Compounds comprising fibroblast activation protein ligands and uses thereof

Abstract

To provide compounds, including cyclic peptides and chelators, suitable as diagnostic and / or therapeutic agents, especially when conjugated to a diagnostically and / or therapeutically active effector, and uses thereof.SOLUTION: A compound selected from the group consisting of compound Hex - [Cys (tMeBn (DOTA-APE)) - Pro-Pro-Thr-Gln-Phe-Cys] - OOH (3Bp-3545), and compound Hex - [Cys (tMeBn (DOTA-AP)) - Pro-Pro-Thr-Gln-Phe-Cys] - Asp-NH2 (3Bp-3400) is provided.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to compounds; inhibitors of fibroblast activation proteins (FAPs); compositions comprising the compounds and inhibitors, respectively; compounds, inhibitors, and compositions, respectively, for use in methods for the diagnosis of disease; compounds, inhibitors, and compositions, respectively, for use in methods for the treatment of disease; compounds, inhibitors, and compositions, respectively, for use in methods for the diagnosis and treatment of disease, also referred to as "thera(g)nosis" or "thera(g)nostics"; compounds, inhibitors, and compositions, respectively, for use in methods for delivering effectors to FAP-expressing tissue; methods for the diagnosis of disease using the compounds, inhibitors, and compositions, respectively; methods for the treatment of disease using the compounds, inhibitors, and compositions, respectively; methods for the diagnosis and treatment of disease, also referred to as "thera(g)nosis" or "thera(g)nostics" using the compounds, inhibitors, and compositions, respectively; and methods for the delivery of effectors to FAP-expressing tissue using the compounds, inhibitors, and compositions, respectively. [Background technology]

[0002] Despite the increasing availability of therapeutic options, cancer remains the second leading cause of death worldwide. Treatment strategies primarily focus on targeting malignant cancer cells themselves, ignoring the ever-present surrounding tumor microenvironment (TME), which limits the access of therapeutic cancer cell agents (Valkenburg et al., Nat Rev Clin Oncol, 2018, 15:366). The TME is part of the tumor mass and consists of not only a heterogeneous population of cancer cells but also various resident and infiltrating host cells, secreted factors, and extracellular matrix proteins (Quail et al., Nat Med, 2013, 19:1423). The predominant cell type found in the TME is cancer-associated fibroblasts (CAFs) (Kalluri, Nat Rev Cancer, 2016, 16:582). Many different cell types have been described as sources and origins for CAFs, such as fibroblasts, mesenchymal stem cells, smooth muscle cells, cells of epithelial origin, or endothelial cells (Madar et al., Trends Mol Med, 2013, 19:447). CAFs exhibit mesenchymal-like characteristics and are often the predominant cell type within solid tumor masses. CAFs have attracted increasing interest as players in tumor progression and homeostasis (Gascard et al., Genes Dev, 2016, 30:1002; LeBleu et al., Dis Model Mech, 2018, 11).

[0003] In recent years, fibroblast activation protein (FAP) has lost its reputation as a marker of CAFs (Shiga et al., Cancers (Basel), 2015, 7:2443; Pure et al., Oncogene, 2018, 37:4343; Jacob et al., Curr Mol Med, 2012, 12:1220). Due to the ubiquitous presence of CAFs in tumors and stroma, FAP has been identified as a suitable marker for radiopharmaceutical diagnosis and a suitable target for radiopharmaceutical therapy (Siveke, J Nucl Med, 2018, 59:1412).

[0004] Fibroblast activation protein alpha (FAP) is a type II transmembrane serine protease and a member of the S9 prolyl oligopeptidase family (Park et al., J Biol Chem, 1999, 274:36505). The closest family member, DPP4, shares 53% homology with FAP. Like other DPP enzymes (DPP4, DPP7, DPP8, and DPP9), FAP possesses post-proline exopeptidase activity. Furthermore, FAP possesses endopeptidase activity, similar to prolyl oligopeptidase / endopeptidase (POP / PREP). The FAP gene is highly conserved among various species. The extracellular domain of human FAP shares 90% amino acid sequence identity with mouse and rat FAP. Mouse FAP shares 97% sequence identity with rat FAP.

[0005] Structurally, FAP is a 760-amino acid transmembrane protein composed of a short N-terminal cytoplasmic tail (6 amino acids), a single transmembrane domain (20 amino acids), and a 734-amino acid extracellular domain (Aertgeerts et al., J Biol Chem, 2005, 280:19441). This extracellular domain consists of an eight-bladed β-propeller domain and an α / β-hydrolase domain. The catalytic triad, consisting of Ser624, Asp702, and His734, is located at the interface between the β-propeller and hydrolase domains. The active site is accessible through the central hole of the β-propeller domain or through a narrow cavity between the β-propeller and hydrolase domains. FAP is not active as a monomer, but forms active homodimers and heterodimers with DPP4 (Ghersi et al., Cancer Res, 2006, 66:4652). Soluble homodimeric FAPs have also been described (Keane et al., FEBS Open Bio, 2013, 4:43; Lee et al., Blood, 2006, 107:1397).

[0006] FAP has dual enzymatic activity (Hamson et al., Proteomics Clin Appl, 2014, 8:454). Its dipeptidyl peptidase activity cleaves two amino acids N-terminal to the proline residue. FAP substrates rapidly cleaved by its dipeptidyl peptidase activity include neuropeptide Y, peptide YY, substance P, and B-type natriuretic peptide. Collagens I and III, FGF21, and α2-antiplasmin have been shown to be cleaved by FAP's endopeptidase activity. Although FAP cannot cleave native collagens, predigestion with other proteases, such as matrix metalloproteinases, facilitates further collagen cleavage by FAP. Collagen processing can affect the migration ability of cancer cells. In addition to increasing cancer cell invasiveness through extracellular matrix remodeling, several other FAP-mediated tumor-promoting roles have been proposed, including increased proliferation and angiogenesis. Furthermore, stromal expression of FAP is associated with escape from immune surveillance in various cancers, suggesting a role in antitumor immunity (Pure et al., Oncogene, 2018, 37:4343).

[0007] FAP is transiently expressed during normal development but is only rarely expressed in healthy adult tissues. In transgenic mice, FAP has been shown to be expressed by adipose tissue, skeletal muscle, skin, bone, and pancreas (Pure et al., Oncogene, 2018, 37: 4343; Roberts et al., J Exp Med, 2013, 210: 1137). However, FAP knockout mice have a healthy phenotype, suggesting a redundant role under normal conditions (Niedermeyer et al., Mol Cell Biol, 2000, 20: 1089). At sites of active tissue remodeling, including wound healing, fibrosis, arthritis, atherosclerosis, and cancer, FAP becomes highly upregulated in stromal cells (Pure et al., Oncogene, 2018, 37: 4343).

[0008] FAP expression in the tumor stroma of 90% of epithelial cancers was first reported in 1990 using the monoclonal antibody F19 (Garin-Chesa et al., Proc Natl Acad Sci USA, 1990, 87:7235; Rettig et al., Cancer Res, 1993, 53:3327). FAP-expressing stromal cells were further characterized as cancer-associated fibroblasts (CAFs) and cancer-associated pericytes (Cremasco et al., Cancer Immunol Res, 2018, 6:1472). FAP expression on malignant epithelial cells has also been reported, although its significance remains to be determined (Pure et al., Oncogene, 2018, 37:4343). Busek et al. (Busek et al., Fron Table 1 below, taken from Biosci (Landmark Ed), 2018, 23:1933, summarizes the expression of FAP in various malignant tumors showing tumor type and cellular expression.

[0009] [Table 1-1]

[0010] [Table 1-2]

[0011] FAP expression in CAFs has been demonstrated in nearly all carcinomas and sarcomas (Pure et al., Oncogene, 2018, 37:4343; Busek et al., Front Biosci (Landmark Ed), 2018, 23:1933). Furthermore, CAFs are present in hematological malignancies (Raffaghello et al., Oncotarget, 2015, 6:2589). Therefore, the use of FAP as a therapeutic target is not limited to any particular tumor entity.

[0012] The abundance of FAP-expressing CAFs has been reported to correlate with poor prognosis. Across various human tumor indications, FAP expression has been reported to correlate with higher tumor grade and worse overall survival (Pure et al., Oncogene, 2018, 37:4343).

[0013] As mentioned above, FAP and FAP-expressing cells present in the tumor microenvironment are shown to significantly affect tumor progression (Hanahan et al., Cancer Cell, 2012, 21:309). Furthermore, due to their relatively selective expression in tumors, FAP is It is considered a suitable target for therapeutic and diagnostic agents as described below (Siveke, J Nucl Med, 2018, 59:1412; Christiansen et al., Neoplasia, 2013, 15:348; Zi et al., Mol Med Rep, 2015, 11:3203).

[0014] Shortly after their discovery, FAPs were exploited as therapeutic targets in cancer, and to date, various strategies have been explored, including, for example, inhibition of FAP enzymatic activity, elimination of FAP-positive cells, or targeted delivery of cytotoxic compounds.

[0015] In 2007, talabostat (Val-boro-Pro, PT-100), an inhibitor of FAP and DPP4, was developed by Point Therapeutics (e.g., U.S. Patent No. 6,890,904, described in WO9916864). Pennisi et al. (Pennisi et al., Br J Haematol, 2009, 145:775) observed a reduction in tumor growth in multiple myeloma animal models as well as in cancer syngeneic mouse models. In addition, several other prolylboronic acid derivatives have been developed, including F These derivatives have been reported as putative selective inhibitors of AP. These derivatives exhibit instability in aqueous environments at physiological pH (Coutts et al., J Med Chem, 1996, 39:2087) and nonspecific reactivity with other enzymes.

[0016] WO2008 / 116054 disclosed hexapeptide derivatives in which the compounds contain a C-terminal bisamino or boronic acid functionality. US2017 / 0066800 disclosed pseudopeptide inhibitors, such as M83, that are effective against FAP. These inhibitors were evaluated in lung and colon cancer xenografts in immunodeficient mice. Tumor growth inhibition was observed (Jackson et al., Neoplasia, 2015, 17:43). These pseudopeptides inhibit the activity of both prolyl oligopeptidase (POP / PREP) and FAP, thereby precluding their use as specific therapeutic FAP inhibitors.

[0017] US2008 / 280856 disclosed nanomolar concentrations of boronic acid-based inhibitors that exhibit dual specific inhibition of FAP and PREP, thereby precluding their use as specific therapeutic FAP inhibitors.

[0018] Cyclic peptide-based FAP inhibitors have been disclosed, for example, in WO2016 / 146174 and WO2006 / 042282. WO2016 / 146174 discloses a peptide for the diagnosis and treatment of FAP-expressing tumors that exhibits specificity for FAP, and the closely related homolog DPP4 is not recognized by the peptide. WO2006 / 042282 discloses a polypeptide for the treatment of melanoma. In nude mice, inhibition of melanoma growth and melanoma metastasis was demonstrated.

[0019] WO99 / 75151 and WO01 / 68708 disclosed a humanized FAP monoclonal antibody, F19 (sibrotuzumab). Furthermore, the anti-FAP antibody F19 and its humanized versions were disclosed in WO99 / 57151 and WO01 / 68708. Development approaches included, for example, the generation of high-affinity, species-cross-reactive, FAP-specific scFvs, which were converted into bivalent derivatives (Brocks et al., Mol Med, 2001, 7:461). In phase I and II clinical trials, sibrotuzumab demonstrated specific tumor enrichment but failed to demonstrate measurable therapeutic activity in patients with metastatic colorectal cancer, with only 2 of 17 patients achieving stable disease (Hofheinz et al., Onkologie, 2003, 26:44). The F19 antibody was shown not to block any cellular or protease functions of FAP, which may explain the lack of therapeutic efficacy (Hofheinz et al., Onkologie, 2003, 26:44; Scott et al., Clin Cancer Res, 2003, 9:1639).

[0020] US2018 / 022822 disclosed novel molecules that specifically bind to human FAP and its epitopes as human-derived antibodies and chimeric antigen receptors (CARs) useful in the treatment of diseases and conditions induced by FAP. Treatment of mice bearing orthotopic MC38 colorectal tumors with anti-FAP antibodies reduced tumor diameter and the number of metastases. WO2012 / 020006 disclosed glycoengineered antibodies carrying modified oligosaccharides in the Fc region. Subsequently, bispecific antibodies specific for FAP and DR5 were developed as the subject of WO2014 / 161845. These antibodies induced tumor cell apoptosis in in vitro and in vivo preclinical tumor models with FAP-positive stroma (Brunker et al., Mol Cancer Ther, 2016, 15:946). Antibody-drug conjugates and immunotoxins targeting FAP are described in WO2015 / 118030. In vitro toxicity as well as in vivo inhibition of tumor growth was demonstrated following application of the anti-hu / moFAP hu36:cytolysin ADC candidate. It is unclear whether these antibodies were able to inhibit FAP activity.

[0021] A small molecule FAP inhibitor based on (4-quinolinol)glycyl-2-cyanopyrrolidine, which exhibits low nanomolar inhibitory potency and high selectivity against related DPPs and PREPs, was described by Jansen et al. (Jansen et al., J Med Chem, 2014, 57:3053; Jansen et al., ACS Med Chem Lett, 2013, 4:491) and disclosed in WO2013 / 107820. However, that compound is not structurally related to the compounds of the present invention and contains a warhead that provides covalent binding to FAP.

[0022] Recently, several FAP-targeted radiopharmaceutical approaches have been developed, which are illustratively described herein. WO2010 / 036814 disclosed small molecule inhibitors of FAP for use as therapeutic agents by inhibition of FAP enzymatic activity or as radiopharmaceuticals by binding to FAP.

[0023] WO2019 / 083990 disclosed imaging and radiotherapeutic agents based on small molecule FAP inhibitors described by Jansen et al. (Jansen et al., J Med Chem, 2014, 57:3053; Jansen et al., ACS Med Chem Lett, 2013, 4:491). Furthermore, several authors have reported that Jansen et al. (Jansen et al., J Med Chem, 2014, 57:3053; Jansen et al., ACS Med Chem Lett, 2013, 4:491) have described the selective uptake of FAP inhibitor-based imaging and radiotherapeutic agents in tumors of cancer patients (Lindner et al., J Nucl Med, 2018, 59:1415; Loktev, et al., J Nucl Med, 2018, 59:1423; Giesel et al., J Nucl Med, 2019, 60:386; Loktev et al., J Nucl Med, 2019, Mar 8 (electronic version ahead of print); Giesel et al., Eur J Nucl Med Mol Imaging, 2019, 46:1754; Kratochwil et al., J Nucl Med, 2019, 60:801).

[0024] 131 Clinical evaluation of an I-labeled, humanized form of the F19 antibody (sibrotuzumab) demonstrated selective uptake by tumors, but not normal tissues, in patients with colorectal cancer or non-small cell lung cancer (Scott, et al., Clin Cancer Res, 2003, 9:1639). This may be due to the long circulation time of the antibody, making it unsuitable for diagnostic, therapeutic, or diagnostic-therapeutic procedures involving radionuclides.

[0025] WO2011 / 040972 disclosed a high-affinity antibody that recognizes both human and mouse FAP antigens as a potent radioimmunoconjugate. ESC11 IgG1 induces down-modulation and internalization of surface FAP (Fischer et al., Clin Cancer Res, 2012, 18:6208). WO2017 / 211809 disclosed a tissue-targeted thorium-227 conjugate in which the targeting moiety has specificity for FAP. However, the long circulation time of antibodies makes them unsuitable for diagnostic, therapeutic, or diagnostic-therapeutic procedures involving radionuclides.

[0026] FAP has also been described to be involved in diseases other than oncological indications, examples of which are given below. Fibroblast-like synoviocytes in rheumatoid arthritis joints of patients show significantly increased expression of FAP (Bauer et al., Arthritis Res Ther, 2006, 8:R171; Milner et al., Arthritis Res Ther, 2006, 8:R23). In rheumatoid arthritis, stromal cells are important in organizing the structure of the synovial tissue of the joint by producing extracellular matrix components, recruiting infiltrating immune cells, and secreting inflammatory mediators. They play a crucial role. There is considerable evidence supporting the role of these cells in driving the persistence of inflammation and joint damage (Bartok et al., Immunol Rev, 2010, 233:233; Turner et al., Curr Opin Rheumatol, 2015, 27:175). In rheumatoid arthritis, FAP plays a pathological role in cartilage turnover, at least by promoting proteoglycan loss and subsequent cartilage degradation (Bauer et al., Arthritis Res Ther, 2006, 8:R171; Waldele et al., Arthritis Res Ther, 2015, 17:12). Therefore, it can serve as a marker for patient stratification, or as a therapeutic target for the evaluation and follow-up of treatment success (Bauer et al., Arthritis Res Ther, 2006, 8:R171). In mice, treatment response is 99m This has been demonstrated using SPECT / CT imaging with Tc-labeled anti-FAP antibodies (van der Geest et al., Rheumatology (Oxford), 2018, 57:737; Laverman et al., J Nucl Med, 2015, 56:778; van der Geest et al., J Nucl Med, 2017, 58:151).

[0027] Furthermore, FAP has been recognized not only as a marker of activated fibroblasts in the injury response (Tillmanns et al., Int J Cardiol, 2013, 168:3926) but also as an important player in the wound healing process (Ramirez-Montagut et al., Oncogene, 2004, 23:5435). Jing et al. demonstrated a time-dependent course of changes in FAP expression after burn wound injury in rats (Jing et al., Nan Fang Yi Ke Da Xue Xu Bao, 2013, 33:615). Inhibition of FAP activity in reactive wound fibroblasts in keloid scars, a common benign fibroproliferative reticular skin lesion, may provide a therapeutic option for preventing disease progression (Dienus et al., Arch Dermatol Res, 2010, 302:725).

[0028] In fibrosis, for example, in idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis, upregulated expression of FAP has been observed. In an ex vivo model of Crohn's disease, a chronic inflammatory bowel disease characterized by excessive and unbalanced extracellular matrix (ECM) deposition, upregulated FAP expression was observed. FAP inhibition reconstituted extracellular matrix homeostasis (Truffi et al., Inflamm Bowel Dis, 2018, 24:332). A similar observation was made by Egger et al. (Egger et al., Eur J Pharmacol, 2017, 809:64) using a mouse model of pulmonary fibrosis. Inhibition of FAP results in a reduction of fibrotic pathology. FAP is also expressed in areas of tissue remodeling in chronically injured livers (Wang et al., Front Biosci, 2008, 13:3168), and FAP expression by hepatic stellate cells correlates with the histological severity of liver disease (Gorrell et al., Adv Exp Med Biol, 2003, 524:235). Therefore, FAP is also a promising target in the treatment of liver fibrosis (Lay et al., Front Biosci(Landmark Ed),2019,24:1).

[0029] FAP is expressed in atherosclerotic lesions and is upregulated in activated vascular smooth muscle cells (Monslow et al., Circulation, 2013, 128:A17597). Monslow et al. demonstrated that targeted inhibition of FAP in atherosclerotic lesions can reduce overall lesion volume, inhibit inflammatory cell homing, and increase lesion stability through its ability to alter lesion structure by favoring matrix-rich lesions over inflammation. More importantly, many atherosclerotic pathologies share a common pathological feature: the rupture of atherosclerotic plaques, which leads to atherosclerotic lesions (Davies et al., Br Heart J, 1985, 53:363; Falk, Am J Cardiol, 1989, 63:114e). Rupture of the fibrous cap in advanced atherosclerotic plaques is an important precipitating factor for acute coronary syndromes, which can result in myocardial infarction and sudden cardiac death. Key events in promoting plaque instability One of the mechanisms is the degradation of the fibrous cap, which exposes the underlying thrombogenic plaque core to blood flow, leading to thrombosis and subsequent vascular occlusion (Farb et al., Circulation, 1996, 93:1354; Virmani et al., J Am Coll Cardiol, 2006, 47:C13). Brokopp et al. showed that FAP contributes to type I collagen breakdown in the fibrous cap (Brokopp et al., Eur Heart J, 2011, 32:2713). Radiolabeled tracers have been developed and their applicability for atherosclerosis imaging has been demonstrated (Meletta et al., Molecules, 2015, 20:2081). Summary of the Invention [Problem to be solved by the invention]

[0030] The problem underlying the present invention is to provide compounds suitable as diagnostic and / or therapeutic agents, particularly when conjugated to a diagnostically and / or therapeutically active effector. A further problem underlying the present invention is to provide compounds suitable as diagnostic and / or therapeutic agents, particularly when conjugated to a diagnostically and / or therapeutically active effector, such that the compounds are potent inhibitors of FAP activity, preferably with a pIC50 of greater than or equal to 6.0. A further problem underlying the present invention is to provide compounds suitable as diagnostic and / or therapeutic agents in the diagnosis and / or treatment of diseases in which diseased cells and / or diseased tissues express FAP, particularly when conjugated to a diagnostically and / or therapeutically active effector. A further problem underlying the present invention is to provide compounds suitable for delivering diagnostically and / or therapeutically effective agents to diseased cells and / or diseased tissues, respectively, more particularly to diseased cells and / or diseased tissues expressing FAP, preferably diseased tissues that include or contain cancer-associated fibroblasts. The present invention also provides methods for diagnosing, treating, and / or preventing diseases, as well as methods for the combined diagnosis and treatment of diseases, preferably diseases involving cells and / or tissues expressing FAP, more particularly diseased cells and / or tissues expressing FAP, and preferably diseased tissues including or containing cancer-associated fibroblasts. A further object of the present invention is to provide methods for identifying subjects likely to respond or not respond to disease treatment, and methods for selecting subjects likely to respond or not respond to disease treatment from a group of subjects. A further object of the present invention is to provide pharmaceutical compositions comprising compounds having the characteristics outlined above. A further object of the present invention is to provide kits suitable for use in any of the above methods.

[0031] There is a need for compounds that are suitable as diagnostic and / or therapeutic agents, particularly when conjugated to a diagnostically and / or therapeutically active effector. There is also a need for compounds that are suitable as diagnostic and / or therapeutic agents, particularly when conjugated to a diagnostically and / or therapeutically active effector, such that the compound is a potent inhibitor of FAP activity, preferably where the compound has a pIC50 of equal to or greater than 6.0. There is also a need for compounds that are suitable as diagnostic and / or therapeutic agents in the diagnosis and / or treatment of diseases in which diseased cells and / or diseased tissues express FAP, particularly when conjugated to a diagnostically and / or therapeutically active effector. There is also a need for compounds that are suitable for delivering diagnostically and / or therapeutically effective agents to diseased cells and / or diseased tissues, respectively, more particularly to diseased cells and / or diseased tissues that express FAP, preferably where the diseased tissues include or contain cancer-associated fibroblasts. Also provided are methods for the diagnosis of diseases, methods for the treatment and / or prevention of diseases, and methods for the combined diagnosis and treatment of diseases, preferably such

[0004] There is a need for a method wherein the disease involves cells and / or tissues expressing a FAP, more particularly diseased cells and / or tissues expressing a FAP, and preferably the diseased tissues include or contain cancer-associated fibroblasts. Further, there is a need for a method for identifying subjects likely to respond or not respond to disease treatment, and for selecting subjects likely to respond or not respond to disease treatment from a group of subjects. Further, there is a need for a pharmaceutical composition comprising a compound having the characteristics outlined above. Furthermore, there is a need for a kit suitable for use in any of the above methods. The present invention satisfies these needs. [Means for solving the problem]

[0032] These and other problems are solved by the subject matter of the appended claims. These and other problems underlying the present invention are also solved by the following embodiments. Embodiment 1. A compound of the formula:

[0033] [ka]

[0034] the compound Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554), and the compound of the formula

[0035] [ka]

[0036] The compound Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr -Gln-Phe-Cys]-Asp-NH2 (3BP-3407) A compound selected from the group consisting of: Embodiment 2. The following formula:

[0037] [ka]

[0038] 2. The compound of embodiment 1, wherein the compound is Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554). Embodiment 3. The following formula:

[0039] [ka]

[0040] 2. The compound according to embodiment 1, which is the compound Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407) of formula: Embodiment 4. The compound of any one of embodiments 1 to 3, wherein any S atom that can be oxidized, preferably an S atom of a thioether group, is present as -S-, -S(O)-, or -S(O2)-, or a mixture thereof. Embodiment 5. A compound according to any one of embodiments 1 to 4, which is capable of binding to fibroblast activation protein (FAP). Embodiment 6. The compound of any one of embodiments 1 to 5, comprising a diagnostically active nuclide or a therapeutically active nuclide. Embodiment 7. The following formula:

[0041] [ka]

[0042] The compound Hex-[Cys(tMeBn(InDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3590), The following formula

[0043] [ka]

[0044] The compound Hex-[Cys(tMeBn(LuDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3591), The following formula

[0045] [ka]

[0046] The compound Hex-[Cys(tMeBn(GaDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3592), The following formula

[0047] [ka]

[0048] The compound Hex-[Cys(tMeBn(EuDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3661), The following formula

[0049] [ka]

[0050] The compound Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3623), The following formula

[0051] [ka]

[0052] The compound Hex-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3624), The following formula

[0053] [ka]

[0054] The compound Hex-[Cys(tMeBn(EuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3662), The following formula

[0055] [ka]

[0056] The compound Hex-[Cys(tMeBn(GaDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3949), The following formula

[0057] [ka]

[0058] the compound Hex-[Cys-(tMeBn(CuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4293), and The following formula

[0059] [ka]

