COMPOUNDS COMPRISING A FIBROBLAST ACTIVATION PROTEIN LIGAND AND USE THEREOF

FIELD OF INVENTION

The present invention is related to a chemical compound; an inhibitor of fibroblast activation protein (FAP); a composition comprising the compound and inhibitor, respectively; the compound, the inhibitor and the composition, respectively, for use in a method for the diagnosis of a disease; the compound, the inhibitor and the composition, respectively, for use in a method for the treatment of a disease; the compound, the inhibitor and the composition, respectively, for use in a method of diagnosis and treatment of a disease which is also referred to as “thera(g)nosis” or “thera(g)nostics”; the compound, the inhibitor and the composition, respectively, for use in a method for delivering an effector to a FAP-expressing tissue; a method for the diagnosis of a disease using the compound, the inhibitor and the composition, respectively; a method for the treatment of a disease using the compound, the inhibitor and the composition, respectively; a method for the diagnosis and treatment of a disease which is also referred to as “thera(g)nosis” or “thera(g)nostics”, using the compound, the inhibitor and the composition, respectively; a method for the delivery of an effector to a FAP-expressing tissue using the compound, the inhibitor and the composition, respectively.

BACKGROUND

Despite the increasing availability of therapeutic options, cancer is still the second leading cause of death globally. Therapeutic strategies mainly focus on targeting malignant cancer cells itself, ignoring the ever-present surrounding tumor microenvironment (TME) that limit 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 not only of the heterogeneous population of cancer cells but also of a variety of resident and infiltrating host cells, secreted factors, and extracellular matrix proteins (Quail, et al., Nat Med, 2013, 19: 1423). A dominant cell type found in the TME is the cancer associated fibroblast (CAF) (Kalluri, Nat Rev Cancer, 2016, 16: 582). Many different cell types have been described as the source and origin for CAFs, such as e.g. 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 features and often are the dominant cell type within a solid tumor mass. CAFs have attracted increasing attention as a player in tumor progression and homeostasis (Gascard, et al., Genes Dev, 2016, 30: 1002; LeBleu, et al., Dis Model Mech, 2018, 11).

During recent years, fibroblast activation protein (FAP) has gained notoriety 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 omnipresence of CAFs and stroma within tumors, FAP was discovered as a suitable marker for radiopharmaceutical diagnostics and as a suitable target for radiopharmaceutical therapy (Siveke, J Nucl Med, 2018, 59: 1412).

Fibroblast activation protein α (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, DPP9), FAP has post-proline exopeptidase activity. In addition, FAP possesses endopeptidase activity, similar to prolyl oligopeptidase/endopeptidase (POP/PREP). The FAP gene is highly conserved across various species. The extracellular domain of human FAP shares 90% amino acid sequence identity with mouse and rat FAP. Mouse FAP has 97% sequence identity with rat FAP.

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 and an α/βhydrolase domain. The catalytic triad is composed of Ser624, Asp702, and His734 and is located at the interface of the β-propeller and the hydrolase domain. The active site is accessible through a central hole of the β-propeller domain or through a narrow cavity between the β-propeller and the hydrolase domain. FAP monomers are not active, but form active homodimers as well as heterodimers with DPP4 (Ghersi, et al., Cancer Res, 2006, 66: 4652). Soluble homodimeric FAP has also been described (Keane, et al., FEBS Open Bio, 2013, 4: 43; Lee, et al., Blood, 2006, 107:1397).

FAP possesses dual enzyme activity (Hamson, et al., Proteomics Cin Appl, 2014, 8: 454). Its dipeptidyl peptidase activity allows cleaving two amino acids of the N-terminus after a proline residue. FAP substrates that are cleaved rapidly via its dipeptidyl peptidase activity are neuropeptide Y, Peptide YY, Substance P, and B-type natriuretic peptide. Collagen I and III, Fibrobast Growth Factor 21 (FGF21) and α₂-antiplasmin have been shown to be cleaved by the endopeptidase activity of FAP. While FAP is unable to cleave native collagens, pre-digestion by other proteases, such as matrix metalloproteinases, facilitates further collagen cleavage by FAP. Processing of collagen may influence migratory capacities of cancer cells. Besides increasing invasiveness of cancer cells through remodeling of the extracellular matrix, several other FAP-mediated tumor promoting roles have been proposed, including proliferation and increasing angiogenesis. Furthermore, stromal expression of FAP is linked to escape from immunosurveillance in various cancers, suggesting a role in anti-tumor immunity (Pure, et al., Oncogene, 2018, 37: 4343).

FAP is transiently expressed during normal development, but only rarely in healthy adult tissues. In transgenic mice, it was demonstrated that FAP is 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, a FAP knockout mouse has 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).

FAP expression in the tumor stroma of 90% of epithelial carcinomas was first reported in 1990 under use of a 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 (CAF) and cancer-associated pericytes (Cremasco, et al., Cancer Immunol Res, 2018, 6: 1472). FAP expression on malignant epithelial cells has also been reported but its significance remains to be defined (Pure, et al., Oncogene, 2018, 37: 4343). The following Table 1, taken from Busek et al. (Busek, et al., Front Biosci (Landmark Ed), 2018, 23: 1933), summarizes the expression of FAP in various malignancies indicating the tumor type and the cellular expression.

TABLE 1

FAP expression in human malignancies (from Busek et al.)

Expression
Expression

of FAP in
of FAP in

Malignant
Stroma

Tumor Type
Cells
Cells
Notes

Basal cell carcinoma,
−
+
Expression in fibroblasts strongest in close proximity to cancer cells. FAP expression is

squamous cell

absent in benign epithelial tumors, its positivity in the stroma may be a useful criterion

carcinoma of the skin

for differentiating between morpheaform/infiltrative basal cell carcinomas and FAP-

negative desmoplastic trichoepithelioma.

Oral squamous cell
+
+
FAP is a negative prognostic marker - elevated expression is associated with greater

carcinoma

tumor size, lymph-node metastasis, advanced clinical stage, and worse overall survival.

Melanoma
−
+
FAP expression present in a subset of melanocytes in 30% of benign melanocytic nevi,

(in situ)

but not detectable in malignant melanoma cells in melanoma tissues. The quantity of

FAP-positive stromal cells is positively associated with ECM content and inflammatory

cell infiltration. Normal melanocytes express FAP in vitro. Conflicting data for FAP in

melanoma cells: several human melanoma cell lines express FAP and FAP contributes to

their invasiveness in vitro, but immunopositivity has not been detected in melanoma

tissues. Mouse melanoma cell lines are FAP-negative and mouse FAP is a tumor

suppressor independently of its enzymatic activity.

Esophageal cancer
+
+
FAP is expressed in cancer cells as well as in premalignant metaplastic cells of the

esophagus in both adenocarcinoma and squamous cell carcinoma.

Gastric cancer
+
+
A higher stromal FAP expression at the invasion front is associated with low tumor cell

(incl. low
differentiation, more advanced TNM stage, serosal invasion, and poor survival. A higher

expression
stromal FAP is associated with worse survival. A higher FAP expression in intestinal-type

in endo-
gastric cancer (in stroma, moderately differentiated cancer cells, and endothelial cells)

thelial cells)
than in the diffuse type (mainly in cancer cells with poor cell-to-cell contacts, endothelial

cells). A higher stromal FAP expression in the intestinal-type gastric cancer is associated

with the presence of liver and lymph node metastases.

Colorectal cancer
+
+
A higher stromal FAP positivity found in earlier-stage disease, but in patients with stage

IV tumors high FAP is associated with worse survival. A higher FAP expression is

associated with advanced Duke stage. A high FAP expression in the tumor center is a

negative prognostic factor. Stromal FAP expression in stage II/III rectal cancer after

chemoradiotherapy is associated with a worse prognosis. A higher FAP mRNA expression

is associated with worse disease-free survival and a trend for worse overall survival.

Pancreatic
+
+
FAP expression in carcinoma cells is associated with a larger tumor size, presence of a

adenocarcinoma

fibrotic focus, perineural invasion, and a worse prognosis. Stromal FAP expression

correlates with lymph node metastasis and reduced survival. Nevertheless, a recent

retrospective Korean study reports an association between a lower number of FAP+

fibroblasts and a decreased overall survival based on a univariate analysis.

Hepatocellular carcinoma

+
FAP expression detected especially in tumors with abundant fibrous stroma. FAP mRNA

expression increased in peritumoral tissue, positively correlating with the density of

peritumoral activated HSCs. Higher levels are associated with more frequent early

recurrence, larger tumor size, presence of vascular invasion, and an advanced TNM stage.

Non-small cell lung
−/+
+
Absence of stromal FAP expression (24% of cases) in NSCLC is associated with better

cancer

survival. Reports regarding expression in cancer cells are inconsistent.

Mesothelioma
+
+
Expression, although to a variable extent, has been detected in all subtypes.

Breast tumors
+
+
FAP positivity detected mainly in the stroma; another study proposes a predominant

(ductal
(incl. endo-
localization in cancer cells in ductal adenocarcinoma. Jung et al. observed expression in

adenocarci-
thelial cells
cancer and stromal cells in 50% of cases where stroma is rich in adipose tissue

noma)

(approximately 1/3 of all tumors); in these cases, FAP expression was associated with a

higher tumor grade. In tumors with fibrous stroma, FAP expression was virtually absent

(2/3 of all tumors)

FAP expression is higher in cancer cells in lobular cancer than in ductal carcinoma. Stromal

FAP and calponin positivity may be an ancillary marker for detecting microinvasion in

ductal carcinoma. FAP expression increases with the malignant progression of phyllodes

tumors, but a later study detected stromal FAP expression only in 12.5% of the malignant

phyllodes tumors by IHC. Conflicting data regarding a possible association with breast

cancer survival: smaller studies have reported that a higher total FAP mRNA expression

is associated with worse survival, while a higher stromal FAP expression detected by IHC

was associated with a longer overall survival and disease-free survival. A recent larger

study involving 939 breast cancer patients did not prove any association between FAP

expression in the cancer or stromal cells and survival.

Renal cancer
−
+
Stromal FAP expression (detected in 23% of cases) associated with markers of

aggressiveness and worse survival in clear cell renal cell carcinoma. In metastatic clear

cell renal carcinoma, stromal FAP expression was detected in 36% of primary and 44% of

metastatic lesions, and was associated with several parameters of tumor aggressiveness

and worse survival.

Prostate cancer
−
+
Only small patient cohorts reported in literature. Expression in stromal cells detected in

7/7 cases, most intense in stromal cells adjacent to cancer cells.

Cervical cancer
+
+
No FAP expression was detected in preinvasive cervical neoplasia (CIN1, 2), occasional

positivity in stroma in CIN3 with moderate or severe inflammatory infiltrates. Enhanced

expression of FAP was found in cancer cells and subepithelial stromal cells in some of the

microinvasive and all of the invasive carcinomas.

Ovary
+
+
FAP positivity increases with tumor stage; negative FAP expression is associated with

longer disease-free survival. FAP positivity detected in cancer cells in 21% of tumors,

stromal positivity in 61%. Another study reported stromal positivity in 92% of cancer

tissues with extremely rare FAP expression in malignant cells; it also reported an

association with advanced tumor stage and presence of lymph node metastases. FAP-

positive malignant cells are present in malignant pleural and peritoneal effusions: strong

positivity is associated with worse survival.

Glioma
+
+
FAP expression increased in glioblastoma, highest expression found in the mesenchymal

subtype and gliosarcoma. Low expression in glioma stem-like cells. In glioblastoma,

overall FAP quantity is not associated with survival.

Thyroid cancer
−
+
FAP upregulated in aggressive papillary thyroid carcinomas. In medullary thyroid

carcinoma, FAP expression in the peritumoral and intratumoral stromal compartment

correlates with the degree of desmoplasia and presence of lymph node metastases.

Parathyroid tumors
n.d.
+
FAP mRNA expression was significantly higher in parathyroid carcinomas than in

adenomas.

Sarcomas
+
+
FAP expression found in malignant cells in fibrosarcomas, leiomyosarcoma, malignant

(see note)
(reactive
fibrous histiocytoma, low grade myofibroblastic sarcoma, fibroblastic areas in

fibroblasts
osteosarcomas, osteoid osteoma, and in osteosarcoma. FAP is negative in malignant cells

in Ewing's
with ″small round cell″ phenotype (embryonal rhabdomyosarcoma, Ewing sarcoma, or

sarcomas)
mesenchymal chondrosarcoma). A higher expression in osteosarcoma associated with

more advanced clinical stage, presence of distant metastasis, high histological grade, and

a worse progression-free and overall survival. FAP is expressed in both malignant and

benign tumors and its positivity reflects their histogenetic origin rather than malignant

potential.

Myeloma
−
+
FAP expression was detected in osteoclasts, endothelial cells, adipocytes, fibrotic stroma,

but not in multiple myeloma cells. FAP is upregulated in osteoclasts co-cultured with

myeloma cells.

FAP expression in CAFs was shown for almost 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). Utilization of FAP as a therapeutic target is therefore not limited to certain tumor entities.

The abundance of FAP-expressing CAFs is described to correlate with poor prognosis. Across a wide range of human tumor indications, FAP expression is described to correlate with higher tumor grade and worse overall survival (Pure, et al., Oncogene, 2018, 37: 4343).

As described above, it is indicated that FAP as well as FAP-expressing cells present in the tumor microenvironment significantly influence tumor progression (Hanahan, et al., Cancer Cell, 2012, 21: 309). Additionally, due to its relatively selective expression in tumors, FAP is regarded as 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).

Soon after its discovery, FAP was utilized as a therapeutic target in cancer. Until today, various strategies have been explored, including e.g. inhibition of FAP enzymatic activity, ablation of FAP-positive cells, or targeted delivery of cytotoxic compounds.

In 2007, an inhibitor of FAP and DPP4, Talabostat (Val-boro-Pro, PT-100), was developed by Point Therapeutics (for example as described in U.S. Pat. No. 6,890,904 or published international patent application WO9916864). Pennisi et al. (Pennisi, et al., Br J Haematol, 2009, 145: 775) observed a reduced tumor growth in a multiple myeloma animal model as well as in cancer syngeneic mouse models. Furthermore, several other prolyl boronic acid derivatives have been developed and reported as putative selective inhibitors for FAP. These derivatives show instability in aqueous environments at physiologic pH (Coutts, et al., J Med Chem, 1996, 39: 2087) and a non-specific reactivity with other enzymes.

WO 2008/116054 disclosed hexapeptide derivatives wherein compounds comprise a C-terminal bis-amino or boronic acid functional group.

US 2017/0066800 disclosed pseudopeptide inhibitors, such as M83, effective against FAP. These inhibitors were assessed in lung and colon cancer xenografts in immunodeficient mice. A suppression of tumor growth was observed (Jackson, et al., Neoplasia, 2015, 17: 43). These pseudopeptides inhibit the activity of both prolyl oligopeptidase (POP/PREP) and FAP, thereby excluding their use as specific therapeutic FAP inhibitors.

US 2008/280856 disclosed a nanomolar boronic acid-based inhibitor. The inhibitor shows a bispecific inhibition of FAP and PREP, thereby excluding their use as specific therapeutic FAP inhibitors.

FAP inhibitors based on cyclic peptides were disclosed, e.g., in WO 2016/146174 and WO 2006/042282. WO 2016/146174 disclosed peptides for diagnosis and treatment of tumors expressing FAP showing specificity for FAP, whereby closely related homologue DPP4 was not recognized by said peptides. WO 2006/042282 disclosed polypeptides for treatment of melanoma. In nude mice, inhibition of melanoma growth and melanoma metastasis was shown.

WO 99/75151 and WO 01/68708 disclosed a humanized FAP monoclonal antibody, F19, (Sibrotuzumab). Furthermore, the anti-FAP antibody F19 and humanized versions thereof were disclosed in WO 99/57151 and WO 01/68708. Development approaches involved, e.g., the generation of high affinity, species cross-reactive, FAP-specific scFvs converted into a bivalent derivative (Brocks, et al., Mol Med, 2001, 7: 461). In Phase I and II clinical trials, Sibrotuzumab showed specific tumor enrichment whilst failing to demonstrate measurable therapeutic activity in patients with metastatic colorectal cancer, with only 2 out of 17 patients having stable disease (Hofheinz, et al., Onkologie, 2003, 26: 44). This F19 antibody has not been shown to block any cellular or protease function of FAP, which might explain the lack of therapeutic effects (Hofheinz, et al., Onkologie, 2003, 26: 44; Scott, et al., Clin Cancer Res, 2003, 9: 1639).

US 2018/022822 disclosed novel molecules specifically binding to human FAP and epitopes thereof, 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 syngeneic MC38 colorectal tumors with an anti-FAP antibody reduced the tumor diameter and number of metastasis. WO 2012/020006 disclosed glycoengineered antibodies that bear modified oligosaccharides in the Fc region. Subsequently, bispecific antibodies specific for FAP and DR5 were developed as subject to WO 2014/161845. These antibodies trigger tumor cell apoptosis in vitro and in in vivo preclinical tumor models with FAP-positive stroma (Brunker, et al., Mol Cancer Ther, 2016, 15: 946). Antibody drug conjugates and immunotoxins that target FAP are described in WO 2015/118030. In vitro toxicity as well as in vivo inhibition of tumor growth was shown following application of anti-hu/moFAP hu36:cytolysin ADC candidates. It is unclear whether these antibodies were capable of inhibiting FAP activity.

Small molecule FAP inhibitors based on (4-quinolinoyl)glycyl-2-cyanopyrrolidine displaying low nanomolar inhibitory potency and high selectivity against related DPPs and PREP were 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 WO 2013/107820. However, the compounds are structurally unrelated to the compounds of the present invention and include a war-head leading to covalent binding to FAP.

In recent years, several FAP-targeted radiopharmaceutical approaches were developed which are exemplarily described herein.

WO 2010/036814 disclosed small molecule inhibitors of FAP for use as therapeutic agents through inhibition of FAPs enzyme activity or as radiopharmaceuticals through binding to FAP.

WO 2019/083990 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 described selective uptake in tumors of cancer patients of imaging and radiotherapeutic agents (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, March 8 (epub ahead of print); Giesel, et al., Eur J Nucl Med Mol Imaging, 2019, 46: 1754; Kratochwil, et al., J Nucl Med, 2019, 60: 801) based on 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).

Clinical assessments of a ¹³¹I-labeled, humanized form of the F19 antibody (sibrotuzumab) revealed a selective uptake by tumors but not by normal tissues in patients with colorectal carcinoma 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 antibodies that makes them unsuitable for a diagnostic, therapeutic, or theragnostic approach involving radionuclides.

WO 2011/040972 disclosed high-affinity antibodies recognizing both human and murine FAP antigen as potent radioimmunoconjugates. ESC11 IgG1 induces down modulation and internalization of surface FAP (Fischer, et al., Clin Cancer Res, 2012, 18: 6208). WO 2017/211809 disclosed tissue targeting thorium-227 complexes wherein the targeting moiety has specificity for FAP. However, the long circulation time of antibodies makes them unsuitable for a diagnostic, therapeutic, or theragnostic approach involving radionuclides.

FAP has also been described as being involved in other diseases than oncology indications, examples of which are given below.

Fibroblast-like synoviocytes in rheumatoid arthritic joints of patients show a 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 play an important role in organizing the structure of synovial tissue of joints by producing extracellular matrix components, recruiting infiltrating immune cells and secreting inflammatory mediators. Considerable evidence exists supporting a role for 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 has a pathological role in cartilage turnover at least by promotion of proteoglycan loss and subsequently cartilage degradation (Bauer, et al., Arthritis Res Ther, 2006, 8: R171; Waldele, et al., Arthritis Res Ther, 2015, 17: 12). Therefore, it might serve as a marker for patient stratification, for evaluation and follow-up of treatment success, or as a therapeutic target (Bauer, et al., Arthritis Res Ther, 2006, 8: R171). In mice, a treatment response was demonstrated using SPECT/CT imaging of a ^99mTc-labeled anti-FAP antibody (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).

Additionally, FAP was 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 healing process of wounds (Ramirez-Montagut, et al., Oncogene, 2004, 23: 5435). Jing et al. demonstrated a time-dependent course of change in FAP expression following burn wounds in rats (Jing, et al., Nan Fang Yi Ke Da Xue Xue Bao, 2013, 33: 615). Inhibiting of FAP activity in reactive wound fibroblasts in Keloid scars, common benign fibroproliferative reticular dermal lesions, might offer therapeutic option to prevent disease progression (Dienus, et al., Arch Dermatol Res, 2010, 302: 725).

In fibrotic diseases, upregulated expression of FAP was observed e.g. in idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis. In an ex vivo model for Crohn's disease, a chronic bowel inflammatory disease characterized by an excessive, misbalanced extracellular matrix (ECM) deposition, upregulated FAP expression was observed. FAP inhibition reconstituted extracellular matrix homeostasis (Truffi, et al., Inflamm Bowel Dis, 2018, 24: 332). Similar observations were made by Egger et al. (Egger, et al., Eur J Pharmacol, 2017, 809: 64) under use of a murine model of pulmonary fibrosis. Inhibition of FAP leads to reduced fibrotic pathology. FAP is also expressed in the tissue remodelling region in chronically injured liver (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).

FAP is expressed in arteriosclerotic lesions and upregulated in activated vascular smooth muscle cells (Monslow, et al., Circulation, 2013, 128: A17597). Monslow et al. showed that targeted inhibition of FAP in arteriosclerotic lesions may decrease overall lesion burden, inhibit inflammatory cell homing, and increase lesion stability through its ability to alter lesion architecture by favoring matrix-rich lesions over inflammation. More importantly, most of the arteriosclerotic pathologies share a common pathogenic feature: the rupture of an atherosclerotic plaque inducing arteriosclerotic 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 a critical trigger of acute coronary syndromes that may lead to myocardial infarction and sudden cardiac death. One of the key events in promoting plaque instability is the degradation of the fibrous cap, which exposes the underlying thrombogenic plaque core to the bloodstream, thereby causing thrombosis and subsequent vessel 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 fibrous caps (Brokopp, et al., Eur Heart J, 2011, 32: 2713). A radiolabeled tracer was developed and its applicability for atherosclerosis imaging shown (Meletta, et al., Molecules, 2015, 20: 2081).

DETAILED DESCRIPTION OF THE INVENTION

The problem underlying the present invention is the provision of a compound which is suitable as a diagnostic agent and/or a pharmaceutical agent, particularly if conjugated to a diagnostically and/or therapeutically active effector. A further problem underlying the present invention is the provision of a compound which is suitable as a diagnostic agent and/or a pharmaceutical agent, particularly if conjugated to a diagnostically and/or therapeutically active effector, whereby the compound is a potent inhibitor of FAP activity; preferably the pIC50 of the compound is equal to or greater than 6.0. A further problem underlying the present invention is the provision of a compound which is suitable as a diagnostic agent and/or a pharmaceutical agent, particularly if conjugated to a diagnostically and/or therapeutically active effector, in the diagnosis and/or therapy of a disease where the diseased cells and/or diseased tissues express FAP. A still further problem underlying the instant invention is the provision of a compound which is suitable for delivering a diagnostically and/or therapeutically effective agent to a diseased cell and/or diseased tissue, respectively, and more particularly a FAP-expressing diseased cell and/or diseased tissue, preferably the diseased tissue comprises or contains cancer associated fibroblasts. Also, a problem underlying the present invention is the provision of a method for the diagnosis of a disease, of a method for the treatment and/or prevention of a disease, and a method for the combined diagnosis and treatment of a disease; preferably such disease is a disease involving FAP-expressing cells and/or tissues, more particularly a FAP-expressing diseased cell and/or diseased tissue, preferably the diseased tissue comprises or contains cancer associated fibroblasts. A still further problem underlying the present invention is the provision of a method for the identification of a subject, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, a method for the selection of a subject from a group of subjects, wherein the subject is likely to respond or likely not to respond to a treatment of a disease. Also, a problem underlying the present invention is the provision of a pharmaceutical composition containing a compound having the characteristics as outlined above. Furthermore, a problem underlying the present invention is the provision of a kit which is suitable for use in any of the above methods.

There is a need for compounds that are suitable as a diagnostic agent and/or pharmaceutical agent, particularly if conjugated to a diagnostically and/or therapeutically active effector. Furthermore, there is a need for compounds that are suitable as a diagnostic agent and/or a pharmaceutical agent, particularly if conjugated to a diagnostically and/or therapeutically active effector, whereby the compound is a potent inhibitor of FAP activity; preferably the pIC50 of the compound is equal to or greater than 6.0. Further, there is a need for compounds suitable as diagnostic agents and/or pharmaceutical agents, particularly if conjugated to a diagnostically and/or therapeutically active effector, in the diagnosis and/or therapy of a disease where the diseased cells and/or diseased tissues express FAP. Furthermore, there is a need for a compound which is suitable for delivering a diagnostically and/or therapeutically effective agent to a diseased cell and/or diseased tissue, respectively, and more particularly a FAP-expressing diseased cell and/or diseased tissue, preferably the diseased tissue comprises or contains cancer associated fibroblasts. Also, there is a need for a method for the diagnosis of a disease, of a method for the treatment and/or prevention of a disease, and a method for the combined diagnosis and treatment of a disease; preferably such disease is a disease involving FAP-expressing cells and/or tissues, more particularly a FAP-expressing diseased cell and/or diseased tissue, preferably the diseased tissue comprises or contains cancer associated fibroblasts. Furthermore, there is a need for a method for the identification of a subject, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, a method for the selection of a subject from a group of subjects, wherein the subject is likely to respond or likely not to respond to a treatment of a disease. Further, there is a need for a pharmaceutical composition containing a compound having the characteristics as outlined above. Furthermore, there is a need for a kit which is suitable for use in any of the above methods. The present invention satisfies these needs.

These and other problems are solved by the subject matter of the attached claims.

These and other problems underlying the present invention are also solved by the following embodiments.

Embodiment 1. A compound comprising a cyclic peptide

of formula (I)

embedded image

and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

embedded image

- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

embedded image

- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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- wherein
- R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein R^7band R^7care each and independently (C₁-C₄)alkyl and
- t is 1 or 2;
- Yc is a structure of formula (X)

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- linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein
- the substitution pattern of the aromatic group in formula (X) is ortho, meta or para,
- n=0 or 1,
- t=1 or 2,
- Y¹is C—H or N,
- Y²is N or C—R^c1,
- R^c1is H or CH₂—R^c2and
- R^c2is a structure of formula (XI), (XII) or (XXII)

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wherein
- R^c3and R^c4are each and independently selected from the group consisting of H and (C₁-C₄)alkyl and
- u=1, 2, 3, 4, 5 or 6,
- x and y are each and independently 1, 2 or 3, and
- X═O or S

wherein in formulae (XI) and (XXII) one of the nitrogen atoms is attached to —CH₂— of R^c1and in formula (XII) —X— is attached to —CH₂— of R^c1; and

wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is R^a1—NH—C(O)—; wherein R^a1is selected from the group consisting of C₃alkyl, C₄alkyl or C₅alkyl, each and independently optionally substituted by up to two substituents each and independently selected from the group consisting of OH, F, COOH, (C₃-C₈)cycloalkyl, aryl, heteroaryl and (C₃-C₈)heterocycle, and wherein in (C₁-C₈)alkyl one of the —CH₂-groups is optionally replaced by —S— or —O—.

Embodiment 2. The compound of Embodiment 1, wherein R^a1is selected from the group consisting of C₃alkyl, C₄alkyl or C₅alkyl.

Embodiment 3. The compound of any one of Embodiments 1 and 2, wherein R^a1is C4 alkyl.

Embodiment 4. The compound of Embodiment 3, wherein R^a1is n-butyl.

Embodiment 5. The compound of any one of Embodiments 1 to 4, wherein Xaa1 is a D-amino acid residue selected from the group consisting of cys, hcy and pen, or Xaa1 is an L-amino acid residue selected from the group consisting of Cys, Hcy and Pen.

Embodiment 6. The compound of Embodiment 5, wherein Xaa1 is Cys.

Embodiment 7. The compound of any one of Embodiments 1, 2, 3, 4, 5 and 6, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro, Gly, Nmg and their derivatives.

Embodiment 8. The compound of Embodiment 7, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro and Nmg.

Embodiment 9. The compound of any one of Embodiments 7 and 8, wherein Xaa2 is an amino acid residue of Pro.

Embodiment 10. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8 and 9, wherein Xaa3 is an amino acid residue selected from the group consisting of Pro, Hyp, Tfp, Cfp, Dmp, Aze and Pip, and their derivatives.

Embodiment 11. The compound of Embodiment 10, wherein Xaa3 is an amino acid residue of Pro.

Embodiment 12. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11, wherein Xaa4 is an amino acid residue selected from the group consisting of Thr, Hse, Asn, Gln and Ser, and their derivatives.

Embodiment 13. The compound of Embodiment 12, wherein Xaa4 is Thr.

Embodiment 14. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 13, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu, and their derivatives.

Embodiment 15. The compound of Embodiment 14, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu.

Embodiment 16. The compound of Embodiment 15, wherein Xaa5 is an amino acid residue of Gln.

Embodiment 17. The compound of Embodiment 15, wherein Xaa5 is an amino acid residue of Glu.

Embodiment 18. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 and 17, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each and independently selected from the group consisting of H, methyl, ethyl, propyl and isopropyl,
- R^6crepresents from 0 to 3 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand C₁-C₄alkyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and

s is 0 or 1.

Embodiment 19. The compound of Embodiment 18, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each H
- R^6crepresents from 0 to 2 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand methyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and

s is 0.

Embodiment 20. The compound of any one of Embodiments 18 to 19, wherein Xaa6 is an amino acid residue selected from the group consisting of Phe, Ocf, Ppa, Thi, 1Ni, Otf, and Mpa, and their derivatives.

Embodiment 21. The compound of Embodiment 20, wherein Xaa6 is an amino acid residue of Phe.

Embodiment 22. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 and 21, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂, Cysol, AET, Hcy, cys, cys-OH, cys-NH₂and hcy.

Embodiment 23. The compound of Embodiment 22, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂, Cysol and AET.

Embodiment 24. The compound of Embodiment 23, wherein Xaa7 is an amino thiol residue of Cys, Cys-OH or Cys-NH₂, preferably of Cys-OH.

Embodiment 25. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24, preferably of any one of Embodiments 1, 2, 3 and 4, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro or Nmg, preferably an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln or Glu, preferably an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 26. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24, preferably any one of Embodiments 1, 2, 3 and 4, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 27. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24, preferably any one of Embodiments 1, 2, 3, and 4, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Glu,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 28. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 and 24, preferably any one of Embodiments 1, 2, 3 and 4, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Nmg,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 29. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27 and 28, wherein Yc is a a structure of

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wherein

R^c1is CH₂—R^c2or H,

CH₂—R^c2is a structure of formula (XIId) or of formula (XXIIb):

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wherein

Z is a chelator optionally comprising a linker

R^c4is H or methyl, and

u=1, 2, 3, 4 or 5.

Embodiment 30. The compound of Embodiment 29, wherein R^c2is a structure of formula (XIId)

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Embodiment 31. The compound of any one of Embodiments 29 and 30, wherein R^c2is a structure of formula (XIId)

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wherein

u=1, and

R^c4is H.

Embodiment 32. The compound of Embodiment 29, wherein R^c2is a structure of formula (XXIIc)

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Embodiment 33. The compound of any one of Embodiments 29, 30, 31 and 32, wherein Z is a chelator lacking a linker.

Embodiment 34. The compound of any one of Embodiments 29, 30, 31 and 32, wherein Z is a chelator comprising a linker.

Embodiment 35. The compound of Embodiment 34, wherein the linker is covalently linked to the chelator and covalently linked to the N-atom of the structure of formula (XIId)

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Embodiment 36. The compound of Embodiment 34, wherein the linker is covalently linked to the chelator and covalently linked to the N-atom of the structure of formula (XXIIc)

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Embodiment 37. The compound of any one of Embodiments 34, 35 and 36, wherein the linker is selected from the group consisting of Ttds and O2Oc.

Embodiment 38. The compound of Embodiment 37, wherein the linker is Ttds.

Embodiment 39. The compound of Embodiment 37, wherein the linker is O2Oc.

Embodiment 39. The compound of Embodiment 29, wherein R^c1is H.

Embodiment 40. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38 and 39, wherein an amino acid or a peptide is attached to Xaa7, wherein a majority of the amino acids of this peptide are charged or polar and the net charge of the peptide is −2, −1, 0, +1 or +2.

Embodiment 41. The compound of Embodiment 40, wherein an amino acid is attached to Xaa7.

Embodiment 42. The compound of Embodiment 41, wherein the amino acid attached to Xaa7 is selected from the group consisting of Asp, asp, Bal, Gly, Gab, Ser, Nmg, Bhf, Lys, Ape, Ttds and Bhk.

Embodiment 43. The compound of Embodiment 42, wherein the amino acid attached to Xaa7 is selected from the group consisting of Bhk, Ape and Lys.

Embodiment 44. The compound of Embodiment 43, wherein the amino acid attached to Xaa7 is Bhk.

Embodiment 45. The compound of any one of Embodiments 41, 42, 43 and 44, wherein a chelator Z is covalently attached to the amino acid attached to Xaa7.

Embodiment 46. The compound of Embodiment 45, wherein R^c1is H.

Embodiment 47. The compound of any one of Embodiments 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45 and 46, wherein Z is a chelator selected from the group consisting of ^99mTc(CO)₃-chelators, CB-TE2A, CHX-A″-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTAGA, DOTAM (also called TCMC), FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, DOTA, NODA-MPAA, NODAGA, NOTP, N_xS_4-x(N₄, N₂S₂, N₃S), NOPO, NOTA, Pycup, RESCA, Sarcophagine, TETA, THP, and TRAP.

Embodiment 48. The compound of Embodiment 47, wherein Z is a chelator selected form the group consisting of DOTAM, Macropa, PCTA, DOTA, N4Ac, NODAGA, NOPO, and NOTA.

Embodiment 49. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 and 48, wherein the compound is selected from the group consisting of

compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3940) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(N4Ac-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4533) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(N4Ac-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4534) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4560) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4564) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4565) of the following formula

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compound nBu-CAyl-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Bhk(N4Ac)—OH (3BP-4589) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4607) of the following formula

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and compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4621) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4723) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4724) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4768) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4778) of the following formula

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and compound nBu-CAyl-[Cys(tMeBn(NOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5210) of the following formula

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Embodiment 50. The compound of Embodiment 49, wherein the compound is selected from the group consisting of

compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3940) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4560) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4768) of the following formula

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Embodiment 51. The compound of any one of Embodiments 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49 and 50, wherein the chelator comprises a nuclide, preferably the nuclide is coordinatively bound by the chelator.

Embodiment 52. The compound of Embodiment 51, wherein the nuclide is a diagnostically active nuclide or a therapeutically active nuclide.

Embodiment 53. The compound of Embodiment 52, wherein the diagnostically active nuclide is a diagnostically active radionuclide.

Embodiment 54. The compound of Embodiment 53, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹Mn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb

Embodiment 55. The compound of Embodiment 54, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹¹In, and ²⁰³Pb.

Embodiment 56. The compound of Embodiment 52, wherein the therapeutically active nuclide is a therapeutically active radionuclide.

Embodiment 57. The compound of Embodiment 56, wherein the therapeutically active radionuclide is selected from the group consisting of ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb, ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th.

Embodiment 58. The compound of Embodiment 57, wherein the therapeutically active radionuclide is ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 59. A compound comprising a cyclic peptide

of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (11)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2eare each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein
- the substitution pattern of the aromatic group in formula (X) is ortho, meta or para, preferably meta,
- n=0 or 1,
- t=1 or 2,
- Y¹is C—H,
- Y²is C—R^c1,
- R^c1is CH₂—R^c2or H and
- R^c2is a structure of formula (XIId) or (XXIIc)

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wherein
- u=1,
- R^c4is H
- Z is a chelator optionally comprising a linker; and
- wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is R^a1—NH—C(O)—; wherein R^a1is (C₁-C₈)alkyl optionally substituted by up to two substituents each and independently selected from the group consisting of OH, F, COOH, (C₃-C₈)cycloalkyl, aryl, heteroaryl and (C₃-C₈)heterocycle, and wherein in (C₁-C₈)alkyl one of the —CH₂-groups is optionally replaced by —S— or —O—.

Embodiment 60. The compound of Embodiment 59, wherein R^c2is a structure of formula (XIId)

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wherein

u=1,

R^c4is H, and

Z is a chelator optionally comprising a linker.

Embodiment 61. The compound of Embodiment 60, wherein Z is a chelator lacking a linker.

Embodiment 62. The compound of Embodiment 60, wherein Z comprises a linker.

Embodiment 63. The compound of Embodiment 62, wherein the linker covalently links the chelator to the N-atom of the structure of formula (XIId).

Embodiment 64. The compound of any one of embodiments 62 to 63, wherein the linker is selected from the group consisting of Ttds, O2Oc and PEG6, preferably Ttds and O2Oc.

Embodiment 65. The compound of Embodiment 59, wherein R^c2is a structure of formula (XXIIc)

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wherein Z is a chelator optionally comprising a linker.

Embodiment 66. The compound of Embodiment 65, wherein Z is a chelator lacking a linker.

Embodiment 67. The compound of Embodiment 65, wherein Z comprises a linker.

Embodiment 68. The compound of Embodiment 67, wherein the linker covalently links the chelator to the N-atom of the structure of formula (XXIIc).

Embodiment 69. The compound of any one of embodiments 67 to 68, wherein the linker is selected from the group consisting of Ttds, O2Oc and PEG6, preferably the linker is selected from the group consisting of Ttds and O2Oc.

Embodiment 70. The compound of Embodiment 59, wherein R^c1is H.

Embodiment 71. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69 and 70, wherein R^a1is selected from the group consisting of C₃alkyl, C₄alkyl or C₅alkyl, each and independently optionally substituted by up to two substituents each and independently selected from the group consisting of OH, F, COOH, (C₃-C₈)cycloalkyl, aryl, heteroaryl and (C₃-C₈)heterocycle, and wherein in (C₁-C₈)alkyl one of the —CH₂-groups is optionally replaced by —S— or —O—.

Embodiment 72. The compound of Embodiment 71, wherein R^a1is selected from the group consisting of C₃alkyl, C₄alkyl or C₅alkyl.

Embodiment 73. The compound of any one of Embodiments 71 and 72, wherein R^a1is C₄alkyl.

Embodiment 74. The compound of Embodiment 73, wherein R^a1is n-butyl.

Embodiment 75. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73 and 74, wherein Xaa1 is a D-amino acid residue selected from the group consisting of cys, hcy and pen, or Xaa1 is an L-amino acid residue selected from the group consisting of Cys, Hcy and Pen.

Embodiment 76. The compound of Embodiment 75, wherein Xaa1 is Cys.

Embodiment 77. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 and 76, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro, Gly, Nmg and their derivatives.

Embodiment 78. The compound of Embodiment 77, wherein Xa2 is an amino acid residue selected from the group consisting of Pro and Nmg.

Embodiment 79. The compound of any one of Embodiments 77 and 78, wherein Xaa2 is an amino acid residue of Pro.

Embodiment 80. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78 and 79, wherein Xaa3 is an amino acid residue selected from the group consisting of Pro, Hyp, Tfp, Cfp, Dmp, Aze and Pip, and their derivatives.

Embodiment 81. The compound of Embodiment 80, wherein Xaa3 is an amino acid residue of Pro.

Embodiment 82. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 and 81, wherein Xaa4 is an amino acid residue selected from the group consisting of Thr, Hse, Asn, Gln and Ser, and their derivatives.

Embodiment 83. The compound of Embodiment 82, wherein Xaa4 is Thr.

Embodiment 84. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82 and 83, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu, and their derivatives.

Embodiment 85. The compound of Embodiment 84, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu.

Embodiment 86. The compound of Embodiment 85, wherein Xaa5 is an amino acid residue of Gln.

Embodiment 87. The compound of Embodiment 85, wherein Xaa5 is an amino acid residue of Glu.

Embodiment 88. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85 and 86, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each and independently selected from the group consisting of H, methyl, ethyl, propyl and isopropyl,
- R^6crepresents from 0 to 3 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand C₁-C₄alkyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0 or 1.

Embodiment 89. The compound of Embodiment 88, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each H
- R^6crepresents from 0 to 2 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand methyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0.

Embodiment 90. The compound of any one of Embodiments 88 to 89, wherein Xaa6 is an amino acid residue selected from the group consisting of Phe, Ocf, Ppa, Thi, 1Ni, Otf, and Mpa, and their derivatives.

Embodiment 91. The compound of Embodiment 90, wherein Xaa6 is an amino acid residue of Phe.

Embodiment 92. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80 and 81, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂Cysol, AET, Hcy, cys, cys-OH, cys-NH₂and hcy.

Embodiment 93. The compound of Embodiment 92, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂, Cysol and AET.

Embodiment 94. The compound of Embodiment 93, wherein Xaa7 is an amino thiol residue of Cys, Cys-OH or Cys-NH₂, preferably of Cys-OH.

Embodiment 95. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93 and 94, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro or Nmg, preferably an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln or Glu, preferably an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 96. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93 and 94, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 97. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93 and 94, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Glu,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 98. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93 and 94, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Nmg,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 99. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97 and 98, wherein an amino acid or a peptide is attached to Xaa7, wherein a majority of the amino acids of this peptide are charged or polar and the net charge of the peptide is −2, −1, 0, +1 or +2.

Embodiment 100. The compound of Embodiment 99, wherein an amino acid is attached to Xaa7.

Embodiment 101. The compound of Embodiment 100, wherein the amino acid attached to Xaa 7 is selected from the group consisting of Asp, asp, Bal, Gly, Gab, Ser, Nmg, Bhf, Lys, Ape, Ttds and Bhk.

Embodiment 102. The compound of Embodiment 101, wherein the amino acid attached to Xaa7 is selected from the group consisting of Bhk, Ape and Lys.

Embodiment 103. The compound of Embodiment 102, wherein the amino acid attached to Xaa7 is Bhk.

Embodiment 104. The compound of any one of Embodiments 100, 101, 192 and 103, wherein a chelator Z is covalently attached to the amino acid attached to Xaa7.

Embodiment 105. The compound of Embodiment 104, wherein R^c1is H.

Embodiment 106. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104 and 105, wherein Z is a chelator selected from the group consisting of ^99mTc(CO)₃-chelators, CB-TE2A, CHX-A″-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTAGA, DOTAM (also called TCMC), FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, DOTA, NODA-MPAA, NODAGA, NOTP, N_xS_4-x(N₄, N₂S₂, N₃S), NOPO, NOTA, Pycup, RESCA, Sarcophagine, TETA, THP, and TRAP.

Embodiment 107. The compound of Embodiment 106, wherein Z is a chelator selected form the group consisting of DOTAM, Macropa, PCTA, DOTA, N4Ac, NODAGA, NOPO, and NOTA.

Embodiment 108. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106 and 107, wherein the compound is selected from the group consisting of

compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3940) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(N4Ac-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4533) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(N4Ac-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4534) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4560) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4564) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4565) of the following formula

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compound nBu-CAyl-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Bhk(N4Ac)-OH (3BP-4589) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4607) of the following formula

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and compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4621) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4723) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4724) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4768) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4778) of the following formula

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and compound nBu-CAyl-[Cys(tMeBn(NOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5210) of the following formula

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Embodiment 109. The compound of Embodiment 108, wherein the compound is selected from the group consisting of

compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3940) of the following formula

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compound nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH2 (3BP-4560) of the following formula

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and compound nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4768) of the following formula

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Embodiment 110. The compound of any one of Embodiments 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108 and 109, wherein the chelator comprises a nuclide, preferably the nuclide is coordinatively bound by the chelator.

Embodiment 111. The compound of Embodiment 110, wherein the nuclide is a diagnostically active nuclide or a therapeutically active nuclide.

Embodiment 112. The compound of Embodiment 111, wherein the diagnostically active nuclide is a diagnostically active radionuclide.

Embodiment 113. The compound of Embodiment 112, wherein the diagnostically active radionuclide is selected from the group consisting of 18F, ⁴³Sc, ⁴⁴Sc, ⁵¹Mn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, 125I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and 203Pb.

Embodiment 114. The compound of Embodiment 113, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹¹In, and ²⁰³Pb.

Embodiment 115. The compound of Embodiment 111, wherein the therapeutically active nuclide is a therapeutically active radionuclide.

Embodiment 116. The compound of Embodiment 115, wherein the therapeutically active radionuclide is selected from the group consisting of ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb, ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th.

Embodiment 117. The compound of Embodiment 116, wherein the therapeutically active radionuclide is ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 118. A compound comprising a cyclic peptide

of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein

the substitution pattern of the aromatic group in formula (X) is meta,

n=0 or 1,

t=1 or 2,

Y¹is C—H or N,

Y²is C—R^c1,

R^c1is H;

wherein the N-terminal modification group A is an amino acid Aaa,

wherein

the amino acid Aaa is an L-amino acid residue of structure (XIV):

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wherein

R^a2is selected from the group consisting of (C₁-C₆)alkyl and modified (C₁-C₆)alkyl, wherein in modified (C₁-C₆)alkyl one —CH₂— group is replaced by —S— or —O—,

the amino acid Aaa is covalently attached to a linker, wherein the linker is covalently linked to a chelator Z, wherein the linker consists (a) of a first linker or (b) of a first linker and a second linker, wherein
- if the linker consists of the first linker, the first linker is covalently linked to the chelator and the amino acid Aaa, and
- if the first linker consists of a first linker and a second linker, the first linker is covalently linked to the amino acid Aaa and to the second linker, and the second linker is covalently linked to the chelator,
- the first linker is selected from the group consisting of Ttds and PEG6, preferably the first linker is Ttds,
- the second linker is selected from the group consisting of PPAc and PEG6, preferably the second linker is PPAc.

Embodiment 119. The compound of Embodiment 118, wherein R^a2is C₄alkyl.

Embodiment 120. The compound of any one of Embodiments 118 and 119, wherein the amino acid Aaa is a residue of Nle.

Embodiment 121. The compound of any one of Embodiments 118, 119 and 120, wherein Y¹is C—H.

Embodiment 122. The compound of any one of Embodiments 118, 119 and 120, wherein Y¹is N.

Embodiment 123. The compound of any one of Embodiments 118, 119, 120, 121 and 122, preferably any one of Embodiments 120 to 122, wherein the linker consists of a first linker, wherein the first linker is selected from the group consisting of Ttds and PEG6.

Embodiment 124. The compound of Embodiment 123, wherein the first linker is Ttds and, preferably the amino acid Aaa is a residue of Nle.

Embodiment 125. The compound of Embodiment 123, wherein the first linker is PEG6 and, preferably the amino acid Aaa is a residue of Nle.

Embodiment 126. The compound of any one of Embodiments 118, 119, 120, 121 and 122, preferably any one of Embodiments 120, 121 and 122, wherein the linker consists of a first linker and a second linker, wherein the first linker is selected from the group consisting of Ttds and PEG6, and the second linker is selected from the group consisting of PPAc and PEG6, preferably PPAc.

Embodiment 127. The compound of Embodiment 126, wherein the first linker is Ttds and the second linker is PPAc, preferably the amino acid Aaa is a residue of Nle.

Embodiment 128. The compound of Embodiment 126, wherein the first linker is Ttds and the second linker is PEG6, preferably the amino acid Aaa is a residue of Nle.

Embodiment 129. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127 and 128, wherein Xaa1 is a D-amino acid residue selected from the group consisting of cys, hcy and pen, or Xaa1 is an L-amino acid residue selected from the group consisting of Cys, Hcy and Pen.

Embodiment 130. The compound of Embodiment 129, wherein Xaa1 is Cys.

Embodiment 131. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129 and 130, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro, Gly, Nmg and their derivatives.

Embodiment 132. The compound of Embodiment 131, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro and Nmg.

Embodiment 133. The compound of any one of Embodiments 131 and 132, wherein Xaa2 is an amino acid residue of Pro.

Embodiment 134. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132 and 133, wherein Xaa3 is an amino acid residue selected from the group consisting of Pro, Hyp, Tfp, Cfp, Dmp, Aze and Pip, and their derivatives.

Embodiment 135. The compound of Embodiment 134, wherein Xaa3 is an amino acid residue of Pro.

Embodiment 136. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134 and 135, wherein Xaa4 is an amino acid residue selected from the group consisting of Thr, Hse, Asn, Gln and Ser, and their derivatives.

Embodiment 137. The compound of Embodiment 136, wherein Xaa4 is Thr.

Embodiment 138. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136 and 137, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu, and their derivatives.

Embodiment 139. The compound of Embodiment 138, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu.

Embodiment 140. The compound of Embodiment 139, wherein Xaa5 is an amino acid residue of Gln.

Embodiment 141. The compound of Embodiment 140, wherein Xaa5 is an amino acid residue of Glu.

Embodiment 142. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140 and 141, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each and independently selected from the group consisting of H, methyl, ethyl, propyl and isopropyl,
- R^6crepresents from 0 to 3 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR⁶d and C₁-C₄alkyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0 or 1.

Embodiment 143. The compound of Embodiment 142, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each H
- R^6crepresents from 0 to 2 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand methyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0.

Embodiment 144. The compound of any one of Embodiments 142 to 143, wherein Xaa6 is an amino acid residue selected from the group consisting of Phe, Ocf, Ppa, Thi, 1Ni, Otf, and Mpa, and their derivatives.

Embodiment 145. The compound of Embodiment 144, wherein Xaa6 is an amino acid residue of Phe.

Embodiment 146. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144 and 145, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂Cysol, AET, Hcy, cys, cys-OH, cys-NH₂and hcy.

Embodiment 147. The compound of Embodiment 146, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂, Cysol and AET.

Embodiment 148. The compound of Embodiment 147, wherein Xaa7 is an amino thiol residue of Cys, Cys-OH or Cys-NH₂, preferably of Cys-OH.

Embodiment 149. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147 and 148, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro or Nmg, preferably an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln or Glu, preferably an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 150. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147 and 148, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 151. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147 and 148, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Glu,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 152. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147 and 148, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Nmg,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 153. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151 and 152, wherein an amino acid or a peptide is attached to Xaa7, wherein a majority of the amino acids of this peptide are charged or polar and the net charge of the peptide is −2, −1, 0, +1 or +2.

Embodiment 154. The compound of Embodiment 153, wherein an amino acid is attached to Xaa7.

