FAP inhibitor

Abstract

To provide a compound for the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP), and a pharmaceutical composition comprising the compound.SOLUTION: A composition comprising a compound having the formula (I) is provided. The composition is useful for the diagnosis or treatment of a disease selected from the group consisting of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and keloid disorders.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to compounds, pharmaceutical compositions comprising or consisting of said compounds, kits comprising or consisting of said compounds or pharmaceutical compositions, and the use of said compounds or pharmaceutical compositions in the diagnosis or treatment of diseases characterized by the overexpression of fibroblast activation protein (FAP). [Background technology]

[0002] Tumor growth and spread are determined not only by cancer cells but also by non-malignant components of malignant lesions, categorized as stroma. In tumors with desmoplastic reactions, such as those of breast, colon, and pancreatic cancer, the stroma can account for more than 90% of the tumor mass. In particular, a subpopulation of fibroblasts, termed cancer-associated fibroblasts (CAFs), is known to be involved in tumor growth, metastasis, and progression. Therefore, these cells are attractive targets for diagnostics and antitumor therapy.

[0003] A distinctive feature of CAFs is the expression of seprase, or fibroblast activation protein α (FAP-α), a type II membrane-bound glycoprotein belonging to the dipeptidyl peptidase 4 (DPP4) family. FAP-α possesses both dipeptidyl peptidase and endopeptidase activities. Endopeptidase activity distinguishes FAP-α from other members of the DPP4 family. Substrates for endopeptidase activity identified to date include denatured type I collagen, α1-antitrypsin, and several neuropeptides. FAP-α plays a role in normal developmental processes during embryogenesis and tissue modeling. It is either not expressed or expressed at very low levels in normal adult tissues. However, it is highly expressed in wound healing, arthritis, atherosclerotic plaques, fibrosis, and over 90% of epithelial cancers.

[0004] The presence of FAP-α in CAFs of many epithelial tumors and the fact that its overexpression is associated with poor prognosis in cancer patients have led to the hypothesis that FAP-α activity is also involved in cancer development, as well as the metastasis and spread of cancer cells. Therefore, targeting this enzyme for imaging and internal radiotherapy could be considered a promising strategy for the detection and treatment of malignant tumors. We developed small molecules based on FAP-α-specific inhibitors and demonstrated specific uptake, rapid internalization, and successful tumor imaging in animal models and tumor patients. A commonly used radioactive tracer, 18 F-fluorodeoxyglucose ( 18 Comparison with F-FDG revealed clear advantages of this new FAP-α ligand in patients with locally advanced lung adenocarcinoma. Thus, the present invention provides, inter alia, (i) detection of smaller primary tumors, thereby enabling earlier diagnosis; (ii) detection of smaller metastases, thereby enabling better tumor staging; (iii) accurate intraoperative guidance facilitating complete surgical removal of tumor tissue; (iv) better differentiation of inflammation from tumor tissue; (v) more accurate staging of tumor patients; (vi) better follow-up of tumor lesions after antitumor therapy; and (vii) the opportunity to use the molecules of the present invention as theranostic (referring to a diagnostic test employing a specific treatment) agents for diagnosis and treatment. The molecules of the present invention can also be used in the diagnosis and treatment of non-malignant diseases such as chronic inflammation, atherosclerosis, fibrosis, tissue remodeling, and keloid disorders. Summary of the Invention

[0005] In a first aspect, the present invention provides a compound of formula (I): [ka] wherein Q, R, U, V, W, Y, and Z are independently present or absent, provided that at least three of Q, R, U, V, W, Y, and Z are present; Q, R, U, V, W, Y and Z are independently O, CH2, NR 4, C=O, C=S, C=NR 4 , HCR 4 and R 4 CR 4 with the proviso that no two O's are immediately adjacent to each other; R 1 and R 2 are independently -H, -OH, halo, C 1-6 -Alkyl, -OC 1-6 -Alkyl, SC 1-6 - selected from the group consisting of alkyl; R 3 is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)-alkyl, -C(O)-aryl, -C=CC(O)-aryl, -C=CS(O)2-aryl, -CO2H, -SO3H, -SON2NH2, -PO3H2 and 5-tetrazolyl; R 4 -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -Alkyl, aryl and -C 1-6 -aralkyl, each -C 1-6 -alkyl is optionally substituted by 1 to 3 substituents selected from -OH, oxo, halo, and may be attached to Q, R, U, V, W, Y, or Z; R 5 -H, halo and C 1-6 - selected from the group consisting of alkyl;

[0006] R 6 and R 7 independently -H, [ka] where R 6 and R 7 is not simultaneously H; where L is a linker; D, A, E and B are independently present or absent, preferably at least A, E and B are present, where, if present: D is a linker; A is NR 4 , O, S, and CH2; E is C 1-6 -alkyl, [ka] selected from the group consisting of: where i is 1, 2, or 3; where j is 1, 2, or 3; where k is 1, 2, or 3; where m is 1, 2 or 3;

[0007] A and E together form a group selected from cycloalkyl, heterocycloalkyl, aryl and heteroaryl, where A and E may be monocyclic, bicyclic and polycyclic, preferably monocyclic; each A and E may be -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each -C 1-6 -Alkyl may be substituted with 1 to 3 substituents selected from -OH, oxo and halo, and may be A, B, D, E or [ka] may be bonded to; B, S, NR 4 , N.R. 4 -O, NR 4 -C 1-6 -Alkyl, NR 4 -C 1-6 -Alkyl-NR 4and 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, the heterocycles preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably NR 4 -C 1-6 -Alkyl-NR 4 and the N-containing heterocycle is C 1-6 -Alkyl, aryl, C 1-6 -substituted by 1 to 3 substituents selected from the group consisting of -aralkyl; and R 8 is selected from the group consisting of a radioactive moiety, a chelating agent, a fluorescent dye, an imaging agent, and combinations thereof; [ka] is a 1-naphthyl moiety or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle has two ring atoms between the N atom and X; and the heterocycle may further contain one, two, or three heteroatoms selected from O, N, and S; and X is a C atom. or a pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof.

[0008] In a second aspect, the present invention relates to a pharmaceutical composition comprising or consisting of at least one compound of the first aspect and a pharmaceutically acceptable carrier and / or excipient.

[0009] In a third aspect, the present invention relates to a compound of the first aspect or a pharmaceutical composition of the second aspect for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in an animal or human subject.

[0010] In a fourth aspect, the present invention relates to a kit comprising or consisting of a compound of the first aspect or a pharmaceutical composition of the second aspect and instructions for diagnosing a disease. [Brief explanation of the drawings]

[0011] The contents of the figures contained in this specification are described below, and in this context reference is made to the detailed description of the invention above and / or below.

[0012] [Figure 1] In vitro characterization of 125I-FAPI-01 and 177Lu-FAPI-02. A. Binding of radiolabeled FAPI-01 and FAPI-02 to various human cancer cell lines, including human FAP-α-transfected cell lines (HT-1080-FAP), murine FAP-α-transfected cell lines (HEK-muFAP), and human CD26-transfected cell lines (HEK-CD26), after 60 minutes of incubation. B. Internalization of radiolabeled FAPI-01 and FAPI-02 into HT-1080-FAP cells after 10 minutes to 24 hours of incubation. The internalized fractions are shown in gray and black, respectively; the extracellular-bound fraction is shown in white. C. Competitive binding of radiolabeled FAPI-01 and FAPI-02 to HT-1080-FAP cells after the addition of increasing concentrations of unlabeled FAPI-01 and Lu-FAPI-02. D. Internalization of FAPI-02 into FAP-α-positive and FAP-α-negative cell lines. Blue: DAPI; Green: FAPI-02-Atto488. E+F. Efflux kinetics of FAPI-01 and FAPI-02 after 1 hour of incubation of HT-1080-FAP cells with radiolabeled compounds followed by 1–24 hours of incubation in compound-free medium. All values ​​are expressed as a percentage of the total applied dose normalized to 1 million cells (%ID / 1 million cells).

[0013] [Figure 2]Binding specificity and relative internalization rates of FAPI derivatives. A-C. Binding and internalization rates of FAPI-03 to FAPI-15 relative to FAPI-02 (defined as 100%). Internalization rates after 1, 4, and 24 hours of incubation are shown in gray; the extracellular bound fraction is shown in white bars. D. Binding of selected FAPI derivatives to HEK cells expressing murine FAP-α and human CD26 after 60 minutes of incubation. Right: binding rates of muFAP and CD26. E. Competitive binding of selected FAPI derivatives to HT-1080-FAP cells after the addition of increasing concentrations of unlabeled compounds.

[0014] [Figure 3] Imaging of FAPI-02 and -04 in mice bearing human FAP-positive (HT-1080-FAP) and -negative (Capan-2, SK-LMS-1) tumor xenografts. A+C, E+G. Small animal PET imaging was performed at the indicated times after intravenous administration of 4 nmol of 68Ga-FAPI-02 and -04 (10 MBq each). The radiotracer does not accumulate in noncancerous tissue but is rapidly concentrated within the tumor (indicated by red arrows). Furthermore, rapid excretion via the kidneys and bladder is observed. B+D, F+H. Quantification of PET images demonstrates solid clearance of 68Ga-FAPI-02 and -04 from the cardiovascular system and consistent uptake into the tumor.

[0015] [Figure 4] Blocking experiments to analyze in vivo binding specificity. A+D. Blocking of tumor accumulation of 68Ga-FAPI-02 and -04 by co-administration of 30 nmol of unlabeled compound to HT-1080-FAP tumor-bearing mice. B+C, E+F. Time activity curves of 68Ga-FAPI-02 and -04 in selected organs after intravenous administration with and without unlabeled compound as competitor.

[0016] [Figure 5]Organ distribution of 177Lu-FAPI-02 and -04 in HT-1080-FAP tumor-bearing nude mice. The biodistribution of AC.177Lu-FAPI-02 and -04 was measured ex vivo at the indicated times after intravenous administration of 1 MBq to mice bearing human FAP-positive HT-1080 tumor xenografts; n=3 per time point. Values ​​are expressed as the percentage of the administered dose per gram of tissue (%ID / g). The radiotracers were shown to accumulate within FAP-expressing tumors, with maximum concentrations observed for FAPI-02 (4.5%ID / g) at 1 hour and FAPI-04 (5.4%ID / g) at 2 hours. DF. Tumor-to-normal tissue ratios were measured 1, 4, and 24 hours after intravenous administration of 177Lu-FAPI-02 and -04.

[0017] [Figure 6] PET / CT imaging of FAPI-02 in cancer patients. 6A-C. Maximum intensity projection (MIP) from a PET / CT scan of a patient with metastatic breast cancer. D. Maximum tissue uptake 10 minutes, 1 hour, and 3 hours after intravenous administration of 68Ga-FAPI-02 to a patient with metastatic breast cancer.

[0018] [Figure 7] PET / CT imaging of FAPI-02 in cancer patients. MIP of PET / CT scans 1 hour after administration of 68Ga-FAPI-02 in patients with pancreatic cancer, non-small cell lung cancer (NSCLC), esophageal cancer, and rectal cancer.

[0019] [Figure 8] PET / CT imaging of FAPI-02 in cancer patients. MIP of PET / CT scan 1 hour after administration of 68Ga-FAPI-02 in a patient with nasopharyngeal and laryngeal cancer.

[0020] [Figure 9]PET / CT imaging of FAPI-02 in cancer patients. 9A+B. Whole-body PET / CT imaging (MIP) 1 hour after administration of 18F-FDG and 68Ga-FAPI-02 in a patient with locally advanced lung adenocarcinoma. C+D. Transaxial images of a patient with lung adenocarcinoma 1 hour after administration of 18F-FDG and 68Ga-FAPI-02. FAPI-02 selectively accumulates in FAP-α-expressing tissues, demonstrating significantly higher uptake in malignant lesions compared to 18F-FDG.

[0021] [Figure 10-16] PET / CT imaging of FAPI-04 in cancer patients 10. Maximum intensity projection (MIP) of PET / CT scans of a metastatic breast cancer patient 10 minutes, 1 hour, and 3 hours after administration of 68Ga-FAPI-04.

[0022] [Figure 11] MIP of PET / CT scans 1 hour after administration of 68Ga-FAPI-04 in patients with sigma carcinoma, hypopharyngeal carcinoma, neuroendocrine tumor, cholangiocarcinoma, ovarian cancer, and small intestine cancer.

[0023] [Figure 12] MIP of PET / CT scan 1 hour after administration of 68Ga-FAPI-04 in a lung cancer patient.

[0024] [Figure 13] MIP of PET / CT scan 1 hour after administration of 68Ga-FAPI-04 in a patient with oncogenic rickets.

[0025] [Figure 14] Comparative imaging of one patient with metastatic prostate cancer. MIP of PET / CT scan 1 hour after application of radiolabeled DOTATATOC, PSMA, and FAPI-04.

[0026] [Figure 15] Maximum intensity projection (MIP) and time radioactivity curves of dynamic 68Ga-FAPI-04 PET / CT scans in a patient with pancreatic cancer.

[0027] [Figure 16] Relative binding rates of Lu-177 labeled FAPI derivatives compared to FAPI-04 (set to 100%) after 1, 4 and 24 hours of incubation on FAP-expressing HT-1080 cells; n=3.

[0028] [Figure 17] Competitive binding of selected FAPI derivatives to HT-1080-FAP cells after addition of increasing concentrations of unlabeled compounds (10-10 to 10-5 M, 60 min incubation, n=3).

[0029] [Figure 18] Binding of FAPI derivatives to HEK cells expressing mouse FAP and human CD26 after 60 min of incubation, n = 3. Values ​​are shown as percentage of applied dose (%ID) per mio cells.

[0030] [Figure 19] Biodistribution of selected FAPI derivatives in HT-1080-FAP xenografts 1, 4, and 24 hours after intravenous administration of the radiotracer, n=3. Values ​​are shown as percentage of the administered dose per gram of tissue (%ID / g).

[0031] [Figure 20] Tumor-to-blood ratios of selected FAPI derivatives in HT-1080-FAP xenografts 1, 4, and 24 hours after intravenous administration of the radiotracer, n=3.

[0032] [Figure 21] PET imaging of Ga-68 labeled FAPI-21 and FAPI-46 in HT-1080-FAP tumor-bearing mice; n=1.

[0033] [Figure 22] Maximum standardized uptake value (SUV) of selected FAPI derivatives in HT-1080-FAP tumor-bearing mice; n=1.

[0034] [Figure 23] Maximum (SUVmax, FIG. 23A) and mean (SUVmean, FIG. 23B) standardized uptake values ​​of Ga-68 labeled FAPI-02 and FAPI-04 in cancer patients; n=25.

[0035] [Figure 24] Intra-individual comparison of six patients with six different tumor entities who underwent FDG-PET and FAPI-PET imaging within nine days.

[0036] [Figure 25] Ga-68 labeled FAPI-04 PET / CT imaging in patients with peritonitis (A), myocarditis (B), and hip joint arthrosis at 1 hour post-injection (1 h pi).

[0037] [Figure 26] PET / CT imaging of Ga-68 labeled FAPI-21 in cancer patients 1 hour after injection (1 h pi)

[0038] [Figure 27] PET / CT imaging of Ga-68 labeled FAPI-46 1 hour post-injection (1h pi) and Sm-153 labeled FAPI-46 intratherapeutical imaging 30 minutes post-injection (30min pi) in cancer patients

[0039] [Figure 28] Therapeutic imaging of Sm-153-labeled FAPI-46 up to 20 hours post-injection (20 h pi)

[0040] [Figure 29]A. Maximum intensity projection (MIP) 1 hour after intravenous administration of 68Ga-FAPI-46 in a patient with metastatic colorectal cancer. B. Bremsstrahlung imaging 2 hours after treatment of the same patient with 90Y-FAPI-46.

[0041] [Figure 30] PET / CT imaging of Ga-68-labeled FAPI-46 1 hour post-injection (1 h pi) in a lung cancer patient with idiopathic pulmonary fibrosis. A, B. Maximum tracer uptake in tumor tissue is significantly higher than that in non-progressing fibrotic lesions. C. Maximum tracer uptake in tumor tissue is slightly lower than that in progressing fibrotic tissue.

[0042] [Figure 31] A. Binding of Tc-99m-labeled FAPI-19 to HT-1080-FAP cells, n=3. B. Binding of Tc-99m-labeled FAPI-19 to HT-1080-FAP cells after the addition of increasing concentrations of unlabeled compound (10-10 to 10-5 M, 60 min incubation, n=3). C. Scintigraphy of Tc-99m-labeled FAPI-19 in HT-1080-FAP xenografts, n=1.

[0043] [Figure 32] A. Binding of Tc-99m-labeled FAPI-34 to HT-1080-FAP cells, n=3. B. Scintigraphy of Tc-99m-labeled FAPI-34 in HT-1080-FAP xenografts, n=1.

[0044] [Figure 33] Scintigraphy of Tc-99m-labeled FAPI-34 in a patient with metastatic pancreatic cancer.

[0045] [Figure 34]A. Binding of Pb-203-labeled FAPI derivatives to HT-1080-FAP cells, n=3. B. Efflux kinetics of Pb-203-labeled FAPI derivatives after 60 min incubation of HT-1080-FAP cells with radiolabeled compounds followed by 1 h incubation in non-radioactive medium, n=3. C. Competitive binding of Pb-203-labeled FAPI to HT-1080-FAP cells after addition of increasing concentrations of unlabeled compounds (10-10 to 10-5 M, 60 min incubation, n=3).

[0046] [Figure 35] Scintigraphy of Pb-203-labeled FAPI-04 and FAPI-46 in HT-1080-FAP xenografts, n = 1.

[0047] [Figure 36] Biodistribution of Pb-203-labeled FAPI-04 and FAPI-46 in HT-1080-FAP xenografts 1, 4, 6, and 24 hours after intravenous administration of the radiotracer, n=3. Values ​​are shown as percentage of injected dose per gram of tissue (%ID / g).

[0048] [Figure 37] A. Binding of Cu-64-labeled FAPI-42 and FAPI-52 to HT-1080-FAP cells, n=3. B. Competitive binding of Cu-64-labeled FAPI-42 and FAPI-52 to HT-1080-FAP cells after the addition of increasing concentrations of unlabeled compounds (10-10 to 10-5 M, 60 min incubation, n=3). C. Efflux kinetics of Cu-64-labeled FAPI-42 and FAPI-52 after incubation of HT-1080-FAP cells with radiolabeled compounds for 60 min, followed by 1 to 24 h incubation in non-radioactive medium, n=3.

[0049] [Figure 38] PET imaging of Cu-64 labeled FAPI-42 and FAPI-52 in HT-1080-FAP tumor-bearing mice; n=1.

[0050] [Figure 39] PET imaging of AlF-18 labeled FAPI-42 and FAPI-52 in HT-1080-FAP tumor-bearing mice; n=1.

[0051] [Figure 40] a. Small animal PET imaging of 68Ga-labeled FAPI-02 in U87MG tumor-bearing nude mice up to 140 min after intravenous administration of the radiotracer. The tumor is indicated by the red arrow. b. Biodistribution of 177Lu-labeled FAPI-02 and FAPI-04 in U87MG tumor-bearing nude mice 1, 4, and 24 h after intravenous administration of the radiotracer; n=3.

[0052] [Figure 41] Tumor-to-organ ratios of 177Lu-labeled FAPI-02 and -04 in U87MG tumor-bearing mice 1, 4, and 24 hours after intravenous administration.

[0053] [Figure 42] Maximum intensity projection (MIP) of a PET / CT scan in a patient with glioblastoma 10 minutes, 1 hour, and 3 hours after administration of 68Ga-FAPI-02.

[0054] [Figure 43] Exemplary images of IDHwt glioblastoma, IDH-mutated glioma WHO grade II, and IDH-mutated glioblastoma (contrast-enhanced T1-weighted MRI, FAPI-PET, and fusion images of both modalities).

[0055] [Figure 44] Absolute SUVmax values ​​for all 18 types of glioma.

[0056] [Figure 45]Statistical analysis of SUVmax / BG values. Boxplots and corresponding ROC curves of SUVmax / BG values ​​in GBM vs. non-GBM (a, b), IDH mutant vs. IDH wild-type gliomas (c, d), and glioma grade II vs. glioma grade III / IV (e, f).

[0057] [Figure 46] Dose-dependent inhibition of enzymatic FAP activity by FAPI-04 and talabostat. In contrast to talabostat, a potent DPP4 inhibitor with marginal FAP activity, FAPI-04 demonstrates robust dose-dependent FAP inhibition.

[0058] [Figure 47] Reuptake of 177Lu-labeled FAPI-04 and FAPI-46 in HT-1080-FAP cells. After incubating cells with the radiotracer at 37°C for 60 minutes, the compound was removed and nonradioactive medium with (+Comp.) or without (-Comp.) the unlabeled compound was added and incubated for 10 minutes to 6 hours. Already within the first 10 minutes of incubation, new uptake of the unlabeled FAPI derivative occurred, displacing a portion of the radiolabeled fraction, resulting in a significant decrease in radioactivity compared to pure medium without competitor. After 6 hours of incubation, nearly complete displacement of the radiolabeled FAPI had occurred. These findings indicate the continued reuptake of intact FAP molecules that returned to the cell membrane during initial internalization, allowing for renewed binding and internalization of the FAP ligand.

[0059] [Figure 48] Organ distribution of 177Lu-FAPI-04 after single and multiple injections in HT-1080-FAP tumor-bearing nude mice. Two doses of equal doses of 177Lu-FAPI-04, administered 4 hours apart, resulted in increased overall organ activity, including tumor activity, measured 8 and 24 hours after the first injection. In contrast, three doses (higher initial dose followed by lower subsequent doses) revealed no change in overall organ activity.

[0060] [Figure 49] Binding of F-18-FAPI derivatives to human FAP-expressing HT1080 cells after 10, 30, 60, and 90 minutes of incubation, n=3. Values ​​are shown as percentage of applied amount per miocell (%ID).