[0060] The compound Hex-[Cys-(tMeBn(ZnDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4343) The compound of embodiment 6, selected from the group comprising: Embodiment 8. The compound of any one of embodiments 6 and 7, wherein the diagnostically active nuclide is a diagnostically active radionuclide. Embodiment 9. The diagnostically active radionuclide is 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 111 In, 152 Tb, 155Tb, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, preferably 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, most preferably 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124 The compound of embodiment 8, selected from the group consisting of I. Embodiment 10. The compound of any one of embodiments 6 and 7, wherein the therapeutically active nuclide is a therapeutically active radionuclide. Embodiment 11. The therapeutically active radionuclide is 47 Sc, 67 Cu, 89 Sr, 90 Y, 153 Sm, 149 Tb, 161 Tb, 177 Lu, 186 Re, 188 Re, 212 Pb, 213 Bi, 223 Ra, 225 Ac, 226 Th, 227 Th, 131 I, 211 At, preferably 47Sc, 67 Cu, 90 Y, 177 Lu, 188 Re, 212 Pb, 213 Bi, 225 Ac, 227 Th, 131 I, 211 At, most preferably 90 Y, 177 Lu, 225 Ac, 227 Th, 131 I, and 211 The compound of embodiment 10, selected from the group consisting of At. Embodiment 12. The compound of any one of embodiments 1 to 11, which interacts with a fibroblast activation protein (FAP), preferably a human FAP having the amino acid sequence of SEQ ID NO: 1, or a homolog thereof, wherein the amino acid sequence of the homolog has at least 85% identity with the amino acid sequence of SEQ ID NO: 1. Embodiment 13 The compound of embodiment 12, which is an inhibitor of fibroblast activation protein (FAP). Embodiment 14. A compound of any one of embodiments 1 to 13 for use in a method for the diagnosis of a disease. Embodiment 15. The compound for use according to embodiment 14, wherein the disease is a fibroblast activation protein (FAP), preferably a disease involving upregulated expression of fibroblast activation protein (FAP). Embodiment 16. The compound for use according to any one of embodiments 14 to 15, wherein the disease involves cells that exhibit upregulated expression of fibroblast activation protein (FAP), preferably diseased tissues that comprise cells that exhibit upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving fibroblasts associated with a tumor. Embodiment 17. The compound for use according to any one of embodiments 14 to 16, wherein the disease is a neoplasm, preferably a cancer or tumor. Embodiment 18. The compound for use according to embodiment 17, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising solid tumors, epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, and thyroid cancer. Embodiment 19. The compound for use according to embodiment 18, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising breast cancer, colorectal cancer, cholangiocarcinoma, head and neck cancer, lung cancer, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, and squamous cell carcinoma. Embodiment 20. The compound for use according to any one of embodiments 14 to 16, wherein the disease is selected from the group comprising inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases. Embodiment 21 The compound for use according to embodiment 20, wherein the disease is an inflammatory disease. Embodiment 22. The compound for use according to embodiment 21, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis. Embodiment 23 The compound for use according to embodiment 20, wherein the disease is a cardiovascular disease. Embodiment 24 The compound for use according to embodiment 23, wherein the disease is a cardiovascular disease associated with atherosclerotic plaques. Embodiment 25. A compound for use according to embodiment 24, wherein the disease is an atherosclerotic lesion caused by plaque rupture, acute coronary syndrome, myocardial infarction, thrombosis, or vascular occlusion. Embodiment 26 The compound for use according to embodiment 20, wherein the disease is a fibrotic disease. Embodiment 27. The compound for use according to embodiment 26, wherein the disease is selected from the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and hepatic fibrosis. Embodiment 28. A compound for use according to any one of embodiments 14 to 27, comprising a diagnostically active nuclide, preferably a diagnostically active radionuclide. Embodiment 29. The diagnostically active nuclide 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 111 In, 152 Tb, 155 Tb, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, preferably 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, more preferably 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124 The compound for use according to embodiment 28, selected from the group comprising I. Embodiment 30 The compound for use according to any one of embodiments 14 to 29, wherein the method for diagnosis is an imaging method. Embodiment 31 The compound for use according to embodiment 30, wherein the imaging method is selected from the group consisting of scintigraphy, single photon emission computed tomography (SPECT), and positron emission tomography (PET). Embodiment 32. The compound for use according to any one of embodiments 14 to 31, wherein the method comprises administering a diagnostically effective amount of the compound to a subject, preferably a mammal, wherein the mammal is selected from the group comprising humans, companion animals, pets, and livestock, more preferably the subject is selected from the group comprising humans, dogs, cats, horses, and cattle, and most preferably the subject is a human. Embodiment 33 A compound of any one of embodiments 1 to 13 for use in a method for the treatment of a disease. Embodiment 34. The compound for use according to embodiment 34, wherein the disease is a fibroblast activation protein (FAP), preferably a disease in which upregulated expression of fibroblast activation protein (FAP) is involved. Embodiment 35. The compound for use according to any one of embodiments 33 to 34, wherein the disease involves cells exhibiting upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue comprising cells exhibiting upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving fibroblasts associated with a tumor. Embodiment 36. A compound for use according to any one of embodiments 33 to 35, wherein the disease is a neoplasm, preferably a cancer or tumor. Embodiment 37. The compound for use according to embodiment 36, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising solid tumors, epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, and thyroid cancer. Embodiment 38. The compound for use according to embodiment 37, wherein the neoplasms, cancers, and tumors are each individually selected from the group consisting of breast cancer, colorectal cancer, cholangiocarcinoma, head and neck cancer, lung cancer, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, and squamous cell carcinoma. Embodiment 39. The compound for use according to any one of embodiments 33 to 35, wherein the disease is selected from the group comprising inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases. Embodiment 40 The compound for use according to embodiment 39, wherein the disease is an inflammatory disease. Embodiment 41. A compound for use according to embodiment 40, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis. Embodiment 42 The compound for use according to embodiment 39, wherein the disease is a cardiovascular disease. Embodiment 43. The method of claim 43, wherein the disease is a cardiovascular disease associated with atherosclerotic plaques. 43. A compound for use according to embodiment 42. Embodiment 44. A compound for use according to embodiment 43, wherein the disease is an atherosclerotic lesion caused by plaque rupture, acute coronary syndrome, myocardial infarction, thrombosis, or vascular occlusion. Embodiment 45 The compound for use according to embodiment 39, wherein the disease is a fibrotic disease. Embodiment 46. The compound for use according to embodiment 45, wherein the disease is selected from the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and hepatic fibrosis. Embodiment 47. A compound for use according to any one of embodiments 33 to 38, comprising a therapeutically active nuclide, preferably a therapeutically active radionuclide. Embodiment 48. The therapeutically active nuclide is 47 Sc, 67 Cu, 89 Sr, 90 Y, 153 Sm, 149 Tb, 161 Tb, 177 Lu, 186 Re,188 Re, 212 Pb, 213 Bi, 223 Ra, 225 Ac, 226 Th, 227 Th, 131 I, 211 At, preferably 47 Sc, 67 Cu, 90 Y, 177 Lu, 188 Re, 212 Pb, 213 Bi, 225 Ac, 227 Th, 131 I, 211 At, most preferably 90 Y, 177 Lu, 225 Ac, 227 Th, 131 I, and 211 The compound for use according to embodiment 47, selected from the group comprising At. Embodiment 49. The compound for use according to any one of embodiments 33 to 48, wherein the method comprises administering a therapeutically effective amount of the compound to a subject, preferably a mammal, wherein the mammal is selected from the group comprising humans, companion animals, pets, and livestock, more preferably the subject is selected from the group comprising humans, dogs, cats, horses, and cattle, and most preferably the subject is a human. Embodiment 50. A compound according to any one of embodiments 1 to 13 for use in a method for identifying a subject, wherein the subject is likely to respond or not to respond to treatment of a disease, and wherein the method for identifying a subject comprises carrying out a method of diagnosis using a compound according to any one of embodiments 1 to 13, preferably a method for diagnosing a disease as described in any one of embodiments 14 to 33. Embodiment 51. A compound according to any one of embodiments 1 to 13 for use in a method for selecting a subject from a group of subjects, said subject likely to respond or not likely to respond to treatment of a disease, said method for selecting a subject from a group of subjects comprising carrying out a method of diagnosis using a compound according to any one of embodiments 1 to 13, preferably a method for diagnosis of a disease as described in any one of embodiments 14 to 32. Embodiment 52. A compound according to any one of embodiments 1 to 13 for use in a method for stratifying a group of subjects into those likely to respond to treatment of a disease and those unlikely to respond to treatment of a disease, wherein said method for stratifying a group of subjects comprises carrying out a method of diagnosis using a compound according to any one of embodiments 1 to 13, preferably a method for diagnosis of a disease as described in any one of embodiments 14 to 32. Embodiment 53. The compound for use according to any one of embodiments 50 to 52, wherein the disease is a disease involving upregulated expression of fibroblast activation protein (FAP), preferably fibroblast activation protein (FAP). Embodiment 54. The compound for use according to any one of embodiments 50 to 53, wherein the disease involves diseased tissues comprising cells exhibiting upregulated expression of fibroblast activation protein (FAP), preferably cells exhibiting upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving fibroblasts associated with a tumor. Embodiment 55. A compound for use according to any one of embodiments 50 to 54, wherein the disease is a neoplasm, preferably a cancer or tumor. Embodiment 56. The neoplasms, cancers, and tumors are solid tumors, epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, and gastrointestinal stromal tumors. 56. The compound for use according to embodiment 55, wherein the compound is each individually selected from the group comprising: head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, and thyroid cancer. Embodiment 57. The compound for use according to embodiment 56, wherein the neoplasms, cancers, and tumors are each individually selected from the group consisting of breast cancer, colorectal cancer, cholangiocarcinoma, head and neck cancer, lung cancer, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, and squamous cell carcinoma. Embodiment 58. The compound for use according to any one of embodiments 50 to 54, wherein the disease is selected from the group comprising inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases. Embodiment 59. The compound for use according to embodiment 58, wherein the disease is an inflammatory disease. Embodiment 60 The compound for use according to embodiment 59, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis. Embodiment 61 The compound for use according to embodiment 58, wherein the disease is a cardiovascular disease. Embodiment 62. A compound for use according to embodiment 61, wherein the disease is a cardiovascular disease associated with atherosclerotic plaques. Embodiment 63. A compound for use according to embodiment 62, wherein the disease is an atherosclerotic lesion caused by plaque rupture, acute coronary syndrome, myocardial infarction, thrombosis, or vascular occlusion. Embodiment 64 The compound for use according to embodiment 58, wherein the disease is a fibrotic disease. Embodiment 65. The compound for use according to embodiment 64, wherein the disease is selected from the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and hepatic fibrosis. Embodiment 66 The compound for use according to any one of embodiments 50 to 65, wherein the method of diagnosis is an imaging method. Embodiment 67. The compound for use according to embodiment 66, wherein the imaging method is selected from the group comprising scintigraphy, single photon emission computed tomography (SPECT), and positron emission tomography (PET). Embodiment 68. A compound for use according to any one of embodiments 50 to 67, comprising a diagnostically active nuclide, preferably a diagnostically active radionuclide. Embodiment 69. The diagnostically active nuclide 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 111 In, 152 Tb, 155 Tb, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, preferably 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, most preferably 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124The compound for use according to embodiment 68, selected from the group comprising I. Embodiment 70. A compound according to any one of embodiments 1 to 13, for use in a method for delivering an effector to a fibroblast activation protein (FAP), preferably a human fibroblast activation protein (FAP), wherein the effector is selected from the group comprising diagnostically active agents and therapeutically active agents. Embodiment 71. The compound for use according to embodiment 70, wherein the effector is selected from the group comprising diagnostically active nuclides and therapeutically active nuclides. Embodiment 72 The compound for use according to embodiment 71, wherein the diagnostically active nuclide is a diagnostically active radionuclide. Embodiment 73. The diagnostically active radionuclide is 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 1 11 In, 152 Tb, 155 Tb, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, preferably 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76 Br, 77Br, 123 I, 124 I, 125 I, most preferably 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124 The compound for use according to embodiment 72, selected from the group consisting of I. Embodiment 74. The compound for use according to any one of embodiments 70 to 73, wherein said fibroblast activation protein (FAP) is expressed by a cell, preferably a fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell, more preferably a human fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell, most preferably a human fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell, each of which exhibits upregulated expression of fibroblast activation protein (FAP). Embodiment 75. The compound for use according to embodiment 74, wherein the cell is contained in or is part of a tissue, preferably a diseased tissue of a subject suffering from a disease. Embodiment 76. A compound for use according to embodiment 75, wherein the disease involves cells that exhibit upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue comprising cells that exhibit upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving fibroblasts associated with a tumor. Embodiment 77. A compound for use according to any one of embodiments 75 to 76, wherein the disease is a neoplasm, preferably a cancer or tumor. Embodiment 78. The compound for use according to embodiment 77, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising solid tumors, epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, and thyroid cancer. Embodiment 79. The compound for use according to embodiment 78, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising breast cancer, colorectal cancer, cholangiocarcinoma, head and neck cancer, lung cancer, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, sarcoma, and squamous cell carcinoma. Embodiment 80. The compound for use according to any one of embodiments 75 to 76, wherein the disease is selected from the group comprising inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases. Embodiment 81 The compound for use according to embodiment 80, wherein the disease is an inflammatory disease. Embodiment 82. A compound for use according to embodiment 81, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis. Embodiment 83 The compound for use according to embodiment 80, wherein the disease is a cardiovascular disease. Embodiment 84. A compound for use according to embodiment 83, wherein the disease is a cardiovascular disease associated with atherosclerotic plaques. Embodiment 85. A compound for use according to embodiment 84, wherein the disease is an atherosclerotic lesion caused by plaque rupture, acute coronary syndrome, myocardial infarction, thrombosis, or vascular occlusion. Embodiment 86 The compound for use according to embodiment 80, wherein the disease is a fibrotic disease. Embodiment 87. The compound for use according to embodiment 86, wherein the disease is selected from the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and hepatic fibrosis. Embodiment 88 The compound for use according to embodiment 71, wherein the therapeutically active nuclide is a therapeutically active radionuclide. Embodiment 89. The therapeutically active radionuclide is 47 Sc, 67 Cu, 89 Sr, 90 Y, 153 Sm, 149 Tb, 161 Tb, 177 Lu, 186 Re, 188 Re, 212 Pb, 213 Bi, 223 Ra, 225 Ac, 226 Th, 227 Th, 131 I, 211 At, preferably 47 Sc, 67 Cu, 90 Y, 177 Lu, 188 Re, 212 Pb, 213 Bi, 225 Ac, 227 Th, 131 I, 211 At, most preferably 90 Y, 177 Lu, 225 Ac, 227 Th, 131 I, and 211 The compound for use according to embodiment 88, selected from the group consisting of At. Embodiment 90. The compound for use according to any one of embodiments 88 to 89, wherein the fibroblast activation protein (FAP) is expressed by a cell, preferably a fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell, more preferably a human fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell, most preferably a human fibroblast, mesenchymal stem cell, smooth muscle cell, cell of epithelial origin, or endothelial cell that exhibits upregulated expression of fibroblast activation protein (FAP). Embodiment 91. The compound for use according to embodiment 90, wherein the cell is contained in or is part of a tissue, preferably a diseased tissue of a subject suffering from a disease. Embodiment 92. The compound for use according to embodiment 91, wherein the disease involves cells that exhibit upregulated expression of fibroblast activation protein (FAP), preferably diseased tissues that comprise cells that exhibit upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving fibroblasts associated with a tumor. Embodiment 93. A compound for use according to any one of embodiments 90 to 92, wherein the disease is a neoplasm, preferably a cancer or tumor. Embodiment 94. The compound for use according to embodiment 93, wherein the neoplasms, cancers, and tumors are each individually selected from the group comprising solid tumors, epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, cholangiocarcinoma, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and carcinomas, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, and thyroid cancer. Embodiment 95. A composition, preferably a pharmaceutical composition, comprising a compound of any one of embodiments 1 to 13 and a pharmaceutically acceptable excipient. Embodiment 96. The composition of embodiment 95 for use in any method defined in any of the preceding claims. Embodiment 97. A method for the diagnosis of a disease in a subject, comprising administering to the subject a diagnostically effective amount of a compound of any one of embodiments 1 to 13. Embodiment 98 The method of embodiment 97, wherein the compound comprises a diagnostically active agent, the agent preferably being a radionuclide. Embodiment 99. A method for treating a disease in a subject, comprising administering to the subject a therapeutically effective amount of a compound of any one of embodiments 1 to 13. Embodiment 100 The method of embodiment 99, wherein the compound comprises a therapeutically active agent, the agent preferably being a radionuclide. Embodiment 101. The method of any one of embodiments 97 to 100, wherein the disease is a disease involving upregulated expression of fibroblast activation protein (FAP), preferably fibroblast activation protein (FAP). Embodiment 102. The method of any one of embodiments 97 to 101, wherein the disease involves diseased tissue comprising cells that exhibit upregulated expression of fibroblast activation protein (FAP), preferably cells that exhibit upregulated expression of fibroblast activation protein (FAP), more preferably a disease involving tumor-associated fibroblasts. Embodiment 103 The method of any one of embodiments 97 to 102, wherein the disease is selected from the group comprising neoplasia, preferably cancer or tumor, as well as inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases. Embodiment 104. A kit comprising a compound according to any one of embodiments 1 to 13, optionally one or more excipients, and optionally one or more devices. wherein said device is selected from the group comprising a labeling device, a purification device, a handling device, a radiation protection device, an analytical device, or an administration device. Embodiment 105. A kit according to embodiment 104 for use in any method defined in any of the preceding claims.

[0061] More specifically, the problem underlying the present invention is that in a first aspect: The following formula

[0062] [ka]

[0063] the compound Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554), and The following formula

[0064] [ka]

[0065] The compound Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407) The problem is solved by a compound selected from the group consisting of:

[0066] More specifically, in a second aspect, the problem underlying the present invention is solved by a compound according to the first aspect, including any embodiment, for use in a method for the diagnosis of a disease. .

[0067] More particularly, in a third aspect the problem underlying the present invention is solved by a compound according to the first aspect, including any embodiment, for use in a method for the treatment of a disease.

[0068] More particularly, in a fourth aspect the problem underlying the present invention is solved by a compound according to the first aspect including any embodiment for use in a method for identifying a subject, wherein the subject is likely to respond or not to respond to treatment of a disease, the method for identifying a subject comprising performing a method of diagnosis using a compound according to the first aspect including any embodiment.

[0069] More particularly, in a fifth aspect the problem underlying the present invention is solved by a compound according to the first aspect including any embodiment for use in a method of selecting subjects from a population of subjects, wherein the subjects are likely to respond or not to respond to treatment of a disease, the method of selecting subjects from a population of subjects comprising performing a method of diagnosis using a compound according to the first aspect including any embodiment.

[0070] More particularly, in a sixth aspect the problem underlying the present invention is solved by a compound according to the first aspect including any embodiment for use in a method of stratifying a population of subjects into subjects likely to respond to treatment of a disease and subjects unlikely to respond to treatment of a disease, wherein the method for stratifying subjects comprises performing a method of diagnosis using a compound according to the first aspect including any embodiment.

[0071] More specifically, in a seventh aspect, the problem underlying the present invention is solved by a composition, preferably a pharmaceutical composition, comprising a compound according to the first aspect, including any embodiment, and a pharmaceutically acceptable excipient.

[0072] More particularly, in an eighth aspect the problem underlying the present invention is solved by a method for diagnosing a disease in a subject, the method comprising administering to the subject a diagnostically effective amount of a compound according to the first aspect, including any embodiment.

[0073] More particularly, in a ninth aspect the problem underlying the present invention is solved by a method for the treatment of a disease in a subject, the method comprising the step of administering to the subject a therapeutically effective amount of a compound according to the first aspect, including any embodiment.

[0074] More particularly, in a tenth aspect the problem underlying the present invention is solved by a kit comprising a compound according to the first aspect including any embodiment, one or more optional excipient(s), and optionally one or more device(s), wherein the device(s) are selected from the group comprising a labelling device, a purification device, a manipulation device, a radioprotection device, an analytical device or an administration device.

[0075] One of ordinary skill in the art will recognize that a compound or compounds of the present invention is any compound disclosed herein, including, but not limited to, any compound described in any of the above embodiments and any of the following embodiments.

[0076] One of ordinary skill in the art will recognize that a method or methods of the present invention is any method disclosed herein, including, but not limited to, any method described in any of the above embodiments and any of the following embodiments.

[0077] Those of ordinary skill in the art will recognize that a composition or compositions of the present invention is any composition disclosed herein, including, but not limited to, any composition described in any of the above embodiments and any of the following embodiments.

[0078] Those skilled in the art will recognize that a kit or kits of the present invention is any kit disclosed herein, including, but not limited to, any kit described in any of the above embodiments and any of the following embodiments.

[0079] Because no cyclic peptide-based inhibitors specific for fibroblast activation proteins (FAPs) with nanomolar affinity have been previously described, the present invention is based on the inventors' surprising discovery that the compounds of the present invention, and more particularly the cyclic peptides thereof, provide highly specific binding of compounds comprising such cyclic peptides to FAPs.

[0080] Finally, the present inventors have found that the compounds of the present invention are surprisingly stable in plasma, surprisingly useful as imaging agents, and effective in shrinking tumors. In certain embodiments, as preferably used herein, a chelating agent is a compound capable of forming a chelate, whereby the chelating agent is a compound, preferably a cyclic compound, in which a metal or moiety having an electron gap or lone pair participates in the formation of a ring. More preferably, the chelating agent is a compound of this type in which a single ligand occupies more than one coordination site at the central atom.

[0081] In certain embodiments, as preferably used herein, a diagnostically active compound is a compound that is suitable or useful in the diagnosis of disease. In certain embodiments, as preferably used herein, a diagnostic agent or diagnostically active agent is a compound that is suitable or useful in the diagnosis of disease.

[0082] In certain embodiments, as preferably used herein, a therapeutically active compound is a compound that is suitable or useful in the treatment of a disease. In certain embodiments, a therapeutic agent or therapeutically active agent, as preferably used herein, is a compound suitable or useful in the treatment of a disease.

[0083] In certain embodiments, as preferably used herein, a diagnostically therapeutically active compound is a compound that is suitable or useful for both the diagnosis and treatment of a disease. In certain embodiments, as preferably used herein, a diagnostic therapeutic agent or diagnostic therapeutically active agent is a compound that is suitable or useful for both the diagnosis and treatment of a disease.

[0084] In certain embodiments, diagnostic therapeutics, as preferably used herein, is a method for the combined diagnosis and treatment of a disease, preferably wherein the combined diagnostically and therapeutically active compounds used in the diagnostic therapeutics are radiolabeled.

[0085] In certain embodiments, treatment of a disease, as used herein, is treatment and / or prevention of a disease. In an embodiment, as preferably used herein, a disease involving a FAP is a disease in which cells, including but not limited to fibroblasts, that express a FAP, preferably in an upregulated manner, and tissues containing or including cells, such as fibroblasts, that express a FAP, or that express a FAP, respectively, preferably in an upregulated manner, are one or the only cause of the disease and / or symptoms of the disease, or are part of the underlying pathology of the disease. A preferred FAP-expressing cell is a cancer-associated fibroblast (CAF). In disease embodiments, preferably when used in connection with the treatment, treating, and / or therapy of a disease, the effects on cells, tissues, and pathology, respectively, are associated with the onset and progression of the disease. and / or results in a cure, treatment, or amelioration of symptoms of the disease. In disease embodiments, preferably when used in connection with diagnosing and / or diagnosing the disease, the labeling of FAP-expressing cells and / or FAP-expressing tissues allows for the identification or differentiation of said cells and / or tissues from healthy or non-FAP-expressing cells and / or healthy or non-FAP-expressing tissues, respectively. More preferably, such identification or differentiation forms the basis of said diagnosis and diagnosing. In such embodiments, labeling refers to the interaction of a detectable label, directly or indirectly, with FAP-expressing cells and / or FAP-expressing tissues or tissues containing such FAP-expressing cells, more preferably, such interaction involves or is based on the interaction of the label or a compound bearing such label with the FAP.

[0086] In one embodiment, as preferably used herein, a target cell is a cell that expresses a FAP and is the sole or sole cause of a disease and / or symptoms of a disease, or is part of the underlying pathology of a disease.

[0087] In one embodiment, and preferably as used herein, a non-target cell is a cell that does not express a FAP and / or is not the sole or sole cause of the disease and / or symptoms of the disease, or is not part of the underlying pathology of the disease.

[0088] In certain embodiments, as preferably used herein, a neoplasm is an abnormal new growth of cells. Cells in a neoplasm proliferate more rapidly than normal cells and continue to proliferate if not treated. Neoplasms can be benign or malignant.

[0089] In certain embodiments, a tumor, as preferably used herein, is a mass lesion that can be benign or malignant. In certain embodiments, cancer, as preferably used herein, is a malignant neoplasm.

[0090] The amino acid sequence of peptide provided herein is written in typical peptide sequence format, as understood by those skilled in the art.For example, the three-letter designation of conventional amino acid, or the designation of non-conventional amino acid, or the abbreviation of additional component indicates that amino acid or component exists at a specific position in peptide sequence.Each amino acid designation or component is connected to the designation or component of the next and / or previous amino acid in sequence by a hyphen (typically representing amide linkage).

[0091] Where an amino acid contains more than one amino acid and / or carboxy group, all orientations of this amino acid are in principle possible, although in α-amino acids, utilization of the α-amino and α-carboxy groups is preferred; otherwise, the preferred orientation is explicitly specified.

[0092] For amino acids, in their abbreviations, the first letter indicates the stereochemistry of the C-α atom, if applicable, e.g., a capitalized first letter indicates that the L-form of the amino acid is present in the peptide sequence, while a lowercase first letter indicates that the D-form of the corresponding amino acid is present in the peptide sequence.

[0093] In one embodiment, as preferably used herein, an aromatic L-α-amino acid is any type of L-α-amino acid that contains an aryl group. In one embodiment, as preferably used herein, a heteroaromatic L-α-amino acid is any type of L-α-amino acid that contains a heteroaryl group.

[0094] Unless otherwise indicated, amino acid sequences are presented herein in an N-terminal to C-terminal direction. The compounds of the invention typically comprise the amino acid sequences provided herein. Conventional amino acids, also referred to as natural amino acids, are identified by their standard three-letter and one-letter abbreviations, as set forth in Table 2.

[0095] [Table 2]

[0096] Unconventional amino acids, also called unnatural amino acids, are any type of non-oligomeric compound that contains an amino group and a carboxyl group and is not a conventional amino acid. Examples of unconventional amino acids and other building blocks used in constructing compounds of the invention are identified according to their abbreviations or names found in Table 3. The structures of some building blocks are shown along with exemplary reagents for introducing the building block into a peptide (e.g., as a carboxylic acid, etc.), or these building blocks are shown as residues fully attached to another structure, such as a peptide or amino acid. Amino acid structures are shown as explicit amino acids, not as residues of amino acids, as they may appear after implementation into a peptide sequence. Some larger chemical moieties consisting of more than one moiety are also shown for clarity.

[0097] [Table 3-1]

[0098] [Table 3-2]

[0099] [Table 3-3]

[0100] [Table 3-4]

[0101] According to the present application, DOTA is 1,4,7,10-tetraazacyclododecane-1,4, Represents 7,10-tetraacetic acid. Furthermore, those skilled in the art will recognize that the presence of a chelator in a compound of the present invention, unless otherwise stated, includes the possibility that the chelator may be complexed to any metal complex partner, i.e., in principle, any metal that can be complexed by a chelator. The explicitly stated chelator of a compound of the present invention or the general term chelator in connection with a compound of the present invention refers to a chelator that is not complexed in this way, or a chelator to which any metal complex partner is bound, where the metal complex partner is any radioactive or non-radioactive metal complex partner. Preferably, the metal chelator complex, i.e., the chelator to which the metal complex partner is bound, is a stable metal chelator complex.