Embodiment 155. The compound of Embodiment 154, wherein the amino acid attached to Xaa 7 is selected from the group consisting of Asp, asp, Bal, Gly, Gab, Ser, Nmg, Bhf, Lys, Ape, Ttds and Bhk.

Embodiment 156. The compound of Embodiment 155, wherein the amino acid attached to Xaa7 is Bal or Asp.

Embodiment 157. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, wherein Z is a chelator selected from the group consisting of ^99mTc(CO)₃-chelators, CB-TE2A, CHX-A″-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTAGA, DOTAM (also called TCMC), FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, DOTA, NODA-MPAA, NODAGA, NOTP, N_xS_4-x(N₄, N₂S₂, N₃S), NOPO, NOTA, Pycup, RESCA, Sarcophagine, TETA, THP, and TRAP.

Embodiment 158. The compound of Embodiment 157, wherein Z is a chelator selected form the group consisting of DOTAM, Macropa, PCTA, DOTA, N4Ac, NODAGA, NOPO, and NOTA.

Embodiment 159. The compound of any one of Embodiments 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157 and 158, wherein the compound is selected from the group consisting of compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4541) of the following formula

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compound N4Ac-Ttds-Nle-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4549) of the following formula

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compound N4Ac-PEG6-Nle-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4550) of the following formula

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compound N4Ac-PEG6-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4551) of the following formula

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compound N4Ac-PEG6-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4552) of the following formula

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compound NODAGA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4713) of the following formula

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compound NODAGA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4714) of the following formula

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compound NODAGA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-NH₂(3BP-4743) of the following formula

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compound N4Ac-PEG6-Nle-[Cys(3Lut)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4773) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4774) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3Lut)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4775) of the following formula

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compound N4Ac-PEG6-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4780) of the following formula

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compound N4Ac-PPAc-PEG6-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4781) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4782) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Bal-OH (3BP-4784) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-OH (3BP-4785) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Bal-OH (3BP-4960) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-Bal-OH (3BP-4961) of the following formula

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and compound NOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5201) of the following formula

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Embodiment 160. The compound of Embodiment 159, wherein the compound is selected from the group consisting of

compound N4Ac-PPAc-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4541) of the following formula

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compound NODAGA-Ttds-Ne-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4713) of the following formula

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compound N4Ac-PPAc-Ttds-Nle-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-Bal-OH (3BP-4961) of the following formula

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and compound NOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5201) of the following formula

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Embodiment 161. The compound of any one of Embodiments 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159 and 160, wherein the chelator comprises a nuclide, preferably the nuclide is coordinatively bound by the chelator.

Embodiment 162. The compound of Embodiment 161, wherein the nuclide is a diagnostically active nuclide or a therapeutically active nuclide.

Embodiment 163. The compound of any one of Embodiments 162, wherein the diagnostically active nuclide is a diagnostically active radionuclide.

Embodiment 164. The compound of Embodiment 163, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹M, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, 125I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb.

Embodiment 165. The compound of Embodiment 164, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹¹In, and ²⁰³Pb.

Embodiment 166. The compound of any one of Embodiments 162, wherein the therapeutically active nuclide is a therapeutically active radionuclide.

Embodiment 167. The compound of Embodiment 166, wherein the therapeutically active radionuclide is selected from the group consisting of ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb, ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th.

Embodiment 168. The compound of Embodiment 167, wherein the therapeutically active radionuclide is ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 169. A compound comprising a cyclic peptide

of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (ITT), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein

the substitution pattern of the aromatic group in formula (X) is meta,

n=0 or 1,

t=1 or 2,

Y¹is C—H

Y²is C—R¹,

R^c1is CH₂—R^c2and

R^c2is a structure of formula (XIId)

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wherein
- u=1, 2, 3, 4, 5 or 6, preferably u=1,
- R^c4is H or methyl,
- Z is a chelator optionally comprising a linker, and
- wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is selected from the group consisting of R^a11—C(O)—, wherein R^a11is C₄alkyl or C₅alkyl, wherein in each and any one of C₄alkyl and C₅alkyl individually and independently one of the —CH₂-groups is optionally replaced by —O— or —S—.

Embodiment 170. The compound of Embodiment 169, wherein R^a11is C₅alkyl.

Embodiment 171. The compound of Embodiment 170, wherein R^a11is n-pentyl.

Embodiment 172. The compound of Embodiment 170, wherein R^a11is of structure (XXX).

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Embodiment 173. The compound of Embodiment 169, wherein R^a11is C₄alkyl.

Embodiment 174. The compound of Embodiment 173, wherein R^a11is n-butyl.

Embodiment 175. The compound of Embodiment 169, wherein R^11ais of structure (XXXI).

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Embodiment 176. The compound of Embodiment 169, wherein R^11ais of structure (XXXII)

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Embodiment 177. The compound of Embodiment 169, wherein R^11ais of structure (XXXIII).

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Embodiment 178. The compound of any one of Embodiments 169 to 177, wherein the chelator Z is covalently linked to the N atom of the structure of formula (XIId)

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Embodiment 179. The compound of Embodiment 170, wherein u=1.

Embodiment 180. The compound of any one of Embodiments 178 and 179, wherein R^c4is H.

Embodiment 181. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176 and 177, wherein the chelator Z comprises a linker.

Embodiment 182. The compound of Embodiment 181, wherein the linker is covalently linked to the chelator and covalently linked to the N atom of the structure of formula (XIId)

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Embodiment 183. The compound of Embodiment 182, wherein u=1.

Embodiment 184. The compound of any one of Embodiments 182 and 183, wherein R^c4is H.

Embodiment 185. The compound of any one of Embodiments 181, 182, 183 and 184, wherein the linker is selected from the group consisting of Ttds and O2Oc.

Embodiment 186. The compound of any one of Embodiments 181, 182, 183 and 184, wherein the linker is Ttds.

Embodiment 187. The compound of any one of Embodiments 181, 182, 183 and 184, wherein the linker is O2Oc.

Embodiment 188. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185 and 187, wherein Xaa1 is a D-amino acid residue selected from the group consisting of cys, hcy and pen, or Xaa1 is an L-amino acid residue selected from the group consisting of Cys, Hcy and Pen.

Embodiment 189. The compound of Embodiment 188, wherein Xaa1 is Cys.

Embodiment 190. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188 and 189, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro, Gly, Nmg and their derivatives.

Embodiment 191. The compound of Embodiment 190, wherein Xaa2 is an amino acid residue selected from the group consisting of Pro and Nmg.

Embodiment 192. The compound of any one of Embodiments 190 and 191, wherein Xaa2 is an amino acid residue of Pro.

Embodiment 193. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191 and 192, wherein Xaa3 is an amino acid residue selected from the group consisting of Pro, Hyp, Tfp, Cfp, Dmp, Aze and Pip, and their derivatives.

Embodiment 194. The compound of Embodiment 193, wherein Xaa3 is an amino acid residue of Pro.

Embodiment 195. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193 and 194, wherein Xaa4 is an amino acid residue selected from the group consisting of Thr, Hse, Asn, Gln and Ser, and their derivatives.

Embodiment 196. The compound of Embodiment 195, wherein Xaa4 is Thr.

Embodiment 197. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195 and 196, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu, and their derivatives.

Embodiment 198. The compound of Embodiment 197, wherein Xaa5 is an amino acid residue selected from the group consisting of Gln and Glu.

Embodiment 199. The compound of Embodiment 198, wherein Xaa5 is an amino acid residue of Gln.

Embodiment 200. The compound of Embodiment 199, wherein Xaa5 is an amino acid residue of Glu.

Embodiment 201. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199 and 200, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each and independently selected from the group consisting of H, methyl, ethyl, propyl and isopropyl,
- R⁶⁰represents from 0 to 3 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR⁶d and C₁-C₄alkyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0 or 1.

Embodiment 202. The compound of Embodiment 201, wherein Xaa6 is an amino acid residue of any one of formulae (VIIIa), (VIIIb), (VIIIc) and (VIIId):

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wherein
- R^6aand R^6bare each H
- R⁶⁰represents from 0 to 2 substituents, each such substituent being each and independently selected from the group consisting of Cl, F, Br, NO₂, NH₂, CN, CF₃, OH, OR^6dand methyl,
- R^6dis selected from the group consisting of methyl, ethyl, propyl, and isopropyl, and
- s is 0.

Embodiment 203. The compound of any one of Embodiments 201 to 202, wherein Xaa6 is an amino acid residue selected from the group consisting of Phe, Ocf, Ppa, Thi, 1Ni, Otf, and Mpa, and their derivatives.

Embodiment 204. The compound of Embodiment 203, wherein Xaa6 is an amino acid residue of Phe.

Embodiment 205. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203 and 204, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂Cysol, AET, Hcy, cys, cys-OH, cys-NH₂and hcy.

Embodiment 206. The compound of Embodiment 205, wherein Xaa7 is an amino thiol residue selected from the group consisting of Cys, Cys-OH, Cys-NH₂, Cysol and AET.

Embodiment 207. The compound of Embodiment 206, wherein Xaa7 is an amino thiol residue of Cys or Cys-NH₂, preferably of Cys-OH.

Embodiment 208. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206 and 207, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro or NmG, preferably an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln or Glu, preferably an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 209. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206 and 207, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 210. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206 and 207, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Pro,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Glu,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 211. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206 and 207, wherein

Xaa1 is an amino acid residue of Cys,
Xaa2 is an amino acid residue of Nmg,
Xaa3 is an amino acid residue of Pro,
Xaa4 is an amino acid residue of Thr,
Xaa5 is an amino acid residue of Gln,
Xaa6 is an amino acid residue of Phe, and
Xaa7 is an amino acid residue of Cys.

Embodiment 212. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210 and 211, wherein Z is a chelator selected from the group consisting of ^99mTc(CO)₃-chelators, CB-TE2A, CHX-A″-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTAGA, DOTAM (also called TCMC), FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, DOTA, NODA-MPAA, NODAGA, NOTP, N_xS_4-x(N₄, N₂S₂, N₃S), NOPO, NOTA, Pycup, RESCA, Sarcophagine, TETA, THP, and TRAP.

Embodiment 213. The compound of Embodiment 212, wherein Z is a chelator selected form the group consisting of DOTAM, Macropa, PCTA, DOTA, N4Ac, NODAGA, NOPO, and NOTA.

Embodiment 214. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212 and 213, wherein the compound is selected from the group consisting of compound iHex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3907) of the following formula

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compound Pent-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3910) of the following formula

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compound EtOPr-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3918) of the following formula

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compound MeOBut-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3937) of the following formula

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compound PrOAc-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3938) of the following formula

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compound nBu-COyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3941) of the following formula

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compound Hex-[Cys(tMeBn(DATA-Ttds-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4384) of the following formula

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compound Hex-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH (3BP-4695) of the following formula

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compound Hex-[Cys(tMeBn(NODAGA-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH₂(3BP-4708) of the following formula

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compound Hex-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-NH₂(3BP-4729) of the following formula

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compound Hex-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Glu-Phe-Cys]-OH(3BP-4818) of the following formula

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compound Hex-[Cys(tMeBn(AcPCTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5273) of the following formula

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compound Hex-[Cys(tMeBn(LSC-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5288) of the following formula

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and compound Hex-[Cys(tMeBn(DOTAM-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-5323) of the following formula

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Embodiment 215. The compound of any one of Embodiments 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213 and 214, wherein the chelator comprises a nuclide, preferably the nuclide is coordinatively bound by the chelator.

Embodiment 216. The compound of Embodiment 215, wherein the nuclide is a diagnostically active nuclide or a therapeutically active nuclide.

Embodiment 217. The compound of Embodiment 216, wherein the diagnostically active nuclide is a diagnostically active radionuclide.

Embodiment 218. The compound of Embodiment 217, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹Mn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb.

Embodiment 219. The compound of Embodiment 218, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹¹In, and ²⁰³Pb.

Embodiment 220. The compound of Embodiment 216, wherein the therapeutically active nuclide is a therapeutically active radionuclide.

Embodiment 221. The compound of Embodiment 220, wherein the therapeutically active radionuclide is selected from the group consisting of ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb, ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th.

Embodiment 222. The compound of Embodiment 221, wherein the therapeutically active radionuclide is ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 223. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221 and 222, wherein the compound interacts with a fibroblast activation protein (FAP), preferably with human FAP having an amino acid sequence of SEQ ID NO: 1 or a homolog thereof, wherein the amino acid sequence of the homolog has an identity of at least 85% to the amino acid sequence of SEQ ID NO: 1.

Embodiment 224. The compound of Embodiment 223, wherein the compound is an inhibitor of the fibroblast activation protein (FAP).

Embodiment 225. The compound of any one of Embodiments 1 to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223 and 224, wherein the compound has a pIC₅₀value for human FAP of SEQ ID NO: 1 of ≥6.0, preferably of ≥7.0, and most preferably of ≥8.0.

Embodiment 226. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, preferably of any one of Embodiments 5 to 55, 110 to 114, 161 to 165 and 215 to 219, for use in a method for the diagnosis of a disease.

Embodiment 227. The compound for use of Embodiment 226, wherein the disease is a disease involving fibroblast activation protein (FAP), preferably upregulated expression of fibroblast activation protein (FAP).

Embodiment 228. The compound for use of any one of Embodiments 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226 and 227, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 229. The compound for use of any one of Embodiments 226, 227 and 228, wherein the disease is a neoplasm, preferably a cancer or tumor.

Embodiment 230. The compound for use of Embodiment 229, wherein the neoplasm, cancer, and tumor are each and individually selected from the group comprising a solid tumor, an epithelial tumor, 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 carcinoma, sarcoma, squamous cell carcinoma, and thyroid cancer.

Embodiment 231. The compound for use of Embodiment 230, wherein the neoplasm, cancer, and tumor are each and 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 232. The compound for use of any one of Embodiments 226, 227 and 228, wherein the disease is selected from the groups comprising inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease.

Embodiment 233. The compound for use of Embodiment 232, wherein the disease is an inflammatory disease.

Embodiment 234. The compound for use of Embodiment 233, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis.

Embodiment 235. The compound for use of Embodiment 232, wherein the disease is a cardiovascular disease.

Embodiment 236. The compound for use of Embodiment 235, wherein the disease is a cardiovascular disease involving atherosclerotic plaques.

Embodiment 237. The compound for use of Embodiment 236, wherein the disease is an atherosclerotic pathology caused by rupture of plaques, acute coronary syndrome, myocardial infarction, thrombosis, or vessel occlusion.

Embodiment 238. The compound for use of Embodiment 232, wherein the disease is a fibrotic disease.

Embodiment 239. The compound for use of Embodiment 238, wherein the disease is selected form the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis.

Embodiment 240. The compound for use of any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238 and 239, wherein the compound comprises a diagnostically active nuclide, preferably a diagnostically active radionuclide.

Embodiment 241. The compound for use of Embodiment 240, wherein the diagnostically active nuclide is selected from the group comprising ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹n, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb. preferably ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹In, and ²⁰³Pb.

Embodiment 242. The compound for use of any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240 and 241, wherein the method for the diagnosis is an imaging method.

Embodiment 243. The compound for use of Embodiment 242, wherein the imaging method is selected from the group consisting of scintigraphy, Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET).

Embodiment 244. The compound for use of any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242 and 243, wherein the method comprises the administration of a diagnostically effective amount of the compound to a subject, preferably to a mammal, wherein the mammal is selected from the group comprising man, companion animals, pets, and livestock, more preferably the subject is selected from the group comprising man, dog, cat, horse, and cow, and most preferably the subject is a human being.

Embodiment 245. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, preferably any one of Embodiments 51, 52, 56 to 58, 110, 111, 115 to 117, 161, 162, 166 to 168, 215, 216 and 220 to 222, for use in a method for the treatment of a disease.

Embodiment 246. The compound for use of Embodiment 245, wherein the disease is a disease involving fibroblast activation protein (FAP), preferably upregulated expression of fibroblast activation protein (FAP).

Embodiment 247. The compound for use of any one of Embodiments 245 to 246, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 248. The compound for use of any one of Embodiments 245, 246 and 247, wherein the disease is a neoplasm, preferably a cancer or tumor.

Embodiment 249. The compound for use of Embodiment 248, wherein the neoplasm, cancer, and tumor are each and individually selected from the group comprising a solid tumor, an epithelial tumor, 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 carcinoma, sarcoma, squamous cell carcinoma, and thyroid cancer.

Embodiment 250. The compound for use of Embodiment 249, wherein the neoplasm, cancer, and tumor are each and 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 251. The compound for use of any one of Embodiments 245, 246 and 247, wherein the disease is selected from the groups comprising inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease.

Embodiment 252. The compound for use of Embodiment 251, wherein the disease is an inflammatory disease.

Embodiment 253. The compound for use of Embodiment 252, wherein the disease is atherosclerosis, arthritis, or rheumatoid arthritis.

Embodiment 254. The compound for use of Embodiment 251, wherein the disease is a cardiovascular disease.

Embodiment 255. The compound for use of Embodiment 254, wherein the diseases is a cardiovascular disease involving atherosclerotic plaques.

Embodiment 256. The compound for use of Embodiment 255, wherein the diseases is an atherosclerotic pathology caused by rupture of plaques, acute coronary syndrome, myocardial infarction, thrombosis, or vessel occlusion.

Embodiment 257. The compound for use of Embodiment 251, wherein the disease is a fibrotic disease.

Embodiment 258. The compound for use of Embodiment 257, wherein the disease is selected form the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis.

Embodiment 259. The compound for use of any one of Embodiments 245, 246, 247 and 248, wherein the compound comprises a therapeutically active nuclide, preferably a therapeutically active radionuclide.

Embodiment 260. The compound for use of Embodiment 259, wherein the therapeutically active nuclide is selected from the group comprising ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th, preferably ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 261. The compound for use of any one of Embodiments 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259 and 260, wherein the method comprises the administration of a therapeutically effective amount of the compound to a subject, preferably to a mammal, wherein the mammal is selected from the group comprising man, companion animals, pets, and livestock, more preferably the subject is selected from the group comprising man, dog, cat, horse, and cow, and most preferably the subject is a human being.

Embodiment 262. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, for use in a method for the identification of a subject, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, wherein the method for the identification of a subject comprises carrying out a method of diagnosis using the compound of any one of Embodiments, preferably a method for the diagnosis of a disease as described in any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243 and 244.

Embodiment 263. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, for use in a method for the selection of a subject from a group of subjects, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, wherein the method for the selection of a subject from a group of subjects comprises carrying out a method of diagnosis using the compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, preferably a method for the diagnosis of a disease as described in any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243 and 244.

Embodiment 264. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, for use in a method for the stratification of a group of subjects into subjects which are likely to respond to a treatment of a disease, and into subjects which are not likely to respond to a treatment of a disease, wherein the method for the stratification of a group of subjects comprises carrying out a method of diagnosis using the compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, preferably a method for the diagnosis of a disease as described in any one of Embodiments 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243 and 244.

Embodiment 265. The compound for use of any one of Embodiments 262, 263 and 264, wherein the disease is a disease involving fibroblast activation protein (FAP), preferably upregulated expression of fibroblast activation protein (FAP).

Embodiment 266. The compound for use of any one of Embodiments 262, 263, 264 and 265, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 267. The compound for use of any one of Embodiments 262, 263, 264, 165 and 266, wherein the disease is a neoplasm, preferably a cancer or tumor.

Embodiment 268. The compound for use of Embodiment 267, wherein the neoplasm, cancer, and tumor are each and individually selected from the group comprising a solid tumor, an epithelial tumor, 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 carcinoma, sarcoma, squamous cell carcinoma, and thyroid cancer.

Embodiment 269. The compound for use of Embodiment 268, wherein the neoplasm, cancer, and tumor are each and 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 270. The compound for use of any one of Embodiments 262, 263, 264, 265 and 266, wherein the disease is selected from the groups comprising inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease.

Embodiment 271. The compound for use of Embodiment 270, wherein the disease is an inflammatory disease.

Embodiment 272. The compound for use of Embodiment 271, wherein the disease is atherosclerosis, arthritis or rheumatoid arthritis.

Embodiment 273. The compound for use of Embodiment 272, wherein the disease is a cardiovascular disease.

Embodiment 274. The compound for use of Embodiment 273, wherein the disease is a cardiovascular disease involving atherosclerotic plaques.

Embodiment 275. The compound for use of Embodiment 274, wherein the disease is an atherosclerotic pathology caused by rupture of plaques, acute coronary syndrome, myocardial infarction, thrombosis, or vessel occlusion.

Embodiment 276. The compound for use of Embodiment 270, wherein the disease is a fibrotic disease.

Embodiment 277. The compound for use of Embodiment 276, wherein the disease is selected from the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis.

Embodiment 278. The compound for use of any one of Embodiments 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276 and 277, wherein the method of diagnosis is an imaging method.

Embodiment 279. The compound for use of Embodiment 278 wherein the imaging method is selected from the group comprising scintigraphy, Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET).

Embodiment 280. The compound for use of any one of Embodiments 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278 and 279, wherein the compound comprises a diagnostically active nuclide, preferably a diagnostically active radionuclide.

Embodiment 281. The compound for use of Embodiment 280, wherein the diagnostically active nuclide is selected from the group comprising ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹Mn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb. preferably ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹In, and ²⁰³Pb.

Embodiment 282. The compound of any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, for use in a method for delivering an effector to fibroblast activation protein (FAP), preferably human fibroblast activation protein (FAP), wherein the effector is selected from the group comprising a diagnostically active agent and a therapeutically active agent.

Embodiment 283. The compound for use of Embodiment 282, wherein the effector is selected from the group comprising a diagnostically active nuclide and a therapeutically active nuclide.

Embodiment 284. The compound for use of Embodiment 283, wherein the diagnostically active nuclide is a diagnostically active radionuclide.

Embodiment 285. The compound for use of Embodiment 284, wherein the diagnostically active radionuclide is selected from the group consisting of ¹⁸F, ⁴³Sc, ⁴⁴Sc, ⁵¹Nn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁷⁶Br, ⁷⁷Br, ⁸⁶Y, ⁸⁹Zr, ⁹⁴mTc, ^99mTc, ¹¹¹In, ¹²³I, ¹²⁴I, ¹²⁵I, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, and ²⁰³Pb. preferably ¹⁸F, ⁶⁸Ga, ^99mTc, ¹¹¹In, and ²⁰³Pb.

Embodiment 286. The compound for use of any one of Embodiments 282, 283, 284 and 285, wherein the fibroblast activation protein (FAP) is expressed by a cell, preferably a fibroblast, a mesenchymal stem cell, smooth muscle cell, a cell of epithelial origin, or an 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 showing upregulated expression of fibroblast activation protein (FAP).

Embodiment 287. The compound for use of Embodiment 286, wherein the cell is contained in or part of a tissue, preferably a diseased tissue of a subject suffering from a disease.

Embodiment 288. The compound for use of Embodiment 287, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 289. The compound for use of any one of Embodiments 287 to 288, wherein the disease is a neoplasm, preferably a cancer or tumor.

Embodiment 290. The compound for use of Embodiment 289, wherein the neoplasm, cancer, and tumor are each and individually selected from the group comprising a solid tumor, an epithelial tumor, 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 carcinoma, sarcoma, squamous cell carcinoma, and thyroid cancer.

Embodiment 291. The compound for use of Embodiment 290, wherein the neoplasm, cancer, and tumor are each and 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 292. The compound for use of any one of Embodiments 287 to 288, wherein the disease is selected from the groups comprising inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease.

Embodiment 293. The compound for use of Embodiment 292, wherein the disease is an inflammatory disease.

Embodiment 294. The compound for use of Embodiment 293, wherein the disease is atherosclerosis, arthritis or rheumatoid arthritis.

Embodiment 295. The compound for use of Embodiment 292, wherein the disease is a cardiovascular disease.

Embodiment 296. The compound for use of Embodiment 295, wherein the diseases is a cardiovascular disease involving atherosclerotic plaques.

Embodiment 297. The compound for use of Embodiment 296, wherein the disease is an atherosclerotic pathology caused by rupture of plaques, acute coronary syndrome, myocardial infarction, thrombosis, or vessel occlusion.

Embodiment 298. The compound for use of Embodiment 292, wherein the disease is a fibrotic disease.

Embodiment 299. The compound for use of Embodiment 298, wherein the disease is selected form the group comprising idiopathic pulmonary fibrosis, Crohn's disease, and liver fibrosis.

Embodiment 300. The compound for use of Embodiment 283, wherein the therapeutically active nuclide is a therapeutically active radionuclide.

Embodiment 301. The compound for use of Embodiment 300, wherein the therapeutically active radionuclide is selected from the group consisting of ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹³¹I, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹¹At, ²¹²Pb, ²¹³Bi, ²²³Ra, ²²⁴Ra, ²²⁵Ac, ²²⁶Th, and ²²⁷Th, preferably ⁹⁰Y, ¹⁷⁷Lu, ²¹²Pb, and ²²⁵Ac.

Embodiment 302. The compound for use of any one of Embodiment 300 to 301, wherein the fibroblast activation protein (FAP) is expressed by a cell, preferably a fibroblast, a mesenchymal stem cell, smooth muscle cell, a cell of epithelial origin, or an 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 showing upregulated expression of fibroblast activation protein (FAP).

Embodiment 303. The compound for use of Embodiment 302, wherein the cell is contained in or part of a tissue, preferably a diseased tissue of a subject suffering from a disease.

Embodiment 304. The compound for use of Embodiment 303, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 305. The compound for use of any one of Embodiments 302, 303 and 304, wherein the disease is a neoplasm, preferably a cancer or tumor.

Embodiment 306. The compound for use of Embodiment 305, wherein the neoplasm, cancer, and tumor are each and individually selected from the group comprising a solid tumor, an epithelial tumor, 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 carcinoma, sarcoma, squamous cell carcinoma, and thyroid cancer.

Embodiment 307. A composition, preferably a pharmaceutical composition, wherein the composition comprises a compound according to any one of Embodiment 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225 and a pharmaceutically acceptable excipient.

Embodiment 308. The composition of Embodiment 307 for use in any method as defined in any of the preceding claims.

Embodiment 309. A method for the diagnosis of a disease in a subject, wherein the method comprises administering to the subject a diagnostically effective amount of a compound according to any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225.

Embodiment 310. The method of Embodiment 309, wherein the compound comprises a diagnostically active agent, whereby the agent is preferably a radionuclide.

Embodiment 311. A method for the treatment of a disease in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a compound according to any one of Embodiment 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225.

Embodiment 312. The method of Embodiment 311, wherein the compound comprises a therapeutically active agent, whereby the agent is preferably a radionuclide.

Embodiment 313. The method of any one of Embodiments 309, 310, 311 and 312, wherein the disease is a disease involving fibroblast activation protein (FAP), preferably upregulated expression of fibroblast activation protein (FAP).

Embodiment 314. The method of any one of Embodiments 309, 310, 311, 312 and 313, wherein the disease involves cells showing upregulated expression of fibroblast activation protein (FAP), preferably diseased tissue containing cells showing upregulated expression of fibroblast activation protein (FAP), more preferably disease involving tumor associated fibroblasts.

Embodiment 315. The method of any one of Embodiments 309, 310, 311, 312, 313 and 314, wherein the disease is selected from the groups comprising neoplasms, preferably cancers or tumors, and inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease.

Embodiment 316. A kit comprising a compound according to any one of Embodiments 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224 and 225, one or more optional excipient(s) and optionally one or more device(s), whereby the device(s) is/are selected from the group comprising a labeling device, a purification device, a handling device, a radioprotection device, an analytical device or an administration device.

Embodiment 317. The kit of Embodiment 316 for use in any method as defined in any of the preceding Embodiments.

More specifically, the problem underlying the present invention is solved in a first aspect by a compound comprising a cyclic peptide

of formula (I)

embedded image

and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen
  - atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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- wherein
- R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H,
- wherein R^7band R^7care each and independently (C₁-C₄)alkyl and
- t is 1 or 2;
- Yc is a structure of formula (X)

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- linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein
- the substitution pattern of the aromatic group in formula (X) is ortho, meta or para,
- n=0 or 1,
- t=1 or 2,
- Y¹is C—H or N,
- Y²is N or C—R^c1,
- R^c1is H or CH₂—R^c2and
- R^c2is a structure of formula (XI), (XII) or (XXII)

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wherein
- R^c3and R^c4are each and independently selected from the group consisting of H and (C₁-C₄)alkyl and
- u=1, 2, 3, 4, 5 or 6,
- x and y are each and independently 1, 2 or 3, and
- X═O or S

wherein in formulae (XI) and (XXII) one of the nitrogen atoms is attached to —CH₂— of R^c1and in formula (XII) —X— is attached to —CH₂— of R^c1; and

wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is R^a1—NH—C(O)—; wherein R^a1is selected from the group consisting of C₃alkyl, C₄alkyl or C₅alkyl, each and independently optionally substituted by up to two substituents each and independently selected from the group consisting of OH, F, COOH, (C₃-C₈)cycloalkyl, aryl, heteroaryl and (C₃-C₈)heterocycle, and wherein in (C₁-C₈)alkyl one of the —CH₂-groups is optionally replaced by —S— or —O—.

More specifically, the problem underlying the present invention is solved in a second aspect by a compound comprising a compound comprising a cyclic peptide

of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen
  - atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein
- the substitution pattern of the aromatic group in formula (X) is ortho, meta or para, preferably meta,
- n=0 or 1,
- t=1 or 2,
- Y¹is C—H,
- Y²is C—R^c1,
- R^c1is CH₂—R^c2or H and
- R^c2is a structure of formula (XIId) or (XXIIc)

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wherein
- u=1,
- R^c4is H
- Z is a chelator optionally comprising a linker; and
- wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is R^a1—NH—C(O)—; wherein R^a1is (C₁-C₈)alkyl optionally substituted by up to two substituents each and independently selected from the group consisting of OH, F, COOH, (C₃-C₈)cycloalkyl, aryl, heteroaryl and (C₃-C₈)heterocycle, and wherein in (C₁-C₈)alkyl one of the —CH₂-groups is optionally replaced by —S— or —O—.

More specifically, the problem underlying the present invention is solved in a third aspect by a compound comprising a compound comprising a cyclic peptide of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen
  - atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein

the substitution pattern of the aromatic group in formula (X) is meta,

n=0 or 1,

t=1 or 2,

Y¹is C—H or N,

Y²is C—R^c1,

R^c1is H;

wherein the N-terminal modification group A is an amino acid Aaa,

wherein

the amino acid Aaa is an L-amino acid residue of structure (XIV):

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wherein

R^a2is selected from the group consisting of (C₁-C₆)alkyl and modified (C₁-C₆)alkyl, wherein in modified (C₁-C₆)alkyl one —CH₂— group is replaced by —S— or —O—,

the amino acid Aaa is covalently attached to a linker, wherein the linker is covalently linked to a chelator Z, wherein the linker consists (a) of a first linker or (b) of a first linker and a second linker, wherein
- if the linker consists of the first linker, the first linker is covalently linked to the chelator and the amino acid Aaa, and
- if the first linker consists of a first linker and a second linker, the first linker is covalently linked to the amino acid Aaa and to the second linker, and the second linker is covalently linked to the chelator,
- the first linker is selected from the group consisting of Ttds and PEG6, preferably the first linker is Ttds,
- the second linker is selected from the group consisting of PPAc and PEG6, preferably the second linker is PPAc.

More specifically, the problem underlying the present invention is solved in a fourth aspect by a compound comprising a compound comprising a cyclic peptide

of formula (I)

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and an N-terminal modification group A attached to Xaa1,

wherein
- the peptide sequence is drawn from left to right in N to C-terminal direction,
- Xaa1 is a residue of an amino acid of formula (II)

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- wherein
  - R^1ais —NH—
  - R^1bis H or CH₃,
  - n=0 or 1,
  - the N-terminal modification group A is covalently attached to the nitrogen atom of Xaa1,
  - the carbonyl group of Xaa1 is covalently attached to the nitrogen of Xaa2,
  - and the sulfur atom of Xaa1 is covalently attached as thioether to Yc;
- Xaa2 is a residue of an amino acid of formula (III), (IV) or (XX)

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- wherein
  - R^2a, R^2band R^2care each and independently selected from the group consisting of (C₁-C₂)alkyl and H, wherein said (C₁-C₂)alkyl maybe substituted by a substituent selected from the group consisting of OH, NH₂, halogen, (C₅-C₇)cycloalkyl,
  - p=0, 1 or 2
  - v=1 or 2
  - w=1, 2 or 3 and
  - the amino acid of formula (IV) maybe substituted by one or two substituents selected from the group consisting of methyl, OH, NH₂and F, at indicated ring positions 3 and 4;
- Xaa3 is a residue of an amino acid of formula (V) or (XX)

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- wherein
  - X³is selected from the group consisting of CH₂, CF₂, CH—R^3b, S, O and NH,
  - p=1 or 2
  - v=1 or 2
  - w=1, 2 or 3,
  - R^3ais H, methyl, OH, NH₂or F,
  - R^3bis methyl, OH, NH₂or F;
- Xaa4 is a residue of an amino acid of formula (VI)

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- - wherein
  - R^4ais selected from the group consisting of H, OH, COOH, CONH₂, X⁴and —NH—CO—X⁴, wherein X⁴is selected from the group consisting of (C₁-C₆)alkyl, (C₅-C₆)aryl and (C₅-C₆)heteroaryl, and X⁴may be substituted by one or two substituents selected from the group consisting of methyl, CONH₂, halogen, NH₂and OH;
  - q=1, 2 or 3, wherein optionally, one or two hydrogens of said one, two, or three CH₂-groups are each and individually substituted by methyl, ethyl, (C₅-C₆)aryl or (C₅-C₆)heteroaryl,
  - R^4bis methyl or H;
- Xaa5 is a residue of an amino acid of structure (VII)

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- wherein
  - R⁵is selected from the group of OH and NH₂, and
  - r=1, 2 or 3;
- Xaa6 is an amino acid selected from the group consisting of an aromatic L-α-amino acid and a heteroaromatic L-α-amino acid;
- Xaa7 is a residue of an amino thiol or an amino acid of formula (IX),

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wherein

R^7ais —CO—, —COOH, —CONH₂, —CH₂—OH, —(CO)—NH—R^7b, —(CO)—(NR^7c)—R^7bor H, wherein

R^7band R^7care each and independently (C₁-C₄)alkyl and

t is 1 or 2;
- Yc is a structure of formula (X)

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linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages thus forming a cyclic structure of formula (XXI)

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wherein

the substitution pattern of the aromatic group in formula (X) is meta,

n=0 or 1,

t=1 or 2,

Y¹is C—H

Y²is C—R^c1,

R^c1is CH₂—R^c2and

R^c2is a structure of formula (XIId)

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wherein
- u=1, 2, 3, 4, 5 or 6, preferably u=1,
- R^c4is H or methyl,
- Z is a chelator optionally comprising a linker, and
- wherein the N-terminal modification group A is a blocking group Abl, wherein the blocking group Abl is selected from the group consisting of R^a11—C(O)—, wherein R^a11is C₄alkyl or C₅alkyl, wherein in each and any one of C₄alkyl and C₅alkyl individually and independently one of the —CH₂-groups is optionally replaced by —O— or —S—.

More specifically, the problem underlying the present invention is solved in a fifth aspect by the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, for use in a method for the diagnosis of a disease.

More specifically, the problem underlying the present invention is solved in a sixth aspect by the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, for use in a method for the treatment of a disease.

More specifically, the problem underlying the present invention is solved in a seventh aspect by the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, for use in a method for the identification of a subject, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, wherein the method for the identification of a subject comprises carrying out a method of diagnosis using the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof.

More specifically, the problem underlying the present invention is solved in an eighth aspect by the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, for use in a method for the selection of a subject from a group of subjects, wherein the subject is likely to respond or likely not to respond to a treatment of a disease, wherein the method for the selection of a subject from a group of subjects comprises carrying out a method of diagnosis using the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof.

More specifically, the problem underlying the present invention is solved in a ninth aspect by the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, for use in a method for the stratification of a group of subjects into subjects which are likely to respond to a treatment of a disease, and into subjects which are not likely to respond to a treatment of a disease, wherein the method for the stratification of a group of subjects comprises carrying out a method of diagnosis using the compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof.

More specifically, the problem underlying the present invention is solved in a tenth aspect by a composition, preferably a pharmaceutical composition, wherein the composition comprises a compound according to the first aspect second aspect, third aspect, and fourth aspect, including any embodiment thereof, and a pharmaceutically acceptable excipient.

More specifically, the problem underlying the present invention is solved in an eleventh aspect by a method for the diagnosis of a disease in a subject, wherein the method comprises administering to the subject a diagnostically effective amount of a compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof.

More specifically, the problem underlying the present invention is solved in a 12^thaspect by a method for the treatment of a disease in a subject, wherein the method comprises administering to the subject a therapeutically effective amount of a compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof.

More specifically, the problem underlying the present invention is solved in a 13^thaspect by a kit comprising a compound according to the first aspect, second aspect, third aspect, and fourth aspect, including any embodiment thereof, one or more optional excipient(s) and optionally one or more device(s), whereby the device(s) is/are selected from the group comprising a labeling device, a purification device, a handling device, a radioprotection device, an analytical device or an administration device.

It will be acknowledged by a person skilled in the art that a or the compound of the 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.

It will be acknowledged by a person skilled in the art that a or the method of the 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.

It will be acknowledged by a person skilled in the art that a or the composition of the 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.

It will be acknowledged by a person skilled in the art that a or the kit of the 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.

It will be acknowledged that in connection with the present invention, any embodiments of any aspect of the invention may also be an embodiment of any other aspect of the invention, including any embodiment thereof.

The present invention is based on the surprising finding of the present inventors that the compound of the invention and more specifically the cyclic peptide thereof provides for a highly specific binding of a compound comprising such cyclic peptide to fibroblast activation protein (FAP), since FAP-specific cyclic peptide-based inhibitors with nanomolar affinity have not been described so far.

Furthermore, the present invention is based on the surprising finding that a chelator, either directly or indirectly, i.e., using a linker, may be attached to said cyclic peptide at three different positions. The first position is Yc having a structure of formula (X) which links the S atom of Xaa1 and the S atom of Xaa7 thus forming two thioether linkages; the second position is Aaa attached to Xaa1 of the cyclic peptide of formula (I), and the third position is an amino acid or a peptide attached to Xaa7. Surprisingly, the attachment of such chelator does not significantly affect the binding of the compound of the invention to FAP and, respectively, the inhibiting characteristics of the compound of the present invention on FAP. In one embodiment, the present invention relates to the cyclic peptide of formula (I) where a chelator (Z group) is attached at only one of the first, second, or third position as defined above. It is also within the present invention that the chelator is attached to the cyclic peptide of formula (I) at any combination of the first, second, and third position as defined above. More specifically, the present invention also relates to compound of formula (I) where a Z group is attached to both the first and the second position as defined above, a compound of formula (I) where a Z group is attached to both the first and the third position as defined above, a compound of formula (I) where a Z group is attached to both the second and the third position as defined above, and a compound of formula (I) where a Z group is attached to the first, the second and the third position as defined above. These compounds comprising two or three Z groups may be realized in any embodiment of the present invention as disclosed herein.

Finally, the present inventors have found that the compounds of the invention are surprisingly stable in blood plasma and are surprisingly useful as imaging agents and efficacious in shrinking tumors.

The expression alkyl as preferably used herein refers each and individually to a saturated, straight-chain or branched hydrocarbon group and is usually accompanied by a qualifier which specifies the number of carbon atoms it may contain. For example the expression (C₁-C₆)alkyl means each and individually any of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methyl-butyl, 1-ethyl-propyl, 3-methyl-butyl, 1,2-dimethyl-propyl, 2-methyl-butyl, 1,1-dimethyl-propyl, 2,2-dimethylpropyl, n-hexyl, 1,1-dimethyl-butyl and any other isoform of alkyl groups containing six saturated carbon atoms.

In an embodiment and as preferably used herein, (C₁-C₂)alkyl means each and individually any of methyl and ethyl.

In an embodiment and as preferably used herein, (C₁-C₃)alkyl means each and individually any of methyl, ethyl, n-propyl and isopropyl.

In an embodiment and as preferably used herein, (C₁-C₄)alkyl means each and individually any of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

In an embodiment and as preferably used herein, (C₁-C₆)alkyl means each and individually any of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 2-methyl-butyl, 3-methyl-butyl, 3-pentyl, 3-methyl-but-2-yl, 2-methyl-but-2-yl, 2,2-dimethylpropyl, n-hexyl, 2-hexyl, 2-methyl-pentyl, 3-methyl-pentyl, 4-methyl-pentyl, 3-hexyl, 2-ethyl-butyl, 2-methyl-pent-2-yl, 2,2-dimethyl-butyl, 3,3-dimethyl-butyl, 3-methyl-pent-2-yl, 4-methyl-pent-2-yl, 2,3-dimethyl-butyl, 3-methyl-pent-3-yl, 2-methyl-pent-3-yl, 2,3-dimethyl-but-2-yl and 3,3-dimethyl-but-2-yl.

In an embodiment and as preferably used herein, (C₁-C₈)alkyl refers to a saturated or unsaturated, straight-chain or branched hydrocarbon group having from 1 to 8 carbon atoms.

Representative (C₁-C₈)alkyl groups include, but are not limited to, any of methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 2-methyl-butyl, 3-methyl-butyl, 3-pentyl, 3-methyl-but-2-yl, 2-methyl-but-2-yl, 2,2-dimethylpropyl, n-hexyl, 2-hexyl, 2-methyl-pentyl, 3-methyl-pentyl, 4-methyl-pentyl, 3-hexyl, 2-ethyl-butyl, 2-methyl-pent-2-yl, 2,2-dimethyl-butyl, 3,3-dimethyl-butyl, 3-methyl-pent-2-yl, 4-methyl-pent-2-yl, 2,3-dimethyl-butyl, 3-methyl-pent-3-yl, 2-methyl-pent-3-yl, 2,3-dimethyl-but-2-yl, 3,3-dimethyl-but-2-yl, n-heptyl, 2-heptyl, 2-methyl-hexyl, 3-methyl-hexyl, 4-methyl-hexyl, 5-methyl-hexyl, 3-heptyl, 2-ethyl-pentyl, 3-ethyl-pentyl, 4-heptyl, 2-methyl-hex-2-yl, 2,2-dimethyl-pentyl, 3,3-dimethyl-pentyl, 4,4-dimethyl-pentyl, 3-methyl-hex-2-yl, 4-methyl-hex-2-yl, 5-methyl-hex-2-yl, 2,3-dimethyl-pentyl, 2,4-dimethyl-pentyl, 3,4-dimethyl-pentyl, 3-methyl-hex-3-yl, 2-ethyl-2-methyl-butyl, 4-methyl-hex-3-yl, 5-methyl-hex-3-yl, 2-ethyl-3-methyl-butyl, 2,3-dimethyl-pent-2-yl, 2,4-dimethyl-pent-2-yl, 3,3-dimethyl-pent-2-yl, 4,4-dimethyl-pent-2-yl, 2,2,3-trimethyl-butyl, 2,3,3-trimethyl-butyl, 2,3,3-trimethyl-but-2-yl, n-octyl, 2-octyl, 2-methyl-heptyl, 3-methyl-heptyl, 4-methyl-heptyl, 5-methyl-heptyl, 6-methyl-heptyl, 3-octyl, 2-ethyl-hexyl, 3-ethyl-hexyl, 4-ethyl-hexyl, 4-octyl, 2-propyl-pentyl, 2-methyl-hept-2-yl, 2,2-dimethyl-hexyl, 3,3-dimethyl-hexyl, 4,4-dimethyl-hexyl, 5,5-dimethyl-hexyl, 3-methyl-hept-2-yl, 4-methyl-hept-2-yl, 5-methyl-hept-2-yl, 6-methyl-hept-2-yl, 2,3-dimethyl-hex-1-yl, 2,4-dimethyl-hex-1-yl, 2,5-dimethyl-hex-1-yl, 3,4-dimethyl-hex-1-yl, 3,5-dimethyl-hex-1-yl, 3,5-dimethyl-hex-1-yl, 3-methyl-hept-3-yl, 2-ethyl-2-methyl-1-yl, 3-ethyl-3-methyl-1-yl, 4-methyl-hept-3-yl, 5-methyl-hept-3-yl, 6-methyl-hept-3-yl, 2-ethyl-3-methyl-pentyl, 2-ethyl-4-methyl-pentyl, 3-ethyl-4-methyl-pentyl, 2,3-dimethyl-hex-2-yl, 2,4-dimethyl-hex-2-yl, 2,5-dimethyl-hex-2-yl, 3,3-dimethyl-hex-2-yl, 3,4-dimethyl-hex-2-yl, 3,5-dimethyl-hex-2-yl, 4,4-dimethyl-hex-2-yl, 4,5-dimethyl-hex-2-yl, 5,5-dimethyl-hex-2-yl, 2,2,3-trimethyl-pentyl, 2,2,4-trimethyl-pentyl, 2,3,3-trimethyl-pentyl, 2,3,4-trimethyl-pentyl, 2,4,4-trimethyl-pentyl, 3,3,4-trimethyl-pentyl, 3,4,4-trimethyl-pentyl, 2,3,3-trimethyl-pent-2-yl, 2,3,4-trimethyl-pent-2-yl, 2,4,4-trimethyl-pent-2-yl, 3,4,4-trimethyl-pent-2-yl, 2,2,3,3-tetramethyl-butyl, 3,4-dimethyl-hex-3-yl, 3,5-dimethyl-hex-3-yl, 4,4-dimethyl-hex-3-yl, 4,5-dimethyl-hex-3-yl, 5,5-dimethyl-hex-3-yl, 3-ethyl-3-methyl-pent-2-yl, 3-ethyl-4-methyl-pent-2-yl, 3-ethyl-hex-3-yl, 2,2-diethyl-butyl, 3-ethyl-3-methyl-pentyl, 4-ethyl-hex-3-yl, 5-methyl-hept-3-yl, 2-ethyl-3-methyl-pentyl, 4-methyl-hept-4-yl, 3-methyl-hept-4-yl, 2-methyl-hept-4-yl, 3-ethyl-hex-2-yl, 2-ethyl-2-methyl-pentyl, 2-isopropyl-pentyl, 2,2-dimethyl-hex-3-yl, 2,2,4-trimethyl-pent-3-yl and 2-ethyl-3-methyl-pentyl. A (C₁-C₈)alkyl group can be unsubstituted or substituted with one or more groups, including, but not limited to, (C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

The expression alkylidene as preferably used herein refers to a saturated straight chain or branched hydrocarbon group wherein two points of substitution are specified. Simple alkyl chains wherein the two points of substitutions are in a maximal distance to each other like methane-1,1-diyl, ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl and pentane-1,5-diyl are also referred to as methylene (which is also referred to as methane-1,1-diyl), ethylene (which is also referred to as ethane-1,2-diyl), propylene (which is also referred to as propane-1,3-diyl), butylene (which is also referred to as butane-1,4-diyl) and pentylene (which is also referred to as pentane-1,5-diyl).

In an embodiment and as preferably used herein, (C₁-C₁₀)alkylidene means each and individually any of methylene, ethane-1,2-diyl, propane-1,3-diyl, propane-1,2-diyl, butane-1,4-diyl, butane-1,3-diyl, butane-1,2-diyl, 2-methyl-propane-1,2-diyl, 2-methyl-propane-1,3-diyl, pentane-1,5-diyl, pentane-1,4-diyl, pentane-1,3-diyl, pentane-1,2-diyl, pentane-2,3-diyl, pentane-2,4-diyl, any other isomer with 5 carbon atoms, hexane-1,6-diyl, any other isomer with 6 carbon atoms, heptane-1,7-diyl, any other isomer with 7 carbon atoms, octane-1,8-diyl, any other isomer with 8 carbon atoms, nonane-1,9-diyl, any other isomer with 9 carbon atoms, decane-1,10-diyl and any other isomer with 10 carbon atoms, preferably (C₁-C₁₀) alkylidene means each and individually any of methylene, ethane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl, pentane-1,5-diyl, hexane-1,6-diyl, heptane-1,7-diyl, octane-1,8-diyl, nonane-1,9-diyl and decane-1,10-diyl. A (C₁-C₁₀)alkylidene group can be unsubstituted or substituted with one or more groups, including, but not limited to, (C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment and as preferably used herein, (C₃-C₈)cycloalkyl means each and individually any of cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

In an embodiment and as preferably used herein, (C₅-C₇)cycloalkyl means each and individually any of cyclopentyl, cyclohexyl and cycloheptyl.

In an embodiment and as preferably used herein, (C₃-C₈)carbocycle refers to a 3-, 4-, 5-, 6-, 7- or 8-membered saturated or unsaturated non-aromatic carbocyclic ring. Representative (C3-C8)carbocycles include, but are not limited to, any of -cyclopropyl, -cyclobutyl, -cyclopentyl, -cyclopentadienyl, -cyclohexyl, -cyclohexenyl, -1,3-cyclohexadienyl, -1,4-cyclohexadienyl, -cycloheptyl, -1,3-cycloheptadienyl, -1,3,5-cycloheptatrienyl, -cyclooctyl, and -cylooctadienyl. A (C₃-C₈)carbocycle group can be unsubstituted or substituted with one or more groups, including, but not limited to, (C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment and as preferably used herein, (C3-C8)carbocyclo refers to a (C₃-C₈)carbocycle group defined above wherein one of the carbocycles group hydrogen atoms is replaced with a bond.

In an embodiment and as preferably used herein, “aryl” refers to a carbocyclic aromatic group. Examples of aryl groups include, but are not limited to, phenyl, naphthyl and anthracenyl.

In an embodiment and as preferably used herein, (C₅-C₆)aryl refers to a 5 or 6 carbon atom comprising carbocyclic aromatic group. A carbocyclic aromatic group can be unsubstituted or substituted with one or more groups including, but not limited to, —(C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment and as preferably used herein, “heteroaryl” refers to a heterocyclic aromatic group. Examples of heteroaryl groups include, but are not limited to, furane, thiophene, pyridine, pyrimidine, benzothiophene, benzofurane and quinoline.