[0061] [Figure 50] PET imaging of AlF-18 labeled FAPI-74 and FAPI-52 in HT-1080-FAP tumor-bearing mice; n=1.

[0062] [Figure 51] Biodistribution of FAPI-75 in HT-1080-FAP xenografts 1, 4, and 24 hours after intravenous administration of the radiotracer, n=3. Values ​​are presented as percentage of the administered dose per gram of tissue (%ID / g).

[0063] [Figure 52] PET imaging of patients with non-small cell lung cancer: robust accumulation of F18-labeled FAPI-74 in multiple metastases.

[0064] [Figure 53] Time radioactivity curves of cardiac region (SUVmean) for FAPI-04 and -46 as an illustration of fast blood pool clearance.

[0065] [Figure 54] FAPI-02 and FAPI-04 at different imaging time points (10 min, 1 h, and 3 h after injection) in two patients with metastatic breast cancer. Rapid tumor targeting and rapid blood clearance are followed by a long plateau phase with no associated changes in image contrast (top). Compared to FAPI-02, the ligand FAPI-04 is characterized by a prolonged tumor retention time (bottom).

[0066] [Figure 55]The effective dose of FAPI-02 was 1.80E-02 mSv / MBq calculated by OLINDA (1.82E-02 by IDAC1 / ICRP60 and 1.79E-02 by IDAC2 / ICRP103). The effective dose of FAPI-04 PET / CT was 1.64E-02 mSv / MBq calculated by OLINDA (1.66E-02 by IDAC1 / ICRP60 and 1.35E-02 by IDAC2 / ICRP103). If the delayed scan 3 hours after injection is omitted in the clinical setting, the routine activity of the FAPI examination can be reduced to 200 MBq of 68Ga, and the radiation dose of such FAPI-PET / CT scans will be continuously 3 - 4 mSv.

[0067] [Figure 56] A) 68Ga-FAPI-04 one hour after injection into different tumor entities in PETCT. The highest mean SUVmax (>12) was found in sarcoma, esophageal cancer, breast cancer, cholangiocarcinoma and lung cancer. The lowest FAPI uptake (mean SUVmax < 6) was observed in renal cell carcinoma, differentiated thyroid cancer, adenoid cystic carcinoma, gastric cancer and pheochromocytoma. The mean SUVmax of hepatocellular carcinoma, colorectal cancer, head and neck cancer, ovarian cancer, pancreatic cancer was intermediate (6 < x < 12). High inter-individual variability was observed within all tumor entities. Due to the low background radioactivity (SUV2), the tumor-to-background ratio was >2-fold in the intermediate uptake group and >4-fold in the high-intensity uptake group. B) In the primary tumor entities, similar SUV uptake was shown compared to the tumor entities using FAPI-04. <000​​​​​​​​​​​Before describing the present invention in detail below, it will be understood that this invention is not limited to the particular method protocols and reagents described herein, as these may vary. It is also understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is limited only by the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0070] Preferably, the terms used herein are defined as set forth in "A multilingual glossary of biotechnological terms: (IUPAC Recommendations)", Leuenberger, HGW, Nagel, B. and Klbl, H. eds. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland).

[0071] Throughout this specification and the claims that follow, unless the context requires otherwise, the terms "comprise" and variations such as "comprises" and "comprising" will be understood to mean the inclusion of a stated integer or step or group of integers or steps, and not the exclusion of other integers or steps or group of integers or steps. The following passages define various aspects of the invention in more detail. Each aspect so defined may be combined with other aspects or aspects, unless expressly indicated to the contrary. In particular, any feature indicated as optional, preferred, or advantageous may be combined with other features or features indicated as optional, preferred, or advantageous.

[0072] Several documents are cited throughout the text of this specification. Each document cited herein (including all patents, patent applications, scientific publications, manufacturer's specifications, instructions, etc.), whether supra or infra, is hereby incorporated by reference in its entirety. Nothing herein should be construed as an admission that the present invention is not entitled to antedate such disclosure by virtue of prior invention. Some of the documents cited herein are characterized as "incorporated by reference." In the event of a conflict between a definition or teaching of such incorporated document and a definition or teaching cited herein, the text of this specification shall control.

[0073] The elements of the present invention are described below. While these elements are listed in specific embodiments, it should be understood that they may be combined in any manner and in any number to create further embodiments. The various described examples and preferred embodiments should not be construed as limiting the invention to only those embodiments explicitly described. The specification should be understood to support and encompass embodiments that combine the explicitly described embodiments with any disclosed and / or preferred elements. Furthermore, any permutation and combination of all elements described herein should be considered disclosed by the description of this application unless the context indicates otherwise.

[0074] definition Below are definitions of some commonly used terms used herein, which, in each instance of their use, have their respective defined and preferred meanings in the remainder of the specification.

[0075] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the content clearly dictates otherwise.

[0076] Provided below are definitions of the terms alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, alkenyl, and alkynyl, which have their respective defined and preferred meanings in each instance as used in the remainder of this specification.

[0077] The term "alkyl" refers to a saturated, straight or branched carbon chain. Preferably, the chain contains 1 to 10 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms (e.g., methyl, ethylmethyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, pentyl, or octyl). Alkyl groups may be substituted.

[0078] The term "heteroalkyl" refers to a saturated, straight or branched carbon chain. Preferably, the chain contains 1 to 9, i.e., 1, 2, 3, 4, 5, 6, 7, 8, or 9, carbon atoms (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, octyl), which is interrupted one or more times, e.g., 1, 2, 3, 4, or 5 times, by the same or different heteroatoms. Preferably, heteroatoms are selected from O, S and N, such as -O-CH, -S-CH, -CH-O-CH, -CH-O-C2H, -CH-S-CH, -CH-S-C2H, -C2H-O-CH, -C2H-O-C2H, -C2H-S-CH, -C2H-S-C2H, etc. Heteroalkyl groups may be substituted.

[0079] The terms "cycloalkyl" and "heterocycloalkyl," by themselves or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl," respectively, preferably having 3, 4, 5, 6, 7, 8, 9, or 10 atoms forming the ring, e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The terms "cycloalkyl" and "heterocycloalkyl" are also meant to include bicyclic, tricyclic, and polycyclic versions thereof. The term "heterocycloalkyl" preferably refers to a 5-membered saturated ring (at least one member is an N, O, or S atom, and may contain one additional O or one additional N); a 6-membered saturated ring (at least one member is an N, O, or S atom, and may contain one additional O, one additional N, or two additional N atoms); or a 9- or 10-membered saturated bicyclic ring (at least one member is an N, O, or S atom, and may contain one additional N atom, two additional N, or three additional N atoms). "Cycloalkyl" and "heterocycloalkyl" groups may be substituted. Furthermore, in the case of heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, spiro[3,3]heptyl, spiro[3,4]octyl, spiro[4,3]octyl, spiro[3,5]nonyl, spiro[5,3]nonyl, spiro[3,6]decyl, spiro[6,3]decyl, spiro[4,5]decyl, spiro[5,4]decyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, and the like.Examples of heterocycloalkyl include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, 1,8 diazo-spiro-[4,5]decyl, 1,7 diazo-spiro-[4,5]decyl, 1,6 diazo-spiro-[4,5]decyl, 2,8 diazo-spiro[4,5]decyl, 2,7 diazo-spiro[4,5]decyl, 2,6 diazo-spiro[4,5]decyl, 1,8 diazo-spiro-[5,4]decyl, 1,7 diazo-spiro-[5,4]decyl, 2,8 diazo-spiro-[5,4]decyl, 2,7 diazo-spiro[5,4]decyl, 3,8 These include diazo-spiro[5,4]decyl, 3,7 diazo-spiro[5,4]decyl, 1-azo-7,11-dioxo-spiro[5,5]undecyl, 1,4-diazabicyclo[2.2.2]oct-2-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, and 2-piperazinyl.

[0080] The term "aryl" refers to an aromatic monocyclic ring preferably containing 6 carbon atoms, an aromatic bicyclic ring system containing 10 carbon atoms, or an aromatic tricyclic ring system containing 14 carbon atoms. Examples include phenyl, naphthyl, or anthracenyl. Aryl groups may be substituted.

[0081] The term "aralkyl" refers to an alkyl moiety substituted by an aryl, where alkyl and aryl have the meanings outlined above. An example is the benzyl group. Preferably, in this context, the alkyl chain contains 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms (e.g., methyl, ethylmethyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butenyl, tert-butyl, pentyl, hexyl, pentyl, octyl). The aralkyl group may be substituted in the alkyl and / or aryl portions of the group.

[0082] The term "heteroaryl" preferably refers to a 5- or 6-membered aromatic monocyclic ring in which at least one of the carbon atoms is replaced by 1, 2, 3, or 4 (in the case of a 5-membered ring) or 1, 2, 3, 4, or 5 (in the case of a 6-membered ring) heteroatoms, which may be the same or different, preferably selected from O, N, and S; an aromatic bicyclic ring system in which 1, 2, 3, 4, 5, or 6 carbon atoms out of 8, 9, 10, 11, or 12 carbon atoms are replaced by the same or different heteroatoms, preferably selected from O, N, and S; or an aromatic tricyclic ring system in which 1, 2, 3, 4, 5, or 6 carbon atoms out of 13, 14, 15, or 16 carbon atoms are replaced by the same or different heteroatoms, preferably selected from O, N, and S. Examples include oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,5-thiadiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1-benzofuranyl, 2-benzofuranyl, indoyl, isoindoyl, and the like. isoindoyl, benzothiophenyl, 2-benzothiophenyl, 1H-indazolyl, benzimidazolyl, benzoxazolyl, indoxazinyl, 2,1-benzoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl, 2,1-benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, quinolinyl, 1,2,3-benzotriazinyl or 1,2,4-benzotriazinyl.

[0083] The term "heteroaralkyl" refers to an alkyl moiety substituted by a heteroaryl, where alkyl and heteroaryl have the meanings outlined above. Examples include 2-alkylpyridinyl, 3-alkylpyridinyl, or 2-methylpyridinyl. Preferably, in this context, the alkyl chain contains 1 to 8 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms (e.g., methyl, ethylmethyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butenyl, tert-butyl, pentyl, hexyl, pentyl, octyl). Heteroaralkyl groups may be substituted by alkyl and / or heteroaryl groups.

[0084] "Alkenyl" and "cycloalkenyl" refer to an olefinically unsaturated carbon atom containing chain or ring with one or more double bonds. Examples include propenyl and cyclohexenyl. Preferably, the alkenyl chain contains 2 to 8 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, or 8 carbon atoms (e.g., ethenyl, 1-propenyl, 2-propenyl, iso-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, iso-butenyl, sec-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, hexenyl, pentenyl, octenyl). Preferably, the cycloalkenyl ring contains 3 to 8, i.e., 3, 4, 5, 6, 7 or 8 carbon atoms (e.g., 1-cyclopropenyl, 2-cyclopropenyl, 1-cyclobutenyl, 2-cyclobutenyl, 1-cyclopentenyl, 2-cyclopentenyl, 3-cyclopentenyl, cyclohexenyl, cyclopentenyl, cyclooctenyl).

[0085] The term "alkynyl" refers to an unsaturated carbon atom containing chain or ring with one or more triple bonds. An example is the propargyl group. Preferably, the alkynyl chain contains 2 to 8 carbon atoms, i.e., 2, 3, 4, 5, 6, 7, or 8 carbon atoms (e.g., ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, hexynyl, pentynyl, octynyl).

[0086] In one embodiment, carbon or hydrogen atoms in an alkyl, heteroalkyl, cycloalkyl, aryl, aralkyl, alkenyl, cycloalkenyl, or alkynyl group may be substituted, independently of one another, with one or more elements selected from the group consisting of O, S, and N, or with a group containing one or more elements selected from the group consisting of O, S, and N.

[0087] Embodiments include alkoxy, cycloalkoxy, aryloxy, aralkoxy, alkenyloxy, cycloalkenyloxy, alkynyloxy, alkylthio, cycloalkylthio, arylthio, aralkylthio, alkenylthio, cycloalkenylthio, alkynylthio, alkylamino, cycloalkylamino, arylamino, aralkylamino, alkenylamino, cycloalkenylamino, and alkynylamino groups.

[0088] Other embodiments include hydroxyalkyl, hydroxycycloalkyl, hydroxyaryl, hydroxyaralkyl, hydroxyalkenyl, hydroxycycloalkenyl, hydroxyalinyl, mercaptoalkyl, mercaptocycloalkyl, mercaptoaryl, mercaptoaralkyl, mercaptoalkenyl, mercaptocycloalkenyl, mercaptoalkynyl, aminoalkyl, aminocycloalkyl, aminoaryl, aminoaralkyl, aminoalkenyl, aminocycloalkenyl, and aminoalkynyl groups.

[0089] In another embodiment, the hydrogen atoms in the alkyl, heteroalkyl, cycloalkyl, aryl, aralkyl, alkenyl, cycloalkenyl, and alkynyl groups can be substituted, independently of one another, with one or more halogen atoms. One group is a trifluoromethyl group.

[0090] Where two or more groups or two or more residues may be selected independently of each other, the term "independently" means that the groups or residues may be the same or different.

[0091] As used herein, phrases defining a length range limit, such as "1 to 6," refer to any integer between 1 and 6, i.e., 1, 2, 3, 4, 5, and 6. In other words, any range defined by two explicitly stated integers is meant to include and disclose any integers defining that limit, as well as any integers subsumed within that range.

[0092] As used herein, the term "halo" refers to a halogen radical selected from the group consisting of F, Br, I, and Cl. Preferably, the halogen is F.

[0093] The term "linker" as used herein refers to any chemically suitable linker. Preferably, the linker is not cleaved or is cleaved only slowly under physiological conditions. Thus, preferably, the linker does not contain a recognition sequence for proteases or other degradative enzymes. Since the compounds of the present invention are preferably administered systemically, allowing broad access to all compartments in the body, and subsequently concentrating the compounds of the present invention wherever tumors are located in the body, it is preferable that the linker is selected so that it is not cleaved or is cleaved only slowly in the blood. Cleavage is considered slow if less than 50% of the linker is cleaved 2 hours after administration of the compound to a human patient. Suitable linkers include or consist of, for example, optionally substituted alkyl, heteroalkyl, cycloalkyl, cycloheteroalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, sulfonyl, amines, ethers, thioethers, phosphines, phosphoramidates, carboxamides, esters, imidoesters, amidines, thioesters, sulfonamides, 3-thiopyrrolidine-2,5-diones, carbamates, ureas, guanidines, thioureas, disulfides, oximes, hydrazines, hydrazides, hydrazones, diaza bonds, triazoles, triazolines, tetrazines, platinum complexes, and amino acids, or combinations thereof. Preferably, the linker comprises or consists of 1,4-piperazine, 1,3-propane and phenolic ethers or combinations thereof.

[0094] The expression "optionally substituted" refers to a group in which one, two, three or more hydrogen atoms may be replaced, independently of one another, by a respective substituent.

[0095] As used herein, the term "amino acid" refers to any organic acid, aliphatic carboxylic acid derivative, containing one or more amino substituents, e.g., α-, β-, or γ-amino. In the polypeptide notation used herein, e.g., Xaa5, i.e., Xaa1Xaa2Xaa3Xaa4Xaa5, Xaa1 through Xaa5 are each independently selected from defined amino acids, with the left-hand direction being the amino-terminal direction and the right-hand direction being the carboxy-terminal direction, in accordance with standard usage and convention.

[0096] The term "conventional amino acids" refers to the 20 naturally occurring amino acids, including all their stereoisomers, i.e., D,L-, D-, and L-amino acids. These conventional amino acids may be referred to herein by their conventional three-letter or one-letter abbreviations, which abbreviations follow conventional usage (see, e.g., Immunology—A Synthesis, 2nd Edition, E.S. Golub and D.R. Gren, Eds., Sinauer Associates, Sunderland Mass. (1991)).

[0097] The term "unconventional amino acid" refers to unnatural amino acids or chemical amino acid analogs, such as α,α-disubstituted amino acids, N-alkyl amino acids, homoamino acids, dehydroamino acids, aromatic amino acids (other than phenylalanine, tyrosine, and tryptophan), and ortho-, meta-, or para-aminobenzoic acid. Unconventional amino acids also include compounds in which the amine and carboxyl functional groups are separated by one, three, or more substitution patterns, such as β-alanine, γ-aminobutyric acid, Freidinger lactam, bicyclic dipeptide (BTD), amino-methylbenzoic acid, and other compounds commonly known in the art. Stathine-like isosteres, hydroxyethylene isosteres, reduced amide-linked isosteres, thioamide isosteres, urea isosteres, carbamate isosteres, thioether isosteres, vinyl isosteres, and other amide-linked isosteres commonly known in the art can also be used. The use of analogs or non-conventional amino acids can improve the stability and biological half-life of the added peptides by increasing their resistance to degradation under physiological conditions. Those skilled in the art will recognize that similar types of substitutions can be made. A non-limiting list of non-conventional amino acids that can be used as suitable building blocks of peptides and their standard abbreviations (in parentheses) is as follows: α-aminobutyric acid (Abu), LN-methylalanine (Nmala), α-amino-α-methylbutyrate (Mgabu), LN-methylarginine (Nmarg), aminocyclopropane (Cpro), LN-methylasparagine (Nmasn), carboxylate LN-methylaspartic acid (Nmasp), aminoisobutyric acid (Aib), LN-methylcysteine ​​(Nmcys), aminonorbornyl (Norb), LN-methylglutamine (Nmgln), carboxylate LN-methylglutamic acid (Nmglu), cyclohexylalanine (Chexa), LN-methylhistidine (Nmhis), cyclopentylalanine (Cpen), LN-methylisoleucine (Nmile), LN-methylleucine (Nmleu), LN-methyllysine (Nmlys),LN-methylmethionine (Nmmet), LN-methylnorleucine (Nmnle), LN-methylnorvaline (Nmnva), LN-methylornithine (Nmorn), LN-methylphenylalanine (Nmphe), LN-methylproline (Nmpro), LN-methylserine (Nmser), LN-methylthreonine (Nmthr), LN-methyltryptophan (Nmtrp), D-ornithine (Dorn), LN-methyltyrosine (Nmtyr), LN-methylvaline (Nmval), LN-methylethylglycine (Nmetg), LN -methyl-t-butylglycine (Nmtbug), L-norleucine (NIe), L-norvaline (Nva), α-methyl-aminoisobutyrate (Maib), α-methyl-γ-aminobutyrate (Mgabu), D-α-methylalanine (Dmala), α-methylcyclohexylalanine (Mchexa), D-α-methylarginine (Dmarg), α-methylcyclopentylalanine (Mcpen), D-α-methylasparagine (Dmasn), α-methyl-α-naphthalene naphthylalanine N-amino-α-methylaspartate (Dmasp), α-methylpenicillamine (Mpen), D-α-methylcysteine ​​(Dmcys), N-(4-aminobutyl)glycine (NgIu), D-α-methylglutamine (Dmgln), N-(2-aminoethyl)glycine (Naeg), D-α-methylhistidine (Dmhis), N-(3-aminopropyl)glycine (Norn), D-α-methylisoleucine (Dmile), N-amino-α-methylbutyrate (Nmaabu), D-α-methylleucine (Dmleu), α -napthylalanine (Anap), D-α-methyllysine (Dmlys), N-benzylglycine (Nphe), D-α-methylmethionine (Dmmet), N-(2-carbamylethyl)glycine (NgIn), D-α-methylornithine (Dmorn), N-(carbamylmethyl)glycine (Nasn), D-α-methylphenylalanine (Dmphe), N-(2-carboxyethyl)glycine (NgIu), D-α-methylproline (Dmpro), N-(carboxymethyl)glycine (Nasp), D-α-methylserine (Dmser),N-cyclobutylglycine (Ncbut), D-α-methylthreonine (Dmthr), N-cycloheptylglycine (Nchep), D-α-methyltryptophan (Dmtrp), N-cyclohexylglycine (Nchex), D-α-methyltyrosine (Dmty), N-cyclodecylglycine (Ncdec), D-α-methylvaline (Dmval), N-cyclododecylglycine (Ncdod), DN-methylalanine (Dnmala), N-cyclooctylglycine (Ncoct), DN-methylarginine (Dnmarg), N-cyclohexylglycine (Ncbut), N-cyclohexylglycine (Ncdec ... Chloropropylglycine (Ncpro), DN-methylasparagine (Dnmasn), N-cycloundecylglycine (Ncund), DN-methylaspartate (Dnmasp), N-(2,2-diphenylethyl)glycine (Nbhm), DN-methylcysteine ​​(Dnmcys), N-(3,3-diphenylpropyl)glycine (Nbhe), DN-methylglutamine (Dnmgln), N-(3-guanidinopropyl)glycine (Narg), DN-methylglutamate (Dnmglu), N-(1-hydroxyethyl)glycine ( Ntbx), DN-methylhistidine (Dnmhis), N-(hydroxyethyl)glycine (Nser), DN-methylisoleucine (Dnmile), N-(imidazolylethyl)glycine (Nhis), DN-methylleucine (Dnmleu), N-(3-indolylethyl)glycine (Nhtrp), DN-methyllysine (Dnnilys), N-methyl-γ-aminobutyrate (Nmgabu), N-methylcyclohexylalanine (Nmchexa), DN-methylmethionine (Dnmmet), DN-methylornithine (D nmorn), N-methylcyclopentylalanine (Nmcpen), N-methylglycine (NaIa), DN-methylphenylalanine (Dnmphe), N-methylaminoisobutyrate (Nmaib), DN-methylproline (Dnmpro), N-(1-methylpropyl)glycine (Nile), DN-methylserine (Dnmser), N-(2-methylpropyl)glycine (Nleu), DN-methylthreonine (Dnmthr), DN-methyltryptophan (Dnmtrp), N-(1-methylethyl)glycine (Nval),DN-methyltyrosine (Dnmtyr), N-methyl-α-napthylalanine (Nmanap), DN-methylvaline (Dnmval), N-methylpenicillamine (Nmpen), γ-aminobutyric acid (Gabu), N-(p-hydroxyphenyl)glycine (Nhtyr), L- / -butylglycine (Tbug), N-(thiomethyl)glycine (Ncys), L-ethylglycine (Etg), penicillamine (Pen), L-homophenylalanine (Hphe), L-α-methylalanine (Mala), L-α -methylarginine (Marg), L-α-methylasparagine (Masn), L-α-methylaspartate (Masp), L-α-methyl-t-butylglycine (Mtbug), L-α-methylcysteine ​​(Mcys), L-methylethylglycine (Metg), L-α-methylglutamine (MgIn), L-α-methylglutamate (MgIu), L-α-methylhistidine (Mhis), L-α-methylhomophenylalanine (Mhphe), L-α-methylisoleucine (Mile), N-( 2-methylthioethyl)glycine (Nmet), L-α-methylleucine (Mleu), L-α-methyllysine (Mlys), L-α-methylmethionine (Mmet), L-α-methylnorleucine (MnIe), L-α-methylnorvaline (Mnva), L-α-methylornithine (Morn), L-α-methylphenylalanine (Mphe), L-α-methylproline (Mpro), L-α-methylserine (Mser), L-α-methylthreonine (Mthr), L-α-methyltryptophan (M trp), L-α-methyltyrosine (Mtyr), L-α-methylvaline (Mval), LN-methylhomophenylalanine (Nmhphe), N-(N-(2,2-diphenylethyl)carbamylmethyl)glycine (Nnbhm), N-(N-(3,3-diphenylpropyl)carbamylmethyl)glycine (Nnbhe), 1-carboxy-1-(2,2-diphenylethylamino)cyclopropane (Nmbc), LO-methylserine (Omser), LO-methylhomoserine (Omhser).