[0102] Non-radioactive metal chelator complexes have several uses, for example, for assessing properties such as stability or activity that are otherwise difficult to determine. One aspect is that cold variants of radioactive versions of metal complex partners (e.g., non-radioactive gallium, lutetium, or indium complexes described in the Examples) can act as surrogates for radioactive compounds. Furthermore, they are valuable tools for identifying metabolites in vitro or in vivo, as well as for assessing the toxicity properties of compounds of the invention. In addition, metal chelator complexes can be used in binding assays that take advantage of the fluorescent properties of some metal complexes with different ligands (e.g., europium salts).

[0103] It will be appreciated by those skilled in the art that the radionuclide to be coupled or attached to the compounds of the present invention will be selected having regard to the particularities of the disease to be treated and / or diagnosed, respectively, and / or the patient population and patient group to be treated and diagnosed, respectively.

[0104] In embodiments of the present invention, radioactive nuclei are also referred to as radionuclides. Radioactive decay is the process by which the nuclei of unstable atoms lose energy by emitting ionizing particles (ionizing radiation). There are various types of radioactive decay. Decay, i.e., energy loss, occurs when an atom with one type of nucleus, called the parent radionuclide, is transformed into an atom with a different nucleus state or into a different nucleus containing a different number of protons and neutrons. Both of these products are called daughter nuclei. In some decays, the parent and daughter are different chemical elements, and therefore, the decay process results in nuclear transmutation (the creation of atoms of new elements). For example, radioactive decay can be alpha decay, beta decay, and gamma decay. Alpha decay occurs when a nucleus emits an alpha particle (helium nucleus). This is the most common process of emitting nucleons, but in rarer types of decay, the nucleus emits protons or specific nuclei of other elements (in a process called cluster decay). Beta decay occurs when a nucleus loses electrons (β - -decay) or positron (β + Radioactive decay occurs when an excited nucleus undergoes a nuclear transformation (gamma decay) and releases a type of neutrino. In contrast, there are radioactive decay processes that do not produce mutations. The energy of an excited nucleus can be emitted as gamma rays in gamma decay, or used to eject orbital electrons by interaction with the excited nucleus in a process called internal conversion, or used to absorb inner atomic electrons from their electron shells, thereby converting nuclear protons into neutrons, which can cause the emission of electron neutrinos in a process called electron capture (EC), or can be emitted without changing the number of protons and neutrons in a process called isomeric transition (IT). Another form of radioactive decay, spontaneous fission (SF), is found only in very heavy chemical elements, resulting in their spontaneous decomposition into smaller nuclei and a small number of isolated nuclear particles.

[0105] In a preferred embodiment of the present invention, radionuclides are used to label the compounds of the present invention. can be done. In an embodiment of the present invention, the radionuclide is suitable for complexation with a chelating agent to provide a radionuclide chelate complex.

[0106] In further embodiments, one or more atoms of the compounds of the invention are of non-natural isotopic composition; preferably, these atoms are radionuclides, more preferably radionuclides of carbon, oxygen, nitrogen, sulfur, phosphorus and the halogens; these radioactive atoms are typically part of amino acids, optionally halogen-containing amino acids, and / or components, and optionally halogenated components of each of the compounds of the invention.

[0107] In a preferred embodiment of the invention, the radionuclide has a half-life that allows for diagnostic and / or therapeutic medical use, specifically a half-life of between 1 minute and 100 days. In a preferred embodiment of the present invention, the radionuclides have decay energies that allow for diagnostic and / or therapeutic medical use. Specifically, for gamma-emitting isotopes, the decay energies are 0.004 to 10 MeV, preferably 0.05 to 4 MeV, for diagnostic use. For positron-emitting isotopes, the decay energies are 0.6 to 13.2 MeV, preferably 1 to 6 MeV, for diagnostic use. For particle-emitting isotopes, the decay energies are 0.039 to 10 MeV, preferably 0.4 to 6.5 MeV, for therapeutic use.

[0108] In a preferred embodiment of the present invention, the radionuclide is industrially produced for medical use, specifically the radionuclide is available in GMP quality. In a preferred embodiment of the present invention, the daughter nuclide(s) after radioactive decay of the radionuclide are compatible with diagnostic and / or therapeutic medical use. Furthermore, the daughter nuclide(s) are stable or further decay in a manner that does not interfere with or even support diagnostic and / or therapeutic medical use. Representative radionuclides that can be used in connection with the present invention are summarized in Table 4.

[0109] Table 4-1

[0110] Table 4-2

[0111] Table 4-3

[0112] Table 4-4

[0113] Table 4-5

[0114] Table 4-6

[0115] Table 4-7

[0116] Table 4-8

[0117] Table 4-9

[0118] Table 4-10

[0119] Table 4-11

[0120] Table 4-12

[0121] Table 4-13

[0122] Table 4-14

[0123] Table 4-15

[0124] Table 4-16

[0125] Table 4-17

[0126] Table 4-18

[0127] Table 4-19

[0128] Table 4-20

[0129] Table 4-21

[0130] Table 4-22

[0131] Table 4-23

[0132] Table 4-24

[0133] Table 4-25

[0134] Table 4-26

[0135] Table 4-27

[0136] Table 4-28

[0137] Table 4-29

[0138] Table 4-30

[0139] Table 4-31

[0140] [Table 4-32]

[0141] [Table 4-33]

[0142] [Table 4-34]

[0143] In an embodiment of the present invention, the radionuclide is used for diagnostic purposes. Preferably, the radioisotope is, but not limited to, 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 111 In, 152 Tb, 155 Tb, 177 Lu, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I. More preferably, the radionuclide is selected from the group comprising: 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76Br, 77 Br, 123 I, 124 I, 125 I. Even more preferably, the radionuclide is selected from the group comprising: 6 4 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124 I. However, it will also be appreciated by those skilled in the art that the use of the above radionuclides is not limited to diagnostic purposes, but also encompasses their use in therapeutic and diagnostic therapeutics when conjugated to the compounds of the present invention.

[0144] In an embodiment of the invention, a radionuclide is used in therapy. Preferably, the radioisotope is: 47 Sc, 67 Cu, 89 Sr, 90 Y, 111 In, 153 Sm, 149 Tb, 161 Tb, 177 Lu, 186 Re, 188 Re, 212 Pb, 213 Bi, 223 Ra, 225 Ac, 226 Th, 227 Th, 131 I, 211 More preferably, the radioisotope is selected from the group including At. 47 Sc, 67 Cu, 90 Y, 177 Lu, 188 Re, 212 Pb, 213 Bi, 225 Ac, 227 Th, 131 I, 211 Even more preferably, the radionuclide is selected from the group comprising: 90 Y, 177 Lu,225 Ac, 227 Th, 131 I and 211 However, it will also be appreciated by those skilled in the art that the use of the above radionuclides is not limited to therapeutic purposes, but also encompasses their use in diagnostics and diagnostic therapeutics when conjugated to the compounds of the present invention.

[0145] In certain embodiments, the compounds of the present invention are present as pharmaceutically acceptable salts. The "pharmaceutically acceptable salts" of the compounds of the present invention are preferably acid or base salts generally considered in the art to be suitable for use in contact with human or animal tissues without undue toxicity or carcinogenicity, and preferably without irritation, allergic reactions, or other problems or complications. Such salts include mineral and organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. The compounds of the present invention can form internal salts that are also pharmaceutically acceptable salts.

[0146] Suitable pharmaceutically acceptable salts include, but are not limited to, hydrochloric acid, phosphoric acid, bromic acid, malic acid, glycolic acid, fumaric acid, sulfuric acid, sulfamic acid, sulfanilic acid, formic acid, toluenesulfonic acid, methanesulfonic acid, benzenesulfonic acid, ethanedisulfonic acid, 2-hydroxyethylsulfonic acid, nitric acid, benzoic acid, 2-acetoxybenzoic acid, citric acid, tartaric acid, lactic acid, stearic acid, salicylic acid, glutamic acid, ascorbic acid, pamoic acid, succinic acid, fumaric acid, maleic acid, propionic acid, hydroxymaleic acid, hydroiodic acid, phenylacetic acid, alkanoic acid, such as acetic acid, HOOC-(CH2) nExamples of pharmaceutically acceptable salts include salts of acids such as -COOH (n is any integer from 0 to 4, i.e., 0, 1, 2, 3, or 4). Similarly, pharmaceutically acceptable cations include, but are not limited to, sodium, potassium, calcium, aluminum, lithium, and ammonium. Those of ordinary skill in the art will recognize additional pharmaceutically acceptable salts for the compounds provided herein. In general, pharmaceutically acceptable acid or base salts can be synthesized from parent compounds containing a basic or acidic moiety by any conventional chemical method. Briefly, such salts can be prepared by reacting the free acid or free base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or in a mixture of the two. Generally, the use of nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is preferred.

[0147] A "pharmaceutically acceptable solvate" of a compound of the present invention is preferably a solvate of a compound of the present invention formed by the association of one or more solvent molecules with one or more molecules of a compound of the present invention. Preferably, the solvent is one generally considered in the art to be suitable for use in contact with human or animal tissues without undue toxicity or carcinogenicity, preferably without irritation, allergic reaction, or other problems or complications. Such solvents include alcohols, ethers, esters, and amines. Contains organic solvent.

[0148] The "hydrate" of the compound of the present invention is formed by the association of one or more water molecules with one or more molecules of the compound of the present invention. Such hydrates include, but are not limited to, hemihydrate, monohydrate, dihydrate, trihydrate and tetrahydrate. Regardless of the hydrate composition, all hydrates are generally considered to be pharmaceutically acceptable.

[0149] The compound of the present invention has high binding affinity to FAP and high inhibitory activity to FAP.Because of this high binding affinity, the compound of the present invention is effective, useful and / or suitable as targeting agent, and when conjugated to another part, as targeting part.Preferably herein, targeting agent is the agent that interacts with the target molecule, which is the FAP in this case.Therefore, with respect to the cells and tissues targeted by the compound of the present invention, any cells and tissues that express the FAP can be targeted or can be targeted.

[0150] In certain embodiments, the compound interacts with a fibroblast activation protein (FAP), preferably a human FAP having the amino acid sequence of SEQ ID NO: 1, or a homolog thereof, wherein the amino acid sequence of the homolog has at least 85% identity to the FAP of SEQ ID NO: 1. In preferred embodiments, the identity is 90%, preferably 95%, 96%, 97%, 98% or 99%.

[0151] The identity between two nucleic acid molecules can be determined as known to those skilled in the art.More specifically, sequence comparison algorithms can be used to calculate the percent sequence homology of test sequence(s) to reference sequence based on designated program parameters.Test sequence is preferably the sequence or protein or polypeptide that is said to be identical to different protein or peptide, or should be tested to see whether it is identical, and if so, how identical it is, and therefore this different protein or polypeptide is also called reference sequence, and is preferably wild-type protein or polypeptide, more preferably human FAP of SEQ ID NO: 1.

[0152] Optimal alignment of sequences for comparison can be achieved, for example, by the local homology algorithm of Smith & Waterman (Smith et al., Advances in Applied Mathematics, 1981, 2:482), by the Needleman & Wunsch (Needleman et al., J Mol Biol, 1970, 48:443), by the search for similarity method of Pearson & Lipman (Pearson et al., Proc Natl Acad Sci USA, 1988, 85:24444), or by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics software package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by visual inspection.

[0153] One example of an algorithm suitable for determining percent sequence identity is the algorithm used in the Basic Local Alignment Search Tool (hereinafter referred to as "BLAST"). See, e.g., Altschul et al., 1990 (Altschul et al., J Mol Biol, 1990, 215:403) and Altschul et al., 1997 (Altschul et al., Nucleic Acids Res, 1997, 25:3389). Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (hereinafter referred to as "NCBI"). The default parameters used in determining sequence identity using software available from NCBI, such as BLASTN (for nucleotide sequences) and BLASTP (for amino acid sequences), are described in McGinnis et al. (McGinnis et al., Nucleic Acids Res, 2004, 32:W20).

[0154] It is within the scope of the present invention that the compounds of the present invention are used or intended for use in a method for treating the diseases disclosed herein. Such a method preferably comprises administering a therapeutically effective amount of a compound of the present invention to a subject in need thereof. Such a method includes, but is not limited to, curative or adjuvant cancer treatment. It is used as a palliative treatment when a cure is not possible and the purpose is to control the disease locally or to alleviate symptoms, or as a therapeutic treatment when the therapy has a survival benefit and may be curative.

[0155] The methods for treating diseases disclosed herein include the treatment of diseases disclosed herein, including tumors and cancers, and can be used as first therapy, or as second, third, fourth, or last therapy.It is also within the scope of the present invention to combine the compounds of the present invention with additional therapeutic approaches.Those skilled in the art will be aware that the exact treatment intent, including curative, adjuvant, neoadjuvant, therapeutic, or palliative treatment intent, depends on the type, location, and stage of the tumor, as well as the general health of the patient.

[0156] In an embodiment of the invention, the disease is selected from the group consisting of neoplasms not otherwise specified, benign neoplasms, neoplasms of unknown benign or malignant nature, malignant neoplasms, metastatic neoplasms, neoplasms of unknown primary or metastatic nature, benign tumor cells, tumor cells of unknown benign or malignant nature, malignant tumor cells, malignant small cell tumors, malignant giant cell tumors, malignant spindle cell tumors, epithelial neoplasms of unspecified nature, benign epithelial tumors, carcinoma in situ of unspecified nature, metastatic carcinoma of unspecified nature, carcinomatosis, benign epithelioma, malignant epithelioma, large cell carcinoma of unspecified nature, undifferentiated carcinoma of unspecified nature, atypical carcinoma of unspecified nature, pleomorphic carcinoma, giant cell and spindle cell carcinoma, giant cell carcinoma, spindle cell carcinoma, pseudosarcomatous carcinoma, pleomorphic cell carcinoma, spheroid cell carcinoma, multiple small tumors, small cell carcinoma of unspecified nature, oat cell carcinoma, small cell carcinoma, spindle cell type, papillary and squamous neoplasm, papilloma of unspecified nature, in Papillary carcinoma in situ, papillary carcinoma not otherwise specified, verrucous papilloma, verrucous carcinoma not otherwise specified, squamous cell papilloma, squamous cell carcinoma, inverted papilloma, papillomatosis not otherwise specified, squamous cell carcinoma in situ not otherwise specified, squamous cell carcinoma not otherwise specified, metastatic squamous cell carcinoma not otherwise specified, squamous cell carcinoma, keratinizing type not otherwise specified, large cell nonkeratinizing squamous cell carcinoma, small cell nonkeratinizing squamous cell carcinoma, spindle cell squamous cell carcinoma, adenoid squamous cell carcinoma, in cases where stromal invasion is suspected Squamous cell carcinoma in situ, Microinvasive squamous cell carcinoma, Queyrat's erythroplasia, Bowen's disease, Lymphoepithelial carcinoma, Basal cell neoplasm, Basal cell tumor, Basal cell carcinoma not otherwise specified, Multicentric basal cell carcinoma, Localized scleroderma basal cell carcinoma, Fibroepithelial basal cell carcinoma, Basosquamous cell carcinoma, Transforming carcinoma, Yadazoline intraepithelioma, Trichoepithelioma, Trichofolliculoma, Trichilemmomas, Calcifying epithelioma, Transitional cell papilloma and carcinoma, Transitional cell papilloma not otherwise specified, Urothelial papilloma, Transitional cell carcinoma in situ, Transitional cell carcinoma not otherwise specified, Schneiderian papilloma, Inverted transitional cell papilloma, Schneiderian carcinoma, Spindle cell transitional cell carcinoma, Basaloid carcinoma, Cloacal carcinoma, Papillary transitional cell carcinoma, Adenoma and adenocarcinoma, Adenoma not otherwise specified, Bronchial adenoma not otherwise specified, InAdenocarcinoma in situ, Adenocarcinoma not otherwise specified, Metastatic adenocarcinoma not otherwise specified, Scirrhous adenocarcinoma, Gastric fibrosis plastica, Superficial spreading adenocarcinoma, Intestinal adenocarcinoma, Diffuse adenocarcinoma, Monomorphic adenoma, Basal cell adenoma, Islet cell adenoma, Islet cell carcinoma, Insulinoma not otherwise specified, Malignant insulinoma, Glucagonoma not otherwise specified, Malignant glucagonoma, Gastrinoma not otherwise specified, Malignant gastrinoma, Mixed islet cell and exocrine adenocarcinoma, Bile duct adenoma, Cholangiocarcinoma, Bile duct cystadenoma, Bile duct cystadenocarcinoma, Hepatocellular adenoma tumor, hepatocellular carcinoma not otherwise specified, benign hepatocellular cholangiomas, mixed hepatocellular carcinoma and cholangiocarcinoma, trabecular adenoma, trabecular adenocarcinoma, embryonal adenoma, eccrine cutaneous cylindroma, adenoid cystic carcinoma, cribriform carcinoma, adenomatous polyp not otherwise specified, adenocarcinoma in adenomatous polyp, tubular adenoma not otherwise specified, tubular adenocarcinoma, adenomatous polyposis coli, adenocarcinoma in adenomatous polyposis coli, multiple adenomatous polyps, solid tumor not otherwise specified, simple carcinoma, carcinoid tumor not otherwise specified, malignant carcinoid tumor, not otherwise specified Argyrophilic carcinoid tumor, malignant argyrophilic carcinoid tumor, non-argyrophilic carcinoid tumor not otherwise specified, malignant non-argyrophilic carcinoid tumor, malignant mucinous carcinoid tumor, composite carcinoid, pulmonary adenomatosis, bronchioloalveolar adenocarcinoma, alveolar adenoma, alveolar adenocarcinoma, papillary adenoma not otherwise specified, papillary adenocarcinoma not otherwise specified, villous adenoma not otherwise specified, adenocarcinoma in villous adenoma, villous adenocarcinoma, tubulovillous adenoma, chromophobe adenoma, chromophobe carcinoma, acidophilic adenoma, acidophilic carcinoma, mixed acidophilic-basophilic adenoma, mixed acidophilic-basophilic carcinoma, acidophilic adenoma, acidophilic adenocarcinoma, basophilic adenoma, basophilic carcinoma, clear cell adenoma, Clear cell adenocarcinoma of unspecified type, adrenal-like tumor, renal cell carcinoma, clear cell adenofibroma, granular cell carcinoma, chief cell adenoma, aqueous clear cell adenoma, aqueous clear cell adenocarcinoma, mixed cell adenoma, mixed cell adenocarcinoma, lipoadenoma, small follicular adenoma, small follicular adenocarcinoma of unspecified type, well-differentiated small follicular adenocarcinoma, trabecular-type small follicular adenocarcinoma, small follicular adenoma, large follicular adenoma, papillary and follicular adenocarcinoma, non-encapsulated sclerosing carcinoma, multiple endocrine adenomas, juxtaglomerular tumor, adrenocortical adenoma of unspecified type, adrenocortical cell carcinoma, compact cell adrenocortical adenoma, severely pigmented atypical adrenocortical adenoma, clear cell adrenocortical adenoma, glomerular cell adrenocortical adenoma, mixed cell adrenocortical adenoma, Endometrioid adenoma unspecified, endometrioid adenoma, borderline malignant tumor, endometrioid carcinoma, endometrioid adenofibroma unspecified, borderline malignant endometrioid adenofibroma, malignant endometrioid adenofibroma, skin adnexal neoplasm, skin adnexal adenoma, skin adnexal carcinoma, hidradenoma, sweat gland tumor unspecified, sweat gland adenocarcinoma, apocrine adenoma, apocrine gland carcinoma, eccrine acrohidradenoma, eccrine spiradenoma, hidradenoma papilliferum, hidradenoma papilliferum, syringoma unspecified, sebaceous adenoma, sebaceous gland adenocarcinoma, cerumen adenoma, cerumen gland adenocarcinoma, mucoepidermoid neoplasm, mucoepidermoid tumor, mucoepidermoid carcinoma, cystic, mucinous, and serous neoplasm, cystadenoma unspecified, cystadenocarcinoma unspecified, serous cystadenoma unspecified, serous cystadenoma borderline malignant tumor, serous cystadenocarcinoma unspecified, papillary cystadenoma unspecified, papillary cystadenoma borderline malignant tumor, papillary cystadenocarcinoma unspecified, papillary serous cystadenoma unspecified, papillary serous cystadenoma borderline malignant tumor, breast cephalic serous cystadenocarcinoma, unspecified serous superficial papilloma, serous superficial papilloma borderline malignant tumor, serous superficial papillary carcinoma, unspecified mucinous cystadenoma, mucinous cystadenoma borderline malignant tumor, unspecified mucinous cystadenocarcinoma, unspecified papillary mucinous cystadenoma, papillary mucinous cystadenoma borderline malignant tumor,Papillary mucinous cystadenocarcinoma, mucinous adenoma, mucinous adenocarcinoma, pseudomyxoma peritonei, mucin-producing adenocarcinoma, signet ring cell carcinoma, metastatic signet ring cell carcinoma, tubular, lobular, and medullary neoplasm, intraductal carcinoma in situ not otherwise specified, invasive ductal carcinoma, comedo carcinoma, comedo carcinoma in situ not otherwise specified, juvenile breast carcinoma, intraductal papilloma, intraductal papillary adenocarcinoma in situ, intracystic papillary adenoma, intracystic carcinoma in situ, intraductal papillomatosis not otherwise specified, subareolar ductal papillomatosis not otherwise specified, medullary carcinoma with amyloid stroma, medullary carcinoma with lymphocytic stroma, in lobular carcinoma in situ, lobular carcinoma not otherwise specified, invasive ductal carcinoma, inflammatory carcinoma, Paget's disease of the breast, Paget's disease and invasive ductal carcinoma, extramammary Paget's disease, acinar cell neoplasm, acinar cell adenoma, acinar cell tumor, acinar cell carcinoma, complex epithelium Neoplasm, adenosquamous carcinoma, adenolymphoma, squamous metaplastic adenocarcinoma, cartilaginous and osseous metaplastic adenocarcinoma, spindle cell metaplastic adenocarcinoma, apocrine metaplastic adenocarcinoma, benign thymoma, malignant thymoma, special gonadal neoplasm, sex cord-stromal tumor, theca cell tumor unspecified, follicle Theca cell carcinoma, luteinoma not otherwise specified, granulosa cell tumor not otherwise specified, malignant granulosa cell tumor, granulosa cell-theca cell tumor, benign male germinoma, male germinoma not otherwise specified, malignant male germinoma, Sertoli-Leydig cell tumor, ovarian male germinoma, tubular male germinoma not otherwise specified, Sertoli cell carcinoma, lipid-accumulating tubular male germinoma, benign Leydig cell tumor, Leydig cell tumor not otherwise specified, malignant Leydig cell tumor, hilar cell tumor, ovarian lipocytoma, adrenal rest tumor, paraneoplastic tumor Glomus and Glomus Tumor, Paraganglioma Not Otherwise Specified, Malignant Paraganglioma, Sympathetic Paraganglioma, Parasympathetic Paraganglioma, Jugular Body Tumor, Aortic Body Tumor, Carotid Bulb Tumor, Extra-Adrenal Paraganglioma Not Otherwise Specified, Malignant Extra-Adrenal Paraganglioma, Pheochromocytoma Not Otherwise Specified, Malignant Pheochromocytoma, Hemangiosarcoma, Glomus Tumor, Glomus Angioma, Nevus and Melanoma, Pigmented Nevus Not Otherwise Specified, Malignant Melanoma Not Otherwise Specified, Nodular Melanoma, Balloon Cell Nevus, Balloon Cell Melanoma, Halo Nevus , fibrous papules of the nose, nerve nevus, giant cell nevus, amelanocytic nevus, amelanotic melanoma, junctional nevus, malignant melanoma in junctional nevus, precancerous melanosis not otherwise specified, malignant melanoma in precancerous melanosis, Hutchinson's melanoma, malignant melanoma in Hutchinson's melanoma, superficial spreading melanoma, intradermal nevus, compound nevus, giant pigmented nevus, malignant melanoma in giant pigmented nevus, epithelioid nevus and spindle cell nevus, epithelioid melanoma, spindle cell melanoma not otherwise specified,Black spindle cells a type, spindle cell melanoma type b, mixed epithelioid and spindle cell melanoma, blue nevus unspecified, malignant blue nevus, cellular blue nevus, soft tissue tumors and sarcomas unspecified, benign soft tissue tumors, sarcoma unspecified, sarcomatosis unspecified, spindle cell sarcoma, giant cell sarcoma, small cell sarcoma, epithelioid cell sarcoma, fibromatous neoplasm, fibroma unspecified, fibrosarcoma unspecified, myxofibroma, fibromyxofibroma, periosteal fibroma, periosteal fibrosarcoma, fascial fibroma, fascial fibrosarcoma, infantile fibrosarcoma, elastoma, aggressive fibromatosis, abdominal fibromatosis, dendroid fibroma, fibrous histiocytoma unspecified, atypical fibrous tissue Histiocytoma, Malignant fibrous histiocytoma, Fibroxanthoma not otherwise specified, Atypical fibroxanthoma, Malignant fibroxanthoma, Dermatofibroma not otherwise specified, Dermatofibroma protuberans, Dermatofibrosarcoma not otherwise specified, Myxomatous neoplasm, Myxoma not otherwise specified, Myxosarcoma, Lipomatous neoplasm, Lipoma not otherwise specified, Liposarcoma not otherwise specified, Fibrolipoma, Well-differentiated liposarcoma, Fibromyxoid liposarcoma, Myxoid liposarcoma, Round cell liposarcoma, Pleomorphic liposarcoma, Mixed liposarcoma, Intramuscular lipoma, Spindle cell lipoma, Angiomyolipoma, Angiomyoliposarcoma, Angiolipoma not otherwise specified, Infiltrating angiolipoma, Myelolipoma, Hibernation adenoma, Lipoblastomatosis, Myo Neoplastic neoplasm, leiomyoma not otherwise specified, intravascular leiomyomatosis, leiomyosarcoma not otherwise specified, epithelioid leiomyoma, epithelioid leiomyosarcoma, cellular leiomyoma, transforming leiomyoma, angiomyoma, angiomyoma, myoma, myoma, rhabdomyoma not otherwise specified, rhabdomyosarcoma not otherwise specified, pleomorphic rhabdomyosarcoma, mixed rhabdomyosarcoma, embryonal rhabdomyosarcoma, adult rhabdomyosarcoma, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, complex mixed and stromal neoplasm, endometrial stromal sarcoma, endolymphatic stromal endometriosis, adenomyoma, pleomorphic adenoma, malignant mixed tumor not otherwise specified, mixed Müllerian tumor, mesodermal mixed tumor, mesodermal nephroma, unspecified renal tumor Blastoma, epithelial nephroblastoma, mesenchymal nephroblastoma, hepatoblastoma, carcinosarcoma not otherwise specified, embryonal carcinosarcoma, myoepithelioma, benign mesenchymoma, mesenchymoma not otherwise specified, malignant mesenchymoma, embryonal sarcoma, fibroepithelial neoplasm, Brenner tumor not otherwise specified, Brenner tumor borderline malignant, malignant Brenner tumor, fibroadenoma not otherwise specified, intraductal fibroadenoma not otherwise specified, periductal fibroadenoma, adenofibroma not otherwise specified, serous adenofibroma, myxoid adenofibroma, cellular intracanalicular fibroadenoma, cystosarcoma phyllodes not otherwise specified, malignant cystosarcoma phyllodes not otherwise specified, juvenile fibroadenoma, synovial neoplasm, benign synovial tumor, synovial sarcoma not otherwise specified, spindle cell synovial sarcomaEpithelioid synovial sarcoma, biphasic synovial sarcoma, clear cell sarcoma of tendons and aponeuroses, mesothelial neoplasm, benign mesothelioma, malignant mesothelioma, benign fibrous mesothelioma, malignant fibrous mesothelioma, benign epithelioid mesothelioma, malignant epithelioid mesothelioma, benign biphasic mesothelioma, malignant biphasic mesothelioma, adenoid tumor of unspecified type, germ cell neoplasm, dysgerminoma, seminoma of unspecified type, undifferentiated seminoma, spermatocytic seminoma, germ cell tumor, embryonal carcinoma of unspecified type, endodermal sinus tumor, polyembryoma, gonadoblastoma, benign teratoma, teratoma of unspecified type, malignant teratoma of unspecified type, teratocarcinoma, undifferentiated malignant teratoma, mesothelioma Interstitial malignant teratoma, dermoid cyst, malignant transformed dermoid cyst, ovarial goiter not otherwise specified, malignant ovarial goiter, goiter carcinoid, trophoblastic neoplasm, hydatidiform mole not otherwise specified, invasive hydatidiform mole, choriocarcinoma, choriocarcinoma with teratoma, malignant trophoblastic teratoma, mesonephroma, benign mesonephroma, mesonephric tumor, malignant mesonephroma, endosalpingioma, vascular tumor, hemangioma not otherwise specified, angiosarcoma, cavernous hemangioma, venous hemangioma, vine hemangioma, Kupffer cell sarcoma, benign hemangioendothelioma, hemangioendothelioma not otherwise specified, malignant hemangioendothelioma, capillary hemangioma, intramuscular hemangioma, Kaposi's sarcoma, angiokeratoma, Verrucous angiokeratoma, benign hemangiopericytoma, hemangiopericytoma not otherwise specified, malignant hemangiopericytoma, angiofibroma not otherwise specified, hemangioblastoma, lymphangioma, lymphangiomas, lymphangiomas not otherwise specified, lymphangiosarcomas, capillary lymphangiomas, cavernous lymphangiomas, cystic lymphangiomas, lymphangioleiomyomatosis, lymphangioleiomyomatosis, angiolymphangioma, osteoma and osteosarcoma, osteoma not otherwise specified, osteosarcoma not otherwise specified, chondroblastic osteosarcoma, fibroblastic osteosarcoma, telangiectatic osteosarcoma, osteosarcoma in Paget's disease of bone, parosteal osteosarcoma, osteoid osteoma not otherwise specified, osteoblastoma, chondroblastic osteosarcoma Neoplasms, Osteochondroma, Osteochondroma not otherwise specified, Chondroma not otherwise specified, Chondroma not otherwise specified, Chondrosarcoma not otherwise specified, Parosteal Chondroma, Parosteal Chondrosarcoma, Chondroblastoma not otherwise specified, Malignant Chondroblastoma, Mesenchymal Chondrosarcoma, Chondromyxoid fibroma, Giant cell tumor, Giant cell tumor of bone not otherwise specified, Malignant giant cell tumor of bone, Giant cell tumor of soft tissue not otherwise specified, Malignant giant cell tumor of soft tissue, Mixed bone tumor, Ewing's sarcoma, Adamantinoma of long bone, Ossifying fibroma, Odontogenic tumor, Benign odontogenic tumor, Odontogenic tumor not otherwise specified, Malignant odontogenic tumor, Odontoma, Cementoma not otherwise specified, tumor, benign cementoblastoma, cementogenic fibroma, giant cementoma, odontoma not otherwise specified, aggregate odontoma, complex odontoma, ameloblastic fibroodontoma, ameloblastic sarcoma, adenoid odontogenic tumor, calcifying odontogenic cyst, ameloblastoma not otherwise specified, malignant ameloblastoma, odontoma, flat odontogenic tumor, odontogenic myxoma, odontogenic fibroma not otherwise specified, ameloblastic fibroma, ameloblastic fibrosarcoma, odontogenic calcifying epithelioma, mixed tumor, craniopharyngioma, pinealoma, pineocytoma, pineoblastoma, melanotic neuroectodermal tumor, chordoma, glioma, malignant glioma, large Cerebral gliomatosis, mixed glioma, subependymal glioma, subependymal giant cell astrocytoma, choroid plexus papilloma unspecified, malignant choroid plexus papilloma, ependymoma unspecified, undifferentiated ependymoma, papillary ependymoma, myxopapillary ependymoma, astrocytoma unspecified, undifferentiated astrocytoma, plasmatic astrocytoma, hypertrophic Cystic astrocytoma, fibrous astrocytoma, pilocytic astrocytoma, cavernoblastoma unspecified, polar cavernoblastoma, astroblastoma, glioblastoma unspecified, giant cell glioblastoma, glioblastoma with sarcomatous elements, primitive polar cavernoblastoma, oligodendroglioma not otherwise specified, undifferentiated oligodendroglioma, oligodendroglioma unspecified Dendroglioblastoma, medulloblastoma not otherwise specified, desmoplastic medulloblastoma, medullomyoblastoma, cerebellar sarcoma not otherwise specified, teratocytoma, pseudoepithelioma neoplasm, ganglioneuroma, ganglioneuromatosis, neuroblastoma not otherwise specified, medulloepithelioma not otherwise specified, teratoid medulloepithelioma, neuroepithelioma not otherwise specified, cavernous neuroblastoma, ganglioglioma, neurocytoma, Pacinian tumor, retinoblastoma not otherwise specified, differentiated retinoblastoma, undifferentiated retinoblastoma, olfactory nerve tumor, sensory neurocytoma, nasal neuroblastoma, olfactory neuroepithelioma, meningioma, meningioma not otherwise specified, meningiomatosis not otherwise specified, malignant meningioma, meningeal meningeal tumor tumor, fibrous meningioma, psammomatous meningioma, angiomatous meningioma, hemangioblastic meningioma, hemangiopericytic meningioma, transitional meningioma, papillary meningioma, meningeal sarcomatosis, nerve sheath tumor, neurofibroma not otherwise specified, neurofibromatosis not otherwise specified, neurofibrosarcoma, melanotic neurofibroma, plexiform neurofibroma, schwannoma not otherwise specified, schwannomatosis, malignant schwannoma, neuroma not otherwise specified, granular cell tumor and alveolar soft part sarcoma, granular cell tumor not otherwise specified, malignant granular cell tumor, alveolar soft part sarcoma, lymphoma not otherwise specified or diffuse, benign lymphoma-like tumor, malignant lymphoma not otherwise specified, non-Hodgkin's malignant lymphoma,Undifferentiated lymphoma, not otherwise specified, stem cell lymphoma, unspecified rotational cell lymphoma, unspecified lymphosarcoma, unspecified lymphosarcoma, lymphoplasmacytic lymphoma, immunoblastic lymphoma, unspecified mixed lymphocytic-histiocytic lymphoma, unspecified, centroblastic-centrocytic diffuse lymphoma, unspecified follicular-centrocytic lymphoma, unspecified well-differentiated lymphocytic lymphoma, unspecified moderately differentiated lymphocytic lymphoma, unspecified split-type follicular-centrocytic lymphoma, unspecified poorly differentiated lymphocytic lymphoma, unspecified pre-liver lymphoma lymphocytic lymphosarcoma, centroblastic malignant lymphoma not otherwise specified, non-cleaved follicular-centered malignant lymphoma not otherwise specified, reticulum cell sarcoma, reticulum cell sarcoma not otherwise specified, pleomorphic reticulum cell sarcoma, nodular reticulum cell sarcoma, Hodgkin's disease, Hodgkin's disease not otherwise specified, lymphocyte-predominant Hodgkin's disease, mixed-cell Hodgkin's disease, lymphocyte-depleted Hodgkin's disease not otherwise specified, lymphocyte-depleted diffuse fibromatosis-type Hodgkin's disease, lymphocyte-depleted reticular Hodgkin's disease, nodular sclerosing Hodgkin's disease not otherwise specified, cellular phase nodular sclerosing Hodgkin's disease, Hodgkin's granuloma, Hodgkin's granuloma, Hodgkin's sarcoma, nodular lymphoma or Follicular nodular lymphoma, not otherwise specified, nodular mixed lymphocytic-histiocytic lymphoma, centroblastic-centrocytic follicular lymphoma, nodular well-differentiated lymphocytic lymphoma, nodular moderately differentiated lymphocytic lymphoma, follicular cleaved follicular-centrocytic lymphoma, nodular poorly differentiated lymphocytic lymphoma, follicular non-cleaved follicular-centrocytic lymphoma, centroblastic lymphoma, mycosis fungoides, Sézary disease, mixed reticuloendothelial neoplasm, microglioma, malignant histiocytosis, histiocytic medullary reticulosis, Letterer-Siwe disease, plasma cell neoplasm , plasma cell myeloma, benign plasma cell neoplasm, plasmacytoma not otherwise specified, malignant plasma cell neoplasm, mast cell tumor, mast cell tumor not otherwise specified, mast cell sarcoma, malignant mastocytosis, Burkitt tumor, Burkitt tumor, leukemia group, leukemia group not otherwise specified, leukemia not otherwise specified, acute leukemia not otherwise specified, subacute leukemia not otherwise specified, chronic leukemia not otherwise specified, nonleukemic leukemia not otherwise specified, combined leukemia group, combined leukemia, lymphocytic leukemia group, lymphocytic leukemia not otherwise specified, acute lymphocytic leukemia, subacute lymphocytic leukemia, chronic lymphocytic leukemia, nonleukemic lymphocytic leukemia, pre-liver, Lymphocyte leukemia, plasma cell leukemia group, plasma cell leukemia, erythroleukemia group, erythroleukemia, acute erythrocyte, chronic erythrocyte, lymphosarcoma cell leukemia group, lymphosarcoma cell leukemia, myeloid leukemia group, myeloid leukemia not otherwise specified, acute myeloid leukemia, subacute myeloid leukemia, chronic myeloid leukemia, non-leukemic myeloid leukemia, neutrophilic leukemia, acute promyelocytic leukemia, basophilic leukemia group, basophilic leukemia, eosinophilic leukemia group, eosinophilic leukemia, monocytic leukemia In some embodiments, the leukemia is selected from the group consisting of: monocytic leukemia, not otherwise specified monocytic leukemia, acute monocytic leukemia, subacute monocytic leukemia, chronic monocytic leukemia, non-leukemic monocytic leukemia, mixed leukemia group, mast cell leukemia, megakaryocytic leukemia, megakaryocytic myelopathy, myeloid sarcoma, hairy cell leukemia, mixed myeloproliferative lymphoproliferative disorder, polycythemia vera, acute panmyelosis, chronic myeloproliferative disorder, myelosclerosis complicated by myeloid metaplasia, idiopathic thrombocythemia, chronic lymphoproliferative disorder.