In an embodiment and as preferably used herein, (C₅-C₆)heteroaryl refers to a heterocyclic aromatic group consisting of 5 or 6 ring atoms wherein at least one atom is different from carbon, preferably nitrogen, sulfur or oxygen. A heterocyclic aromatic group can be unsubstituted or substituted with one or more groups including, but not limited to, —(C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment and as preferably used herein, (C₃-C₈)heterocyclo refers to a (C₃-C₈)heterocycle group defined above wherein one of the carbocycles group hydrogen atoms is replaced with a bond. A (C₃-C₈)heterocyclo can be unsubstituted or substituted with up to six groups including, (C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment and as preferably used herein, arylene refers to an aryl group which has two covalent bonds and can be in the ortho, meta, or para configurations as shown in the following structures:

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in which the phenyl group can be unsubstituted or substituted with four groups, including, but not limited to, (C₁-C₈)alkyl, —O—[(C₁-C₈)alkyl], -aryl, —CO—R′, —O—CO—R′, —CO—OR′, —CO—NH₂, —CO—NHR′, —CO—NR′₂, —NH—CO—R′, —SO₂—R′, —SO—R′, —OH, -halogen, —N₃, —NH₂, —NHR′, —NR′₂and —CN; where each R′ is independently selected from —(C₁-C₈)alkyl and aryl.

In an embodiment of each and any aspect, including any embodiment thereof, any S atom which can be oxidized, preferably S atoms of thioether groups, is present as —S—, —S(O)— or —S(O2)- or a mixture thereof.

In an embodiment and as preferably used herein atoms with unspecified atomic mass numbers in any structural formula or in any passage of the instant specification including the claims are either of unspecified isotopic composition, naturally occurring mixtures of isotopes or individual isotopes. This applies in particular to carbon, oxygen, nitrogen, sulfur, phosphorus, halogens and metal atoms, including but not limited to C, O, N, S, F, P, Cl, Br, At, Sc, Cr, Mn, Co, Fe, Cu, Ga, Sr, Zr, Y, Mo, Tc, Ru, Rh, Pd, Pt, Ag, In, Sb, Sn, Te, I, Pr, Pm, Dy, Sm, Gd, Tb, Ho, Dy, Er, Yb, Tm, Lu, Sn, Re, Rd, Os, Ir, Au, Pb, Bi, Po, Fr, Ra, Ac, Th and Fm.

In an embodiment and as preferably used herein, a chelator is a compound which is capable of forming a chelate, whereby a chelate is a compound, preferably a cyclic compound where a metal or a moiety having an electron gap or a lone pair of electrons participates in the formation of the ring. More preferably, a chelator is this kind of compound where a single ligand occupies more than one coordination site at a central atom.

In an embodiment and as preferably used herein, a diagnostically active compound is a compound which is suitable for or useful in the diagnosis of a disease.

In an embodiment and as preferably used herein, a diagnostic agent or a diagnostically active agent is a compound which is suitable for or useful in the diagnosis of a disease.

In an embodiment and as preferably used herein, a therapeutically active compound is a compound which is suitable for or useful in the treatment of a disease.

In an embodiment and as preferably used herein, a therapeutic agent or a therapeutically active agent is a compound which is suitable for or useful in the treatment of a disease.

In an embodiment and as preferably used herein, a theragnostically active compound is a compound which is suitable for or useful in both the diagnosis and therapy of a disease.

In an embodiment and as preferably used herein, a theragnostic agent or a theragnostically active agent is a compound which is suitable for or useful in both the diagnosis and therapy of a disease.

In an embodiment and as preferably used herein, theragonstics is a method for the combined diagnosis and therapy of a disease; preferably, the combined diagnostically and therapeutically active compounds used in theragnostics are radiolabeled.

In an embodiment and as preferably used herein, treatment of a disease is treatment and/or prevention of a disease.

In an embodiment and as preferably used herein, a disease involving FAP is a disease where cells including but not limited to fibroblasts expressing, preferably in an upregulated manner, FAP and tissue either expressing FAP or containing or comprising cells such as fibroblasts, preferably expressing FAP in an upregulated manner respectively, are either a or the cause for the disease and/or the symptoms of the disease, or are part of the pathology underlying the disease. A preferred FAP-expressing cell is a cancer associated fibroblast (CAF). In an embodiment of the disease, preferably when used in connection with the treatment, treating and/or therapy of the disease, affecting the cells, the tissue and pathology, respectively, results in cure, treatment or amelioration of the disease and/or the symptoms of the disease. In an embodiment of the disease, preferably when used in connection with the diagnosis and/or diagnosing of the disease, labeling of the FAP-expressing cells and/or of the FAP-expressing tissue allows discriminating or distinguishing said cells and/or said tissue from healthy or FAP-non-expressing cells and/or healthy or FAP non-expressing tissue. More preferably such discrimination or distinction forms the basis for said diagnosis and diagnosing, respectively. In an embodiment thereof, labeling means the interaction of a detectable label either directly or indirectly with the FAP-expressing cells and/or with the FAP-expressing tissue or tissue 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 FAP.

In an embodiment and as preferably used herein, a target cell is a cell which is expressing FAP and is a or the cause for a disease and/or the symptoms of a disease, or is part of the pathology underlying a disease.

In an embodiment and as preferably used herein, a non-target cell is a cell which is either not expressing FAP and/or is not a or the cause for a disease and/or the symptoms of a disease, or is part of the pathology underlying a disease.

In an embodiment and as preferably used herein, a neoplasm is an abnormal new growth of cells. The cells in a neoplasm grow more rapidly than normal cells and will continue to grow if not treated. A neoplasm may be benign or malignant.

In an embodiment and as preferably used herein, a tumor is a mass lesion that may be benign or malignant.

In an embodiment and as preferably used herein, a cancer is a malignant neoplasm.

In an embodiment and as preferably used herein, a linkage is an attachment of two atoms of two independent moieties. A preferred linkage is a chemical bond or a plurality of chemical bonds. More preferably a chemical bond is a covalent bond or a plurality of chemical bonds. Most preferably the linkage is a covalent bond or a coordinate bond. As preferably used herein, an embodiment of a coordinate bond is a bond or group of bonds as realized when a metal is bound by a chelator. Depending on the type of atoms linked and their atomic environment different types of linkages are created. These types of linkage are defined by the type of atom arrangements created by the linkage. For instance, the linking of a moiety comprising an amine with a moiety comprising a carboxylic acid leads to a linkage named amide (which is also referred to as amide linkage, —CO—N—, —N—CO—). It will be acknowledged by a person skilled in the art that this and the following examples of creating linkages are only prototypical examples and are by no means limiting the scope of the instant application. It will be acknowledged by a person in the art that the linking of a moiety comprising an isothiocyanate with a moiety comprising an amine leads to thiourea (which is also referred to as a thiourea linkage, —N—CS—N—), and linking of a moiety comprising a C atom with a moiety comprising a thiol-group (—C—SH) leads to thioether (which is also referred to as a thioether linkage, —C—S—C—). A non-limiting list of linkages as preferably used in connection with the chelator and linker of the invention and their characteristic type of atom arrangement is presented Table 2.

TABLE 2

Linkage
Characteristic atom arrangement

Amide

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Sulfonamide

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Urea

Thioether

Disulfide

Ether

Ester

Carbamate

Thiourea

Triazole

Pyrazine

Dihydro-pyrazine

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Examples of reactive groups which, in some embodiments of the invention, are used in the formation of linkages between the chelator and linker or directly between the chelator and the compound of the invention are summarized in Table 3. It will, however, be understood by a person skilled in the art that neither the linkages which may be realized in embodiments for the formation of the conjugates of the invention are limited to the ones of Table 3 nor the reactive groups forming such linkages.

TABLE 3

first reactive group
second reactive group
(type of) linkage

amino
carboxylic acid
amide

amino
activated carboxylic acid
amide

carboxylic acid
amino
amide

sulfhydryl
Michael acceptor (e.g. Maleimide)
thioether

bromo
sulfhydryl
thioether

isothiocyanate
amino
thiourea

hydroxyl
carboxylic acid
ester

azide
alkyne
triazole

sulfhydryl
sulfhydryl
disulfide

sulfhydryl
2-Pyridine-disulfide
disulfide

isocyanate
amino
carbamate

bromo
hydroxy
ether

The following are reactive groups and functionalities which are utilized or amenable of forming linkages between moieties or structures as used in embodiments of the conjugate of the invention:

Primary or secondary amino, carboxylic acid, activated carboxylic acid, chloro, bromo, iodo, sulfhydryl, hydroxyl, sulfonic acid, activated sulfonic acid, sulfonic acid esters like mesylate or tosylate, Michael acceptors, strained alkenes like trans cyclooctene, isocyanate, isothiocyanate, azide, alkyne and tetrazine.

As preferably used herein, the term “activated carboxylic acid” refers to a carboxylic acid group with the general formula —CO—X, wherein X is a leaving group. For example, activated forms of a carboxylic acid group may include, but are not limited to, acyl chlorides, symmetrical or unsymmetrical anhydrides, and esters. In some embodiments, the activated carboxylic acid group is an ester with pentafluorophenol, nitrophenol, benzotriazole, azabenzotriazole, thiophenol or N-hydroxysuccinimide (NHS) as leaving group.

As preferably used herein, the term “activated sulfonic acid” refers to a sulfonic acid group with the general formula —SO₂—X, wherein X is a leaving group. For example, activated forms of a sulfonic acid may include, but are not limited to, sulfonyl chlorides or sulfonic acid anhydrides. In some embodiments, the activated sulfonic acid group is sulfonylchloride with chloride as leaving group.

In an embodiment and as preferably used herein the term “mediating a linkage” means that a linkage or a type of linkage is established, preferably a linkage between two moieties. In a preferred embodiment the linkage and the type of linkage is as defined herein.

To the extent it is referred in the instant application to a range indicated by a lower integer and a higher integer such as, for example, 1-4, such range is a representation of the lower integer, the higher integer and any integer between the lower integer and the higher integer. Insofar, the range is actually an individualized disclosure of said integer. In said example, the range of 1-4 thus means 1, 2, 3 and 4.

Compounds of the invention typically contain amino acid sequences as provided herein. Conventional amino acids, also referred to as natural amino acids are identified according to their standard three-letter codes and one-letter abbreviations, as set forth in Table 4.

TABLE 4

Conventional amino acids and their abbreviations

3-letter
1-letter

Amino acid
abbreviation
abbreviation

Alanine
Ala
A

Arginine
Arg
R

Asparagine
Asn
N

Aspartic acid
Asp
D

Cysteine
Cys
C

Glutamic acid
Glu
E

Glutamine
Gln
Q

Glycine
Gly
G

Histidine
His
H

Isoleucine
Ile
I

Leucine
Leu
L

Lysine
Lys
K

Methionine
Met
M

Phenylalanine
Phe
F

Proline
Pro
P

Serine
Ser
S

Threonine
Thr
T

Tryptophan
Trp
W

Tyrosine
Tyr
Y

Valine
Val
V

Non-conventional amino acids, also referred to as non-natural amino acids, are any kind of non-oligomeric compound which comprises an amino group and a carboxylic group and is not a conventional amino acid.

Examples of non-conventional amino acids and other building blocks as used for the construction compounds of the invention are identified according to their abbreviation or name found in Table 5. The structures of some building blocks are depicted with an exemplary reagent for introducing the building block into the peptide (e.g., as carboxylic acid like) or these building blocks are shown as residue which is completely attached to another structure like a peptide or amino acid. The structures of the amino acids are shown as explicit amino acids and not as residues of the amino acids how they are presented after implementation in the peptide sequence. Some larger chemical moieties consisting of more than one moiety are also shown for the reason of clarity.

TABLE 5

Abbreviation, name and structure of non-natural amino-acid and other building blocks

and chemical moieties

Abbreviation
Name
Structure

1Ni
3-(1-naphthyl)alanine

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2Lut
2,6-lutidylidene (derived from 2,6-lutidine)

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2Ni
3-(2-naphthyl)alanine

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3Lut
3,5-lutidylidene (derived from 3,5-lutidine)

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3MeBn
3-Methylbenzylidene

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4Amc
4-trans- Aminomethylcyclohexane carboxylic acid/ Tranexamic acid

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4Ap
(2S,4S)-4-Amino- pyrrolidine-2-carboxylic acid

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4Dfp
4,4-Difluoroproline

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4Pya
2-(Pyridin-4-yl)acetic acid

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4Tfp
4-trans-Fluoroproline

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Aad
(S)-Homo glutamic acid

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Abu
(S)-2-Amino-butyric acid

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AET
2-Aminoethanethiol

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AF488
Alexa Fluor 488 Dye

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Ahx
6-Amino-hexanoic acid

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Aib
2-Amino-isobutyric acid

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Aic
2-Aminoindane-2- carboxylic acid

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Alloc-
Allyloxycarbonyl-

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Amf
(S)-α-Methyl-phenylalanine

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APAc
2-(4-(Amino)piperidin-1- yl)acetic acid

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Ape
1,5-Diaminopentane

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Ape(DOTA)
4-[[(5-Amino-pentylcarbamoyl)- methyl]-7,10-bis- carboxymethyl- 1,4,7,10tetraaza-cyclododec-1- yl]-acetic acid

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ATTO488
Atto 488 Dye

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Ava
5-Amino-pentanoic acid

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Aze
(S)-Azetidine-2-carboxylic acid

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Bal
β-Alanine

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Bhf
(S)-β-Homophenylalanine

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Bhk
(S)-β-Homolysine

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Bio
D(+)-Biotin

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Bip
(S)-Biphenylalanine

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Bulloc-
But-3-enyloxycarbonyl-

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Cfp
4-cis-Fluoro proline

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Chg
(S)-Cyclohexylglycine

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Chex
Cyclohexyl carboxylic acid

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Chy
(2S,4S)-4-Hydroxy-pyrrolidine- 2-carboxylic acid

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Cit
(S)-Citrulline

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Cmp
4-Carboxymethyl-piperidine

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Cp
Cyclopentyl carboxylic acid

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Cpentyl-CAyl-
Cyclopentylaminocarbonyl-

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Cpp
trans-3- Azabicyclo[3.1.0]hexane-2- carboxylic acid

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CuDOTA
DOTA complexing Copper

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Cy5SO3
Cy5 dye (mono SO3)

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Cya
(R)-Cysteic acid

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Cys(2Lut)

Cys(3Lut)

Cys(3MeBn)

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Cys(tMeBn (DOTA-AET))

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Cys(tMeBn (DOTA-PP))

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Cys(tMeBn (H-AET))

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Cys(tMeBn (H-PP))

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Cys-NH₂
Cysteine modified as carboxamide

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Cys-OH
Cysteine with free carboxylic acid

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Cysol
(R)-Cysteinol

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Dab
(S)-2,4-Diaminobutyric acid

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Dap
(S)-2,3-Diaminopropionic acid

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DATA
(6-Pentanoic acid)-6- (amino)methy-1,4- diazepinetriacetate

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Dmp
(S)-5,5-Dimethyl-proline

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DOTA
1,4,7,10- Tetraazacyclododecane- 1,4,7,10-tetraacetic acid

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DTPA
Diethylenetriaminepentaacetic acid

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DTPA2
Diethylenetriaminepentaacetic acid

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DTPABzl
(S)-2-(4-Aminobenzyl)- diethylenetriaminepentaacetic acid

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Eay
(2S,4S)-4-phenyl-pyrrolidine-2- carboxylic acid

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Efa
N-[2-(2-Amino-ethanesulfonyl)- ethyl]-succinamic acid

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Egd
(S)-ω,ω-Dimethyl-arginine

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EtOPr
3-Ethoxy-propionic acid

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EuDOTA
DOTA complexing Europium

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Fur
Tetrahydrofuran-3-carboxylic acid

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Gab
γ-Aminobutyric acid

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GaDOTA
DOTA complexing Gallium

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GaNODAGA
NODAGA complexing Gallium

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Ghg
(S)-γ-Hydroxy-glutamic acid

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Glu(AGLU)

Glutar
Glutaric acid

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H2NSO2-But
4-Sulfamoylbutyric acid

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H3p
(2S,3S)-3-hydroxy-pyrrolidine-2- carboxylic acid

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Har
(S)-Homoarginine

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HBED
N,N-bis(2- hydroxybenzyl)ethylenediamine- N,N-diacetic acid

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Hci
(S)-Homocitrulline

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Hcy
(S)-Homocysteine

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hcy
(R)-Homocysteine

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Hex
Hexanoic acid

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Hex-
Hexanoyl-

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Hfe
(S)-Homophenylalanine

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Hga
(S)-Homoglutamic acid

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Hgl
(S)-n-Hexylglycin

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Hle
(S)-Homoleucine

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Hse
(S)-Homoserine

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Hth
(S)-Homothreonine

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Hym
(2S,4R)-4-Methyoxy-pyrrolidine- 2-carboxylic acid

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Hyn
Hex-5-ynoic acid

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HYNIC
Hydrazinonicotinic acid

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Hyp
(2S,4R)-4-Hydroxy-pyrrolidine- 2-carboxylic acid

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iHex
4-Methyl-pentanoic acid

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InDOTA
DOTA complexing Indium

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Inp
Isonipecotic acid

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LuDOTA
DOTA complexing Lutetium

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Mamb
3-Aminomethyl-benzoic acid

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MeOBut
4-Methoxy-butyric acid

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Moo
(S)-Methionine sulfone

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Mpa
3-Pyridyl-alanine

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N4Ac
6-Carboxy-1,4,8,11- tetraazaundecane; a N4-chelator

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nBu-CAyl-
n-Butylaminocarbonyl-

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nBu-COyl-
n-Butyloxycarbonyl-

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Nle
(S)-Norleucine

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nle
(R)-Norleucine

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Nleu
N-(Isobutyl)-glycine

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Nlys
4-Aminobutyl-glycine

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Nma
(S)-N-Methyl-alanine

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nma
(R)-N-Methyl-alanine

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Nmc
(R)-N-Methyl-cysteine

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Nme
(S)-N-Methyl-glutamic acid

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Nmf
(S)-N-Methyl-phenylalanine

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Nmg
N-Methyl-glycine

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NODAGA
1,4,7-Triazacyclononane,1- glutaric acid-4,7-acetic acid

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NOPO
3-(((4,7- Bis((hydroxy(hydroxymethyl) phosphoryl)methyl)-1,4,7- triazonan-1- yl) methyl) (hydroxy) phosphoryl) propanoic acid

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NOTA
2,2′2″-(1,4,7- Triazacyclononane-1,4,7- triyl)triacetic acid

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Nphe
N-Benzyl glycine

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Nva
(S)-Norvaline

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O2Oc
3,6-Dioxaoctanoic acid

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Ocf
(S)-2-Chloro-phenylalanine

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Oct
Octanoic acid

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Oic
(S)-Octahydroindolecarboxylic acid

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Omr
(S)-ω-Methyl-arginine

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Opc
(S)-N-(Pyrazinylcarbonyl)-ornithine

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Orn
(S)-Ornithine

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Otf
(S)-2-Trifluoromethyl-phenylalanine

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Peet
Pent-4-enoic acid

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Pent
Pentanoic acid

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PP
Piperazinyliden

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Pamb
4-Aminomethyl-benzoic acid

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Pcf
(S)-4-Chloro-phenylalanine

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PCTA
3,6,9,15- Tetraazabicyclo[9.3.1] pentadeca-1 (15),11,13-triene-3,6,9- triacetic acid

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PEG12

PEG6

Pen
(R)-Penicillamine

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pen
(S)-Penicillamine

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PentylNH-urea

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Pentyl-SO2-
Pentyl sulfonyl

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Php
3-Phenylpropionic acid

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Pip
(S)-Piperidine-2-carboxylic acid

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Ppa
(S)-4-Pyridyl-alanine

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PPAc
4-Carboxymethyl piperazine

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PrOAc
Propoxy-acetic acid

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Pyn
Pent-4-ynoic acid

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ReON4Ac
Oxo-rhenium(V) complex of N4Ac

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Rni
(R)-Nipecotic acid

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Rth
(R)-Tetrahydrofuran-2- carboxylic acid

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SAc
Mercapto acetic acid

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Sni
(S)-Nipecotic acid

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Spa
3-Mercaptopropionic acid

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Sth
(S)-Tetrahydrofuran- 2-carboxylic acid

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-Succ-
-Succinimide-

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Tap
(2S,4S)-4-Amino-pyrrolidine- 2-carboxylic acid

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Tfp
(2S,4S)-4-Fluoro-pyrrolidine- 2-carboxylic acid

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Thi
(S)-β-(2-Thienyl)-alanine

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Tic
(S)-1,2,3,4- Tetrahydroisoquinoline-3- carboxylic acid

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tMeBn
1,3,5-Trimethylbenzyliden

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tMeBn(H-AET)

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tMeBn(H-PP)

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Ttds
1,13-Diamino-4,7,10- trioxatridecan-succinamic acid

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ZnDOTA
Zink complex of DOTA

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The amino acid sequences of the peptides provided herein are depicted in typical peptide sequence format, as would be understood by the ordinary skilled artisan. For example, the three-letter code of a conventional amino acid, or the code for a non-conventional amino acid or the abbreviations for additional building blocks, indicates the presence of the amino acid or building block in a specified position within the peptide sequence. The code for each amino acid or building block is connected to the code for the next and/or previous amino acid or building block in the sequence by a hyphen which (typically represents an amide linkage).

Where an amino acid contains more than one amino and/or carboxy group all orientations of this amino acid are in principle possible, but in α-amino acid the utilization of the α-amino and the α-carboxy group is preferred and otherwise preferred orientations are explicitly specified.

For amino acids, in their abbreviations the first letter indicates the stereochemistry of the C-α-atom if applicable. For example, a capital first letter indicates that the L-form of the amino acid is present in the peptide sequence, while a lower case first letter indicating that the D-form of the correspondent amino acid is present in the peptide sequence.

In an embodiment and as preferably used herein, an aromatic L-α-amino acid is any kind of L-α-amino acid which comprises an aryl group.

In an embodiment and as preferably used herein, a heteroaromatic L-α-amino acid is any kind of L-α-amino acid which comprises a heteroaryl group.

Those skilled in the art will recognize if a stereocenter exists in the compounds disclosed herein irrespective thereof whether such stereocenter is part of an amino acid moiety or any other part or moiety of the compound of the invention. Accordingly, the present invention includes both possible stereoisomers and includes not only racemic compounds but the individual enantiomers and/or diastereomers as well. When a compound is desired as a single enantiomer or diastereomer, it may be obtained by stereospecific synthesis or by resolution of the final product or any convenient intermediate. Resolution of the final product, an intermediate, or a starting material may be affected by any suitable method known in the art. See, for example, “Stereochemistry of Organic Compounds” by E. L. Eliel, S. H. Wilen, and L. N. Mander (Wiley-Interscience, 1994).

In the present application, the structural formula of the compound represents a certain isomer for convenience in some cases, but the present invention includes all isomers, such as geometrical isomers, optical isomers based on an asymmetrical carbon, stereoisomers, tautomers, and the like. In the present specification, the structural formula of the compound represents a certain isomer for convenience in some cases, but the present invention includes all isomers, such as geometrical isomers, optical isomers based on an asymmetrical carbon, stereoisomers, tautomers, and the like.

Unless indicated to the contrary, the amino acid sequences are presented herein in N- to C-terminus direction.

Derivatives of the amino acids constituting the peptides of the invention may be as set forth in Table 6. In any embodiment, one or more amino acids of the compounds of the invention are substituted with a derivative of the corresponding preferred amino acids.

TABLE 6

Exemplary derivatives of preferred amino acids contained in the compound of the invention

Amino Acid
Exemplary derivatives

Ala
Aib, Bal, Abu, Gly, Nva, Nle

Cys
Hcy, Nmc

Asp
Glu, Asn, Gln, Cya

Glu
Asp, Asn, Gln, Cya, Homoglutamic acid, γ-Hydroxy-glutamic acid,

Phe
Hfe, Phg, Bhf, Thi, Bta, Bromophenylalanine, Iodophenylalanine,

Chlorophenylalanine, Methylphenylalanine, Nitrophenylalanine, Tyr, Trp,

Naphthylalanine, Trifluoromethylphenylalanine

Gly
Ala, ala, Nmg

Nmg
Pro, Ala, ala, Gly, Nma, nma

His
1-Methylhistidine, 3-Methylhistidine, Thi

Ile
Leu, Val, Hle, Nva, Nle, Chg

Lys
Arg, Dab, Dap, Har, Egd, Omr, Hci, Cit

Leu
Ile, Val, Hle, Nle, Nva, Moo

Met
Ile, Val, Hle, Nle, Nva, Moo

Nle
Ile, Val, Hle, Met, Nva, Moo

Asn
Asp, Glu, Gln, Cya, Thr

Pro
Aze, Pip, Hyp, Tfp, Cfp, Dmp, Tap, H3p, 4Ap, Cpp, Hym, Chy, Dfp

Gln
Asp, Asn, Glu, Cya, Thr, Hse

Arg
Arg, Dab, Dap, Har, Egd, Omr, Hci, Cit

Ser
Thr, Hse, allo-Threonine

Thr
Ser, Homothreonine, allo-Threonine

Val
Leu, Ile, Hle, Nva, Nle

Trp
Hfe, Phg, Bhf, Thi, Bta, Bromophenylalanine, Iodophenylalanine,

Chlorophenylalanine, Methylphenylalanine, Nitrophenylalanine, Tyr, Trp,

Naphthylalanine, Trifluoromethylphenylalanine

Tyr
Hfe, Phg, Bhf, Thi, Bta, Bromophenylalanine, Iodophenylalanine,

Chlorophenylalanine, Methylphenylalanine, Nitrophenylalanine, Tyr, Trp,

Naphthylalanine, Trifluoromethylphenylalanine

Linear Peptides

A general linear peptide is typically written from the N- to C-terminal direction as shown below:

NT-Xaa1-Xaa2-Xaa3-Xaa4- . . . Xaan-CT;

Therein

- 1. Xaax is the abbreviation, descriptor or symbol for amino acids or building blocks at specific sequence position x as shown in Table 5,
- 2. NT is a N-terminal group, e.g. ‘H’ (Hydrogen for a free N-terminal amino group) or an abbreviation for a specific terminating carboxylic acid like ‘Ac’ for acetic acid or other chemical group or structural formula of chemical groups linked to the N-terminal amino acid code (Xaa1) via a hyphen and
- 3. CT is a C-terminal group which is typically ‘OH’ or ‘NH₂’ (as terminal carboxylic acid or amide) or an abbreviation for a specific terminating amine linked to the C-terminal amino acid code (Xaan) via a hyphen.

Branched Peptides with Side Chains Modified by Specific Building Blocks or Peptides

A general linear, branched peptide is written from the N- to C-terminal direction as shown below:

NT-Xaa1-Xaa2-Xaa3(NT-Xab1-Xab2- . . . Xabn)- . . . Xaan-CT

Therein the statements 1.-3. of the description of linear peptides for the specification of Xaax, NT and CT in the main chain of the branched peptide apply.

The position of a branch is specified by parentheses after a Xaax abbreviation. Branches typically occur at lysine (Lys) residues (or similar), which means that the branch is attached to side chain ε-amino function of the lysine via an amide bond.

The content of the parenthesis describes the sequence/structure of the peptide branch ‘NT-Xab1-Xab2- . . . Xabn’. Herein

- 1. Xabx is the abbreviation, descriptor or symbol for amino acids or building blocks at specific sequence position x of the branch as shown in Table 3,
- 2. NT is a N-terminal group, e.g. an abbreviation for a specific terminating carboxylic acid like ‘Ac’ for acetic acid or other chemical group or structural formula of chemical groups linked to the N-terminal amino acid code (Xab1) via a hyphen and
- 3. the last building block of the branch Xabn, which connects the branch with the main chain by forming an amide bond with its own carboxyl function with the side chain amino function of this lysine (or similar residue).

Cyclic Peptides

An exemplaric general cyclic peptide written from the N- to C-terminal direction is shown below:

NT-Xaa1-[Xaa2-Xaa3-Xaa4- . . . Xaan]-CT;

Therein the statements 1.-3. of the description of linear peptides for the specification of Xaax, NT and CT in the main chain of the cyclic peptide apply. The characteristics of the peptide cycle are specified by square brackets.

- 1. The opening square bracket indicates the building block at whose side chain the cycle is initiated (cycle initiation residue) and
- 2. the closing square bracket indicates the building block at whose side chain the cycle is terminated (cycle termination residue).

The chemical nature of the connection between these two residues is

- 1. an amide bond in case that among those indicated residues one residue contains an amino function its side chain (e.g. Lys) while the other contains a carboxyl function in its side chain (e.g. Glu) or
- 2. a disulphide bond in case that those indicated residues/amino acids contain sulfhydryl moieties (e.g. Cys).

Cyclic Peptides Containing an Additional Cyclization Element (Yc)

A general extended cyclic peptide written from the N- to C-terminal direction is shown below:

NT-Xaa1-[Xaa2(Yc)-Xaa3-Xaa4- . . . Xaan]-CT;

Therein the statements 1.-3. of the description of linear peptides for the specification of Xaax, NT and CT in the main chain of the cyclic peptide apply. In addition, Yc is the cyclization element. As in case of cyclic peptides the characteristics of the cycle are specified by square brackets which indicate cycle initiation residue and cycle termination residue.

The content of the parentheses adjacent to the cycle initiation residue specifies the cyclization element Yc within the extended peptide cycle. The Yc element is linked to the side chain of said residue. Furthermore, the Yc element is linked to the side chain of the cycle termination residue. The chemical nature of the linkages between either of these residues the Yc element depend on side chain functionality of the corresponding amino acids Xaan. The linkage is a thioether if the side chain of Xaan contains a sulfhydryl group (e.g., Cys).

As non-limiting example the structure of Ac-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-OH is depicted below.

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Therein

- 1. Ac corresponds to NT in the general formula.
- 2. Cys, Pro, Pro, Thr, Gln, Phe and Cys correspond to Xaa1 to Xaa7 in the general formula.
- 3. OH corresponds to CT in the general formula.
- 4. The opening square bracket (‘[’) adjacent to the N-terminal cysteine in the sequence indicates that at this residue the cycle is initiated (cycle initiation residue).
- 5. The closing square bracket (‘]’) adjacent to the N-terminal cysteine in the sequence indicates that at this residue the cycle is terminated (cycle termination residue).
- 6. tMeBn within the parentheses adjacent to the Cys indicated as initiation residue specifies the cyclization element Yc. It is further bound to the Cys indicated as cycle termination residue. The Yc element is connected to said residues via thioether linkages.
- 7. To the remaining connection point of the tMeBn residue a DOTA chelator is attacted via a PP linker. For clarity terms like “Cys(tMeBn(DOTA-PP))” are included in the list of chemical structures in table 2

In an embodiment of the present invention, an amino acid or a peptide is attached to Xaa7, wherein a majority of the amino acids of this peptide are charged or polar and the net charge of the peptide is −2, −1, 0, +1 or +2.

For calculation of peptide net charges negatively charged amino acids are amino acids which bear acidic groups like —COOH or —SO₃H in their side chain and their net charge corresponds to the number of acidic groups, e.g. Asp or Glu with net charge −1.

For this calculation positively charged amino acids are amino acids which bear basic groups like amino or -guanidino in their side chain and their net charge corresponds to the number of basic groups, e.g. Lys or Arg with net charge+1.

Polar amino acids are amino acids which bear polar groups in their side chain. The polar groups are such as CONH2, OH, F, Cl, CN, and heterocycles like for instance imidazole in histidine.

The polar amino acids have a net charge of 0. For some nitrogen containing heterocycles the net charge is considered as 0 for our calculation although it is acknowledged that depending on the pH of the environment it might be protonated in an equilibrium and therefore positively charged to a certain extent.

The majority (50% or more) of the amino acids of this peptide are charged or polar.

Preferably the positive or negative charges are occasionally separated by a polar or non-polar amino acid.

In some embodiments the presence of negative charged amino acid is preferred at Xaa10.

In some embodiments the presence of positively charged amino acid is preferred at Xaa13, preferably Arg and arg.

In accordance with the present invention, the compound of the present invention may comprise a Z group. The Z group comprises a chelator and optionally a linker. As preferably used, a linker is an element, moiety, or structure which separates two parts of a molecule. In the present invention, the linker group forms covalent bonds with both the chelator group and the respective part of the compounds of invention where Z is attached. The linker group may, in principle, be any chemical group which is capable of forming bonds with both the chelator group and the part of the compounds of invention at the specified positions.

An important property or feature of a linker is that it spaces apart the chelator and the cyclic peptide part of the compound of invention. This is especially important in cases where the target binding ability of the cyclic peptide is compromised by the close proximity of the chelator. However, the overall linker length in its most extended conformer should not exceed 200 Å, preferably not more than 150 Å and most preferably not more than 100 Å.

In a preferred embodiment, the linker is —[X]_a—, wherein a is an integer from 1 to 10, and each X is an individual building block which is connected independently to its neighbors in the sequence by a functional group selected from comprising an amide linkage, a urea linkage, a carbamate linkage, an ester linkage, an ether linkage, a thioether linkage, a sulfonamide, a triazole and a disulfide linkage.

X₁is connected to the chelator- and, if present to X₂or to the compounds of invention at the specified positions. X_ais connected, if present to X_a-1and to the compounds of invention at the specified positions.

A more preferred class of linker groups is represented by is —[X]_a—, wherein a is an integer from 1 to 10, preferably, a is an integer from 1 to 8, 1 to 6, 1 to 5, 1 to 4 or 1 to 3, and each X is an individual building block which is connected independently to its neighbors in the sequence by a functional group selected from a group comprising an amide linkage, a urea linkage, a carbamate linkage, an ester linkage, an ether linkage, a thioether linkage, a sulfonamide linkage, a triazole linkage and a disulfide linkage.

In an embodiment the building block X is of general formula (8)

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wherein,
- fragment Lin², if present, and fragment Lin³, if present, are each individually and independently selected from the group comprising —CO—, —NR¹⁰—, —S—, —CO—NR¹⁰—, —CS—NR¹⁰—, —O—, -succinimide- and —CH₂—CO—NR¹⁰—; under the proviso that at least one of Lin²or Lin³is linked to R⁹with a carbon atom and the nitrogen atom of all nitrogen containing fragments is linked to R⁹;

wherein R¹⁰is selected from the group consisting of hydrogen and (C₁-C₄)alkyl;

and wherein R⁹is selected from —(C₁-C₁₀)alkylidene-, —(C₃-C₈)carbocyclo-, -arylene-, —(C₁-C₁₀)alkylidene-arylene-, -arylene-(C₁-C₁₀)alkylidene-, —(C₁-C₁₀)alkylidene-arylene-(C₁-C₁₀)alkylidene-, —(C₁-C₁₀)alkylidene-(C₃-C₈)carbocyclo-, —(C₃-C₈)carbocyclo-(C₁-C₁₀)alkylidene-, —(C₁-C₁₀)alkylidene-(C₃-C₈)carbocyclo-(C₁-C₁₀)alkylidene-, —(C₃-C₈)heterocyclo-, (C₁-C₁₀)alkylidene-(C₃-C₈)heterocyclo-, —(C₃-C₈)heterocyclo-(C₁-C₁₀)alkylidene-, —(C₁-C₁₀)alkylidene-(C₃-C₈)heterocyclo-(C₁-C₁₀)alkylidene-, —(CH₂CH₂O)_r—, and —(CH₂)_s—(CH₂CH₂O)_r—(CH₂)_t—;

and wherein

r is any integer from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;

s is any integer from 0, 1, 2, 3 and 4; and

t is any integer from 0, 1, 2, 3 and 4.

Preferably, apart from the linkage between X₁and the chelator, the linkage is an amide linkage. More preferably building block X₂to X_aare independently selected from the group of comprising an amino acid, a dicarboxylic acid and a diamine and the respective linkages are amides.

In an embodiment the building block X₂to X_ais preferably an amino acid, wherein the amino acid is selected from the group comprising conventional and unconventional amino acids. In an embodiment an amino acid is one selected from the group comprising β-amino acids, γ-amino acids, δ-amino acids, ε-amino acids and ω-amino acids. In a further embodiment an amino acid is a cyclic amino acid or a linear amino acid. It will be appreciated by a person skilled in the art that in case of an amino acid with stereogenic centers all stereoisomeric forms may be used in the building block X.

In an embodiment the building block X₂to X_ais preferably an amino acid, wherein the amino acid is selected from a group comprising amino acids which differ as to the spacing of the amino group from the carboxylic group. This kind of amino acid can be generically represented as follows:

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It is within the present invention that such amino acid is not further substituted. It is, however, also within the present invention that such amino acid is further substituted; preferably such substitution is CO—NH₂and/or Ac—NH—.

Representative of this kind of amino acid (structure 32) which can be used as a building block X are glycine (Gly), β-alanine (Bal), γ-aminobutyric acid (GABA), aminopentanoic acid, aminohexanoic acid and homologs with up to 10 CH₂groups.

Representative of this kind of amino acid (structure 33) which are more preferably used as a building block X are β-aminomethyl-benzoic acid, γ-aminomethyl-benzoic acid, anthranilic acid, 3-amino benzoic acid and 4-amino benzoic acid.

Relevant building blocks are diamines which are derived from amino acids (structure 32+33) by replacing NH₂with COOH, which are preferably used as a building block X are diamino ethane, 1,3-diamino propane, 1,4-diamino butane, 1,5-diamino pentane, 3-aminomethyl-aniline, 4-aminomethyl-aniline, 1,2-diamino benzene, 1,3-diamino benzene and 1,4-diamino benzene.

Relevant building blocks are dicarboxylic acids which are derived from amino acids (structure 32+33) by replacing COOH with NH₂, which are more preferably used as a building block X are malonic acid, succinic acid, glutaric acid, adipic acid, phthalic acid, terephthalic acid, isophthalic acid and 2, 3 or 4 carboxy-phenyl acetic acid.

In a further embodiment, the amino acid is an amino acid which contains, preferably as a backbone, a polyether. Preferably such polyether is polyethylene glycol and consists of up to 30 monomer units. Preferably, an amino acid comprising such polyether shows an increase in hydrophilicity compared to an amino acid not comprising such polyether. If incorporated into a building block X and, ultimately, into a linker group [X]_a, the result is typically an increase in hydrophilicity. A preferred embodiment of this kind of amino acid is depicted in the following, wherein it will be acknowledged that such amino acid may comprise 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 ethylene oxide moieties:

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Preferred ethylene glycol containing amino acids are Ttds (N-(3-{2-[2-(3-Amino-propoxy)-ethoxy]-ethoxy}-propyl)-succinamic acid) and O2Oc ([2-(2-Amino-ethoxy)-ethoxy]-acetic acid) the formula of which is as follows:

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In preferred embodiments, the linker comprises an oligomer or a monomer of only one specific amino acid selected from the group of Ttds, O2Oc, Apac, Gly, Bal, Gab, Mamb, Pamb, Ppac, 4Amc, Inp, Sni, Rni, Nmg, Cmp, PEG6, PEG12, PEG-amino acids and more preferably the linker is monomeric.

In another preferred embodiment, the linker comprises one building block X₂selected from the group of Ttds, O2Oc, Apac, Gly, Bal, Gab, Mamb Pamb, PEG6, PEG12 and PEG-amino acids and a second building block X₁which is directly bound to the amino-nitrogen of X₂and is directly attached to a chelator by a linkage selected from the group consisting of an amide linkage, a urea linkage, a carbamate linkage, an ester linkage, an ether linkage, a thioether linkage, a sulfonamide, a triazole and a disulfide linkage. X₁serves in this case as adapter to mediate the linkage of the different kind of attachment functionalities provided by a chelator to the nitrogen-atom of the amino acid X₂in the sense that X₁provides relevant complementary functionalities for the linkage of the chelator.

However, the use of linkers usually follows a purpose. In some circumstances it is necessary to space a larger moiety apart from a bioactive molecule in order to retain high bioactivity. In other circumstances introduction of a linker opens the chance to tune physicochemical properties of the molecule by introduction of polarity or multiple charges. In certain circumstances it might be a strength and achievement if one can combine the chelator with a bioactive compound without the need for such linkers. Especially in those compounds of the present invention where the chelator is attached to Yc of formula (X) linking the S atom of Xaa1 and the S atom of Xaa7 under the formation of two thioether linkages typically perform excellently without the use of any dedicated linkers.

In an embodiment, the compound of the invention comprises a chelator. Preferably, the chelator is part of the compound of the invention, whereby the chelator is either directly or indirectly such as by a linker attached to the compound of the invention. A preferred chelator is a chelator which forms metal chelates preferably comprising at least one radioactive metal. The at least one radioactive metal is preferably useful in or suitable for diagnostic and/or therapeutic and/or theraognostic use and is more preferably useful in or suitable for imaging and/or radiotherapy.

Chelators in principle useful in and/or suitable for the practicing of the instant invention including diagnosis and/or therapy of a disease are known to the person skilled in the art. A wide variety of respective chelators is available and has been reviewed, e.g. by Banerjee et al. (Banerjee, et al., Dalton Trans, 2005, 24: 3886), and references therein (Price, et al., Chem Soc Rev, 2014, 43: 260; Wadas, et al., Chem Rev, 2010, 110: 2858). Such chelators include, but are not limited to linear, cyclic, macrocyclic, tetrapyridine, N3S, N2S2 and N4 chelators as disclosed in U.S. Pat. No. 5,367,080 Å, U.S. Pat. No. 5,364,613 Å, U.S. Pat. No. 5,021,556 Å, U.S. Pat. No. 5,075,099 Å and U.S. Pat. No. 5,886,142 Å.

Representative chelating agents, also referred to herein as chelators, and their derivatives suitable in the practicing of the present invention include, but are not limited to 99mTc(CO)3-chelators, AAZTA, BAT, CDTA, DTA, DTPA, CY-DTA, DTCBP, CHX-A″-DTPA, CTA, cyclam, cyclen, TETA, Sarcophagine, CPTA, TEAMA, Crown, Cyclen, DO3A, DO2A, TRITA, DATA, DFO, DATA(M), DATA(P), DATA(Ph), DATA(PPh), DEDPA, H4octapa, H2dedpa, HSdecapa, H2azapa, H2CHX DEDPA, DFO-Chx-MAL, DFO-p-SCN, DFO-1AC, DFO-BAC, p-SCN-Bn-DFO, DFO-pPhe-NCS, DFO-HOPO, DFC, Diphosphine, DOTA, DOTAGA, DOTA-MFCO, DOTAM-mono-acid, nitro-DOTA, nitro-PA-DOTA, p-NCS-Bz-DOTA, PA-DOTA, DOTA-NCS, DOTA-NHS, CB-DO2A, PCTA, p-NH2-Bn-PCTA, p-SCN-Bn-PCTA, p-SCN-Bn-DOTA, DOTMA, NB-DOTA, H4NB-DOTA, H4TCE-DOTA, 3,4,3-(Li-1,2-HOPO), TREN(Me-3,2-HOPO), TCE-DOTA, DOTP, DOXP, p-NCS-DOTA, p-NCS-TRITA, TRITA, TETA, 3p-C-DEPA, 3p-C-DEPA-NCS, p-NH2-BN-OXO-DO3A, p-SCN-BN-TCMC, TCMC, 4-Aminobutyl-DOTA, Azido-mono-amide-DOTA, BCN-DOTA, Butyne-DOTA, BCN-DOTA-GA, DOA3P, DO2a2p, DO2A(trans-H2do2a), DO3A, DO3A-Thiol, DO3AtBu-N-(2-aminoethyl)ethanamide, DO2AP, CB-DO2A, C3B-DO2A, HP-DO3A, DOTA-NHS-ester, Maleimide-DOTA-GA, Maleimido-mono-aminde-DOTA, Maleimide-DOTA, NH2-DOTA-GA, NH2-PEG4-DOTA-GA, p-NH2-Bn-DOTA, p-N02-Bn-DOTA, p-SCN-Bn-DOTA, p-SCN-Bz-DOTA, TA-DOTA, TA-DOTA-GA, OTTA, DOXP, TSC, DTC, DTCBP, PTSM, ATSM, FSC, H2ATSM, H2PTSM, Dp44mT, DpC, Bp44mT, QT, hybrid thiosemicarbazone-benzothiazole, thiosemicarbazone-styrylpyridine tetradentate ligands H2L2-4, HBED, HBED-CC, dmHBED, dmEHPG, HBED-nn, SHBED, Br-Me2HBED, BPCA, HEHA, BF-HEHA, Deferiprone, THP, HOPO, HYNIC (2-hydrazino nicotinamide), NHS-HYNIC, HYNIC-Kp-DPPB, HYNIC-Ko-DPPB, (HYNIC)(tricine)2, (HYNIC)(EDDA)Cl, p-EDDHA, AIM, AIM A, IAM B, MAMA, MAMA-DGal, MAMA-MGal, MAMA-DA, MAMA-HAD, Macropa, Macropaquin, Macroquin-5O3, NxS4-x, N2S2, N3S, N4, MAG3B, NOTA, NODAGA, SCN-Bz-NOTA-R, NOT-P (NOTMP), NOTAM, p-NCS-NOTA, TACN, TACN-TM, NETA, NETA-monoamine, p SCN-PhPr-NE3TA, C-NE3TA-NCS, C-NETA-NCS, 3p-C-NETA, NODASA, NOPO, NODA, NO2A, N-Benzyl-NODA, NODA-MPAA, C-NOTA, BCNOT-Monoamine, Maleimido-mono-amide-NOTA, NO2A-Azide, NO2A-Butyne, NO2AP, NO3AP, N-NOTA, Oxo-DO3A, p-NH2-Bn-NOTA, p-NH₂-Bn-oxo-DO3A, p-NO2-Bn-Cyclen, PSC, p-SCN-Bn-NOTA, NOTP, p-SCN-Bn-oxo-DO3A, TRAP, PEPA, BF-PEPA, Pycup, Pycup2A, pycup1A1Bn, pycup2Bn, RESCA, SarAr-R, Diamsar, AmBaSar-R, siamSar, Sar, Tachpyr, tachpyr-(6-Me), TAM A, TAM B, TAME, TAME-Hex, THP-Ph-NCS, THP-NCS, THP-TATE, NTP, H3THP, THPN, CB-TE2A, PCB-TE1A1P, TETA-NHS, CPTA, CPTA-NHS, CB-TE1K1P, CB-TE2A, TE2A, H2CB-TE2A, TE2P, CB-TE2P, MM-TE2A, DM-TE2A, 2C-TETA, 6C-TETA, BAT, BAT-6, NHS-BAT ester, SSBAT, SCN-CHX-A-DTPA-P, SCN-TETA, TMT-amine, p-BZ-HTCPP.

HYNIC, DTPA, EDTA, DOTA, TETA, bisamino bisthiol (BAT) based chelators as disclosed in U.S. Pat. No. 5,720,934; Desferrioxamin (DFO) as disclosed in (Doulias, et al., Free Radic Biol Med, 2003, 35: 719), tetrapyridine and N₃S, N₂S₂and N₄chelators as disclosed in U.S. Pat. No. 5,367,080 Å, U.S. Pat. No. 5,364,613 Å, U.S. Pat. No. 5,021,556 Å, U.S. Pat. No. 5,075,099 Å, U.S. Pat. No. 5,886,142 Å, whereby all of the references are included herein by reference in their entirety. 6-Amino-6-methylperhydro-1,4-diazepine-N,N′,N″,N″-tetraacetic acid (AAZTA) is disclosed in Pfister et al., (Pfister, et al., EJNMMI Res, 2015, 5: 74), Deferiprone, a 1,2-dimethyl-3,4-hydroxypyridinone and Hexadentate tris(3,4-hydroxypyridinone) THP) are disclosed in Cusnir et al. (Cusnir, et al., Int J Mol Sci, 2017, 18), monoamine-monoamide dithiol (MAMA)-based chelators are disclosed in Demoin et al. (Demoin, et al., Nucl Med Biol, 2016, 43: 802), MACROPA and analogues are disclosed in Thiele et al. (Thiele, et al., Angew Chem Int Ed Engl, 2017, 56: 14712), 1,4,7,10,13,16-hexaazacyclohexadecane-N,N′,N″,N′″,N″″N′″″-hexaacetic acid (HEHA) and PEPA analogues are disclosed in Price and Orvig (Price, et al., Chem Soc Rev, 2014, 43: 260), Pycup and analogous are disclosed in Boros et al. (Boros, et al., Mol Pharm, 2014, 11: 617), N, N-bis(2-hydroxybenzyl)ethylenediamine-N,N-diacetic acid (HBED), 1,4,7,10-tetrakis (carbamoylmethyl)-1,4,7,10-tetraazacyclododecane (TCM), 2-[(carboxymethyl)]-[5-(4-nitrophenyl-1-[4,7,10-tris-(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]pentan-2-yl)-amino]acetic acid (3p-C-DEPA), CB-TE2A, TE2A, TE1A1P, Diamsar, 1-N-(4-Aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]-eicosane-1,8-diamine (SarAr), NETA, N,N0,N00, tris(2-mercaptoethyl)-1,4,7-triazacyclononane (TACN-TM), {4-[2-(Bis-carboxymethyl-amino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid (NETA), diethylenetriaminepentaacetic acid (DTP), 3-({4,7-Bis-[(2-carboxy-ethyl)-hydroxy-phosphinoylmethyl]-[1,4,7]triazonan-1-ylmethyl}-hydroxy-phosphinoyl)-propionic acid (TRAP), NOPO, H4octapa, SHBED, BPCA, 3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-1(15),11,13-triene-3,6,9,-triacetic acid (PCTA), and 1,4,7,10,13-pentaazacyclopentadecane-N,N′,N″,N′″,N′″-pentaacetic acid (PEPA) are disclosed in Price and Orvig (Price, et al., Chem Soc Rev, 2014, 43: 260), 1-hydroxy-2-pyridone ligand (HOPO) is disclosed in Allott et al. (Allott, et al., Chem Commun (Camb), 2017, 53: 8529), [4-Carboxymethyl-6-(carboxymethyl-methyl-amino)-6-methyl-[1,4]diazepan-1-yl]-acetic acid (DATA) is disclosed in Tornesello et al. (Tomesello, et al., Molecules, 2017, 22: 1282), tetrakis(aminomethyl)methane (TAM) and analogues are disclosed in McAuley 1988 (McAuley, et al., Canadian Journal of Chemistry, 1989, 67: 1657), Hexadentate tris(3,4-hydroxypyridinone) (THP) and analogues are disclosed in Ma et al. (Ma, et al., Dalton Trans, 2015, 44: 4884).