[0098] As used herein, the term "N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle" means a cyclic saturated or unsaturated hydrocarbon compound containing at least one nitrogen atom as a member of the cyclic chain.

[0099] As used herein, the term "radioactive moiety" refers to a molecular assembly bearing a radioactive nuclide. The nuclide is attached by either a covalent or coordinate bond that remains stable under physiological conditions. Examples include: 131 I]-3-iodobenzoic acid or 68 There's Ga-DOTA.

[0100] As used herein, a "fluorescent isotope" emits electromagnetic radiation after being excited by electromagnetic radiation of a shorter wavelength.

[0101] As used herein, a "radioisotope" is a radioactive isotope of an element (included by the term "radionuclide") that emits α-, β-, and / or γ-radiation.

[0102] The term "radiopharmaceutical" is used in the context of the present invention to refer to biologically active compounds modified with a radioisotope. In particular, intercalating agents can be used to deliver radioactivity directly into the DNA (e.g., Hoechst-33258). 131 I-supported derivatives).

[0103] The terms "chelating agent" and "chelate" are used interchangeably in the context of the present invention and refer to a molecule, often organic, and often a Lewis base, that has two or more unshared electron pairs available for donation to a metal ion. The metal ion is usually coordinated to the chelating agent with two or more electron pairs. The terms "bilidate chelating agent," "triligand chelating agent," and "tetraligand chelating agent" refer to chelating agents that have two, three, and four electron pairs readily available, respectively, for simultaneous donation to the metal ion coordinated by the chelating agent. Typically, the electron pairs of a chelating agent form coordinate bonds with a single metal ion, but in certain instances, a chelating agent may form coordinate bonds with more than one metal ion, and various bonding modes are possible.

[0104] The term "fluorochrome" as used in the context of the present invention is used to refer to a compound that emits visible or infrared light after being excited by electromagnetic radiation of a shorter, suitable wavelength. It is understood by those skilled in the art that each fluorescent dye has a predetermined excitation wavelength.

[0105] The term "contrast agent" is used in the context of the present invention to refer to a compound that increases the contrast of a structure or fluid in medical imaging. The enhancement is achieved by absorbing electromagnetic radiation or by modifying the electromagnetic field.

[0106] As used herein, the term "paramagnetic" refers to paramagnetism induced by unpaired electrons in a medium. Paramagnetic materials induce a magnetic field when an external magnetic field is applied. Unlike diamagnetism, the direction of the induced magnetic field is the same as the external magnetic field, and unlike ferromagnetism, the magnetic field is not maintained in the absence of an external magnetic field.

[0107] The term "nanoparticles" as used herein preferably refers to spherical particles with diameters between 1 and 100 nanometers. Depending on their composition, nanoparticles can have appreciable magnetic, optical, or physicochemical qualities. Furthermore, many types of nanoparticle surface modifications are achievable.

[0108] The term "pharmaceutically acceptable salt" refers to a salt of the compound of the present invention. Suitable pharmaceutically acceptable salts of the compounds of the present invention include, for example, acid addition salts that can be formed by mixing a solution of choline or its derivative with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid, or phosphoric acid. Furthermore, when the compound of the present invention has an acidic moiety, suitable pharmaceutically acceptable salts thereof include alkali metal salts (e.g., sodium salt or potassium salt); alkaline earth metal salts (e.g., calcium salt or magnesium salt); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium, and amine cations formed with counter anions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl sulfonates, and aryl sulfonates). Examples of pharmaceutically acceptable salts include, but are not limited to, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, clavulanate, cyclopentanepropionate, digluconate, dihydrochloride, dodecyl sulfate, edetate, edisylate, estolate, esylate, ethanesulfonate, formate, fumarate, and gluceptate. Gluceptate, glucoheptonate, gluconate, glutamate, glycerophosphate, glycolylarsanilate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonic acid, hydroxynaphthoate, iodide, isothiocyanate, lactate, lactobionate, laurate,Lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methyl sulfate, mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate / diphosphate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, acetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, undecanoate, valerate, etc. (e.g., Berge, SM, et al., (See "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19.) Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into either base or acid addition salts.

[0109] The neutral forms of the compounds may be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound may differ from the various salt forms in certain physical properties, such as solubility in polar solvents, but the salts are otherwise equivalent to the parent form of the compound for purposes of this invention.

[0110] In addition to salt forms, the present invention provides compounds in the form of prodrugs. Prodrugs of the compounds described herein are compounds that readily undergo chemical changes under physiological conditions to provide the compounds of Formula (I). Prodrugs are active or inactive compounds that are chemically modified into compounds of the present invention through in vivo physiological action, such as hydrolysis or metabolism, after the prodrug is administered to a patient. Furthermore, prodrugs can be converted to compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted to compounds of the present invention when placed in a transdermal patch reservoir with an appropriate enzyme. The suitability and techniques involved in the preparation and use of prodrugs are well known to those skilled in the art. For a general discussion of prodrugs involving esters, see Svensson and Tunek, Drug Metabolism Reviews 16.5 (1988) and Bundgaard, Design of Prodrugs, Elsevier (1985). Examples of masked carboxylate anions include various esters, such as alkyl (e.g., methyl, ethyl), cycloalkyl (e.g., cyclohexyl), aralkyl (e.g., benzyl, p-methoxybenzyl), and alkylcarbonyloxyalkyl (e.g., pivaloyloxymethyl). Amines have been masked as arylcarbonyloxymethyl-substituted derivatives, which are cleaved in vivo by esterases to release the free drug and formaldehyde (Bungaard J. Med. Chem. 2503 (1989)). Drugs containing acidic NH groups, such as imidazoles, imides, and indoles, have been masked with N-acyloxymethyl groups (Bundgaard Design of Prodrugs, Elsevier (1985)). Hydroxyl groups have been masked as esters and ethers. EP 0 0 039 051 (Sloan and Little, Apr. 11, 1981) discloses Mannich-base hydroxamic acid prodrugs, their preparation and use.

[0111] Compounds according to the present invention can be synthesized by one or more of the following methods. It should be noted that the general procedures are presented in connection with the preparation of compounds of unspecified stereochemistry. However, such procedures are generally applicable to compounds of a particular stereochemistry, e.g., where the stereochemistry about a group is (S) or (R). Furthermore, compounds of one stereochemistry (e.g., (R)) can often be utilized to prepare compounds of the opposite stereochemistry (i.e., (S)) by well-known methods, e.g., inversion.

[0112] Certain compounds of the present invention can exist in non-solvated form as well as solvated form, including hydrated form.In general, solvated form is equivalent to non-solvated form and is intended to be included within the scope of the present invention.Certain compounds of the present invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent to the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0113] Certain compounds of the present invention possess asymmetric carbon atoms (optical centers) or double bonds; the racemates, diastereomers, geometric isomers and individual isomers are all intended to be encompassed within the scope of the present invention.

[0114] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain unnatural proportions of atomic isotopes, such as tritium ( 3 H), iodine-125( 125 I) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

[0115] As used in this application, the term "pharmaceutical composition" refers to a substance and / or combination of substances used for the identification, prevention, or treatment of a tissue condition or disease. A pharmaceutical composition is formulated so as to be suitable for administration to a patient to prevent and / or treat a disease. Furthermore, a pharmaceutical composition refers to a combination of an active agent, inert or active carrier, making the composition suitable for therapeutic use. Depending on its chemical and physical properties, a pharmaceutical composition may be administered orally, parenterally, topically, inhalatively, rectally, sublingually, transdermally, subcutaneously, or vaginally. route The pharmaceutical compositions can be formulated for use in a pharmaceutical composition. Pharmaceutical compositions include solids, semisolids, liquids, and transdermal therapeutic systems (TTS). Solid compositions are selected from the group consisting of tablets, coated tablets, powders, granules, pellets, capsules, effervescent tablets, or transdermal therapeutic systems. Liquid compositions are also included, selected from the group consisting of solutions, syrups, infusions, extracts, solutions for intravenous application, solutions for infusion, or solutions of the carrier systems of the present invention. Semisolid compositions that can be used in the context of the present invention include emulsions, suspensions, creams, lotions, gels, globules, buccal tablets, and suppositories.

[0116] "Pharmaceutically acceptable" means approved by a regulatory agency of a federal or state government or listed in the United States Pharmacopeia or other generally recognized pharmacopeia for use in animals, especially humans.

[0117] As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic agent is administered. Such pharmaceutical carriers can be sterile liquids, such as saline solutions in water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Saline is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline, aqueous dextrose, and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. If desired, the composition can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by EW Martin.

[0118] The term "fibroblast activation protein (FAP)" as used herein is also known as "seprase." Both terms can be used interchangeably herein. Fibroblast activation protein is a homodimeric integral protein with a dipeptidyl peptidase IV (DPPIV)-like fold, characterized by an α / β hydrolase domain and an eight-bladed β propeller domain.

[0119] Implementation In the following, the various aspects of the invention are defined in more detail. Each aspect so defined may be combined with other aspects, unless expressly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature indicated as being preferred or advantageous.

[0120] In a first aspect, the present invention provides a compound of formula (I): [ka] wherein Q, R, U, V, W, Y, and Z are independently present or absent, provided that at least three of Q, R, U, V, W, Y, and Z are present; Q, R, U, V, W, Y and Z are independently O, CH2, NR 4 , C=O, C=S, C=NR 4 , HCR 4 and R 4 CR 4 wherein no two O's are immediately adjacent to each other; preferably, four of the six groups are present, where two are C=O, one is CH2, and one is NH; more preferably, four groups are present, where two are C=O, one is CH2, and one is NH; most preferably, V, W, Y, and Z are present, where V and Z are C=O, and W and Y are independently selected from CH2 and NH; R 1 and R 2 are independently -H, -OH, halo, C 1-6 -Alkyl, -OC 1-6 -Alkyl and SC 1-6 - selected from the group consisting of alkyl;

[0121] R 3 is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)-alkyl, -C(O)-aryl, -C=CC(O)-aryl, -C=CS(O)2-aryl, -CO2H, -SO3H, -SON2NH2, -PO3H2 and 5-tetrazolyl; R 4 -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each -C1-6 -alkyl is optionally substituted by 1 to 3 substituents selected from -OH, oxo, halo, and may be attached to Q, R, U, V, W, Y, or Z; R 5 -H, halo and C 1-6 - selected from the group consisting of alkyl; R 6 and R 7 independently -H, [ka] where R 6 and R 7 is not simultaneously H; preferably, R 6 is attached to the 7- or 8-quinolyl position, and R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 6 is attached to the 7-quinolyl position, and R 7 is attached to the 6-quinolyl position; where L is a linker; D, A, E and B are independently present or absent, preferably at least A, E and B are present, where, if present: D is a linker;

[0122] A is NR 4 , O, S, and CH2; E is C 1-6 -alkyl, [ka] selected from the group consisting of: where i is 1, 2, or 3; where j is 1, 2, or 3; where k is 1, 2, or 3; where m is 1, 2 or 3; More preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; A and E together form a group selected from cycloalkyl, heterocycloalkyl, aryl and heteroaryl, preferably heterocycloalkyl, where A and E may be monocyclic, bicyclic and polycyclic, preferably monocyclic; each A and E may be -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each of which is optionally substituted by 1 to 4 substituents selected from -C 1-6 -Alkyl may be substituted by 1 to 3 substituents selected from -OH, oxo and halo; and A, B, D, E or [ka] may be bonded to;

[0123] B, S, NR 4 , N.R. 4 -O, NR 4 -C 1-6 -Alkyl, NR 4 -C 1-6 -Alkyl-NR 4 and 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, the heterocycles preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably NR 4 -C 1-6 -Alkyl-NR 4 and the N-containing heterocycle is C 1-6 -Alkyl, aryl, C 1-6 -substituted by 1 to 3 substituents selected from the group consisting of -aralkyl; and R 8 is selected from the group consisting of a radioactive moiety, a chelating agent, a fluorescent dye, an imaging agent, and combinations thereof; [ka] is a 1-naphthyl moiety or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle has two ring atoms between the N atom and X; and the heterocycle may further contain one, two, or three heteroatoms selected from O, N, and S; and X is a C atom. or a pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof. In a preferred embodiment, A and E together form a group selected from C3, C4, C5, C6, C7 and C8 monocyclic heterocycloalkyl, preferably C5 or C6 monocyclic, or C7, C8, C9, C 10 C 11 Or C 12 bicyclic heterocycloalkyl, preferably C7, C8, C9 and C 10 bicyclic heterocycloalkyl, wherein the heterocycloalkyl contains 1, 2, 3 or 4, preferably 1 or 2 heteroatoms independently selected from the group consisting of N, O and S, preferably N and O, and most preferably 1 or 2 N.

[0124] In a preferred embodiment of the first aspect of the present invention, a compound of formula (I): [ka] wherein Q, R, U, V, W, Y, and Z are independently present or absent, provided that at least three of Q, R, U, V, W, Y, and Z are present; Q, R, U, V, W, Y and Z are independently O, CH2, NR 4 , C=O, C=S, C=NR 4 , HCR 4 and R 4 CR 4with the proviso that no two O's are immediately adjacent to each other; preferably, four of the six groups are present, where two are C=O, one is CH2, and one is NH; more preferably, four groups are present, where two are C=O, one is CH2, and one is NH; most preferably, V, W, Y, and Z are present, where V and Z are C=O, and W and Y are independently selected from CH2 and NH; R 1 and R 2 are independently -H, -OH, halo, C 1-6 -Alkyl, -OC 1-6 -Alkyl and SC 1-6 - selected from the group consisting of alkyl;

[0125] R 3 is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)-alkyl, -C(O)-aryl, -C=CC(O)-aryl, -C=CS(O)2-aryl, -CO2H, -SO3H, -SON2NH2, -PO3H2 and 5-tetrazolyl; R 4 -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each -C 1-6 -alkyl is optionally substituted by 1 to 3 substituents selected from -OH, oxo, halo, and may be attached to Q, R, U, V, W, Y, or Z; R 5 -H, halo and C 1-6 - selected from the group consisting of alkyl; R 6 and R 7 independently -H, [ka] where R6 and R 7 is not simultaneously H, preferably, R 6 is attached to the 7- or 8-quinolyl position, and R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 6 is attached to the 7-quinolyl position, and R 7 is attached to the 6-quinolyl position; where L is a linker; D, A, E and B are independently present or absent, preferably at least A, E and B are present, where, if present: D is a linker;

[0126] A is NR 4 , O, S, and CH2; E is C 1-6 -alkyl, [ka] selected from the group consisting of: where i is 1, 2, or 3; where j is 1, 2, or 3; where k is 1, 2, or 3; where m is 1, 2 or 3; More preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl;

[0127] B, S, NR 4 , N.R. 4 -O, NR 4 -C 1-6 -Alkyl, NR 4 -C 1-6 -Alkyl-NR 4 and 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, the heterocycles preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably NR 4 -C1-6 -Alkyl-NR 4 and the N-containing heterocycle is C 1-6 -Alkyl, aryl, C 1-6 -substituted by 1 to 3 substituents selected from the group consisting of -aralkyl; and R 8 is selected from the group consisting of a radioactive moiety, a chelating agent, a fluorescent dye, an imaging agent, and combinations thereof; [ka] is a 1-naphthyl moiety or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle has two ring atoms between the N atom and X; and the heterocycle may further contain one, two, or three heteroatoms selected from O, N, and S; and X is a C atom. or a pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof. Preferably, 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0128] In a further preferred embodiment of the first aspect of the invention, A and E together form a group consisting of C3, C4, C5, C6, C7 and C8 monocyclic heterocycloalkyl, preferably C5 or C6 monocyclic, or C7, C8, C9, C 10 , C 11 Or C 12 Bicyclic heterocycloalkyl groups are formed, preferably C7, C8, C9 and C 10The group comprises a bicyclic heterocycloalkyl, wherein the heterocycloalkyl contains 1, 2, 3 or 4, preferably 1 or 2 heteroatoms independently selected from N, O and S, preferably N and O, and most preferably 1 or 2 N. Preferred monocyclic heterocycloalkyls are selected from the group consisting of pyrrolidinyl, piperidinyl, imidazolidinyl, 1,2-diazacyclohexanyl, 1,3-diazacyclohexanyl, piperazinyl, 1-oxo-2-azacyclohexanyl, 1-oxo-3-azacyclohexanyl or morpholinyl, preferably piperidinyl, piperazinyl and pyrrolidinyl. Preferred bicyclic heterocycloalkyls are selected from the group consisting of bicyclo[2.2.1]2,5-diazaheptanyl, 3,6-diazabicyclo[3.2.1]octanyl, 3,6-diazabicyclo[3.2.2]nonyl, octahydropyrrolo[2,3-b]pyrrolyl, octahydropyrrolo[3,2-b]pyrrolyl, octahydropyrrolo[3,4-b]pyrrolyl, octahydropyrrolo[3,4-c]pyrrolyl, and 9-methyl-3,7,9-triazabicyclo[3.3.1]nonanyl.

[0129] A and E and B on the one hand, and / or R on the other hand 6 Or R 7 The bond between the heterocycles formed by is preferably formed via a heteroatom, preferably N.

[0130] Particularly, preferred examples of the heterocycle formed by A and E include: [ka] is selected from.

[0131] In a preferred embodiment of the first aspect of the present invention, Q, R, and U are CH2 and are independently present or absent; preferably, Q and R are absent; V is CH2, C=O, C=S or C=NR 4 and preferably, V is C=O; W is NR 4 and preferably W is NH; Y is HCR 4 preferably, Y is CH; and Z is C=O, C=S or C=NR 4 and preferably Z is C=O.

[0132] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is CH2; W is NH; Y is CH; and Z is C=O.

[0133] In a further preferred embodiment of the first aspect of the present invention, R 1 and R 2 is independently selected from the group consisting of —H and halo; preferably, R 1 and R 2 is halo; more preferably, R 1 and R 2 is F; R 3 is selected from the group consisting of -H, -CN and -B(OH)2; preferably, R 3 is —CN or —B(OH)2; more preferably, R 3 is -CN; R 4 -H and -C 1-6 -alkyl, wherein -C 1-6 -Alkyl may be substituted by 1 to 3 substituents selected from -OH. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0134] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is CH2; W is NH; Y is CH2; Z is C=O; R 1 and R 2 is independently selected from the group consisting of —H and halo; preferably, R 1 and R 2 is halo; more preferably, R 1 and R 2 is F; R 3 is selected from the group consisting of -H, -CN and -B(OH)2; preferably, R 3 is —CN or —B(OH)2; more preferably, R 3 is -CN; R 4 -H and -C 1-6 -alkyl, wherein -C 1-6 -Alkyl may be substituted by 1 to 3 substituents selected from -OH. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0135] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is CH2; W is CH2; Y is NH; Z is C=O; R 1 and R 2 is independently selected from the group consisting of —H and halo; preferably, R 1 and R 2 is halo; more preferably, R 1 and R 2 is F; R 3 is selected from the group consisting of -H, -CN and -B(OH)2; preferably, R 3is —CN or —B(OH)2; more preferably, R 3 is -CN; R 4 -H and -C 1-6 -alkyl, wherein -C 1-6 -Alkyl may be substituted by 1 to 3 substituents selected from -OH. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0136] In a further preferred embodiment of the first aspect of the present invention, [ka] teeth, [ka] which may contain 1 or 2 heteroatoms selected from O, N and S.

[0137] In a further preferred embodiment of the first aspect of the invention, [ka] teeth, [ka] which may contain 1 or 2 heteroatoms selected from O, N and S.

[0138] In a further preferred embodiment of the first aspect of the invention, [ka] teeth, [ka] R 6 and R7 independently -H, [ka] where R 6 and R 7 is not simultaneously H; preferably, R 6 and R 7 binds to the 5, 6 or 7 position.

[0139] In a preferred embodiment, [ka] teeth, [ka] is selected from the group consisting of: In another preferred embodiment, [ka] teeth, [ka] is.