[0157] In an embodiment of the invention, the disease is selected from the group consisting of pancreatic tumors, pancreatic adenocarcinoma, tumors of the head of the pancreas, body of the pancreas, tail of the pancreas, pancreatic duct, islets of Langerhans, tumors of the neck of the pancreas, prostate tumors, prostate cancer, prostate, neuroendocrine tumors, breast cancer, tumors of the central breast, upper inner quadrant of the breast, lower inner quadrant of the breast, upper outer quadrant of the breast, lower outer quadrant of the breast, axillary process of the breast, tumors of double lesions of the breast, early-onset breast cancer, parathyroid tumors, myeloma, lung cancer, small cell lung cancer, non-small cell lung cancer, tumors of the main bronchus, tumors of the upper lobe of the lung, tumors of the middle lobe of the lung, tumors of the lower lobe of the lung, colorectal cancer, ascending colon, colon Hepatic flexure, transverse colon, splenic flexure, descending colon, sigmoid colon, colonic overlap, small intestinal tumors, liver tumors, hepatocellular adenoma, hepatocellular carcinoma, hepatocellular cholangiocarcinoma, mixed hepatocellular carcinoma and cholangiocarcinoma (ombined), hepatoblastoma, ovarian cancer, sarcoma, osteosarcoma, fibrosarcoma, gastrointestinal stromal tumors, gastrointestinal tract, gastric cancer, thyroid cancer, medullary thyroid carcinoma, thyroid gland, renal cell carcinoma, renal pelvis, bladder tumors, bladder cancer, bladder trigone, bladder dome, bladder lateral wall, bladder posterior wall, ureteral orifice, urachal tumors, overlapping bladder lesions, basal cell carcinoma, basal cell neoplasms, basal cell tumors, basal cell carcinoma, multicentric basal cell carcinoma Basal cell carcinoma, basaloid carcinoma, basal cell adenoma, squamous cell carcinoma, oral squamous cell carcinoma, laryngeal squamous cell carcinoma, cervical cancer, extracervical, cervical overlap, tumors of the uterine cervix, uterine isthmus, uterine tumor, ovarian tumor, cervical esophagus, thoracic esophagus, abdominal esophagus, upper third of the esophagus, middle third of the esophagus, lower third of the esophagus, tumors of overlapping esophageal lesions, endometrial cancer, head and neck cancer, lymphoma, malignant mesothelioma, mesothelial neoplasm, mesothelioma, fibrous mesothelioma, epithelioid mesothelioma, epithelioid mesothelioma, duodenal cancer, neuroendocrine tumor, neuroendocrine tumor of the lung, pancreas neuroendocrine tumors of the stomach, neuroendocrine tumors of the foregut, neuroendocrine tumors of the midgut, neuroendocrine tumors of the hindgut, gastroenteropancreatic neuroendocrine tumors, neuroendocrine carcinoma, neuroendocrine tumors of the breast, neuroendocrine tumors of the ovary, testicular cancer, thymic carcinoma, tumors of the stomach, fundus, body, antrum, pylorus, lesser curvature, greater curvature, overlapping lesions of the stomach, paraganglioma, ganglioneuroma, melanoma, malignant melanoma, nodular melanoma, amelanotic melanoma, superficial spreading melanoma, epithelioid cell melanoma, spindle cell melanoma, mixed epithelioid cell and spindle cell melanoma.

[0158] In further embodiments, the condition is selected from the group consisting of lateral upper lip, lateral lower lip, lateral lip unspecified, upper lip mucosa, lower lip mucosa, lip mucosa unspecified, lip commissure, overlapping lesions of lip, base of tongue unspecified, dorsal surface of tongue unspecified, tongue border, ventral surface of tongue unspecified, anterior two-thirds of tongue unspecified, lingual tonsil, overlapping lesions of tongue, tongue unspecified, upper gingiva, lower gingiva, unspecified gingiva, anterior floor of mouth, lateral floor of mouth, overlapping lesions of floor of mouth, floor of mouth unspecified, hard palate, soft palate unspecified, uvula, overlapping lesions of palate, palate unspecified, buccal mucosa, oral vestibule, retromolar portion, overlapping lesions of other and unspecified parts of oral cavity, oral cavity unspecified, parotid gland, submandibular gland, sublingual gland, overlapping lesions of major salivary glands, major salivary glands unspecified, tonsil fossa, tonsil pillar, tonsil overlap Lesion, tonsil unspecified, fossa, anterior surface of epiglottis, lateral wall of oropharynx, posterior wall of oropharynx, branchial clefts, overlapping lesions of oropharynx, oropharynx unspecified, superior wall of nasopharynx, posterior wall of nasopharynx, lateral wall of nasopharynx, anterior wall of nasopharynx, overlapping lesions of nasopharynx, unspecified nasopharynx, pyriform sinus, postcricoid, hypopharyngeal surface of aryepiglottic folds, posterior wall of hypopharynx, overlapping lesions of hypopharynx, hypopharynx unspecified, unspecified Pharynx, laryngopharynx, Waldeyer's ring, lips, oral and pharyngeal overlap, cervical esophagus, thoracic esophagus, abdominal esophagus, upper third of esophagus, middle third of esophagus, lower third of esophagus, esophageal overlap, esophagus unspecified, cardia unspecified, fundus, body, antrum, pylorus, lesser curvature unspecified, greater curvature unspecified, gastric overlap, unspecified Stomach, duodenum, jejunum, ileum, Meckel's diverticulum, small intestinal duplication, small intestine not otherwise specified, cecum, appendix, ascending colon, hepatic flexure, transverse colon, splenic flexure, descending colon, sigmoid colon, colonic duplication, colon not otherwise specified, rectosigmoid junction, rectum not otherwise specified, anus not otherwise specified, anal canal, cloacal layer, anorectal and anal canal duplication, liver, intrahepatic bile duct, gallbladder, extrahepatic bile duct, ampulla of Vater, biliary duplication, biliary tract not otherwise specified, pancreatic head, pancreatic body, pancreatic tail, pancreatic duct, islets of Langerhans, pancreatic neck, pancreatic duplication, pancreas not otherwise specified, intestine not otherwise specified, digestive system duplication, unspecified Duplication of gastrointestinal tract, nasal cavity, middle ear, maxillary sinus, ethmoid sinus, frontal sinus, sphenoid sinus, accessory sinus, unspecified accessory sinus, glottis, supraglottis, subglottis, laryngeal cartilage, laryngeal duplication, larynx unspecified, trachea, main bronchi, upper lobe of lung, middle lobe of lung, lower lobe of lung, lung duplication, lung unspecified, thymus, heart, anterior mediastinum, posterior mediastinum, mediastinum unspecified, pleura unspecified, cardiac mediastinal and pleural duplication, upper respiratory tract unspecified, respiratory system and intrathoracic organ duplication, airway unspecified, upper limb long bone joints, upper limb short bone joints, lower limb long bone joints, lower limb short bone joints, limb joints and articular cartilage duplication, limb bones unspecified, Skull and facial bones, mandible, vertebral column, ribs, sternum, clavicle, pelvic bones, overlapping lesions of bone joints and articular cartilage, bone unspecified, blood, bone marrow, spleen, reticuloendothelial system unspecified, hematopoietic system unspecified, skin unspecified lips, eyelids unspecified, external ear, skin of face, skin of scalp and neck, skin of trunk, skin of upper limbs, skin of lower limbs, peripheral nerves of head and neck, peripheral nerves of shoulders and arms, peripheral nerves of legs, peripheral nerves of chest, peripheral nerves of abdomen, peripheral nerves of pelvis, peripheral nerves of trunk, overlapping lesions of peripheral nerves and autonomic nervous system, autonomic nervous system unspecified, retroperitoneum, peritoneum, peritoneum unspecified, overlapping lesions of retroperitoneum and peritoneum, head Syndesmotic tissue, connective tissue of arms, connective tissue of legs, connective tissue of breasts, connective tissue of abdomen, connective tissue of pelvis, connective tissue of trunk unspecified, overlapping lesions of subcutaneous connective tissue and other soft tissues, connective tissue unspecified, nipple, central part of breast, upper inner quadrant of breast, lower inner quadrant of breast, upper outer quadrant of breast, lower outer quadrant of breast, axillary process of breast, overlapping lesions of breast, breast unspecified, labia majora, labia minora, clitoris, overlapping lesions of vulva, vulva unspecified, vagina unspecified, cervix, ectocervix, overlapping lesions of cervix, uterine isthmus, endometrium, myometrium, fundus of uterus,Duplicate lesions of uterine corpus, corpus uteri, uterus unspecified, ovaries, fallopian tubes, broad ligament of uterus, round ligament, parametrium, uterine adnexa, Wolffian bodies, Duplicate lesions of female genital tract, female genital tract unspecified, foreskin, glans penis, body of penis, Duplicate lesions of penis, penis unspecified, prostate, cryptorchidism, descended testis, testis unspecified, epididymis, spermatic cord, scrotum unspecified, tunica vaginalis testis, Duplicate lesions of male genital tract, specified Male genitalia unspecified, kidneys unspecified, renal pelvis, ureters, trigone, bladder dome, lateral bladder wall, posterior bladder wall, ureteral orifice, urachus, bladder overlap, bladder unspecified, urethra, accessory urinary glands, urinary overlap, urinary system unspecified, conjunctiva, cornea unspecified, retina, choroid, ciliary body, lacrimal gland, orbit unspecified, eye and adnexal overlap, eye unspecified, meninges, spinal meninges, meninges unspecified, cerebrum, frontal lobe, lateral Temporal lobe, parietal lobe, occipital lobe, ventricle unspecified, cerebellum unspecified, brainstem, overlapping lesions of brain, brain unspecified, spinal cord, cauda equina, olfactory nerve, optic nerve, auditory nerve, cranial nerve unspecified, overlapping lesions of brain and central nervous system, nervous system unspecified, thyroid, adrenal cortex, adrenal medulla, adrenal gland unspecified, parathyroid gland, pituitary gland, craniopharyngeal duct, pineal gland, carotid body, aortic body, overlapping lesions of endocrine glands and related structures, endocrine glands unspecified , unspecified face or neck, unspecified chest, unspecified abdomen, unspecified pelvis, unspecified upper extremities, unspecified lower extremities, other unspecified locations, overlapping lesions in unspecified locations, facial-head-and-neck lymph nodes, intrathoracic lymph nodes, intraperitoneal lymph nodes, axillary-arm lymph nodes, inguinal-leg lymph nodes, pelvic lymph nodes, lymph nodes in multiple regions, unspecified lymph nodes, unknown primary site.

[0159] Subjects treated with the compounds disclosed and claimed herein can be treated in combination with other non-surgical anti-proliferative (e.g., anti-cancer) drug therapies. In one embodiment, the compounds may be administered in combination with an anti-cancer compound, such as a cytostatic compound. A cytostatic compound is a compound (e.g., a small molecule, nucleic acid, or protein) that inhibits cell growth and / or proliferation. In some embodiments, the cytostatic compound is directed against malignant cells of a tumor. In yet other embodiments, the cytostatic compound is directed against vascular smooth muscle cells. It inhibits the growth and / or proliferation of muscle cells or fibroblasts.