The diagnostic and/or therapeutic use of some of the above chelators is described in the prior art. For example, 2-hydrazino nicotinamide (HYNIC) has been widely used in the presence of a coligand for incorporation of ^99mTc and ^186,188Re (Schwartz, et al., Bioconjug Chem, 1991, 2: 333; Babich, et al., J Nucl Med, 1993, 34: 1964; Babich, et al., Nucl Med Biol, 1995, 22: 25); DTPA is used in Octreoscan® for complexing ¹¹¹In and several modifications are described in the literature (Li, et al., Nucl Med Biol, 2001, 28: 145; Brechbiel, et al., Bioconjug Chem, 1991, 2: 187); DOTA type chelators for radiotherapy applications are described by Tweedle et al. (U.S. Pat. No. 4,885,363); other polyaza macrocycles for chelating trivalent isotopes metals are described by Eisenwiener et al. (Eisenwiener, et al., Bioconjug Chem, 2002, 13: 530); and N₄-chelators such as a ^99mTc-N₄-chelator have been used for peptide labeling in the case of minigastrin for targeting CCK-2 receptors (Nock, et al., J Nucl Med, 2005, 46: 1727).

In an embodiment the metal chelator is selected from the group, but not limited to, comprising DOTA, DOTAGA, NOTA, NODAGA, NODA-MPAA, HBED, TETA, CB-TE2A, DTPA, DFO, Macropa, HOPO, TRAP, THP, DATA, NOTP, sarcophagine, FSC, NETA, H4octapa, Pycup, N_xS_4-x(N4, N2S2, N3S), Hynic, ^99mTc(CO)3-Chelators and their analogs, wherein

- DOTA stands for 1,4,7,10-tetrazacyclododecane-1,4,7,10-tetraacetic acid,
- DOTAGA stand for 1,4,7,10-tetraazacyclodocecane,1-(glutaric acid)-4,7,10-triacetic acid,
- NOTA stands for 1,4,7-triazacyclononanetriacetic acid,
- NODAGA stands for 1,4,7-triazacyclononane-N-glutaric acid-N′,N″-diacetic acid,
- NODA-MPAA stands for 1,4,7-triazacyclononane-1,4-diacetate-methyl phenylacetic acid,
- HBED stands for bis(2-hydroxybenzyl) ethylenediaminediacetic acid,
- TETA stands for 1,4,8,11-tetraazacyclododecane-1,4,8,11-tetraacetic acid,
- CB-TE2A stands for 4,11-bis-(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]-hexadecane,
- DTPA stands for diethylenetriaminepentaacetic acid,
- DFO stands for the Desferal or Desferrioxamine type group of chelators, the chemical name of the non-limiting example is N-[5-({3-[5-(Acetyl-hydroxy-amino)-pentylcarbamoyl]-propionyl}-hydroxy-amino)-pentyl]-N′-(5-amino-pentyl)-N′-hydroxy-succinamide,
- Macropa stands for N,N′-bis[(6-carboxy-2-pyridyl)methyl]-4,13-diaza-18-crown,
- HOPO stands for the octadentate hydroxypyridinone type group of chelators, the structure of a non-limiting example is shown below,
- TRAP stands for 3-({4,7-Bis-[(2-carboxy-ethyl)-hydroxy-phosphinoylmethyl]-[1,4,7]triazonan-1-ylmethyl}-hydroxy-phosphinoyl)-propionic acid,
- THP stands for Hexadentate tris(3,4-hydroxypyridinone),
- DATA stands for [4-Carboxymethyl-6-(carboxymethyl-methyl-amino)-6-methyl-[1,4]diazepan-1-yl]-acetic acid
- NOTP stands for 1,4,7-triazacyclononane-N,N′N″-tris(methylene phosphonic) acid),
- Sarcophagine stands for 3,6,10,13,16,19-hexaazabicyclo[6.6.6]icosane,
- FSC stands for 3,15,27-Triamino-7,19,31-trihydroxy-10,22,34-trimethyl-1,13,25-trioxa-7,19,31-triaza-cyclohexatriaconta-9,21,33-triene-2,8,14,20,26,32-hexaone,
- NETA, {4-[2-(Bis-carboxymethyl-amino)-ethyl]-7-carboxymethyl-[1,4,7]triazonan-1-yl}-acetic acid
- H4octapa, N,N′-(6-carboxy-2-pyridylmethyl)-N,N′-diacetic acid-1,2-diaminoethane
- Pycup stands for 1,8-(2,6-Pyridinedimethylene)-1,4,8,11-tetraazacyclo-tetradecane,
- N_xS_4-x(N4, N2S2, N3S) stands for a group of tetradentate chelators with N-atoms (basic amine or non-basic amide) and thiols as donors stabilizing Tc-complexes, especially Tc(V)-oxo complexes. The structure of one representative non-limiting example MAG3 is shown below, and
- MAG3 stands for {2-[2-(3-Mercapto-propionylamino)-acetylamino]-acetylamino}-acetic acid,
- HYNIC stands for 6-Hydrazino-nicotinic acid,
- ^99mTc(CO)₃-Chelators stands for bi- or tridendate chelators capable of forming stable complexes with technetium tricarbonyl fragments,
- and with the chemical structures thereof being as follows:

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In a preferred embodiment, the metal chelator is selected from the group consisting of DOTA, DOTAGA, NOTA, NODAGA, NODA-MPAA, HBED, CB-TE2A, DFO, THP, N4 and analogs thereof.

In a more preferred embodiment, the metal chelator is selected from the group consisting of DOTA, DOTAGA, NOTA, N4Ac and NODAGA and their analogs thereof.

It will be acknowledged by the persons skilled in the art that the chelator, in principle, may be used regardless whether the compound of the invention is used in or suitable for diagnosis or therapy. Such principle is, among others, outlined in international patent application WO 2009/109332 Å1.

It will be further acknowledged by the persons skilled in the art that the presence of a chelator in the compound of the invention includes, if not stated otherwise, the possibility that the chelator is complexed to any metal complex partner, i.e. any metal which, in principle, can be complexed by the chelator. An explicitly mentioned chelator of a compound of the invention or the general term chelator in connection with the compound of the invention refers either to the uncomplexed chelator as such or to the chelator to which any metal complex partner is bound, wherein the metal complex partner is any radioactive or non-radioactive metal complex partner. Preferably the chelator metal complex, i.e. the chelator to which the metal complex partner is bound, is a stable chelator metal complex.

Non-radioactive chelator metal complexes have several applications, e.g. for assessing properties like stability or activity which are otherwise difficult to determine. One aspect is that cold variants of the radioactive versions of the metal complex partner (e.g. non-radioactive Gallium, Lutetium or Indium complexes as described in the examples) can act as surrogates of the radioactive compounds. Furthermore, they are valuable tools for identifying metabolites in vitro or in vivo, as well as for assessing toxicity properties of the compounds of invention.

Additionally, chelator metal complexes can be used in binding assays utilizing the fluorescence properties of some metal complexes with distinct ligands (e.g. Europium salts).

Chelators can be synthesized or are commercially available with a wide variety of (possibly already activated) groups for the conjugation to peptides or amino acids. Direct conjugation of a chelator to an amino-nitrogen of the respective compound of invention is well possible for chelators selected from the group consisting of DOTA, DOTAGA, NOTA, NODAGA, NODA-MPAA, HBED, TETA, CB-TE2A, DTPA, DFO, DATA, sarcophagine, N4, MAG3 and Hynic, preferably DOTA, DOTAGA, NOTA, NODAGA, NODA-MPAA, CB-TE2A, and N4. The preferred linkage in this respect is an amide linkage.

Functional groups at a chelator which are ideal precursors for the direct conjugation of a chelator to an amino-nitrogen are known to the person skilled in the art and include but are not limited to carboxylic acid, activated carboxylic acid, e.g. active ester like for instance NHS-ester, pentafluorophenol-ester, HOBt-ester and HOAt-ester, isothiocyanate.

Functional groups at a chelator which are ideal precursors for the direct conjugation of a chelator to a carboxylic group of a peptide are known to the person skilled in the art and include but are not limited to alkylamino and arylamino nitrogens. Respective chelator reagents are for commercially available some chelators, e.g. for DOTA with either alkylamino or arylamino nitrogen.

It will be acknowledged by a person skilled in the art that the radioactive nuclide which is or which is to be attached to the compound of the invention, is selected taking into consideration the disease to be treated and/or the disease to be diagnosed, respectively, and/or the particularities of the patient and patient group, respectively, to be treated and to be diagnosed, respectively.

In an embodiment of the present invention, the radioactive nuclide is also referred to as radionuclide. Radioactive decay is the process by which an atomic nucleus of an unstable atom loses energy by emitting ionizing particles (ionizing radiation). There are different types of radioactive decay. A decay, or loss of energy, results when an atom with one type of nucleus, called the parent radionuclide, transforms to an atom with a nucleus in a different state, or to a different nucleus containing different numbers of protons and neutrons. Either of these products is named the daughter nuclide. In some decays the parent and daughter are different chemical elements, and thus the decay process results in nuclear transmutation (creation of an atom of a new element). For example, the radioactive decay can be alpha decay, beta decay, and gamma decay. Alpha decay occurs when the nucleus ejects an alpha particle (helium nucleus). This is the most common process of emitting nucleons, but in rarer types of decays, nuclei can eject protons, or specific nuclei of other elements (in the process called cluster decay). Beta decay occurs when the nucleus emits an electron (β⁻-decay) or positron (β⁺-decay) and a type of neutrino, in a process that changes a proton to a neutron or the other way around. By contrast, there exist radioactive decay processes that do not result in transmutation. The energy of an excited nucleus may be emitted as a gamma ray in gamma decay, or used to eject an orbital electron by interaction with the excited nucleus in a process called internal conversion, or used to absorb an inner atomic electron from the electron shell whereby the change of a nuclear proton to neutron causes the emission of an electron neutrino in a process called electron capture (EC), or may be emitted without changing its number of proton and neutrons in a process called isomeric transition (IT). Another form of radioactive decay, the spontaneous fission (SF), is found only in very heavy chemical elements resulting in a spontaneous breakdown into smaller nuclei and a few isolated nuclear particles.

In a preferred embodiment of the present invention, the radionuclide can be used for labeling of the compound of the invention.

In an embodiment of the present invention, the radionuclide is suitable for complexing with a chelator, leading to a radionuclide chelate complex.

In a further embodiment one or more atoms of the compound of the invention are of non-natural isotopic composition, preferably these atoms are radionuclides; more preferably radionuclides of carbon, oxygen, nitrogen, sulfur, phosphorus and halogens: These radioactive atoms are typically part of amino acids, in some case halogen containing amino acids, and/or building blocks and in some cases halogenated building blocks each of the compound of the invention.

In a preferred embodiment of the present invention, the radionuclide has a half-life that allows for diagnostic and/or therapeutic medical use. Specifically, the half-life is between 1 min and 100 days.

In a preferred embodiment of the present invention, the radionuclide has a decay energy that allows for diagnostic and/or therapeutic medical use. Specifically, for γ-emitting isotopes, the decay energy is between 0.004 and 10 MeV, preferably between 0.05 and 4 MeV, for diagnostic use. For positron-emitting isotopes, the decay energy is between 0.6 and 13.2 MeV, preferably between 1 and 6 MeV, for diagnostic use. For particle-emitting isotopes, the decay energy is between 0.039 and 10 MeV, preferably between 0.4 and 6.5 MeV, for therapeutic use.

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 the diagnostic and/or therapeutic medical use. Furthermore, the daughter nuclides are either stable or further decay in a way that does not interfere with or even support the diagnostic and/or therapeutic medical use. Representative radionuclides which may be used in connection with the present invention are summarized in Table 7.

TABLE 7

Key properties of relevant radionuclides-half life, decay types and decay energies

Half-
Half-
Half-

life
life
life

Energy
Additional decays

Radionuclide
(min)
(hours)
(days)
Decay
(MeV)
(energy [MeV])

Carbon

C-11
20.4
0.34

ECβ+
1.982

Nitrogen

N-13
9.97
0.17

ECβ+
2.220

Oxygen

O-15
2.00

ECβ+
2.754

Fluorine

F-18
110
1.83

β+
1.656

Mg-28

20.9

β−
1.832

Aluminum

Al-28
2.24
0.04

β−
4.642

Al-29
6.56

β−
3.690

Silicon

Si-31
157
2.62

β−
1.492

Phosphorus

P-30
2.50
0.04

β+
4.232

P-32

14.3
β−
1.170

P-33

25.4
β−
0.077

Sulphur

S-35

87.4
β−
0.167

S-37
5.00
0.08

S-38

2.80

β−
2.937

Chlorine

Cl-34m1
32.0
0.53

EC
5.693

Cl-38
37.2
0.62

β−
4.917

Cl-39
55.6
0.93

β−
3.422

Scandium

Sc-43

3.89

EC
2.221

Sc-44

3.97

β+
0.632

Sc-44m1

58.6
2.44
IT
0.271
98.8% IT (0.27086), 1.2%

EC (3.924)

Sc-46

83.8
β−
2.367

Sc-47

80.4
3.35
β−
0.601

Sc-48

43.7
1.82
β−
3.988

Sc-49
57.4
0.96

β−
2.002

Titanium

Ti-45
185
3.08

EC
2.062

Ti-51
5.76

β−
2.472

Vanadium

V-47
32.6
0.54

β+
2.931

V-48

16.2
EC
4.013

V-49

330
EC
0.602

V-52
3.74

β−
3.975

Chromium

Cr-48

23.0

EC
1.655

Cr-49
42.1
0.70

β+
2.628

Cr-51

27.7
EC
0.753

Cr-55
3.50

β−
2.603

Cr-56
5.94

β−
1.630

Manganese

Mn-51
46.2
0.77

β+
2.185

Mn-52m1
21.1
0.35

EC
5.091
98.25% EC (5.091), 1.75%

IT (0.3796)

Mn-52

5.59
β+
3.689

Mn-54

312
EC
1.377

Mn-56

2.58

β−
3.696

Iron

Fe-52

8.28

EC
2.375

Fe-53m1

2.54

IT
3.042

Fe-53

8.51

EC
3.742

Fe-59

44.5
β−
1.565

Fe-61

5.98

β−
3.977

Cobalt

Co-55

17.5

EC
3.451

Co-56

78.8
EC
4.567

Co-57

271
EC
0.836

Co-58m1

9.15

IT
0.026

Co-58

70.8
EC
2.308

Co-60m1
10.5
0.17

IT
0.059
99.76% IT (0.05932),

0.24% β− (2.882)

Co-61

1.65

β−
1.324

Co-62m1
13.9
0.23

β−
5.337

Nickel

Ni-56

146
6.10
EC
2.133

Ni-57

36.1
1.50
β+
3.262

Ni-63

β−
0.067

Ni-65

2.52

β−
2.138

Ni-66

54.6
2.28
β−
0.252

Copper

Cu-60
23.2
0.39

EC
6.128

Cu-61

3.41

EC
2.238

Cu-62
9.74
0.16

EC
3.959

Cu-64

12.7

β+
0.653
61.5% EC (1.674), 38.5%

β− (0.5797)

Cu-66
5.10
0.09

β−
2.641

Cu-67

2.58
β−
0.580

Cu-68m1
3.75

IT
0.722
84% IT (0.72163), 16%

β− (5.162)

Cu-69
2.85

β−
2.681

Zinc

Zn-60
2.38

EC
4.171

Zn-62

9.26

EC
1.620

Zn-63
38.1
0.64

EC
3.366

Zn-65

244
EC
1.352

Zn-69m1

13.8

IT
0.438
99.997% IT (0.43818),

0.003% β⁻ (1.348)

Zn-69
57.0
0.95

β−
0.910

Zn-71m1

3.92

β−
2.970
99.95% β− (2.97), 0.05%

IT (0.15986)

Zn-71
2.45

β−
2.810

Zn-72

46.5
1.94
β−
0.443

Gallium

Ga-65
15.2
0.25

EC
3.255

Ga-66

9.40

EC
5.175

Ga-67

78.2
3.26
EC
1.001

Ga-68
68.0
1.13

β+
2.921

Ga-70
21.1
0.35

β−
1.652
99.59% β− (1.652), 0.41%

EC (0.65456)

Ga-72

14.1

β−
3.998

Ga-73

4.91

β−
1.598

Ga-74
8.12
0.14

β−
5.373

Selenium

Se-70
41.0
0.68

β+
2.412

Se-72
504
8.40

EC
0.362

Se-73m
39.0
0.65

IT
2.761
27.4% EC (2.761), 72.6%

IT (0.03608)

Se-73
429
7.15

EC
2.725

Se-75

120
EC
0.865

Se-79m1
3.92

IT
0.096
99.94% IT (0.09622),

0.06% (0.247)

Se-81m1
57.2
0.95

IT
0.103
99.95% IT (0.10253),

0.05% β− (1.689)

Se-81
18.5
0.31

β−
1.587

Se-83
22.3
0.37

β−
3.673

Se-84
3.26

β−
1.836

Bromine

Br-73
3.40

EC
4.580

Br-74m1
41.5
0.69

EC
9.921

Br-74
25.3
0.42

EC
6.925

Br-75
98.0
1.63

EC
3.062

Br-76

16.2

β+
3.941

Br-77

57.0
2.38
β+
0.342

Br-78
6.64
0.11

EC
3.574
99.99% EC (3.574), 0.01%

β− (0.72746)

Br-80m1
265.20
4.42

IT
0.085

Br-80
17.40
0.29

EC
1.870
1.87 (EC), 2,004 (β−),

EC = 91.7, β− = 8.3

Br-82

35.30
1.47
β−
3.090

Br-83
143.40
2.39

β−
0.972

Br-84
31.80
0.53

β−
4.656

Br-84m1
6.00

β−
4.960

Br-85
2.90

β−
2.905

Yttrium

Y-83
7.08

EC
4.470

Y-83m1
2.85

EC
4.532
4.532 (ECβ+), 0.062 (IT),

ECβ+ = 60, IT = 40

Y-84

Y-84m1
39.50
0.66

EC
6.490

Y-85
160.80
2.68

EC
3.250

Y-85m1
291.60
4.86

EC
3.270

Y-86m1
48.00
0.80

IT
0.218

Y-86

14.74

ECβ+
4.22

Y-87m1

13.37

IT
0.381
0.381 (IT), 2.243 (ECβ+),

IT = 98.43, ECβ+ = 1.57

Y-87

80.30
3.35
ECβ+
1.862

Y-88

106.64
ECβ+
3.623

Y-90m1

3.19

IT
0.682

Y-90

64.08
2.67
β−
2.280

Y-91m1
49.71
0.83

IT

Y-91

58.51
β−

Y-92

3.54

β−
3.639

Y-93

10.10

β−
2.893

Y-94
19.10
0.32

β−
4.919

Y-95
10.70
0.18

β−
4.420

Zirconium

Zr-84
25.90

ECβ+

Zr-85
7.86

ECβ+
4.690

Zr-86

16.50

ECβ+
1.480

Zr-87
100.80
1.68

ECβ+
3.665

Zr-88

83.40
EC
0.670

Zr-89m1
4.18

IT
0.588
3.420 (ECβ+), 0.588 (IT),

ECβ+ = 6.23, IT = 93.77

Zr-89

78.43
3.27
β+
0.9

Zr-95

63.98
β−
1.125

Zr-97

16.90

β−
2.658

Niobium

Nb-87
2.60

ECβ+
5.170

Nb-87m1
3.70

ECβ+
5.170

Nb-88
14.50
0.24

ECβ+
7.200

Nb-88m1
7.80

ECβ+
7.200

Nb-89
114.00
1.90

ECβ+
4.290

Nb-89m1
70.80
1.18

ECβ+
4.290

Nb-90

14.60

ECβ+
6.111

Nb-91m1

60.86
IT
0.104
0.104 (IT), 1.357 (ECβ+),

IT = 93, ECβ+ = 7

Nb-95m1

86.60
3.61
IT
0.236

Nb-95

35.15
β−
0.926

Nb-96

23.35

β−
3.187

Nb-97
72.10
1.20

β−
1.934

Nb-98m1
51.50
0.86

β−
4.586

Molybdenum

Mo-88
8.00

ECβ+
3.720

Mo-89
2.04

ECβ+
5.580

Mo-90

5.67

ECβ+
2.489

Mo-91
15.49

ECβ+
4.434

Mo-93m1

6.85

IT.
2.830
IT = 99.88, ECβ+ = 0.12

ECβ+

Mo-99

66.00
2.75
β−
1.375

Mo-101
14.62
0.24

β−
2.824

Mo-102
11.30

β−
1.010

Technetium

Tc-91
3.14
0.05

ECβ+
6.220

Tc-91m1
3.30
0.06

ECβ+
6.570
6.57 (ECβ+), 0.35 (IT);

ECβ+ = 100, IT < 1

Tc-92
4.23
0.07

ECβ+
7.870

Tc-93m1
43.50
0.73

IT
0.392
3.593 (EC+), 0.392 (IT),

IT = 76.6, EC+ = 23.4

Tc-93

2.75

EC
3.201

Tc-94m1
52.00
0.87

β+
2.36
1.730 (ECβ+), 0.075 (IT);

ECβ+ ≈ 100, IT < 0.1

Tc-94

4.90

ECβ+
4.256

Tc-95m1

61.00
ECβ+
1.730
1.730 (ECβ+), 0.039 (IT);

ECβ+ = 96.12, IT = 3.88

Tc-95

20.00

EC
1.691

Tc-96m1
51.50
0.86

IT
0.034
3.007 (ECβ+), 0.034 (IT),

IT = 98.0, ECβ+ = 2.0

Tc-96

102.72
4.28
EC
2.973

Tc-97m1

87.00
IT
0.097

Tc-99m1

6.02

IT
0.143

Tc-101
14.20
0.24

β−
1.614

Tc-102m1
4.35

β−
4.530
4.53 (β−), 0.0 (IT), β− = 98,

IT = 2

Tc-104
18.20
0.30

β−
5.600

Tc-105
7.60
0.13

β−
3.640

Ruthenium

Ru-92
3.65

ECβ+
4.500

Ru-94
51.80
0.86

EC
1.593

Ru-95

1.64

ECβ+
2.572

Ru-97

69.60
2.90
EC
1.115

Ru-103

39.28
β−
0.763

Ru-105

4.44

β−
1.917

Ru-106

368.20
β−
0.039

Ru-107
3.76
0.06

β−
2.940

Ru-108
4.55
0.08

β−
1.360

Rhodium

Rh-95
5.02
0.08

ECβ+
5.110

Rh-95m1
1.96
0.03

IT
0.543
5.653 (ECβ+), 0.543 (IT);

% ECβ+ = 12, IT = 88

Rh-96
9.90
0.17

ECβ+
6.446

Rh-97
30.70
0.51

ECβ+
3.520

Rh-97m1
46.20
0.77

ECβ+
3.779
3.779 (ECβ+), 0.259 (IT);

ECβ+ = 94.4, IT = 5.6

Rh-98
8.70
0.15

ECβ+
5.057

Rh-98m1
3.50
0.06

ECβ+
5.057
5.057 (ECβ+), 0.0 (IT);

ECβ+ > 0

Rh-99m1

4.70

ECβ+
2.167
2.167 (ECβ+), 0.064 (IT),

ECβ+ > 99.84, IT < 0.16

Rh-99

16.00
ECβ+
2.130

Rh-100

20.80

ECβ+
3.630

Rh-101m1

104.16
4.34
EC
0.699
0.699 (EC), 0.157 (IT),

EC = 92.8, IT = 7.2

Rh-102

207.00
ECβ+
2.323
2.323 (ECβ+), 1.150(β−),

ECβ+ = 80, β− = 20

Rh-103m1
56.12
0.94

IT
0.040

Rh-104m1
4.34

IT
0.129
0.129 (IT), 2.570(β−), IT =

99.87, β⁻ = 0.13

Rh-105

35.36
1.47
β−
0.567

Rh-106m1
132.00
2.20

β−
3.678

Rh-107
21.70
0.36

β−
1.511

Rh-108m1
6.00

β−
4.510

Palladium

Pd-97
3.10

ECβ+
4.800

Pd-98
17.70

ECβ+
1.873

Pd-99
21.40

ECβ+
3.365

Pd-100

87.12
3.63
EC
0.361

Pd-101

8.27

ECβ+
1.980

Pd-103

16.96
EC
0.543

Pd-109

13.43

β−
1.116

Pd-109m1
4.70

IT
0.189

Pd-111
23.40
0.39

β−
2.190

Pd-111m1

5.50

IT
0.172
0.172 (IT), 2.362 (β⁻); IT =

73, β− = 27

Pd-112

21.03

β−
0.288

Pd-114
2.42
0.04

β−
1.451

Silver

Ag-100
2.01

ECβ+
7.050

Ag-100m1
2.24

ECβ+
7.066
7.066 (ECβ⁺), 0.015 (IT)

Ag-101
11.10

ECβ+
4.200

Ag-102
12.90
0.22

ECβ+
5.920

Ag-102m1
7.70

ECβ+
5.929
5.929 (ECβ+), 0.009 (IT),

ECβ+ = 51, IT = 49

Ag-103
65.70
1.10

ECβ+
2.688

Ag-104m1
33.50
0.56

ECβ+
4.286
4.286 (ECβ+), 0.007 (IT),

ECβ+ ≈ 100, IT < 0.07

Ag-104
69.20
1.15

ECβ+
4.279

Ag-105

41.00
ECβ+
1.346

Ag-106m1

201.84
8.41
EC
3.055

Ag-106
23.96
0.40

ECβ+
2.965
2.965 (ECβ+), 0.195 (β−),

ECβ+ = 99.5, β− < 1

Ag-108
2.37
0.04

β−
1.649
1.649 (β−), 1.918 (ECβ+),

β− = 97.15, ECβ+ = 2.85

Ag-110m1

249.90
β−
3.010
3.010 (β−), 0.188 (IT),

β⁻ = 98.64, IT = 1.,36

Ag-111

178.80
7.45
β−
0.810

Ag-112
187.20
3.12

β−
3.956

Ag-113
322.20
5.37

β−
2.016

Ag-115
20.00
0.33

β−
3.100

Ag-116
2.68

β−
6.160

Cadmium

Cd-102
5.50

ECβ+
2.587

Cd-103
7.30

ECβ+
4.142

Cd-104
57.70
0.96

ECβ+
1.136

Cd-105
55.50

ECβ+
2.739

Cd-107

6.49

ECβ+
1.417

Cd-111
48.54

IT
0.396

Cd-115m1

44.60
β−
1.627

Cd-115

53.46
2.23
β−
1.446

Cd-117m1
201.60
3.36

β−
2.653

Cd-117
149.40
2.49

β−
2.517

Cd-118
50.30

β−
0.520

Cd-119
2.69

β−
3.800

Cd-119m1
2.20

β−
3.947

Indium

In-105
5.07

ECβ+
4.85

In-106
6.20

ECβ+
6.52

In-106m1
5.20

ECβ+
6.55

In-107
32.40

ECβ+
3.43

In-108
58.00

ECβ+
5.15

In-108m1
39.60

ECβ+
5.18

In-109

4.20

ECβ+
2.020

In-110

4.9

ECβ+
3.878

In-110m1
69.10
1.15

ECβ+
3.940

In-111

67.92
2.83
EC
0.245

In-112
14.40
0.24

ECβ+
2.586
2.586 (ECβ+), 0.664 (β−);

ECβ+ = 56, β− = 44

In-113m1

1.66

IT
0.392

In-114m1

49.51
IT
0.190
0.190 (IT), 1.642 (ECβ+),

IT = 96.75, ECβ+ = 3.25

In-115m1

4.49

IT
0.336
0.336 (IT), 0.831 (β−),

IT = 95.0, β⁻ = 5.0

In-116m1
54.15
0.90

β−
3.401

In-117m1
116.50
1.94

β−
1.770
1.770 (β⁻), 0.315 (IT);

β− = 52.9, IT = 47.1

In-117
43.80
0.73

β−
1.455

In-118m1
4.45

β−
4.483

In-119m1
18.00
0.30

β−
2.675
2.675 (β⁻), 0.311 (IT);

β− = 94.4, IT = 5.6

In-119
2.40
0.04

β−
2.364

In-121m1
3.88
0.06

β−
3.674
3.674 (β−), 0.314 (IT),

β = 98.8, IT = 1.2

Tin

Sn-107
2.90

ECβ+
5.01

Sn-108
10.30

ECβ+
2.092

Sn-109
18.00

ECβ+
3.85

Sn-110

4.11

EC
0.638

Sn-111
35.30
0.59

ECβ+
2.445

Sn-113m1

21.40

ECβ+
1.113
0.077 (IT), 1.113 (ECβ+),

IT = 91.1, ECβ+ = 8.9

Sn-113

115.09
ECβ+
1.036

Sn-117m1

13.61
IT
0.135

Sn-119m1

293.00
IT
0.090

Sn-121

27.06
1.13
β−
0.388

Sn-123m1
40.08
0.67

β−
1.429

Sn-123

129.20
β−
1.404

Sn-125

231.36
9.64
β−
2.364

Sn-125m1
9.52

β⁻
2.364

Sn-127

2.10

β−
3.20

Sn-127m1
4.13

β⁻
3.21

Sn-128
59.10
0.99

β−
1.27

Sn-129
2.23

β⁻
4.00

Sn-129m1
6.90

β⁻
4.04
4.035 (β−), 0.035 (IT),

β⁻ ≈ 100, IT ≈ 2 · 10⁻⁴

Sn-130
3.72

β⁻
2.15

Antimony

Sb-113
6.67
0.11

β+
3.905

Sb-114
3.49
0.06

β+
5.880

Sb-155
32.10
0.54

β+
3.030

Sb-116
15.80
0.26

β+
4.707

Sb-116m1
60.30
1.01

β+
5.090

Sb-117
62.80
2.80

β+
1.757

Sb-118
3.60
0.06

β+
3.657

Sb-18m1

5.00

β+
3.907

Sb-119

38.19
1.59
EC
0.594

Sb-120m1

138.24
5.76
EC
2.681

Sb-120
15.89
0.26

ECβ+
2.681

Sb-122

65.28
2.72
β−
1.979
1.979 (β−), 1.620 (ECβ+),

β− = 97.59, ECβ+ = 2.41

Sb-122m2
4.19
0.07

IT
0.164

Sb-124m2
20.20
0.34

IT
0.037

Sb-124

60.20
β−
2.905

Sb-126m1
19.15
0.32

β−
3.688
3.688 (β−), 0.016 (IT),

β− = 86, IT = 14

Sb-126

12.40
β−
3.670

Sb-127

92.40
3.85
β−
1.581

Sb-128

9.01

β⁻
4.380

Sb-128m1
10.40
0.17

β−
4.380
4.380 (β−), 0.0 (IT), β− = 96.4,

IT = 3.6

Sb-129
259.20
4.32

β−
2.380

Sb-129m1
17.70
0.30

β−
4.231
4.231 (β−), 1.851 (IT),

β− = 85, IT = 15

Sb-130
40.00
0.67

β−
4.960

Sb-130m1
6.30
0.11

β⁻
4.960

Sb-131
23.00
0.38

β−
3.190

Sb-132
2.79

β−
5.290

Sb-132m1
4.15
0.07

β⁻
5.290

Sb-133
2.50
0.04

β⁻
4.003

Tellurium

Te-112
2.00

ECβ+
4.35

Te-114
15.20

ECβ+
2.8

Te-115
5.80

ECβ+
4.64

Te-115m1
6.70

ECβ+
4.66
4.66 (ECβ+), 0.02 (IT),

ECβ+ < 100

Te-116

2.49

EC
1.510

Te-117
62.00
1.00

ECβ+
3.535

Te-118
360.00
6

EC
0.278

Te-119
961.80
16.03

ECβ+
2.293

Te-119m1
282.00
4.7

ECβ+
2.554
2.554 (ECβ+), 0.261 (IT),

ECβ+ ≈ 100, IT < 0.008

Te-121m1

154.00
IT
0.294
0.294 (IT), 1.334 (ECB+),

IT = 88.6, ECβ+ = 11.4

Te-121

17.00
EC
1.040

Te-123m1

119.70
IT
0.248

Te-125m1

58.00
IT
0.145

Te-127m1

109.00
IT
0.088
0.088 (IT), 0.786 (β−),

IT = 97.6, β⁻ = 2.4

Te-127

9.35

β−
0.698

Te-129m1

33.60
IT
0.105
0.105 (IT), 1.604 (β−),

IT = 63, β⁻ = 37

Te-129
69.60
1.16

β−
1.498

Te-131m1

30.00
1.25
β−
2.415

Te-131
25.00
0.42

β−
2.233

Te-132

78.20
3.26
β−
0.493

Te-133m1
55.40
0.92

β−
3.254
3.254 (B−), 0.334 (IT),

β⁻ = 82.5, IT = 17.5

Te-133
12.45
0.21

β−
2.920

Te-134
41.80
0.70

β−
1.560

Iodine

I-117
2.22

ECβ+
4.67

I-118
13.70

ECβ+
7.04

I-118m1
8.50

ECβ+
7.14
7.144 (ECβ+), 0.104 (IT),

ECβ+ < 100, IT > 0

I-119
19.10

ECβ+
3.51

I-120m1
53.00
0.88

ECβ+
5.615

I-120
81.00
1.35

ECβ+
5.615

I-121
127.20
2.12

ECβ+
2.270

I-122
3.62
0.06

ECβ+
4.234

I-123

13.20

EC
0.159

I-124

100.32
4.18
β+
2.14

I-125

59.408
EC
0.035

I-126

13.02
ECβ+
2.155
2.155 (ECβ+), 1.258 (β−),

ECβ+ = 56.3, β− = 43.7

I-128
24.99
0.42

β−
2.118
2118 (β−), 1.251 (ECβ+),

β− = 93.1, ECβ+ = 6.9

I-130

12.36

β−
2.949

I-130m1
9.00

IT
0.040
0.040 (IT), 2.989 (β⁻),

IT = 84, β− = 16

I-131

192.96
8.04
β−
0.806

I-132m1
83.60
1.39

IT
0.120
0.120 (IT), 3.697 (β⁻),

IT = 86, β− = 14

I-132

2.30

β−
3.577

I-133

20.80

β−
1.770

I-134
52.60
0.88

β−
4.170

I-134m1
3.60

IT
0.316
0.316 (IT), 4.486 (β⁻),

IT = 97.7, β− = 2.3

I-135

6.61

β−
2.648

Lanthanum

La-127
5.10

ECβ+
4.69

La-127m1
3.70

ECβ+
4.705

La-827
5.00

ECβ+
6.7

La-129
11.60

ECβ+
3.72

La-130
8.70

ECβ+
5.6

La-131
59.00
0.98

ECβ+
2.960

La-132

4.80

ECβ+
4.710

La-132m1
24.30

IT
0.188
0.188 (IT), 4.898 (ECβ+),

IT = 76, ECβ+ = 24

La-133
234.72
3.912

ECβ+
2.23

La-134
6.67
0.11

ECβ+
6.450

La-135

19.50

ECβ+
1.200

La-136
9.87

ECβ+
2.87

La-140

40.27
1.68
β−
3.762

La-141

3.93

β−
2.502

La-142
92.50
1.54

β−
4.505

La-143
14.23
0.24

β−
3.425

Cerium

Ce-129
3.50
0.06

ECβ+
5.05

Ce-130
25.00
0.42

ECβ+
2.2

Ce-131
10.20
0.17

ECβ+
4

Ce-131m1
5.00

ECβ+
4

Ce-132
210.60
3.51

ECβ+
1.29
1.29 (ECβ+), 2.341 (IT)

Ce-133
97.00
1.62

ECβ+
2.9

Ce-133m1
294.00
4.9

ECβ+
2.937

Ce-134

72.00
3.00
EC
0.500

Ce-135

17.60

ECβ+
2.026

Ce-137m1

34.40
1.43
IT
0.254
0.254 (IT), 1.476 (ECβ+),

IT = 99.22, ECβ+ = 0.78

Ce-137
540.00
9.00

EC
1.222

Ce-139

137.66
EC
0.278

Ce-141

32.50
β−
0.581

Ce-143

33.00
1.38
β−
1.462

Ce-144

284.30
β−
0.319

Ce-145
3.01

β⁻
2.54

Ce-146
13.52

β⁻
1.04

Praseodymium

Pr-133
6.50

ECβ+
4.3

Pr-134
17.00

ECβ+
6.2

Pr-134m1
11.00

ECβ+
6.2

Pr-135
24.00

ECβ+
3.72

Pr-136
13.10
0.22

ECβ+
5.126

Pr-137
76.60
1.28

ECβ+
2.702

Pr-138m1

2.10

ECβ+
4.801

Pr-139

4.51

ECβ+
2.129

Pr-140
3.39

ECβ+
3.388

Pr-142m1
14.60
0.24

IT
0.004

Pr-142

19.12

β−
2.162
β− ≈ 100, EC = 0.0164

Pr-143

13.56
β−
0.934

Pr-144m1
7.20
0.12

IT
0.059
IT ≈ 100, β⁻ = 0.07

Pr-144
17.28
0.29

β−
2.997

Pr-145

5.98

β−
1.805

Pr-146
24.15

β⁻
4.2

Pr-147
13.60
0.23

β−
2.69

Pr-148
2.27

β⁻
4.93

Pr-148m1
2.00

β⁻
5.02

Pr-149
2.26

β⁻
3.397

Neodymium

Nd-134
8.50

ECβ+
2.77

Nd-135
12.40

ECβ+
4.8

Nd-135m1
5.50

ECβ+
4.856

Nd-136
50.65
0.84

ECβ+
2.210

Nd-137
38.50

ECβ+
3.69

Nd-138
302.40
5.04

EC
1.1

Nd-139m1
330.00
5.50

ECβ+
3.021
3.021 (ECβ+), 0.231 (IT),

ECβ+ = 88.2, IT = 11.8

Nd-139
29.70
0.50

ECβ+
2.79

Nd-140
202.20
3.37

EC
0.222

Nd-141

2.49

ECβ+
1.823

Nd-147

10.98
β−
0.896

Nd-149

1.73

β−
1.691

Nd-151
12.44
0.21

β−
2.442

Nd-152
11.40

β−
1.11

Promethium

Pm-137
2.40

ECβ+

Pm-138m1
3.24

ECβ+,
6.9

IT

Pm-139
4.15

ECβ+
4.52

Pm-140m1
5.95

ECβ+
6.09

Pm-140m2
5.95

ECβ+

Pm-141
20.90
0.35

ECβ+
3.715

Pm-143

265.00
EC
1.041

Pm-148m1

41.30
β−
2.606
2.606 (β−), 0.138 (IT),

β− = 95.0, IT = 5.0

Pm-148

128.88
5.37
β−
2.468

Pm-149

53.08
2.21
β−
1.071

Pm-150

2.68

β−
3.454

Pm-151

28.40
1.18
β−
1.187

Pm-152
4.12

β⁻
3.5

Pm-152m1
7.52

β⁻
3.56

Pm-152m2
13.80

β⁻

β⁻ < 100, IT > 0

Pm-153
5.25

β⁻
1.9

Pm-154m1
2.68

β⁻
4.05

Samarium

Sm-138
3.10
0.05

ECβ+
3.900

Sm-139
2.57
0.04

ECβ+
5.460

Sm-140
14.80
0.25

ECβ+
3.020

Sm-141m1
22.60
0.38

ECβ+
4.719
4.719 (ECβ+), 0.176 (IT);

ECβ+ = 99.69, IT = 0.31

Sm-141
10.20
0.17

ECβ+
4.543

Sm-142
72.49
1.21

ECβ+
2.090

Sm-143
8.83

ECβ+
3.443

Sm-145

340.00
EC
0.617

Sm-153

46.80
1.95
β−
0.810

Sm-155
22.30
0.37

β−
1.627

Sm-156

9.40

β−
0.722

Sm-158
5.30
0.09

β−
1.999

Europium

Eu-143
2.63

ECβ+
5.275

Eu-145

142.56
5.94
ECβ+
2.660

Eu-146

110.64
4.61
ECβ+
3.878

Eu-147

24.10
ECβ+
1.722

Eu-148

54.50
ECβ+
3.107

Eu-149

93.10
EC
0.692

Eu-150

12.62

β−
1.013
ECB+ = 11, β− = 89,

IT < 5 · 10-8

Eu-152m1

9.32

β−
1.865
ECβ+ = 28,β− = 72,

1.920 (ECβ+), 1.865 (β−)

Eu-152m2
96.00
1.6

IT
0.148

Eu-154m1
46.30
0.77

IT
0.145

Eu-156

15.19
β−
2.451

Eu-157

15.15

β−
1.363

Eu-158
45.90
0.77

β−
3.490

Eu-159
18.10

β⁻
2.514

Gadolinium

Gd-144
4.50

ECβ+
3.74

Gd-145
22.90
0.38

ECβ+
5.050

Gd-146

48.30
EC
1.030

Gd-147

38.10
1.59
ECβ+
2.187

Gd-149

225.60
9.40
ECβ+
1.314

Gd-151

120.00
EC
0.464

Gd-153

242.00
EC
0.485

Gd-159

18.49

β−
0.971

Gd-161
3.66

β⁻
1.956

Gd-162
8.40

β⁻
1.39

Terbium

Tb-147

1.65

ECβ+
4.609

Tb-148

1.00

ECβ+
5.690

Tb-148m1
2.20

ECβ+
5.78

Tb-149

4.15

β+
2.62
3.636 (ECβ+), 4.113 (α);

ECβ+ = 83.3, α = 16.7

Tb-149m1
4.16

EC+
3.672
3.672 (ECβ+), 4.077 (α),

ECβ+ = 99.978, α = 0.022

Tb-150

3.27

EC+
4.656

Tb-150m1
5.80

EC+
5.13

Tb-151

17.60

β+
1.54
2.565 (ECβ+), 3.497 (α);

ECβ+ = 100, α = 9.5 · 10-3

Tb-152m1
4.20

IT
0.052
0.502 (IT), 4.492 (ECβ+),

IT = 78.8, ECβ+ = 21.2

Tb-152

17.50

ECβ+
3.990
3,.990 (ECβ+), 3.090 (α);

ECβ+ = 100, a < 7 · 10-7

Tb-153

56.16
2.34
ECβ+
1.570

Tb-154

21.40

ECβ+
3.560
3.56 (ECβ+), 0.25 (β−),

EC+ = 100, β− < 0.1

Tb-154m1

9.4

ECβ+
3.560
3.56 (ECβ+), 0.0(IT), 0.25

(β−), ECβ+ = 78.2, IT = 21.8,

β− < 0.1

Tb-154m2

22.7

ECβ+
3.560
3.56 (ECβ+), 0.0 (IT),

ECβ+ = 98.2, IT = 1.8

Tb-155

127.68
5.32
EC
0.821

Tb-156m1

24.40

IT
0.050

Tb-156m2

5.00

IT
0.088
0.088 (IT), 2.532 (ECB+)

Tb-156

128.40
5.35
ECβ+
2.444
2.444 (ECβ+), 0.434 (β−);

ECβ+ ≈ 100, β− = ?

Tb-160

72.30
β−
1.835

Tb-161

165.84
6.91
β−
0.593

Tb-162
7.60
0.13

β−
2.510

Tb-163
19.50
0.33

β−
1.785

Tb-164
3.00

β⁻
3.89

Tb-165
2.11

β⁻
3

Dysprosium

Dy-148
3.10
186

ECβ+
2.678

Dy-149
4.20
252

ECβ+
3.812

Dy-150
7.17
430.2

ECβ+
1.794
4.351(α), 1.794 (ECβ+),

α = 36, ECβ+ = 64

Dy-151
17.90

ECβ+
2.870
2.87 (ECβ+), 4.180 (α),

ECβ+ = 94.4, α = 5.6

Dy-152

2.38

ECβ+
0.600
0.60 (ECβ+), 3.727 (α),

EC(?) = 99.900, α = 0.100

Dy-153

6.4

ECβ+
2.170
2.17 (ECβ+), 3.559 (α),

ECβ+= 100, α = 0.0094

Dy-155

9.90

ECβ+
2.095

Dy-157

8.14

ECβ+
1.341

Dy-159

144.40
EC
0.366

Dy-165

2.33

β−
1.290

Dy-166

81.60
3.40
β−
0.486

Dy-167
6.20

β⁻
2.35

Dy-168
8.70

β⁻
1.6

Holmium

Ho-153
2.01

ECβ+
4.129
4.129 (ECβ+), 4.015 (α),

ECβ+ = 99.949, α = 0.051

Ho-153m1
9.30

ECβ+
4.179
4.179 (ECβ+), 4.119 (α),

ECβ+ = 99.82, α = 0.18

Ho-154
11.76

ECβ+
5.751
5.751 (ECβ+), 4.042 (α),

ECβ+ = 99.981, α = 0.019

Ho-154m1
3.10

ECβ+
6.071
6.071 (ECβ+), 4.362 (α),

0.320 (IT), ECβ+ = 100,

α < 0.001, IT ≈ 0

Ho-155
48.00
0.80

ECβ+
3.102

Ho-156
56.00
0.93

ECβ+
5.060

Ho-157
12.60
0.21

ECβ+
2.540

Ho-158
11.30

ECβ+
4.23

Ho-158m1
28.00

IT
0.067
4.297 (ECβ+), 0.067 (IT),

ECβ+ < 19, IT > 81

Ho-158m2
21.30

ECβ+
4.410
4.41 (ECβ+), 0.18 (IT),

ECβ+ > 93, IT < 7

Ho-159
33.00
0.55

ECβ+
1.838

Ho-160
25.60

ECβ+
3.29

Ho-160m1
301.20
5.02

IT
0.060
0.06 (IT), 3.35 (ECβ+),

IT = 65, ECβ+ = 35

Ho-161
150.00
2.50

EC
0.895

Ho-162m1
67.00
1.12

IT
0.106
0.106 (IT), 2.246 (ECβ+),

IT = 62, ECβ+ = 38

Ho-162
15.00
0.25

ECβ+
2.140

Ho-164m1
37.50
0.63

IT
0.140

Ho-164
29.00
0.48

EC
0.987
0.987 (EC), 0.962 (β−);

EC = 60, β− = 40

Ho-166

26.80
1.12
β−
1.855

Ho-167

3.10

β−
1.007

Ho-168
2.99

β⁻
2.91

Ho-169
4.70

β⁻
2.124

Ho-170
2.76

β⁻
3.87

Erbium

Er-154
3.73

ECβ+
2.032
2.032 (ECβ+), 4.280 (α),

ECβ+ = 99.53, α = 0.47

Er-155
5.30

ECβ+
3.84
3.84(ECβ+), 4.12(α),

ECβ+ = 99.978, α = 0.022

Er-156
19.50

ECβ+
1.37

Er-157
18.65

ECβ+
3.5
3.50 (ECβ+), 3.30(α),

ECβ+ ≈ 100, α < 0.02

Er-158
137.40
2.29

EC
0.9

Er-159
36.00

ECβ+
2.769

Er-160

28.58

EC
0.33

Er-161
192.60
3.21

ECβ+

Er-163
75.00
1.25

ECβ+
1.21

Er-165
621.60
10.36

EC
0.376

Er-169

223.20
9.30
β−
0.340

Er-171
451.20
7.52

β−
1.490

Er-172

49.30
2.05
β−
0.891

Er-174
3.30

β⁻
1.8

Thulium

Tm-157
3.63

ECβ+
4.48

Tm-158
3.98

ECβ+
6.6

Tm-159
9.13

ECβ+
3.85

Tm-160
9.40

ECβ+
5.6

Tm-161
33.00

ECβ+
3.16

Tm-162
21.70
0.36

ECβ+
4.810

Tm-163
108.60
1.81

ECβ+
2.439

Tm-164
2.00

ECβ+
3.962

Tm-164m1
5.10

ECβ+
3.962

Tm-165

30.06

ECβ+
1.592

Tm-166
462.00
7.70

ECβ+
3.040

Tm-167

221.76
9.24
EC
0.748

Tm-168

93.10
ECβ+
1.679
1.679 (ECβ+), 0.257(β−),

ECβ+ = 99.990, β− = 0.010

Tm-170

128.60
β−
0.968
0.314 (ECβ+), 0.968 (β−),

EC, β−(99%)