[0140] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is O, S, CH2, NH or NCH3; E is C 1-6 -Alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; or A and E together [ka] forming a group selected from: B is NR 4 -C 1-6 -Alkyl-NR 4 or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0141] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth, [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is O; E is C 1-6 -Alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -Alkyl-NR 4 or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, preferably further containing 1 or 2 heteroatoms selected from O, N and S, and preferably further containing 1 or 2 nitrogen atoms, wherein the N-containing heterocycle is preferably selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0142] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is S; E is C 1-6 -alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -Alkyl-NR 4 or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0143] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is CH2; E is C 1-6 -Alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0144] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is NH; E is C 1-6 -Alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0145] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is an amino acid, preferably an amino acid with a charged side chain; A is O; E is C 1-6 -alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0146] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is an amino acid, preferably an amino acid with a charged side chain; A is S; E is C 1-6 -alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0147] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is an amino acid, preferably an amino acid with a charged side chain; A is CH2; E is C 1-6 -Alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0148] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is an amino acid, preferably an amino acid with a charged side chain; A is NH; E is C 1-6 -alkyl or [ka] where m is 1, 2 or 3; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is selected from C 1-6 -Alkyl, aryl, C 1-6 -aralkyl. Preferably, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0149] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] Preferably, R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is O; E is C 1-6 -alkyl or [ka] wherein m is 1, 2 or 3; preferably, E is C1-6 -alkyl, and the C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, which heterocycle preferably further contains one or two nitrogen atoms.

[0150] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 5- or 6-quinolyl position; more preferably, R 7 is attached to the 6-quinolyl position, where D is absent; A is O; E is C3 or C4 alkyl; more preferably, E is propyl or butyl; B is a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, which heterocycle preferably further contains one or two nitrogen atoms.

[0151] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is represented by the formula: [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may contain one nitrogen; [ka] is attached to position 1, 2 or 3, preferably to position 2; l is 1 or 2] The aromatic or non-aromatic monocyclic heterocycle is represented by the formula:

[0152] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is represented by the formula: [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may further contain one nitrogen; [ka] is attached to position 1, 2 or 3, preferably to position 2; where the N-containing heterocycle is C 1-6 -substituted by alkyl] The aromatic or non-aromatic monocyclic heterocycle is represented by the formula:

[0153] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is [ka] is selected from the group consisting of where the N-containing heterocycle is C 1-6 -substituted by alkyl, Here, the N-containing heterocycle contained in B is [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, may further contain one nitrogen, and may contain one or more side chains (e.g., derived from amino acids); [ka] is attached to position 1, 2 or 3, preferably to position 2; o is 1 or 2;

[0154] Preferably, the N-containing heterocycle contained in B is [ka] When B is, the N-containing heterocycle contained in B is [ka] more preferably, the N-containing heterocycle contained in B is selected from the group consisting of [ka] When B is [ka] is.

[0155] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is [ka] is selected from the group consisting of Here, the N-containing heterocycle contained in B is [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, may further contain one nitrogen, and may contain one or more side chains (e.g., derived from amino acids);

[0156] [ka] is attached to position 1, 2 or 3, preferably to position 2; o is 1 or 2; Preferably, the N-containing heterocycle contained in B is [ka] When B is, the N-containing heterocycle contained in B is [ka] more preferably, the N-containing heterocycle contained in B is selected from the group consisting of [ka] When B is [ka] is.

[0157] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is [ka] is selected from the group consisting of:

[0158] In a further preferred embodiment of the first aspect of the present invention, The N-containing heterocycle contained in B is [ka] wherein B is selected from the group consisting of C 1-3 It is substituted by alkyl.

[0159] In a further preferred embodiment of the first aspect of the present invention, R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 6-quinolyl position, where D is absent; A is O; E is propyl or butyl; B is [ka] is.

[0160] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is C=O; W is NH; Y is CH2; Z is C=O; R 1 and R 2 is independently selected from the group consisting of —H and halo; preferably, R 1 and R 2 is independently selected from the group consisting of —H and F; more preferably, R 1 and R 2 are identical and selected from the group consisting of -H and F; R 3 is -CN; R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 6-quinolyl position, where D is absent;

[0161] A is O; E is C 1-6 -alkyl or [ka] wherein m is 1, 2 or 3; preferably, E is C 1-6 -alkyl; preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; more preferably, E is C 1-6 -alkyl, most preferably E is C3 or C4 alkyl; B is NH-C 1-6 -alkyl, [ka] and preferably C 1-6 -alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl; preferably, B is [ka] and [ka] teeth [ka] is.

[0162] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is C=O; W is NH; Y is CH2; Z is C=O; R 1 and R 2 are identical and selected from the group consisting of -H and F; R 3 is -CN; R 5 and R 6 is H; R 7 teeth [ka] and preferably R 7 is attached to the 6-quinolyl position, where D is absent; A is O, S, CH2, NH or NCH3; E is methyl, ethyl, propyl, or butyl;

[0163] A and E together [ka] forming a group selected from: B is [ka] and B is C 1-3 and optionally substituted by alkyl; preferably, B is [ka] and [ka] teeth [ka] is.

[0164] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is C=O; W is NH; Y is CH2; Z is C=O; R 1 and R 2 are identical and selected from the group consisting of -H and F; R 3 is -CN; R 5 and R 6is H; R 7 teeth [ka] and preferably R 7 is attached to the 6-quinolyl position, where D is absent; A is O; E is methyl, ethyl, propyl, or butyl; B is [ka] and preferably B is [ka] and [ka] teeth [ka] is.

[0165] In a further preferred embodiment of the first aspect of the present invention, Q, R and U are absent; V is C=O; W is NH; Y is CH2; Z is C=O; R 1 and R 2 are identical and selected from the group consisting of -H and F; R 3 is CN; R 5 and R 6 is H; R 7 teeth [ka] and R 7is attached to the 6-quinolyl position, where D is absent; A is O; E is methyl, ethyl, propyl, or butyl; B is [ka] Preferably, B is [ka] and [ka] teeth [ka] is.

[0166] In a first further preferred embodiment of the present invention, C 1-6 -Alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl.

[0167] In a first further preferred embodiment of the present invention, C 1-3 -Alkyl is selected from the group consisting of methyl, ethyl, propyl and i-propyl.

[0168] In a first further preferred embodiment of the present invention, C 1-6 -aralkyl is selected from the group consisting of benzyl, phenyl-ethyl, phenyl-propyl and phenyl-butyl.

[0169] In further preferred embodiments of the first aspect of the invention, the compound of the first aspect of the invention is selected from the compounds of Table 1. More preferably, the compound of the first aspect of the invention is selected from the compounds of Table 2. More preferably, the compound of the first aspect of the invention is selected from the group consisting of FAPI-02 and FAPI-04.

[0170] In a preferred embodiment of the first aspect of the present invention, the compound of the first aspect of the present invention is selected from the compounds in Table 1 and / or Table 3. More preferably, the compound of the first aspect of the present invention is selected from the compounds in Table 2 and / or Table 4. More preferably, the compound of the first aspect of the present invention is selected from the group consisting of FAPI-02, FAPI-04, FAPI-46, FAPI-34, FAPI-42, FAPI-52, FAPI-69, FAPI-70, FAPI-71, FAPI-72 and FAPI-73.

[0171] Table 1: Preferred compounds of the first aspect of the invention §Fluorescent compounds;$ 99m Tc-chelating agent; *Pb-chelating agent; R 1 and R 2 is at the 4-pyrrolidine position; Q, R and U are absent; [ka] teeth [ka] and R 5 is H;R 6 is attached to the 7-quinolyl position; R 7 is attached to the 6-quinolyl position; "-" is R 6 or R 7 indicates H; "+" indicates R 6 or R 7 but [ka] V is C=O; W is NH; Y is CH; Z is C=O; R 3 is -CN; A is O (A is absent and R 7 binds to the 5-quinolyl position (FAPI-01 excluded). [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0172] Table 2: Compounds of particular interest. Q, R, U and D are absent; R 1 and R 2 is at the 4-pyrrolidine position; [ka] teeth [ka] and R 5 , R 6 is H;R 7 is attached to the 6-quinolyl position; V is C=O; W is NH; Y is CH; Z is C=O; R 3 is -CN; B is 1,4-piperazine; E is 1,3-propane; and A is O. [Table 2]

[0173] Table 3: Further preferred compounds according to the first aspect of the invention. §Fluorescent compounds;$ 99m Tc-chelating agent;* 18 F-precursor of the label; Q, R and U are absent; R 1 and R 2 is at the 4-pyrrolidine position; [ka] teeth [ka] and R5 and R 6 is H;R 7 is attached to the 6-quinolyl position, [ka] V is C=O; W is NH; Y is CH; Z is C=O; R 3 is -CN. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0174] Table 4: Compounds of particular interest. Q, R, U and D are absent; R 1 and R 2 is a fluorine atom at the 4-pyrrolidine position; [ka] teeth [ka] and;R 5 , R 6 is H;R 7 is attached to the 6-quinolyl position; V is C=O; W is NH; Y is CH; Z is C=O; R 3 is -CN; B is 1,4-piperazine; E is 1,3-propane; and A is O. [Table 4]

[0175] [Table 5-1] [Table 5-2]

[0176] In a further preferred embodiment of the first aspect of the present invention, R 8 is a radioactive moiety that is a fluorescent isotope, a radioisotope, a radiopharmaceutical, or a combination thereof. Preferably, the radioactive moiety is an α-radiation emitting isotope, a β-radiation emitting isotope, a γ-radiation emitting isotope, an Auger electron emitting isotope, an X-ray emitting isotope, a fluorescent emitting isotope, e.g. 11 C. 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 212 Pb, 64 Cu,67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Preferably, the compound is selected from the group consisting of: 18 F, 64 Cu, 68 Ga, 90 Y, 99m Tc, 153 Sm, 177 Lu, 188 Re and the like.

[0177] In a further preferred embodiment of the first aspect of the present invention, R 8 are fluorescent dyes selected from the group consisting of the following types of fluorescent dyes: xanthenes, acridines, oxazines, cyanines, styryl dyes, coumarins, porphines, metal-ligand complexes, fluorescent proteins, nanocrystals, perylenes, boron-dipyrromethenes, and phthalocyanines, as well as composites and combinations of these types of dyes.

[0178] In a further preferred embodiment of the first aspect of the present invention, R 8 is a chelating agent that forms a complex with divalent or trivalent metal cations. Preferably, the chelating agent is selected from the group consisting of 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), triethylenetetramine (TETA), iminodiacetic acid, diethylenetriamine-N,N,N',N',N''-pentaacetic acid (DTPA), bis-(carboxymethylimidazole)glycine, and 6-diethylazinopyridine-3-carboxylic acid (HYNIC).

[0179] In a first further preferred embodiment of the present invention, R 8 is a contrast agent comprising or consisting of a paramagnetic agent, preferably wherein the paramagnetic agent comprises or consists of paramagnetic nanoparticles.

[0180] In a first further preferred embodiment of the present invention, R 8 is any of the R in Tables 1 to 5 8 is.

[0181] In a second aspect, the present invention relates to a pharmaceutical composition comprising or consisting of at least one compound of the first aspect and a pharmaceutically acceptable carrier and / or excipient.

[0182] In a third aspect, the present invention relates to a compound of the first aspect or a pharmaceutical composition of the second aspect for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in an animal or human subject. Preferably, the disease characterized by overexpression of fibroblast activation protein (FAP) is selected from the group consisting of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling, and keloid disorders.

[0183] Preferably, when the disease characterized by overexpression of fibroblast activation protein (FAP) is cancer, said cancer is selected from the group consisting of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal carcinoma, laryngeal cancer, myeloma cell, bladder cancer, cholangiocarcinoma, renal clear cell carcinoma, neuroendocrine tumor, tumor-induced osteomalacia, sarcoma, CUP (cancer of unknown primary stage), thymic carcinoma, desmoid tumor, glioma, astrocytoma, cervical cancer and prostate cancer.Preferably, said cancer is glioma, breast cancer, colon cancer, lung cancer, head and neck cancer, liver cancer or pancreatic cancer.More preferably, said cancer is glioma.

[0184] Preferably, when the disease characterized by overexpression of fibroblast activation protein (FAP) is chronic inflammation, the chronic inflammation is selected from the group consisting of rheumatoid arthritis, osteoarthritis, and Crohn's disease. Preferably, the chronic inflammation is rheumatoid arthritis.

[0185] Preferably, when the disease characterized by overexpression of fibroblast activation protein (FAP) is a fibrosis, said fibrosis is selected from the group consisting of pulmonary fibrosis, such as idiopathic pulmonary fibrosis, and liver cirrhosis.

[0186] Preferably, when the disease characterized by overexpression of fibroblast activation protein (FAP) is tissue remodeling, said tissue remodeling occurs after myocardial infarction.

[0187] Preferably, when the disease characterized by overexpression of fibroblast activation protein (FAP) is a keloid lesion, said keloid lesion is selected from the group consisting of scar formation, keloid tumors and keloid scars.

[0188] In a fourth aspect, the present invention relates to a kit comprising or consisting of a compound of the first aspect or a pharmaceutical composition of the second aspect and instructions for the diagnosis or treatment of a disease, preferably the disease being one of the specific diseases described above. [Example]

[0189] Example 1: Compound synthesis and radiochemistry Two radiotracers were synthesized based on the FAP-α specific inhibitor (Jansen et al., ACS Med Chem Lett, 2013). Radioiodinated FAPI-01 was obtained via an organotin stannylated precursor prepared by palladium-catalyzed bromide / tin exchange. FAPI-02 is a precursor for a radiometal chelate synthesized in five steps. Additional compounds were prepared by applying the same or slightly modified procedures. The structures of these compounds are listed in Tables 1 and 2. Radioiodination of the stannylated precursor was carried out using peracetic acid. For chelation with Lu-177 and Ga-68, the pH of the reaction mixture was adjusted with sodium acetate and heated to 95°C for 10 min. Stability in human serum was analyzed by radio-HPLC analysis of the precipitation and supernatant.

[0190] reagent All solvents and non-radioactive reagents were obtained in reagent grade from ABCR (Karlsruhe, Germany), Sigma-Aldrich (Muenchen, Germany), Acros Organics (Geel, Belgium), or VWR (Bruchsal, Germany) and used without further purification. Atto 488 NHS-ester was obtained from AttoTec (Siegen, Germany). 2,2',2''-(10-(2-(4-nitrophenyl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTA-PNP) was synthesized according to the protocol of Mier et al. (Mier et al., Bioconjug Chem, 2005). The intermediates 6-methoxyquinoline-4-carboxylic acid (7), 5-bromoquinoline-4-carboxylic acid (3), and (S)-1-(2-aminoacetyl)pyrrolidine-2-carbonitrile methyl 4-benzenesulfonate were synthesized according to the protocol described by Jansen et al. (Jansen et al., ACS Med Chem Lett, 2013). The compound (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-5-bromoquinolinecarboxamide was synthesized by a modified HBTU amidation protocol.

[0191] Compound synthesis Scheme 1 shows the initial synthesis of FAPI-01, which involves Br / Li exchange with n-butyllithium on 5-bromoquinolyl-4-carboxylic acid (3) and quenching with elemental iodine to give iodoquinoline 4. This compound was coupled to the Gly-Pro-CN fragment via HBTU / HOBt activation to give the non-radioactive counterpart of FAPI-01 (1). [ka] Scheme 1. Synthesis of non-radioactive FAPI-01. i) nBuLi, then I2, THF; ii) HBTU / HOBt, DIPEA, H-Gly-Pro-CN, DMF.

[0192] In the synthesis of radioactive FAPI-01 (1*), palladium-catalyzed stannylation of inhibitor 5 in dioxane at 80° C. gave the stannylated precursor 6 (Scheme 2). [ka] Scheme 2. Synthesis of radioactive FAPI-1 via stannylated precursor 4. i) (Me3Sn)2; (PPh3)2PdCl2; dioxane 80 °C; ii) I-125 or I-131; AcOOH; 1 M HCl; MeOH.

[0193] To enable radiolabeling via incorporation of radioactive metals, the chelator DOTA was chemically attached to the basic scaffold of the FAP inhibitor. As demonstrated by Jansen et al. (ACS Med Chem Lett, 2013), modification at the 6-position of quinoline-4-carboxylic acid is well tolerated without compromising target affinity and specificity. Therefore, a bifunctional linker was attached to the hydroxyl group of 8 via an ether bond, which became the primary method for the synthesis shown in Scheme 3. Readily available 1-bromo-3-chloropropane was selected to create a spacer that remained intact during saponification of the simultaneously formed ester bond at the end of the one-pot process. Compound 9 was converted to N-Boc-protected quinolinic carboxylic acid 10, which was further coupled to H-Gly-Pro-CN with HBTU. Because the free amine is highly hygroscopic, compound 11 was directly converted to FAPI-02 (2) after Boc removal, solvent exchange, and neutralization of excess p-toluenesulfonic acid. [ka] Scheme 3. Chemical synthesis of FAPI-02. i) q HBr 48%, 130 °C; ii) 1-bromo-3-chloropropane, Cs2CO3, DMF, then 6M NaOH; iii) 1-Boc-piperazine, KI, DMF; iv) HBTU / HOBt, DIPEA, H-Gly-Pro-CN, DMF; v) TosOH, MeCN, then DOTA-PNP, DIPEA, DMF.

[0194] For compounds incorporating the group A≠O, the quinoline-4-carboxylic acid intermediate was synthesized via an alternative reaction scheme. The key step in this approach is a palladium-catalyzed coupling reaction (e.g., Buchwald-Hartwig cross-coupling), which requires additional protection prior to the cross-coupling reaction and deprotection of the carboxylic acid function after the cross-coupling reaction (Scheme 4). [ka] Scheme 4. Synthesis of 6-(3-(4-Boc-piperazin-1-yl)propyl-1-(methyl)amino)quinoline-4-carboxylic acid, a building block for the synthesis of FAPI-46. i) DCC, tBuOH, CuCl; ii) 3-methylamino-1-propanol, CsCO, Pd(dba), BINAP; iii) MsCl, NEt, DCM followed by 1-Boc-piperazine, KI, DMF; iv) TFA followed by BocO, NEt, DMF.

[0195] [ka] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-5-trimethylstannylquinolinecarboxamide (6) 3.88 mg (10.0 μmol) of (S)—N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-5-bromoquinolinecarboxamide, 20 μL (32 mg; 96 μmol) of hexamethylditin, and 0.75 mg (1.07 μmol) of bis(triphenylphosphine)palladium(II) dichloride in 1 mL of dry dioxane are stirred overnight at 80 °C under an inert atmosphere. The volatiles are removed, and the residue is taken up in 2 mL of 50% acetonitrile / water, filtered through a C18 light cartridge, and then purified by HPLC. After lyophilization, 2.78 mg (5.90 μmol; 59%) of the product is obtained. LC-MS R t 14.77 min, m / z 473.0786 [M( 120 Sn)+H] +

[0196] [ka] 5-Iodoquinol-4-carboxylic acid (4) 5.42 mg (136 μmol) of sodium hydride suspension (60% in mineral oil) was added to a solution of 30.27 mg (120 μmol) of 5-bromoquinoline-4-carboxylic acid (3) in 3 mL of dry THF at 0 °C under Ar. The ice bath was removed, and the reaction mixture was cooled to -78 °C. 100 μL (160 μmol) of nBuLi (1.6 M in hexane) was then added dropwise. After 15 min, 64.71 mg (254 μmol) of iodine in 2 mL of THF was added dropwise, and the reaction was stirred at -78 °C for 30 min before allowing to reach room temperature. After 1 h, the reaction was quenched by adding 1 mL of 0.5 M NaHCO3 and approximately 30 mg (170 μmol) of sodium dithionite to remove excess iodine. After removing the THF under reduced pressure, the mixture was acidified to pH 2 and extracted three times with ethyl acetate (25 mL). The organic phases are combined, evaporated to dryness and purified by HPLC to give, after lyophilization, 18.14 mg (60.7 μmol; 45%) of the title compound. 1H NMR (500 MHz, DMSO-d6) 13.95 (br, 0.3H), 8.93 (s, 1H), 8.34 (d, J =7.2 Hz, 1H), 8.12 (d, J = 8.4 Hz, 1H), 7.60 (s, 1H), 7.52 (t, J = 7.9 Hz, 1H); 13 C NMR (125 MHz, DMSO-d6) 168.8, 150.3, 148.8, 141.3, 130.6, 121.0, 109.5; LC-MS R t 8.65 min, m / z 299.9383 [M+H] +

[0197] [ka] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-5-trimethylstannylquinolinecarboxamide (1; FAPI-01) 9.07 mg (23.9 μmol) of HBTU in 50 μL of DMF is added to a solution of 6.21 mg (20.8 μmol) of 5-iodoquinoline-4-carboxylic acid, 7.45 mg (55.2 μmol) of HOBt, and 10 μL of DIPEA in 50 μL of DMF. After 15 minutes, (S)-1-(2-aminoacetyl)pyrrolidine-2-carbonitrile methyl 4-benzenesulfonate (29.9 μmol) in 50 μL of DMF is added. The reaction is quenched with 850 μL of water and purified by HPLC. Lyophilization yields 6.86 mg (15.8 μmol; 76%) of product. 1 H NMR (600 MHz, DMSO-d6) 9.06, 8.97, 8.33, 8.13, 7.56, 7.51, 4.81, 4.34, 4.06, 3.74, 3.56, 2.21, 2.17, 2.09, 2.05; 13C NMR (150 MHz, DMSO-d6) 167.1, 150.2, 148.8, 145.3, 141.5, 130.7, 125.3, 121.9, 119.3, 92.0, 46.3, 45.4, 42.1, 29.5, 24.9; LC-MS R t 11.95 min, m / z 435.0102 [M+H] +