[0160] Suitable antiproliferative or cytostatic compounds for use with the compounds disclosed and claimed herein include anti-cancer drugs.Some anti-cancer drugs that can be used are well known and include, but are not limited to, acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; amethanthrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacytidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; Bleomycin sulfate;Brequinar sodium;Bropirimine;Busulfan;Cactinomycin;Calsterone;Caracemide;Carbetimer;Carboplatin;Carmustine;Carubicin hydrochloride;Carzelesin;Cedefingol;Chlorambucil;Ciloremycin;Cisplatin;Cladribine;Crisnatol mesylate;Cyclophosphamide;Cytarabine;Dacarbazine;Dactinomycin;Daunorubicin hydrochloride;Decitabine;Dexormaplatin;Dezaguanine;Dezaguanine mesylate;Diaziquone;Doceta Doxorubicin; Doxorubicin hydrochloride; Droloxifene; Droloxifene citrate; Dromostanolone propionate; Duazomycin; Edatrexate; Eflornithine hydrochloride; Elsamitrucin; Enloplatin; Enpromate; Epipropizine; Epirubicin hydrochloride; Elbrozole; Esorubicin hydrochloride; Estramustine; Estramustine phosphate sodium; Etanidazole; Etoposide; Etoposide phosphate; Etoprine; Fadrozole hydrochloride; Fazarabine; Fenretinide; Flocc Uridine; Fludarabine phosphate; Fluorouracil; Flurocitabine; Fosquidone; Fostriecin sodium; Gemcitabine; Gemcitabine hydrochloride; Hydroxyurea; Idarubicin hydrochloride; Ifosfamide; Irmofosine; Interferon alfa-2a; Interferon alfa-2b; Interferon alfa-n1; Interferon alfa-n3; Interferon beta-1a; Interferon gamma-1b; Iproplatin; Irinotecan hydrochloride; Lanreotide acetate; Letrozole;Leuprolide acetate; Liarozole hydrochloride; Lometrexol sodium; Lomustine; Losoxantrone hydrochloride; Masoprocol; Maytansine; Mechlorethamine hydrochloride; Megestrol acetate; Melengestrol acetate; Melphalan; Menogaril; Mercaptopurine; Methotrexate; Methotrexate sodium; Metoprine; Meturedepa; Mitindomide; Mitocalcin; Mitochromin; Mitogillin; Mitomarcin; Mitomycin; Mitospar; Mitotane; Mitoxantrone hydrochloride; Mycophenolic acid; Niraparib; Nocodazole; Nogalamycin; Olparib; Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Periomycin; Pentamustine; Peplomycin sulfate; Perfosfamide; Pipobroman; Piposulfan; Piroxantrone hydrochloride; Plicamycin; Promestane; Porfimer sodium; Porfiromycin; Prednimustine; Procarbazine hydrochloride; Puromycin; Puromycin hydrochloride; Pirazofurin; Ribopurin; Rogletimide; Rucaparib; Safi Ngol; Safingol hydrochloride; Semustine; Simtrazene; Sparfosate sodium; Sparsomycin; Spirogermanium hydrochloride; Spiromustine; Spiroplatin; Streptonigrin; Streptozocin; Surofenur; Talazoparib; Tallysomycin; Taxol; Taxotere; Tecogalan sodium; Tegafur; Teloxantrone hydrochloride; Temoporfin; Teniposide; Teloxylon; Testolactone; Thiamiprine; Thioguanine; Thiotepa; Tiazofurin; Tirapazamine; Topotecan hydrochloride; toremifene citrate; trestron acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; tuburozole hydrochloride; uracil mustard; uredepa; vapreotide; veraparib; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; binepidine sulfate; vinglisinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrocidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; and zorubicin hydrochloride; Examples include:

[0161] Other anti-cancer drugs include, but are not limited to, 20-epi-1,25-dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; acylfulvene; adecipenol; adozelesin; ALL-TK antagonists; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; anagrelide; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1 (anti-dorsalizing morphogenetic protein-1) protein-1); antiestrogens; antineoplastons; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulaculin; atamestane; atlimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxins; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR / ABL antagonists; benzochlorins; benzoylstaurosporines; beta-lactam derivatives; beta-arretin; betaclamycin B; betulinic acid; bFGF inhibitors; bisaziridinylspermine; bisnafide; bisstraten A; brefullate; budotitanium; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamido-amino-triazoles; carboxyamidotriazoles; CaRest M3; CARN 700; cartilage-derived inhibitor; casein kinase inhibitor (ICOS); castanospermine; cecropin B; cetrorelix; chlorin; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; clomiphene analog; clotrimazole; colismycin A; colismycin B; combretastatin A4; combretastatin analog; conagenin; crambecidin 816; crisnatol; cryptophycin 8; cryptophycin A derivative; curacin A; cyclopentaanthraquinone; cycloplatam; sipemycin; cytarabine ocphosphate; cytolytic factors; cytostatin; dacliximab; dehydrodidemnin B; deslorelin;Dexphosphamide; Dexrazoxane; Dexverapamil; Didemnin B; Didox; Diethylnorspermine; Dihydro-5-azacytidine; 9-dihydrotaxol; Dioxamycin; Diphenylspiromustine; Docosanol; Dolasetron; Doxifluridine; Dronabinol; Duocarmycin SA; Ebselen; Ecomustine; Edelfosine; Edrecolomab; Eflomitine; Elemene; Emiteflu; Epirubicin; Epristeride; Estramustine Narog; Estrogen agonists; Estrogen antagonists; Etanidazole; Etoposide phosphate; Exemestane; Filgrastim; Finasteride; Flavopiridol; Flezelastine; Fluasterone; Fludarabine; Fluorodaunornithine hydrochloride; Forfenimex; Formestane; Fotemustine; Gadolinium texaphyrin; Gallium nitrate; Galocitabine; Ganirelix; Gelatinase inhibitors; Glutathione inhibitors; Hepsulfam; Heregulin; Hexamethylene bisacetamide; Hypericum Synthon; Ibandronic acid; Idoxifene; Idramanton; Ilmofosine; Ilomastat; Imidazoacridone; Imiquimod; Immunostimulatory peptides; Insulin-like growth factor-I receptor inhibitors; Interferon agonists; Interferons; Interleukins; Iobengan; Iododoxorubicin; 4-ipomeanol (ipomeanol, 4-); Irinotecan; Ilopract; Irsogladine; Isobengazole; Isohomohalichondrin B; Itasetron; Jasplakinolide; Kahalalide F; Lamellar Phosphate-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; leukemia inhibitory factor; leukocyte alpha interferon; leuprolide + estrogen + progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogs; lipophilic disaccharide peptides; lipophilic platinum compounds; lysoclinamide 7; lobaplatin; lombricin; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lutotecan; lutetium texaphyrin; ;Lisofylline;Lytic peptides;Maytansine;Mannostatin A;Marimastat;Masoprocol;Maspin;Matrilysin inhibitors;Matrix metalloproteinase inhibitors;Melbarone;Meterelin;Methioninase;Metoclopramide;MIF inhibitors;Mifepristone;Miltefosine;Milimostim;Mismatched double-stranded RNA;Mitoguazone;Mitolactol;Mitomycin analogs;Mitonafide;Mitotoxin fibroblast growth factor-saporin;Mofalotene;Monoclonal antibodies, human chorionic gonadotropin;Monophosphoryl lipids A+ Mycobacterium cell wall SK; Mopidamol; Multidrug resistance gene inhibitors; Multiple tumor suppressor 1-based therapy; Mustard anticancer compounds; Mycaperoxide B; Mycobacterial cell wall extract; Myriaporone; N-acetyldinaline; N-substituted benzamides; Nafarelin; Nagrestip; Naloxone + Pentazocine; Napavine; Nafterpine; Nartograstim; Nedaplatin; Nemorubicin; Neridronic acid; Neutral endopeptidase; Nilutamide; Nisamycin; Nitric oxide modulators; Nitroxide Antioxidants; Nitrulline; O6-benzylguanine; Octreotide; Oxenone; Oligonucleotides; Onapristone; Ondansetron; Ondansetron; Oracin; Oral cytokine inducers; Osateron; Oxaliplatin; Oxaunomycin; Paclitaxel analogs; Paclitaxel derivatives; Palauamine; Palmitoylrhizoxin; Pamidronic acid; Panaxytriol; Panomyphen; Parabactin; Pazeliptin; Pegaspargase; Perdecin; Pentosan polysulfate sodium; Pentostatin; Pentrozole; Perflubron; Perfosfamide; Perillyl alcohol; Phenazinomycin; Phenylacetate; Phosphatase inhibitors; Picibanil; Pilocarpine hydrochloride; Pirarubicin; Piritrexim; Prasetin A; Prasetin B; Plasminogen activator inhibitors; Platinum complexes; Platinum compounds; Platinum-triamine complexes; Porfimer sodium; Porfiromycin; Propylbis-acridone; Prostaglandin J2; Proteasome inhibitors; Protein A-based immunomodulators; Protein kinase C inhibitors; Protein kinase C inhibitor(s); Microalgae;Protein tyrosine phosphatase inhibitors; Purine nucleoside phosphorylase inhibitors; Purpurin; Pyrazoloacridine; Pyridoxylated hemoglobin polyoxyethylene conjugates; RAF antagonists; Raltitrexed; Ramosetron; Ras farnesyl protein transferase inhibitors; Ras inhibitors; Ras-GAP inhibitors; Demethylated reteriptin; Rhenium etidronate (Re186); Rhizoxin; Ribozyme; RII retinamide; Rohitucin; Romurtide; Roquinimex; Rubiginone B1; Ruboxil; Saito Pin; SarCNU; Saclophytol A; Sargramostim; Sdi1 mimetic; Senescence-derived inhibitor 1; Sense oligonucleotide; Signal transduction inhibitor; Signal transduction modulator; Single-chain antigen binding protein; Schizofuran; Sobuzoxane; Sodium borocaptate; Sodium phenylacetate; Sorbrol; Somatomedin-binding protein; Sonermin; Sparfosic acid; Spicamycin D; Spiromustine; Splenopentin; Spongistatin 1; Squalamine; Stem cell inhibitor; Stem cell division inhibitor; Stipiamide; Stromelysin inhibitor; Sulfinosine; superactive vasoactive intestinal peptide antagonist; thradista; suramin; swainsonine; synthetic glycosaminoglycans; talimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellapyrylium; telomerase inhibitors; temozolomide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thalidomide; thiocoraline; thrombopoietin; thrombopoietin mimetics; thymalfasin; thymopoietin receptor agonists; thymotrin; thyroid-stimulating hormone; estrogen These include tylethiopurinase; titanocene dichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; variolin B; vector systems, erythrocyte gene therapy; veraresol; veramine; verudin; vinorelbine; vinxartin; vitaxin; zanoterone; zilascorub; and zinostatin stimalamer.

[0162] The compounds disclosed and claimed herein may also be used in combination with any of the following treatments: Therapy in combination with inhibitors of poly(ADP-ribose) polymerase (PARP), a class of chemotherapy drugs that target cancers with defective DNA damage repair (Yuan et al., Expert Opin Ther Pat, 2017, 27:363). Such PARP inhibitors include, but are not limited to, olaparib, rupacarib, velaparib, niraparib, talazoparib, pamiparib, iniparib, E7449, and A-966492.

[0163] For example, nuclear factor-kappa B signal transduction therapy is combined with inhibitors of signal transduction pathways and mechanisms that lead to the repair of DNA single-strand and double-strand breaks (Pilie et al., Nat Rev Clin Oncol, 2019, 16:81; Zhang et al., Chin J Cancer, 2012, 31:359). Such inhibitors include, but are not limited to, inhibitors of ATM and ATR kinase, checkpoint kinase 1 and 2, DNA-dependent protein kinase, and WEE1 kinase (Pilie et al., Nat Rev Clin Oncol, 2019, 16:81).

[0164] Immunomodulators (Khalil et al., Nat Rev Clin Oncol, 2016, 13:394), cancer vaccines (Hollingsworth et al., NPJ Vaccines, 2019, 4:7), immune checkpoint inhibitors (e.g., PD-1, PD-L1, CTLA-4 inhibitors) (Wei et al., Cancer Discov, 2018, 8:1069), cyclin D kinase 4 / 6 inhibitors (Goel et al., Trends Cell Biol, 2018, 28:911), antibodies that can bind to tumor cells and / or metastases and induce antibody-dependent cellular cytotoxicity (ADCC) (Kellner et al., Transfus Med Hemother, 2017, 44:327), T cell or NK cell engagers (e.g., bispecific antibodies) (Yu et al., J Cancer Res Clin Oncol, 2019, 145:941), in combination with cell therapy using expanded autologous or allogeneic immune cells (e.g., chimeric antigen receptor T (CAR-T) cells) (Khalil et al., Nat Rev Clin Oncol, 2016, 13:394). Immune checkpoint inhibitors include, but are not limited to, nivolumab, ipilimumab, pembrolizumab, atezolizumab, avelumab, durvalumab, and cemiplimab.

[0165] According to the present invention, the compound can be administered before, simultaneously with, or after other anti-cancer compounds. The administration schedule can include administering different agents in an alternating manner. In other embodiments, the compound can be delivered before and during, during and after, or before and after treatment with other therapies. In some cases, the compound is administered more than 24 hours before the administration of other anti-proliferative treatments. In other embodiments, more than one anti-proliferative therapy can be administered to the subject. For example, the subject can receive the compound of the present invention in combination with both surgery and at least one other anti-proliferative compound. Alternatively, the compound can be administered in combination with more than one anti-cancer drug.

[0166] In one embodiment, the compounds of the present invention are used to detect cells and tissues that overexpress FAP, whereby such detection is achieved by conjugating a detectable label, preferably a detectable radionuclide, to the compounds of the present invention. In a preferred embodiment, the detected cells and tissues are diseased cells and tissues and / or are the sole cause of the disease and / or symptoms of the disease, or are part of the underlying pathology of the disease. In a further preferred embodiment, the detected cells and tissues are diseased cells and tissues. The cells and tissues involved cause and / or are part of oncological indications (e.g., neoplasia, tumors, and cancer) or non-oncological indications (e.g., inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases).

[0167] In another embodiment, the compounds of the present invention are used to treat cells and tissues that overexpress FAP. In a preferred embodiment, the treated cells and tissues are diseased cells and tissues, and / or are one or the only cause of disease and / or disease symptoms, or are part of the underlying pathology of disease. In a more preferred embodiment, the diseased cells and tissues cause and / or are part of oncological adaptations (e.g., neoplasia, tumor, and cancer), and therapeutic activity is achieved by conjugating a therapeutically active effector, preferably a therapeutically active radionuclide, to the compounds of the present invention. In a more preferred embodiment, the diseased cells and tissues cause and / or are part of non-oncological adaptations (e.g., inflammatory diseases, cardiovascular diseases, autoimmune diseases, and fibrotic diseases), and therapeutic activity is achieved by inhibiting the enzymatic activity of FAP.

[0168] In further embodiments, particularly when the disease is a non-tumor disease or non-tumor indication (e.g., inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease), the compounds of the present invention are administered in a therapeutically effective amount; preferably, the compounds of the present invention do not contain therapeutically active nuclides. An effective amount is the amount of compound administered that is sufficient to produce a therapeutically or medically desired result or effect in the subject to which the compound is administered. The effective amount will vary depending on the particular condition being treated, the age and physical condition of the subject being treated, the severity of the condition, the duration of treatment, the nature of concurrent or concomitant therapy (if any), the particular route of administration, and factors within the knowledge and expertise of the medical professional. For example, in the context of a method for treating a subject with a condition characterized by abnormal cell proliferation, an effective amount for inhibiting proliferation would be an amount sufficient to reduce or completely stop abnormal cell proliferation, for example, to slow or stop the development or progression of a cell mass such as a tumor. As used in this embodiment, "inhibit" encompasses all of the above.

[0169] In other embodiments, a therapeutically effective amount will be the amount necessary to prolong the dormancy of micrometastases or stabilize remaining primary tumor cells following surgery or drug therapy. Generally, when using unconjugated compounds that do not contain a therapeutically active radionuclide, the therapeutically effective amount will vary depending on the age, condition, and sex of the subject, as well as the nature and extent of the disease in the subject, all of which can be determined by one of ordinary skill in the art. Dosages can be adjusted by the individual physician or veterinarian, particularly in the event of complications. A therapeutically effective amount typically ranges from 0.1 μg / kg to about 2000 mg / kg, or 1.0 μg / kg to about 1000 mg / kg, or about 0.1 mg / kg to about 500 mg / kg, or about 1.0 mg / kg to about 100 mg / kg, administered in one or more doses per day for one or more days. If desired, the effective daily dose of the active compound may be administered as two, three, four, five, six, or more subdoses, for example, administered separately at appropriate intervals throughout the day, in unit dosage forms, as needed. In some embodiments, the compound is administered for more than 7 days, more than 10 days, more than 14 days, and more than 20 days. In still other embodiments, the compound is administered for several weeks or several months. In still other embodiments, the compound is delivered every other day. For example, the agent is delivered every 2 days, or every 3 days, or every 4 days, or every 5 days, or every 6 days, or every week, or every month.

[0170] In a preferred embodiment, the compounds of the invention are for use in the treatment and / or prevention of disease, whereby such treatment is radionuclide therapy. Preferably, radionuclide therapy involves the use of different forms of radiation emitted by radionuclides. Such radiation may be, for example, photon radiation, but is not limited to, β -The radiation may be any one of electron radiation, including particle and Auger electron radiation, proton radiation, neutron radiation, positron radiation, alpha particle radiation or ion beam radiation.Depending on the type of particle or radiation emitted by the radionuclide, radionuclide therapy can be distinguished, for example, as photon-emitting nuclide therapy, electron-emitting nuclide therapy, proton-emitting nuclide therapy, neutron-emitting nuclide therapy, positron-emitting nuclide therapy, alpha particle-emitting nuclide therapy or ion beam radionuclide therapy.All these forms of radionuclide therapy are encompassed by the present invention, and all these forms of radionuclide therapy can be realized by the compound of the present invention, preferably under the condition that the radionuclide attached to the compound of the present invention, more preferably as an effector, provides this type of radiation.

[0171] Radionuclide therapy preferably works by damaging the DNA of cells. Damage is caused by photons, electrons, protons, neutrons, positrons, alpha particles, or ion beams, which directly or indirectly ionize the atoms that make up the DNA chain. Indirect ionization occurs as a result of the ionization of water, forming free radicals, particularly hydroxyl radicals, which then damage DNA.

[0172] In the most common forms of radionuclide therapy, many of the effects of radiation are due to free radicals. Because cells have mechanisms for repairing DNA damage, breaking DNA on both strands proves to be the most significant technique for altering cellular characteristics. Because cancer cells are generally undifferentiated and stem cell-like, they replicate more and have a reduced ability to repair sublethal damage compared to many healthy, differentiated cells. DNA damage is inherited through cell division, causing cancer cells to accumulate damage and die or replicate more slowly.

[0173] Oxygen is a potent radiosensitizer, increasing the effectiveness of a given dose of radiation by forming DNA-damaging free radicals. Therefore, the use of hyperbaric oxygen tanks, blood substitutes that deliver large amounts of oxygen, hypoxic cell radiosensitizers such as misonidazole and metronidazole, and hypoxic cell toxins such as tirapazamine can be applied.

[0174] Other factors to consider when selecting the radioactive dose include whether the patient is receiving chemotherapy, whether the radiation therapy is administered before or after surgery, and the degree of success of the surgery.

[0175] The total radioactive dose can be fractionated, i.e., spread over time as one or more treatments, for several important reasons. Fractionation allows normal cells time to recover, while tumor cells generally repair less efficiently between fractionated doses. Fractionation also allows tumor cells that were in a relatively radioresistant phase of the cell cycle during one treatment to cycle back to a sensitive phase before the next fractionated dose is administered. Similarly, tumor cells that were chronically or acutely hypoxic and therefore more radioresistant can reoxidize between fractions, improving tumor cell killing.

[0176] It is generally known that different cancers respond differently to radiation therapy. The response of a cancer to radiation is described by its radiosensitivity. Cancer cells that are highly radiosensitive are rapidly killed by moderate doses of radiation. These include leukemia, many lymphomas, and germ cell tumors.

[0177] To some extent, it is important to distinguish the radiosensitivity of a particular tumor, which is a laboratory measurement, from the "cure potential" of the cancer by an internally delivered radioactive dose in actual clinical practice. For example, leukemia is generally not curable with radiation therapy because it spreads throughout the body. Lymphoma may be curable with radiation if it is localized to one area of ​​the body. Similarly, many common, moderately radioresponsive tumors can be treated with curative doses of radioactivity when they are in their early stages. This is true, for example, for non-melanoma skin cancer, head and neck cancer, non-small cell lung cancer, cervical cancer, anal cancer, and prostate cancer.

[0178] A tumor's response to radiation therapy is also related to its size. For complex reasons, very large tumors are less responsive to radiation than small tumors or microscopic disease. Various strategies are used to overcome this effect. The most common technique is surgical resection before radiation therapy. This is most commonly seen in the treatment of breast cancer with wide local excision or mastectomy, followed by adjuvant radiation therapy. Another method is to shrink the tumor using neoadjuvant chemotherapy before radionuclide therapy. A third technique is to enhance the cancer's radiosensitivity by administering certain drugs during the course of radiation therapy. Examples of radiosensitizing drugs include, but are not limited to, cisplatin, nimorazole, and cetuximab.

[0179] Intraoperative radiation therapy is a special type of radiation therapy delivered immediately after the surgical removal of the cancer. This method has been used in breast cancer (targeted intraoperative radiation therapy), brain tumors, and rectal cancer.

[0180] Radionuclide therapy itself is painless. Many low-dose palliative treatments are minimally or ineffective. Higher-dose treatments can cause side effects that vary during treatment (acute side effects), within months or years after treatment (long-term side effects), or even after further treatment (cumulative side effects). The nature, severity, and persistence of side effects depend on the organ receiving the radiation, the treatment itself (type of radionuclide, dose, fractionation, concurrent chemotherapy), and the patient.

[0181] It is within the scope of the present invention that each and any of the above strategies may be implemented, insofar as the methods for treating the diseases of the present invention are known as such in the art and constitute further embodiments of the present invention.

[0182] It is also within the scope of the present invention that the compounds of the present invention be used in methods for the diagnosis of the diseases disclosed herein, preferably comprising the step of administering a diagnostically effective amount of a compound of the present invention to a subject in need thereof.

[0183] According to the present invention, the imaging method is selected from the group consisting of scintigraphy, single photon emission computed tomography (SPECT) and positron emission tomography (PET). Scintigraphy is a form of diagnostic examination or method used in nuclear medicine, in which a radiopharmaceutical is internalized by cells, tissues and / or organs, preferably in vivo, and the radiation emitted by the internalized radiopharmaceutical is captured by an external detector (gamma camera) to form and display a two-dimensional image.In contrast, SPECT and PET form and display a three-dimensional image.For this reason, SPECT and PET are classified as separate technologies from scintigraphy, but they also use a gamma camera to detect internal radiation.Scintigraphy is different from diagnostic X-ray, in which external radiation passes through the body to form an image.

[0184] Single Photon Emission Computed Tomography (SPECT) scans are a type of nuclear imaging technique that uses gamma rays. They are very similar to conventional nuclear medicine 2D imaging, which uses a gamma camera. Before a SPECT scan, the patient is injected with a radiolabeled chemical that emits gamma rays that can be detected by the scanner. A computer reads the radiation from the gamma camera. Information is collected and converted into two-dimensional cross-sectional planes. These cross-sectional planes can be reconstructed to form a three-dimensional image of the organ or tissue. SPECT involves the detection of gamma rays emitted by radionuclides provided by radiolabeled chemicals, both singly and sequentially. To acquire a SPECT image, a gamma camera is rotated around the patient. Projection images are acquired at defined points during the rotation, typically every 3–6°. Often, a full 360° rotation is used to obtain optimal reconstruction. The time required to acquire each projection image is also variable, but 15–20 seconds is typical, giving a total scan time of 15–20 minutes. Multi-head gamma cameras are faster. SPECT acquisition is very similar to two-dimensional gamma camera imaging, so the same radiopharmaceuticals can be used.

[0185] Positron emission tomography (PET) is a noninvasive diagnostic imaging technique for measuring the biochemical state or metabolic activity of cells in the human body. PET is unique because it produces images of basic biochemistry or function within the body. Traditional diagnostic techniques, such as X-rays, CT scans, or MRIs, produce images of the body's anatomy or structure. The premise of these techniques is that they can visualize changes in the structure or structure associated with disease. Biochemical processes are also altered by disease and may occur before overall changes in the anatomy. PET is an imaging technique that can visualize some of these early biochemical changes. PET scanners rely on radiation emitted by the patient to create images. Each patient is given a minute amount of a radiopharmaceutical that closely resembles a natural substance used by the body or that specifically binds to a receptor or molecular structure. As a radioisotope undergoes positron-emitting decay (also known as beta-plus decay), it emits a positron, the antiparticle counterpart of the electron. After traveling up to a few millimeters, the positron encounters an electron, annihilates, and generates a pair of counter-moving (gamma) photons. These are detected when they reach the scintillation material in the scanning device, producing a flash of light that is detected by a photomultiplier tube or silicon avalanche photodiode. The technique relies on the simultaneous or coincident detection of photon pairs. Photons that do not arrive as a pair, i.e., within a few nanoseconds of each other, are ignored. All coincidences are forwarded to an image processing unit, where the final image data is produced using an image reconstruction procedure.

[0186] SPECT / CT and PET / CT are the combination of SPECT and PET with computed tomography (CT). A key benefit of combining these modalities is improved reader confidence and accuracy. With traditional PET and SPECT, the limited number of photons emitted from abnormal regions results in very low levels of background that are difficult to anatomically localize to the region. The addition of CT helps determine the location of abnormal regions from an anatomical perspective and classify the likelihood that this represents disease.

[0187] It is within the scope of the present invention that each and any of the above strategies may be implemented insofar as the methods for diagnosis of disease of the present invention are known as such in the art and constitute further embodiments of the present invention.

[0188] The compounds of the present invention are useful for stratifying patients, i.e., creating subsets within a patient population that provide detailed information about how patients will respond to a given drug. Stratification can be an important component for converting clinical trials that result in negative or neutral results into those with positive results by identifying subsets of the population that are most likely to respond to a new therapy.

[0189] Stratification is the process of identifying patients who share "biological" characteristics to select the optimal management of patients and achieve the best possible outcome in terms of risk assessment, risk prevention, and achieving optimal treatment outcomes. Includes group identification.

[0190] The compounds of the invention can be used to assess or detect as early as possible a particular disease (which is a diagnostic use), the risk of developing the disease (which is a susceptibility / risk use), the progression of the disease, including indolent versus aggressive (which is a prognostic use), and can be used to predict the response and toxicity to a given treatment (which is a predictive use).

[0191] It is also within the scope of the present invention that the compounds of the present invention are used in diagnostic and therapeutic methods.The concept of diagnostic and therapeutic is to combine a therapeutic agent with a corresponding diagnostic test, which can increase the clinical use of the therapeutic agent.The concept of diagnostic and therapeutic is becoming increasingly attractive and is widely considered to be the key to improving the efficiency of drug treatment by identifying patients who will benefit from a given therapy, thus helping doctors to avoid unnecessary treatment.

[0192] The concept of diagnostic therapy is to combine a therapeutic agent with a diagnostic test that allows physicians to identify patients who will benefit most from a given therapy. In an embodiment, and as preferably used herein, the compounds of the present invention are used to diagnose patients, i.e., to identify and localize the primary tumor and potential local and distant metastases. Furthermore, tumor volume can be determined, particularly using three-dimensional diagnostic modalities such as SPECT or PET. Only patients who have FAP-positive tumor masses and therefore will benefit from a given therapy are selected for a specific therapy, thus avoiding unnecessary treatment. Preferably, such therapy is FAP-targeted therapy using the compounds of the present invention. In one particular embodiment, chemically identical tumor-targeted diagnosis, preferably imaging diagnosis for scintigraphy, PET, or SPECT, and radiotherapy are applied. Such compounds differ only in the radionuclide and therefore usually have very similar, if not identical, pharmacokinetic profiles. This can be achieved using chelators and diagnostic or therapeutic radiometals. Alternatively, this can be achieved using precursors for radiolabeling and radiolabeling with diagnostic or therapeutic radionuclides. In one embodiment, diagnostic imaging is preferably used by quantifying the radiation of the diagnostic radionuclide, followed by dosimetry as known to those skilled in the art and predicting drug concentration in the tumor relative to vulnerable side-effect organs. Thus, truly personalized drug administration therapy for the patient is achieved.

[0193] In embodiments, and as preferably used herein, diagnostic therapy is achieved using a single diagnostically and therapeutically active compound, such as a compound of the present invention labeled with a radionuclide that emits diagnostically detectable radiation (e.g., positrons or gamma rays) as well as therapeutically effective radiation (e.g., electrons or alpha particles).

[0194] The present invention also contemplates methods for intraoperatively identifying / disclosing diseased tissue expressing FAP in a subject. Such methods employ compounds of the present invention, whereby such compounds of the present invention preferably include a diagnostically active agent as an effector.

[0195] According to further embodiments of the present invention, the compounds of the present invention, particularly when complexed with radionuclides, are useful in treating many isolated solid cancers, including surgery as a first line method of treatment for many solid cancers; radiation therapy, including the use of ionizing radiation in attempts to cure or ameliorate cancer symptoms using enclosed internal or external sources in the form of brachytherapy; chemotherapy, such as alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antitumor agents; hormonal treatments that modulate the behavior of tumor cells without directly attacking these cells; targeted agents, including monoclonal antibodies and tyrosine kinase inhibitors, that directly target molecular abnormalities in certain types of cancer; angiogenesis inhibitors; immunotherapy; cancer vaccination; and physical, emotional, mental, and psychological treatments to improve the patient's quality of life. It can be used as an adjunct or adjuvant to any other oncology treatment, including palliative care, which includes activities to alleviate social suffering, and alternative treatments, which include a diverse group of products that are not part of the health care system, practice, and conventional medicine.

[0196] In an embodiment of the method of the present invention, the subject is a patient. In an embodiment, the patient is a subject who has been diagnosed with a disease, suspected of having a disease, or is at risk of having or developing a disease, whereby the disease is a disease described herein, preferably a disease including FAP.