Tm-172

63.60
2.65
β−
1.880

Tm-173

8.24

β−
1.298

Tm-174
5.40

β⁻
3.08

Tm-175
15.20
0.25

β−
2.39

Tm-176
1.90

β⁻
3.88

Ytterbium

Yb-160
4.80

ECβ+
2.3

Yb-161
4.20

ECβ+
4.15

Yb-162
18.90
0.32

EC
1.660

Yb-163
11.05

ECβ+
3.37

Yb-164
75.80

EC
1

Yb-165
9.90

ECβ+
2.762

Yb-166

56.70
2.36
EC
0.304

Yb-167
17.50
0.29

EC+
1.954

Yb-169

32.01
EC
0.909

Yb-175

100.56
4.19
β−
0.47

Yb-177

1.90

β−
1.399

Yb-178
74.00
1.23

β−
0.645

Yb-179
8.00

β⁻
2.4

Yb-180
2.40

β⁻

Lutetium

Lu-162m2
1.90

ECβ+

Lu-164
3.14

ECβ+
6.25

Lu-165
10.74

ECβ+
3.92

Lu-166
2.65

ECβ+
5.48

Lu-166m2
2.12

ECβ+
5.523
5.523 (EC+), 0.043

(IT), ECβ+ > 80, IT < 20

Lu-167
51.50
0.86

ECβ+
3.130

Lu-168
5.50

ECβ+
4.48

Lu-168m1
6.70

ECβ+
4.700
4.70 (ECβ+), 0.220 (IT),

ECβ+ > 95, IT < 5

Lu-169

34.06
1.42
ECβ+
2.293

Lu-170

48.00
2.00
ECβ+
3.459

Lu-171

197.28
8.22
ECβ+
1.479

Lu-172

160.80
6.70
ECβ+
2.519

Lu-174m1

142.00
IT
0.171
0.171 (IT), 1.545 (EC),

IT = 99.38, EC = 0.62

Lu-176m1

3.68

β−
1.316
1.316(β−), 0.229 (EC),

β− = 99.905, EC = 0.095

Lu-177m1

160.90
β−
1.468
1.468 (β−), 0.970 (IT),

β− = 78.3, IT = 21.7

Lu-177

6.71
β−
0.490

Lu-178m1
22.70
0.38

β−
2.219

Lu-178
28.40
0.47

β−
2.099

Lu-179

4.59

β−
1.405

Lu-180
5.70

β⁻
3.1

Lu-181
3.50

β⁻
2.5

Lu-182
2.00

β⁻

Hafnium

Hf-166
6.77

ECβ+
2.3

Hf-167
2.05

ECβ+
4

Hf-168
25.95

ECβ+
1.8

Hf-169
3.24

ECβ+
3.27

Hf-170

16.01

EC
1.1

Hf-171

12.1

ECβ+
2.4

Hf-173

23.60
0.98
ECβ+
1.610

Hf-175

70.00
EC
0.686

Hf-177m1
51.40
0.86

IT
2.740

Hf-179m2

25.10
IT
1.106

Hf-180m1

5.50

IT
1.141
1.141 (IT), 1.287(β−),

IT = 99,.7, β− = 0.3

Hf-181

42.40
β−
1.027

Hf-182m1
61.50
1.03

β−
1.546
1.546 (β−), 1.173 (IT),

β− = 58, IT = 42

Hf-183
64.00
1.07

β−
2.010

Hf-184

4.12

β−
1.340

Hf-185
3.50

β⁻

Tantalum

Ta-168
2.00

ECβ+
6.7

Ta-169
4.90

ECβ+
4.44

Ta-170
6.76

ECβ+
6

Ta-171
23.30

ECβ+
3.7

Ta-172
36.80
0.61

ECβ+
4.920

Ta-173

3.65

ECβ+
2.790

Ta-174

1.20

ECβ+
3.850

Ta-175

10.50

ECβ+
2.000

Ta-176

8.08

ECβ+
3.110

Ta-177

56.60
2.36
EC
1.166

Ta-178m1

2.36

EC
1.910

Ta-178
9.31
0.16

EC
1.910

Ta-180

8.15

EC
0.854
0.854 (EC), 0.708 (β−),

EC = 86, β− = 14

Ta-182m1
15.84
0.26

IT
0.52

Ta-182

115.00
β−
1814.000

Ta-183

122.40
5.10
β−
1.070

Ta-184

8.70

β−
2.870

Ta-185
49.00
0.82

β−
1.992

Ta-186
10.50
0.18

β−
3.000

Tungsten

W-170
2.42

ECβ+
3

W-171
2.38

ECβ+
4.6

W-172
6.60

ECβ+
2.5

W-173
7.60

ECβ+
4

W-174
31.00

ECβ+
1.9

W-175
35.20

ECβ+
2.91

W-176

2.50

EC
0.790

W-177
135.00
2.25

ECβ+
2.000

W-178

21.70
EC
0.091

W-179m1
6.40

IT
0.222
0.222 (IT), 1.282 (ECβ+),

IT = 99.72, ECβ+ = 0.28

W-181

121.20
EC
0.188

W-185

75.10
β−
0.433

W-187

23.72
0.99
β−
1.311

W-188

69.40
β−
0.349

W-189
11.50

β⁻
2.5

W-190
30.00

β⁻
1.27

Rhenium

Re-173
1.98

ECβ+
4.8

Re-174
2.40

ECβ+
6.5

Re-175
5.89

ECβ+
4.3

Re-176
5.30

ECβ+
5.6

Re-177
14.00
0.23

ECβ+
3.400

Re-178
13.20
0.22

ECβ+
13.200

Re-179
19.50

ECβ+
2.71

Re-180
2.43
0.04

ECβ+
3.800

Re-181

20.00

ECβ+
1.739

Re-182

64.00

EC
2.800

Re-182m1

12.70

ECβ+
2.800

Re-183

70.00
EC
0.556

Re-184m1

169.00
IT
0.188
0.188 (IT), 1.671 (EC),

IT = 75.4, EC = 24.6

Re-184

38.00
ECβ+
1.483

Re-186

90.48
3.72
β−
1.07
0.582 (EC), 1.069 (β−);

EC = 7.47, β− = 92.53

Re-188m1
18.60
0.31

IT
0.172

Re-188

16.98

β−
2.120

Re-189

24.30
1.01
β−
1.009

Re-190
3.10

β−
3.15

Re-190m1
192.00
3.2

β⁻
3.269
3.269 (β−), 0.119 (IT),

β⁻ = 54.4, IT = 45.6

Re-191
9.80

β⁻
2.045

Osmium

Os-176
3.60

ECβ+
3.2

Os-177
2.80

ECβ+
4.5

Os-178
5.00

ECβ+
2.3

Os-179
6.50

ECβ+
3.68

Os-180
22.00
0.37

ECβ+
1.470

Os-181
105.00
1.75

ECβ+
2.930

Os-181m1
2.70

ECβ+
2.979

Os-182

22.00

EC
0.91

Os-183
13.00

ECβ+
2.13

Os-183m1
9.90

ECβ+
2.301
2.301 (ECβ+), 0.171 (IT),

ECβ+ = 85, IT = 15

Os-185

94.00
EC
1.013

Os-189m1

6.00

IT
0.031

Os-190m1
9.90
0.17

IT
1.705

Os-191m1

13.03

IT
0.074

Os-191

15.40
β−
0.314

Os-193

30.00
1.25
β−
1.140

Os-195
6.50

β⁻
2

Os-196
34.90

β⁻
1.16

Iridium

Ir-181
4.90

ECβ+
4.07

Ir-182
15.00
0.25

ECβ+
5.61

Ir-183
58.00

ECβ+
3.45

Ir-184

3.02

ECβ+
4.600

Ir-185

14.00

ECβ+
2.370

Ir-186

15.80

ECβ+
3.831

Ir-186m1

1.90

ECβ+
3.831
3.831 (ECβ+), 0 (IT),

ECβ+ ≈ 75, IT ≈ 25

Ir-187

10.50

EC
1.502

Ir-188

41.50
1.73
ECβ+
2.809

Ir-189

13.30
EC
0.532

Ir-190m2

3.25

ECβ+
2.149
2.149 (ECβ+), 0.140 (IT),

ECβ+ = 94.4, IT = 5.6

Ir-190m1

1.20

IT
0.026

Ir-190

12.10
ECβ+
2.000

Ir-192

73.83
β−
1.460
1.46 (β−), 1.046 (EC),

β− = 95.24, EC = 4.76

Ir-193m1

10.53
IT
0.08

Ir-194m1

171.00
β−
2.437

Ir-194

19.15

β−
2.247

Ir-195m1

3.80

β−
1.220
1.22 (β−), 0.10 (IT),

β− = 95, IT = 5

Ir-195

2.50

β−
1.120

Ir-196m1
84.00
1.4

β−
3.620

Ir-197
5.80

β−
2.155

Ir-197m1
8.90

β−
2.270
2.27 (β−), 0.115 (IT),

β⁻ = 99.75, IT = 0.25

Platinum

Pt-182
3.00

ECβ+
2.850
2.85 (ECβ+), 4.943 (α),

ECβ+ = 99.969, α = 0.031

Pt-183
6.50

ECβ+
4.600

Pt-184
17.30

ECβ+
2.300

Pt-185
70.90
1.1817

ECβ+
3.800

Pt-185m1
33.00

EC+
3.903
3.903 (ECβ+), 0.103 (IT),

4.643(α), ECβ+ = 99, IT < 2

Pt-186

2.00

EC+
1.380

Pt-188

10.20
EC
0.507

Pt-187

2.35

ECβ+
3.11

Pt-189

10.87

ECβ+
1.971

Pt-191

67.20
2.80
EC
1.019

Pt-193m1

103.92
4.33
IT
0.150

Pt-195m1

96.48
4.02
IT
0.259

Pt-197m1
95.41
1.59

IT
0.399
0.399 (IT) 1.119 (β−),

IT = 96.7, β− = 3.3

Pt-197

18.30

β−
0.719

Pt-199
30.80
0.51

β−
1.702

Pt-200

12.50

β−
0.660

Pt-201
2.50

β⁻
2.66

Pt-202

44

β⁻

Gold

Au-185
4.25

ECβ+
4.71
4.71 (ECβ+), 5.18 (α),

ECβ+ = 99.74, α = 0.26

Au-185m1
6.80

ECβ+
4.71

Au-186
10.70

ECβ+
6.04

Au-187
8.40

ECβ+
3.6
3.6 (ECβ+), 4,79 (α),

ECβ+ = 99.997, α = 0.003

Au-188
8.84

ECβ+
5.3

Au-189
28.70

ECβ+
2.85
EC+ = 100, α < 3 ·1 0-5

Au-189m1
4.59

ECβ+
3.097
ECβ+ = 100, IT > 0

Au-190
42.80

ECβ+
4.442
EC+= 100, α < 1 · 10-6

Au-191
3.18

ECβ+
1.83

Au-192
4.94

ECβ+
3.516

Au-193

17.65

EC
1.069

Au-194

398.02
16.58
ECβ+
2.492

Au-195

183.00
EC
0.227

Au-196

148.39
6.18
ECβ+
1.500
1.506 (ECβ+), 0.686 (β−),

ECβ+ = 92.80, β− = 7.20

Au-196m2

9.60

IT
0.596

Au-198m1

55.20
2.30
IT
0.812

Au-198

64.70
2.70
β−
1.372

Au-199

75.34
3.14
β−
0.453

Au-200m1

18.70

β−
3.202
3.202 (β−), 0.962 (IT),

β− = 82, IT = 18

Au-200
48.40
0.81

β−
2.240

Au-201
26.40
0.44

β−
1.275

Thallium

TI-189
2.30

EC+
5.18

TI-190
2.60

EC+
7

TI-190m1
3.70

EC+
7

TI-191

EC+
4.49

TI-191m1
5.22

EC+
4.789

TI-192
9.60

EC+
6.12

TI-192m1
10.80

EC+
6.12

TI-193
21.60

EC+
3.64

TI-193m1
2.11

IT
0.365
0.365 (IT), 4.005 (ECβ+),

IT = 75, ECβ+ = 25

TI-194m1
32.80
0.55

EC+
5.280

TI-194
33.00
0.55

EC
5.280

TI-195

1.16

ECβ+
2.810

TI-196

1.84

ECβ+
4.38

TI-196m1

1.41

ECβ+
4.774
4.774 (ECβ+) 0.394 (IT),

ECβ+ = 95.5, IT = 4.5

TI-197

2.84

ECβ+
2.180

TI-198m1

1.87

ECβ+
4.004
4.004 (ECβ+), 0.544 (IT),

ECβ+ = 54, IT = 46

TI-198

5.30

ECβ+
3.460

TI-199

7.42

ECβ+
1.440

TI-200

26.10
1.09
ECβ+
2.456

TI-201

3.04
EC
0.483

TI-202

12.23
ECβ+
1.365

TI-206
4.20
0.07

β−
1.533

TI-206m1
3.74

IT
2.643

TI-207
4.77
0.08

β−
1.423

TI-208
3.07
0.05

β−
5.001

TI-209
2.20
0.04

β−
3.980

Lead

Pb-191m1
2.10

ECβ+
6.038

Pb-192
3.50

ECβ+
3.400
3.4 (ECβ+), 5.221 (α),

ECβ+ = 99.9941,

α = 0.0059

Pb-193
2.00

ECβ+

Pb-193m1
5.80

ECβ+
5.200

Pb-194
12.00

ECβ+
2.700

Pb-195
15.00

ECβ+
4.500

Pb-195m1
15.80
0.26

ECβ+
4.500

Pb-196
37.00

ECβ+
2.050
2.05 (ECβ+), 4.2 (α),

ECβ+ ≈ 100, α < 3 · 10-5

Pb-197
8.00

ECβ+
3.58

Pb-197m1
43.00

ECβ+
3.889
3.889 (ECβ+), 0.319 (IT),

ECβ+ = 81, IT = 19

Pb-198

2.40

EC+
1.410

Pb-199
90.00
1.50

EC+
2.880

Pb-199m1
12.20

IT
0.425
0.425 (IT), 3.305 (ECβ+),

IT = 93, ECβ+ = 7

Pb-200

21.50

EC
0.811

Pb-201

9.33

ECβ+
1.900

Pb-202m1

3.53

IT
2.710

Pb-203

51.87
2.16
EC
0.975

Pb-204m1
67.20
1.12

IT
2.186

Pb-209

3.25

β−
0.644

Pb-211
36.10
0.60

β−
1.373

Pb-212

10.64

β−
0.574

Pb-213
10.20
0.17

β−
2.070

Pb-214
26.80
0.45

β−
1.024

Bismuth

Bi-197
9.33

ECβ+
5.200
5.2 (ECβ+), 5.39 (α),

ECβ+ ≈ 100, α = 1 · 10-4

Bi-197m1
5.04

α
5.890
5.89 (α), 5.7 (ECβ+), 0.50

(IT), α = 55, ECβ+ = 45,

IT < 0.3

Bi-198
10.30

ECβ+
6.56

Bi-198m1
11.60

ECβ+
6.56

Bi-199
27.00

ECβ+
4.34

Bi-199m1
24.70

ECβ+
5.020
5.02 (ECβ+), 5.64 (α),

0.68 (IT), ECβ+ = 99,

α ≈ 0.01, IT < 2

Bi-200
36.40

ECβ+
5.89

Bi-200m1
31.00

ECβ+
5.89
ECβ+ > 90, IT < 10

Bi-201
108.00
1.80

EC
3.84

Bi-201m1
59.10
0.99

EC
4.686
4.686 (EC), 5.346 (IT),

5.346 (α), EC > 93, IT < 6.8,

α ≈ 0.3

Bi-202

1.67

ECβ+
5.150
5.15 (ECβ+), 4.29 (α),

ECβ+ = 100, α < 1·10-5

Bi-203

11.76

ECβ+
3.253
3.253 (ECβ+), 4.15 (α),

ECβ+ = 100, α ≈ 1 · 10-5

Bi-204

11.22

ECβ+
4.438

Bi-205

15.31
ECβ+
2.708

Bi-206

149.83
6.24
ECβ+
3.758

Bi-210

120.29
5.01
β−
1.163

Bi-211
2.14
0.04

α
6.751
6.751 (α), 0.579 (β−),

α = 99.724, β− = 0.276

Bi-212
60.55
1.01

β−
2.254
2.254 (β⁻), 6.207 (α),

11.208 (β⁻ + α); β− = 64.06,

α = 35.94

Bi-212m1
25.00

α
6.457
6.457 (α), 2.504 (β−),

α = 67, β⁻ = 33, β⁻α = 30

Bi-212m2
7.00

β−
4.164

Bi-213
45.6
0.76

α
5.98
1.464 (β⁻), 5.982 (α);

β− = 97.91, α = 2.09

Bi-214
19.90
0.33

β−
3.272
3.272 (β⁻), 5.617 (α);

β− = 99.979, α = 0.021

Bi-215
7.60

β−
2.25

Bi-216
3.60

β−
4

Polonium

Po-199
5.48

ECβ+
5.600
5.6 (ECβ+), 6.074 (α),

ECβ+ = 88, α = 12

Po-199m1
4.13

ECβ+
5.910
5.91 (ECβ+), 6.384 (α),

0.310 (IT), ECβ+ = 59,

α = 39, IT = 2.1

Po-200
11.50

ECβ+
3.350
3.35 (ECβ+), 5.982 (α),

ECβ+ = 88.9, α = 11.1

Po-201
15.30

ECβ+
4.880
4.88 (ECβ+), 5.799 (α),

ECβ+ = 98.4, α = 1.6

Po-201m1
8.90

IT
0.424
0.424 (IT), 5.304 (ECβ+),

6.223 (α), IT = 56,

EC = 41, α ≈ 2.9

Po-202
44.70
0.75

ECβ+
2.820
2.82 (ECβ+), 5.701 (α),

ECβ+ = 98.08, α = 1.92

Po-203
36.70
0.61

ECβ+
4.230
4.23 (ECβ+), 5.496 (α),

ECβ+ = 99.89, α = 0.11

Po-204

3.53

ECβ+
2.340
2.34 (ECβ+), 5.485 (α),

ECβ+ = 99.34, α = 0.,66

Po-205

1.66

ECβ+
3.530
3.53 (ECβ+), 5.324 (α),

ECβ+ = 99.96, α = 0.04

Po-206

8.8

ECβ+
1.846
1.846 (ECβ+), 5.326(α),

ECβ+ = 94.55, α = 5.45

Po-207

5.8

ECβ+
2.909
2.909 (ECβ+), 5.216 (α),

ECβ+ = 99.979, α = 0.021

Po-210

138.38
α
5.307

Po-218
3.05
0.05

α
6.115
6.115 (α), 0.265 (β−),

α = 99.980, β⁻ = 0.020

Astatine

At-203
7.40

ECβ+
5.060
5.06 (ECβ+), 6.21 (α),

ECβ+ = 69, α = 31

At-204
9.20

EC+
6.480
6.48 (ECβ+), 6.07 (α),

ECβ+ = 96.2, α = 3.8

At-205
26.20
0.44

EC+
4.540
4.54 (ECβ+), 6.02 (α),

ECβ+ = 90, α = 10

At-206
30.00
0.50

EC+
5.720
5.72 (ECβ+), 5.888 (α),

ECβ+ = 99.11, α = 0.89

At-207

1.80

EC+
3.910
3.91 (ECβ+), 5.873 (α),

ECβ+ = 91.4, α = 8.6

At-208

1.63

EC+
4.973
4.973 (ECβ+), 5.751 (α),

ECβ+ = 99.45, α = 0.55

At-209

5.41

EC+
3.486
3.486 (ECβ+), 5.757 (α),

ECβ+ = 95.9, α = 4.1

At-210

8.1

EC+
3.981
3.981 (ECβ+), 5.631 (α),

ECβ+ = 99.825, α = 0.175

At-211

7.21

α+
5.98
0.786 (ECβ+), 5.982 (α),

EC = 58.2, α = 41.8

At-220
3.71

β−

α = 8, β− = 92, 3.65

(ECβ+), 6.05 (α)

At-221
2.30

β−

Radon

Rn-205
2.80

ECβ+
5.240
5.24 (ECβ+), 6.39 (α),

ECβ+ = 77, α = 23

Rn-206
5.67

α
6.384
6.384 (α), 3.,31 (ECβ+),

α = 63, ECβ+ = 37

Rn-207
9.25

ECβ+
4.610
4.61 (ECβ+), 6.251 (α),

ECβ+ = 79, α = 21

Rn-208
24.35
0.41

α
6.260
6.26 (α), 2.85 (ECβ+),

α = 62, ECβ+ = 38

Rn-209
28.50
0.48

ECβ+
3.930
3.93 (ECβ+), 6.155 (α),

ECβ+ = 83, α = 17

Rn-210

2.40

α
6.159
6.159(α), 2.374 (ECβ+),

α = 96, ECβ+ = 4

Rn-211

14.60

EC
2.892
2.892 (ECβ+), 5.965(α),

EC = 72.6, α = 27.4

Rn-212
23.90
0.40

α
6.385

Rn-221
25.00

β−
1.220
1.22 (β−), 6.146 (α),

β⁻ = 78, α = 22

Rn-222

3.82
α
5.590

Rn-223
23.20

β−

β⁻ ≈ 100, α = 0.0004

Rn-224
107.00

β⁻
0.8

Rn-225
4.50

β⁻

Rn-226
7.40

β⁻
1.4

Francium

Fr-210
3.18

α
6.700
6.7 (α), 6.262 (ECβ+),

α = 60, ECβ+ = 40

Fr-211
3.10

α
6.660
6.66 (α), 4.605 (ECβ+),

α > 80, EC < 20

Fr-212
20.00
0.33

EC+
5.117
5.117 (ECβ+), 6.529 (α),

ECβ+ = 57, α = 43

Fr-221
4.80
0.08

α
6.458
α ≈ 100, β⁻ = ?,

¹⁴C = 8.8 · 10

Fr-222
14.40
0.24

β−
2.033

Fr-223
21.80
0.36

β−
1.149

Fr-224
3.33

β⁻
2.83

Fr-225
4.00

β⁻
1.866

Fr-227
2.47

β⁻
2.49

Radium

Ra-213
2.74

α
6.859
6.859 (α), 3.88 (ECβ+),

α = 80, ECβ+ = 20

Ra-223

11.43
α
5.979

Ra-224

87.84
3.66
α
5.789

Ra-225

14.80
β−
0.357

Ra-227
42.20
0.70

β−
1.325

Ra-229
4.00

β⁻
1.76

Ra-230
93.00
1.55

β−
0.990

Actinium

Ac-223
2.10
0.04

α
6.783

Ac-224

2.90

α
1.403
1.403 (EC), 6.327 (α),

0.232 (β−), EC = 90.9,

α = 9.1, β− < 1.6

Ac-225

10.00
α
5.935

Ac-226

29.00
1.21
β−
1.117
1.117 (β⁻), 0.64 (EC),

5.563 (α), β− ≈ 83,

EC = 17, α = 6 · 10-3

Ac-228

6.13

β−
2.127

Ac-229
62.70

β⁻
1.1

Ac-231
7.50

β⁻
2.1

Thorium

Th-225
8.72

α
6.922
6.922 (α), 0.675 (EC),

α = 90, EC ≈ 10

Th-226
30.90
0.52

α
6.451

Th-227

18.72
α
6.051

Th-231

25.52
1.06
β−
0.389

Th-233
22.30

β⁻
1.245

Th-234

24.10
β−
0.273

Th-235
7.10

β⁻
1.93

Th-236
37.00
0.62

β⁻

Th-237
5.00

β⁻

Protactinium

Pa-227
38.30
0.64

α
6.580
6.580 (α), 1.019 (EC),

α = 85, EC = 15

Pa-228

22.00

EC+
2.148
2.148 (ECβ+), 6.265 (α),

ECβ+ = 98.0, α = 2.0

Pa-229

36.00
1.50
EC
0.316

Pa-230

17.40
ECβ+
1.310
1.310 (ECβ+), 0.563 (β−),

5.439 (α), ECβ+ = 91.6,

β− = 8.4, α = 0.0032

Pa-232

31.44
1.31
β−
1337.000

Pa-233

27.00
β−
0.571

Pa-234

6.70

β−
2.197

Pa-235
24.50

β⁻
1.41

Pa-236
9.10

β⁻
2.9

Pa-237
8.70

β⁻
2.25

Pa-238
2.30

β⁻
3.46

Uranium

U-228
9.10

α
6.801
6.804 (α), 0.307 (EC),

α > 95, EC < 5

U-229
58.00
0.97

ECβ+
1.309
1.309 (ECβ+), 6.475(α),

ECβ+ ≈ 80, α ≈ 20

U-230

20.80
α
5.993

U-231

100.80
4.20
EC
0.360

U-235m1
25.00

IT

U-237

162.00
6.75
β−
0.519

U-239
23.54
0.39

β−
1.265

U-240

14.10

β−
0.338

U-242
16.80

β⁻

Neptunium

Np-229
4.00

α
2.560
7.01 (α), 2.56 (EC),

α > 50, EC < 50

Np-230
4.60

ECβ+
3.610
3.,61(ECβ+), 6.78 (α),

ECβ+ < 97, α > 3

Np-231
48.80

ECβ+
1.840
1.84 (EC), 6.37 (α),

EC = 98, α = 2

Np-232
14.70
0.25

ECβ+
2.700

Np-233
36.20
0.60

EC
1.230

Np-234

105.60
4.40
ECβ+
1.810

Np-236m1

22.50

EC
1.000
1.00 (EC), 0.55 (β−),

EC = 52, β− = 48

Np-238

50.81
2.12
β−
1.292

Np-239

56.52
2.36
β−
0.722

Np-240m1
7.40
0.12

β−
2.200

Np-240
65.00
1.08

β−
2.200

Np-241
13.90

β⁻
1.31

Np-242
5.50

β⁻
2.7

Np242m1
2.20

β⁻
2.7

Np-244
2.29

β⁻

Plutonium

Pu-231
8.60

EC+, α

Pu-232
34.10

ECβ+
1.06
1.06 (ECβ+), 6.716 (α),

EC = 77, α = 23

Pu-233
20.90
0.35

ECβ+
1.900

Pu-234

8.80

EC
0.388
0.388 (EC+), 6.31(α),

EC ≈ 94, α ≈ 6

Pu-235
25.30
0.42

ECβ+
1.170

Pu-237

45.30
EC
0.220

Pu-243

4.96

β−
0.528

Pu-245

10.50

β−
1.205

Pu-246

10.85
β−
0.401

Pu-247

2.27
β⁻

Americium

Am-234
2.32

EC = 100, α = 0.039,

ECSF = 0.0066

Am-235
15.00

Am-237
73.00
1.22

EC
1.730

Am-238
98.00
1.63

EC
2.260

Am-239

11.90

EC
0.803

Am-240

50.80
2.12
EC
1.379

Am-242

16.02

β−
0.665
0.665 (β−), 0.751 (EC),

β− = 82.7, EC = 17.3

Am-244m1
26.00
0.43

β−
1.516

Am-244

10.10

β−
1.428

Am-245

2.05

β−
0.894

Am-246m1
25.00
0.42

β−
2.376

Am-246
39.00
0.65

β−
2.376

Am-247
23.00

β⁻
1.7

Am-248

β⁻
3.1

Curium

Cm-236
10.00

ECβ+
1.710

Cm-237
20.00

Cm-238

2.40

EC
0.970
0.97 (EC), 6.62 (α),

EC = 96.16, α = 3.84

Cm-239

2.90

EC
1.700

Cm-240

27.00
a
6.397

Cm-241

32.80
EC
0.767

Cm-242

162.80
a
6.216

Cm-249
64.15
1.07

β−

Cm-251
16.80

β⁻
1.42

Cm-252

2
β⁻

Berkelium

Bk-240
4.80

ECβ+
3.94

Bk-242
7.00
0.12

ECβ+
3.000

Bk-243

4.50

EC
1.508

Bk-244

4.35

EC
2.260

Bk-245

118.56
4.94
EC
0.810

Bk-246

43.92
1.83
EC
1.350
1.35 (EC), 6.07 (α),

EC = 100, α < 0.2

Bk-248m1

23.7

β−
0.870
β− = 70, EC = 30,

α < 0.001, 0,87 (β−),

0.717 (EC), 5.803 (α)

Bk-249

320.00
β−
0.125

Bk-250

3.22

β−
1.780

Bk-251
55.60

β−
1.093
β⁻ ≈ 100, α ≈ 1 · 10⁻⁵

Californium

Cf-241
3.78

EC
3.300
EC ≈ 75, α ≈ 25, 3.3 (EC),

7.66 (α)

Cf-242
3.49

α
7.516
α = 65, SF < 1.4 · 10⁻²

Cf-243
10.70

EC
2.220
EC ≈ 86, α ≈ 14, 2.22 (EC),

7.39 (α)

Cf-244
19.40
0.32

α
7.329

Cf-245
45.00
0.75

EC
1.569
EC = 64, α = 36, 1.569

(EC), 7.256 (α)

Cf-246

35.70
1.49
α
6.862
α ≈100, SF = 2, 3 · 10-4,

EC < 5 · 10-4

Cf-247

3.11

EC
0.646
EC ≈ 100, α = 0.035

Cf-248

333.50
α
6.361

Cf-253

17.81
β−
0.285

Cf-254

60.50
SF
5.926

Cf-255
85.00

β−
0.700

Cf-256
12.30

α
5.600
SF = 100, β⁻ < 1,

α ≈ 1 · 10⁻⁶

Einsteinium

Es-246
7.70

EC
3.880
EC = 90.1, α = 9.9,

ECSF = 0.003

Es-247
4.55

EC
2.480
2.48 (EC), 7.49 (α),

EC ≈ 93, α ≈ 7

Es-248
27.00
0.45

EC

Es-249
102.00
1.70

EC
1.450

Es-250

8.60

EC
2.100

Es-250m1
132.00
2.2

EC
2.100
2.10 (EC), 6.88 (α),

EC ≈ 100, α < 1

Es-251

33.00
1.38
EC
0.376

Es-253

20.47
α
6.739

Es-254m1

39.30
1.64
α, β−

Es-254

275.70
α
6.618

Es-255

39.80
β−
0.288

Es-256
25.40

β−
1.67

Es-256m1
456.00
7.6

β−
1.67
β⁻ ≈ 100, SF = 0.002

Es-257

7.8

Fermium

FM-249
2.60

EC
2.440
EC ≈ 85, α ≈ 15, 2.44 (EC),

7.81 (α)

Fm-250
30.00
0.50

α
7.557
7.557 (α), 0.8 (EC), α > 90,

EC < 10, SF = 0.0069

Fm-251

5.30

EC
1.474
1.474 (EC), 7.425 (α),

EC = 98.20, α = 1.80

Fm-252

22.70

α
7.425

Fm-253

72.00
3.00
EC
0.333
0.333 (EC), 7.197 (α),

EC = 88, α = 12

Fm-254

3.24

α
7.307
α ≈ 100, SF = 0.0592

Fm-255

20.07

α
7.241

Fm-256
157.60
2.60

α
7.027
SF = 91.9, α = 8.1

Fm-257

100.50
α
6.864

Mendelevium

Md-251
4.00

EC
3.070
3.07 (EC), 8.02 (α),

EC > 90, α < 10

Md-252
2.30

EC
3.89
EC > 50, α< 50

Md-253
6.00

ECβ+
1.96

Md-254
10.00

EC
2.68
EC < 100

Md-254m1
28.00

EC

EC < 100

Md-255
27.00

EC
1.043
1.043 (EC), 7.907 (α),

EC = 92, α = 8, SF < 1.4

Md-256
78.10

EC
2.130
2.13 (EC), 7.897 (α), EC = 90.7,

α = 9.3, SF < 2.8

Md-257

5.52

EC
0.406
0.406 (EC), 7.271 (α),

EC = 85, α = 15, SF < 1

Md-258

51.50
α
7.241
7.271(α), 1.23 (EC), α = 100,

SF < 0.003, β− < 0.003,

EC < 0.003

Md-258m1

57.00

EC
1.230
EC > 70, SF < 30, a < 1.2,

β− < 30

Md-259
96.00
1.60

α
7.100
SF > 73, α < 25, β− < 10,

7.0 (α), 1.0 (β−)

Md-260

27.80
α
7.000
SF > 73, α < 25, β⁻ < 10

Nobelium

No-255
3.10

α
8.445
α = 61.4, EC = 38.w6

No-259
58.00

α
7.910
α = 100, EC = 25, SF < 10

Lawrencium

Lr-261
39.00

SF

SF < 100

Lr-262
216.00

EC
2.1
EC > 10, SF < 10

Rutherfordium

Rf-263
15.00

SF

Seaborgium

Sg-271
2.40

α, SF

α > 50, SF < 50

Hassium

Hs-278
11.00

SF

Meitnerium

Mt-278
30.00

α
9.1

Roentgenium

Rg-282
4.00

α, SF
9.4

Nithonium

Nh-285
2.00

α, SF
10

Nh-286
5.00

α
9.7

Nh-287
20.00

α, SF
9.3

In an embodiment of the present invention, the radionuclide is used for diagnosis. Preferably, the radioactive isotope is selected from the group, but not limited to, comprising ⁴³Sc, ⁴⁴c, ⁵¹Mn, ⁵²Mn, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁸⁹Zr, ^94mTc, ^99mTc, ¹¹¹In, ¹⁵²Tb, ¹⁵⁵Tb, ¹⁷⁷Lu, ²⁰¹Tl, ²⁰³Pb, ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I. More preferably, the radionuclide is selected from the group comprising ⁴³Sc, ⁴⁴Sc, ⁶⁴Cu, ⁶⁷Ga, ⁶⁸Ga, ⁸⁶Y, ⁸⁹Zr, ^99mTc, ¹¹¹In, ¹⁵²Tb, ¹⁵⁵Tb, ²⁰³Pb, ¹⁸F, ⁷⁶Br, ⁷⁷Br, ¹²³I, ¹²⁴I, ¹²⁵I. Even more preferably, the radionuclide is selected from the group comprising ⁶⁴Cu, ⁶⁸Ga, ⁸⁹Zr, ^99mTc ¹¹¹In, ¹⁸F, ¹²³I, and ¹²⁴I. It will however, also be acknowledged by a person skilled in the art that the use of said radionuclide is not limited to diagnostic purposes, but encompasses their use in therapy and theragnostics when conjugated to the compound of the invention.

In an embodiment of the present invention, the radionuclide is used for therapy. Preferably, the radioactive isotope is selected from the group comprising ⁴⁷Sc, ⁶⁷Cu, ⁸⁹Sr, ⁹⁰Y, ¹¹¹In, ¹⁵³Sm, ¹⁴⁹Tb ¹⁶¹Tb, ¹⁷⁷Lu, ¹⁸⁶Re, ¹⁸⁸Re, ²¹²Pb ²¹³Bi, ²²³Ra, ²²⁵Ac, ²²⁶Th, ²²⁷Th, ¹³¹I, ²¹¹At. More preferably, the radioactive isotope is selected from the group comprising ⁴⁷Sc, ⁶⁷Cu, ⁹⁰Y, ¹⁷⁷Lu, ¹⁸⁸Re, ²¹²Pb, ²¹³Bi, ²²⁵Ac, ²²⁷Th, ¹³¹I, ²¹¹At. Even more preferably, the radionuclide is selected from the group comprising ⁹⁰Y, ¹⁷⁷Lu, ²²⁵Ac, ²²⁷Th, ¹³¹I and ²¹¹At. It will, however, also be acknowledged by a person skilled in the art that the use of said radionuclide is not limited to therapeutic purposes, but encompasses their use in diagnostic and theragnostics when conjugated to the compound of the invention.

In an embodiment the compound of the invention is present as a pharmaceutically acceptable salt.

A “pharmaceutically acceptable salt” of the compound of the present invention is preferably an acid salt or a base salt that is generally considered in the art to be suitable for use in contact with the tissues of human beings or animals without excessive toxicity or carcinogenicity, and preferably without irritation, allergic response, or other problem or complication. Such salts include mineral and organic acid salts of basic residues such as amines, as well as alkali or organic salts of acidic residues such as carboxylic acids. Compounds of the invention are capable of forming internal salts which are also pharmaceutically acceptable salts.

Suitable pharmaceutically acceptable salts include, but are not limited to, salts of acids such as hydrochloric, phosphoric, hydrobromic, malic, glycolic, fumaric, sulfuric, sulfamic, sulfanilic, formic, toluenesulfonic, methanesulfonic, benzene sulfonic, ethane disulfonic, 2-hydroxyethylsulfonic, nitric, benzoic, 2-acetoxybenzoic, citric, tartaric, lactic, stearic, salicylic, glutamic, ascorbic, pamoic, succinic, fumaric, maleic, propionic, hydroxymaleic, hydroiodic, phenylacetic, alkanoic such as acetic, HOOC—(CH₂)_n—COOH where n is any integer from 0 to 4, i.e., 0, 1, 2, 3, or 4, and the like. 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 further pharmaceutically acceptable salts for the compounds provided herein. In general, a pharmaceutically acceptable acid or base salt can be synthesized from a parent compound that contains a basic or acidic moiety by any conventional chemical method. Briefly, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. Generally, the use of non-aqueous media, such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile, is preferred.

A “pharmaceutically acceptable solvate” of the compound of the invention is preferably a solvate of the compound of the invention formed by association of one or more solvent molecules to one or more molecules of a compound of the invention. Preferably, the solvent is one which is generally considered in the art to be suitable for use in contact with the tissues of human beings or animals without excessive toxicity or carcinogenicity, and preferably without irritation, allergic response, or other problem or complication. Such solvent includes an organic solvent such as alcohols, ethers, esters and amines.

A “hydrate” of the compound of the invention is formed by association of one or more water molecules to one or more molecules of a compound of the invention. Such hydrate includes but is not limited to a hemi-hydrate, mono-hydrate, dihydrate, trihydrate and tetrahydrate. Independent of the hydrate composition all hydrates are generally considered as pharmaceutically acceptable.

The compound of the invention has a high binding affinity to FAP and a high inhibitory activity on FAP. Because of this high binding affinity, the compound of the invention is effective as, useful as and/or suitable as a targeting agent and, if conjugated to another moiety, as a targeting moiety. As preferably used herein a targeting agent is an agent which interacts with the target molecule which is in the instant case said FAP. In terms of cells and tissues thus targeted by the compound of the invention any cell and tissue, respectively, expressing said FAP is or may be targeted.

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

The identity between two nucleic acid molecules can be determined as known to the person skilled in the art. More specifically, a sequence comparison algorithm may be used for calculating the percent sequence homology for the test sequence(s) relative to the reference sequence, based on the designated program parameters. The test sequence is preferably the sequence or protein or polypeptide which is said to be identical or to be tested whether it is identical, and if so, to what extent, to a different protein or polypeptide, whereby such different protein or polypeptide is also referred to as the reference sequence and is preferably the protein or polypeptide of wild type, more preferably the human FAP of SEQ ID NO: 1.

Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman (Smith, et al., Advances in Applied Mathematics, 1981, 2: 482), by the homology alignment algorithm of 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: 2444), 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.

One example of an algorithm that is suitable for determining percent sequence identity is the algorithm used in the basic local alignment search tool (hereinafter “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 “NCBI”). The default parameters used in determining sequence identity using the software available from NCBI, e.g., 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).

It is within the present invention that the compound of the invention is used or is for use in a method for the treatment of a disease as disclosed herein. Such method, preferably, comprises the step of administering to a subject in need thereof a therapeutically effective amount of the compound of the invention. Such method includes, but is not limited to, curative or adjuvant cancer treatment. It is used as palliative treatment where cure is not possible and the aim is for local disease control or symptomatic relief or as therapeutic treatment where the therapy has survival benefit and it can be curative.

The method for the treatment of a disease as disclosed herein includes the treatment of the disease disclosed herein, including tumors and cancer, and may be used either as the primary therapy or as second, third, fourth or last line therapy. It is also within the present invention to combine the compound of the invention with further therapeutic approaches. It is well known to the person skilled in the art that the precise treatment intent including curative, adjuvant, neoadjuvant, therapeutic, or palliative treatment intent will depend on the tumor type, location, and stage, as well as the general health of the patient.

In an embodiment of the present invention, the disease is selected from the group comprising neoplasm nos, neoplasm benign, neoplasm uncertain whether benign or malignant, neoplasm malignant, neoplasm metastatic, neoplasm malignant uncertain whether primary or metastatic, tumor cells benign, tumor cells uncertain whether benign or malignant, tumor cells malignant, malignant tumor small cell type, malignant tumor giant cell type, malignant tumor fusiform cell type, epithelial neoplasms nos, epithelial tumor benign, carcinoma in situ nos, carcinoma nos, carcinoma metastatic nos, carcinomatosis, epithelioma benign, epithelioma malignant, large cell carcinoma nos, carcinoma undifferentiated type nos, carcinoma anaplastic type nos, pleomorphic carcinoma, giant cell and spindle cell carcinoma, giant cell carcinoma, spindle cell carcinoma, pseudosarcomatous carcinoma, polygonal cell carcinoma, spheroidal cell carcinoma, tumorlet, small cell carcinoma nos, oat cell carcinoma, small cell carcinoma, fusiform cell type, papillary and squamous cell neoplasms, papilloma nos, papillary carcinoma in situ, papillary carcinoma nos, verrucous papilloma, verrucous carcinoma nos, squamous cell papilloma, papillary squamous cell carcinoma, inverted papilloma, papillomatosis nos, squamous cell carcinoma in situ nos, squamous cell carcinoma nos, squamous cell carcinoma metastatic nos, squamous cell carcinoma, keratinizing type nos, squamous cell carcinoma large cell nonkeratinizing type, squamous cell carcinoma small cell nonkeratinizing type, squamous cell carcinoma spindle cell type, adenoid squamous cell carcinoma, squamous cell carcinoma in situ with questionable stromal invasion, squamous cell carcinoma microinvasive, queyrat's erythroplasia, bowen's disease, lymphoepithelial carcinoma, basal cell neoplasms, basal cell tumor, basal cell carcinoma nos, multicentric basal cell carcinoma, basal cell carcinoma, morphea type, basal cell carcinoma fibroepithelial type, basosquamous carcinoma, metatypical carcinoma, intraepidermal epithelioma of jadassohn, trichoepithelioma, trichofolliculoma, tricholemmoma, pilomatrixoma, transitional cell papillomas and carcinomas, transitional cell papilloma nos, urothelial papilloma, transitional cell carcinoma in situ, transitional cell carcinoma nos, schneiderian papilloma, transitional cell papilloma, inverted type, schneiderian carcinoma, transitional cell carcinoma spindle cell type, basaloid carcinoma, cloacogenic carcinoma, papillary transitional cell carcinoma, adenomas and adenocarcinomas, adenoma nos, bronchial adenoma nos, adenocarcinoma in situ, adenocarcinoma nos, adenocarcinoma metastatic nos, scirrhous adenocarcinoma, linitis plastica, superficial spreading adenocarcinoma, adenocarcinoma intestinal type, carcinoma diffuse type, monomorphic adenoma, basal cell adenoma, islet cell adenoma, islet cell carcinoma, insulinoma nos, insulinoma malignant, glucagonoma nos, glucagonoma malignant, gastrinoma nos, gastrinoma malignant, mixed islet cell and exocrine adenocarcinoma, bile duct adenoma, cholangiocarcinoma, bile duct cystadenoma, bile duct cystadenocarcinoma, liver cell adenoma, hepatocellular carcinoma nos, hepatocholangioma benign, combined hepatocellular carcinoma and cholangiocarcinoma, trabecular adenoma, trabecular adenocarcinoma, embryonal adenoma, eccrine dermal cylindroma, adenoid cystic carcinoma, cribriform carcinoma, adenomatous polyp nos, adenocarcinoma in adenomatous polyp, tubular adenoma nos, tubular adenocarcinoma, adenomatous polyposis coli, adenocarcinoma in adenomatous polyposis coli, multiple adenomatous polyps, solid carcinoma nos, carcinoma simplex, carcinoid tumor nos, carcinoid tumor malignant, carcinoid tumor argentaffin nos, carcinoid tumor argentaffin malignant, carcinoid tumor nonargentaffin nos, carcinoid tumor nonargentaffin malignant, mucocarcinoid tumor malignant, composite carcinoid, pulmonary adenomatosis, bronchiolo-alveolar adenocarcinoma, alveolar adenoma, alveolar adenocarcinoma, papillary adenoma nos, papillary adenocarcinoma nos, villous adenoma nos, adenocarcinoma in villous adenoma, villous adenocarcinoma, tubulovillous adenoma, chromophobe adenoma, chromophobe carcinoma, acidophil adenoma, acidophil carcinoma, mixed acidophil-basophil adenoma, mixed acidophil-basophil carcinoma, oxyphilic adenoma, oxyphilic adenocarcinoma, basophil adenoma, basophil carcinoma, clear cell adenoma, clear cell adenocarcinoma nos, hypemephroid tumor, renal cell carcinoma, clear cell adenofibroma, granular cell carcinoma, chief cell adenoma, water-clear cell adenoma, water-clear cell adenocarcinoma, mixed cell adenoma, mixed cell adenocarcinoma, lipoadenoma, follicular adenoma, follicular adenocarcinoma nos, follicular adenocarcinoma well differentiated type, follicular adenocarcinoma trabecular type, microfollicular adenoma, macrofollicular adenoma, papillary and follicular adenocarcinoma, nonencapsulated sclerosing carcinoma, multiple endocrine adenomas, juxtaglomerular tumor, adrenal cortical adenoma nos, adrenal cortical carcinoma, adrenal cortical adenoma compact cell type, adrenal cortical adenoma heavily pigmented variant, adrenal cortical adenoma clear cell type, adrenal cortical adenoma glomerulosa cell type, adrenal cortical adenoma mixed cell type, endometrioid adenoma nos, endometrioid adenoma, borderline malignancy, endometrioid carcinoma, endometrioid adenofibroma nos, endometrioid adenofibroma borderline malignancy, endometrioid adenofibroma malignant, adnexal and skin appendage neoplasms, skin appendage adenoma, skin appendage carcinoma, sweat gland adenoma, sweat gland tumor nos, sweat gland adenocarcinoma, apocrine adenoma, apocrine adenocarcinoma, eccrine acrospiroma, eccrine spiradenoma, hidrocystoma, papillary hydradenoma, papillary syringadenoma, syringoma nos, sebaceous adenoma, sebaceous adenocarcinoma, ceruminous adenoma, ceruminous adenocarcinoma, mucoepidermoid neoplasms, mucoepidermoid tumor, mucoepidermoid carcinoma cystic, mucinous, and serous neoplasms, cystadenoma nos, cystadenocarcinoma nos, serous cystadenoma nos, serous cystadenoma borderline malignancy, serous cystadenocarcinoma nos, papillary cystadenoma nos, papillary cystadenoma borderline malignancy, papillary cystadenocarcinoma nos, papillary serous cystadenoma nos, papillary serous cystadenoma borderline malignancy, papillary serous cystadenocarcinoma, serous surface papilloma nos, serous surface papilloma borderline malignancy, serous surface papillary carcinoma, mucinous cystadenoma nos, mucinous cystadenoma borderline malignancy, mucinous cystadenocarcinoma nos, papillary mucinous cystadenoma nos, papillary mucinous cystadenoma borderline malignancy, papillary mucinous cystadenocarcinoma, mucinous adenoma, mucinous adenocarcinoma, pseudomyxoma peritonei, mucin-producing adenocarcinoma, signet ring cell carcinoma, metastatic signet ring cell carcinoma, ductal, lobular, and medullary neoplasms, intraductal carcinoma noninfiltrating nos, infiltrating duct carcinoma, comedocarcinoma, noninfiltrating, comedocarcinoma nos, juvenile carcinoma of the breast, intraductal papilloma, noninfiltrating intraductal papillary adenocarcinoma, intracystic papillary adenoma, noninfiltrating intracystic carcinoma, intraductal papillomatosis nos, subareolar duct papillomatosis, medullary carcinoma nos, medullary carcinoma with amyloid stroma, medullary carcinoma with lymphoid stroma, lobular carcinoma in situ, lobular carcinoma nos, infiltrating ductular carcinoma, inflammatory carcinoma, paget's disease mammary, paget's disease and infiltrating duct carcinoma of breast, paget's disease extramammary, acinar cell neoplasms, acinar cell adenoma, acinar cell tumor, acinar cell carcinoma, complex epithelial neoplasms, adenosquamous carcinoma, adenolymphoma, adenocarcinoma with squamous metaplasia, adenocarcinoma with cartilaginous and osseous metaplasia, adenocarcinoma with spindle cell metaplasia, adenocarcinoma with apocrine metaplasia, thymoma benign, thymoma malignant, specialized gonadal neoplasms, sex cord-stromal tumor, thecoma nos, theca cell carcinoma, luteoma nos, granulosa cell tumor nos, granulosa cell tumor malignant, granulosa cell-theca cell tumor, androblastoma benign, androblastoma nos, androblastoma malignant, sertoli-leydig cell tumor, gynandroblastoma, tubular androblastoma nos, sertoli cell carcinoma, tubular androblastoma with lipid storage, leydig cell tumor benign, leydig cell tumor nos, leydig cell tumor malignant, hilar cell tumor, lipid cell tumor of ovary, adrenal rest tumor, paragangliomas and glomus tumors, paraganglioma nos, paraganglioma malignant, sympathetic paraganglioma, parasympathetic paraganglioma, glomus jugulare tumor, aortic body tumor, carotid body tumor, extra-adrenal paraganglioma nos, extra-adrenal paraganglioma, malignant, pheochromocytoma nos, pheochromocytoma malignant, glomangiosarcoma, glomus tumor, glomangioma, nevi and melanomas, pigmented nevus nos, malignant melanoma nos, nodular melanoma, balloon cell nevus, balloon cell melanoma, halo nevus, fibrous papule of the nose, neuronevus, magnocellular nevus, nonpigmented nevus, amelanotic melanoma, junctional nevus, malignant melanoma in junctional nevus, precancerous melanosis nos, malignant melanoma in precancerous melanosis, hutchinson's melanotic freckle, malignant melanoma in hutchinson's melanotic freckle, superficial spreading melanoma, intradermal nevus, compound nevus, giant pigmented nevus, malignant melanoma in giant pigmented nevus, epithelioid and spindle cell nevus, epithelioid cell melanoma, spindle cell melanoma nos, spindle cell melanoma type a, spindle cell melanoma type b, mixed epithelioid and spindle cell melanoma, blue nevus nos, blue nevus malignant, cellular blue nevus, soft tissue tumors and sarcomas nos, soft tissue tumor, benign, sarcoma nos, sarcomatosis nos, spindle cell sarcoma, giant cell sarcoma, small cell sarcoma, epithelioid cell sarcoma, fibromatous neoplasms, fibroma nos, fibrosarcoma nos, fibromyxoma, fibromyxosarcoma, periosteal fibroma, periosteal fibrosarcoma, fascial fibroma, fascial fibrosarcoma, infantile fibrosarcoma, elastofibroma, aggressive fibromatosis, abdominal fibromatosis, desmoplastic fibroma, fibrous histiocytoma nos, atypical fibrous histiocytoma, fibrous histiocytoma malignant, fibroxanthoma nos, atypical fibroxanthoma, fibroxanthoma malignant, dermatofibroma nos, dermatofibroma protuberans, dermatofibrosarcoma nos, myxomatous neoplasms, myxoma nos, myxosarcoma, lipomatous neoplasms, lipoma nos, liposarcoma nos, fibrolipoma, liposarcoma well differentiated type, fibromyxolipoma, myxoid liposarcoma, round cell liposarcoma, pleomorphic liposarcoma, mixed type liposarcoma, intramuscular lipoma, spindle cell lipoma, angiomyolipoma, angiomyoliposarcoma, angiolipoma nos, angiolipoma infiltrating, myelolipoma, hibemoma, lipoblastomatosis, myomatous neoplasms, leiomyoma nos, intravascular leiomyomatosis, leiomyosarcoma nos, epithelioid leiomyoma, epithelioid leiomyosarcoma, cellular leiomyoma, bizarre leiomyoma, angiomyoma, angiomyosarcoma, myoma, myosarcoma, rhabdomyoma nos, rhabdomyosarcoma nos, pleomorphic rhabdomyosarcoma, mixed type rhabdomyosarcoma, fetal rhabdomyoma, adult rhabdomyoma, embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma, complex mixed and stromal neoplasms, endometrial stromal sarcoma, endolymphatic stromal myosis, adenomyoma, pleomorphic adenoma, mixed tumor, malignant nos, mullerian mixed tumor, mesodermal mixed tumor, mesoblastic nephroma, nephroblastoma nos, epithelial nephroblastoma, mesenchymal nephroblastoma, hepatoblastoma, carcinosarcoma nos, carcinosarcoma embryonal type, myoepithelioma, mesenchymoma benign, mesenchymoma nos, mesenchymoma malignant, embryonal sarcoma, fibroepithelial neoplasms, brenner tumor nos, brenner tumor, borderline malignancy, brenner tumor malignant, fibroadenoma nos, intracanalicular fibroadenoma nos, pericanalicular fibroadenoma, adenofibroma nos, serous adenofibroma, mucinous adenofibroma, cellular intracanalicular fibroadenoma, cystosarcoma phyllodes nos, cystosarcoma phyllodes malignant, juvenile fibroadenoma, synovial neoplasms, synovioma benign, synovial sarcoma nos, synovial sarcoma spindle cell type, synovial sarcoma, epithelioid cell type, synovial sarcoma, biphasic type, clear cell sarcoma of tendons and aponeuroses, mesothelial neoplasms, mesothelioma benign, mesothelioma malignant, fibrous mesothelioma benign, fibrous mesothelioma malignant, epithelioid mesothelioma benign, epithelioid mesothelioma malignant, mesothelioma biphasic type benign, mesothelioma biphasic type malignant, adenomatoid tumor nos, germ cell neoplasms, dysgerminoma, seminoma nos, seminoma anaplastic type, spermatocytic seminoma, germinoma, embryonal carcinoma nos, endodermal sinus tumor, polyembryoma, gonadoblastoma, teratoma benign, teratoma nos, teratoma malignant nos, teratocarcinoma, malignant teratoma, undifferentiated type, malignant teratoma, intermediate type, dermoid cyst, dermoid cyst with malignant transformation, struma ovarii nos, struma ovarii malignant, strumal carcinoid, trophoblastic neoplasms, hydatidiform mole nos, invasive hydatidiform mole, choriocarcinoma, choriocarcinoma combined with teratoma, malignant teratoma trophoblastic, mesonephromas, mesonephroma benign, mesonephric tumor, mesonephroma malignant, endosalpingioma, blood vessel tumors, hemangioma nos, hemangiosarcoma, cavernous hemangioma, venous hemangioma, racemose hemangioma, kupffer cell sarcoma, hemangioendothelioma benign, hemangioendothelioma nos, hemangioendothelioma malignant, capillary hemangioma, intramuscular hemangioma, kaposi's sarcoma, angiokeratoma, verrucous keratotic hemangioma, hemangiopericytoma benign, hemangiopericytoma nos, hemangiopericytoma malignant, angiofibroma nos, hemangioblastoma, lymphatic vessel tumors, lymphangioma nos, lymphangiosarcoma, capillary lymphangioma, cavernous lymphangioma, cystic lymphangioma, lymphangiomyoma, lymphangiomyomatosis, hemolymphangioma, osteomas and osteosarcomas, osteoma nos, osteosarcoma nos, chondroblastic osteosarcoma, fibroblastic osteosarcoma, telangiectatic osteosarcoma, osteosarcoma in paget's disease of bone, juxtacortical osteosarcoma, osteoid osteoma nos, osteoblastoma, chondromatous neoplasms, osteochondroma, osteochondromatosis nos, chondroma nos, chondromatosis nos, chondrosarcoma nos, juxtacortical chondroma, juxtacortical chondrosarcoma, chondroblastoma nos, chondroblastoma malignant, mesenchymal chondrosarcoma, chondromyxoid fibroma, giant cell tumors, giant cell tumor of bone nos, giant cell tumor of bone malignant, giant cell tumor of soft parts nos, malignant giant cell tumor of soft parts, miscellaneous bone tumors, ewing's sarcoma, adamantinoma of long bones, ossifying fibroma, odontogenic tumors, odontogenic tumor benign, odontogenic tumor nos, odontogenic tumor malignant, dentinoma, cementoma nos, cementoblastoma benign, cementifying fibroma, gigantiform cementoma, odontoma nos, compound odontoma, complex odontoma, ameloblastic fibro-odontoma, ameloblastic odontosarcoma, adenomatoid odontogenic tumor, calcifying odontogenic cyst, ameloblastoma nos, ameloblastoma malignant, odontoameloblastoma, squamous odontogenic tumor, odontogenic myxoma, odontogenic fibroma nos, ameloblastic fibroma, ameloblastic fibrosarcoma, calcifying epithelial odontogenic tumor, miscellaneous tumors, craniopharyngioma, pinealoma, pineocytoma, pineoblastoma, melanotic neuroectodermal tumor, chordoma, gliomas, glioma malignant, gliomatosis cerebri, mixed glioma, subependymal glioma, subependymal giant cell astrocytoma, choroid plexus papilloma nos, choroid plexus papilloma malignant, ependymoma nos, ependymoma anaplastic type, papillary ependymoma, myxopapillary ependymoma, astrocytoma nos, astrocytoma, anaplastic type, protoplasmic astrocytoma, gemistocytic astrocytoma, fibrillary astrocytoma, pilocytic astrocytoma, spongioblastoma nos, spongioblastoma polare, astroblastoma, glioblastoma nos, giant cell glioblastoma, glioblastoma with sarcomatous component, primitive polar spongioblastoma, oligodendroglioma nos, oligodendroglioma, anaplastic type, oligodendroblastoma, medulloblastoma nos, desmoplastic medulloblastoma, medullomyoblastoma, cerebellar sarcoma nos, monstrocellular sarcoma, neuroepitheliomatous neoplasms, ganglioneuroma, ganglioneuroblastoma, ganglioneuromatosis, neuroblastoma nos, medulloepithelioma nos, teratoid medulloepithelioma, neuroepithelioma nos, spongioneuroblastoma, ganglioglioma, neurocytoma, pacinian tumor, retinoblastoma nos, retinoblastoma differentiated type, retinoblastoma undifferentiated type, olfactory neurogenic tumor, esthesioneurocytoma, esthesioneuroblastoma, esthesioneuroepithelioma, meningiomas, meningioma nos, meningiomatosis nos, meningioma malignant, meningotheliomatous meningioma, fibrous meningioma, psammomatous meningioma, angiomatous meningioma, hemangioblastic meningioma, hemangiopericytic meningioma, transitional meningioma, papillary meningioma, meningeal sarcomatosis, nerve sheath tumor, neurofibroma nos, neurofibromatosis nos, neurofibrosarcoma, melanotic neurofibroma, plexiform neurofibroma, neurilemmoma nos, neurinomatosis, neurilemmoma malignant, neuroma nos, granular cell tumors and alveolar soft part sarcoma, granular cell tumor nos, granular cell tumor, malignant, alveolar soft part sarcoma, lymphomas nos or diffuse, lymphomatous tumor benign, malignant lymphoma nos, malignant lymphoma non hodgkin's type, malignant lymphoma, undifferentiated cell type nos, malignant lymphoma stem cell type, malignant lymphoma convoluted cell type nos, lymphosarcoma nos, malignant lymphoma lymphoplasmacytoid type, malignant lymphoma immunoblastic type, malignant lymphoma mixed lymphocytic-histiocytic nos, malignant lymphoma centroblastic-centrocytic diffuse, malignant lymphoma follicular center cell nos, malignant lymphoma lymphocytic well differentiated nos, malignant lymphoma lymphocytic intermediate differentiation nos, malignant lymphoma centrocytic malignant lymphoma follicular center cell, cleaved nos, malignant lymphoma lymphocytic poorly differentiated nos, prolymphocytic lymphosarcoma, malignant lymphoma centroblastic type nos, malignant lymphoma follicular center cell noncleaved nos, reticulosarcomas, reticulosarcoma nos, reticulosarcoma pleomorphic cell type, reticulosarcoma nodular, hodgkin's disease, hodgkin's disease nos, hodgkin's disease lymphocytic predominance, hodgkin's disease mixed cellularity, hodgkin's disease lymphocytic depletion nos, hodgkin's disease lymphocytic depletion diffuse fibrosis, hodgkin's disease lymphocytic depletion reticular type, hodgkin's disease nodular sclerosis nos, hodgkin's disease nodular sclerosis cellular phase, hodgkin's paragranuloma, hodgkin's granuloma, hodgkin's sarcoma, lymphomas nodular or follicular, malignant lymphoma nodular nos, malignant lymphoma mixed lymphocytic-histiocytic nodular, malignant lymphoma centroblastic-centrocytic follicular, malignant lymphoma lymphocytic well differentiated nodular, malignant lymphoma lymphocytic intermediate differentiation nodular, malignant lymphoma follicular center cell cleaved follicular, malignant lymphoma lymphocytic poorly differentiated nodular, malignant lymphoma centroblastic type follicular malignant lymphoma follicular center cell noncleaved follicular, mycosis fungoides, mycosis fungoides, sezary's disease, miscellaneous reticuloendothelial neoplasms, microglioma, malignant histiocytosis, histiocytic medullary reticulosis, letterer-siwe's disease, plasma cell tumors, plasma cell myeloma, plasma cell tumor, benign, plasmacytoma nos, plasma cell tumor malignant, mast cell tumors, mastocytoma nos, mast cell sarcoma, malignant mastocytosis, burkitt's tumor, burkitt's tumor, leukemias, leukemias nos, leukemia nos, acute leukemia nos, subacute leukemia nos, chronic leukemia nos, aleukemic leukemia nos, compound leukemias, compound leukemia, lymphoid leukemias, lymphoid leukemia nos, acute lymphoid leukemia, subacute lymphoid leukemia, chronic lymphoid leukemia, aleukemic lymphoid leukemia, prolymphocytic leukemia, plasma cell leukemias, plasma cell leukemia, erythroleukemias, erythroleukemia, acute erythremia, chronic erythremia, lymphosarcoma cell leukemias, lymphosarcoma cell leukemia, myeloid leukemias, myeloid leukemia nos, acute myeloid leukemia, subacute myeloid leukemia, chronic myeloid leukemia, aleukemic myeloid leukemia, neutrophilic leukemia, acute promyelocytic leukemia, basophilic leukemias, basophilic leukemia, eosinophilic leukemias, eosinophilic leukemia, monocytic leukemias, monocytic leukemia nos, acute monocytic leukemia, subacute monocytic leukemia, chronic monocytic leukemia, aleukemic monocytic leukemia, miscellaneous leukemias, mast cell leukemia, megakaryocytic leukemia, megakaryocytic myelosis, myeloid sarcoma, hairy cell leukemia, miscellaneous myeloproliferative and lymphoproliferative disorders, polycythemia vera, acute panmyelosis, chronic myeloproliferative disease, myelosclerosis with myeloid metaplasia, idiopathic thrombocythemia, chronic lymphoproliferative disease.

