C4~C 15 Heterocycloalkyl-C2~C 15 Alkyl, C6~C 15 Aryl-C2~C 15 Alkyl, C5~C 15 Heteroaryl-C2~C 15 alkyl. R2 is a group formed by the complexation of the ligand residue with the nuclide, wherein the nuclide is preferably 18 F. 19 F, nuclide is the complex ion AlF 2+ Re-complexed.

2. The aluminum fluoride labeled compound according to claim 1, characterized in that X is selected from O or Among them, R 11 Selected from H, methyl, ethyl, propyl; R1 is selected from Among them, Y is selected from R3 is selected from methyl, ethyl, propyl; Z is selected from -CO-NH-, -NH-CO-, -CO-, and R4 is selected from R5 is selected from q is an integer from 1 to 10; n is an integer from 1 to 10; R2 is a group formed by the complexation of a nuclide with a residue of a NOTA ligand, a DOTA ligand, a DTPA ligand, a HYNIC ligand or a DFO ligand, wherein the nuclide is Al 18 F or Al 19 F.

3. The aluminum fluoride labeled compound according to claim 1, characterized in that X is selected from O or N-CH3; R1 is selected from R2 is a group formed by the complexing nuclide with the residue of NOTA ligand.

4. The aluminum fluoride labeled compound according to claim 1, characterized in that The structure in the general formula (I) Selected from R1 is selected from 5. The aluminum fluoride labeled compound according to claim 4, characterized in that The structure For the following groups:

6. A precursor compound of an aluminum fluoride labeling compound targeting FAP, characterized in that: It has a structure shown in the general formula (I), wherein X is consistent with X defined in any one of claims 1 to 3; R1 is consistent with R1 defined in any one of claims 1 to 3, and R2 is a structure containing an 8-10 membered heterocyclic ring, wherein the heterocyclic ring contains at least one N, preferably containing The structure of , wherein R7, R8, and R9 are independently H, carboxyl, amide, amino, hydroxyl, nitro, or optionally substituted alkyl, alkoxy, or alkylthio. Preferably, R2 is 7. The precursor compound according to claim 6, characterized in that The precursor compound includes any one of the following compounds:

8. A PET imaging agent, characterized in that: The aluminum fluoride labeled compound targeting FAP according to any one of claims 1 to 5 is preferably labeled with Al 18 F.Al 19 F-labeled compounds.

9. Use of the aluminum fluoride-labeled compound targeting FAP according to any one of claims 1 to 5 in the preparation of drugs for radioactive diagnosis and treatment of diseases.

10. Use of the aluminum fluoride labeled compound targeting FAP according to any one of claims 1 to 5 in the preparation of a tumor diagnostic imaging agent.

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