[0198] [ka] 6-Hydroxyquinoline-4-carboxylic acid (8) 105 mg (477 μmol) of crude 6-methoxyquinoline-4-carboxylic acid (7) is dissolved in 3 mL of 48% aqueous hydrobromic acid. The solution is heated to 130°C for 4 hours. After the solution reaches room temperature, it is brought to a slightly basic pH with 6 M NaOH. After HPLC purification and lyophilization, 79.2 mg (419 μmol; 88%) of product is obtained. 1 H NMR (500 MHz, DMSO-d6) 13.65 (br, 0.6H) 10.24 (s, 1H), 8.78 (d, J = 4.4 Hz, 1H), 8.06 (d, J = 2.6 Hz, 1H), 7.95 (d, J = 9.1 Hz, 1H), 7.84 (d, J = 4.4 Hz, 1H), 7.37 (dd, J = 9.1, 2.6 Hz, 1H), 13 LC-MS R t 6.66 min, m / z 190.0415 [M+H] +

[0199] [ka] 6-Bromoquinoline-4-carboxylic acid tert-butyl ester 98.3 mg (390 μmol) of 6-bromoquinoline-4-carboxylic acid (neat) was suspended in 5 mL of tetrahydrofuran and 25.0 μL (18.3 mg; 181 μmol) of triethylamine and added to O-tert-butyl-N,N'-dicyclohexylurea (prepared the previous day from 426 mg (2.07 mmol) of pure dicyclohexylcarbodiimide, 173 mg (2.33 mmol) of pure tert-butanol, and 10.2 mg (103 μmol) of pure copper(I) chloride). The mixture was heated to 50 °C overnight. The mixture was filtered, the solvent evaporated, and the product isolated by HPLC. After lyophilization, 49.7 mg (161 μmol; 41%) of the title compound was obtained. LC-MS R t 20.40 min, m / z 251.9642 [M-tBu] +

[0200] [ka] 6-(3-chloro-1-propoxy)quinoline-4-carboxylic acid (9) 42.4 μL (67.4 mg; 430 μmol) of 1-bromo-1-chloropropane was added to a suspension of 23.2 mg (123 μmol) of 6-hydroxyquinoline-4-carboxylic acid (8) and 190 mg (1.38 μmol) of potassium carbonate in 250 μL of DMF and heated to 60 °C overnight. The reaction mixture was cooled to room temperature and diluted with 500 μL of water and 500 μL of acetonitrile, followed by the addition of 100 μL of 6 M NaOH. The reaction mixture was directly purified by HPLC (5-40%) after complete ester hydrolysis. After lyophilization, 26.45 mg (99.4 μmol; 81%) of product was obtained. 1H NMR (500 MHz, DMSO-d6) 13.75 (br, 0.4H), 8.88 (d, J = 4.4 Hz, 1H), 8.19 (d, J = 2.0 Hz, 1H), 8.04 (d, J = 9.2 Hz, 1H), 7.94 (d, J = 4.4 Hz, 1H), 7.52 (dd, J = 9.2, 2.0 Hz, 1H), 4.24 (t, J = 5.95 Hz, 2H), 3.85 (t, J = 6.5 Hz, 2H), 2.27 (m, 2H); 13 LC-MS R t 11.46 min, m / z 266.0461 [M+H] +

[0201] [ka] tert-Butyl 6-(3-hydroxypropylmethylamino)quinoline-4-carboxylate 204.6 mg (664 μmol) of tert-butyl 6-bromoquinoline-4-carboxylate, 34.10 mg (54.7 μmol) of BINAP, 321.51 mg (23.5 μmol) of Pd2(dba), and 480.3 mg (1.47 mmol) of cesium carbonate were dissolved in 6 mL of toluene, and 128.0 μL (118 mg; 1.32 mmol) of N-methyl-1,3-propanolamine was added. After stirring overnight at 90 °C, the solvent was removed, and the residue was suspended in 1:1 water / acetonitrile, filtered, and purified by HPLC. After lyophilization, 172.7 mg (547 μmol; 82%) of the title compound was obtained. LC-MS R t 13.41 min, m / z 261.1213 [M-tBu+H] +

[0202] [ka] tert-Butyl 6-(3-(4-Boc-piperazin-1-yl)propyl-1-(methyl)amino)quinoline-4-carboxylate 62.8 mg (199 μmol) of tert-butyl 6-(3-hydroxypropylmethylamino)quinoline-4-carboxylate was dissolved in 5 mL of dichloromethane and 90.0 μL (66.6 mg; 659 μmol) of triethylamine. 20.0 μL (29.6 mg; 258 μmol) of methanesulfonyl chloride was added at 0°C, and the mixture was reacted for 60 minutes. 194.6 mg (1.05 mmol) of 1-Boc-piperazine was added, and the volatiles were removed. 500 μL of dimethylformamide and 47.4 mg (286 μmol) of potassium iodide were added to the residue. The mixture was shaken at 60°C for 120 minutes, and the product was isolated by HPLC. After lyophilization, 81.05 mg (167 μmol; 84%) of the title compound was obtained. LC-MS R t 13.99 min, m / z 485.3086 [M+H] +

[0203] [ka] 6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxylic acid (10) 15.13 mg (56.9 μmol) of 6-(3-chloro-1-propoxy)quinoline-4-carboxylic acid (9), 55.43 mg (298 μmol) of N-tert-butoxycarbonylpiperazine, and 51.05 mg (30.8 μmol) of potassium iodide were dissolved in 250 μL of DMF. The mixture was shaken overnight at 60 °C. The resulting suspension was diluted with 750 μL of water, and the product was purified by HPLC. After lyophilization, 28.73 mg (54.3 μmol; 95%) of the product was obtained as the corresponding TFA salt. 1H NMR (500 MHz, D2O) 8.93 (d, J = 5.5 Hz, 1H), 8.17 (d, J = 9.3 Hz, 1H), 7.94 (d, J = 5.5 Hz, 1H), 7.79 (dd, J = 9.3, 2,5 Hz, 1H), 7.65 (d, J = 2.5 Hz, 1H), 4.36 (t, J = 5.6 Hz, 2H), 4.27 (d, J = 13.55 Hz, 2H), 3.67 (d, J = 11.95 Hz), 3.47 (t, J = 15.5 Hz, 2H), 3.27 (t, J = 12.7 Hz), 3.12 (td, J = 12.2, 2.65 Hz), 2.37 (m2 H), 1.47 (s, 9H); 13 C NMR (125 MHz, D2O) 155.5, 153.5, 149.0, 141.4, 134.4, 127.9, 126.6, 122.3, 118.4, 110.0, 105.1, 82.8, 65.5, 54.3, 51.5, 48.6, 40.7, 29.6, 27.4; LC-MS R t 10.62 min, m / z 416.1997 [M+H] +

[0204] [ka] 6-(3-(4-Boc-piperazin-1-yl)propyl-1-(methyl)amino)quinoline-4-carboxylic acid 100.12 mg (206 μmol) of tert-butyl 6-(3-(4-Boc-piperazin-1-yl)propyl-1-(methyl)amino)quinoline-4-carboxylate was treated with 900 μL of trifluoroacetic acid, 25 μL of triisopropylsilane, 25 μL of water, and 50 μL of trifluoromethylsulfonic acid for 60 min. The deprotected compound was precipitated with diethyl ether, dried, and reacted with 60.83 mg (279 μmol) of di-tert-butyl dicarbonate and 50.0 μL of triethylamine (36.5 mg; 361 μmol) in 1 mL of dimethylformamide for an additional 60 min. After HPLC purification and lyophilization, 55.42 mg (129 μmol; 65% over two steps) of the product was obtained. LC-MS R t 10.52 min, m / z 429.2463 [M+H] +

[0205] [ka] (S)-N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide (11) 9.43 mg (24.9 μmol) of HBTU in 50 μL of DMF is added to a solution of 10.56 mg (19.9 μmol) of 6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxylic acid (10), 5.38 mg (39.8 μmol) of HOBt, and 10 μL of DIPEA in 50 μL of DMF. After 15 minutes, 29.9 μmol of (S)-1-(2-aminoacetyl)pyrrolidine-2-carbonitrile methyl 4-benzenesulfonate in 50 μL of DMF is added. The reaction is quenched with 850 μL of water and purified by HPLC. Lyophilization affords 12.88 mg (19.4 μmol; 97%) of the title compound. 1H NMR (500 MHz, DMSO-d6) 9.04 (d, J = 5.5 Hz, 1H), 8.24 (d, J = 9.6 Hz, 1H), 8.10 (d, J = 5.5 Hz, 1H), 7.89 (d, J = 2.3 Hz, 1H), 7.85 (dd, J = 9.6, 2.3 Hz, 1H), 4.84 (t, J = 6 Hz, 1 H), 4.46-4.36 (m, 4H), 4.26 (d, J = 12.0 Hz, 2H), 3.83 (m, 1H), 3.67 (m, 3H), 3.47 (t, J = 7.7 Hz, 2H), 3.27 (br, 2H), 3.11 (t, J = 11.5 Hz), 2.37 (m, 4H), 2.22 (m, 2H), 1.46 (s, 9H); 13 C NMR (125 MHz, DMSO-d6) 168.6, 168.0, 159.4, 155.5, 147.7, 141.8, 135.1, 128.2, 127.5, 123.1, 120.0, 119.1, 104.7, 82.9, 66.0, 54.3, 51.5, 47.0, 46.3, 42.3, 29.4, 27.4, 24.7, 23.1; LC-MS R t 11.81 min, m / z 551.2736 [M+H] +

[0206] [ka] (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonyl-piperazin-1-yl)-1-propoxy)quinoline-4-carboxamide The previous protocol was followed to obtain 13.2 mg (22.4 μmol; 75%). LC-MS R t 11.84 min, m / z 605.2610 [M+H] +

[0207] [ka] N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-Boc-piperazin-1-yl)propyl-1-(methyl)amino)quinoline-4-carboxamide The previous protocol was followed to obtain 1.17 mg (1.95 μmol; 92%). LC-MS R t 12.66 min, m / z 600.3057 [M+H] +

[0208] [ka] FAPI-02(2) 4.85 mg (8.80 mmol) of (S)—N-(2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonyl-piperazin-1-yl)-1-propoxy)quinoline-4-carboxamide (11) is dissolved in 1 mL of acetonitrile and 4.2 mg (22.0 μmol) of 4-methylbenzenesulfonic acid monohydrate is added. The reaction is shaken overnight at 45° C., and then the volatiles are removed under reduced pressure. The residue is taken up in 190 μL of dimethylformamide and 10 μL of triethylamine (7.3 mg; 72 μmol), and then 6.77 mg (12.9 mmol) of DOTA-p-nitrophenol ester is added. The reaction mixture is diluted with 1 mL of water, shaken for 2 hours, and then purified by HPLC. After lyophilization, 5.04 mg (6.02 μmol; 68%) is obtained. 1 H NMR (600 MHz, D2O) 9.02, 8.23, 8.07, 7.87, 7.83, 4.85, 4.45, 4.41, 4.40, 4.39, 3.83, 3.67, 3.50, 3.49, 2.40, 2.38, 2.36, 2.26, 2.22, 2.16; 13C NMR (150 MHz, D2O) 167.9, 159.1, 147.2, 141.8, 135.4, 127.9, 127.2, 119.8, 119.0, 104.5, 65.8, 54.1, 46.8, 46.1, 42.1, 29.2, 24.5, 23.0: LC-MS R t 8.37 min, m / z 837.3872 [M+H] +

[0209] [ka] FAPI-04 The previous protocol was followed to obtain 3.97 mg (4.55 μmol; 57%). LC-MS R t 8.80 min, m / z 873.3664 [M+H] +

[0210] [ka] FAPI-42 The previous protocol was followed to obtain 1.91 mg (2.47 μmol; 88%). LC-MS R t 9.37 min, m / z 386.6807 [M+2H] 2+

[0211] [ka] FAPI-46 The previous protocol was followed to obtain 39.21 mg (44.3 μmol; 85%). LC-MS R t 9.03 min, m / z 443.7196 [M+2H] 2+

[0212] [ka] FAPI-19 1.09 mg (1.86 μmol) of (S)—N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide was preactivated with 2.13 mg (5.62 μmol) of HBTU and 2.50 μL (1.85 mg; 14.3 μmol) of DIPEA, Boc-deprotected by the method applied to FAPI-02, and reacted with 2.74 mg (5.91 μmol) of bis((1-(2-(tert-butoxy)-2-oxoethyl)-1H-imidazol-2-yl)methyl)glycine. After HPLC purification and solvent removal, the residue was treated with 200 μL of 2.5% trifluoromethanesulfonic acid in 1:1 acetonitrile / trifluoroacetic acid. After precipitation with diethyl ether and HPLC purification, 1.06 mg (1.29 μmol; 70%) of the title compound was obtained. LC-MS R t 8.91 min, m / z 820.2933 [M+H] +

[0213] [ka] FAPI-28 1.00 μL (0.74 mg; 5.73 μmol) of DIPEA was added to a solution of 0.95 mg (1.16 μmol) of FAPI-19, 0.42 mg (3.14 μmol) of HOBt, and 1.10 mg (2.89 μmol) of HBTU in 50 μL of DMF. After 10 min, 2.30 mg (5.34 μmol) of H-Asn(Trt)-OtBu was added and the reaction was allowed to proceed for 120 min. The tert-butyl protecting group was removed with 2.5% TfOH in 8:2 TFA / acetonitrile. After HPLC purification and lyophilization, 0.79 mg (0.75 μmol; 65%) of the title compound was obtained. LC-MS R t 9.23 min, m / z 524.7100 [M+2H] 2+

[0214] [ka] FAPI-34 The previous protocol was followed to obtain 1.01 mg (0.87 μmol; 52%). LC-MS R t 8.87 min, m / z 583.6988 [M+2H] 2+

[0215] [ka] FAPI-60 3.91 mg (6.66 μmol) of (S)—N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide was deprotected with 50 μL of acetonitrile and 100 μL of trifluoroacetic acid for 30 minutes. After evaporation of the solvent and washing with diethyl ether, a mixture of 8.02 mg (9.27 μmol) of acetyl-Cys(Trt)-Gly-Cys(Trt)-Gly-OH, 4.31 mg (31.9 μmol) of HOBt, and 4.47 mg (11.8 μmol) of HBTU in 150 μL of dimethylformamide and 2.50 μL (1.85 mg; 14.3 μmol) of DIPEA was added to the residue and reacted for 120 min. After HPLC purification and lyophilization, 4.66 mg (3.49 μmol; 52%) of the S-trityl-protected title compound was obtained. 3.36 mg (2.52 μmol) of the trityl-protected compound was dissolved in 50 μL of acetonitrile. 3 μL of triethylsilane and 100 μL of trifluoroacetic acid were added and the mixture was allowed to react for 30 minutes. After HPLC purification and lyophilization, 2.01 mg (2.36 μmol; 94%; 49% over two steps) of the title compound was obtained. LC-MS R t 10.26 min, m / z 871.2703 [M+Na] +

[0216] [ka] FAPI-69 The previous protocol was followed to obtain 0.59 mg (0.60 μmol; 39%). LC-MS R t 10.25 min, m / z 991.3490 [M+H] +

[0217] [ka] FAPI-70 The previous protocol was followed to obtain 0.61 mg (0.54 μmol; 33%). LC-MS R t 10.14 min, m / z 1120.3884 [M+H] +

[0218] [ka] FAPI-71 The previous protocol was followed to obtain 0.79 mg (0.66 μmol; 34%). LC-MS R t 10.17 min, m / z 596.7075 [M+2H] 2+

[0219] [ka] Atto488-FAPI-02(14) 0.66 mg (1.20 μmol) of compound 11 was treated with 1.33 mg (6.96 μmol) of 4-methylbenzenesulfonic acid monohydrate in 250 μL of acetonitrile at 45° C. for 4 hours. After removing the solvent, the residue was dissolved in 95 μL of dimethylformamide and 5 μL of triethylamine (3.65 mg; 36.1 μmol). 0.54 mg (0.55 μmol) of Atto 488 NHS-ester in 25 μL of DMSO was added. After 60 minutes, 0.49 mg (0.43 μmol; 78%) of the title compound was isolated by HPLC and lyophilized. LC-MS R t10.19 min, m / z 1022.2706 [M] +

[0220] [ka] FAPI-73 10.95 mg (18.7 μmol) of (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-tert-butoxycarbonylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide was deprotected with 100 μL of acetonitrile and 200 μL of trifluoroacetic acid for 30 min. After evaporation of the solvent and washing with diethyl ether, 15.02 mg (9.27 μmol) of N,N,N-trimethyl-5-((2,3,5,6-tetrafluorophenoxy)-carbonyl)pyridin-2-aminium chloride was added, and the mixture was dissolved in 200 μL of dimethylformamide and 10.0 μL of triethylamine (7.30 mg; 72.3 μmol). After 120 min, the mixture was purified by HPLC and lyophilized to give 11.24 mg (14.7 μmol; 79%) of the title compound. LC-MS R t 9.37 min, m / z 649.2892 [M-CF3CO2] +

[0221] [ka] FAPI-72 The previous protocol was followed to obtain 9.80 mg (12.6 μmol; 70%). LC-MS R t 9.28 min, m / z 662.3237 [M-CF3CO2] +

[0222] General conjugation of side-chain protected Fmoc-amino acids [ka] (S)-N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-(γ,γ-di-tert-butyl)-L-carboxy-glutamylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide 14.04 mg (23.9 μmol) of (S)—N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(1-tert-butoxycarbonyl-piperidin-4-yl)-1-propoxy)quinoline-4-carboxamide was dissolved in 50 μL of acetonitrile and 100 μL of trifluoroacetic acid. After 10 min, the volatiles were removed and the residue was washed with diethyl ether. A solution of 14.95 mg (28.4 μmol) of Fmoc-L-Gla(tBu)2-OH, 7.74 mg (57.4 μmol) of HOBt, 13.46 mg (35.5 μmol) of HBTU, and 20.0 μL (14.8 mg; 115 μmol) of DIPEA in 200 μL of dimethylformamide was added to the dried residue. After 60 minutes, 50.0 μL (50.4 mg; 578 μmol) of morpholine was added, and after 30 minutes the product was isolated by HPLC to give 15.95 mg (20.7 μmol; 86%) of the title compound after lyophilization. LC-MS R t 12.85 min, m / z 772.3643 [M+H] +

[0223] [ka] FAPI-75 3.37 mg (4.37 μmol) of (S)—N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(4-(γ,γ-di-tert-butyl)-L-carboxyglutamylpiperazin-1-yl)-1-propoxy)quinoline-4-carboxamide and 4.52 mg (10.7 μmol) of NOTA-p-nitrophenol were dissolved in 100 μL of dimethylformamide and 10.0 μL of triethylamine (7.30 mg; 72.3 μmol). After HPLC purification and lyophilization, the intermediate compound was deprotected by incubation for 60 min in a solution of 50 μL of acetonitrile, 100 μL of trifluoroacetic acid, 2.5 μL of triisopropylsilane, and 2.5 μL of water. After HPLC-purification and lyophilization, 2.62 mg (2.77 μmol; 63%) was obtained. LC-MS R t 9.38 min, m / z 945.3668 [M+H] +

[0224] [ka] FAPI-77-precursor The general active ester modification protocol was followed to obtain 3.23 mg (3.06 μmol; 73%). Note: After SPE, the tert-butyl protecting group was removed after radiofluorination, HPLC purification, and solvent evaporation by treatment with pure TFA at 95 °C for 3 min. LC-MS R t 16.02 min, m / z 1219.5858 [M+H] +

[0225] [ka] 2-(2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetoxy)acetic acid 28.99 mg (50.6 μmol) of tris-tBu-DOTA, 90.65 mg (278 μmol) of cesium carbonate, and 10.28 μL (15.0 mg; 65.5 μmol) of 2-bromobenzyl acetate were suspended in 300 μL of dimethylformamide and shaken for 2 hours. The product was isolated by HPLC, lyophilized, and dissolved in 25 mL of 10% acetic acid in methanol. 50 mg of 10% Pd / C and hydrogen (atmospheric pressure) were added. After 2 hours, the solvent was removed and the title compound was isolated by HPLC. After lyophilization, 25.19 mg (39.9 μmol; 79%) of the title compound was obtained. LC-MS R t 14.14 min, m / z 631.4784 [M+H] +

[0226] [ka] tBu-FAPI-79 2.00 mg (3.41 μmol) of (S)—N-(2-(2-cyano-4,4-difluoropyrrolidin-1-yl)-2-oxoethyl)-6-(3-(1-tert-butoxycarbonyl-piperidin-4-yl)-1-propoxy)quinoline-4-carboxamide was dissolved in 50 μL of acetonitrile and 100 μL of trifluoroacetic acid. After 10 min, the volatiles were removed and the residue was washed with diethyl ether. 4.20 mg (6.60 μmol) of 2-(2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl)acetoxy)acetic acid and 3.35 mg (8.84 μmol) of HBTU were dissolved in 100 μL of dimethylformamide, and 10.0 μL (7.40 mg; 57.4 μmol) of DIPEA was added to the dried residue and reacted for 60 min. After HPLC purification and lyophilization, 2.26 mg (2.06 μmol; 60%) of the title compound was obtained. LC-MS R t 12.98 min, m / z 1099.7481 [M+H] +

[0227] [ka] FAPI-79 2.26 mg (2.06 μmol) of tBu-FAPI-79 was dissolved in 25 μL of acetonitrile and 100 μL of trifluoroacetic acid and shaken at 35° C. for 30 minutes. After evaporation of the solvent, the product was isolated by HPLC. After lyophilization, 1.58 mg (1.70 μmol; 82%) of the title compound was obtained. LC-MS R t 8.84 min, m / z 466.2737 [M+2H] 2+

[0228] Compound analysis Reversed-phase high-performance liquid chromatography (RP-HPLC) was performed using a linear gradient of aqueous acetonitrile (0–100% acetonitrile in 0.1% TFA over 5 min at a flow rate of 2 mL / min) on a Chromolith Performance RP-18e column (100 × 3 mm; Merck KGaA, Darmstadt, Germany). UV-absorbance was detected at 214 nm. An additional γ-detector was used for HPLC analysis of radioactive compounds. HPLC-MS characterization was performed using an ESI mass spectrometer (Exactive, Thermo Fisher Scientific, Waltham, MA, USA) coupled to an Agilent 1200 HPLC system equipped with a Hypersil Gold C18 1.9 μm column (200 × 2.1 mm, 0–100% acetonitrile over 20 min at a flow rate of 200 μL / min). Analytical radio-HPLC was performed using a Chromolith Performance RP-18e column (100 × 3 mm; Merck; 0–30% acetonitrile per 10 min; flow rate 2 mL / min). HPLC purification was performed on a LaPrep P110 system (Knauer, Berlin, Germany) and a Reprosil Pur 120 column (C18-aq 5 μm 250 × 25 mm; Dr. Maisch, Ammerbuch-Entringen, Germany). The water / acetonitrile gradient (15 or 25 min; 0.1% TFA; flow rate 20 mL / min) was varied for each product.