[0197] The dosages used in the practice of the methods for treatment and diagnosis, respectively, in which radionuclides are used, more particularly those bound to or part of the compounds of the present invention, will vary depending, for example, on the particular condition to be treated, e.g., the known radiosensitivity of the tumor type, the tumor volume, and the desired therapy. Generally, the dose is calculated based on the radioactivity distribution to each organ and the observed target uptake. The gamma-emitting complex may be administered once or several times for diagnostic imaging. In animals, the indicated dose range is, for example, 1 to 200 MBq. 111 In or 89 The compound of the present invention may be complexed with Zr at 0.1 μg / kg to 5 mg / kg. The beta-emitting complex of the compound of the present invention may be administered at several time points, for example, over a period of 1 to 3 weeks or longer. In animals, the indicated dosage range is, for example, 1 to 200 MBq. 90 Y or 177 In larger animals, e.g., humans, the indicated dosage range is, for example, 10 to 400 MBq. 111 In or 89 In larger animals, e.g., humans, the indicated dosage range is, for example, 10 to 5000 MBq. 90 Y or 177 0.1-100 μg / kg of the compound of the present invention complexed with Lu.

[0198] In a further aspect, the present invention relates, inter alia, to compositions and pharmaceutical compositions comprising the compounds of the present invention. The pharmaceutical composition of the present invention comprises at least one compound of the present invention and, if necessary, one or more carrier substances, excipients and / or adjuvants.The pharmaceutical composition may further comprise, for example, water, a buffer such as neutral buffered saline or phosphate buffered saline, ethanol, mineral oil, vegetable oil, dimethyl sulfoxide, a carbohydrate such as glucose, mannose, sucrose or dextran, mannitol, a protein, an adjuvant, an amino acid such as polypeptide or glycine, an antioxidant, a chelating agent such as EDTA or glutathione, and / or a preservative.In addition, although not required, one or more other active ingredients may be contained in the pharmaceutical composition of the present invention.

[0199] The pharmaceutical composition of the present invention can be formulated for any suitable administration route, including, for example, topical administration such as transdermal or ocular, oral, buccal, nasal, vaginal, rectal or parenteral administration.The term "parenteral" as used herein includes subcutaneous, intradermal, intravascular, such as intravenous, intramuscular, intrathecal and intraperitoneal injection, and any similar injection or infusion technique.The preferred administration route is intravenous administration.

[0200] In an embodiment of the invention, the compounds of the invention containing a radionuclide are administered by any conventional route, in particular intravenously, for example in the form of an injectable solution or suspension. The compounds of the invention may also be advantageously administered by infusion, for example, over a period of 30 to 60 minutes.

[0201] Depending on the location of the tumor, the compounds of the invention can be administered as close to the tumor site as possible, for example, by using a catheter. Such administration can be into the tumor tissue or into the surrounding tissue. The administration of the compounds of the invention can be carried out either directly into the tissue or into the afferent blood vessels. The compounds of the invention can also be administered repeatedly in doses, preferably in divided doses.

[0202] According to a preferred embodiment of the present invention, the pharmaceutical composition of the present invention comprises a stabilizer, such as a free radical scavenger, which inhibits the self-radiolysis of the compound of the present invention. Suitable stabilizers include, for example, serum albumin, ascorbic acid, retinol, gentisic acid or its derivatives, or preferably, electrolyte- and glucose-free amino acid infusion solutions used for parenteral protein supply, such as commercially available amino acid infusion solutions such as Proteinsteril® KE Nephro. Ascorbic acid and gentisic acid are preferred.

[0203] The pharmaceutical compositions of the present invention may contain additional additives, such as agents for adjusting the pH to 7.2-7.4, such as sodium or ammonium acetate or NaHPO. Preferably, a stabilizer is added to the non-radioactive compound of the present invention, and the introduction of the radionuclide, e.g., complexation with the radionuclide, is carried out in the presence of the stabilizer at room temperature or, preferably, at a temperature of 40-120°C. Complexation can be conveniently carried out under air-free conditions, e.g., under N or Ar. Additional stabilizers may be added to the composition after complexation.

[0204] In particular, when the effector is a radionuclide, the excretion of the compounds of the present invention occurs essentially via the kidney.Further protection of the kidney from radioactivity accumulation can be achieved by administering lysine or arginine or an amino acid solution with a high lysine and / or arginine content, such as commercially available amino acid solutions, such as Synthamin®-14 or -10, before or simultaneously with the injection of the compounds of the present invention, especially when the effector is a radionuclide.Kidney protection can also be achieved by administering a plasma expander, such as gelofuscin, instead of or in addition to amino acid infusion.Kidney protection can also be achieved by administering a diuretic, which provides a means of forced diuresis by increasing the rate of urination.Such diuretics include high-sealing loop diuretics, thiazides, carbonic anhydrase inhibitors, potassium-sparing diuretics, calcium-sparing diuretics, osmotic diuretics, and low-sealing diuretics. The pharmaceutical compositions of the present invention may contain, apart from the compounds of the present invention, at least one such further compound intended for or suitable for nephroprotection, preferably nephroprotection of the subject to whom the compounds of the present invention are administered.

[0205] It will be understood by those skilled in the art that the compounds of the present invention are disclosed herein for use in a variety of methods.It will further be understood by those skilled in the art that the compositions of the present invention and the pharmaceutical compositions of the present invention can be used equivalently in the various methods described above.It will also be understood by those skilled in the art that the compositions of the present invention and the pharmaceutical compositions of the present invention are disclosed herein for use in a variety of methods.It will also be understood by those skilled in the art that the compounds of the present invention can be used equivalently in the various methods described above.

[0206] It will be recognized by those skilled in the art that the compositions of the present invention and pharmaceutical compositions of the present invention contain one or more additional compounds in addition to the compounds of the present invention. To the extent that such one or more additional compounds are disclosed herein as being part of the compositions of the present invention and / or pharmaceutical compositions of the present invention, it will be understood that such one or more additional compounds can be administered to the exposed subject or subject of the methods of the present invention separately from the compounds of the present invention. Such administration of one or more additional compounds can be carried out before, simultaneously with, or after the administration of the compounds of the present invention. Also, in the methods of the present invention, one or more additional compounds can be administered separately from the compounds of the present invention. It will be recognized by those skilled in the art that a compound comprising the compound of the present invention may be administered to a subject. Such administration of one or more additional compounds can be carried out before, simultaneously with, or after the administration of the compound of the present invention. To the extent that such one or more additional compounds are disclosed herein as being administered as part of the method of the present invention, it will be understood that such one or more additional compounds are part of the compound of the present invention and / or pharmaceutical composition of the present invention. It is within the scope of the present invention that the compound of the present invention and one or more additional compounds can be contained in the same or different formulations. It is also within the scope of the present invention that the compound of the present invention and one or more additional compounds are not contained in the same formulation, but are contained in the same package containing a first formulation containing the compound of the present invention and a second formulation containing one or more additional compounds, whereby the formulation types can be the same or different.

[0207] It is within the scope of the present invention that more than one type of compound of the present invention be included in the compositions of the present invention and / or pharmaceutical compositions of the present invention, and it is also within the scope of the present invention that more than one type of compound of the present invention be used, and preferably administered, in the methods of the present invention.

[0208] It will be appreciated that the compositions of the invention and pharmaceutical compositions of the invention may be prepared in a conventional manner. Radiopharmaceuticals have a radioactivity content that decreases over time as a result of radioactive decay. The physical half-life of radionuclides is often short for radiopharmaceutical diagnostics. In these cases, final preparation must be carried out immediately before administration to the patient. This is particularly the case for positron-emitting radiopharmaceuticals for tomography (PET radiopharmaceuticals). This often results in the use of semi-finished products such as radionuclide generators, radioactive precursors, and kits.

[0209] Preferably, the kits of the invention will comprise, apart from the one or more compounds of the invention, typically at least one of the following: instructions for use, a final preparation and / or quality control, one or more optional excipients, one or more optional reagents for the labelling procedure, optionally one or more radionuclides with or without a shielded container, and optionally one or more devices, wherein the device is selected from the group comprising a labelling device, a purification device, an analytical device, a handling device, a radiation protection device or an administration device.

[0210] Shielded containers, known as "pig iron" for general handling and transport of radiopharmaceutical containers, come in a variety of configurations to hold radiopharmaceutical containers such as bottles, vials, and syringes. One form often includes a removable cover that allows access to the held radiopharmaceutical solution. With the pig iron cover in place, radiation exposure is tolerable.

[0211] The labeling device is selected from the group of an open reactor, a closed reactor, a microfluidic system, a nanoreactor, a cartridge, a pressure vessel, a vial, a temperature-controllable reactor, a mixing or shaking reactor, and combinations thereof.

[0212] The purification device is preferably selected from the group of an ion exchange chromatography column or device, a size exclusion chromatography column or device, an affinity chromatography column or device, a gas or liquid chromatography column or device, a solid phase extraction column or device, a filtration device, a centrifuge vial column or device.

[0213] The analytical device is preferably selected from the group of test devices for determining the identity, radiochemical purity, radionuclide purity, radioactivity content and specific radioactivity of radiolabeled compounds.

[0214] The handling device is preferably selected from the group consisting of devices for mixing, diluting, dispensing, labeling, injecting and administering radiopharmaceuticals to a subject. Radiation protection devices are used to protect physicians and other individuals from radiation when using therapeutic or diagnostic radionuclides, and are preferably selected from the group consisting of devices having a protective barrier of radiation-absorbing material selected from the group consisting of aluminum, plastic, wood, lead, iron, lead-glass, water, rubber, plastic, and cloth, devices that ensure sufficient distance from the radiation source, devices that reduce exposure time to the radionuclide, devices that limit inhalation, ingestion, or other modes of entry of radioactive material into the body, and devices that provide a combination of these measures.

[0215] The administration device is preferably selected from the group of syringes, syringe guards, needles, pumps, and infusion devices. Syringe guards are generally hollow cylindrical structures that house the cylindrical body of the syringe and are constructed from lead or tungsten containing a lead-glass window that allows the handler to view the syringe plunger and the liquid volume within the syringe.

[0216] The invention will now be further described with reference to the following figures and examples from which further features, embodiments and advantages can be taken. [Brief explanation of the drawings]

[0217] [Figure 1] FIG. 1 shows a radiochromatogram of 177Lu-3BP-3407 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed immediately after synthesis. [Figure 2] FIG. 1 shows a radiochromatogram of 177Lu-3BP-3407 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed 6 days after synthesis. [Figure 3] FIG. 1 shows a radiochromatogram of 177Lu-3BP-3554 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed immediately after synthesis. [Figure 4] FIG. 1 shows a radiochromatogram of 177Lu-3BP-3554 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed 6 days after synthesis. [Figure 5] FIG. 1 shows the percentage of injected dose per gram of tissue (%ID / g) uptake in kidney, liver, blood pool, and HEK-FAP tumor as determined by SPECT imaging of 111In-3BP-3407 1 hour, 3 hours, 6 hours, and 24 hours after injection into a mouse model. [Figure 6] FIG. 1 shows the %ID / g uptake in kidney, liver, blood pool, and HEK-FAP tumor as determined by SPECT imaging of 111In-3BP-3554 1 hour, 3 hours, 6 hours, and 24 hours after injection into a mouse model. [Figure 7] FIG. 1 shows SPECT images of 111In-3BP-3554 1 hour, 3 hours, 6 hours, 24 hours, and 48 hours after injection into mice with HEK-FAP tumors. [Figure 8]FIG. 1 shows the amino acid sequences of human fibroblast activation protein (FAP), human dipeptidyl peptidase 4 (DDP4), and human prolyl endopeptidase (PREP). [Figure 9A] FIG. 1 shows tumor growth over time in HEK-FAP tumor-bearing mice treated with vehicle, non-radioactive compound natLu-3BP-3554, 30 MBq (low dose) of 177Lu-3BP-3554, and 60 MBq (high dose) of 177Lu-3BP-3554. [Figure 9B] FIG. 1 shows the percent body weight change over time in HEK-FAP tumor-bearing mice treated with vehicle, non-radioactive compound natLu-3BP-3554, 30 MBq of 177Lu-3BP-3554, and 60 MBq of 177Lu-3BP-3554. [Figure 10A] FIG. 1 shows representative time-lapse SPECT / CT images of the biodistribution of 60 MBq of 177Lu-3BP-3554 in HEK-FAP tumor-bearing mice. [Figure 10B] FIG. 1 shows representative time-lapse SPECT / CT images of the biodistribution of 30 MBq of 177Lu-3BP-3554 in HEK-FAP tumor-bearing mice. [Figure 11A] FIG. 1 shows representative SPECT / CT images of four different sarcoma PDX models 3 hours after administration of 111In-3BP-3554. [Figure 11B] FIG. 1 shows the %ID / g uptake of 111In-3BP-3554 in four different sarcoma PDX models at 3 hours post-injection. [Figure 12A] FIG. 1 shows tumor growth over time in mice bearing sarcoma Sarc4809 PDX tumors treated with vehicle, non-radioactive compound natLu-3BP-3554, 30 MBq of 177Lu-3BP-3554, or 60 MBq of 177Lu-3BP-3554. [Figure 12B]FIG. 1 shows body weight changes over time in mice bearing sarcoma Sarc4809 PDX tumors treated with vehicle, non-radioactive compound natLu-3BP-3554, 30 MBq of 177Lu-3BP-3554, or 60 MBq of 177Lu-3BP-3554. DETAILED DESCRIPTION OF THE INVENTION

[0218] The following examples are included to provide guidance for those skilled in the art to practice representative embodiments of the presently disclosed subject matter. In view of the present disclosure and the general level of skill in the art, those skilled in the art will recognize that the following examples are intended to be illustrative only, and that numerous changes, modifications, and variations can be employed without departing from the scope of the presently disclosed subject matter. The following synthetic descriptions and specific examples are intended for illustrative purposes only and should not be construed in any way as limiting the ability to prepare compounds of the present disclosure by other methods. [Example]

[0219] Abbreviations used in this application and particularly in the examples below are as follows: 4PL means four parameter logistic curve fitting. Å means angstrom.

[0220] ACN means acetonitrile. Ahx means 6-aminohexanoic acid. AMC means 7-amino-4-methylcoumarin.

[0221] amu means atomic mass unit. aq. means aqueous. AUC inf means the area under the curve extrapolated to infinity.

[0222] BSA means bovine serum albumin. C0 denotes the initial concentration of the compound. CAF means cancer-associated fibroblasts.

[0223] CL means clearance. CM means ChemMatrix™. CT stands for computed tomography.

[0224] Cy5 means cyanine-5. DAD stands for Diode Array Detector. DCM means dichloromethane.

[0225] Dde means N-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl). DEG means diethylene glycol dimethacrylate.

[0226] DIC means N,N'-diisopropylcarbodiimide. DICOM stands for Digital Imaging and Communications in Medicine.

[0227] DIPEA means diisopropylethylamine. DMF means N,N-dimethylformamide. DMSO means dimethyl sulfoxide.

[0228] DOTA means 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid. DOTA(tBu)3-OH means tri-tert-butyl-1,4,7,10-tetraazacyclo-dodecane-1,4,7,10-tetraacetate.

[0229] DPP means dipeptidyl peptidase. EC means electron capture. EC 50 means half-maximal excitation concentration.

[0230] ECACC stands for European Collection of Authenticated Cell Cultures. EDC means 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide.

[0231] EMEM means Eagle's minimum essential medium. eq or eq. means equivalent. ESI means electrospray ionization.

[0232] Et2O means diethyl ether. EtOAc means ethyl acetate. FACS stands for fluorescence activated cell sorting.

[0233] FAP stands for fibroblast activation protein. Fb means background fluorescence intensity. FBS means fetal bovine serum.

[0234] FGF21 means fibroblast growth factor 21. FITC means 5(6)-fluorescein isothiocyanate. Fmoc means 9-fluorenylmethoxycarbonyl.

[0235] FRET stands for Fluorescence Resonance Energy Transfer. Ft means fluorescence intensity. Gab means gamma-aminobutyric acid.

[0236] GABA means gamma-aminobutyric acid. h means hours. HATU means O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0237] HBST means SPR running buffer. HEK-FAP means human embryonic kidney 293 cells expressing human FAP. HEPES means 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid.

[0238] HFIP means hexafluoro-2-isopanol. HOAc means acetic acid. HOAt means 1-hydroxy-7-azabenzotriazole.

[0239] HPLC means high performance liquid chromatography. HPLC / MS means high performance liquid chromatography / mass spectrometry. I C 50 means half-maximal inhibitory concentration.

[0240] ID / g means injected dose per gram. IS means nuclear isomer transition. iTLC-SG means instant thin layer chromatography-silica gel.

[0241] K2EDTA means dipotassium ethylenediaminetetraacetic acid. K D means the dissociation constant. kDa means 1000 Daltons.

[0242] K i means the inhibition constant. k off means the dissociation rate. k on means the association rate.

[0243] LC / TOF-MS stands for liquid chromatography / time of flight / mass spectrometry. LC-MS means high performance liquid chromatography with mass spectrometry. LDH means lactate dehydrogenase.

[0244] Leu means leucine. LiOH means lithium hydroxide. M means molar concentration or moles per liter.

[0245] m / z means mass divided by charge. max. means maximum. MeOH means methanol.

[0246] MeV stands for megaelectronvolt. min means minutes. MMP means matrix metalloproteinase.

[0247] MRM stands for multiple reaction monitoring. MTBE means methyl tert-butyl ether. Mtt means methyltrityl.

[0248] MTV means mean tumor volume. MW means molecular weight. nd means undecided.

[0249] Na2SO4 means sodium sulfate. NaCl means sodium chloride. NaHCO3 means sodium bicarbonate.

[0250] NCA stands for non-compartmental analysis. NHS means N-hydroxysuccinimide. NMP means 1-methyl-2-pyrrolidone.

[0251] NOS means not specified. Oic means L-octahydroindole-2-carboxylic acid. pa means for analytical purposes (quality grade).

[0252] pi means post injection. Pbf means 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-sulfonyl

[0253] PBS means phosphate buffered saline. PDX means patient-derived xenograft. PET stands for positron emission tomography.

[0254] pIC50 means the negative logarithm of the IC50 value when converted to a molar concentration. POP means prolyl oligopeptidase. ppm means parts per million.

[0255] PREP means prolyl endopeptidase. Prep. means to separate. PS means polystyrene.

[0256] Q-TOF stands for quadrupole time of flight. Ref means reference. RFU means relative fluorescence units.

[0257] RLB means radioligand binding assay. RMCE stands for recombinase-mediated cassette exchange. RP means reverse phase.

[0258] R t means retention time. RT means room temperature. RU means resonance unit.

[0259] SAR stands for structure-activity relationship. sat. means saturated. SCID stands for severe combined immunodeficiency disorder.

[0260] SCK stands for single cycle kinetics. sec or s means seconds. SF stands for spontaneous fission.

[0261] SPECT stands for single photon emission computed tomography. SPPS stands for solid phase peptide synthesis. t 1 / 2 means terminal half-life.

[0262] tBu means tert.butyl. TFA means trifluoroacetate or trifluoroacetic acid. TG means TentaGel.

[0263] TGI means tumor growth inhibition. THF means tetrahydrofuran. TIPS means triisopropylsilane.

[0264] TLC means thin layer chromatography. TME means tumor microenvironment. t R means retention time.

[0265] UHPLC stands for ultra-high performance liquid chromatography. UV means ultraviolet. V ss means the volume of distribution at steady state.

[0266] V Z means the volume of distribution in the terminal phase. Example 1 material and method The materials and methods, as well as general procedures, are further illustrated by the following examples.

[0267] solvent: Solvents were used in the specified quality without further purification. Acetonitrile (Super Gradient, HPLC, VWR - for analytical purposes; PrepSolv, Merck - for preparative purposes); dichloromethane (synthetic, Roth); ethyl acetate (synthetic grade, Roth); N,N-dimethylformamide (peptide synthesis grade, Biosolve); 1-methyl-2-pyrrolidone (peptide grade, IRIS BioTech); 1,4-dioxane (reinst, Roth); methanol (pa, Merck).

[0268] Water: Milli-Q Plus, Millipore, desalted. Chemicals: Chemicals were synthesized according to literature procedures or similarly synthesized to literature procedures or commercially available from Sigma-Aldrich-Merck (Deisenhofen, Germany), Bachem (Bubendorf, Switzerland), VWR (Darmstadt, Germany), Novabiochem (Merck Group, Darmstadt, Germany), Acros Organics (distributor Fisher Scientific GmbH, Schwerte, Germany), Iris Biotech (Marktredwitz, Germany), Amatek Chemical (Jiangsu, China), Roth (Karlsruhe, Germany), and others. , Germany), Molecular Devices (Chicago, USA), Biochrom (Berlin, Germany), Peptech (Cambridge, MA, USA), Synthetech (Albany, OR, USA), Pharmacore (High Point, NC, USA), PCAS Biomatrix Inc. (Saint-Jean-sur-Richelieu, Quebec, Canada), Alfa Aesar (Karlsruhe, Germany), Tianjin Nankai Hecheng S&T Co., Ltd. (Tianjin, China), CheMatech (Dijon, France), and Anaspec (San Jose, CA, USA), or other companies, and were used at the specified quality without further purification.

[0269] cell: HT29 (ECACC catalog number 91072201) and WI-38 (ECACC catalog number 90020107) were purchased from ECACC, and HEK293 cells (Q12884) expressing human FAP were produced by InSCREENeX GmbH (Braunschweig, Germany) using recombinase-mediated cassette exchange (RMCE), as described by Nehlsen et al. (2009, 9:100). HPLC / MS analysis HPLC / MS analysis was performed by injecting 5 μl of sample solution and using a two-step gradient (5 to 65% B in 12 min, followed by 65 to 90% in 0.5 min, A: 0.1% TFA in water, and B: 0.1% TFA in ACN) for all chromatograms. The RP column was from Agilent (Type Poroshell 120, 2.7 μm, EC-C18, 50 × 3.00 mm, flow rate 0.8 ml, HPLC at room temperature); mass spectrometer: Agilent 6230 LC / TOF-MS, ESI ionization. MassHunter Qualitative Analysis B.07.00 SP2 was used as software. UV detection was performed at λ = 230 nm. Retention time (R t ) are expressed in decimal notation (e.g., 1.9 min = 1 min 54 s) and refer to detection by UV spectrometer. The "Find Compounds by Formula" function was used to assess the masses of the observed compounds. Specifically, the individual "Compound Neutral Mass (unit: Daltons)" values ​​and the corresponding isotope distribution patterns were used to confirm the identity of the compounds. The mass spectrometer precision was approximately ±5 ppm.

[0270] Preparative HPLC: Preparative HPLC separations were performed using reverse-phase columns (Kinetex 5μ XB-C18 100Å, 150 × 30 mm, or RLRP-S 8μ, 100Å, 150 × 25 mm, manufactured by Phenomenex). The mobile phases were 0.1% TFA in water (A) and 0.1% TFA in ACN (B), mixed in a linear binary gradient. This gradient is described as "10 to 40% in 30 min," meaning a linear gradient from 10% B (and corresponding 90% A) to 40% B (and corresponding 60% A) was performed within 30 min. The flow rate was in the range of 30–50 ml / min. A typical gradient for the purification of compounds of the present invention began with 5–25% B and ended after 30 min at 30–50% B, with a difference in the percentage of B between the end and start points of at least 10%. A commonly used gradient was 15 to 40% B in 30 minutes.

[0271] General procedure for automated / semi-automated solid phase synthesis: Automated solid phase synthesis of peptides and polyamides was carried out on a Tetras Peptide Synthesizer (Advanced ChemTech) at the 50 μmol and 100 μmol scales. Manual steps were performed using a plastic slide with a frit. The reactions were carried out in syringes (material PE, Roland Vetter Laborbedarf OHG, Ammerbuch, Germany). The amounts of reagents in the described protocols correspond to a 100 μmol scale unless otherwise stated.

[0272] Solid-phase synthesis was performed on polystyrene (crosslinked with 1,4-divinylbenzene (PS) or di(ethylene glycol) dimethacrylate (DEG)), ChemMatrix (CM), or TentaGel (TG) resin. Resin linkers were trityl, wang, and rink amide.

[0273] Resin filling: In the case of the trityl linker, attachment of the first building block (resin loading) was performed as follows: The resin (polystyrene (PS) trityl chloride, initial loading: 1.8 mmol / g) was swollen in DCM (5 ml) for 30 min, followed by washing with DCM (3 ml, 1 min). The resin was then treated with a mixture of the corresponding building block (0.5 mmol, 5 eq.) and DIPEA (350 μl, 3.5 mmol, 35 eq.) in DCM (4 ml) for 1 h. The resin was then washed with methanol (5 ml, 5 min) and DMF (3 ml, 2 x 1 min).

[0274] In the case of the Wang linker, pre-loaded resins (polystyrene (PS) and TentaGel (TG)) were used. In the case of the Rink Amide linker, attachment of the first residue to the resin (CM, DEG) was carried out by the same procedure as for chain assembly described below.

[0275] Alloc / Allyl Deprotection: After swelling in DMF, the resin was washed with DMF and DCM. The DCM was deoxygenated by passing a stream of nitrogen through the stirred solvent. The resin was washed twice with oxygen-free solvent. 2 ml of a 2 M solution of barbituric acid in oxygen-free DCM and 1 ml of a 25 μM solution of tetrakis(triphenylphosphine)palladium(0) in oxygen-free DCM were then added to the resin. The resin was stirred for 1 hour and then washed with DCM, MeOH, DMF, 5% DIPEA in DMF, 5% dithiocarbamate in DMF, DMF, and DCM (each washing step was repeated three times for 3 ml and 1 min).

[0276] Fmoc deprotection: After swelling in DMF, the resin was washed with DMF and then treated with piperidine / DMF (1:4, 3 ml, 2 and 20 min), followed by washing with DMF (3 ml, 5 times x 1 min).

[0277] Dde deprotection: After swelling in DMF, the resin was washed with DMF and then treated with hydrazine hydrate / DMF (2 / 98, 3 ml, 2 times 10 min) followed by washing with DMF (3 ml, 5 times 1 min).

[0278] Mtt deprotection: After swelling in DCM, the resin was washed with DCM and then treated with HFIP / DCM (7 / 3, 4–6 ml, 4 h), followed by washing with DCM (3 ml, 3 times × 1 min), DMF (3 ml, 3 times × 1 ml), and DIPEA (0.9 M in DMF, 3 ml, 1 min).

[0279] Reagent solutions: Building blocks (0.3M in DMF or NMP), DIPEA (0.9M in DMF), HATU (0.4M in DMF), acetic anhydride (0.75M in DMF) Coupling: Coupling of building blocks / amino acids (chain construction): Unless otherwise stated, coupling of building blocks was carried out as follows: After subsequent addition of a solution of the corresponding building block (1.7 mL, 5 eq.), a DIPEA solution (1.15 ml, 10 eq.), and a HATU solution (1.25 ml, 5 eq.), the resin was shaken for 45 min. If necessary, the resin was washed with DMF (3 ml, 1 min) and the coupling step was repeated.