In an embodiment of the present invention, the disease is selected from the group comprising tumors of pancreas, pancreatic adenocarcinoma, tumors of head of pancreas, of body of pancreas, of tail of pancreas, of pancreatic duct, of islets of langerhans, neck of pancreas, tumor of prostate, prostate adenocarcinoma, prostate gland, neuroendocrine tumors, breast cancer, tumor of central portion of breast, upper inner quadrant of breast, lower inner quadrant of breast, upper outer quadrant of breast, lower outer quadrant of breast, axillary tail of breast, overlapping lesion of breast, juvenile carcinoma of the breast, tumors of parathyroid gland, myeloma, lung cancer, small cell lung cancer, non-small cell lung cancer, tumor of main bronchus, of upper lobe lung, of middle lobe lung, of lower lobe lung, colorectal carcinoma, tumor of ascending colon, of hepatic flexure of colon, of transverse colon, of splenic flexure of colon, of descending colon, of sigmoid colon, of overlapping lesion of colon, of small intestine, tumors of liver, liver cell adenoma, hepatocellular carcinoma, hepatocholangioma, ombined hepatocellular carcinoma and cholangiocarcinoma, hepatoblastoma, ovarian carcinoma, sarcoma, osteosarcoma, fibrosarcoma, gastrointestinal stroma tumors, gastrointestinal tract, gastric carcinoma, thyroid carcinoma, medullary thyroid carcinoma, thyroid gland, renal cell carcinoma, renal pelvis, tumors of bladder, bladder carcinoma, tumors of trigone bladder, of dome bladder, of lateral wall bladder, of posterior wall bladder, of ureteric orifice, of urachus, overlapping lesion of bladder, basal cell carcinoma, basal cell neoplasms, basal cell tumor, basal cell carcinoma, multicentric basal cell carcinoma, basaloid carcinoma, basal cell adenoma, squamous cell carcinoma, oral squamous cell carcinoma, squamous cell carcinoma of the larynx, cervical carcinoma, tumors of exocervix, of overlapping lesion of cervix uteri, of cervix uteri, of isthmus uteri, tumors of uterus, tumors of ovary, tumors of cervical esophagus, of thoracic esophagus, of abdominal esophagus, of upper third of esophagus, of esophagus middle third, of esophagus lower third, of overlapping lesion of esophagus, endometrial carcinoma, head and neck cancer, lymphoma, malignant mesothelioma, mesothelial neoplasms, mesothelioma, fibrous mesothelioma, fibrous mesothelioma, epithelioid mesothelioma, epithelioid mesothelioma, duodenal carcinoma, neuroendocrine tumors, neuroendocrine tumors of the lung, neuroendocrine tumors of the pancreas, neuroendocrine tumors of the foregut, neuroendocrine tumors of the midgut, neuroendocrine tumors of the hindgut, gastroenteropancreatic neuroendocrine tumors, neuroendocrine carcinomas, neuroendocrine tumors of the breast, neuroendocrine tumors of the ovaries, testicular cancer, thymic carcinoma, tumors of stomach, fundus stomach, body stomach, gastric antrum, pylorus, lesser curvature of stomach, greater curvature of stomach, overlapping lesion of stomach, paragangliomas, ganglioma, melanomas, malignant melanoma, nodular melanoma, amelanotic melanoma, superficial spreading melanoma, epithelioid cell melanoma, spindle cell melanoma, mixed epithelioid and spindle cell melanoma.

In a still further embodiment, the aforementioned indications may occur in organs and tissues selected from the group comprising external upper lip, external lower lip, external lip nos, upper lip mucosa, lower lip mucosa, mucosa lip nos, commissure lip, overlapping lesion of lip, base of tongue nos, dorsal surface tongue nos, border of tongue, ventral surface of tongue nos, anterior 2/3 of tongue nos, lingual tonsil, overlapping lesion of tongue, tongue nos, upper gum, lower gum, gum nos, anterior floor of mouth, lateral floor of mouth, overlapping lesion of floor of mouth, floor of mouth nos, hard palate, soft palate nos, uvula, overlapping lesion of palate, palate nos, cheek mucosa, vestibule of mouth, retromolar area, overlapping lesion of other and unspecified parts of mouth, mouth nos, parotid gland, submaxillary gland, sublingual gland, overlapping lesion of major salivary glands, major salivary gland nos, tonsillar fossa, tonsillar pillar, overlapping lesion of tonsil, tonsil nos, vallecula, anterior surface of epiglottis, lateral wall oropharynx, posterior wall oropharynx, branchial cleft, overlapping lesion of oropharynx, oropharynx nos, superior wall of nasopharynx, posterior wall nasopharynx, lateral wall nasopharynx, anterior wall nasopharynx, overlapping lesion of nasopharynx, nasopharynx nos, pyriform sinus, postcricoid region, hypopharyngeal aspect of aryepiglottic fold, posterior wall hypopharynx, overlapping lesion of hypopharynx, hypopharynx nos, pharynx nos, laryngopharynx, waldeyer's ring, overlapping lesion of lip oral cavity and pharynx, cervical esophagus, thoracic esophagus, abdominal esophagus, upper third of esophagus, middle third of esophagus, esophagus lower third, overlapping lesion of esophagus, esophagus nos, cardia nos, fundus stomach, body stomach, gastric antrum, pylorus, lesser curvature of stomach nos, greater curvature of stomach nos, overlapping lesion of stomach, stomach nos, duodenum, jejunum, ileum, meckel's diverticulum, overlapping lesion of small intestine, small intestine nos, cecum, appendix, ascending colon, hepatic flexure of colon, transverse colon, splenic flexure of colon, descending colon, sigmoid colon, overlapping lesion of colon, colon nos, rectosigmoid junction, rectum nos, anus nos, anal canal, cloacogenic zone, overlapping lesion of rectum anus and anal canal, liver, intrahepatic bile duct, gallbladder, extrahepatic bile duct, ampulla of vater, overlapping lesion of biliary tract, biliary tract nos, head of pancreas, body pancreas, tail pancreas, pancreatic duct, islets of langerhans, neck of pancreas, overlapping lesion of pancreas, pancreas nos, intestinal tract nos, overlapping lesion of digestive system, gastrointestinal tract nos, nasal cavity, middle ear, maxillary sinus, ethmoid sinus, frontal sinus, sphenoid sinus, overlapping lesion of accessory sinuses, accessory sinus nos, glottis, supraglottis, subglottis, laryngeal cartilage, overlapping lesion of larynx, larynx nos, trachea, main bronchus, upper lobe lung, middle lobe lung, lower lobe lung, overlapping lesion of lung, lung nos, thymus, heart, anterior mediastinum, posterior mediastinum, mediastinum nos, pleura nos, overlapping lesion of heart mediastinum and pleura, upper respiratory tract nos, overlapping lesion of respiratory system and intrathoracic organs, respiratory tract nos, upper limb long bones joints, upper limb short bones joints, lower limb long bones joints, lower limb short bones joints, overlapping lesion of bones joints and articular cartilage of limbs, bone limb nos, skull and facial bone, mandible, vertebral column, rib sternum clavicle, pelvic bone, overlapping lesion of bones joints and articular cartilage, bone nos, blood, bone marrow, spleen, reticuloendothelial system nos, hematopoietic system nos, skin lip nos, eyelid nos, external ear, skin face, skin scalp neck, skin trunk, skin limb upper, skin limb lower, peripheral nerve head neck, peripheral nerve shoulder arm, peripheral nerve leg, peripheral nerve thorax, peripheral nerve abdomen, peripheral nerve pelvis, peripheral nerve trunk, overlapping lesion of peripheral nerves and autonomic nervous system, autonomic nervous system nos, retroperitoneum, peritoneum, peritoneum nos, overlapping lesion of retroperitoneum and peritoneum, connective tissue head, connective tissue arm, connective tissue leg, connective tissue thorax, connective tissue abdomen, connective tissue pelvis, connective tissue trunk nos, overlapping lesion of connective subcutaneous and other soft tissues, connective tissue nos, nipple, central portion of breast, upper inner quadrant of breast, lower inner quadrant of breast, upper outer quadrant of breast, lower outer quadrant of breast, axillary tail of breast, overlapping lesion of breast, breast nos, labium majus, labium minus, clitoris, overlapping lesion of vulva, vulva nos, vagina nos, endocervix, exocervix, overlapping lesion of cervix uteri, cervix uteri, isthmus uteri, endometrium, myometrium, fundus uteri, overlapping lesion of corpus uteri, corpus uteri, uterus nos, ovary, fallopian tube, broad ligament, round ligament, parametrium, uterine adnexa, wolffian body, overlapping lesion of female genital organs, female genital tract nos, prepuce, glans penis, body penis, overlapping lesion of penis, penis nos, prostate gland, undescended testis, descended testis, testis nos, epididymis, spermatic cord, scrotum nos, tunica vaginalis, overlapping lesion of male genital organs, male genital organs nos, kidney nos, renal pelvis, ureter, trigone bladder, dome bladder, lateral wall bladder, posterior wall bladder, ureteric orifice, urachus, overlapping lesion of bladder, bladder nos, urethra, paraurethral gland, overlapping lesion of urinary organs, urinary system nos, conjunctiva, comea nos, retina, choroid, ciliary body, lacrimal gland, orbit nos, overlapping lesion of eye and adnexa, eye nos, cerebral meninges, spinal meninges, meninges nos, cerebrum, frontal lobe, temporal lobe, parietal lobe, occipital lobe, ventricle nos, cerebellum nos, brain stem, overlapping lesion of brain, brain nos, spinal cord, cauda equina, olfactory nerve, optic nerve, acoustic nerve, cranial nerve nos, overlapping lesion of brain and central nervous system, nervous system nos, thyroid gland, adrenal gland cortex, adrenal gland medulla, adrenal gland nos, parathyroid gland, pituitary gland, craniopharyngeal duct, pineal gland, carotid body, aortic body, overlapping lesion of endocrine glands and related structures, endocrine gland nos, head face or neck nos, thorax nos, abdomen nos, pelvis nos, upper limb nos, lower limb nos, other illdefined sites, overlapping lesion of ill-defined sites, lymph node face head neck, intrathoracic lymph node, intra-abdominal lymph nodes, lymph node axilla arm, lymph node inguinal region leg, lymph node pelvic, lymph nodes of multiple regions, lymph node nos, unknown primary site.

The subjects treated with the presently disclosed and claimed compounds may be treated in combination with other non-surgical anti-proliferative (e.g., anti-cancer) drug therapy. 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, a nucleic acid, or a protein) that suppresses cell growth and/or proliferation. In some embodiments, the cytostatic compound is directed towards the malignant cells of a tumor. In yet other embodiments, the cytostatic compound is one which inhibits the growth and/or proliferation of vascular smooth muscle cells or fibroblasts.

Suitable anti-proliferative drugs or cytostatic compounds to be used in combination with the presently disclosed and claimed compounds include anti-cancer drugs. Numerous anti-cancer drugs which may be used are well known and include, but are not limited to: Acivicin; Aclarubicin; Acodazole Hydrochloride; Acronine; Adozelesin; Aldesleukin; Altretamine; Ambomycin; Ametantrone Acetate; Aminoglutethimide; Amsacrine; Anastrozole; Anthramycin; Asparaginase; Asperlin; Azacitidine; Azetepa; Azotomycin; Batimastat; Benzodepa; Bicalutamide; Bisantrene Hydrochloride; Bisnafide Dimesylate; Bizelesin; Bleomycin Sulfate; Brequinar Sodium; Bropirimine; Busulfan; Cactinomycin; Calusterone; Caracemide; Carbetimer; Carboplatin; Carmustine; Carubicin Hydrochloride; Carzelesin; Cedefingol; Chlorambucil; Cirolemycin; Cisplatin; Cladribine; Crisnatol Mesylate; Cyclophosphamide; Cytarabine; Dacarbazine; Dactinomycin; Daunorubicin Hydrochloride; Decitabine; Dexormaplatin; Dezaguanine; Dezaguanine Mesylate; Diaziquone; Docetaxel; Doxorubicin; Doxorubicin Hydrochloride; Droloxifene; Droloxifene Citrate; Dromostanolone Propionate; Duazomycin; Edatrexate; Eflomithine Hydrochloride; Elsamitrucin; Enloplatin; Enpromate; Epipropidine; Epirubicin Hydrochloride; Erbulozole; Esorubicin Hydrochloride; Estramustine; Estramustine Phosphate Sodium; Etanidazole; Etoposide; Etoposide Phosphate; Etoprine; Fadrozole Hydrochloride; Fazarabine; Fenretinide; Floxuridine; Fludarabine Phosphate; Fluorouracil; Fluorocitabine; Fosquidone; Fostriecin Sodium; Gemcitabine; Gemcitabine Hydrochloride; Hydroxyurea; Idarubicin Hydrochloride; Ifosfamide; Ilmofosine; Interferon Alfa-2a; Interferon Alfa-2b; Interferon Alfa-n1; Interferon Alfa-n3; Interferon Beta-I a; Interferon Gamma-I b; 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; Mitocarcin; Mitocromin; Mitogillin; Mitomalcin; Mitomycin; Mitosper; Mitotane; Mitoxantrone Hydrochloride; Mycophenolic Acid; Niraparib; Nocodazole; Nogalamycin; Olaparib; Ormaplatin; Oxisuran; Paclitaxel; Pegaspargase; Peliomycin; Pentamustine; Peplomycin Sulfate; Perfosfamide; Pipobroman; Piposulfan; Piroxantrone Hydrochloride; Plicamycin; Plomestane; Porfimer Sodium; Porfiromycin; Prednimustine; Procarbazine Hydrochloride; Puromycin; Puromycin Hydrochloride; Pyrazofurin; Riboprine; Rogletimide; Rucaparib; Safingol; Safingol Hydrochloride; Semustine; Simtrazene; Sparfosate Sodium; Sparsomycin; Spirogermanium Hydrochloride; Spiromustine; Spiroplatin; Streptonigrin; Streptozocin; Sulofenur; Talazoparib; Talisomycin; Taxol; Taxotere; Tecogalan Sodium; Tegafur; Teloxantrone Hydrochloride; Temoporfin; Teniposide; Teroxirone; Testolactone; Thiamiprine; Thioguanine; Thiotepa; Tiazofurin; Tirapazamine; Topotecan Hydrochloride; Toremifene Citrate; Trestolone Acetate; Triciribine Phosphate; Trimetrexate; Trimetrexate Glucuronate; Tubulozole Hydrochloride; Uracil Mustard; Uredepa; Vapreotide; Velaparib; Verteporfin; Vinblastine Sulfate; Vincristine Sulfate; Vindesine; Vindesine Sulfate; Vinepidine Sulfate; Vinglycinate Sulfate; Vinleurosine Sulfate; Vinorelbine Tartrate; Vinrosidine Sulfate; Vinzolidine Sulfate; Vorozole; Zeniplatin; Zinostatin; and Zorubicin Hydrochloride.

Other anti-cancer drugs include, but are not limited to: 20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; acylfulvene; adecypenol; adozelesin; ALL-TK antagonists; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; anagrelide; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bisaziridinylspermine; bisnafide; bistratene A; breflate; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; dehydrodidemnin B; deslorelin; dexifosfamide; dexrazoxane; dexverapamil; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflomithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue; estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; flgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-I receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; irinotecan; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1-based therapy; mustard anti cancer compound; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; osaterone; oxaliplatin; oxaunomycin; paclitaxel analogues; paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum-triamine complex; porfimer sodium; porfiromycin; propyl bis-acridone; prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists; raltitrexed; ramosetron; ras famesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temozolomide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thalidomide; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; 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 system, erythrocyte gene therapy; velaresol; veramine; verdins; vinorelbine; vinxaltine; vitaxin; zanoterone; zilascorb; and zinostatin stimalamer.

The presently disclosed and claimed compounds can also be used in combination with any of the following treatments:

Therapy in combination with inhibitors of Poly(ADP-ribose) polymerases (PARP), a class of chemotherapeutic agents directed at targeting 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.

Therapy in combination with inhibitors of signaling pathways and mechanisms leading to repair of DNA single and double strand breaks as e.g. nuclear factor-kappaB signaling (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 kinases, checkpoint kinase 1 and 2, DNA-dependen protein kinase, and WEEl kinase (Pilie, et al., Nat Rev Clin Oncol, 2019, 16: 81).

Therapy in combination with an immunomodulator (Khalil, et al., Nat Rev Clin Oncol, 2016, 13: 394), a cancer vaccine (Hollingsworth, et al., NPJ Vaccines, 2019, 4: 7), an immune checkpoint inhibitor (e.g. PD-1, PD-L1, CTLA-4-inhibitor) (Wei, et al., Cancer Discov, 2018, 8: 1069), a Cyclin-D-Kinase 4/6 inhibitor (Goel, et al., Trends Cell Biol, 2018, 28: 911), an antibody being capable of binding to a tumor cell and/or metastases and being capable of inducing antibody-dependent cellular cytotoxicity (ADCC) (Kellner, et al., Transfus Med Hemother, 2017, 44: 327), a T cell- or NK cell engager (e.g. bispecific antibodies) (Yu, et al., J Cancer Res Clin Oncol, 2019, 145: 941), a cellular 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 incluce but are not limited to nivolumab, ipilimumab, pembrolizumab, atezolizumab, avelumab, durvalumab, and cemiplimab.

According to the present invention, the compounds may be administered prior to, concurrent with, or following other anti-cancer compounds. The administration schedule may involve administering the different agents in an alternating fashion. In other embodiments, the compounds may be delivered before and during, or 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 the other anti-proliferative treatment. In other embodiments, more than one anti-proliferative therapy may be administered to a subject. For example, the subject may receive the present compounds, in combination with both surgery and at least one other anti-proliferative compound. Alternatively, the compound may be administered in combination with more than one anti-cancer drug.

In an embodiment, the compounds of the present invention are used to detect cells and tissues overexpressing FAP, whereby such detection is achieved by conjugating a detectable label to the compounds of the invention, preferably a detectable radionuclide. In a preferred embodiment, the cells and tissues detected are diseased cells and tissues and/or are either a or the cause for the disease and/or the symptoms of the disease, or are part of the pathology underlying the disease. In a further preferred embodiment, the diseased cells and tissues are causing and/or are part of an oncology indication (e.g. neoplasms, tumors, and cancers) or a non-oncology indication (e.g. inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease).

In another embodiment, the compounds of the present invention are used to treat cells and tissues overexpressing FAP. In a preferred embodiment, the cells and tissues treated are diseased cells and tissues and/or are either a or the cause for the disease and/or the symptoms of the disease, or are part of the pathology underlying the disease. In a further preferred embodiment, the diseased cells and tissues are causing and/or are part of an oncology indication (e.g. neoplasms, tumors, and cancers) and the therapeutic activity is achieved by conjugating therapeutically active effector to the compounds of the present invention, preferably a therapeutically active radionuclide. In a further preferred embodiment, the diseased cells and tissues are causing and/or are part of a non-oncology indication (e.g. inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease) and the therapeutic activity is achieved by inhibition of the enzymatic activity of FAP.

In a further embodiment, particularly if the disease is a non-oncology disease or a non-oncology indication (e.g. inflammatory disease, cardiovascular disease, autoimmune disease, and fibrotic disease), the compounds of the present invention are administered in therapeutically effective amounts; preferably the compound of the present invention does not comprise a therapeutically active nuclide. An effective amount is a dosage of the compound sufficient to provide a therapeutically or medically desirable result or effect in the subject to which the compound is administered. The effective amount will vary with the particular condition being treated, the age and physical condition of the subject being treated, the severity of the condition, the duration of the treatment, the nature of the concurrent or combination therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health practitioner. For example, in connection with methods directed towards treating subjects having a condition characterized by abnormal cell proliferation, an effective amount to inhibit proliferation would be an amount sufficient to reduce or halt altogether the abnormal cell proliferation so as to slow or halt the development of or the progression of a cell mass such as, for example, a tumor. As used in the embodiments, “inhibit” embraces all of the foregoing.

In other embodiments, a therapeutically effective amount will be an amount necessary to extend the dormancy of micrometastases or to stabilize any residual primary tumor cells following surgical or drug therapy.

Generally, when using an unconjugated compound without a therapeutically active radionuclide, a therapeutically effective amount will vary with the subject's age, condition, and sex, 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. The dosage may be adjusted by the individual physician or veterinarian, particularly in the event of any complication. A therapeutically effective amount is typically, but not limited to, an amount in a range from 0.1 μg/kg to about 2000 mg/kg, or from 1.0 μg/kg to about 1000 mg/kg, or from about 0.1 mg/kg to about 500 mg/kg, or from about 1.0 mg/kg to about 100 mg/kg, in one or more dose administrations daily, 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 sub-doses for example administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In some embodiments, the compounds are 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 over a period of weeks, or months. In still other embodiments, the compound is delivered on alternate days. For example, the agent is delivered every two days, or every three days, or every four days, or every five days, or every six days, or every week, or every month.

In a preferred embodiment, the compound of the present invention is for use in the treatment and/or prevention of a disease, whereby such treatment is radionuclide therapy.

Preferably, radionuclide therapy makes use of or is based on different forms of radiation emitted by a radionuclide. Such radiation can, for example, be any one of radiation of photons, radiation of electrons including but not limited to β⁻-particles and Auger-electrons, radiation of protons, radiation of neutrons, radiation of positrons, radiation of α-particles or an ion beam. Depending on the kind of particle or radiation emitted by said radionuclide, radionuclide therapy can, for example, be distinguished as photon radionuclide therapy, electron radionuclide therapy, proton radionuclide therapy, neutron radionuclide therapy, positron radionuclide therapy, α-particle radionuclide therapy or ion beam radionuclide therapy. All of these forms of radionuclide therapy are encompassed by the present invention, and all of these forms of radionuclide therapy can be realized by the compound of the invention, preferably under the proviso that the radionuclide attached to the compound of the invention, more preferably as an effector, is providing for this kind of radiation.

Radionuclide therapy preferably works by damaging the DNA of cells. The damage is caused by a photon, electron, proton, neutron, positron, α-particle or ion beam directly or indirectly ionizing the atoms which make up the DNA chain. Indirect ionization happens as a result of the ionization of water, forming free radicals, notably hydroxyl radicals, which then damage the DNA.

In the most common forms of radionuclide therapy, most of the radiation effect is through free radicals. Because cells have mechanisms for repairing DNA damage, breaking the DNA on both strands proves to be the most significant technique in modifying cell characteristics.

Because cancer cells generally are undifferentiated and stem cell-like, they reproduce more, and have a diminished ability to repair sub-lethal damage compared to most healthy differentiated cells. The DNA damage is inherited through cell division, accumulating damage to the cancer cells, causing them to die or reproduce more slowly.

Oxygen is a potent radiosensitizer, increasing the effectiveness of a given dose of radiation by forming DNA-damaging free radicals. Therefore, use of high pressure oxygen tanks, blood substitutes that carry increased oxygen, hypoxic cell radiosensitizers such as misonidazole and metronidazole, and hypoxic cytotoxins, such as tirapazamine may be applied.

Other factors that are considered when selecting a radioactive dose include whether the patient is receiving chemotherapy, whether radiation therapy is being administered before or after surgery, and the degree of success of surgery.

The total radioactive dose may be fractionated, i.e. spread out over time in one or more treatments for several important reasons. Fractionation allows normal cells time to recover, while tumor cells are generally less efficient in repair between fractions. Fractionation also allows tumor cells that were in a relatively radio-resistant phase of the cell cycle during one treatment to cycle into a sensitive phase of the cycle before the next fraction is given. Similarly, tumor cells that were chronically or acutely hypoxic and, therefore, more radioresistant, may reoxygenate between fractions, improving the tumor cell kill.

It is generally known that different cancers respond differently to radiation therapy. The response of a cancer to radiation is described by its radiosensitivity. Highly radiosensitive cancer cells are rapidly killed by modest doses of radiation. These include leukemias, most lymphomas, and germ cell tumors.

It is important to distinguish radiosensitivity of a particular tumor, which to some extent is a laboratory measure, from “curability” of a cancer by an internally delivered radioactive dose in actual clinical practice. For example, leukemias are not generally curable with radiotherapy, because they are disseminated through the body. Lymphoma may be radically curable if it is localized to one area of the body. Similarly, many of the common, moderately radioresponsive tumors can be treated with curative doses of radioactivity if they are at an early stage. This applies, for example, to non-melanoma skin cancer, head and neck cancer, non-small cell lung cancer, cervical cancer, anal cancer, prostate cancer.

The response of a tumor to radiotherapy is also related to its size. For complex reasons, very large tumors respond less well to radiation than smaller tumors or microscopic disease. Various strategies are used to overcome this effect. The most common technique is surgical resection prior to radiotherapy. This is most commonly seen in the treatment of breast cancer with wide local excision or mastectomy followed by adjuvant radiotherapy. Another method is to shrink the tumor with neoadjuvant chemotherapy prior to radical radionuclide therapy. A third technique is to enhance the radiosensitivity of the cancer by giving certain drugs during a course of radiotherapy. Examples of radiosensiting drugs include, but are not limited to Cisplatin, Nimorazole, and Cetuximab.

Introperative radiotherapy is a special type of radiotherapy that is delivered immediately after surgical removal of the cancer. This method has been employed in breast cancer (TARGeted Introperative radio Therapy), brain tumors and rectal cancers.

Radionuclide therapy is in itself painless. Many low-dose palliative treatments cause minimal or no side effects. Treatment to higher doses may cause varying side effects during treatment (acute side effects), in the months or years following treatment (long-term side effects), or after re-treatment (cumulative side effects). The nature, severity, and longevity of side effects depends on the organs that receive the radiation, the treatment itself (type of radionuclide, dose, fractionation, concurrent chemotherapy), and the patient.

It is within the present inventions that the method for the treatment of a disease of the invention may realize each and any of the above strategies which are as such known in the art, and which insofar constitute further embodiments of the invention.

It is also within the present invention that the compound of the invention is used in a method for the diagnosis of a disease as disclosed herein. Such method, preferably, comprises the step of administering to a subject in need thereof a diagnostically effective amount of the compound of the invention.

In accordance with the present invention, an imaging method is selected from the group consisting of scintigraphy, Single Photon Emission Computed Tomography (SPECT) and Positron Emission Tomography (PET).

In a preferred embodiment of the present invention, a compound according to the present invention comprising a chelator from the N4 chelator family, more preferably chelating a Tc radionuclide, is particularly suitable for use in a method and procedure using SPECT. In an embodiment thereof, the chelator from the N4 chelator family is N4Ac.

In a preferred embodiment of the present invention, a compound according to the present invention comprising chelator NODAGA, more preferably chelating a Ga radionuclide is particularly suitable for use in a method and procedure using PET.

Scintigraphy is a form of diagnostic test or method used in nuclear medicine, wherein radiopharmaceuticals are internalized by cells, tissues and/or organs, preferably internalized in vivo, and radiation emitted by said internalized radiopharmaceuticals is captured by external detectors (gamma cameras) to form and display two-dimensional images. In contrast thereto, SPECT and PET forms and displays three-dimensional images. Because of this, SPECT and PET are classified as separate techniques to scintigraphy, although they also use gamma cameras to detect internal radiation. Scintigraphy is unlike a diagnostic X-ray where external radiation is passed through the body to form an image.

Single Photon Emission Tomography (SPECT) scans are a type of nuclear imaging technique using gamma rays. They are very similar to conventional nuclear medicine planar imaging using a gamma camera. Before the SPECT scan, the patient is injected with a radiolabeled chemical emitting gamma rays that can be detected by the scanner. A computer collects the information from the gamma camera and translates this into two-dimensional cross-sections. These cross-sections can be added back together to form a three-dimensional image of an organ or a tissue.

SPECT involves detection of gamma rays emitted singly, and sequentially, by the radionuclide provided by the radiolabeled chemical. To acquire SPECT images, the gamma camera is rotated around the patient. Projections are acquired at defined points during the rotation, typically every 3-6 degrees. In most cases, a full 360 degree rotation is used to obtain an optimal reconstruction. The time taken to obtain each projection is also variable, but 15-20 seconds is typical. This gives a total scan time of 15-20 minutes. Multi-headed gamma cameras are faster.

Since SPECT acquisition is very similar to planar gamma camera imaging, the same radiopharmaceuticals may be used.

Positron Emitting Tomography (PET) is a non-invasive, diagnostic imaging technique for measuring the biochemical status or metabolic activity of cells within the human body. PET is unique since it produces images of the body's basic biochemistry or functions. Traditional diagnostic techniques, such as X-rays, CT scans, or MRI, produce images of the body's anatomy or structure. The premise with these techniques is that any changes in structure or anatomy associated with a disease can be seen. Biochemical processes are also altered by a disease, and may occur before any gross changes in anatomy. PET is an imaging technique that can visualize some of these early biochemical changes. PET scanners rely on radiation emitted from the patient to create the images. Each patient is given a minute amount of a radioactive pharmaceutical that either closely resembles a natural substance used by the body or binds specifically to a receptor or molecular structure. As the radioisotope undergoes positron emission decay (also known as positive beta decay), it emits a positron, the antiparticle counterpart of an electron. After traveling up to a few millimeters, the positron encounters an electron and annihilates, producing a pair of annihilation (gamma) photons moving in opposite directions. These are detected when they reach a scintillation material in the scanning device, creating a burst of light, which is detected by photomultiplier tubes or silicon avalanche photodiodes. The technique depends on simultaneous or coincident detection of the pair of photons. Photons that do not arrive in pairs, i.e., within a few nanoseconds, are ignored. All coincidences are forwarded to the image processing unit where the final image data is produced using image reconstruction procedures.

SPECT/CT and PET/CT is the combination of SPECT and PET with computed tomography (CT). The key benefits of combining these modalities are improving the reader's confidence and accuracy. With traditional PET and SPECT, the limited number of photons emitted from the area of abnormality produces a very low-level background that makes it difficult to anatomically localize the area. Adding CT helps determine the location of the abnormal area from an anatomic perspective and categorize the likelihood that this represents a disease.

It is within the present inventions that the method for the diagnosis of a disease of the invention may realize each and any of the above strategies which are as such known in the art, and which insofar constitute further embodiments of the invention.

Compounds of the present invention are useful to stratify patients, i.e. to create subsets within a patient population that provide more detailed information about how the patient will respond to a given drug. Stratification can be a critical component to transforming a clinical trial from a negative or neutral outcome to one with a positive outcome by identifying the subset of the population most likely to respond to a novel therapy.

Stratification includes the identification of a group of patients with shared “biological” characteristics to select the optimal management for the patients and achieve the best possible outcome in terms of risk assessment, risk prevention and achievement of the optimal treatment outcome.

A compound of the present invention may be used to assess or detect, a specific disease as early as possible (which is a diagnostic use), the risk of developing a disease (which is a susceptibility/risk use), the evolution of a disease including indolent vs. aggressive (which is a prognostic use) and it may be used to predict the response and the toxicity to a given treatment (which is a predictive use).

It is also within the present invention that the compound of the invention is used in a theragnostic method. The concept of theragnostics is to combine a therapeutic agent with a corresponding diagnostic test that can increase the clinical use of the therapeutic drug. The concept of theragnostics is becoming increasingly attractive and is widely considered the key to improving the efficiency of drug treatment by helping doctors identify patients who might profit from a given therapy and hence avoid unnecessary treatments.

The concept of theragnostics is to combine a therapeutic agent with a diagnostic test that allows doctors to identify those patients who will benefit most from a given therapy. In an embodiment and as preferably used herein, a compound of the present invention is used for the diagnosis of a patient, i.e. identification and localization of the primary tumor mass as well as potential local and distant metastases. Furthermore, the tumor volume can be determined, especially utilizing three-dimensional diagnostic modalities such as SPECT or PET. Only those patients having FAP-positive tumor masses and who, therefore, might profit from a given therapy are selected for a particular therapy and hence unnecessary treatments are avoided. Preferably, such therapy is a FAP-targeted therapy using a compound of the present invention. In one particular embodiment, chemically identical tumor-targeted diagnostics, preferably imaging diagnostics for scintigraphy, PET or SPECT and radiotherapeutics are applied. Such compounds only differ in the radionuclide and therefore usually have a very similar if not identical pharmacokinetic profile. This can be realized using a chelator and a diagnostic or therapeutic radiometal.

Alternatively, this can be realized using a precursor for radiolabeling and radiolabeling with either a diagnostic or a therapeutic radionuclide. In one embodiment diagnostic imaging is used preferably by means of quantification of the radiation of the diagnostic radionuclide and subsequent dosimetry which is known to those skilled in the art and the prediction of drug concentrations in the tumor compared to vulnerable side effect organs. Thus, a truly individualized drug dosing therapy for the patient is achieved.

In an embodiment and as preferably used herein, the theragnostic method is realized with only one theragnostically active compound such as a compound of the present invention labeled with a radionuclide emitting diagnostically detectable radiation (e.g. positrons or gamma rays) as well as therapeutically effective radiation (e.g. electrons or alpha particles).

The invention also contemplates a method of intraoperatively identifying/disclosing diseased tissues expressing FAP in a subject. Such method uses a compound of the invention, whereby such compound of the invention preferably comprises as Effector a diagnostically active agent.

According to a further embodiment of the invention, the compound of the invention, particularly if complexed with a radionuclide, may be employed as adjunct or adjuvant to any other tumor treatment including, surgery as the primary method of treatment of most isolated solid cancers, radiation therapy involving the use of ionizing radiation in an attempt to either cure or improve the symptoms of cancer using either sealed internal sources in the form of brachytherapy or external sources, chemotherapy such as alkylating agents, antimetabolites, anthracyclines, plant alkaloids, topoisomerase inhibitors, and other antitumor agents, hormone treatments that modulate tumor cell behavior without directly attacking those cells, targeted agents which directly target a molecular abnormality in certain types of cancer including monoclonal antibodies and tyrosine kinase inhibitors, angiogenesis inhibitors, immunotherapy, cancer vaccination, palliative care including actions to reduce the physical, emotional, spiritual, and psycho-social distress to improve the patient's quality of life and alternative treatments including a diverse group of health care systems, practices, and products that are not part of conventional medicine.

In an embodiment of the methods of the invention, the subject is a patient. In an embodiment, a patient is a subject which has been diagnosed as suffering from or which is suspected of suffering from or which is at risk of suffering from or developing a disease, whereby the disease is a disease as described herein and preferably a disease involving FAP.

Dosages employed in practicing the methods for treatment and diagnosis, respectively, where a radionuclide is used and more specifically attached to or part of the compound of the invention will vary depending e.g. on the particular condition to be treated, for example the known radiosensitivity of the tumor type, the volume of the tumor and the therapy desired. In general, the dose is calculated on the basis of radioactivity distribution to each organ and on observed target uptake. A γ-emitting complex may be administered once or at several times for diagnostic imaging. In animals, an indicated dose range may be from 0.1 μg/kg to 5 mg/kg of the compound of the invention complexed e.g. with 1 to 200 MBq of 11In or ⁸⁹Zr. A β-emitting complex of the compound of the invention may be administered at several time points e.g. over a period of 1 to 3 weeks or longer. In animals, an indicated dosage range may be of from 0.1 μg/kg to 5 mg/kg of the compound of the invention complexed e.g. with 1 to 200 MBq ⁹⁰Y or ¹⁷⁷Lu. In larger mammals, for example humans, an indicated dosage range is from 0.1 to 100 μg/kg of the compound of the invention complexed with e.g. 10 to 400 MBq ¹¹¹In or ⁸⁹Zr. In larger mammals, for example humans, an indicated dosage range is of from 0.1 to 100 μg/kg of the compound of the invention complexed with e.g. 10 to 5000 MBq ⁹⁰Y or ¹⁷⁷Lu.

In a further aspect, the instant invention is related to a composition and a pharmaceutical composition in particular, comprising the compound of the invention.

The pharmaceutical composition of the present invention comprises at least one compound of the invention and, optionally, one or more carrier substances, excipients and/or adjuvants. The pharmaceutical composition may additionally comprise, for example, one or more of water, buffers such as, e.g., neutral buffered saline or phosphate buffered saline, ethanol, mineral oil, vegetable oil, dimethylsulfoxide, carbohydrates such as e.g., glucose, mannose, sucrose or dextrans, mannitol, proteins, adjuvants, polypeptides or amino acids such as glycine, antioxidants, chelating agents such as EDTA or glutathione and/or preservatives. Furthermore, one or more other active ingredients may, but need not, be included in the pharmaceutical composition of the invention.

The pharmaceutical composition of the invention may be formulated for any appropriate route of administration, including, for example, topical such as, e.g., transdermal or ocular, oral, buccal, nasal, vaginal, rectal or parenteral administration. The term parenteral as used herein includes subcutaneous, intradermal, intravascular such as, e.g., intravenous, intramuscular, intrathecal and intraperitoneal injection, as well as any similar injection or infusion technique. A preferred route of administration is intravenous administration.

In an embodiment of the invention the compound of the invention comprising a radionuclide is administered by any conventional route, in particular intravenously, e.g. in the form of injectable solutions or suspensions. The compound of the invention may also be administered advantageously by infusion, e.g., by an infusion of 30 to 60 min.

Depending on the site of the tumor, the compound of the invention may be administered as close as possible to the tumor site, e.g. by means of a catheter. Such administration may be carried out directly into the tumor tissue or into the surrounding tissue or into the afferent blood vessels. The compound of the invention may also be administered repeatedly in doses, preferably in divided doses.

According to a preferred embodiment of the invention, a pharmaceutical composition of the invention comprises a stabilizer, e.g. a free radical scavenger, which inhibits autoradiolysis of the compound of the invention. Suitable stabilizers include, e.g., serum albumin, ascorbic acid, retinol, gentisic acid or a derivative thereof, or an amino acid infusion solution such, e.g., used for parenteral protein feeding, preferably free from electrolyte and glucose, for example a commercially available amino acid infusion such as Proteinsteril® KE Nephro. Ascorbic acid and gentisic acid are preferred.

A pharmaceutical composition of the invention may comprise further additives, e.g. an agent to adjust the pH between 7.2 and 7.4, e.g. sodium or ammonium acetate or Na₂HPO₄. Preferably, the stabilizer is added to the non-radioactive compound of the invention and introduction of the radionuclide, for instance the complexation with the radionuclide, is performed in the presence of the stabilizer, either at room temperature or, preferably, at a temperature of from 40 to 120° C. The complexation may conveniently be performed under air free conditions, e.g. under N₂or Ar. Further stabilizer may be added to the composition after complexation.

Excretion of the compound of the invention, particularly if the Effector is a radionuclide, essentially takes place through the kidneys. Further protection of the kidneys from radioactivity accumulation may be achieved by administration of lysine or arginine or an amino acid solution having a high content of lysine and/or arginine, e.g. a commercially available amino acid solution such as Synthamin®-14 or -10, prior to the injection of or together with the compound of the invention, particularly if the Effector is a radionuclide. Protection of the kidneys may also be achieved by administration of plasma expanders such as e.g. gelofusine, either instead of or in addition to amino acid infusion. Protection of the kidneys may also be achieved by administration of diuretics providing a means of forced diuresis which elevates the rate of urination. Such diuretics include high ceiling loop diuretics, thiazides, carbonic anhydrase inhibitors, potassium-sparing diuretics, calcium-sparing diuretics, osmotic diuretics and low ceiling diuretics. A pharmaceutical composition of the invention may contain, apart from a compound of the invention, at least one of these further compounds intended for or suitable for kidney protection, preferably kidney protection of the subject to which the compound of the invention is administered.

It will be understood by a person skilled in the art that the compound of the invention is disclosed herein for use in various methods. It will be further understood by a person skilled in the art that the composition of the invention and the pharmaceutical composition of the invention can be equally used in said various methods. It will also be understood by a person skilled in the art that the composition of the invention and the pharmaceutical composition are disclosed herein for use in various methods. It will be equally understood by a person skilled in the art that the compound of the invention can be equally used in said various methods.

It will be acknowledged by a person skilled in the art that the composition of the invention and the pharmaceutical composition of the invention contain one or more further compounds in addition to the compound of the invention. To the extent that such one or more further compounds are disclosed herein as being part of the composition of the invention and/or of the pharmaceutical composition of the invention, it will be understood that such one or more further compounds can be administered separately from the compound of the invention to the subject which is exposed to or the subject of a method of the invention. Such administration of the one or more further compounds can be performed prior, concurrently with or after the administration of the compound of the invention. It will also be acknowledged by a person skilled in the art that in a method of the invention, apart from a compound of the invention, one or more further compound may be administered to a subject. Such administration of the one or more further compounds can be performed prior, concurrently with or after the administration of the compound of the invention. To the extent that such one or more further compounds are disclosed herein as being administered as part of a method of the invention, it will be understood that such one or more further compounds are part of a composition of the invention and/or of a pharmaceutical composition of the invention. It is within the present invention that the compound of the invention and the one or more further compounds may be contained in the same or a different formulation. It is also within the present invention that the compound of the invention and the one or more further compounds are not contained in the same formulation, but are contained in the same package containing a first formulation comprising a compound of the invention, and a second formulation comprising the one or more further compounds, whereby the type of formulation may be the same or may be different.