[0229] Radiochemistry Radioactive iodine (I-125) was purchased from Hartmann Analytik (Goettingen, Germany); radioactive lutetium (Lu-177) was obtained from ITG (Munchen, Germany); radioactive gallium (Ga-68) was eluted from a Ge-68 / Ga-68 generator purchased from Themba Labs (Somerset West, South Africa). Tc-99m was eluted from a Mo-99 / Tc-99m generator (Curium Pharma, Berlin, Germany). Cu-64 was provided by UKT Tuebingen (Tuebingen, Germany). Sm-153 was provided by DSD Pharma (Purkersdorf, Austria). Pb-203 was provided by Lantheus (N. Billerica, MA, USA). F-18-FDG and F-18-fluoride were provided by ZAG Zyklotron AG (Eggenstein, Germany). The CRS kit for tricarbonyl was obtained from the Paul Scherrer Institut (Villingen-PSI, Switzerland).

[0230] For iodination, 10 μL of the organotin precursor of FAPI-01 (1 μmol / mL in ethanol) was diluted with 10 μL of 1 M hydrochloric acid and 10 μL of water, followed by the addition of 1-20 MBq of iodine-125 in 0.05 M NaOH. The reaction was initiated by adding 5 μL of a fresh 1.9% peracetic acid solution in glacial acetic acid. After 60 s, 15 μL of 1 M NaOH was added, and the reaction was stopped by adding 5 μL of 5% aqueous ascorbic acid, followed by HPLC purification. The resulting solution was used directly for in vitro experiments or, in the case of animal studies, evaporated to dryness under reduced pressure and taken up in 0.9% NaCl (Braun, Melsungen, Germany).

[0231] Cu-64, Lu-177, and Pb-203 labeling of DOTA-compounds was performed by adding 5 MBq of the radionuclide to 100 μL of a 10 μM solution of the respective precursor in 0.1 M NaOAc (pH 5) and incubating at 95 °C for 10 min. This solution was used directly for in vitro experiments or diluted with 0.9% NaCl (Braun, Melsungen, Germany) for biodistribution studies. For imaging studies in mice (scintigraphy, PET), the radiotracer was post-processed by solid-phase extraction (sep-pak light C18, Waters).

[0232] Prior to Tc(I) labeling, 1 mL of Tc-99m-pertechnetate in 0.9% saline was added to the CRS kit and incubated for 20 minutes. After cooling to room temperature, a mixture of 25.0 μL of precursor (1 mM in water), 150 μL of phosphate buffer (0.4 M, pH 7.4), and 240 μL of hydrochloric acid (1.0 M) was added, and the final mixture was adjusted to pH 5 as needed. The reaction was carried out at 95°C for 20 minutes and followed by solid-phase extraction (Sep-Pak Light C18, Waters). For in vivo and animal studies, labeling was performed after Tc(VII) reduction using one-fifth of the reagent and 200 μL of CRS kit solution.

[0233] Prior to Tc(V) labeling, 30 μL of SnCl2 solution containing 200 mM glucoheptonate was incubated with 200 μL of Tc-99m-pertechnetate in 0.9% saline for 10 min at room temperature. 5.00 μL of precursor (1 mM in water) and 3.75 μL of sodium hydroxide solution (0.1 M in water) were added, and the final mixture was incubated at 95 °C for 20 min. For mouse imaging studies (scintigraphy), the radiotracer was post-cleared by solid-phase extraction (sep-pak light C18, Waters).

[0234] Labeling with Ga-68 for animal studies was performed by incubating 255 μL of generator eluate (0.6 M HCl, approximately 230 MBq) with a mixture of 1 nmol of DOTA precursor, 1 μL of 20% aqueous ascorbic acid, and 72 μL of 2.5 M NaOAc at 95 °C for 10 min. Residual free radioactivity was removed by dilution with 2 mL of water, solid-phase extraction (Sep-Pak Light C18, Waters), washing with 2 mL of water, and elution with 1 mL of 1:1 water / ethanol. The resulting solution was evaporated to dryness under reduced pressure, and the residue was taken up in 0.9% NaCl (Braun).

[0235] For the formation of the AlF-NOTA complex, F-18 fluoride was trapped on a water Sep-Pak QMA plus light cartridge (46 mg sorbent, pretreated with 0.5 M NaOAc, pH 3.9), washed with water, and eluted with 500 μL of 0.1 M NaOAc (pH 3.9). For animal studies, 150 μL of the eluate was preincubated with 2 μL of AlCl3 solution (10 mM in water) and 50 μL of DMSO. After 5 min, this mixture was added to 40 nmol of NOTA precursor (10 μL of a 4 mM solution in water) and 1 μL of 20% ascorbic acid in water. The solution was reacted at 95 °C for 15 min. The product was isolated by HPLC (0-20% acetonitrile per 10 min), the solvent removed, and taken up in 0.9% saline prior to injection.

[0236] For the formation of 6-fluoronicotinamide, F-18 fluoride was trapped on a Waters Sep-Pak QMA plus light cartridge (46 mg of sorbent; pretreated with 0.5 M KHCO), washed with water, dried, and eluted with a mixture of 7.50 mg (19.9 μmol) of cryptofix 222 and 1.99 mg (1.99 μmol) of KHCO in 450 μL of acetonitrile and 50 μL of water. After solvent removal, the residue was dried by azeotropic distillation with 3 × 1 mL of acetonitrile. This residue was taken up in 100 μL of 1:1 tert-butanol / acetonitrile and added to 1 mg (approximately 1.3 μmol) of trimethylpyridin-2-aminium precursor. The solution was reacted at 75 °C for 10 min. The product was isolated by HPLC (0–30% acetonitrile per 10 min), solvent removed, and taken up in 0.9% saline prior to injection.

[0237] Alternatively, 6-fluoronicotinamide was synthesized by trapping F-18 fluoride on a Waters Sep-Pak QMA Plus Light cartridge (46 mg of sorbent; pretreated with 0.5 M KHCO3), washing with acetonitrile, drying, and eluting with 0.5 mg (approximately 0.4–0.6 μmol) of the (protected) FAPI precursor in 0.5 mL of tert-butanol. The solvent was removed under reduced pressure, and the residue was taken up in 100 μL of 1:4 acetonitrile / tert-butanol. After 20 min at 70 °C, the reaction mixture was diluted with water, and the protected intermediate was worked up by solid-phase extraction (Sep-Pak Light C18, Waters). The solvent was removed, and 200 μL of trifluoroacetic acid was added to the residue. The mixture was heated to 95 °C for 3 min, dried under reduced pressure, and diluted with water. After this, the product was isolated by HPLC, which was performed directly on the diluted reaction mixture in the case of compounds lacking a protecting group. For animal studies, the product was removed from the solvent and taken up in 0.9% saline prior to injection (uncorrected radiochemical yield approximately 25%).

[0238] To measure stability in human serum, radiolabeled compounds (approximately 2.5 MBq for I-125 and approximately 15 MBq for Lu-177) were purified (HPLC or solid-phase extraction) and the solvent removed. The residue was taken up in 250 μL of human serum (Sigma-Aldrich) and incubated at 37 °C. Samples were precipitated with 30 μL of acetonitrile and analyzed by HPLC (0-30% acetonitrile per 10 min).

[0239] Example 2: In vitro characterization of FAPI derivatives In vitro binding studies were performed using the human tumor cell lines BxPC3, Capan-2, MCF-7 (purchased from Sigma-Aldrich Chemie GmbH), and SK-LMS-1 (purchased from ATCC) as well as the stably transfected FAP cell lines HT-1080-FAP, HEK-muFAP, and the CD26-expressing cell line HEK-CD26 (obtained from Stefan Bauer, NCT Heidelberg). All cells were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal bovine serum at 37°C under 5% carbon dioxide. For fluorescence internalization experiments, cells were seeded on glass coverslips and stained with FAPI-02-Atto488 and DAPI for nuclear staining. Images were acquired using a laser scanning confocal microscope with a 63x oil immersion objective. Radioligand binding studies were performed using HT-1080-FAP cells. Radiolabeled compounds were added to the cell cultures and incubated for different time intervals ranging from 10 min to 24 h. Competition experiments were performed using unlabeled compounds (10 -5 M~10 -9 Experiments were performed by simultaneously exposing cells to 1 M glycine-HCl (1 M glycine-HCl) and radiolabeled compound for 60 min. In efflux experiments, the radioactive medium was removed after 60 min of incubation and replaced with non-radioactive medium for time intervals ranging from 1 to 24 h. In internalization experiments, surface-bound activity was removed by incubating cells with 1 M glycine-HCl buffer for 10 min. Radioactivity was measured using a γ-counter, normalized to 1 mio cells, and calculated as a percentage of the applied dose (%ID).

[0240] Cell staining and microscopy For internalization experiments, HT-1080-FAP cells and HEK muFAP cells were seeded on uncoated coverslips in 24-well plates and cultured in 10% fetal bovine serum-containing medium until they reached approximately 80-90% confluence. The medium was removed and the cells were washed twice with 0.5 mL of PBS, pH 7.4. FAPI-02-Atto488 (20 μM in DMEM) was added to the cells and incubated at 37°C for 2 hours. The cells were washed three times with 0.5 mL of PBS, pH 7.4, and fixed with paraformaldehyde (2% in PBS) for 15 minutes. The overgrown coverslips were placed on glass slides using DAPI-containing mounting medium for nuclear staining (Fluoroshield, Sigma-Aldrich). Images were acquired on a laser scanning confocal microscope (Zeiss LSM 700; Zeiss, Oberkochen, Germany) with a 0.099 x 0.099 μm x y pixel setting and a pinhole size of 1 Airy unit using a Zeiss Plan-Apochromat 63x / 1.4 Oil DIC III immersion objective for each fluorophore used (488 nm for FAPI-02-Atto488 and 405 nm for DAPI). Photographs were processed consistently using Z ZEN 2008 software and ImageJ.

[0241] Radioligand binding assay For radioligand binding studies, cells were seeded into 6-well plates and cultured for 48 hours to reach a final confluence of approximately 80-90% (1.2-2 mio cells / well). The medium was replaced with 1 mL of fresh medium without fetal bovine serum. Radiolabeled compounds were added to the cell culture and incubated for different time intervals ranging from 10 minutes to 24 hours. Competition experiments were performed using unlabeled (10 -5 M~10 -9The experiments were performed by simultaneously exposing cells to 100 μg / ml of PBS and radiolabeled compound for 60 min. In efflux experiments, the radioactive medium was removed after 60 min of incubation and replaced with non-radioactive medium for time intervals ranging from 1 to 24 h. In all experiments, cells were washed twice with 1 mL of phosphate-buffered saline, pH 7.4, followed by lysis with 1.4 mL of lysis buffer (0.3 M NaOH, 0.2% SDS). Radioactivity was measured using a counter (Cobra II, Packard), normalized to 1 mio cells, and calculated as a percentage of the applied dose (%ID). Each experiment was performed in triplicate, with each independent experiment repeated three times.

[0242] For internalization experiments, cells were incubated with radiolabeled compounds for 60 min at 37°C and 4°C. Cellular uptake was terminated by removing the medium from the cells and washing them twice with 1 mL of PBS. Subsequently, cells were incubated with 1 mL of glycine-HCl (1 M in PBS, pH 2.2) at room temperature for 10 min to remove surface-bound activity. Cells were washed with 2 mL of ice-cold PBS and lysed with 1.4 mL of lysis buffer, and the internalized fraction was measured. For cells incubated at 4°C, all washing and elution steps were performed with ice-cold buffer. Radioactivity was measured using a γ-counter, normalized to 1 mio cells, and calculated as a percentage of the applied dose (%ID).

[0243] FAPI-01 selectively targets human and mouse FAP-α. To analyze the binding properties of FAPI-01 to its target proteins, radioligand binding assays were performed using different cancer cell lines and cell lines transfected with human and mouse FAP and CD26, a closely related membrane protein also known as DPPIV. Both mouse FAP and CD26 show high homology with human FAP-α (muFAP: 90% identity and 94% similarity at the amino acid level; CD26: 52% identity and 71% high structural similarity) (Kelly T., Drug Resist Update, 2005).

[0244] As shown in Figure 1A, FAPI-01 did not significantly bind to FAP-negative cancer cell lines and targeted human and mouse FAP-α-expressing cells with high affinity (IC 50 Human FAP-α = 39.4 nM). Furthermore, no substantial binding to CD26-expressing cells was observed (0.05 ± 0.01%), demonstrating that FAPI-01 selectively targets FAP-α. This is particularly important because CD26 is highly expressed in various normal tissues, including the kidney, liver, and small intestine. The high selectivity of the ligand for FAP-α is a major advantage in avoiding high background signals due to nonspecific CD26 binding, resulting in optimal image quality.

[0245] FAPI-01 was rapidly internalized in FAP-positive cells but exhibited time-dependent efflux and robust deiodination. Cell-based internalization assays demonstrate rapid cellular uptake of FAPI-01 (Figure 1B). After 10 minutes of incubation, 95% of the total bound fraction was located intracellularly (19.70 ± 0.28% total). Over a 4-hour period, only limited decline in activity was observed (17.00 ± 0.40% total, of which 94% was internalized).

[0246] Iodine-labeled compounds often exhibit time-dependent enzymatic deiodination. This was also observed with FAPI-01, resulting in a decrease in the intracellular radioactivity of this compound after longer incubation times (3.25 ± 0.29% after 24 h). Deiodination could be minimized by lowering the temperature to 4°C, which reduced the activity of the deiodinase enzyme, resulting in a 26.66 ± 1.59% increase in radioactivity after 24 h.

[0247] FAPI-02 shows enhanced binding and uptake to human FAP-α compared to FAPI-01. To avoid the rapid loss of activity of FAPI-01 due to enzymatic deiodination, we designed a non-halogenated derivative, FAPI-02, in which the FAP-binding moiety was chemically linked to the chelator DOTA. This modification resulted in enhanced stability and offered the possibility of easily incorporating either diagnostic or therapeutic radionuclides, enabling the use of FAPI-02 as a non-flammable compound. Similar to its iodinated analog, FAPI-02 inhibited the activity of human and mouse FAP-(IC) receptors without addressing CD26. 50 FAPI-02 specifically binds to human FAP-α (21 nM)-expressing cells (%ID = 0.13 ± 0.01%; Figure 1A). FAPI-02 is rapidly internalized into FAP-α-expressing cells (20.15 ± 1.74% ID after 60 min, of which 96% was internalized; Figure 1B), indicating a more stable and higher uptake rate over time. Compared to FAPI-01 binding after 10 min of incubation, only 5% of the activity remained after 24 h. Meanwhile, 34% of the initial radioactivity of FAPI-02 was detected after 24 h of incubation. Efflux experiments demonstrated that FAPI-02 is cleared significantly slower than FAPI-01, retaining 12% of the initially accumulated radioactivity after 24 h (FAPI-01 retains 1.1% ID after 24 h; Figure 1E).

[0248] We confirmed robust internalization of FAPI-02 into human and mouse FAP-α-expressing cells by fluorescence laser scanning microscopy. To this end, HT-1080-FAP and HEK-muFAP cells were stained with a fluorescently labeled FAPI-02 derivative (FAPI-02-Atto488) for 1–2 h. As shown in Figure 1D, the compound was completely internalized and accumulated inside FAP-α-expressing cells, whereas no uptake was detected in FAP-α-negative HEK-CD26 cells.

[0249] Design of FAPI derivatives with improved binding properties and pharmacokinetics Further variants of FAPI-02 were designed to increase tumor retention time, aiming to develop theranostic FAP-targeting agents. Variants FAPI-03 to FAPI-15 were characterized for target binding, internalization rate, and target specificity. The results are shown in Figure 2.

[0250] Example 3: PET imaging and biodistribution analysis in mice All experiments were performed in accordance with the German Animal Protection Act and conformed to the European Commission regulations for the care and use of laboratory animals. Mice were anesthetized by isoflurane inhalation.

[0251] For in vivo experiments, 8-week-old BALB / c nu / nu mice (Charles River) were subcutaneously inoculated with HT-1080-FAP, Capan-2, or SK-LMS-1 cells at 5 × 106 in the right trunk. Tumors measuring approximately 1 cm were obtained. 3 When the PET imaging dose reached 10 MBq, the radiolabeled compound was injected via the tail vein (approximately 10 MBq for small animal PET imaging and approximately 1 MBq for organ distribution). PET imaging was performed up to 140 min after intravenous injection of approximately 1 MBq of Ga-68 labeled compound per mouse using an Inveon PET small animal PET scanner (Siemens). Images were iteratively reconstructed using 3D-OSEM+MAP and converted to standardized uptake value (SUV) images. Quantification was performed using ROI techniques and expressed as SUVmean. For organ distribution of Lu-177 labeled compound (approximately 10 MBq per mouse), animals (n = 3 per time point) were sacrificed after the indicated time points (30 min to 24 h). Distributed radioactivity was measured in all dissected organs and blood using a γ-counter (Cobra Autogamma, Packard). Values ​​were expressed as the percentage of the injected dose per gram of tissue (%ID / g).

[0252] In the pharmacokinetic modeling, the transport constant K1 and the rate constants k2-k4 were calculated using a two-tissue compartment model implemented in PMOD software [4], taking into account the vascular fraction (vB) related to the volume of blood exchanging with tissue within the VOI. The rate constants describing the compartmental flux include k1 (binding to receptors), k2 (shedding), and k3 (internalization) and k4 (efflux) within the tumor tissue. In this model, the fractional volume of distribution (DV = K1 / k2) is given by 15 is the percentage of the region of interest that is filled with O-labeled water.

[0253] FAP variants also accumulate in human FAP-expressing and non-FAP-expressing xenografts due to recruitment and activation of mouse fibroblasts. Tumor accumulation of FAPI-02 and -04 was assessed by small-animal PET imaging in mice bearing xenografts from both human FAP-α-positive and -negative tumor cells. In both cases, the radiotracer rapidly concentrated within the tumor and remained concentrated for at least 140 min (Figure 3A, C, E, G). At the same time, FAPI-02 and -04 demonstrated negligible nonspecific binding and were rapidly cleared from the blood, primarily via the kidneys and bladder, resulting in low background and beneficial tumor-to-organ ratios. Coadministration of unlabeled compounds as competitors resulted in a complete absence of radioactivity in the tumor, demonstrating the specificity of the radiotracer for its target protein (Figure 4). Interestingly, high tumor uptake of FAPI-02 was observed in mice bearing FAP-α-positive (HT-1080-FAP) and FAP-α-negative (Capan-2) tumor cell lines, due to the recruitment and activation of activated mouse fibroblasts. The pharmacokinetic properties of the radiotracers calculated from the PET data using a two-tissue compartment model according to Burger et al., Nucl Med, 1997, are shown in Table 6. [Table 6] Table 6. Calculated from PET data using a two-tissue compartment model according to Burger et al., Nucl Med, 1997 68Pharmacokinetic properties of Ga-FAPI-02. vB: vascular fraction related to the volume of blood exchanging with tissues within the VOI (volume of interest); k1-k4: calculated rate constants; Vs: ratio of specific binding concentration to total parent at equilibrium; Vt: total volume of distribution

[0254] These observations suggest that biodistribution studies 177 This was confirmed with Lu-FAPI-02 and -04, which demonstrated rapid tumor accumulation in both human FAP-α positive and negative tumors (see Table 7 for quantified uptake values), along with very low activity in all other organs, resulting in beneficial tumor-to-organ ratios (Figure 5D-F) in HT-1080-FAP tumor-bearing mice. 177 Similar results were obtained with Lu-FAPI-04. Compared to FAPI-02, FAPI-04 exhibits higher tumor uptake, especially at 24 hours (Figure 5C). Area under the curve (AUC) calculations are shown in Table 8. [Table 7] Table 7. Quantification of biodistribution data 1 hour after intravenous administration of Lu-177 labeled FAPI-02 and -04 to tumor-bearing Balb / c nude mice; n=3; values ​​reported as mean %ID / g ± SD [Table 8] Table 8. Tumor uptake of selected FAPI derivatives in nude mice bearing HT-1080-FAP tumors, n=3. Values ​​are reported as mean (ID / g±SD).