[0280] Terminal acetylation: After addition of DIPEA solution (1.75 ml, 16 eq.) and acetic anhydride solution (1.75 ml, 13 eq.), the resin was shaken for 10 min, after which it was washed with DMF (3 ml, 6 times 1 min).

[0281] Cleavage Method A: Cleavage of Protected Fragments from Highly Acid-Labile Resins: After completion of the sequence assembly, the resin was finally washed with DCM (3 ml, 4 times 1 min) and then dried in vacuo. The resin was then treated with HFIP / DCM (7 / 1, 4 ml, 4 h), and the collected solution was evaporated to dryness. The residue was either purified by preparative HPLC or used without further purification.

[0282] Cleavage Method B: Cleavage of the Unprotected Fragment (Full Resin Cleavage): After completion of the sequence assembly, the resin was finally washed with DCM (3 ml, 4 times 1 min), dried in vacuo overnight, and (unless otherwise stated) treated with TFA, EDT, water, and TIPS (94 / 2.5 / 2.5 / 1) for 2 h. The cleavage solution was then poured into a cold mixture of MTBE and cyclohexane (1 / 1, 10-fold excess relative to the volume of the cleavage solution), centrifuged for 5 min at 4 °C, and the precipitate was collected and dried in vacuo. The residue was lyophilized from water / acetonitrile prior to purification or further modification.

[0283] Cleavage Method C: Cleavage of Peptide Protecting Groups in Solution The protected / partially protected compounds were dissolved in THF, water, and TIPS (95 / 2.5 / 2.5) for 2 hours (unless otherwise stated). The cleavage solution was then poured into a cold mixture of MTBE and cyclohexane (1 / 1, 10-fold excess relative to the volume of the cleavage solution), centrifuged at 4°C for 5 minutes, and the precipitate was collected and dried in vacuo. The residue was lyophilized from water / acetonitrile prior to purification or further modification.

[0284] A more suitable Fmoc solid phase peptide synthesis method is described in detail in "Fmoc Solid Phase Peptide Synthesis" Editors W. Chan, P. White, Oxford University Press, USA, 2000. Compounds were named using MestreNova version 12 Mnova IUPAC Name plug-in (Mestrelab Research, SL) or AutoNom version 2.2 (Beilstein Informationssysteme Copyright 1988-1998, Beilstein Institut für Literatur der Organischen Chemie licensed to Beilstein Chemiedaten and Software GmbH), as appropriate.

[0285] Compound preparation: Specific embodiments of the preparation of compounds of this invention are provided in the Examples below. Unless otherwise specified, all starting materials and reagents are of standard commercial grade and used without further purification, or are readily prepared from such materials by conventional methods. Those skilled in the art of organic synthesis will readily appreciate the methods used to prepare compounds encompassed by this invention. It will be recognized in light of this disclosure that the starting materials and reaction conditions may be varied, including additional steps.

[0286] One general synthetic route to the compounds of the invention involves: 1. Solid-phase peptide synthesis (SPPS) of a linear peptide precursor bearing two thiol moieties.

[0287] 2. Thiol-site-directed cyclization of this linear peptide precursor by: a. Bis(bromomethyl)benzene derivatives or b. Tris(bromomethyl)benzene derivatives.

[0288] 3. In the case of cyclization with tris(bromomethyl)benzene derivatives, the intermediate formed in this cyclization reaction was further reacted with a linker that allows the attachment of a chelating agent. Example 2 Synthesis of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554) The title compounds were synthesized by first synthesizing a linear peptide precursor on the solid phase, followed by solution phase cyclization (Example 2a, in either a non-aqueous solution (Method A) or an aqueous solution (Method B)), or alternatively, by performing all steps, including solid phase cyclization, on the solid phase (Example 2b).

[0289] Example 2a Synthesis in solution by two alternative cyclization methods As described in "General Procedure for Automated / Semi-Automated Solid-Phase Synthesis," Fmoc-Cys(Trt)-OH was loaded onto a trityl resin on a 50 μmol scale. A peptide of the sequence (Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-OH) was assembled onto this resin according to "General Procedure for Automated / Semi-Automated Solid-Phase Synthesis." After performing the "Cleavage Method B" step, the crude peptide was lyophilized and cyclized in solution by two alternative methods.

[0290] Cyclization method A: The crude peptide (based on 50 μmol resin loading) was dissolved in 10 ml of a 1:1 mixture of ethanol and acetonitrile. To this mixture, 35 μl of DIPEA was first added, followed by 23.7 mg of 1,3,5-tris(bromomethyl)benzene (66.6 μmol, 1.3 eq relative to the initial resin loading). The solution was stirred for 1 h, and then 42.8 mg of cysteamine (555 μmol, 11 eq relative to the initial resin loading) was added. After 1 h, the solvent was removed in vacuo, and 25 ml of a 1:1 mixture of acetonitrile and water (containing 50 μl of TFA) was added. The solvent was removed by lyophilization. The residue was subjected to HPLC purification (15 to 45% B-Kinetex in 30 min) to give 17.8 mg (16.4 μmol) (32.8%) of the intermediate Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH.

[0291] Cyclization method B: The crude peptide (based on 50 μmol resin loading) was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH = 8.5) and acetonitrile. To this mixture was added 26.8 mg (75 μmol, 1.5 eq. relative to the initial resin loading) of 1,3,5-tris(bromomethyl)benzene in 0.5 ml of acetonitrile. The solution was stirred for 1 hour, and then 38.6 mg (500 μmol) of cysteamine was added. mol, 10 eq relative to the initial resin loading. After 2 h, 50 μl of TFA was added, and the solvent was removed by lyophilization. The residue was subjected to HPLC purification (15 to 45% in 30 min B-Kinetex) to give 19.47 mg (18 μmol) of the intermediate Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (35.9%).

[0292] Both cyclization methods performed similarly, achieving comparable yields and similar purities. To a solution of the intermediate Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (obtained in this example by cyclization method B) in 300 μl of DMSO, 5 μl of DIPEA was added to adjust the pH to approximately 7.5-8. Then, 20.5 mg of DOTA-NHS (27 μmol, 1.5 equivalents relative to the peptide intermediate) in 200 μl of DMSO was added. During the course of the reaction, monitored by LC / TOF-MS, 5 μl of DIPEA was added three times to readjust the pH to the starting value. After completion of the reaction, the solution was subjected to HPLC purification (15-45% in 30 min B-Kinetex) to obtain 20.44 mg of pure title compound (27.8% overall yield). HPLC:R t = 5.9 min. LC / TOF-MS: Exact mass 1469.640 (calculated 1469.639). 67 H 99 N 13 O 18 S3 (MW=1470.780).

[0293] Example 2b Synthesis including solid phase cyclization For the synthesis of the resin-bound title compound, Fmoc-Cys(Trt)-WANG Tentagel resin was used as the starting material. A peptide with the sequence (Hex-Cys(Trt)-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys-OH) was assembled on this resin on a 1 mmol scale according to the "General Procedure for Automated / Semi-Automated Solid-Phase Synthesis." After completion of the assembly, the resin was washed with DCM (3 times 1 min). The trityl protecting group was then selectively removed from the resin by treatment with a solution of TFA, TIPS, and DCM (5 / 5 / 90, 5 x 5 min). The resin was washed with DCM, DMF, 0.9 M DIPEA in DMF, DMF, DCM (3 / 3 / 2 / 3 / 3), and dried in vacuo. The following cyclization was performed in 200 μmol portions. For this purpose, the resin was swollen in DMF and then treated with a solution of 1,3,5-tris(bromomethyl)benzene (86 mg, 240 μmol, 1.2 eq), DIPEA (235 μL, 1 mmol, 5 eq) in 2 mL of DMF at 50° C. for 90 min. The solution was removed, the resin was washed with DMF, and then a solution of cysteamine (154.3 mg, 2 mmol, 10 eq) was added to the resin. The resin was stirred for another 90 min at 50° C. After washing the resin with DMF and DCM (3 / 3), the peptide resin (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) was dried. This procedure may result in partial or complete deprotection of the trityl group on glutamine. In any case, this does not prevent the optional derivatization of the free amino groups of the AET.

[0294] For the final derivatization with DOTA, peptide resin (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) was used on a 50 μmol scale. DOTA(tBu)3-OH was coupled according to the "General Procedure for Automated / Semi-Automated Solid-Phase Synthesis." After drying, the resin was subjected to "Cleavage Method B." The crude peptide was lyophilized and subsequently purified by preparative HPLC (15 to 45% B in 30 min - Kinetex) to yield 11.0 mg (7.5 μmol) of pure title compound (15%). HPLC:R t = 5.9 min. LC / TOF-MS: Exact mass 1469.640 (calculated 1469.639). 67 H 99 N 13 O 18 S3 (MW=1470.780).

[0295] Example 3 Synthesis of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407) a) Synthesis of the intermediate Hex-[Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 by two different cyclization methods The peptide of the sequence (Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-Asp-NH) was assembled on Rink amide resin at a 50 μmol scale according to the "General Procedure for Automated / Semi-Automated Solid-Phase Synthesis." After performing the "Cleavage Method B" step, the crude peptide was lyophilized and cyclized by two alternative methods.

[0296] Cyclization method A: The crude peptide (based on 50 μmol resin loading) was dissolved in 10 ml of a 1:1 mixture of ethanol and acetonitrile. To this mixture, 30 μl of DIPEA was first added, followed by 26.8 mg (75 μmol, 1.5 eq. relative to the initial resin loading) of 1,3,5-tris(bromomethyl)benzene. The solution was stirred for 45 min, and then piperazine in 200 μl of a 1:1 mixture of ethanol / acetonitrile was added. A solution of 43 mg (500 μmol, 10 eq relative to the initial resin loading) of acetonitrile was added. After 1 h, the solvent was removed in vacuo, and 25 ml of a 1:1 mixture of acetonitrile and water (containing 50 μl of TFA) was added, followed by lyophilization to remove the solvent. The residue was subjected to HPLC purification (15 to 40% in 30 min B-Kinetex) to yield 15.3 mg (12.7 μmol) (25.3%) of the peptide intermediate Hex-Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH

[0297] Cyclization method B: The crude peptide (based on a 50 μmol resin loading) was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH = 8.5) and acetonitrile. To this mixture, 26.8 mg (75 μmol, 1.5 eq relative to the initial resin loading) of 1,3,5-tris(bromomethyl)benzene was added. The solution was stirred for 1 h, and 43 mg (500 μmol, 10 eq relative to the initial resin loading) of piperazine was added. After 6 h, 100 μl of TFA was added, and the solvent was removed by lyophilization. The residue was subjected to HPLC purification (15 to 40% in 30 min B-Kinetex) to yield 17.2 mg (14.2 μmol) (28.4%) of the peptide intermediate Hex-Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2.

[0298] The performance of both cyclization methods is similar, achieving comparable yields and purities. b) Final steps in the synthesis of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407): DOTA coupling and purification To a solution of the intermediate (obtained by cyclization method B) in 200 μl of DMSO, 2.5 μl of DIPEA was added to adjust the pH value to approximately 7.5-8. Then, 16.3 mg (21.4 μmol, 1.5 eq. relative to the peptide intermediate) of DOTA-NHS in 100 μl of DMSO was added. During the course of the reaction, monitored by LC / TOF-MS, 2.5 μl of DIPEA was added five times to readjust the pH value to the starting value. After completion of the reaction, the solution was subjected to HPLC purification (15 to 40% B in 30 min - Kinetex) to give 19.1 mg (12.0 μmol) (85%) of the pure title compound. HPLC:R t = 5.70 min. LC / TOF-MS: Exact mass 1592.737 (calculated 1592.737). 73 H 108 N 16 O 20 S2 (MW = 1593.866).

[0299] Example 4 Preparation of DOTA-Transition Metal Complexes of Compounds of the Invention A. General procedure for the preparation of peptides containing DOTA-transition metal complexes from the corresponding peptides containing uncomplexed DOTA. in 0.4 M sodium acetate, pH = 5 (Buffer A) (for Cu(II), Zn(II), In(III), Lu(III), or Ga(III) complexes) or in 0.1 M ammonium acetate, pH = 8 (buffer B) (for Eu(III) complexes) A 0.1 mM solution of the peptide composed of uncomplexed DOTA was diluted with a 0.1 mM solution of the corresponding metal salt in water, thereby adjusting the molar ratio of peptide to metal to 1:3. Stirred for 20 minutes at 50°C (also referred to herein as condition A) (for In(III), Lu(III), Ga(III), Zn(II), or Cu(II) complexes), or or Overnight at room temperature (also referred to herein as condition B) (for Eu(III) complexes) The mixture was stirred at RT.

[0300] This solution is then applied to HPLC purification (also referred to herein as Purification A), or Solid phase extraction (also referred to herein as purification B) was applied. For solid-phase extraction, 250 mg of Varian Bondesil-ENV was placed in a 15 ml polystyrene syringe and pre-washed with methanol (1 x 5 ml) and water (2 x 5 ml). The reaction solution was then applied to the column. Elution was then performed with water (2 x 5 ml - to remove excess salts), 5 ml of 50% ACN in water as the first fraction, and each subsequent fraction was eluted with 5 ml of 50% ACN in water containing 0.1% TFA.

[0301] In both cases (HPLC purification or solid phase extraction), fractions containing pure product were pooled and lyophilized. B. Indium complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3590) The conjugate was prepared starting from 25 mg (15.7 μmol) of peptide 3BP-3407 dissolved in buffer A, diluted with a solution of InCl3×4H2O, and treated under condition A. For the purification step, "Purification A" was used (15 to 40% B-RLRP-S in 30 min) to give 18.24 mg (68.1% yield) of the pure title compound. HPLC:R t = 5.6 min. LC / TOF-MS: Exact mass 1702.622 (calculated 1702.617). 73 H 105 InN 16 O20 S2 (MW=1705.663).

[0302] C. Gallium complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3592) The conjugate was prepared starting from 25 mg (15.7 μmol) of peptide 3BP-3407 dissolved in buffer A, diluted with a solution of Ga(NO3)3 × H2O, and treated under condition A. For the purification step, "Purification A" was used (15 to 40% B-RLRP-S in 30 min) to give 16.78 mg (69.3% yield) of the pure title compound. HPLC:R t = 5.7 min. LC / TOF-MS: Exact mass 1658.664 (calculated 1658.639). 73 H 105 GaN 16 O 20 S2 (MW=1660.568).

[0303] D. Lutetium complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3591) The conjugate was prepared starting from 25 mg (15.7 μmol) of peptide 3BP-3407 dissolved in buffer A, diluted with a solution of LuCl3, and treated under condition A. For the purification step, "Purification A" was used (15 to 40% B-RLRP-S in 30 min) to give 16.66 mg (60.1% yield) of the pure title compound. HPLC:R t = 5.6 min. LC / TOF-MS: Exact mass 1764.654 (calculated 1764.654). 73 H 105 LuN 16 O 20 S2 (MW=1765.812).

[0304] E. Europium complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3661) The conjugate was prepared starting from 9.5 mg (6 μmol) of peptide 3BP-3407 dissolved in buffer B, diluted with a solution of EuCl3×6H2O, and treated under condition B. In the purification step, "Purification B" was used to obtain 8.24 mg (79.3% yield) of the pure title compound. HPLC: t = 5.7 min. LC / TOF-MS: Exact mass 1740.636 (calculated 1740.633). 73 H 105 EuN 16 O 20 S2 (MW=1742.809).

[0305] F. Indium complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3623) The conjugate was prepared starting from 6 mg (4.1 μmol) of peptide 3BP-3554 dissolved in buffer A, diluted with a solution of InCl3×4H2O, and treated under condition A. For the purification step, "Purification B" was used to obtain 5.26 mg (81% yield) of the pure title compound. HPLC:R t = 5.8 min. LC / TOF-MS: Exact mass 1579.524 (calculated 1579.520). 67 H 96 InN 13 O 18 S3 (MW=1582.574).

[0306] G. Lutetium complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3624) The conjugate was prepared starting from 6 mg (4.1 μmol) of peptide 3BP-3554 dissolved in buffer A, diluted with a solution of LuCl3, and treated under condition A. In the purification step, "Purification B" was used to obtain 5.5 mg (82% yield) of the pure title compound. HPLC: t = 5.9 min. LC / TOF-MS: Exact mass 1641.560 (calculated 1641.557). 67 H 96 LuN 13 O18 S3 (MW=1642.723).

[0307] H. Gallium complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3949) The conjugate was prepared starting from 7.9 mg (5.4 μmol) of peptide 3BP-3554 dissolved in buffer A, diluted with a solution of Ga(NO3)3×H2O, and treated under condition A. For the purification step, "Purification B" was used to obtain 4.2 mg (51% yield) of the pure title compound. HPLC:R t = 6.6 min. LC / TOF-MS: Exact mass 1535.543 (calculated 1535.541). 67 H 96 GaN 13 O 18 S3 (MW=1537.479).

[0308] I. Europium complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3662) The conjugate was prepared starting from 3.4 mg (2.3 μmol) of peptide 3BP-3554 dissolved in buffer B, diluted with a solution of EuCl3×6H2O, and treated with condition B. In the purification step, "Purification B" was used to obtain 3.1 mg (83% yield) of the pure title compound. HPLC: t = 5.9 min. LC / TOF-MS: Exact mass 1617.54 1 (calculated value 1617.536).C 67 H 96 EuN 13 O 18 S3 (MW=1619.721).

[0309] J. Copper(II) complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4293) The conjugate was prepared starting from 18 mg (12.2 μmol) of peptide 3BP-3554 dissolved in buffer A, diluted with a solution of Cu(OAc)2, and treated under condition A. For the purification step, "Purification B" was used to obtain 16.5 mg (88% yield) of the pure title compound. HPLC: t = 6.5 min. LC / TOF-MS: Exact mass 1530.553 (calculated 1530.553). 67 H 97 CuN 13 O 18 S3 (MW=1532.310).

[0310] K. Zinc complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4343) The conjugate was prepared starting from 20 mg (13.6 μmol) of peptide 3BP-3554 dissolved in buffer A, diluted with a solution of ZnCl2, and treated under condition A. For the purification step, "Purification B" was used to obtain 16.1 mg (77% yield) of the pure title compound. HPLC: t = 6.4 min. LC / TOF-MS: Exact mass 1531.553 (calculated 1531.553). 67 H 97 N 13 O 18 S3Zn (MW = 1534.160).

[0311] Example 5 Plasma stability assay Plasma stability assays were performed to determine the stability of selected compounds of the present invention in human and mouse plasma. Such plasma stability assays measure the degradation of compounds of the present invention in plasma. This is an important characteristic of the compounds, since compounds that are rapidly degraded in plasma, with the exception of prodrugs, generally exhibit low efficacy in vivo. The results indicate that these compounds are highly stable in human and mouse plasma. This stability is sufficient for the diagnostic, therapeutic, and diagnostic-therapeutic uses of these compounds of the present invention.

[0312] Plasma stability samples were prepared by mixing 50 μl of plasma aliquots (all K2EDTA) with 1 μl of 0.5 mM compound stock solution in DMSO. After vortexing, samples were incubated in a Thermomixer at 37°C for 0, 4, and 24 hours. After incubation, samples were stored on ice until further processing. All samples were prepared in duplicate.

[0313] The appropriate internal standard was added to each sample (1 μl of a 0.5 mM stock solution in DMSO). Protein precipitation was performed using two different methods depending on the compound conditions shown in Table 5:

[0314] A) 250 μl of acetonitrile containing 1% trifluoroacetic acid was added. After 30 minutes of incubation at room temperature, the precipitate was separated by centrifugation and 150 μl of the supernatant was diluted with 150 μl of 1% aqueous formic acid.

[0315] B) 150 μl of zinc sulfate precipitant containing 78% 0.1 M zinc sulfate and 22% acetonitrile was added. After 30 minutes of incubation at room temperature, the precipitate was separated by centrifugation. If the compound contained a free DOTA moiety, 1% acetonitrile was added to 100 μl of the supernatant. The formation of the zinc chelate was completed by adding 10 μl of formic acid followed by further incubation at 60° C. for 10 minutes.

[0316] Analyte determination in clean sample solutions was performed on an Agilent 1290 UHPLC system coupled with an Agilent 6530 Q-TOF mass spectrometer. Chromatographic separation was performed on a Phenomenex BioZen XB-C18 HPLC column (50 × 2 mm, 1.7 μm particle size) by gradient elution using a mixture of 0.1% formic acid in water as eluent A and acetonitrile as eluent B (2% B to 41% in 7 min, 800 μl / min, 40 °C). Mass spectrometric detection was performed in positive ion ESI mode by scanning the mass range from m / z 50 to 3000 at a sampling rate of 2 / s.

[0317] From the raw mass spectrometry data, the ion currents of doubly or triply charged monoisotopic signals were extracted for both the compounds and the internal standards. Quantitation was performed by external matrix calibration with an internal standard using integrated analyte signals.

[0318] In addition, recovery was determined by spiking pure plasma samples containing only the internal standard after treatment with a specific amount of compound. Carryover was assessed by analysis of a blank sample (20% acetonitrile) after the highest calibration sample.

[0319] The results of this assay, performed on some of the compounds of the present invention, are shown in Table 5 below. Results are presented as "% intact compound remaining after 24 hours," meaning that from the amount of material at the start of the experiment, the indicated percentage was detected as unchanged material by LC-MS quantification at the end of the experiment. All compounds were greater than 50% intact after at least 24 hours, indicating that these compounds are considered to be sufficiently stable for diagnostic and therapeutic applications.

[0320] [Table 5]

[0321] Example 6 FACS binding assay To determine the binding of compounds of the present invention to FAP-expressing cells, a competitive FACS binding assay was established.

[0322] FAP-expressing human WI-38 fibroblasts (FCACC) were cultured in EMEM containing 15% fetal bovine serum, 2 mM L-glutamine, and 1% non-essential amino acids. Cells were detached with Accutase (Biolegend, #BLD-423201) and washed with FACS buffer (PBS containing 1% FBS). Cells were diluted with FACS buffer to a final concentration of 100,000 cells per ml, and 200 μl of the cell suspension was transferred to a U-shaped tube. The cells were transferred to a non-binding 96-well plate (Greiner). The cells were washed with ice-cold FACS buffer and incubated with 3 nM of Cy5-labeled compound (H-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-His-Phe-Arg-Asp-Ttds-Lys(Cy5SO3)-NH2) along with increasing concentrations of peptide for 1 h at 4 °C. The cells were washed twice with FSCS buffer and resuspended in 200 μl of FACS buffer. The cells were analyzed using an Attune NxT flow cytometer. The median fluorescence intensity (Cy5 channel) was calculated using the Attune NxT software and plotted against peptide concentration. Four-parameter logistic (4PL) curve fitting and pIC50 calculation were performed using ActivityBase software. The results of this assay and the FAP protease activity assay of Example 7 for each compound of the invention are shown in Table 6 (shown in Example 7). pIC50 Category A represents pIC50 values ​​greater than 8.0, Category B represents pIC50 values ​​between 7.1 and 8.0, Category C represents pIC50 values ​​between 6.1 and 7.0, and Category D represents pIC50 values ​​of 6.0 or less.

[0323] Example 7 FAP protease activity assay To determine the inhibitory activity of the peptides of Example 6, a FRET-based FAP protease activity assay was established.

[0324] Recombinant human FAP (R&D systems, #3715-SE) was diluted to a concentration of 3.6 nM in assay buffer (50 mM Tris, 1 M NaCl, 1 mg / mL BSA, pH 7.5). 25 μl of this FAP solution and 3-fold serial dilutions of test compounds were added. The mixture was mixed with 25 μl of FRET-peptide HiLyteFluor™ 488-VS(D-)P SQG K(QXL® 520)-NH2 and incubated for 5 minutes in a white 96-well ProxiPlate (Perkin Elmer). The specific FAP substrate used was the FRET-peptide HiLyteFluor™ 488-VS(D-)P SQG K(QXL® 520)-NH2 (Bainbridge, et al., Sci Rep, 2017, 7:12524). 25 μl of a 30 μM substrate solution diluted in assay buffer was added. All solutions were equilibrated at 37°C before use. Substrate cleavage and the increase in fluorescence (excitation at 485 nm and emission at 538 nm) were measured in kinetic mode over 5 minutes at 37°C using a SPECTRAmax M5 plate reader. RFU / sec were calculated using SoftMax Pro software and plotted against peptide concentration. Four-parameter logistic (4PL) curve fitting and pIC50 calculation were performed using ActivityBase software. The results of this assay for each compound of the invention are shown in Table 6 (Example 6). pIC50 Category A represents a pIC50 value of greater than 8.0, Category B represents a pIC50 value of 7.1-8.0, Category C represents a pIC50 value of 6.1-7.0, and Category D represents a pIC50 value of 6.0 or less.

[0325] As can be seen from Table 6, compounds of the present invention show surprisingly superior results in both the FACS binding assay and the FAP protease activity assay.

[0326] [Table 6-1]

[0327] [Table 6-2]

[0328] Example 8 Surface plasmon resonance assay Surface plasmon resonance studies were performed using a Biacore™ T200 SPR system. Briefly, polarized light is directed toward a gold-labeled sensor surface, and the minimum intensity of reflected light is detected. The angle of the reflected light changes as molecules bind and dissociate. The gold-labeled sensor surface is loaded with a FAP antibody bearing the FAP target protein, so that antibody binding does not occur at the FAP's substrate-binding site. A test compound is contacted with the loaded surface, and a real-time interaction profile with the FAP ligand is recorded in a sensorgram. The association and dissociation of the binding interaction are measured in real time, allowing the association and dissociation rate constants and corresponding affinity constants to be calculated. Importantly, background responses occur due to the refractive index difference between the running buffer and the sample buffer and nonspecific binding of the test compound to the flow cell surface. This background is measured and subtracted by running the sample over a control flow cell coated with the same density of capture antibody in the absence of immobilized FAP. Furthermore, baseline drift correction for the binding data, caused by the gradual dissociation of the captured FAP from the immobilized antibody, is performed. This drift is measured by injecting running buffer through a flow cell with antibody and FAP immobilized on the sensor surface.