It is within the present invention that more than one type of a compound of the invention is contained in the composition of the invention and/or the pharmaceutical composition of the invention. It is also within the present invention that more than one type of a compound of the invention is used, preferably administered, in a method of the invention.

It will be acknowledged that a composition of the invention and a pharmaceutical composition of the invention may be manufactured in conventional manner.

Radiopharmaceuticals have decreasing content of radioactivity with time, as a consequence of the radioactive decay. The physical half-life of the radionuclide is often short for radiopharmaceutical diagnostics. In these cases, the final preparation has to be done shortly before administration to the patient. This is in particular the case for positron emitting radiopharmaceuticals for tomography (PET radiopharmaceuticals). It often leads to the use of semi-manufactured products such as radionuclide generators, radioactive precursors and kits.

Preferably, a kit of the invention comprises apart from one or more than one compounds of the invention typically at least one of the followings: instructions for use, final preparation and/or quality control, one or more optional excipient(s), one or more optional reagents for the labeling procedure, optionally one or more radionuclide(s) with or without shielded containers, and optionally one or more device(s), whereby the device(s) is/are selected from the group comprising a labeling device, a purification device, an analytical device, a handling device, a radioprotection device or an administration device.

Shielded containers known as “pigs” for general handling and transport of radiopharmaceutical containers come in various configurations for holding radiopharmaceutical containers such as bottles, vials, syringes, etc. One form often includes a removable cover that allows access to the held radiopharmaceutical container. When the pig cover is in place, the radiation exposure is acceptable.

A labeling device is selected from the group of open reactors, closed reactors, microfluidic systems, nanoreactors, cartridges, pressure vessels, vials, temperature controllable reactors, mixing or shaking reactors and combinations thereof.

A purification device is preferably selected from the group of ion exchange chromatography columns or devices, size-exclusion chromatography columns or devices, affinity chromatography columns or devices, gas or liquid chromatography columns or devices, solid phase extraction columns or devices, filtering devices, centrifugations vials columns or devices.

An analytical device is preferably selected from the group of tests or test devices to determine the identity, radiochemical purity, radionuclidic purity, content of radioactivity and specific radioactivity of the radiolabelled compound.

A handling device is preferably selected from the group consisting of devices for mixing, diluting, dispensing, labeling, injecting and administering radiopharmaceuticals to a subject.

A radioprotection device is used in order to protect doctors and other personnel from radiation when using therapeutic or diagnostic radionuclides. The radioprotection device is preferably selected from the group consisting of devices with protective barriers of radiation-absorbing material selected from the group consisting of aluminum, plastics, wood, lead, iron, lead glass, water, rubber, plastic, cloth, devices ensuring adequate distances from the radiation sources, devices reducing exposure time to the radionuclide, devices restricting inhalation, ingestion, or other modes of entry of radioactive material into the body and devices providing combinations of these measures.

An administration device is preferably selected from the group of syringes, shielded syringes, needles, pumps, and infusion devices. Syringe shields are commonly hollow cylindrical structures that accommodate the cylindrical body of the syringe and are constructed of lead or tungsten with a lead glass window that allows the handler to view the syringe plunger and liquid volume within the syringe.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention is now further illustrated by reference to the following figures and examples from which further features, embodiments and advantages, may be taken, wherein

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;

FIG. 2 shows a radiochromatogram of ¹⁷⁷Lu-3BP-3407 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed six days after synthesis;

FIG. 3 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;

FIG. 4 shows a radiochromatogram of ¹⁷⁷Lu-3BP-3554 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed six days after synthesis;

FIG. 5A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3105 1 h 3h 6 h and 24h post injection into the mouse model;

FIG. 5B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3168 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 6A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3320 1 h 3h, 6 h and 24h post injection into the mouse model;

FIG. 6B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3321 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 7A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3275 1 h 3h, 6 h and 24h post injection into the mouse model; FIG. 7B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3397-(B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 8A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3398 1 h 3h 6 h and 24h post injection into the mouse model;

FIG. 8B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3407 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 9A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3554 1 h 3h, 6 h and 24h post injection into the mouse model;

FIG. 9B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3652 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 10A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3654 1 h 3h, 6 h and 24h post injection into the mouse model;

FIG. 10B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3656 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 11A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3659 h 3h 6 h and 24h post injection into the mouse model;

FIG. 11B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3678 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 12A shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3692 h 3h 6 h and 24h post injection into the mouse model;

FIG. 12B shows the percentage of injected dose per gram of tissue (% ID/g) uptake in the kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3767 (B) 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 13 shows SPECT-images of ¹¹¹In-3BP-3554 1 h, 3 h, 6 h, 24 h and 48 h post injection into mice with HEK-FAP tumors

FIG. 14 shows SPECT-images of ¹¹¹In-3BP-3767 1 h, 3 h, 6 h, 24 h and 48 h post injection into mice with HEK-FAP tumors;

FIG. 15-A shows tumor growth over time in mice with HEKFAP tumprs treated with vehicle, cold compound ^natLu-3BP-3554, 30 MBq (low dose)¹⁷⁷Lu-3BP-3554, and 60 MBq (high dose) ¹⁷⁷Lu-3BP-3554;

FIG. 15-B shows percent body weight changes over time in mice with HEK-FAP tumors treated with vehicle, cold compound ^natLu-3BP-3554, 30 MBq (low dose)¹⁷⁷Lu-3BP-3554, and 60 MBq (high dose)¹⁷⁷Lu-3BP-3554;

FIG. 16-A shows representative SPECT/CT images over time of the biodistribution 60 MBq ¹⁷⁷Lu-3BP-3554 in mice with HEK-FAP tumors;

FIG. 16-B shows representative SPECT/CT images over time of the biodistribution 30 MBq ¹⁷⁷Lu-3BP-3554 in mice with HEK-FAP tumors,

FIG. 17-A shows representative SPECT/CT images of four different sarcoma PDX models 3 h after ¹¹¹In-3BP-3554 administration;

FIG. 17-B shows % ID/g uptake of ¹¹¹In-3BP-3554 in four different sarcoma PDX models, 3 hours post injection;

FIG. 18-A shows tumor growth over time in mice with Sarc4809 PDX tumors treated with vehicle, cold compound ^natLu-3BP-3554, 30 MBq ¹⁷⁷Lu-3BP-3554 or 60 MBq ¹⁷⁷Lu-3BP-3554;

FIG. 18-B shows body weight changes over time in mice with sarcoma Sarc4809 PDX tumors treated with vehicle, cold compound ^natLu-3BP-3554, 30 MBq ¹⁷⁷Lu-3BP-3554, or 60 MBq ¹⁷⁷Lu-3BP-3554;

FIG. 19 shows the amino acid sequences of human fibroblast activating protein (FAP) (SEQ ID NO: 1), human dipeptidyl peptidase 4 (DDP4) (SEQ ID NO: 2) and human prolyl endopeptidase (PREP) (SEQ ID NO: 3);

FIG. 20 shows the percentage of injected dose per gram of tissue (% ID/g) uptake in kidney, liver, bloodpool and HEK-FAP tumor as determined by SPECT-imaging of ¹¹¹In-3BP-3940 1 h, 3 h, 6 h and 24h post injection into the mouse model;

FIG. 21 shows SPECT-images of ¹¹¹In-3BP-3940 1 h, 3 h, 6 h, 24 h and 48 h post injection into mice with HEK-FAP tumors;

FIG. 22 shows representative SPECT/CT-images of the biodistribution of ^99mTc-3BP-4219 at 1 h, 3 h, and 6 h post injection in mice with HEK-FAP tumors;

FIG. 23 shows representative SPECT/CT-images of the biodistribution of ^99mTc-3BP-4221 at 1 h, 3 h, and 6 h post injection in mice with HEK-FAP tumors;

FIG. 24 shows representative SPECT/CT-images of the biodistribution of ^99mTc-3BP-4541 at 1 h, 3 h, and 6 h post injection in mice with HEK-FAP tumors;

FIG. 25 shows representative SPECT/CT-images of the biodistribution of ^99mTc-3BP-4961 at 1 h, 3 h, and 6 h post injection in mice with HEK-FAP tumors;

FIG. 26 shows representative PET/CT-images of the biodistribution of ⁶⁸Ga-3BP-4768 at 0.25 h, 1 h, and 3 h post injection in mice with HEK-FAP tumors;

FIG. 27 shows representative PET/CT-images of the biodistribution of ⁶⁸Ga-3BP-5201 at 0.25 h, 1 h, and 3 h post injection in mice with HEK-FAP tumors; and

FIG. 28 shows representative SPECT/CT-images of the biodistribution of ¹¹¹In-3BP-4560 at 1 h, 3 h, and 6 h, 24 h, and 48 h post injection in mice with HEK-FAP tumors.

The following Examples have been included to provide guidance to one of ordinary skill in the art for practicing representative embodiments of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill can appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter. The synthetic descriptions and specific examples that follow are only intended for the purposes of illustration, and are not to be construed as limiting in any manner to make compounds of the disclosure by other methods.

EXAMPLES

Abbreviations used in the instant application and the following examples in particular are as follows:

- 4PL means four parameter logistic curve fitting
- A means angstrom
- ACN means acetonitrile
- Ahx means 6-Aminohexanoic acid
- AMC means 7-amino-4-methylcoumarin
- amu means atomic mass unit
- aq. means aqueous
- AUC_infmeans area under the curve extrapolated to infinity
- BPS means blood pool surrogate
- BSA means bovine serum albumin
- C₀means initial concentration of the compound
- CAF means cancer associated fibroblasts
- CL means clearance
- CM means ChemMatrix™
- CT means computed tomography
- Cy5 means Cyanine-5
- DAD means Diode Array Detector
- DCM means dichloromethane
- Dde means N-(1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl)
- DEG means di ethylene glycol dimethacrylate
- DIC means N,N′-Diisopropylcarbodiimide
- DICOM means Digital Imaging and Communications in Medicine
- DIPEA means diisopropylethylamine
- DMF means N,N-dimethylformamide
- DMSO means dimethyl sulfoxide
- 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
- DPP means dipeptidyl peptidase
- EC means electron capture
- EC₅₀means half-maximal excitatory concentration
- ECACC means European Collection of Authenticated Cell Cultures
- EDC means 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
- EMEM means Eagle's Minimum Essential Medium
- eq or eq. means equivalent
- ESI means electrospray ionization
- Et₂O means Diethylether
- EtOAc means ethylacetate
- FACS means fluorescence-activated cell sorting
- FAP means fibroblast activation protein
- Fb means background fluorescent intensity
- FBS means fetal bovine serum
- FGF21 means fibroblast growth factor 21
- FITC means 5(6)-fluorescein isothiocyanate
- Fmoc means 9-Fluorenylmethoxycarbonyl
- FRET means Fluorescence Resonance Energy Transfer
- Ft means fluorescent intensity
- Gab means gamma-amino butyric acid
- GABA means gamma-amino butyric acid
- h means hour(s)
- HATU means O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate
- HBST means SPR running buffer
- HEK-FAP means human embryonic kidney 293 cells expressing human FAP
- HEPES means 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- HFIP means hexafluoro-2-isopanol
- HOAc means acetic acid
- HOAt means 1-Hydroxy-7-azabenzotriazole
- HPLC means high performance liquid chromatography
- HPLC/MS means high performance liquid chromatography/mass spectrometry
- IC₅₀means half-maximal inhibitory concentration
- ID/g means injected dose per gram
- IS means isomeric transition
- iTLC-SG means instant thin layer chromatography-silica-gel
- K2EDTA means ethylenediaminetetraacetic acid dipotassium
- K_Dmeans dissociation constant
- kDa means 1000 Dalton
- K_imeans inhibitory constant
- k_offmeans dissociation rate
- k_onmeans association rate
- LC/TOF-MS means Liquid chromatography/time-of-flight/mass spectrometry
- LC-MS means high performance liquid chromatography coupled with mass spectrometry
- LDH means lactate dehydrogenase
- Leu means leucine
- LiOH means lithium hydroxide
- M means molar or mol per Liter
- m/z means mass divided by charge
- max. means maximum
- MeOH means Methanol
- MeV means mega electron volt
- min means minute(s)
- MMP means matrix metalloproteinase
- MRM means multiple reaction monitoring
- MTBE means Methyl-tert-butylether
- Mtt means Methyltrityl
- MTV means mean tumor volume
- MW means molecular weight
- n.d. means not determined
- Na₂SO₄means sodium sulfate
- NaCl means sodium chloride
- NaHCO₃means sodium hydrogencarbonate
- NCA means non-compartmental analysis
- NHS means N-Hydroxysuccinimide
- NMP means 1-methyl-2-pyrrolidone
- NOS means not otherwise specified
- Oic means L-octahydroindol-2-carbonsaure
- p.a. means: for analytical purpose (quality grade)
- p.i. means post injection
- Pbf means 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-sulfonyl
- PBS means phosphate buffered saline
- PDX means patient-derived xenograft
- PET means positron emission tomography
- pIC50 means the negative log of the IC50 value when converted to molar
- POP means prolyl oligopeptidase
- ppm means parts per million
- PREP means prolyl endopeptidase
- prep. means preparative
- PS means polystyrene
- Q-TOF means quadrupole time of flight
- Ref means reference
- RFU means relative fluorescence unit
- RLB means radioligand binding assay
- RMCE means recombinase-mediated cassette exchange
- RP means reversed phase
- R_tmeans retention time
- RT means room temperature
- RU means resonance units
- SAR means structure activity relationship
- sat. means saturated
- SCID means severe combined immunodeficiency
- SCK means single cycle kinetics
- sec or s means second
- SF means spontaneous fission
- SPECT means single photon emission computed tomography
- SPPS means Solid Phase Peptide Synthesis
- t_1/2means terminal half-life
- tBu means tert. butyl
- TFA means trifluoroacetate or trifluoroacetic acid
- TG means TentaGel
- TGI means tumor growth inhibition
- THF means Tetrahydrofuran
- TIPS means triisopropylsilane
- TLC means thin layer chromatography
- TME means tumor microenvironment
- t_Rmeans retention time
- UHPLC means ultrahigh performance liquid chromatography
- UV means ultraviolet
- V_SSmeans volume of distribution at steady state
- V_Zmeans volume of distribution in the terminal phase

Example 1: Material and Methods

The materials and methods as well as general methods are further illustrated by the following examples.

Solvents:

Solvents were used in the specified quality without further purification. Acetonitrile (Super Gradient, HPLC, VWR—for analytical purposes; PrepSolv, Merck—for preparative purposes); dichloromethane (synthesis, Roth); ethyl acetate (synthesis grade, Roth); N,N-dimethylformamide (peptide synthesis grade, Biosolve); 1-methyl-2-pyrolidone (peptide grade, IRIS BioTech) 1,4-dioxane (reinst, Roth); methanol (p. a., Merck).

Water: Milli-Q Plus, Millipore, demineralized.

Chemicals:

Chemicals were synthesized according to or in analogy to literature procedures or purchased from Sigma-Aldrich-Merck (Deisenhofen, Germany), Bachem (Bubendorf, Switzerland), VWR (Darmstadt, Germany), Novabiochem (Merck Group, Darmstadt, Germany), Acros Organics (distribution company Fisher Scientific GmbH, Schwerte, Germany), Iris Biotech (Marktredwitz, Germany), Amatek Chemical (Jiangsu, China), Roth (Karlsruhe, Deutschland), Molecular Devices (Chicago, USA), Biochrom (Berlin, Germany), Peptech (Cambridge, Mass., 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, Calif., USA) or other companies and used in the assigned quality without further purification.

Boc₄N4Ac—OH was synthesized according to a literature procedure (Maecke et al. Chem. Eur. J., 2010, 16, 7, 2115).

embedded image

Cells:

HT29 (ECACC Cat. No. 91072201) and WI-38 (ECACC Cat. No. 90020107) were purchased from ECACC and HEK293 cells expressing human FAP (Q12884) were produced by InSCREENeX GmbH (Braunschweig, Germany) using recombinase-mediated cassette exchange (RMCE). The RMCE procedure is described by Nehlsen et al. (Nehlsen, et al., BMC Biotechnol, 2009, 9: 100).

HPLC/MS Analyses

HPLC/MS analyses were performed by injection of 5 μl of a solution of the sample, using a 2 step gradient for all chromatograms (5-65% B in 12 min, followed by 65-90% in 0.5 min, A: 0.1% TFA in water and B: 0.1% TFA in ACN). RP columns were from Agilent (Type Poroshell 120, 2.7 μm, EC-C18, 50×3.00 mm, flow 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 done at λ=230 nm. Retention times (R_t) are indicated in the decimal system (e.g. 1.9 min=1 min 54 s) and are referring to detection in the UV spectrometer. For the evaluation of observed compound masses the ‘Find Compounds by Formula’-feature was used. In particular, the individual ‘neutral mass of a compound (in units of Daltons)’-values and the corresponding isotope distribution pattern were used to confirm compound identity. The accuracy of the mass spectrometer was approx. ±5 ppm.

Preparative HPLC:

Preparative HPLC separations were done with reversed phase columns (Kinetex 5μ XB-C18 100 Å, 150×30 mm from Phenomenex or RLRP-S 8μ, 100 Å, 150×25 mm) as stationary phase. As mobile phase 0.1% TFA in water (A) and 0.1% TFA in ACN (B) were used which were mixed in linear binary gradients. The gradients are described as: “10 to 40% B in 30 min”, which means a linear gradient from 10% B (and correspondingly 90% A) to 40% B (and correspondingly 60% A) was run within 30 min. Flow-rates were within the range of 30 to 50 ml/min. A typical gradient for the purification of the compounds of the invention started at 5-25% B and ended after 30 min at 35-50% B and the difference between the percentage B at end and start was at least 10%. A commonly used gradient was “15 to 40% B in 30 min”.

General Procedures for Automated/Semi-Automated Solid-Phase Synthesis:

Automated solid-phase of peptides and polyamides was performed on a Tetras Peptide Synthesizer (Advanced ChemTech) in 50 μmol and 100 μmol scales. Manual steps were performed in plastic syringes equipped with frits (material PE, Roland Vetter Laborbedarf OHG, Ammerbuch, Germany). The amount of reagents in the protocols described corresponds to the 100 μmol scale, unless stated otherwise.

Solid-phase synthesis was performed on polystyrene (cross linked 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.

Resin Loading:

In case of the trityl linker the 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 minutes and subsequently washed with DCM (3 ml, 1 minute). Then the resin was 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 hour. Afterwards the resin was washed with methanol (5 ml, 5 minutes) and DMF (3 ml, 2×1 minute).

In case of the Wang linker pre-loaded resins (polystyrene (PS) and TentaGel (TG)) were employed.

In case of the rink amide linker the attachment of the first residue the resin (CM, DEG) was performed with the same procedure as for the chain assembly as described below.

Alloc/Allyl-Deprotection:

After swelling in DMF, the resin was washed with DMF and DCM. DCM was de-oxygenated by passing a stream of nitrogen through the stirred solvent. The oxygen-free solvent was used to wash the resin trice. Then 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 added to the resin. The resin was agitated 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 3 times with 3 ml, 1 minute).

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 minutes) and subsequently washed with DMF (3 ml, 5×1 minute).

Dde-Deprotection:

After swelling in DMF, the resin was washed with DMF and then treated with hydrazine-hydrate/DMF (2/98, 3 ml 2×10 minutes) and subsequently washed with DMF (3 ml, 5×1 minute).

Mtt-Deprotection:

After swelling in DCM, the resin was washed with DCM and then treated with HFIP/DCM (7/3, 4-6 ml, 4 hours) and subsequently washed with DCM (3 ml, 3×1 minute), DMF (3 ml, 3×1 ml) and DIPEA (0.9 M in DMF, 3 ml, 1 minute).

Solutions of Reagents:

Building Blocks (0.3 M in DMF or NMP), DIPEA (0.9 M in DMF), HATU (0.4 M in DMF), Acetic anhydride (0.75 M in DMF)

Coupling: Coupling of Building Blocks/Amino Acids (Chain Assembly):

Unless otherwise stated, coupling of building blocks was performed as follows: After subsequent addition of solutions of the corresponding building block (1.7 ml, 5eq.), DIPEA solution (1.15 ml, 10 eq.) and 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 minute) and the coupling step was repeated.

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 minutes. Afterwards the resin was washed with DMF (3 ml, 6×1 minutes).

Cleavage Method A: Cleavage of Protected Fragments from Hyper-Acid Labile Resin:

After the completion of the assembly of the sequence the resin was finally washed with DCM (3 ml, 4×1 minute) and then dried in the vacuum. Then the resin was treated with HFIP/DCM (7/1, 4 ml, 4 hours) and the collected solution evaporated to dryness. The residue was purified with preparative HPLC or used without further purification.

Cleavage Method B: Cleavage of Unprotected Fragments (Complete Resin Cleavage):

After the completion of the assembly of the sequence the resin was finally washed with DCM (3 ml, 4×1 minute), dried in the vacuum overnight and treated with TFA, EDT, water and TIPS (94/2.5/2.5/1) for 2 h (unless otherwise stated). Afterwards the cleavage solution was poured into a chilled mixture of MTBE and cyclohexane (1/1, 10-fold excess compared to the volume of cleavage solution), centrifuged at 4° C. for 5 minutes and the precipitate collected and dried in the vacuum. The residue was lyophilized from water/acetonitrile prior to purification or further modification.

Cleavage Method C: Cleavage of Protective Groups of Peptides in Solution

The protected/partially protected compound was dissolved in TFA, water and TIPS (95/2.5/2.5) for 2 h (unless otherwise stated). Afterwards the cleavage solution was poured into a chilled mixture of MTBE and cyclohexane (1/1, 10-fold excess compared to the volume of cleavage solution), centrifuged at 4° C. for 5 minutes and the precipitate collected and dried in the vacuum. The residue was lyophilized from water/acetonitrile prior to purification or further modification.

More relevant Fmoc-solid-phase-peptide synthesis methods are 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 plugin (Mestrelab Research, S.L.), or AutoNom version 2.2 (Beilstein Informationssysteme Copyright© 1988-1998, Beilstein Institut für Literatur der Organischen Chemie licensed to Beilstein Chemiedaten and Software GmbH, where appropriate.

Preparation of Compounds:

Specific embodiments for the preparation of compounds of the invention are provided in the following examples. Unless otherwise specified, all starting materials and reagents are of standard commercial grade, and are used without further purification, or are readily prepared from such materials by routine methods. Those skilled in the art of organic synthesis will recognize in light of the instant disclosure that starting materials and reaction conditions may be varied including additional steps employed to produce compounds encompassed by the present invention.

One general synthesis route for compounds of the invention comprises

- 1. Solid Phase Peptide Synthesis (SPPS) of a linear peptide precursor with two thiol moieties.
- 2. the thiol-site specific cyclization of this linear peptide precursor with
  - a. a bis(bromomethyl)benzene derivative or
  - b. a tris(bromomethyl)benzene derivative.
- 3. In case of cyclizations with a tris(bromomethyl)benzene derivative the intermediate formed in the cyclization reaction was further reacted with a linker that enabled the attachment of a chelator.

Example 2: Synthesis of Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-His-Nmf-Arg-Asp-NH₂(3BP-3188)

The sequence (Ac-Met-Cys-Pro-Pro-Thr-Glu-Phe-Cys-Asp-His-Nmf-Arg-Asp-NH₂) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Rink amide resin. After performing the steps of ‘Cleavage method B’ the lyophilized crude peptide residue was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH=8.5) and acetonitrile. To this mixture a solution of 14.5 mg α,α′-dibromo-m-xylene (55 μmol, 1.1 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. Upon completion of the cyclization reaction 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 8.61 mg of the pure title compound (9.8%). HPLC: R_t=5.87 min. LC/TOF-MS: exact mass 1753.716 (calculated 1753.705). C₇₉H₁₀₇N₁₉O₁₁S₃(MW=1755.011).

Example 3: Synthesis of DOTA-Ttds-Leu-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-His-Phe-Arg-Asp-NH₂(3BP-3172)

The sequence (DOTA-Ttds-Leu-Cys-Pro-Pro-Thr-Glu-Phe-Cys-Asp-His-Phe-Arg-Asp-NH₂) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Rink amide resin. After performing the steps of ‘Cleavage method B’ the lyophilized crude peptide residue was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH=8.5) and acetonitrile. To this mixture a solution of 14.5 mg α,α′-dibromo-m-xylene (55 μmol, 1.1 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. Upon completion of the cyclization reaction 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (20 to 45% B in 30 min—Kinetex) to yield 35.46 mg of the pure title compound (29.8%). HPLC: R_t=5.9 min. LC/TOF-MS: exact mass 2368.091 (calculated 2368.087). C₁₀₇H₁₅₇N₂₅O₃₂S₂(MW=2369.676).

Example 4: Synthesis of Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-His-Ppa-arg-Ttds-Lys(DOTA)-NH₂(3BP-3277)

The sequence (Hex-Cys-Pro-Pro-Thr-Glu-Phe-Cys-Asp-His-Ppa-arg-Ttds-Lys(Mtt)-NH₂) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Rink amide resin. Then a ‘Mtt deprotection’ described in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ was performed to liberate the s-amino function of the C-terminal lysine residue of the peptide resin. DOTA(tBu)₃-OH (143.3 mg, 250 μmol, 5 eq compared to the initial resin loading) was dissolved in 0.6 ml of a 0.4 M solution of HATU in DMF and 0.65 ml of a 0.9 M of DIPEA in DMF. After leaving the mixture for 1 minute for pre-activation it was added to the resin. An hour later 0.2 ml of a 3.2 M of DIC in DMF was added and the gentle agitation of the resin continued for a further hour. The resin was thoroughly washed and subjected to the ‘Cleavage method B’ protocol. The lyophilized remainder (linear, branched peptide Hex-Cys-Pro-Pro-Thr-Glu-Phe-Cys-Asp-His-Ppa-arg-Ttds-Lys(DOTA)-NH₂) was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH=8.5) and acetonitrile. To this mixture a solution of 14.5 mg α,α′-dibromo-m-xylene (55 μmol, 1.1 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. Upon completion of the cyclization reaction 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 17.18 mg of the pure title compound (14.5%). HPLC: R_t=5.8 min. LC/TOF-MS: exact mass 2367.150 (calculated 2367.139). C₁₀₈H₁₆₂N₂₆O₃OS₂(MW=2368.735).

Example 5: Synthesis of N4Ac-Glu(AGLU)-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4246)

The sequence (N4Ac-Glu(OAll)-Ttds-Nle-Cys-Pro-Pro-Thr-Gln-Phe-Cys-OH) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Fmoc-Cys(Trt) WANG Tentagel resin. An ‘Alloc/Allyl-deprotection’ was performed to effect the removal of the Allyl ester protecting group. 3,4;5,6-di-O-isopropylidene-1-amino-1-deoxy-D-glucitol (J. Org. Chem., 2002, 75, 3685) (52.2 mg, 200 μmol, 4 eq.), Oxyma (28.4 mg, 200 μmol, 4 eq.) and DIC (31 μL, 200 μmol, 4 eq.) were dissolved in DMF (1.5 mL), the solution added to resin and the latter agitated for 90 minutes. The resin was washed and the coupling of the amino-glucitol building block repeated one more time. The resin was washed, dried and finally treated with TFA, water, TIPS and 1,3-Dimethoxybenzol (90/2.5/2.5/5, 3 mL) for 2 hours to effect detachment from the resin and removal of the side chain protecting groups. After precipitation and lyophilization from water/acetonitrile the crude linear peptide was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH=8.5) and acetonitrile. To this mixture a solution of 14.5 mg α,α′-dibromo-m-xylene (55 μmol, 1.1 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. Upon completion of the cyclization reaction 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 8.97 mg of the pure title compound (10%). HPLC: R_t=5.5 min. LC/TOF-MS: exact mass 1789.901 (calculated 1789.899). C₈₁H₁₃₁N₁₇O₂₄S₂(MW=1791.142).

Example 6: Synthesis of Pentyl-SO2-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3692)

The sequence (H-Cys-Pro-Pro-Thr-Gln-Phe-Cys-Asp-NH₂) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Rink amide resin. The N-terminal sulfonamide was attached by treatment of the resin bound peptide with a solution of n-pentyl sulfonyl chloride (42.7 μl, 300 μmol, 6 eq) and 2,4,6-collidine (29.7 μl, 225 μmol, 4.5 eq). After overnight agitation the resin was thoroughly washed and subjected to the ‘Cleavage method B’ protocol. The lyophilized remainder (linear peptide Pentyl-SO2-Cys-Pro-Pro-Thr-Gln-Phe-Cys-Asp-NH₂) was dissolved in 60 ml of a 1:1 mixture of ammonium bicarbonate solution (50 mM, pH=8.5) and acetonitrile. To this mixture a solution of 26.8 mg 1,3,5-tris(bromomethyl)benzene (75 μmol, 1.5 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. After stirring the solution for 1 hour 43 mg piperazine (500 μmol, 10 eq compared to initial resin loading) were added. After 2 hours 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 9.15 mg (7.4 μmol) of the peptide intermediate Pentyl-SO2-[Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(14.7%). To the solution of the latter in 150 μl DMSO 2.5 μl DIPEA were added to adjust the pH value to approximately 7.5-8. Then 8.4 mg of DOTA-NHS (11 μmol, 1.5 eq compared to the peptide intermediate) in 100 μl DMSO were added. During the course of the LC/TOF-MS monitored reaction 2.5 μl DIPEA was added 3 times to re-adjust the pH value to the starting value. After reaction completion the solution was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 7.09 mg of the pure title compound (8.7% overall yield). HPLC: R_t=6.0 min. LC/TOF-MS: exact mass 1628.706 (calculated 1628.704). C₇₂H₁₀₈N₁₆O₂₁S₃(MW=1629.924).

Example 7: Synthesis of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4089)
Example 7a: Synthesis of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4089) by Two Different Methods

The synthesis of the title compound was either performed by initially synthesizing the linear peptide precursor on solid phase with a subsequent solution phase cyclization (either in non-aqueous solution (Method A) or in aqueous solution (Method B) or by performing all steps on solid phase. The latter approach (Example 7b) served as starting point for further derivatization.

For the first approach (Example 7a) Fmoc-Cys(Trt)-OH was loaded onto the trityl resin as described in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale. Onto this resin the sequence (Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-OH) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’. After performing the steps of ‘Cleavage method B’ the crude peptide was lyophilized and cyclized in solution by two alternative methods.

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 first 35 μl DIPEA and then 23.7 mg of 1,3,5-tris(bromomethyl)benzene (66.6 μmol, 1.3 eq compared to initial resin loading) were added. The solution was stirred for 1 hour and then 42.8 mg cysteamine (555 μmol, 11 eq compared to initial resin loading) was added. After 1 hour the solvents was removed in vacuo and 25 ml of a 1:1 mixture of acetonitrile and water (containing 50 μl TFA were added). The solvents were removed by lyophilization. The remainder was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 17.8 mg (16.4 μmol) of the intermediate Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (32.8%).

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 a solution of 26.8 mg 1,3,5-tris(bromomethyl)benzene (75 μmol, 1.5 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. The solution was stirred for 1 hour and then 38.6 mg cysteamine (500 μmol, 10 eq compared to initial resin loading) was added. After 2 hours 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 19.47 mg (18 μmol) of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (35.9%).

Both solution-based cyclization methods perform similar and achieve comparable yields and similar purities.

Example 7b: Synthesis of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel (3BP-4089 Bound on Peptide Resin)

For the synthesis of the resin bound title compound a Fmoc-Cys(Trt)-WANG Tentagel resin was used as starting material. Onto the latter the sequence (Hex-Cys(Trt)-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys-OH) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 1 mmol scale. After completion of the sequence assembly the resin was washed with DCM (3×1 min) Then the trityl protecting groups were selectively removed from the resin be treatment with a solution of TFA, TIPS and DCM (5/5/90, 5×5 min). The resin was washed with DCM, DMF, 0.9 M DIPEA in DMF, DMF, DCM (3/3/2/3/3) and dried in the vacuum. The following cyclization was performed in 200 μmol portions. To this end 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 DMF at 50° C. for 90 minutes. The solution was removed, the resin washed with DMF and then a solution of cysteamine (154.3 mg, 2 mmol, 10 eq) added to the resin. The resin was agitated for another 90 minutes 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 and kept for further derivatization. By this procedure it may happen that the Trityl-group at Glutamine is either partially or fully deprotected. In any case this does not interfere with the optional derivatization of the free amino group of AET.

Example 8a): Synthesis of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3554)

To the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (19.5 mg, 18 μmol, 3BP-4089—described in Example 7a) in 300 μl DMSO, 5 μl DIPEA were added to adjust the pH value to approximately 7.5-8. Then 20.5 mg of DOTA-NHS (27 μmol, 1.5 eq compared to the peptide intermediate) in 200 μl DMSO were added. During the course of the LC/TOF-MS monitored reaction 5 μl DIPEA was added 3 times to re-adjust the pH value to the starting value. After reaction completion the solution was subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 20.44 mg of the pure title compound (77.4% yield). HPLC: R_t=5.9 min. LC/TOF-MS: exact mass 1469.640 (calculated 1469.639). C₆₇H₉₉N₁₃O₁₈S₃(MW=1470.780).

Example 8b): Synthesis of nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3940)

Similar methods to the synthesis of 3BP-3554 (Example 7a, Cyclization method A and Example 8a)) were used for the synthesis of the title compound. The only difference was that after assembling of the linear peptide sequence a terminal urea moiety was introduced by the overnight reaction of butyl isocyanate (5 eq) and DIPEA (10 eq) in DMF at room temperature. The cyclization step and DOTA introduction was performed by identical methods.

After HPLC purification (15 to 40% B in 30 min—Kinetex) 28.28 mg of the pure title compound (25.6% yield) were isolated. HPLC: R_t=5.8 min. LC/TOF-MS: exact mass 1470.644 (calculated 1470.635). C₆₆H₉₈N₁₄O₁₈S₃(MW=1471.768).

Example 9: Synthesis of Hex-[Cys-(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4162)

(R)-NODA-GA(tBu)₃-OH (50 mg, 92 μmol, 1 eq), HATU (35 mg, 92 μmol, 1 eq) and DIPEA (32 μL, 184 μmol, 2 eq) were dissolved in 0.4 mL DMF. The mixture was stirred for 2 min to ensure pre-activation of the chelator building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (100 mg, 92 μmol, 3BP-4089—described in Example 7a) in 2 mL DMF and 20 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 90 min all volatiles were removed in the vacuum and the remainder subjected to lyophilization. After performing the steps of ‘Cleavage method C’ the crude peptide was lyophilized and subsequently subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 48.54 mg of the pure title compound (33.7% yield). HPLC: R_t=6.8 min. LC/TOF-MS: exact mass 1440.613 (calculated 1440.613). C₆₆H₉₆N₁₂O₁₈S₃(MW=1441.739).

Example 10: Synthesis of Hex-[Cys-(tMeBn(DTPA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4214)

DTPA(tBu)₄-OH (Diethylenetriamine-N,N,N″,N″-tetra-tert-butyl acetate-N′-acetic acid) (28.5 mg, 46 μmol, 1 eq), HATU (17.5 mg, 46 μmol, 1 eq) and DIPEA (16 μL, 92 μmol, 2 eq) were dissolved in 100 μL DMF. The mixture was stirred for 2 min to ensure pre-activation of the chelator building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (50 mg, 46 μmol, 3BP-4089—described in Example 7a) in 600 μL DMF and 10 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 180 min all volatiles were removed in the vacuum and the remainder subjected to lyophilization. After performing the steps of ‘Cleavage method C’ the crude peptide was lyophilized and subsequently subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 15.4 mg of the pure title compound (22.9% yield). HPLC: R_t=6.5 min. LC/TOF-MS: exact mass 1458.587 (calculated 1458.587). C₆₅H₉₄N₁₂O₂OS₃(MW=1459.711).

Example 11: Synthesis of Hex-[Cys-(tMeBn(N4Ac-02Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4088)

Fmoc-O2Oc-OH was loaded onto the trityl resin as described in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 100 μmol scale. Onto this resin the sequence Boc₄N4Ac—OH was coupled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’. After performing the steps of ‘Cleavage method A’ the crude protected conjugated was lyophilized (crude yield 154 mg) and used without purification in the next step. Boc₄N4Ac-O2Oc-OH (75 mg, 100 μmol, 1.2 eq), HATU (38 mg, 100 μmol, 1.2 eq) and DIPEA (68 μL, 400 μmol, 4 eq) were dissolved in 500 μL DMF. The mixture was stirred for 2 min to ensure pre-activation of the chelator-linker building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (90 mg, 83 μmol, 3BP-4089—described in Example 7a) in 2 mL DMF and 20 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 60 min all volatiles were removed in the vacuum and the remainder subjected to lyophilization. After performing the steps of ‘Cleavage method C’ the crude peptide was lyophilized and subsequently subjected to HPLC purification (20 to 45% B in 30 min—Kinetex) to yield 67.4 mg of the pure title compound (55% yield). HPLC: R_t=6.0 min. LC/TOF-MS: exact mass 1414.681 (calculated 1414.681). C₆₅H₁₀₂N₁₄O₁₅S₃(MW=1415.791).

Example 12: Synthesis of Hex-[Cys-(tMeBn(ReON4Ac-02Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4147)

To the solution of Hex-[Cys-(tMeBn(N4Ac-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (25 mg, 17.7 μmol, 1 eq) and Trichlorooxobis(triphenylphosphine)-rhenium(V) (14.7 mg, 17.7 μmol, 1 eq) in ethanol (3 mL) 10 μL DIPEA were added. The mixture was stirred overnight at 50° C. After reduction of the reaction solvent volume to approx. 0.5 mL an equal amount of water was added and the resulting solution subjected to HPLC purification (15 to 45% B in 30 min, eluents without TFA modifier—Kinetex) to yield 6.1 mg of the pure title compound (21% yield). HPLC: R_t=6.0 min. LC/TOF-MS: exact mass 1612.606 (calculated 1612.608). C₆₅H₉₈N₁₄O₁₆ReS₃(MW=1613.968).

Example 13: Synthesis of Hex-[Cys-(tMeBn(Bio-Ttds-Ttds-Ttds-Ttds-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4170)

Fmoc-Ttds-OH was loaded onto the trityl resin as described in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 100 μmol scale. Onto this resin the sequence (Bio-Ttds-Ttds-Ttds-Ttds-OH) was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’. After performing the steps of ‘Cleavage method B’ the remainder was lyophilized and subjected to HPLC purification to yield 116.8 mg (80%) of the purified intermediate product. Bio-Ttds-Ttds-Ttds-Ttds-OH (86 mg, 59 μmol, 1 eq), HATU (22.4 mg, 59 μmol, 1 eq) and DIPEA (20.5 μL, 120 μmol, 2 eq) were dissolved in 1 mL DMF. The mixture was stirred for 2 min to ensure pre-activation of the biotin-linker conjugate building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (64 mg, 59 μmol, 3BP-4089—described in Example 7a) in 2 mL DMF and 20 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 120 min all volatiles were removed in the vacuum and the remainder subjected to lyophilization. The remainder was subjected to HPLC purification (20 to 45% B in 30 min—Kinetex) to yield 27.46 mg of the pure title compound (18% yield). HPLC: R_t=7.3 min. LC/TOF-MS: exact mass 2518.274 (calculated 2518.273). C₁₁₇H₁₉₁N₁₉O₃₃S₄(MW=2520.145).

Example 14: Synthesis of Hex-[Cys-(tMeBn(DTPA-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4224)

Boc-O2Oc-OH (dicyclohexylamine salt) (20.5 mg, 46 μmol, 1 eq), Oxyma (9.8 mg, 69 μmol, 1.5 eq) and DIC (10.7 μL, 69 μmol) were dissolved in DMF and stirred for 5 min to ensure pre-activation of the linker building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (50 mg, 46 μmol, 3BP-4089—described in Example 7a) in 2 mL DMF and 20 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 4 hours another portion of Boc-O2Oc-OH (equal amounts as stated above) was pre-activated and added to the peptide reaction solution. The mixture was left to stir overnight. Then all volatiles were removed in vacuum and the remainder lyophilized from water/acetonitrile. The freeze-dried crude product was subject to ‘Cleavage method C’ to remove the Boc-protecting group and subsequently purified by preparative HPLC (15 to 45% B in 30 min—Kinetex) to yield 16.25 mg of the pure intermediate peptide Hex-[Cys(tMeBn(H-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (29% yield). For the next step DTPA(tBu)₄-OH (Diethylenetriamine-N,N,N″,N″-tetra-tert-butyl acetate-N′-acetic acid) (8.2 mg, 13.2 μmol, 1 eq), HATU (5 mg, 13.2 μmol, 1 eq) and DIPEA (4.6 μL, 26.4 μmol, 2 eq) were dissolved in 100 μL DMF. After stirring for 2 min to ensure pre-activation of the chelator building block this mixture was added to the solution of the 16.25 mg intermediate peptide (13.2 μmol) whose pH value had been adjusted to approximately 7.5-8 by addition of 5 μL DIPEA. After 180 minutes all volatiles were removed in the vacuum and the remainder subjected to HPLC purification (35 to 75% B in 30 min—Kinetex) to yield 12.76 mg (7 μmol) of the pure protected intermediate peptide Hex-[Cys(tMeBn(DTPA(tBu)₄-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (53% yield). The latter was subject to ‘Cleavage method C’, all volatiles removed in the vacuum and the remainder subjected to HPLC purification (15 to 45% B in 30 min—Kinetex) to yield 5.9 mg (3.7 μmol) of the pure title compound (53% yield—overall yield: 8%). HPLC: R_t=6.6 min. LC/TOF-MS: exact mass 1603.661 (calculated 1603.661). C₇₁H₁₀₅N₁₃O₂₃S₃(MW=1604.868).

Example 15: Synthesis of Hex-[Cys-(tMeBn(H-HYNIC-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4342)

Boc-HYNIC-OH (9.2 mg, 36 μmol, 1.3 eq), HATU (13.7 mg, 36 μmol, 1.3 eq) and DIPEA (12.2 μL, 72 μmol, 2.6 eq) were dissolved in 250 μL DMF. The mixture was stirred for 2 min to ensure pre-activation of the chelator-linker building block. Then this mixture was added to the solution of Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (30 mg, 27.8 μmol, 1 eq, 3BP-4089—described in Example 7a) in 400 μL DMF and 10 μL DIPEA which was added to adjust the pH value of the peptide solution to approximately 7.5-8. After 60 min all volatiles were removed in the vacuum, the remainder redissolved in DMSO and this solution directed to HPLC purification (25 to 55% B in 30 min—Kinetex) to yield 17.8 mg (13.5 μmol, 48.5%) of the intermediate protected peptide. The removal of the Boc-protecting group was achieved by treatment of the peptide with HCl (37%, 40 μL). The resulting mixture was dissolved with sodium acetate buffer (pH 4.5, 1.8 mL) and acetonitrile (0.2 mL) and the solution subjected to HPLC purification (20 to 50% B (0.02% formic acid in place of 0.1% TFA) in 30 min—Kinetex) to yield 1.15 mg (0.9 μmol) of the pure title compound (7% yield—overall yield: 3.4%). HPLC: R_t=6.9 min. LC/TOF-MS: exact mass 1218.505 (calculated 1218.502). C57H78N12O12S3 (MW=1219.503).

Example 16: Synthesis of Hex-[Cys-(tMeBn(NOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4310)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ NOTA(tBu)₂-OH (2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazonan-1-yl)acetic acid) was coupled. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 5.6 mg (4.1 μmol) of the pure title compound (4%). HPLC: R_t=6.8 min. LC/TOF-MS: exact mass 1368.592 (calculated 1368.592). C₆₃H₉₂N₁₂O₁₆S₃(MW=1369.676).

Example 17: Synthesis of Hex-[Cys-(tMeBn(DTPA2-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4309)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ DTPA2(tBu)₄-OH (3,6,9-tris(2-(tert-butoxy)-2-oxoethyl)-13,13-dimethyl-11-oxo-12-oxa-3,6,9-triazatetradecan-1-oic acid) was coupled. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 5.8 mg (3.9 μmol) of the pure title compound (3.9%). HPLC: R_t=6.5 min. LC/TOF-MS: exact mass 1458.587 (calculated 1458.587). C65H94N12O20S3 (MW=1459.711).

Example 18: Synthesis of Hex-[Cys-(tMeBn(NODAGA-O2Oc-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4251)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 50 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ consecutively Fmoc-O2Oc-OH and (R)-NODA-GA(tBu)₃-OH were coupled. 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 4.31 mg (2.7 μmol) of the pure title compound (5.4%). HPLC: R_t=6.7 min. LC/TOF-MS: exact mass 1585.687 (calculated 1585.687). C₇₂H₁₀₇N₁₃O₂₁S₃(MW=1586.896).

Example 19: Synthesis of Hex-[Cys-(tMeBn(NOTA-Ttds-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4344)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ consecutively Fmoc-Ttds-OH and NOTA(tBu)₂-OH (2-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazonan-1-yl)acetic acid) were coupled. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 10.1 mg (6.0 μmol) of the pure title compound (6%). HPLC: R_t=6.8 min. LC/TOF-MS: exact mass 1670.776 (calculated 1670.776). C₇₇H₁₁₈N₁₄O₂₁S₃(MW=1672.043).

Example 20: Synthesis of Hex-[Cys-(tMeBn(DTPA2-Ttds-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4352)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) from example 7b which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ Fmoc-Ttds-OH and DTPA2(tBu)₄-OH (3,6,9-tris(2-(tert-butoxy)-2-oxoethyl)-13,13-dimethyl-11-oxo-12-oxa-3,6,9-triazatetradecan-1-oic acid) were coupled. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 6.87 mg (3.9 μmol) of the pure title compound (3.9%). HPLC: R_t=6.7 min. LC/TOF-MS: exact mass 1760.771 (calculated 1760.771). C₇₉H₁₂₀N₁₄O₂₅S₃(MW=1762.078).

Example 21: Synthesis of Hex-[Cys-(tMeBn(H-SAc-Ser-Ser-Ser-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4301)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ Fmoc-Ser(tBu)-OH was coupled 3 times, followed by the coupling of Tritylmercapto acetic acid. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 5.25 mg (3.7 μmol) of the pure title compound (3.7%).

HPLC: R_t=6.8 min. LC/TOF-MS: exact mass 1418.553 (calculated 1418.538). C₆₂H₉₀N₁₂O₁₈S₄(MW=1419.714).

Example 22: Synthesis of Hex-[Cys-(tMeBn(H-Asp-Asp-Cys-Ttds-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4302)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) from example 7b which was used in a 100 μmol scale. According to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ Fmoc-Ttds-OH, Fmoc-Cys(Trt)-OH, and twice Fmoc-Asp(OtBu)-OH were coupled. After drying the resin was subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 45% B in 30 min—Kinetex) to yield 5.52 mg (3.2 μmol) of the pure title compound (3.2%). HPLC: R_t=6.8 min. LC/TOF-MS: exact mass 1718.705 (calculated 1718.706). C₇₆H₁₁₄N₁₄O₂₃S₄(MW=1720.066).

Example 23: Synthesis of Hex-[Cys-(tMeBn(DTPABzl-Glutar-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4366)

The starting point for the synthesis of the title compounds was the 3BP-4089 peptide resin from example 7b (Hex-[Cys(tMeBn(H-AET))-Pro-Pro-Thr(tBu)-Gln(Trt)-Phe-Cys]-O-WANG-Tentagel) which was used in a 100 μmol scale. Glutaric anhydride (57 mg, 0.5 mmol, 5 eq.) and DIPEA (165 μL, 1 mmol, 10 eq.) were dissolved in DMF (3 mL), the solution added to resin and the latter agitated for 1 hour. p-NH2-Bn-DTPA(OtBu)5 (S-2-(4-Aminobenzyl)-diethylenetriamine penta-tert-butyl acetate, 155 mg, 200 μmol, 2 eq.), Oxyma (27.2 mg, 200 μmol, 2 eq.), DIPEA (70 μL, 400 μmol, 4 eq.) and DIC (31 μL, 200 μmol, 2 eq.) were dissolved in DMF (1.7 mL), the solution added to the resin and the latter agitated for 90 minutes at 50° C. The addition of DIC was repeated and the agitation of the resin at 50° C. repeated for another 90 minutes. Thereafter another portion of DIC was added and the resin agitated at room temperature overnight. The next the DIC addition with subsequent agitation at 50° C. was repeated another 3 times. Then the resin was washed and subjected to ‘Cleavage method B’. The crude peptide was lyophilized and subsequently purified by preparative HPLC (20 to 40% B in 30 min—Kinetex) to yield 10.53 mg (6.3 μmol) of the pure title compound (6.3%). HPLC: R_t=7.0 min. LC/TOF-MS: exact mass 1677.688 (calculated 1677.676). C₇₇H₁₀₇N₁₃O₂₃S₃(MW=1678.948).

Example 24: Synthesis of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-AET](3BP-3654)

This synthesis was performed as the synthesis of 3BP-3554 described in Example 7a except for the fact that a commercially available pre-loaded aminoethanthiol trityl resin was used for the assembly of the linear peptide precursor Hex-Cys-Pro-Pro-Thr-Gln-Phe-AET. After performing all the steps described in Example 7 HPLC purification (15 to 45% B in 30 min—Kinetex) finally yielded 21.25 mg of the pure title compound (29.8% overall yield). HPLC: R_t=6.2 min. LC/TOF-MS: exact mass 1425.661 (calculated 1425.649). C₆₆H₉₉N₁₃O₁₆S₃(MW=1426.771).