[0255] Example 4: Clinical PET / CT Study Imaging studies of more than 100 patients were performed for medical reasons in accordance with § 13(2b) of the German Medicines Act under the conditions of the update of § 37 of the Declaration of Helsinki (unproven interventions in clinical practice). 68 Experiments were performed using either Ga-FAPI-02 or -04, and the tracer was administered intravenously (20 nmol, 122-336 MBq) at 10 min, 1 h, and 3 h after administration. The activity of the injected radioactive tracer varied. 68The short half-life of Ga 68 Ge / 68 This is due to the variable elution efficiency obtained during the life of the Ga generator. 18 FDG imaging was performed 1 hour after intravenous injection of 358 MBq of F-FDG. PET / CT scans were performed using a Biograph mCT Flow TM The PET / CT scan was performed on a Siemens Medical Solution PET / CT scanner using the following parameters: slice thickness 5 mm, increment 3–4 mm, soft tissue reconstruction kernel, and care dose. Immediately after the CT scan, whole-body PET was performed using FlowMotion. TM The data were acquired at 0.7 cm / min in 3D (matrix 200x200). Emission data were corrected for random scattering, scattering, and attenuation. Reconstruction was performed with an ordered subset expectation maximum (OSEM) algorithm with 2 iterations / 21 subsets and Gaussian filtered to a transverse resolution of 5 mm at full width at half maximum (FWHM). Attenuation correction was performed using low-dose unweighted CT data. Quantitative evaluation of standardized uptake values ​​(SUVs) was performed using region-of-interest techniques.

[0256] FAPI-02 and -04 rapidly accumulate in human metastatic breast, pancreatic, lung, HNO, small intestine, and ovarian cancers. For patients with metastatic breast cancer, lung cancer, pancreatic cancer, HNO cancer, small intestine cancer, and ovarian cancer 68 Diagnostic PET / CT scans were performed 1 hour after intravenous administration of Ga-FAPI-02 and -04. In all patients, robust tracer accumulation was observed in the primary tumor as well as in lymph nodes and bone, with a maximum SUV value of 48.0 in metastases. In contrast, tracer uptake in normal tissues was very low (Figures 6-14). Radioactivity was rapidly cleared from the bloodstream and mostly excreted via the kidneys, resulting in high-contrast images. Comparative imaging in one patient with locally advanced lung adenocarcinoma demonstrated the efficacy of commonly used PET tracers. 18Compared with F-FDG, a clear advantage of FAPI-02 was revealed. As shown in Figure 9, FAPI-02 exhibited high uptake with lower background activity, which translates into higher contrast for better visibility of metastatic lesions. Unlike FDG, which accumulates in high glucose-consuming cells such as the brain, FAPI-02 selectively targets tissues where FAP-α is expressed. Comparative imaging in one prostate cancer patient demonstrated a significant improvement over commonly used PET tracers. 68 Ga-DOTATOC and 68 Compared to Ga-PSMA, a clear advantage of FAPI-04 emerged, which allowed for the detection of smaller tumor lesions with reduced tracer accumulation in the kidney (Figure 14).

[0257] Consideration Reliable diagnosis of primary tumors, metastatic lesions, and involved lymph nodes is paramount to enable effective and appropriate therapy planning, including tumor staging and treatment selection. To this end, imaging techniques are essential tools for the diagnosis of many types of cancer. PET / CT combination is the method of choice for modern tumor diagnosis due to its high diagnostic accuracy and the possibility of assessing both anatomical and physiological details. However, in contrast to noninvasive imaging techniques such as magnetic resonance imaging (MRI) or CT alone, PET / CT combination requires the use of radiotracers with high affinity for target structures whose expression is enhanced in tumors compared with normal tissues. An ideal tracer should bind specifically to its target protein, ensuring reliable differentiation between cancerous and healthy tissues and low background signal, resulting in high-contrast images. Affinity and specificity become even more important when the radiotracer represents a theranostic compound, i.e., offering the possibility of carrying either diagnostic or therapeutic nuclides, which facilitate and improve targeted and personalized treatment. For potential applications of tracers for therapeutic purposes, high target specificity ensures reduced side effects, which is particularly important for the protection of radiosensitive tissues such as bone marrow, reproductive and digestive organs.

[0258] With this in mind, we developed a theranostic tracker that targets cancer-associated fibroblasts, which form a major component of the tumor stroma. They are known to play a key role in tumor growth, metastasis, and progression, and are genetically more stable than cancer cells, making them less susceptible to the development of therapy resistance. In contrast to normal fibroblasts, CAFs express specific proteins that can be used as tumor-specific markers. These include the membrane protein FAP-α, which is widely expressed in various tumor microenvironments, enabling targeting of various tumor entities, including pancreatic, breast, and lung cancers, which account for a large proportion of all solid tumors.

[0259] Based on this high-affinity small molecule enzyme inhibitor for the target protein, we developed radiotracers FAPI-01 through FAPI-73 through focused chemical modification. All compounds demonstrate specific binding to human and mouse FAP-α with rapid and nearly complete internalization, without addressing the closely related protein CD26 / DPP4. Because the iodinated molecules undergo enzymatic deiodination with the release of free iodine, prolonged incubation times result in low intracellular radioactivity. For this reason, FAPI-02 and subsequent compounds were designed so that the FAP-binding moiety is chemically conjugated to the chelator DOTA. This resulted in a series of theranostic compounds with favorable pharmacokinetic and biochemical properties, of which FAPI-02, FAPI-04, FAPI-46, FAPI-34, FAPI-42, FAPI-52, FAPI-69, FAPI-70, FAPI-71, FAPI-72, and FAPI-73 are the most preferred ligands. Both FAPI-02 and FAPI-04 were eliminated significantly slower than FAPI-01, retaining 12% (FAPI-02) and 49% (FAPI-04) of the original accumulated radioactivity after 24 hours (FAPI-01: 1.1%), while other preferred compounds possess even stronger binding avidity (Figure 16). They are rapidly internalized into FAP-α-expressing cells, demonstrating high tumor uptake rates in both tumor-bearing mice and patients with metastatic epithelial cancer. In contrast, there is no accumulation in normal tissues, and rapid clearance from the blood system allows for high-contrast imaging. Robust internalization into both human and mouse FAP-α-expressing cells was confirmed by confocal microscopy using fluorescently labeled FAPI-02. In contrast to the first-generation FAP antibody F19, which has high affinity for its target protein without being internalized, FAPI-02 demonstrates complete cellular uptake after 1 hour of incubation. The mechanism of internalization after FAP binding was studied by Fischer et al. using FAP antibody fragments (Fabs) and DyLight 549 anti-mouse antibody in SK-Mel-187 cells. Incubation at 37°C resulted in internalization of the FAP-antibody complex.Similar to our small molecules, the internalization process occurred rapidly, with nearly complete internalization. Colocalization of the Fab with markers for early endosomes was observed after 20 minutes, and with markers for late endosomes and lysosomes after 40 minutes. Fab-mediated FAP-α internalization was suppressed by inhibitors of dynamin-dependent endocytosis, indicating that endocytosis occurs via a dynamin-dependent mechanism.

[0260] FAPI-02 and -04 are not retained in the renal parenchyma and are rapidly removed from the organism by renal clearance. They accumulate in inflamed tissues or in cells with high glucose consumption, including the brain. 18 Unlike F-FDG, FAPI-02 is selectively enriched in tissues where the target protein is expressed. This opens up new perspectives for detecting malignant lesions in these areas. Furthermore, FAP-α has also been shown to be expressed by rheumatoid myofibroblast-like synoviocytes in patients with rheumatoid arthritis and osteoarthritis, atherosclerosis, and fibrosis, as well as in ischemic cardiac tissue after myocardial infarction. These observations suggest the application of FAPI-02 and -04 as imaging tracers for further indications.

[0261] The limiting factor for tumor lesion detection is the degree of FAP-α expression within the tumor. This is largely determined by the number of activated fibroblasts, i.e., the stromal content, and / or the number of FAP-α molecules per fibroblast, which can be determined by the microenvironment. Because tumor growth exceeding 1–2 mm in size fundamentally requires the formation of supporting stroma, it is possible to visualize small lesions in the 3–5 mm range using FAPI-PET / CT.

[0262] As with other targeted approaches, FAPI derivatives achieve optimal results only in tissues with sufficiently high FAP-α expression, which is known to be highly heterogeneous among different cancer types and patients. Besides breast, colon, and pancreatic cancers, which are excellent candidates for FAPI imaging, it remains to be explored whether other tumor entities, such as lung, head and neck, ovarian, and hepatoma, are also favorable targets.

[0263] Furthermore, FAP-α expression was demonstrated in wound healing and fibrotic tissue, which should be kept in mind when interpreting radiological findings. These facts highlight the need to properly assess which patients are likely to benefit from potential FAPI therapy. Given the ability to use either diagnostic or therapeutic nuclides, FAPI-02 and -04 allow for easy stratification of appropriate patient cohorts. In any event, it is already clear that both FAPI tracers are ideal candidates for the development of targeted radiopharmaceuticals. Their high targeting affinity, rapid tumor internalization, and rapid systemic clearance already make them ideally suited for tumor imaging.

[0264] Example 5: Characterization of FAPI in vitro and in vivo Experimental methods and clinical evaluation All in vitro and in vivo experiments, as well as the clinical evaluation of FAPI derivatives, were performed as described above by Loktev et al. 1 and Lindner et al. 2 Preliminary dose estimates for FAPI-02 and FAPI-04 were based on two patients examined at 0.2, 1, and 3 hours after tracer injection using the QDOSE dose estimation software suite. Additional PET / CT scans for tumor patients were acquired 1 hour after injection of either FAPI-02 (n = 25) or FAPI-04 (n = 25); for six patients, intrapersonal FDG scans (also acquired 1 hour later) were available. For normal tissues of 16 organs, a 2-cm spherical VOI was placed in the parenchyma, and for tumor lesions, a thresholded segmented VOI was used to measure SUVmean / max. 3 was quantified.

[0265] In vitro characterization of DOTA-FAPI derivatives To evaluate the target binding and internalization rates of DOTA-FAPI derivatives compared to FAPI-04, Lu-177-labeled compounds were incubated with FAP-expressing HT-1080 cells for 1, 4, and 24 hours, respectively (Figure 16). The membrane-bound fraction was removed by acidic elution with glycine-HCl at pH 2.2, followed by alkaline cell lysis and measurement of the internalized fraction. As shown in Figure 16, all derivatives exhibited higher cell binding compared to FAPI-04, with binding values ​​reaching 500% of the maximum lead compound after 1 hour of incubation (up to 750% after 4 hours).

[0266] To assess target affinity and specificity, competitive binding assays were performed using increasing concentrations of unlabeled compounds as competitors for the Lu-177 labeled compounds (Figure 17; respective IC values ​​listed in Table 9). 50 value). [Table 9] Table 9. IC of selected FAPI derivatives measured by competitive binding assay 50 value

[0267] Organ distribution of DOTA-FAPI derivatives in tumor-bearing mice To analyze the pharmacokinetic profile and in vivo tumor uptake, Lu-labeled DOTA-FAPI derivatives were intravenously administered to HT-1080-FAP tumor-bearing mice. The organ distribution of the radiolabeled compounds was measured ex vivo in blood, healthy tissue, and tumor. As shown in Figure 19, most compounds show higher tumor uptake rates compared to FAPI-02 and FAPI-04, especially at 24 hours post-administration. Due to increased lipophilicity, some radiotracers show higher blood activity and increased kidney retention. Measurement of tumor-to-blood ratios reveals a clear advantage for compounds FAPI-21 and FAPI-46, which consistently show significantly higher ratios than FAPI-04 in our experiments (Figure 20).

[0268] Small animal imaging of DOTA-FAPI derivatives in tumor-bearing mice Based on these findings, we performed small-animal PET imaging up to 140 minutes after intravenous administration of a Ga-68-labeled DOTA-FAPI derivative as a radiotracer to HT-1080-FAP tumor-bearing mice. The favorable tumor-to-blood ratios of FAPI-21 and FAPI-46 resulted in high-contrast images, enabling excellent visualization of FAP-positive tumors (Figure 21). Quantitative analysis of tracer accumulation in tumor, kidney, liver, and muscle tissues (given as SUVmax values) indicates slightly lower muscle, kidney, and liver activity for FAPI-46 compared to FAPI-21 (Figure 22).

[0269] Biodistribution and dosimetry estimates of FAPI-02 and FAPI-04 compared with FDG in cancer patients Similar to literature values ​​for F-18-FDG, Ga-68-DOTATATE, or Ga-68-PSMA-11, 200 MBq of Ga-68-FAPI-02 and -04 corresponds to an equivalent dose of approximately 3-4 mSv in our experiments. After rapid clearance via the kidneys between 10 minutes and 3 hours after injection, normal organs show low tracer uptake with minimal changes. For FAPI-02, tumor uptake is reduced by 75% between 1 and 3 hours after injection, whereas tumor retention is slightly prolonged for FAPI-04 (50% washout). At 1 hour after injection, tumor uptake of both FAPI-tracers is equivalent (Figure 23). Compared to FDG, tumor uptake was nearly equal (mean SUVmaxFDG 7.41; SUVmaxFAPI-02 7.37; ns); background uptake was significantly lower with FAPI-02 in the brain (11.01 vs 0.32), liver (2.77 vs 1.69), and oral / pharyngeal mucosa (4.88 vs 2.57); there were no relevant differences between FDG and FAPI-02 in other organs (Figure 24). For detailed information and results, see Giesel et al., which is incorporated herein by reference. 3 See

[0270] PET imaging of FAPI-04 in patients with various cancers and non-cancerous malignancies In addition to the rapid uptake of Ga-68-labeled FAPI-04 in various cancers, including breast, pancreatic, ovarian, and HNO tumors, tracer accumulation was also demonstrated in several inflammatory malignancies, such as peritonitis carcinomatosa (Figure 25A) and myocarditis (Figure 25B) and arthropathy (Figure 25C). These results demonstrate the potential application of Ga-68-labeled FAPI for the detection of noncancerous malignancies characterized by chronic inflammatory processes involving the recruitment of activated fibroblasts.

[0271] PET imaging of FAPI-21 and FAPI-46 in patients with various cancers As shown in Figure 26, robust accumulation of Ga-68-labeled FAPI-21 was observed in a variety of cancers, including ovarian, rectal, and mucoepidermoid carcinomas. Similar tumor uptake was observed with Ga-68-labeled FAPI-46, which rapidly accumulated in cholangiocellular and colorectal carcinomas, lung cancers, and solitary fibrous sarcomas (Figure 27). Following PET / CT studies with Ga-68-labeled FAPI-46, the first therapeutic approach using a Sm-153-labeled radiotracer was performed in two cancer patients. As shown in Figure 28, robust tumor accumulation of the tracer was detectable up to 20 hours after administration. FAPI-46-PET / CT imaging of three lung cancer patients with idiopathic pulmonary fibrosis demonstrated clear differences in tracer accumulation between cancerous and fibrotic lesions. As shown in Figure 30, tumor uptake of Ga-68-labeled FAPI-46 was significantly higher in two patients (A, B) but slightly lower in one patient (C) compared with activity measured in fibrotic tissue. The patient shown in Figure C had exacerbated pulmonary fibrosis compared with two non-exacerbated cases. Therefore, this tracer may be useful for distinguishing between fibrotic patients with poor prognosis and those with good prognosis.

[0272] with alternative radionuclides, e.g., Tc-99m, Pb-203, Cu-64, and F-18 radioactive label FAPI derivatives for To enable the use of alternative radionuclides, a series of FAPI derivatives have been designed and characterized with respect to target affinity, specificity, and pharmacokinetics. In some of these compounds, the original chelator DOTA has been replaced by a different chelating moiety ideally suited for the uptake of Tc-99m (FAPI-19, -27, -28, -29, -33, -34, -43, -44, -45, -60, -61, -62). In vitro FAP affinity and biodistribution in HT-1080-FAP xenografted mice have been demonstrated for FAPI-19 and FAPI-34. Both compounds demonstrate robust binding to human FAP in vitro (IC). 50 FAPI-19: 6.4 nM). In contrast to FAPI-19, which showed poor tumor uptake in vivo and rapid liver accumulation due to renal excretion, FAPI-34 was continuously concentrated in tumors and showed significantly less liver uptake (Figures 31 and 32). The first diagnostic application of Tc-99m-labeled FAPI-34 in a patient with pancreatic cancer and liver metastasis demonstrated stable tumor accumulation of the tracer up to 4 hours after administration. Furthermore, the relatively low overall background radioactivity allowed for high-contrast imaging (Figure 33). This allows for broad application for scintigraphic and therapeutic diagnosis after labeling with Re-188.

[0273] Pb-203 radiolabeled FAPI derivatives (FAPI-04, -32, -46, and FAPI-04tcmc) showed comparable cellular binding to HT-1080-FAP cells, with FAPI-32 and FAPI-04tcmc reaching the highest binding values ​​after 60 min of incubation (26.93 ± 0.846 and 21.62 ± 0.61%ID / 1 mio cells, respectively, Figure 34A). While FAPI-32 was rapidly eliminated from tumor cells upon initial binding (t = 2 h), FAPI-04tcmc not only exhibited a significantly slower cellular efflux (t = 7 h) but also exhibited a significantly slower cellular efflux in competition assays (IC 50= 5.7 μM, Figure 34C). For this reason, we selected the optimal half-life and IC for further analysis in vivo. 50 FAPI-04 and FAPI-46 were selected for characterization. As shown in Figure 35, both compounds demonstrate negligible binding to healthy tissue but are continuously enriched within tumors. The scintigraphic findings are confirmed by biodistribution studies, where both radiotracers demonstrate robust tumor uptake, low overall organ activity, and rapid renal excretion (Figure 36).

[0274] To enable radiolabeling with Cu-64, NOTA derivatives FAPI-42 and FAPI-52 have been developed and characterized with respect to target affinity, specificity, and pharmacokinetics. As shown in Figure 37, both tracers have similar IC in the lower nanomolar range. 50 These compounds demonstrate robust binding to HT-1080-FAP cells for up to 24 hours of incubation at 1000 kJ / mL (Figure 37A, B). Nevertheless, FAPI-42 is eliminated significantly more slowly than FAPI-52, with a calculated in vitro half-life of 12 hours (Figure 37C). These results are confirmed by small-animal imaging of HT-1080-FAP xenografted mice. As shown in Figure 38, both compounds demonstrate robust tumor uptake and rapid clearance from the in vivo bloodstream. Notably, renal excretion of FAPI-42 is significantly faster than that of FAPI-52, yet its antitumor activity remains slightly higher between 2 and 24 hours after administration.

[0275] The NOTA derivatives FAPI-42 and FAPI-52 were configured to form aluminum fluoride complexes that enable imaging with F-18. As shown in Figure 39, both compounds demonstrate rapid tumor uptake in small-animal imaging of HT-1080-FAP xenografted mice. Both compounds are primarily excreted via the renal route, although biliary excretion has also been observed. While renal excretion is faster for FAPI-52, greater tumor accumulation, longer tumor residence time, and a lower proportion of biliary excretion favor FAPI-42.

[0276] References 1 Loktev, A. et al. A new method for tumor imaging by targeting cancer associated fibroblasts. Journal of nuclear medicine : official publication, Society of Nuclear Medicine, doi:10.2967 / jnumed.118.210435 (2018). 2 Lindner, T. et al. Development of quinoline based theranostic ligands for the targeting of fibroblast activation protein. Journal of nuclear medicine : official publication, Society of Nuclear Medicine, doi:10.2967 / jnumed.118.210443 (2018). 3 Giesel, F. et al. FAPI-PET / CT: biodistribution and preliminary dosimetry estimate of two DOTA-containing FAP-targeting agents in patients with various cancers. Journal of nuclear medicine : official publication, Society of Nuclear Medicine, doi:10.2967 / jnumed.118.215913 (2018).

[0277] Example 6: In vitro and in vivo FAPI characterization Preclinical Data Initial experiments were conducted in tumor-bearing mice using the human glioblastoma xenograft model U87MG with the aim of selectively targeting FAP-positive brain tumors. The tumor accumulation and organ distribution of radiolabeled FAPI-02 and -04 were analyzed by small-animal PET imaging and biodistribution studies. As shown in Figures 40 and 41, both FAPI-02 and -04 demonstrated rapid tumor uptake and negligible activity in healthy organs and blood.

[0278] Clinical Data According to the 2016 WHO classification, gliomas are subdivided into IDH wild-type gliomas (WHO grades I to IV) and IDH mutant gliomas (WHO grades II to IV). The most common WHO grade IV glioma is glioblastoma. Clinical PET imaging was performed on 18 patients with glioma (5 with IDH-mutant gliomas and 13 with IDH-wild-type glioblastomas; see Table 10). As shown in Figures 42-44, tracer uptake was demonstrated in IDH-wild-type glioblastomas and grade III / IV IDH-mutant gliomas, but not in grade II. In glioblastomas, spots of increased uptake were observed in projections onto the contrast-enhanced regions.

[0279] conclusion Increased tracer uptake in IDH wild-type glioblastoma and high-grade IDH-mutant astrocytomas, but not in diffuse astrocytomas, may enable noninvasive differentiation of low-grade IDH-mutant from high-grade gliomas and may be useful for follow-up studies. Heterogeneous tracer uptake in glioblastoma may aid in biopsy planning. [Table 10]

[0280] Example 7: In vitro and in vivo characterization of FAPI Reuptake experiments In the reuptake experiment, 177 Lu-labeled FAPI-04 and -46 (5 MBq / nmol in DMEM) were added to HT-1080-FAP cells and incubated for 60 min at 4 °C and 37 °C, respectively. The radioactive medium was removed, and the cells were washed twice with phosphate-buffered saline (PBS) at pH 7.4. Subsequently, non-radioactive medium with or without unlabeled FAPI (1 μM) was added for time intervals ranging from 10 min to 6 h. The cells were then washed twice with PBS at pH 7.4. To remove surface-bound activity, the cells were incubated for 10 min in glycine-HCl (1 M in PBS, pH 2.2) at room temperature. After washing twice with ice-cold PBS, the cells were lysed in 1.4 mL of lysis buffer (0.3 M NaOH, 0.2% SDS), and the internalized fraction was measured. For cells incubated at 4 °C, all washing and elution steps were performed using ice-cold buffer. Radioactivity was measured using a gamma counter (Packard Cobra II), normalized to 1 mio cells, and calculated as a percentage of the applied dose (% AD; see Figure 47).