[0329] A Biacore™ CM5 sensor chip was used. Human anti-FAP antibody (MAB3715, R&D Systems) was diluted to a final concentration of 50 μg / ml in 10 mM acetate buffer, pH 4.5. 150 μL aliquots were transferred to plastic vials and placed in the sample rack of a Biacore™ T200 instrument. The following Amine Coupling Kit Reagent solutions were transferred to plastic vials and placed in the sample rack: 90 μL of 0.4 M 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 90 μL of 0.1 M N-hydroxysuccinimide (NHS). A 130 μL aliquot of 1 M ethanolamine-HCl, pH 8.5, was transferred to a plastic vial and placed in the sample rack. The Biacore™ liquid system was set up as follows: distilled water (1 L), separate bottles containing running buffer (500 mL), and an empty bottle for waste were placed in the buffer tray. A pre-installed program for immobilization was used, and the immobilization level was 7000 RU. Immobilization was performed at 25°C. The immobilization procedure for anti-FAP antibodies was performed as described in Table 7.

[0330] [Table 7]

[0331] Human recombinant FAP was diluted in running buffer to a final concentration of 20 μg / mL. 100 μL aliquots of the human FAP working solution were transferred to plastic vials and placed in a sample rack. 0.5 mM compound stock solutions were prepared by dissolving each compound in DMSO. For each test compound, the compound stock solution was diluted at 500 nM in running buffer (HBST) and diluted with HBST-DMSO buffer (0.1% DMSO). Further dilutions were performed. SPR binding analysis of the binary complexes was performed in SCK mode at 25°C. Table 8 describes the protocol for capturing and evaluating binding kinetics. After three SCK measurements, baseline drift was assessed by injecting running buffer through the flow cell where the antibody and FAP were immobilized on the sensor surface.

[0332] [Table 8]

[0333] For each test compound, the raw SPR data in the form of resonance units (RU) were plotted as a sensorgram using the Biacore™ T200 control software. The signal from the blank sensorgram was subtracted from the signal of the test compound sensorgram (blank correction). The blank-corrected sensorgram was corrected for baseline drift by subtracting the sensorgram of an SCK run (running buffer only) without test compound. The association rate (k on ), dissociation rate (k off ), dissociation constant (K D ), and t 1 / 2 was calculated from the blank-normalized SPR data. The raw data and fit results were imported as text files in IDBS. pK D Values ​​(negative decimal logarithms of the dissociation constants) were calculated using the IDBS Excel template.

[0334] The results of this assay for a selection of compounds of the present invention are shown in Table 9. Category A is a compound with a pK greater than 8.0 D Category B represents a pK value of 7.1 to 8.0. D Category C represents a pK value of 6.1 to 7.0 D Represents a value.

[0335] [Table 9]

[0336] Example 9 PREP and DPP4 protease activity assays To test the selectivity of the FAP-binding peptides for both PREP and DPP4, a protease activity assay was performed similar to the FAP activity assay described above, with the following exceptions.

[0337] PREP activity was measured using recombinant human PREP (R&D systems, #4308-SE). 50 μM Z-GP-AMC (Bachem, #4002518) was used as a substrate. DPP4 activity assays were performed in DPP assay buffer (25 mM Tris, pH 8.0). Recombinant human DPP4 was purchased from R&D systems (#9168-SE). 20 μM GP-AMC (Santa Cruz) was used as a substrate. Biotechnology, #115035-46-6) was used.

[0338] The fluorescence of AMC after cleavage (excitation at 380 nm and emission at 460 nm) was measured in kinetic mode over 5 minutes at 37°C on a SPECTRAmax M5 plate reader. RFU / sec was calculated using SoftMax Pro software and plotted against peptide concentration. Four-parameter logistic (4PL) curve fitting and pIC50 calculation were performed using ActivityBase software. The results of this assay for some of the compounds of the present invention are shown in Table 10 below.

[0339] [Table 10]

[0340] Example 10 Specificity Screening To early identify significant off-target interactions of the compounds of the present invention, specificity screening was performed. Bowes, et al. (Nat Rev Drug 44 selected targets recommended by Discov, 2012, 11:909 Specificity was tested using a standard battery of assays ("SafetyScreen44™ Panel") containing a target and a compound that binds to this target (referred to as "Ref. Compound"). The Reference Compound served as a positive control for each assay, and therefore inhibition would be expected to be detected by this Reference Compound. However, compounds of the present invention were not expected to show inhibition in this assay. These binding and enzyme inhibition assays were performed by Eurofins Cerep SA (Celle l'Evescault, France).

[0341] 3BP-3407 and 3BP-3554 were tested at 10 μM. Compound binding was calculated as the % inhibition of binding of the radioactively labeled ligand specific for each target ("% inhibition of specific binding" (3BP-3407) or (3BP-3554), respectively). The enzyme inhibitory effect of the compounds was calculated as the % inhibition of control enzyme activity.

[0342] Results showing greater than 50% inhibition or stimulation are considered to represent a significant effect of the test compound. No such effect was observed at any of the receptors studied, listed in Table 11 below. A summary of the results of this assay is summarized in Table 11 below.

[0343] [Table 11-1]

[0344] [Table 11-2]

[0345] [Table 11-3]

[0346] [Table 11-4]

[0347] [Table 11-5]

[0348] [Table 11-6]

[0349] Additionally, to further determine the specificity of the compounds of the present invention, specificity screening for proteases was performed by BPS Biosciences (Turk, Nat Rev Drug Discov, 2006, 5:785; Overall, et al., Nat Rev Cancer, 2006, 6:227; Anderson, et al., Handb Exp Pharmacol, 2009, 189:85).

[0350] 3BP-3407 and 3BP-3554 were tested in duplicate at 1 μM and 10 μM. In the absence of the compound, the fluorescence intensity (Ft) of each data set was defined as 100% activity. In the absence of the enzyme, the background fluorescence intensity (Fb) of each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following formula: activity % = (F-Fb) / (Ft-Fb), where F = fluorescence intensity in the presence of compound. The percentage of inhibition was calculated according to the following formula: inhibition % = 100% - activity %. Results showing greater than 50% inhibition were considered to represent a significant effect of the test compound. The results of this assay are shown in Table 12 below.

[0351] [Table 12-1]

[0352] [Table 12-2]

[0353] [Table 12-3]

[0354] [Table 12-4]

[0355] Example 11 of selected compounds 111 In- and 177 Lu-labeled To serve as a diagnostic, therapeutic, or diagnostically active agent, the compound must be labeled with a radioactive isotope. To ensure high radiochemical yield and purity of the radiolabeled compound of the present invention, the labeling procedure must be carried out appropriately. This example demonstrates that the compound of the present invention is suitable for radiolabeling and can be labeled with high radiochemical yield and purity.

[0356] 111 30–100 MBq of InCl3 (in 0.02 M HCl) was mixed with 1 nmol of compound (200 μM stock solution in 0.1 M HEPES pH 7) per 30 MBq and buffer (1 M sodium acetate buffer pH 5 or 1 M sodium acetate / ascorbic acid buffer pH 5 containing 25 mg / ml methionine) at a final buffer concentration of 0.1–0.2 M. The mixture was heated to 80°C for 20–30 min. After cooling, DTPA and TWEEN-20 were added to final concentrations of 0.2 mM and 0.1%, respectively.

[0357] 1770.2-2.0 GBq of LnCl (in 0.04 M HCl) was mixed with 1 nmol of compound (200 μM stock solution in 0.1 M HEPES pH 7) per 45 MBq and buffer (1 M sodium acetate / ascorbic acid buffer pH 5 containing 25 mg / ml methionine) for a final buffer concentration of approximately 0.4 M. The mixture was heated to 90°C for 20 min.

[0358] The labeling efficiency was analyzed by thin-layer chromatography (TLC) and HPLC. For TLC analysis, 1-2 μl of the diluted labeling solution was applied to a strip of iTLC-SG chromatography paper (Agilent, 7.6 × 2.3 mm) and developed with citrate-dextrose solution (Sigma). The iTLC strip was then cut into three pieces, and the associated radioactivity was measured in a gamma counter. The radioactivity measured at the solvent front represents the free radionuclide and colloid, while the radioactivity at the front represents the radiolabeled compound. For HPLC, 5 μl of the diluted labeling solution was applied to a Poroshell SB-C18 column. Analysis was performed on a 2.7 μm column (Agilent). Eluent A: MeCN, Eluent B: HO, 0.1% TFA, gradient from 5% B to 70% B within 15 min, flow rate: 0.5 ml / min; detector: NaI (Raytest), DAD 230 nm. The peak eluting at the dead volume represents the free radionuclide, while the peak eluting at the peptide-specific retention time determined by the unlabeled sample represents the radiolabeled compound.

[0359] At the end of the synthesis, radionuclide incorporation was greater than 95% and radiochemical purity was greater than 90%. 111 Exemplary radiochemical purities for In-labeled compounds are shown in Table 13. 177 The Lu-labeled compounds maintained radiochemical purity of 90% or more up to 6 days after synthesis (Table 14). Radiochromatograms for selected compounds are shown in Figures 1-4. Figure 1 shows the radiochromatograms of Lu-labeled compounds in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed immediately upon completion of synthesis. 177FIG. 2 shows the radiochromatogram of Lu-3BP-3407 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed 6 days after completion of synthesis. 177 Figure 3 shows the radiochromatogram of Lu-3BP-3407 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed immediately upon completion of the synthesis. 177 FIG. 4 shows the radiochromatogram of Lu-3BP-3554 in formulation buffer containing 100 mg / mL ascorbate and 5 mg / mL L-methionine, analyzed 6 days after completion of synthesis. 177 The emission chromatogram of Lu-3BP-3554 is shown.

[0360] [Table 13]

[0361] [Table 14]

[0362] Example 12 Imaging and biodistribution studies Radioactively labeled compounds can be detected by imaging methods such as SPECT and PET. Furthermore, the data obtained by such techniques can be confirmed by direct measurement of the radioactivity contained in individual organs prepared from animals injected with the radioactively labeled compounds of the present invention. Therefore, the biodistribution of radioactively labeled compounds (measurement of radioactivity in individual organs) can be determined and analyzed. This example demonstrates that the compounds of the present invention exhibit biodistribution suitable for tumor imaging diagnosis and therapeutic treatment.

[0363] All animal experiments were performed in compliance with German animal protection laws. Male SCID beige mice (6-8 weeks old, Charles River, Sulzfeld, Germany) were inoculated with 5 × 10 6Each mouse was inoculated with HEK-FAP (embryonic human kidney 293 cells genetically engineered to express high levels of FAP). 3 When it reaches a size of over 30MBq 111 In-labeled compounds of the present invention (diluted to 100 μL with PBS) were administered intravenously via the tail vein. Images were obtained using a NanoSPECT / CT system (Mediso Medical Imaging Systems, Budapest, Hungary) using the following exemplary acquisition and reconstruction parameters (Table 15).

[0364] [Table 15]

[0365] Imaging data were saved as DICOM files and analyzed using VivoQuant™ software (Invicro, Boston, USA). Results are expressed as a percentage of injected dose per gram of tissue (%ID / g). For biodistribution studies, animals were sacrificed by cervical dislocation 24 or 48 hours after injection and then dissected. Various organs and tissues were harvested and weighed, and radioactivity was determined by gamma counting. Two animals were used per time point. Results are expressed as a percentage of injected dose per gram of tissue (%ID / g).

[0366] The results of imaging and biodistribution studies for selected compounds are shown in Figures 5-7. Example 13 Efficacy Study - HEK-FAP Radioactively labeled compounds can be used for therapeutic and diagnostic applications in a variety of diseases, particularly cancer. This example demonstrates that the compounds of the present invention have antitumor activity suitable for the therapeutic treatment of tumors.

[0367] All animal experiments were performed in compliance with German animal protection laws. Female Swiss nude mice (7-8 weeks old, Charles River Laboratories, France) were inoculated with 5 × 106 HEK-FAP cells were inoculated, and tumors grew to 160 ± 44 mm 3 Treatment was administered when a mean tumor volume of 100 mg / kg was reached. Mice were divided into four different groups of 10 animals per group: Group 1 - vehicle control, Group 2 - cold compound. nat Lu-3BP-3554, group 3-30MBq 177 Lu-3BP-3554 (low dose), and group 4-60MBq 177 Lu-FAP-3554 (high dose). Treatment was administered by intravenous injection into the tail vein at 4 mL / kg (100 μL / mouse) on day 0. Tumor volume and body weight were measured on day 0 (i.e., day 1 of administration of the radiotracer) and then three times weekly until the completion of the study.

[0368] 177 The distribution of the tracer in mice injected with Lu-labeled 3BP-3554 was determined by SPECT imaging in groups of 3 mice per dose. After CT, CT scans were performed for anatomical information. Imaging was performed at 3, 24, 48, and 120 hours after injection with a NanoSPECT / CT system (Mediso Medical Imaging Systems, Budapest, Hungary) using the following exemplary acquisition and reconstruction parameters (Table 16).

[0369] [Table 16]

[0370] Imaging data were saved as DICOM files and analyzed using VivoQuant™ software (Invicro, Boston, USA). Results are expressed as percentage of injected dose per gram of tissue (%ID / g).

[0371] Vehicle-treated mice and cold compound nat The tumors in Lu-3BP-3554-treated mice were 1338 ± 670 mm on day 14, respectively.3 and 1392±420mm 3 A mean tumor volume (MTV) of 30 or 60 MBq was achieved (Figure 9A). Statistically significant (p<0.01) antitumor activity was observed in mice in both treatment groups. Tumor growth inhibition (TGI) at day 14 was 30 or 60 MBq. 177 Mice treated with a single dose of Lu-3BP-3554 had 111% and 1113%, respectively, compared to the vehicle-treated group. 177 The MTV of all mice treated with Lu-3BP-3554 was 70 mm on day 14. 3 Tumors were monitored for regrowth on day 42 (representing the end of the study) and treated with 30 or 60 MBq 177 Three of 10 and nine of 10 mice treated with Lu-3BP-3554 were tumor-free (10 mm 3 (<100 mg / kg bw) suggesting a potential dose-response in this model. No treatment-related weight loss was observed throughout the study (Figure 9B). After a 3-5% weight loss was observed in all groups on day 2, the animals' weights increased over time.

[0372] Both 177 SPECT / CT imaging of three animals in the Lu-labeled treatment group demonstrated high tumor-to-background contrast at all time points examined (3 to 120 hours post-injection (pi)). High tumor retention was observed up to 120 hours. The organ with the highest non-targeted uptake was the kidney, at 30 or 60 MBq. 177 8.6 ± 0.6 and 8.0 ± 1.6 at 3 hours p.i. in Lu-3BP-3554-treated mice, respectively. These ratios increased over time, reaching peak values ​​of 30 or 60 MBq at 120 hours. 177 Mice treated with Lu-3BP-3554 achieved tumor-to-kidney ratios of 40±7.9 and 32±7.4, respectively. An exemplary panel of SPECT / CT images for the high-dose animal, Mouse 5, is shown in FIG. 10A, and an exemplary panel of SPECT / CT images for the low-dose animal, Mouse 1, is shown in FIG. 10B.

[0373] Example 14 Imaging studies - Sarcoma PDX models Sarcoma tumors have been reported to express FAP, and imaging was performed to evaluate 3BP-3554 uptake in four different sarcoma patient-derived xenograft (PDX) tumor models. The Sarc4183, Sarc4605, Sarc4809, and Sarc12616 PDX models were derived from patients with rhabdomyosarcoma, osteosarcoma, undifferentiated sarcoma, and undifferentiated pleomorphic sarcoma, respectively (Experimental Pharmacology & Oncology Berlin-Buch, Germany). Tumor fragments were implanted subcutaneously into the left flank of 8-week-old NMRI nu / nu mice (Janvier Labs, France). All animal experiments were performed in compliance with German animal protection laws. At 47 days (Sarc4183, Sarc4809) or 46 days (Sarc4605, Sarc12616) after transplantation, 2–3 mice per model were treated with 30 MBq of PBS. 111 Images were taken 3 hours after a single intravenous injection of In-3BP-3554. Imaging was performed as described in Example 12.

[0374] 111 Imaging results with In-3BP-3554 demonstrated high tumor uptake at 3 hours post-injection and high tumor-to-background contrast. Representative SPECT / CT images are shown in Figure 11A. Quantification of tumor uptake in two PDX-bearing mice (Sarc4605, Sarc12616) or three PDX-bearing mice (Sarc4183, Sarc4809) yielded %ID / g values ​​of 4.9 ± 1.7 (Sarc4183), 5.2 ± 0.8 (Sarc4605), 4.4 ± 0.7 (Sarc4809), and 6.1 ± 0.6 (Sarc12616), respectively, as shown in Figure 11B. These results demonstrate the efficacy of In-3BP-3554 in all four sarcoma models. 111Uptake of In-3BP-3554 has been demonstrated. The tumor-to-kidney ratios were 4.7±1.2 (Sarc4183), 3.2±0.4 (Sarc4605), 4.1±0.7 (Sarc4809), and 4.3±1.2 (Sarc12616).

[0375] Example 15 Efficacy Study - Sarcoma Sarc4809 PDX Model 177 The efficacy of Lu-3BP-3554 was investigated in the human sarcoma PDX tumor model Sarc4809, an undifferentiated sarcoma model. 111 It was shown to uptake In-3BP-3554 (Example 14) and also to express FAP by immunohistochemistry.

[0376] All animal experiments were performed in compliance with German animal protection laws. Sarc4809 tumor fragments were implanted subcutaneously into the left flank of 8-week-old NMRI nu / nu mice (Janvier Labs, France). 187.08 ± 123.8 mm 3 Treatment began 23 days after implantation with a mean tumor volume of 1.0001. The mice were divided into four groups of 10 animals per group: Group 1 - vehicle control, Group 2 - cold compound. nat Lu-3BP-3554, group 3-30MBq 177 Lu-3BP-3554, group 4-60MBq 177 Lu-FAP-3554. Treatment was administered by intravenous injection into the tail vein at 4 mL / kg (100 μL / mouse) on day 0. Tumor volume and body weight were determined on day 0 (i.e., the first day of radiotracer administration) and then three times a week until the completion of the study.

[0377] All tumors continued to grow throughout the follow-up period of this study up to day 42. Vehicle-treated mice and nat Tumors in Lu-3BP-3554-treated mice (control group) each grew to 894 ± 610 mm on day 31 (the last day that at least 50% of mice per group were still alive). 3 and 1225±775mm 3MTV reached 30 or 60MBq 177 Tumors in mice treated with a single dose of Lu-3BP-3554 grew to 635 ± 462 and 723 ± 391 mm at day 31, respectively. 3 The MTV of 30 or 60 MBq was reached (Figure 12A). Statistically significant (p<0.05) antitumor activity was observed in mice in both treatment groups. 177 In mice treated with a single dose of Lu-3BP-3554, tumor growth inhibition (TGI) at day 31 was 61% and 73%, respectively, compared to the vehicle-treated group. No treatment-related body weight loss (BWL) was observed during this study. All groups gained weight during the follow-up of this study (Figure 12B).

[0378] Example 16 Pharmacokinetic studies The pharmacokinetic behavior of selected compounds was evaluated in mice and rats. Characterization of the pharmacokinetic behavior of the compounds provides new insights into the distribution and excretion of the compounds, as well as exposure calculations.

[0379] Various amounts of compounds were stably formulated in PBS. These formulations were administered intravenously to mice at doses of 4 nmol / kg, 40 nmol / kg, and 400 nmol / kg, and to rats at doses of 2 nmol / kg, 20 nmol / kg, and 200 nmol / kg (3BP-3554) or 40 nmol / kg and 400 nmol / kg (3BP-3623). Assuming an allometric translation factor of 12.3 from humans to mice and 6.2 from humans to rats (Nair AB, Jacob S. Journal of Basic and Clinical Pharmacy, 2016, 7(2): 27-31), the applied doses represent a human dose range of 0.325 nmol / kg to 32.5 nmol / kg.

[0380] Blood samples were taken from the tail vein (rats) or retrobulbar (mice) after various times (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h). After separating blood cells from plasma by centrifugation, the prepared plasma samples were subjected to a protein precipitation procedure to quantify the compounds. 150 μl of zinc sulfate precipitant containing 78% 0.1 M zinc sulfate and 22% acetonitrile was added. After incubation at room temperature for 30 minutes, the precipitate was separated by centrifugation. If the compound contained a free DOTA moiety, 10 μl of 1% formic acid was added to 100 μl of the supernatant, followed by further incubation at 60°C for 10 minutes to complete the formation of the zinc chelate.

[0381] The determination of analytes in clean sample solutions was carried out on an Agilent 1290 UHPLC system coupled with an Agilent 6470 triple quadrupole mass spectrometer. Chromatographic separation was carried out by gradient elution using a mixture of 0.1% formic acid in water as eluent A and acetonitrile as eluent B (isocratic at 5% B over 1 min, followed by a linear gradient to 43% B in 4 min, 500 μl / min) on a Phenomenex column at 40°C. It was performed on a BioZen Peptide XB-C18 HPLC column (50 × 2 mm, 1.7 μm particle size).

[0382] Mass spectrometric detection was performed in positive ion ESI mode with multiple reaction monitoring (MRM).

[0383] [Table 17]

[0384] Quantitation of test items was achieved using the Quantitative Analysis software from the Agilent MassHunter software suite. Quadratic regression was performed with a weighting factor of 1 / x.

[0385] Plasma levels were subjected to non-compartmental analysis (NCA) and the following results were obtained: initial concentration of the compound (C0), volume of distribution at steady state (V ss ), terminal volume of distribution (V z ), terminal half-life (t 1 / 2 ), clearance (CL), and area under the curve extrapolated to infinity (AUC inf The NCA parameters of 3BP-3554 are summarized in Table 18 for 3BP-3554 in mouse plasma and in Table 19 for 3BP-3554 in rat plasma, and the NCA parameters of 3BP-3623 are summarized in Table 20 for 3BP-3623 in mouse plasma and in Table 21 for 3BP-3623 in rat plasma.

[0386] [Table 18]

[0387] [Table 19]

[0388] [Table 20]

[0389] [Table 21]

[0390] These results indicate distribution primarily in blood and interstitial fluid, with clearance typical of peptides, with terminal half-lives ranging from 23 to 59 minutes in mice and 45 to 71 minutes in rats. Exposure, as described by AUC, correlates approximately linearly with the injected dose, and clearance is constant for all applied doses in a particular animal model. These observations suggest that there is no significant nonlinearity in pharmacokinetic behavior that needs to be taken into account when calculating the initial dose in humans.

[0391] The features of the invention disclosed in the specification, the claims, the sequence listing and / or the drawings may, both separately and in any combination thereof, be material for realizing the invention in diverse forms thereof. References The disclosure of each and every document recited herein is incorporated by reference. Without being limited thereto, the present invention includes the following aspects. [Aspect 1] The following formula [ka] the compound Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554), and the compound of the formula [ka] The compound Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407) A compound selected from the group consisting of: [Aspect 2] The following formula [ka] 2. The compound according to embodiment 1, wherein the compound is Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554) of the formula [Aspect 3] The following formula [ka] 2. The compound according to embodiment 1, wherein the compound is Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3407). [Aspect 4] The compound comprises a diagnostically active nuclide or a therapeutically active nuclide, preferably the diagnostically active nuclide is a diagnostically active radionuclide, more preferably 43 Sc, 44 Sc, 51 Mn, 52 Mn, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 94m Tc, 99m Tc, 111 In, 152 Tb, 155 Tb, 201 Tl, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, preferably 43 Sc, 44 Sc, 64 Cu, 67 Ga, 68 Ga, 86 Y, 89 Zr, 99m Tc, 111 In, 152 Tb, 155 Tb, 203 Pb, 18 F, 76 Br, 77 Br, 123 I, 124 I, 125 I, most preferably 64 Cu, 68 Ga, 89 Zr, 99m Tc, 111 In, 18 F, 123 I, and 124 I, wherein said therapeutically active nuclide is a therapeutically active radionuclide, more preferably 47 Sc, 67 Cu, 89 Sr, 90 Y, 153 Sm, 149 Tb, 161 Tb,177 Lu, 186 Re, 188 Re, 212 Pb, 213 Bi, 223 Ra, 225 Ac, 226 Th, 227 Th, 131 I, 211 At, preferably 47 Sc, 67 Cu, 90 Y, 177 Lu, 188 Re, 212 Pb, 213 Bi, 225 Ac, 227 Th, 131 I, 211 At, most preferably 90 Y, 177 Lu, 225 Ac, 227 Th, 131 I, and 211 A compound according to any one of aspects 1 to 3, selected from the group consisting of At. [Aspect 5] The following formula [ka] The compound Hex-[Cys(tMeBn(InDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3590), The following formula [ka] The compound Hex-[Cys(tMeBn(LuDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3591), The following formula [ka] The compound Hex-[Cys(tMeBn(GaDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3592), The following formula [ka] The compound Hex-[Cys(tMeBn(EuDOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2 (3BP-3661), The following formula [ka] The compound Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3623), The following formula [ka] The compound Hex-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3624), The following formula [ka] The compound Hex-[Cys(tMeBn(EuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3662), The following formula [ka] The compound Hex-[Cys(tMeBn(GaDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3949), The following formula [ka] the compound Hex-[Cys-(tMeBn(CuDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4293), and The following formula [ka] The compound Hex-[Cys-(tMeBn(ZnDOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4343) 5. The compound according to embodiment 4, selected from the group comprising: [Aspect 6] 5. A compound according to any one of aspects 1 to 5 for use in a method for the diagnosis of a disease, for use in a method for the treatment of a disease, for use in a method for identifying a subject, wherein said subject is likely to respond or not likely to respond to treatment of a disease, said method for identifying a subject comprising the step of performing a method of diagnosis using a compound according to any one of aspects 1 to 5, preferably a method for the diagnosis of a disease as described in any one of aspects 1 to 5, or for use in a method for selecting subjects from a group of subjects, wherein said subjects are likely to respond or not likely to respond to treatment of a disease, said method for selecting subjects from a group of subjects comprising the step of performing a method of diagnosis using a compound according to any one of aspects 1 to 5, preferably a method for the diagnosis of a disease as described in any one of aspects 1 to 5, or a method for stratifying a group of subjects into subjects likely to respond to treatment of a disease and subjects not likely to respond to treatment of a disease, wherein said method for stratifying a group of subjects comprises the step of performing a method of diagnosis using a compound according to any one of aspects 1 to 5, preferably a method for the diagnosis of a disease as described in any one of aspects 1 to 5. [Aspect 7] A composition, preferably a pharmaceutical composition, comprising a compound according to any one of aspects 1 to 5 and a pharmaceutically acceptable excipient. [Aspect 8] 6. A kit comprising a compound of any one of aspects 1 to 5, optionally one or more excipients, and optionally one or more devices, said devices being selected from the group comprising a labeling device, a purification device, a handling device, a radiation protection device, an analytical device, or an administration device.

Claims

[Claim 1] The invention as described in the specification and drawings.

Images