[0281] Enzyme inhibition assay To measure the potential inhibitory effect of FAPI-04 on enzymatic FAP activity, an enzyme inhibition assay was performed using recombinant human FAP protein (1 pmol / well) in a 48-well plate. After incubating human FAP with FAPI-04 or talabostat (0–1000 nM / well) at 37°C for 30 min, the fluorescent FAP substrate Z-GP-AMC was added to a final concentration of 0–200 μM / well and incubated at 37°C for 60 min. The enzymatic activity of FAP was determined by measuring the fluorescence intensity of the reaction product, AMC, at 360 / 460 nm using a SpectraMax M2 plate reader (Molecular Devices, San Jose, USA) (see Figure 46).

[0282] Multiple administration of FAPI-04 to HT-1080-FAP tumor-bearing mice In the biodistribution experiments, 8-week-old BALB / c nu / nu mice (Children's River) were subcutaneously inoculated with 5 ml of HT-1080-FAP cells in the right trunk. Tumors were approximately 1 cm in size. 3 Once the rats reached 100 mg / kg, the radiolabeled compound was injected via the tail vein. 177 Group 1 received a single dose of Lu-FAPI-04 (2 MBq per animal), while Group 2 received two 1 MBq doses, with the second dose administered 4 hours after the first injection. Group 3 received a total of three doses: an initial dose of 1 MBq per mouse, followed by 0.5 MBq 2 hours later and an additional 0.5 MBq 4 hours later. Animals (n = 3 per time point) were sacrificed 8 and 24 hours after the initial injection. Distributed radioactivity was measured in all dissected organs and blood using a γ-counter (Cobra Autogamma, Packard). Values ​​are presented as a percentage of the administered dose per gram of tissue (%ID / g) (see Figure 48).

[0283] Example 8: In vitro and in vivo characterization of FAPI Experimental manipulation and clinical evaluation All in vitro and in vivo experiments and clinical evaluations of FAPI derivatives were performed as originally described in Loktev et al.1 and Lindner et al. 2 I followed the instructions.

[0284] result In vitro characterization of F-18-FAPI derivatives All experiments were performed in a similar manner with FAPI-42 (AlF-18 labeled) or FAPI-72 (F-18 nicotinamide labeled). [Table 11] Table 11: EC of selected FAPI derivatives measured by competitive binding assay 50 value

[0285] Measurement of blood pool clearance To estimate compound clearance rates, we calculated half-lives from cardiac SUVmean values ​​(0.375-60 min) as a representation of the blood pool by an assumed biphasic exponential decay. All selected compounds were cleared very rapidly, with half-lives of less than 10 min. The calculated plateau values ​​were higher for Ga-68-labeled FAPI-13, -21, and -36 and AlF-18-labeled FAPI-74, which theoretically corresponds to a higher fraction of compound not cleared due to nonspecific binding or remaining in the circulation (Table 12). As an example of fast clearance, the time-radioactivity curves for FAPI-04 and -46 between 0 and 15 min are shown in Figure 53. [Table 12] Table 12. Blood pool half-lives and hypothetical plateau values ​​of selected FAPI derivatives calculated from SUVmean values ​​by estimated two-phase exponential decay. For clarity, only rate-limiting half-life values ​​are listed.

[0286] Small animal imaging of F-18-FAPI derivatives in tumor-bearing mice Based on these findings, small-animal PET imaging was performed using F-18-labeled NOTA- and F-18-nicotinamide-labeled FAPI derivatives up to 140 min after radiotracer administration into HT-1080-FAP tumor-bearing mice. The F-18-nicotinamide derivatives, FAPI-72, -73, and -77, showed unfavorable hepatic accumulation and biliary excretion, whereas FAPI-78 was excreted via the kidney but showed no tumor uptake. In the case of the AlF-18-labeled NOTA derivatives, FAPI-74 and -75, high target specificity and fast clearance were observed, resulting in high-contrast images that enabled excellent visualization of FAP-positive tumors (Figure 50).

[0287] Organ distribution of F-18-FAPI derivatives in tumor-bearing mice To analyze the pharmacokinetic profile and in vivo tumor uptake, AlF-18-labeled FAPI-75 was intravenously administered to HT-1080-FAP tumor-bearing mice. The organ distribution of the radiolabeled compound was measured ex vivo in blood, healthy tissue, and tumor. As shown in Figure 51, this compound exhibited high tumor uptake, but higher accumulation in healthy tissue was observed compared to the Ga-68-labeled DOTA derivative, and its performance in PET imaging was comparable.

[0288] item The following items represent preferred embodiments of the present invention. 1. Formula (I): [ka] wherein Q, R, U, V, W, Y, and Z are independently present or absent, provided that at least three of Q, R, U, V, W, Y, and Z are present; Q, R, U, V, W, Y and Z are independently O, CH2, NR 4 , C=O, C=S, C=NR 4 , HCR 4 and R 4 CR 4 with the proviso that no two O's are immediately adjacent to each other; R 1 and R 2 are independently -H, -OH, halo, C 1-6 -Alkyl, -OC 1-6 -Alkyl, SC 1-6 - selected from the group consisting of alkyl; R 3 is selected from the group consisting of -H, -CN, -B(OH)2, -C(O)-alkyl, -C(O)-aryl, -C=CC(O)-aryl, -C=CS(O)2-aryl, -CO2H, -SO3H, -SON2NH2, -PO3H2 and 5-tetrazolyl; R 4 -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each -C 1-6 -alkyl is optionally substituted with 1 to 3 substituents selected from -OH, oxo and halo, and may be attached to Q, R, U, V, W, Y or Z;

[0289] R 5 -H, halo and C 1-6 - selected from the group consisting of alkyl; R 6 and R 7 independently -H, [ka] where R 6 and R 7 is not simultaneously H; where L is a linker; D, A, E and B are independently present or absent, preferably at least A, E and B are present, where, if present: D is a linker; A is NR 4, O, S, and CH2; E is C 1-6 -alkyl, [ka] selected from the group consisting of: where i is 1, 2, or 3; where j is 1, 2, or 3; where k is 1, 2, or 3; where m is 1, 2 or 3;

[0290] A and E together form a group selected from cycloalkyl, heterocycloalkyl, aryl and heteroaryl, preferably heterocycloalkyl, where A and E may be monocyclic, bicyclic and polycyclic, preferably monocyclic; each A and E may be -H, -C 1-6 -Alkyl, -OC 1-6 -Alkyl, -SC 1-6 -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C 1-6 -aralkyl, each of which is optionally substituted by 1 to 4 substituents selected from -C 1-6 -Alkyl may be substituted by 1 to 3 substituents selected from -OH, oxo and halo; and A, B, D, E or [ka] may be bonded to; B, S, NR 4 , N.R. 4 -O, NR 4 -C 1-6 -Alkyl, NR 4 -C 1-6 -Alkyl-NR 4and 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, the heterocycles preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably NR 4 -C 1-6 -Alkyl-NR 4 and the N-containing heterocycle is C 1-6 -Alkyl, aryl, C 1-6 -substituted by 1 to 3 substituents selected from the group consisting of -aralkyl; and R 8 is selected from the group consisting of a radioactive moiety, a chelating agent, a fluorescent dye, an imaging agent, and combinations thereof; [ka] is a 1-naphthyl moiety or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle has two ring atoms between the N atom and X; and the heterocycle may further contain one, two, or three heteroatoms selected from O, N, and S; and X is a C atom. or a pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof.

[0291] 2. (i) Q, R, and U are CH2 and independently present or absent; V is CH2, C=O, C=S or C=NR 4 and; W is NR 4 and; Y is HCR 4 and Z is C=O, C=S or C=NR 4 and / or (ii) Q and R are absent; U is CH2, present or absent; R 1 and R 2is independently selected from the group consisting of —H and halo; R 3 is selected from the group consisting of -H, -CN, and -B(OH)2; R 4 But -H and -C 1-6 -alkyl, wherein -C 1-6 The compound according to item 1, wherein -alkyl may be substituted by 1 to 3 substituents selected from -OH.

[0292] 3. [ka] but, [ka] 3. The compound of claim 1, wherein the compound is selected from the group consisting of:

[0293] 4. [ka] but [ka] The compound according to any one of items 1 to 3, selected from the group consisting of:

[0294] 5.R 5 and R 6 is H; R 7 but, [ka] where: D is absent; A is O; E is C 1-6 -alkyl or [ka] where m is 1, 2 or 3; A and E together [ka] forming a group selected from: B is NR 4 -C 1-6 -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, the heterocycle preferably further containing one or two heteroatoms selected from O, N and S, preferably further containing one or two nitrogen atoms, wherein preferably the N-containing heterocycle is C 1-6 -Alkyl, aryl and C 1-6 - the compound according to any one of items 1 to 4, which is substituted by 1 to 3 substituents selected from the group consisting of aryl, ...

[0295] 6. (i) The N-containing heterocycle contained in B is of the formula: [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may further contain one nitrogen; [ka] is attached to position 1, 2 or 3, preferably to position 2; l is 1 or 2] and / or (ii) the N-containing heterocycle contained in B is [ka] wherein the N-containing heterocycle is selected from the group consisting of C 1-6 -substituted by alkyl; Here, the N-containing heterocycle contained in B is [ka] wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may further contain one nitrogen, and may contain one or more side chains (e.g. derived from amino acids); [ka] is attached to position 1, 2 or 3, preferably to position 2; o is 1 or 2, Preferably, the N-containing heterocycle contained in B is [ka] When B is [ka] and more preferably, the N-containing heterocycle contained in B is [ka] When B is [ka] 6. The compound according to any one of items 1 to 5, wherein

[0296] 7. Q, R and U are absent; V is C=O; W is NH; Y is CH2; Z is C=O; R 1 and R 2 is independently selected from the group consisting of —H and halo; R 3 is -CN; R 5 and R 6 is H; R 7 but [ka] where: D is absent; A is O, S, CH2, NH or NCH3; E is C 1-6 -alkyl or [ka] where m is 1, 2, or 3; or A and E together [ka] forming a group selected from: B is NH-C 1-6 -alkyl, [ka] and B is C 1-3 optionally substituted by alkyl; and [ka] teeth [ka] 7. The compound according to any one of items 1 to 6, wherein

[0297] 8.C 1-6 alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl, and / or C 1-6 8. The compound according to any one of items 1 to 7, wherein aralkyl is selected from the group consisting of benzyl, phenyl-ethyl, phenyl-propyl and phenyl-butyl.

[0298] 9.R 8is a radioactive moiety which is a fluorescent isotope, a radioisotope, a radiopharmaceutical or a combination thereof, preferably an α-radiation emitting isotope, a β-radiation emitting isotope, a γ-radiation emitting isotope, an Auger electron emitting isotope, an X-ray emitting isotope, a fluorescent emitting isotope, e.g. 11 C. 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Ag, preferably 18 F, 64 Cu, 68 Ga,90 Y, 99m Tc, 153 Sm, 177 Lu and 188 9. The compound according to any one of items 1 to 8, wherein the radioactive moiety is selected from the group consisting of Re.

[0299] 10.R 8 is selected from the group consisting of the following types of fluorescent dyes: xanthenes, acridines, oxazines, cyanines, styryl dyes, coumarins, porphines, metal-ligand complexes, fluorescent proteins, nanocrystals, perylenes, boron-dipyrromethenes and phthalocyanines, as well as composites and combinations of these types of dyes.

[0300] 11.R 8 is a chelating agent in the form of a complex with a divalent or trivalent metal cation, preferably selected from the group consisting of 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), triethylenetetramine (TETA), iminodiacetic acid, diethylenetriamine-N,N,N',N',N''-pentaacetic acid (DTPA), bis-(carboxymethylimidazole)glycine and 6-hydrazinopyridine-3-carboxylic acid (HYNIC).

[0301] 12.R 8 9. The compound according to any one of items 1 to 8, wherein the compound is a contrast agent comprising or consisting of a paramagnetic agent, preferably wherein the paramagnetic agent comprises or consists of paramagnetic nanoparticles.

[0302] 13. A pharmaceutical composition comprising or consisting of at least one compound according to items 1 to 12, and optionally a pharmaceutically acceptable carrier and / or excipient.

[0303] 14. A compound according to any one of items 1 to 12 or a pharmaceutical composition according to item 13 for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activating protein (FAP) in an animal or human subject, preferably wherein the disease characterized by overexpression of fibroblast activating protein (FAP) is selected from the group consisting of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and keloid disorders, preferably wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, cholangiocarcinoma, renal clear cell carcinoma, neuroendocrine tumor, tumor-induced osteomalacia, sarcoma, CUP (cancer of unknown primary stage), thymic carcinoma, desmoid tumor, glioma, astrocytoma, cervical cancer and prostate cancer.

[0304] 15. A kit comprising or consisting of the compound according to any one of items 1 to 12 or the pharmaceutical composition according to item 13 and instructions for diagnosing a disease.

Claims

1. Formula (I): 【Chemistry 1】 wherein Q, R, U, V, W, Y, and Z are independently present or absent, provided that at least three of Q, R, U, V, W, Y, and Z are present; Q, R, U, V, W, Y and Z are independently O, CH ２ , N.R. ４ , C=O, C=S, C=NR ４ , HCR ４ and R ４ CR ４ with the proviso that no two O's are immediately adjacent to each other; R １ and R ２ are independently —H, —OH, halo, C １－６ -alkyl, -O-C １－６ -Alkyl, S—C １－６ - selected from the group consisting of alkyl; R ３ -H, -CN, -B(OH) ２ , —C(O)-alkyl, —C(O)-aryl, —C═C—C(O)-aryl, —C═C—S(O) ２ -aryl, -CO ２ H, -SO ３ H, -SO ２ NH ２ , -PO ３ H ２ and 5-tetrazolyl; R ４ is -H, -C １－６ -alkyl, -O-C １－６ -Alkyl, -S-C １－６ -alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl and -C １－６ -aralkyl, each -C １－６ -alkyl is optionally substituted with 1 to 3 substituents selected from -OH, oxo and halo and may be attached to Q, R, U, V, W, Y or Z; R ５ is -H, halo and C １－６ - selected from the group consisting of alkyl; R ６ and R ７ are independently -H, 【Chemistry 2】 where R ６ and R ７ is not simultaneously H; where L is a linker; D, A, E and B are independently present or absent, preferably at least A, E and B are present, where, if present: D is a linker; A is NR ４ , O, S and CH ２ selected from the group consisting of: E is C １－６ - alkyl, 【Transformation 3】 selected from the group consisting of: where i is 1, 2 or 3; where j is 1, 2 or 3; where k is 1, 2 or 3; wherein m is 1, 2, or 3; B is S, NR ４ , N.R. ４ -O, NR ４ -C １－６ -Alkyl, NR ４ -C １－６ -Alkyl-NR ４ and 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycles, which heterocycles preferably further contain one or two heteroatoms selected from O, N and S, and preferably further contain one or two nitrogen atoms, wherein preferably NR ４ -C １－６ -Alkyl-NR ４ and the N-containing heterocycle is C １－６ -Alkyl, aryl, C １－６ -substituted by 1 to 3 substituents selected from the group consisting of aralkyl; and R ８ is selected from the group consisting of a radioactive moiety, a chelator, a fluorescent dye, an imaging agent, and combinations thereof; 【Chemistry 4】 is a 1-naphthyl moiety or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, wherein the heterocycle has two ring atoms between the N atom and X; and the heterocycle may further contain one, two, or three heteroatoms selected from O, N, and S; and X is a C atom. or a pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof.

2. (i) Q, R and U are CH ２ and is independently present or absent; V is CH ２ , C═O, C═S or C═NR ４ and W is NR ４ and Y is HCR ４ and Z is C═O, C═S or C═NR ４ and / or (ii) Q and R are absent; U is CH ２ is present or absent; R １ and R ２ is independently selected from the group consisting of —H and halo; R ３ -H, -CN and -B(OH) ２ selected from the group consisting of: R ４ is -H and -C １－６ -alkyl, wherein -C １－６ The compound of claim 1, wherein -alkyl is optionally substituted with 1 to 3 substituents selected from -OH. 【Request Item 3】 【Chemistry 5】 but, 【Transformation 6】 3. The compound of claim 1, wherein the compound is selected from the group consisting of: 【Request Item 4】 【Chemistry 7】 but, 【Transformation 8】 The compound according to any one of claims 1 to 3, selected from the group consisting of:

5. R ５ and R ６ is H; R ７ but, 【Chemistry 9】 where: D is absent; A is O; E is C １－６ - alkyl or 【Chemistry 10】 where m is 1, 2 or 3; B is NR ４ -C １－６ -alkyl or a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, which heterocycle preferably further contains one or two heteroatoms selected from O, N and S, preferably further contains one or two nitrogen atoms, wherein preferably the N-containing heterocycle is C １－６ - alkyl, aryl and C １－６ - aralkyl.

6. (i) the N-containing heterocycle contained in B is of the formula: 【Chemistry 11】 wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may further contain one nitrogen; 【Chemistry 12】 is attached to position 1, 2 or 3, preferably to position 2; l is 1 or 2. and / or an aromatic or non-aromatic monocyclic heterocycle of the formula (ii) the N-containing heterocycle contained in B is 【Chemistry 13】 is selected from the group consisting of wherein the N-containing heterocycle contained in B is 【Chemistry 14】 wherein the heterocycle may further contain one or two heteroatoms selected from O, N and S, and may further contain one nitrogen, and may also contain one or more side chains (e.g. derived from amino acids); 【Chemistry 15】 is attached to position 1, 2 or 3, preferably to position 2; o is 1 or 2; Preferably, the N-containing heterocycle contained in B is 【Chemistry 16】 When B is 【Chemistry 17】 and more preferably, the N-containing heterocycle contained in B is [Chemistry 18] When B is 【Chemistry 19】 The compound according to any one of claims 1 to 5,

7. Q, R and U are absent; V is C=O; W is NH; Y is CH ２ and Z is C=O; R １ and R ２ is independently selected from the group consisting of —H and halo; R ３ is -CN; R ５ and R ６ is H; R ７ but 【Chemistry 20】 where: D is absent; A is O; E is C １－６ - alkyl or 【Chemistry 21】 where m is 1, 2 or 3; B is NH-C １－６ - alkyl, 【Chemistry 22】 and 【Chemistry 23】 teeth 【Chemistry 24】 The compound according to any one of claims 1 to 6,

8. C １－６ alkyl is selected from the group consisting of methyl, ethyl, propyl, i-propyl, butyl, sec-butyl, tert-butyl, pentyl and hexyl, and / or C １－６ A compound according to any one of claims 1 to 7, wherein aralkyl is selected from the group consisting of benzyl, phenyl-ethyl, phenyl-propyl and phenyl-butyl.

9. R ８ is a radioactive moiety which is a fluorescent isotope, a radioisotope, a radiopharmaceutical or a combination thereof, preferably an α-radiation emitting isotope, a β-radiation emitting isotope, a γ-radiation emitting isotope, an Auger electron emitting isotope, an X-ray emitting isotope, a fluorescent emitting isotope, e.g. １８ F. ５１ Cr, ６７ Ga, ６８ Ga, １１１ In, ９９ｍ Tc, １８６ Re, １８８ Re, １３９ La, １４０ La, １７５ Yb, １５３ Sm, １６６ Ho, ８８ Y. ９０ Y. １４９ Pm, １６５ Dy, １６９ Er, １７７ Lu, ４７ Sc, １４２ Pr, １５９ Gd, ２１２ Bi, ２１３ Bi, ７２ As, ７２ Se, ９７ Ru, １０９ Pd, １０５ Rh, １０１ｍ Rh, １１９ Sb, １２８ Ba, １２３ I, １２４ I, １３１ I, １９７ Hg, ２１１ At, １５１ EU, １５３ EU, １６９ EU, ２０１ Tl, ２０３ Pb, ２１２ Pb, ６４ Cu, ６７ Cu, １８８ Re, １８６ Re, １９８ Au, ２２５ Ac, ２２７ Th and １９９ The compound of any one of claims 1 to 8, wherein the radioactive moiety is selected from the group consisting of Ag.

10. R ８ is selected from the group consisting of the following types of fluorescent dyes: xanthenes, acridines, oxazines, cyanines, styryl dyes, coumarins, porphines, metal-ligand complexes, fluorescent proteins, nanocrystals, perylenes, boron-dipyrromethenes and phthalocyanines, as well as composites and combinations of these types of dyes.

11. R ８ is a chelating agent in the form of a complex with a divalent or trivalent metal cation, preferably selected from the group consisting of 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), triethylenetetramine (TETA), iminodiacetic acid, diethylenetriamine-N,N,N',N',N''-pentaacetic acid (DTPA), bis-(carboxymethylimidazole)glycine and 6-hydrazinopyridine-3-carboxylic acid (HYNIC).

12. R ８ is a contrast agent comprising or consisting of a paramagnetic agent, preferably wherein the paramagnetic agent comprises or consists of paramagnetic nanoparticles.

13. A pharmaceutical composition comprising or consisting of at least one compound according to claims 1 to 12, and optionally a pharmaceutically acceptable carrier and / or excipient.

14. 14. A compound according to any one of claims 1 to 12 or a pharmaceutical composition according to claim 13 for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in an animal or human subject, preferably wherein the disease characterized by overexpression of fibroblast activation protein (FAP) is selected from the group consisting of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and keloid disorders, preferably wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, cholangiocarcinoma, renal clear cell carcinoma, neuroendocrine tumors, tumor-induced osteomalacia, sarcoma, CUP (cancer of unknown primary stage), thymic carcinoma, desmoid tumor, glioma, astrocytoma, cervical cancer and prostate cancer.

15. A kit comprising or consisting of a compound according to any one of claims 1 to 12 or a pharmaceutical composition according to claim 13 and instructions for the diagnosis of a disease.

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