Example 25: Synthesis of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cysol](3BP-3762)

This synthesis was performed as the synthesis of 3BP-3554 described in Example 7a except for the fact that Fmoc-Cysteinol(Trt)-OH was loaded onto the trityl resin. Differing from the description in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ this was achieved as follows: 50 μmol of trityl resin were swollen in THF and subsequently washed with dry THF (3 times). Then the resin was treated with a solution of Fmoc-Cysteinol(Trt)-OH (57 mg, 100 μmol, 2 eq) and pyridine (16.1 μl, 200 μmol, 4 eq) in dry THF (1 ml) for 20 hours at 60° C. After washing the resin thoroughly (THF, MeOH, DCM, DMF, 3 ml, 3×1 min) the linear peptide precursor Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cysol was assembled as described in the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’. After performing all the steps described in Example 7 HPLC purification (15 to 45% B in 30 min—Kinetex) finally yielded 7.8 mg of the pure title compound (10.7% overall yield). HPLC: R_t=5.9 min. LC/TOF-MS: exact mass 1455.666 (calculated 1455.660). C₆₇H₁₀₁N₁₃O₁₇S₃(MW=1456.797).

Example 26: Synthesis of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3407)

a) Synthesis of Intermediate Hex-[Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂by Two Different Cyclization Methods

The sequence (Hex-Cys-Pro-Pro-Thr-Gln-Phe-Cys-Asp-NH₂) of the peptide was assembled according to the ‘General procedures for Automated/Semi-automated Solid-Phase Synthesis’ in a 50 μmol scale on a Rink amide resin. After performing the steps of ‘Cleavage method B’ the crude peptide was lyophilized and cyclized by two alternative methods.

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 first 30 μl DIPEA and then 26.8 mg of 1,3,5-tris(bromomethyl)benzene (75 μmol, 1.5 eq compared to initial resin loading) were added. After stirring the solution for 45 minutes a solution of 43 mg piperazine (500 μmol, 10 eq compared to initial resin loading) in 200 μl of a 1:1 mixture of ethanol/acetonitrile was added. After 1 hour the solvents were removed in vacuo, 25 ml of a 1:1 mixture of acetonitrile and water (containing 50 μl TFA) was added and the solvents were removed by lyophilization. The remainder was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 15.3 mg (12.7 μmol) of the peptide intermediate Hex-Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(25.3%).

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 26.8 mg of 1,3,5-tris(bromomethyl)benzene (75 μmol, 1.5 eq compared to initial resin loading) were added. The solution was stirred for 1 hour and 43 mg piperazine (500 μmol, 10 eq compared to initial resin loading) were added. After 6 hours 100 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 17.2 mg (14.2 μmol) of the peptide intermediate Hex-Cys(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(28.4%).

Both cyclization methods perform similar and achieve comparable yields and similar purities.

b) Final Steps of Synthesis of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3407): DOTA-Coupling and Purification

To the solution of the intermediate (obtained by cyclization method B) in 200 μl DMSO 2.5 μl DIPEA were added to adjust the pH value to approximately 7.5-8. Then 16.3 mg of DOTA-NHS (21.4 μmol, 1.5 eq compared to the peptide intermediate) in 100 μl DMSO were added. During the course of the LC/TOF-MS monitored reaction 2.5 μl DIPEA was added 5 times to re-adjust the pH value to the starting value. After reaction completion the solution was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 19.1 mg (12.0 μmol) of the pure title compound (85%). HPLC: R_t=5.70 min. LC/TOF-MS: exact mass 1592.737 (calculated 1592.737). C₇₃H₁₀₈N₁₆O₂₀S₂(MW=1593.866).

Example 27: Synthesis of Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3476)

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 first 25 μl DIPEA and then a solution of 15.9 mg 1,3,5-tris(bromomethyl)benzene (60 μmol, 1.2 eq compared to initial resin loading) in 60 μl acetonitrile/ethanol 1:1 was added. The solution was stirred for 90 minutes and then 77 mg dithiothreitol (500 μmol, 10 eq compared to initial resin loading) was added. After stirring overnight the solvents were removed in vacuo and 30 ml of a 1:1 mixture of acetonitrile and water (containing 50 μl TFA) were added. The solvents were removed by lyophilization. The remainder was subjected to HPLC purification (15 to 40% B in 30 min—Kinetex) to yield 16.0 mg (14.4 μmol) of the pure title compound (28.8%). HPLC: R_t=7.36 min. LC/TOF-MS: exact mass 1108.476 (calculated 1108.472). C₅₂H₇₂N₁₀O₁₃S₂(MW=1109.320).

Cyclization Method B:

The lyophilized 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 a solution of 15.8 mg α,α′-dibromo-m-xylene (60 μmol, 1.2 eq compared to initial resin loading) in 0.5 ml acetonitrile was added. Upon completion of the cyclization reaction 50 μl TFA were added and the solvent removed by lyophilization. The remainder was subjected to HPLC purification (25 to 45% B in 30 min—Kinetex) to yield 16.9 mg (15.2 μmol) of the pure title compound (30.4%). HPLC: R_t=7.24 min. LC/TOF-MS: exact mass 1108.476 (calculated 1108.472). C₅₂H₉₂N₁₀O₁₃S₂(MW=1109.320).

Both cyclization methods (A and B) are similar effective in terms of yields and purity and are therefore both applicable.

Example 28: Preparation of DOTA-Transition Metal Complexes of Compounds of the Invention

A. General Procedure for the Preparation of a Peptide Comprising DOTA-Transition Metal-Complexes from Corresponding Peptides Comprising Uncomplexed DOTA

A 0.1 mM solution of the peptide comprised by uncomplexed DOTA in

- 0.4 M sodium acetate, pH=5 (Buffer A) (in case of Cu(II), Zn(II), In(III), Lu(III) or Ga(III) complexes) or
- 0.1 M ammonium acetate, pH=8 (Buffer B) (in case of Eu(III) complexes) was diluted with a solution 0.1 mM solution of the corresponding metal salt in water whereby the molar ratio of peptide to metal was adjusted to 1:3. The solution was stirred
- at 50° C. for 20 minutes (also referred to herein as Condition A) (in case of In(III), Lu(III), Ga(III), Zn(II) or Cu(II) complexes) or
- at room temperature overnight (also referred to herein as Condition B) (in case of Eu(III) complexes).

The solution was then applied to

- HPLC purification (also referred to herein as Purification A) or
- solid phase extraction (also referred to herein as Purification B).
- In case of solid phase extraction 250 mg Varian Bondesil-ENV was placed in a 15 ml polystyrene syringe, pre-washed with methanol (1×5 ml) and water (2×5 ml). Then the reaction solution was applied to the column. Thereafter elution was performed with water (2×5 ml—to remove excess salt), 5 ml of 50% ACN in water as first fraction and each of the next fractions were eluted with 5 ml of 50% ACN in water containing 0.10% TFA.

In either case (HPLC purification or solid phase extraction) fractions containing the pure product were pooled and freeze dried.

B. Indium-Complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3590)

The complex was prepared starting from 25 mg peptide 3BP-3407 (15.7 μmol) dissolved in Buffer A, diluted with a solution of InCl₃×4 H₂O which was treated with Condition A. In the purification step ‘Purification A’ was employed (15 to 40% B in 30 min—RLRP-S) to yield 18.24 mg of the pure title compound (68.1% yield). HPLC: R_t=5.6 min. LC/TOF-MS: exact mass 1702.622 (calculated 1702.617). C₇₃H₁₀₅InN₁₆O₂OS₂(MW=1705.663).

C. Gallium-Complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3592)

The complex was prepared starting from 25 mg peptide 3BP-3407 (15.7 μmol) dissolved in Buffer A, diluted with a solution of Ga(NO₃)₃×H₂O which was treated with Condition A. In the purification step ‘Purification A’ was employed (15 to 40% B in 30 min—RLRP-S) to yield 16.78 mg of the pure title compound (69.3% yield). HPLC:R_t=5.7 min. LC/TOF-MS: exact mass 1658.664 (calculated 1658.639). C₇₃H₁₀₅GaN₁₆O₂₀S₂(MW=1660.568).

D. Lutetium-Complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3591)

The complex was prepared starting from 25 mg peptide 3BP-3407 (15.7 μmol) dissolved in Buffer A, diluted with a solution of LuCl₃which was treated with Condition A. In the purification step ‘Purification A’ was employed (15 to 40% B in 30 min—RLRP-S) to yield 16.66 mg of the pure title compound (60.1% yield). HPLC: R_t=5.6 min. LC/TOF-MS: exact mass 1764.654 (calculated 1764.654). C₇₃H₁₀₅LuN₁₆O₂₀S₂(MW=1765.812).

E. Europium-Complex of Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH₂(3BP-3661)

The complex was prepared starting from 9.5 mg peptide (6 μmol) 3BP-3407 dissolved in Buffer B, diluted with a solution of EuCl₃×6H₂O which was treated with Condition B. In the purification step ‘Purification B’ was employed to yield 8.24 mg of the pure title compound (79.3% yield). HPLC: R_t=5.7 min. LC/TOF-MS: exact mass 1740.636 (calculated 1740.633). C₇₃H₁₀₅EuN₁₆O₂OS₂(MW=1742.809).

F. Indium-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3623) The complex was prepared starting from 6 mg peptide 3BP-3554 (4.1 μmol) dissolved in Buffer A, diluted with a solution of InCl₃×4 H₂O which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 5.26 mg of the pure title compound (81% yield). HPLC: R_t=5.8 min. LC/TOF-MS: exact mass 1579.524 (calculated 1579.520). C₆₇H₉₆InN₁₃O₁₈S₃(MW=1582.574).

G. Lutetium-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3624)

The complex was prepared starting from 6 mg peptide 3BP-3554 (4.1 μmol) dissolved in Buffer A, diluted with a solution of LuCl₃which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 5.5 mg of the pure title compound (82% yield). HPLC: R_t=5.9 min. LC/TOF-MS: exact mass 1641.560 (calculated 1641.557). C₆₇H₉₆LuN₁₃O₁₈S₃(MW=1642.723).

H. Gallium-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3949)

The complex was prepared starting from 7.9 mg peptide 3BP-3554 (5.4 μmol) dissolved in Buffer A, diluted with a solution of Ga(NO₃)₃×H₂O which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 4.2 mg of the pure title compound (51% yield). HPLC: R_t=6.6 min. LC/TOF-MS: exact mass 1535.543 (calculated 1535.541). C₆₇H₉₆GaN₁₃O₁₈S₃(MW=1537.479).

I. Europium-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-3662) The complex was prepared starting from 3.4 mg peptide 3BP-3554 (2.3 μmol) dissolved in Buffer B, diluted with a solution of EuCl₃×6 H₂O which was treated with Condition B. In the purification step ‘Purification B’ was employed to yield 3.1 mg of the pure title compound (83% yield). HPLC: R_t=5.9 min. LC/TOF-MS: exact mass 1617.541 (calculated 1617.536). C₆₇H₉₆EuN₁₃O₁₈S₃(MW=1619.721).

J. Copper(II)-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4293)

The complex was prepared starting from 18 mg peptide 3BP-3554 (12.2 μmol) dissolved in Buffer A, diluted with a solution of Cu(OAc)₂which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 16.5 mg of the pure title compound (88% yield). HPLC: R_t=6.5 min. LC/TOF-MS: exact mass 1530.553 (calculated 1530.553). C₆₇H₉₇CuN₁₃O₁₈S₃(MW=1532.310).

K. Zink-Complex of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4343)

The complex was prepared starting from 20 mg peptide 3BP-3554 (13.6 μmol) dissolved in Buffer A, diluted with a solution of ZnCl₂which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 16.1 mg of the pure title compound (77% yield). HPLC: R_t=6.4 min. LC/TOF-MS: exact mass 1531.553 (calculated 1531.553). C₆₇H₉₇N₁₃O₁₈S₃Zn (MW=1534.160).

L. Gallium-Complex of Hex-[Cys(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4184)

The complex was prepared starting from 7.4 mg peptide 3BP-4162 (5.1 μmol) dissolved in Buffer A, diluted with a solution of Ga(NO₃)₃×H₂O which was treated with Condition A. In the purification step ‘Purification B’ was employed to yield 6.3 mg of the pure title compound (80% yield). HPLC: R_t=6.5 min. LC/TOF-MS: exact mass 1506.515 (calculated 1506.515). C₆₆H₉₃GaN₁₂O₁₈S₃(MW=1508.438).

Example 29: Plasma Stability Assay

In order to determine the stability of selected compounds of the invention in human and mouse plasma, a plasma stability assay was carried out. Such plasma stability assay measures degradation of compounds of the present invention in blood plasma. This is an important characteristic of a compound as compounds, with the exception of pro-drugs, which rapidly degrade in plasma, generally show poor in vivo efficacy. The results show that those compounds are highly stable in human and mouse plasma. The stability is sufficient for the diagnostic, therapeutic and theragnostic use of these compounds according to the present invention.

The plasma stability samples were prepared by spiking 50 μl plasma aliquots (all K2EDTA) with 1 μl of a 0.5 mM compound stock solution in DMSO. After vortexing the samples were incubated in a Thermomixer at 37° C. for 0, 4 and 24 hours. After incubation the samples were stored on ice until further treatment. All samples were prepared in duplicates.

A suitable 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 as indicated in Table 8.

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

B) 150 μl of a zinc sulphate precipitation agent containing 78% 0.1 M zinc sulphate and 22% acetonitrile was added. After incubation at room temperature for 30 min the precipitate was separated by centrifugation. To 100 μl of the supernatant 10 μl of 1% formic acid was added followed by further incubation at 60° C. for 10 min to complete the formation of the zinc chelate, if the compound contains a free DOTA moiety.

The determination of the analyte in the clean sample solutions was performed on an Agilent 1290 UHPLC system coupled to an Agilent 6530 Q-TOF mass spectrometer. The chromatographic separation was carried out on a Phenomenex BioZen XB-C18 HPLC column (50×2 mm, 1.7 μm particle size) with 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 with a sampling rate of 2/sec.

From the mass spectrometric raw data the ion currents for the double or triple charged monoisotopic signal was extracted for both, the compound and the internal standard.

Quantitation was performed by external matrix calibration with internal standard using the integrated analyte signals.

Additionally, recovery was determined by spiking a pure plasma sample that only contained the internal standard after treatment with a certain amount of the compound.

Carry-over was evaluated by analysis of a blank sample (20% acetonitrile) after the highest calibration sample.

The results of this assay performed on some of the compounds according to the present invention are given in the following Table 8. The result is stated as “% intact compound remaining after 4 h or 24 h” and means that from the amount of material at the start of the experiment the stated percentage is detected as unchanged material at the end of the experiment by LC-MS quantification. Since all compounds are more than 50% intact after at least 4 h they are considered as stable enough for diagnostic and therapeutic applications.

TABLE 8

Results of the plasma stability assay

% intact compound remaining

Protein
after 4/24 h incubation

precipitation
Human
Mouse
Rat

Compound
method
plasma
plasma
plasma

3BP-2974
A

92%
(4 h)

3BP-2975
A

100%
(4 h)

3BP-2976
A

93%
(4 h)

3BP-3086
A

79%
(4 h)

3BP-3105
A

55%
(4 h)

3BP-3168
A

100%
(4 h)

3BP-3177
A

79%
(4 h)

3BP-3181
A

100%
(4 h)

3BP-3183
A

98%
(4 h)

3BP-3187
A

100%
(4 h)

3BP-3188
A

97%
(4 h)

3BP-3189
A

100%
(4 h)

3BP-3190
A

88%
(4 h)

3BP-3191
A

100%
(4 h)

3BP-3196
A

87%
(4 h)

3BP-3202
A

78%
(4 h)

3BP-3203
A

100%
(4 h)

3BP-3210
A

100%
(4 h)

3BP-3211
A

85%
(4 h)

3BP-3212
A

80%
(4 h)

3BP-3275
A

94%
(4 h)

3BP-3319
A

100%
(4 h)

3BP-3320
A

75%
(4 h)

3BP-3321
A

94%
(4 h)

3BP-3397
A
100% (24 h)
92%
(24 h)

3BP-3398
A
99% (24 h)
94%
(24 h)

3BP-3407
A
100% (24 h)
79%
(24 h)
100% (24 h)

3BP-3426
B

73%
(24 h)

3BP-3554
B
100% (24 h)
85%
(24 h)
100% (24 h)

3BP-3555
B

88%
(24 h)

3BP-3590
B
94% (24 h)
100%
(24 h)
100% (24 h)

3BP-3623
B
100% (24 h)
100%
(24 h)
100% (24 h)

3BP-3624
B
100% (24 h)
100%
(24 h)
100% (24 h)

Example 30: FACS Binding Assay

In order to determine binding of compounds according to the present invention to FAP-expressing cells, a competitive FACS binding assay was established.

FAP-expressing human WI-38 fibroblasts (ECACC) were cultured in EMEM including 15% fetal bovine serum, 2 mM L-Glutamine and 1% Non-essential amino acids. Cells were detached with Accutase (Biolegend, #BLD-423201) and washed in FACS buffer (PBS including 1% FBS). Cells were diluted in FACS buffer to a final concentration of 100.000 cells per ml and 200 μl of the cell suspension are transferred to a u-shaped non-binding 96-well plate (Greiner). Cells were washed in ice-cold FACS buffer and incubated with 3 nM of Cy5-labeled compound (H-Met-[Cys(3 MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-His-Phe-Arg-Asp-Ttds-Lys (Cy5 SO3)-NH2) in the presence of increasing concentrations of peptides at 4° C. for 1 hour. Cell were washed twice with FACS buffer and resuspended in 200 μl FACS buffer. Cells were analyzed in an Attune NxT flow cytometer. Median fluorescence intensities (Cy5 channel) was calculated by Attune NxT software and plotted against peptide concentrations. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. The results of this assay as well as the ones of the FAP protease activity assay as subject to Example 31 for each compound according to the present invention are presented in Table 9 (shown in Example 31). pIC50 category A stands for pIC50 values >8.0, category B for pIC50 values between 7.1 and 8.0, category C for pIC50 values between 6.1 and 7.0 and category D for pIC50 values ≤6.0.

Example 31: FAP Protease Activity Assay

In order to determine the inhibitory activity of compounds according to the present invention to FAP-expressing cells, a FRET-based FAP protease activity assay was established.

Recombinant human FAP (R&D systems, #3715-SE) was diluted in assay buffer (50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5) to a concentration of 3.6 nM. 25 μl of the FAP solution was mixed with 25 μl of a 3-fold serial dilution of the test compounds and incubated for 5 min in a white 96-well ProxiPlate (Perkin Elmer). As specific FAP substrate the FRET-peptide HiLyteFluor™ 488—VS(D-)P SQG K(QXL® 520)—NH2 was used (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. prior to use. Substrate cleavage and increase in fluorescence (excitation at 485 nm and emission at 538 nm) was measured in a kinetic mode for 5 minutes at 37° C. in a SPECTRAmax M5 plate reader. RFU/sec was calculated by SoftMax Pro software and plotted against peptide concentration. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. The results of this assay for each compound according to the present invention are given in Table 9. pIC50 category A stands for pIC50 values >8.0, category B for pIC50 values between 7.1 and 8.0, category C for pIC50 values between 6.1 and 7.0 and category D for pIC50 values ≤6.0.

As evident from Table 9, the compounds of the present invention show surprisingly superior results in both the FACS Binding assay and the FAP protease activity assay.

In addition to this one can easily find SAR-data which demonstrates that compounds with conjugated chelator are of very similar activity to compounds without chelator but similar peptide sequence. For instance, 3BP-3168 and 3BP-3169 possess chelator and linker at the C-terminus (DOTA-Ttds-Nle/Met) and are in the highest activity categories of pIC₅₀>8. Corresponding compounds without chelator and linker at the N-Terminus (3BP-2974 with N-terminal Hex-, 3BP-2975 with N-terminal Ac-Met and 3BP-2976 with N-terminal H-met) exhibit all similar activity compared to the chelator comprising compounds 3BP-3168 and 3BP-3169.

This means that the activity data from chelator free compounds is predictive for the activity of the chelator comprising compounds. This phenomenon is additionally also observed if the chelator is conjugated to the compounds of invention according to the other two specified possibilities. Examples for chelator attachment to the C-terminus compared to corresponding compounds without chelator show the same trends and are 3BP-3105 vs. 3BP-2974, 3BP-3395 or 3BP-3397 vs. 3BP-3476 and examples for the attachment of the chelator to Y, vs. corresponding compounds without chelator are 3BP-3407 vs. 3BP-3476 or 3BP-3426 vs. 3BP-3476.

TABLE 9

Compound ID, sequence, exact calculated mass, exact mass found, retention time in

minutes as determined by HPLC and pIC50 category of FACS binding and FAP activity assay

Exact
Exact

pIC50
pIC50

Mass
Mass
R_t
Category
Category

ID
Sequence
(calc)
(found)
(HPLC)
(FACS)
(Activity)

3BP-
H-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
2354.036
2354.046
5.51
A
A

2881
Asp-His-Phe-Arg-Asp-Ttds-Lys(Bio)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1664.712
1664.718
7.19
A
A

2974
His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.692
6.12
A
A

2975
Asp-His-Phe-Arg-Asp-NH2

3BP-
H-met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1697.679
1697.679
5.58
A
A

2976
Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
2065.903
2065.903
5.44
C
C

3088
Asp-His-Phe-Arg-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
2481.171
2481.171
6.78
A
A

3105
His-Phe-Arg-Asp-Ttds-Lys(DOTA)-NH2

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2368.087
2368.093
6.00
A
A

3168
Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Ttds-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-
2386.043
2386.050
5.74
A
A

3169
Phe-Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2083.859
2083.852
5.37
C
C

3170
Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Ttds-Phe-[Cys(3MeBn)-Pro-Pro-Thr-Glu-
2402.071
2402.075
6.00
B
B

3171
Phe-Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Ttds-Leu-[Cys(3MeBn)-Pro-Pro-Thr-Glu-
2368.087
2368.091
5.90
A
A

3172
Phe-Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Ttds-Glu-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2384.045
2384.049
5.19
B
B

3173
Cys]-Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1952.819
1952.822
4.86
C
C

3174
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.696
5.85
A
A

3175
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.693
6.20
C
C

3176
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-pro-Thr-Glu-Phe-Cys]-
1739.689
1739.689
5.85
B
B

3177
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-thr-Glu-Phe-Cys]-
1739.689
1739.692
5.61
B
B

3178
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-glu-Phe-Cys]-
1739.689
1739.692
5.98
B
C

3179
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-phe-Cys]-
1739.689
1739.693
5.97
C
C

3180
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-cys]-
1739.689
1739.695
6.24
B
B

3181
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-cys]-
1739.689
1739.695
6.12
B
B

3182
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.694
6.00
B
B

3183
asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.695
6.34
A
A

3187
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1753.705
1753.716
5.87
A
A

3188
Asp-His-Nmf-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1751.689
1751.697
5.71
A
A

3189
Asp-His-Tic-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1751.689
1751.701
6.38
A
A

3190
Asp-His-Aic-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1740.685
1740.696
5.00
A
A

3191
Asp-His-Ppa-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1740.685
1740.696
5.08
A
A

3192
Asp-His-Mpa-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Thi-Cys]-
1745.646
1745.650
6.03
A
A

3193
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1695.700
1695.703
6.16
B
B

3194
Ala-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1673.668
1673.670
6.97
A
B

3195
Asp-Ala-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1663.658
1663.661
5.43
A
A

3196
Asp-His-Ala-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1654.625
1654.634
6.51
C
C

3197
Asp-His-Phe-Ala-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1695.700
1695.711
6.30
A
A

3198
Asp-His-Phe-Arg-Ala-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1468.561
1468.570
6.64
C
B

3199
Asp-His-Phe-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1624.663
1624.669
6.31
A
A

3200
Asp-His-Phe-Arg-NH2

3BP-
Ac-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1608.649
1608.659
5.82
A
A

3202
His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1753.705
1753.705
6.47
A
A

3203
Asp-His-Amf-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Aib-Pro-Thr-Glu-Phe-Cys]-
1727.689
1727.700
6.51
B
B

3204
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.694
6.10
A
A

3210
Asp-his-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.692
5.77
A
A

3211
Asp-His-phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.692
5.88
A
A

3212
Asp-His-Phe-arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1739.689
1739.693
6.16
A
A

3213
Asp-His-Phe-Arg-asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Gly-Pro-Thr-Glu-Phe-Cys]-
1699.658
1699.662
5.71
A
A

3214
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-ala-Pro-Thr-Glu-Phe-Cys]-
1713.674
1713.677
5.99
C
C

3215
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Oic-Pro-Thr-Glu-Phe-Cys]-
1793.736
1793.739
6.91
C
C

3216
Asp-His-Phe-Arg-Asp-NH2

3BP-
Ac-Met-[Cys(3MeBn)-Pro-Oic-Thr-Glu-Phe-Cys]-
1793.736
1793.740
6.83
C
C

3217
Asp-His-Phe-Arg-Asp-NH2

3BP-
DOTA-Bal-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
2037.871
2037.875
4.71
B
B

3264
Asp-His-Nmf-Arg-Asp-NH2

3BP-
DOTA-Inp-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]
2077.903
2077.902
4.83
C
C

3265
Asp-His-Nmf-Arg-Asp-NH2

3BP-
DOTA-Ahx-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2079.918
2079.923
4.95
C
C

3266
Cys]-Asp-His-Nmf-Arg-Asp-NH2

3BP-
DOTA-O2Oc-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2111.908
2111.909
4.93
C
C

3267
Cys]-Asp-His-Nmf-Arg-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
2380.160
2380.167
6.57
A
A

3275
His-Nmf-Arg-Ttds-Lys(DOTA)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
2366.144
2366.151
6.47
A
A

3276
His-phe-Arg-Ttds-Lys(DOTA)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
2367.139
2367.150
5.79
A
A

3277
His-Ppa-arg-Ttds-Lys(DOTA)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-NH2
994.429
994.432
7.59
C
B

3287

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1109.456
1109.458
7.42
A
A

3288
NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1265.557
1265.562
7.30
A
A

3299
Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1350.610
1350.611
7.29
A
A

3300
Gab-Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1398.610
1398.616
7.38
A
A

3301
Pamb-Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1390.641
1390.641
7.21
A
A

3302
Omp-Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-
1283.583
1283.588
7.68
B
A

3303
Pamb-Arg-NH2

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
1697.804
1697.810
5.81
B
B

3319
Cys]-NH2

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
1812.831
1812.841
5.75
B
A

3320
Cys]-Asp-NH2

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-
2101.985
2101.993
5.49
A
A

3321
Cys]-Asp-Pamb-Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1398.610
1398.614
7.40
A
A

3324
Mamb-Arg-NH2

3BP-
Hex-[Cys(3MeBn)-Gly-Pro-Thr-Glu-Phe-Cys]-NH2
954.398
954.402
7.32
C
B

3345

3BP-
Hex-[Cys(3MeBn)-Ala-Pro-Thr-Glu-Phe-Cys]-NH2
968.414
968.415
7.46
D
C

3346

3BP-
Hex-[Cys(3MeBn)-Nmg-Pro-Thr-Glu-Phe-Cys]-NH2
968.414
968.416
7.37
C
B

3347

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Ala-Phe-Cys]-NH2
936.424
936.426
7.75
D
D

3348

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-NH2
993.445
993.449
7.41
B
A

3349

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Ala-Cys]-NH2
918.398
918.398
6.49
D
C

3350

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Pen]-NH2
1022.461
1022.463
7.84
D
C

3351

3BP-
Hex-[Cys(3MeBn)-Pro-4Tfp-Thr-Glu-Phe-Cys]-NH2
1012.420
1012.422
7.72
C
B

3352

3BP-
Hex-[Cys(3MeBn)-Pro-Eay-Thr-Glu-Phe-Cys]-NH2
1070.461
1070.464
9.10
D
C

3353

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Ala-Glu-Phe-Cys]-NH2
964.419
964.418
7.63
D
D

3354

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Opc-Glu-Phe-Cys]-NH2
1113.478
1113.480
7.63
D
C

3355

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Moo-Phe-Cys]-NH2
1028.417
1028.419
7.75
D
D

3356

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Nme-Phe-Cys]-NH2
1008.445
1008.448
7.82
D
D

3357

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Nmf-Cys]-NH2
1008.445
1008.445
8.12
D
C

3358

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Tic-Cys]-NH2
1006.429
1006.431
8.11
D
C

3359

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Nphe-Cys]-NH2
994.429
994.432
7.85
D
D

3360

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-1Ni-Cys]-NH2
1044.445
1044.448
8.50
B
B

3361

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-2Ni-Cys]-NH2
1044.445
1044.449
8.46
C
C

3362

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Bip-Cys]-NH2
1070.461
1070.464
9.04
D
D

3363

3BP-
Hex-[Cys(3MeBn)-Pro-4Dfp-Thr-Glu-Phe-Cys]-NH2
1030.410
1030.414
8.01
D
C

3365

3BP-
Hex-[Cys(3MeBn)-Pro-Hyp-Thr-Glu-Phe-Cys]-NH2
1010.424
1010.428
7.18
B
B

3366

3BP-
Hex-[Cys(3MeBn)-Pro-Tap-Thr-Glu-Phe-Cys]-NH2
1009.440
1009.445
6.87
D
C

3367

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Ocf-Cys]-NH2
1028.390
1028.394
7.95
C
B

3368

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Pcf-Cys]-NH2
1028.390
1028.394
8.14
C
C

3369

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH
995.413
995.417
7.79
B
B

3370

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Bal-
1066.450
1066.453
7.58
A
B

3371
OH

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Orn(Ac)-Glu-Phe-Cys]-
1049.471
1049.475
7.33
D
C

3372
NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1924.931
1924.943
6.60
A
A

3395
Ttds-Lys(DOTA)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-Asp-
1925.915
1925.916
6.73
A
A

3396
Ttds-Lys(DOTA)-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1522.720
1522.714
6.69
A
A

3397
Bhk(DOTA)-OH

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1768.842
1768.842
5.72
A
A

3398
Cys]-Bal-OH

3BP-
DOTA-Ttds-Hci-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1826.858
1826.858
4.78
D
D

3399
Cys]-Bal-OH

3BP-
DOTA-Ttds-Hgl-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1796.873
1796.873
6.58
D
B

3400
Cys]-Bal-OH

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1811.847
1811.855
5.62
A
A

3401
Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1579.741
1579.742
6.61
A
A

3403
Ape-NH-DOTA′

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1881.926
1881.933
6.73
A
A

3404
Ttds-Ape-NH-DOTA′

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-
1592.737
1592.737
5.70
A
A

3407
Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Trp-Cys]-NH2
1032.456
1032.457
7.58
B
B

3408

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Otf-Cys]-NH2
1061.433
1061.437
8.08
B
A

3409

3BP-
Oct-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1136.503
1136.508
8.46
B
B

3417
NH2

3BP-
Phb-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1156.472
1156.475
7.73
C
C

3418
NH2

3BP-
[3MeBn-Spa-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2
995.388
995.392
6.05
C
C

3419

3BP-
PentylNH-urea-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1123.483
1123.485
7.22
B
A

3425
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1583.682
1583.692
5.87
A
A

3426
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu(NH-Apr-
1551.710
1551.713
6.57
D
D

3472
DOTA′)-Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Glu(NH-Apr-O2Oc-
1696.784
1696.793
6.64
C
C

3473
DOTA′)-Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-
1108.472
1108.476
7.24
A
A

3476
NH2

3BP-
Hex-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1824.904
1824.922
6.66
A
A

3489
Bhk(DOTA-Ttds)-OH

3BP-
Pentyl-SO2-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1144.439
1144.442
7.79
A
A

3514
Cys]-Asp-NH2

3BP-
Hex-[Cys(2Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2
1109.467
1109.469
5.54
A
A

3518

3BP-
Hex-[Cys(3Lut)-Pro-Pro-Thr-Gln-Phe-Cys]-Asp-NH2
1109.467
1109.469
5.27
A
A

3519

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.639
1469.640
5.89
A
A

3554
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-Phe-
1478.694
1478.699
5.37
B
A

3555
Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-NH))-Pro-Pro-Thr-Gln-Phe-
1523.679
1523.669
5.71
B
B

3556
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(InDOTA-PP))-Pro-Pro-Thr-Gln-
1702.617
1702.622
5.59
A
A

3590
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(LuDOTA-PP))-Pro-Pro-Thr-Gln-
1764.654
1764.654
5.65
A
A

3591
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(GaDOTA-PP))-Pro-Pro-Thr-Gln-
1658.639
1658.644
5.75
A
A

3592
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-
1579.520
1579.524
5.75
A
A

3623
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-Gln-
1641.557
1641.560
5.81
A
A

3624
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-1Ni-
1519.655
1519.667
5.64
A
A

3650
Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1540.676
1540.686
5.81
A
A

3651
Phe-Cys]-Bal-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1468.655
1468.667
5.85
A
A

3652
Phe-Cys]-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Glu-
1469.639
1469.639
5.96
A
A

3653
Phe-Cys]-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1425.649
1425.661
6.16
A
A

3654
Phe-AET]

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1526.661
1526.665
5.88
A
A

3656
Phe-Cys]-Gly-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1554.692
1554.697
5.98
A
A

3657
Phe-Cys]-Gab-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1556.671
1556.670
5.78
A
A

3658
Phe-Cys]-Ser-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1540.676
1540.682
5.88
A
A

3659
Phe-Cys]-Nmg-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gin-
1630.723
1630.728
6.85
A
A

3660
Phe-Cys]-Bhf-OH

3BP-
Hex-[Cys(tMeBn(EuDOTA-PP))-Pro-Pro-Thr-Gln-
1740.633
1740.636
5.72
A
A

3661
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(EuDOTA-AET))-Pro-Pro-Thr-Gln-
1617.540
1617.541
5.83
A
A

3662
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1470.635
1470.638
4.84
A
A

3664
Mpa-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1584.666
1584.666
5.83
A
A

3665
Phe-Cys]-Asp-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-
1443.624
1443.624
5.84
A
A

3678
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Hyp-Thr-Gln-
1485.634
1485.645
5.69
A
A

3679
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Otf-
1537.627
1537.626
6.46
A
A

3680
Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1583.682
1583.697
5.84
A
A

3681
Phe-Cys]-asp-NH2

3BP-
Ac-met-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1544.617
1544.633
4.97
B
B

3682
Phe-Cys]-OH

3BP-
Pentyl-SO2-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-
1505.606
1505.610
6.23
A
A

3690
Gln-Phe-Cys]-OH

3BP-
Pentyl-SO2-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-
1506.590
1506.593
6.40
A
B

3691
Glu-Phe-Cys]-OH

3BP-
Pentyl-SO2-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-
1628.704
1628.706
5.99
A
A

3692
Gln-Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(Cy5SO3-PP))-Pro-Pro-Thr-Gln-
1793.851
1793.850
8.94

B

3693
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(Cy5SO3-AET))-Pro-Pro-Thr-Gln-
1670.754
1670.752
9.58

C

3694
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-
1578.536
1578.539
5.60
A
A

3712
Phe-Cys]-NH2

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-
1535.530
1535.533
6.01
A
A

3713
Phe-AET]

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-
1636.541
1636.546
5.60
A
A

3714
Phe-Cys]-Gly-OH

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-Gln-
1650.557
1650.569
5.64
A
A

3715
Phe-Cys]-Nmg-OH

3BP-
Hex-[Cys(tMeBn(InDOTA-AET))-Nmg-Pro-Thr-Gln-
1553.504
1553.517
5.69
A
A

3716
Phe-Cys]-OH

3BP-
Pentyl-SO2-[Cys(tMeBn(InDOTA-PP))-Pro-Pro-Thr-
1738.584
1738.588
5.97
A
A

3717
Gln-Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1539.692
1539.691
5.66
A
A

3736
Phe-Cys]-Bal-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1539.692
1539.690
5.70
A
A

3737
Phe-Cys]-Nmg-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Ala-Phe-
1535.715
1535.721
5.88
C
C

3739
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-Gln-
1442.640
1442.640
5.66
A
A

3744
Phe-Cys]-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Ala-Gln-Phe-
1562.726
1562.732
5.44
C
C

3745
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Ala-Ala-Phe-
1505.705
1505.705
5.62
D
C

3746
Cys]-Asp-NH2

3BP-
Hex-[Cys(3MeBn)-Nlys-Pro-Thr-Gln-Phe-Cys]-NH2
1024.487
1024.490
6.28
D
D

3747

3BP-
Hex-[Cys(3MeBn)-Nphe-Pro-Thr-Gln-Phe-Cys]-NH2
1043.461
1043.462
8.64
D
D

3748

3BP-
Hex-[Cys(3MeBn)-Nleu-Pro-Thr-Gln-Phe-Cys]-NH2
1009.477
1009.479
8.32
D
D

3749

3BP-
H-Ahx-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1008.456
1008.456
4.77
D
D

3759
NH2

3BP-
H-Ava-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
994.440
994.441
4.66
D
D

3760
NH2

3BP-
H-Gab-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
980.425
980.425
4.60
D
D

3761
NH2

3BP-
4Pya-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-NH2
1014.409
1014.415
4.74
D
C

3762

3BP-
Ac-Hse-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1038.430
1038.430
4.83
D
C

3763
NH2

3BP-
Ac-Aad-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1080.441
1080.440
5.11
D
D

3764
NH2

3BP-
HO-Glutar-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1009.404
1009.403
5.33
C
C

3765
NH2

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1455.660
1455.666
5.91
A
A

3767
Phe-Cysol]

3BP-
Hex-[Cys(tMeBn(InDOTA-PP))-Pro-Pro-Thr-Gln-
1588.574
1588.579
5.30
A
A

3770
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Nmg-Pro-Thr-Gln-
1452.678
1452.678
5.12
A
A

3771
Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Thr-Pro-Phe-Gln-
1592.737
1592.759
5.35
D
D

3854
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Phe-Gln-Thr-Pro-Pro-
1592.737
1592.737
5.67
D
D

3855
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Thr-Gln-Pro-Phe-Pro-
1592.737
1592.749
5.10
D
D

3856
Cys]-Asp-NH2

3BP-
Hex-[Cys(tMeBn(DOTA-PP))-Pro-Gln-Phe-Pro-Thr-
1592.737
1592.737
5.37
C
C

3857
Cys]-Asp-NH2

3BP-
H2NSO2-But-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1044.387
1044.401
5.18
D
D

3860
Cys]-NH2

3BP-
Hex-[Cys(tMeBn(GaDOTA-AET))-Pro-Pro-Thr-Gln-
1535.541
1535.541
6.58
A
A

3949
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-O2Oc-PP))-Pro-Pro-Thr-Gln-
1351.631
1351.648
6.1
A
A

3967
Phe-Cys]-Asp-NH2

3BP-
H-Ahx-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Gln-
1538.751
1538.758
6.3
A
A

3980
Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys-(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-
1197.502
1197.508
6.7
B
A

3981
Cys]-Asp-NH2

3BP-
Hex-[Cys-(tMeBn(H-O2Oc-AET))-Pro-Pro-Thr-Gln-
1342.576
1342.578
6.5
A
A

4003
Phe-Cys]-Asp-NH2

3BP-
H-Ahx-Ttds-Nle-[Cys-(tMeBn(DOTA-PP))-Pro-Pro-
2023.016
2023.029
5.2
B
A

4004
Thr-Gln-Phe-Cys]-Asp-NH2

3BP-
Hex-[Cys-(tMeBn(N4Ac-AET))-Pro-Pro-Thr-Gln-
1269.607
1269.612
6.0
A
A

4063
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(N4Ac-O2Oc-AET))-Pro-Pro-Thr-
1414.681
1414.691
6.0
A
A

4088
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-
1083.459
1083.472
6.9
C
B

4089
Cys]-OH

3BP-
Hex-[D-Cys-(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.639
1469.646
6.3
A
A

4109
Phe-Cys]-OH

3BP-
Hex-[D-Cys-(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.639
1469.647
6.6
B
B

4110
Phe-D-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.639
1469.646
6.0
B
B

4111
Phe-D-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(ReON4Ac-O2Oc-AET))-Pro-Pro-
1612.606
1612.620
6.6
A
A

4147
Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(ReON4Ac-AET))-Pro-Pro-Thr-
1467.532
1467.543
6.8
A
A

4148
Gln-Phe-Cys]-OH

3BP-
N4Ac-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-Phe-
1498.756
1498.765
5.8
A
A

4161
Cys]-OH

3BP-
Hex-[Cys-(tMeBn(NODAGA-AET))-Pro-Pro-Thr-Gln-
1440.613
1440.623
6.8
A
A

4162
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(N4Ac-PP))-Pro-Pro-Thr-Gln-Phe-
1278.662
1278.669
5.5
A
A

4168
Cys]-OH

3BP-
Hex-[Cys-(tMeBn(N4Ac-O2Oc-PP))-Pro-Pro-Thr-
1423.736
1423.741
5.4
B
A

4169
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(Bio-Ttds-Ttds-Ttds-Ttds-AET))-
2518.273
2518.291
7.3
B
B

4170
Pro-Pro-Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-PP))-Pro-Pro-Thr-Gln-Phe-
1092.514
1092.524
5.8
B
B

4181
Cys]-OH

3BP-
Hex-[Cys(tMeBn(ATTO488-AET))-Pro-Pro-Thr-Gln-
1654.531
1654.530
6.9
B
B

4182
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(GaNODAGA-AET))-Pro-Pro-Thr-
1506.515
1506.522
6.6
A
A

4184
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH
994.429
994.431
7.9
B
B

4186

3BP-
Hex-[Cys-(tMeBn(DTPA-AET))-Pro-Pro-Thr-Gln-
1458.587
1458.594
6.5
B
B

4214
Phe-Cys]-OH

3BP-
N4Ac-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Gln-Phe-
1497.772
1497.780
5.7
B
A

4219
Cys]-OH

3BP-
N4Ac-APAc-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1336.667
1336.674
5.4
D
D

4220
Phe-Cys]-OH

3BP-
N4Ac-PEG6-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1531.767
1531.779
5.9
B
B

4221
Phe-Cys]-OH

3BP-
N4Ac-Glu-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1627.799
1627.810
5.7
B
B

4222
Phe-Cys]-OH

3BP-
N4Ac-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-Ppa-
1499.752
1499.768
4.6
C
C

4223
Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DTPA-O2Oc-AET))-Pro-Pro-Thr-
1603.661
1603.656
6.6
B
B

4224
Gln-Phe-Cys]-OH

3BP-
N4Ac-O2Oc-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1341.646
1341.642
5.4
D
C

4228
Phe-Cys]-OH

3BP-
DTPA-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1687.736
1687.749
6.4
C
B

4229
Phe-Cys]-OH

3BP-
N4Ac-gGlu-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-
1325.615
1325.610
5.5
D
D

4230
Phe-Cys]-OH

3BP-
N4Ac-Ttds-Glu(AGLU′)-Nle-[Cys-(3MeBn)-Pro-Pro-
1790.883
1790.909
5.4
D
D

4231
Thr-Glu-Phe-Cys]-OH

3BP-
N4Ac-Ttds-Glu-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-Phe-
1514.715
1514.715
5.0
C
D

4233
Cys]-OH

3BP-
N4Ac-Efa-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Glu-Phe-
1430.640
1430.643
5.7
B
B

4243
Cys]-OH

3BP-
N4Ac-gGlu-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-Gln-
1324.631
1324.635
5.4
D
D

4244
Phe-Cys]-OH

3BP-
N4Ac-gGlu-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-
1626.815
1626.821
5.7
B
B

4245
Gln-Phe-Cys]-OH

3BP-
N4Ac-Glu(AGLU′)-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-
1789.899
1789.901
5.5
B
B

4246
Thr-Gln-Phe-Cys]-OH

3BP-
N4Ac-gGlu-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-Thr-
1627.799
1627.805
5.9
B
B

4247
Glu-Phe-Cys]-OH

3BP-
N4Ac-Ttds-Glu(AGLU′)-Nle-[Cys-(3MeBn)-Pro-Pro-
1789.899
1789.895
5.3
D
D

4248
Thr-Gln-Phe-Cys]-OH

3BP-
N4Ac-Glu(AGLU′)-Ttds-Nle-[Cys-(3MeBn)-Pro-Pro-
1790.883
1790.909
5.7
B
B

4249
Thr-Glu-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DOTA-AET))-Pro-Pro-Thr-Glu-
1470.623
1470.626
6.4
A
B

4250
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(NODAGA-O2Oc-AET))-Pro-Pro-
1585.687
1585.689
6.7
A
A

4251
Thr-Gln-Phe-Cys]-OH

3BP-
N4Ac-Glu(AGLU′)-Glu(AGLU′)-Ttds-Nle-[Cys-
2082.026
2082.030
5.6
C
B

4265
(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-OH

3BP-
N4Ac-Glu(AGLU′)-Glu(AGLU′)-Ttds-Nle-[Cys-
2083.010
2083.013
5.6
B
B

4266
(3MeBn)-Pro-Pro-Thr-Glu-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(CuDOTA-AET))-Pro-Pro-Thr-Gln-
1530.553
1530.562
6.5
A
A

4293
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(N4Ac-Ttds-AET))-Pro-Pro-Thr-
1571.791
1571.807
5.9
A
A

4299
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(N4Ac-PEG6-AET))-Pro-Pro-Thr-
1604.802
1604.816
6.1
B
A

4300
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-SAc-Ser-Ser-Ser-AET))-Pro-
1418.538
1418.553
6.8
B
B

4301
Pro-Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-Asp-Asp-Cys-Ttds-AET))-Pro-
1718.706
1718.723
6.8
C
B

4302
Pro-Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-Asp-Asp-Cys-AET))-Pro-Pro-
1416.522
1416.536
6.6
C
B

4303
Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-SAc-Ser-Ser-Ser-Ttds-AET))-
1720.722
1720.730
6.9
B
B

4308
Pro-Pro-Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DTPA2-AET))-Pro-Pro-Thr-Gln-
1458.587
1458.597
6.5
B
B

4309
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(NOTA-AET))-Pro-Pro-Thr-Gln-
1368.592
1368.600
6.7
A
A

4310
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(H-HYNIC-AET))-Pro-Pro-Thr-Gln-
1218.502
1218.505
6.9
B
A

4342
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(ZnDOTA-AET))-Pro-Pro-Thr-Gln-
1531.553
1531.558
6.4
A
A

4343
Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(NOTA-Ttds-AET))-Pro-Pro-Thr-
1670.776
1670.777
6.8
A
A

4344
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DTPA2-Ttds-AET))-Pro-Pro-Thr-
1760.771
1760.773
6.7
C
B

4352
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DTPA2-PEG6-AET))-Pro-Pro-Thr-
1793.781
1793.786
6.8
B
B

4353
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys-(tMeBn(DTPABzl-Glutar-AET))-Pro-Pro-
1677.676
1677.688
7.0
C
C

4366
Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-Gln-
2943.026
2943.056
5.6
A
A

4372
Phe-Cys]-Asp-Gab-Arg-Ttds-Lys(AF488)-NH2

3BP-
Hex-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-Gln-
3547.394
3547.431
5.7
B
A

4373
Phe-Cys]-Asp-Gab-Arg-Ttds-Ttds-Ttds-Lys(AF488)-

NH2

3BP-
Hex-[Cys-(tMeBn(H-HYNIC-Ttds-AET))-Pro-Pro-
1520.687
1520.685
6.8
A
A

4376
Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[C(tMeBn(PCTA-AET))-Pro-Pro-Thr-Gln-Phe-
1445.618
1445.635
6.5
A
A

4379
Cys]-OH

3BP-
Hex-[C(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-
1560.579
1560.596
6.3
A
A

4380
Cys]-OH

3BP-
Hex-[C(tMeBn(HBED-AET))-Pro-Pro-Thr-Gln-Phe-
1597.654
1597.669
7.4
B
B

4381
Cys]-OH

3BP-
Hex-[C(tMeBn(DATA-AET))-Pro-Pro-Thr-Gln-Phe-
1468.644
1468.657
7.0
B
B

4382
Cys]-OH

3BP-
Hex-[Cys(tMeBn(HBED-PEG6-AET))-Pro-Pro-Thr-
1932.849
1932.888
7.4
B
B

4383
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DATA-Ttds-AET))-Pro-Pro-Thr-
1770.828
1770.836
7.1
B
B

4384
Gln-Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(NOPO-Ttds-AET))-Pro-Pro-Thr-
1862.763
1862.785
6.5
B
B

4385
Gln-Phe-Cys]-OH

3BP-
DOTA-Ttds-Nle-[Cys(tMeBn(DOTA-AET))-Pro-Pro-
2173.015
2173.023
5.1
B
B

4386
Thr-Gln-Phe-Cys]-OH

3BP-
Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
2400.141
2400.173
5.8
A
A

4391
Phe-Cys]-Asp-Ttds-Lys(DOTA)-NH2

3BP-
DOTA-Ttds-Nle-[Cys(tMeBn(DOTA-AET))-Pro-Pro-
3013.517
3103.535
5.0
B
B

4392
Thr-Gln-Phe-Cys]-Asp-Ttds-Lys(DOTA)-NH2

3BP-
DOTA-Ttds-Nle-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-
2628.307
2628.327
5.7
A
A

4393
Cys]-Asp-Ttds-Lys(DOTA)-NH2

3BP-
iHex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.639
1469.643
6.0
A
A

3907
Phe-Cys]-OH

3BP-
Chex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1481.639
1481.644
6.0
C
C

3908
Phe-Cys]-OH

3BP-
Cp-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-
1467.6237
1467.626
5.7
B
B

3909
Cys]-OH

3BP-
Pent-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1471.619
1471.619
5.4
A
A

3910
Phe-Cys]-OH

3BP-
Rth-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-
1469.603
1469.607
4.9
B
B

3911
Cys]-OH

3BP-
Pyn-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1451.592
1451.596
5.1
B
C

3912
Phe-Cys]-OH

3BP-
Hyn-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1465.608
1465.611
5.3
B
B

3913
Phe-Cys]-OH

3BP-
Peet-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1453.608
1453.611
5.3
B
B

3914
Phe-Cys]-OH

3BP-
Alloc-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1455.587
1455.591
5.5
B
B

3915
Phe-Cys]-OH

3BP-
Bulloc-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1469.603
1469.607
5.9
A
B

3916
Phe-Cys]-OH

3BP-
Cpentyl-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-
1482.635
1482.638
5.6
B
B

3917
Thr-Gln-Phe-Cys]-OH

3BP-
EtOPr-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1471.619
1471.620
5.0
A
A

3918
Phe-Cys]-OH

3BP-
Fur-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-
1469.603
1469.617
4.8
C
C

3919
Cys]-OH

3BP-
Sth-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-
1469.603
1469.620
5.0
B
B

3936
Cys]-OH

3BP-
MeOBut-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1471.619
1471.635
4.9
A
B

3937
Phe-Cys]-OH

3BP-
PrOAc-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-
1471.619
1471.635
5.5
A
A

3938
Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-
1470.635
1470.638
5.8
A
A

3940
Gln-Phe-Cys]-OH

3BP-
nBu-COyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-
1471.619
1471.623
6.2
A
A

3941
Gln-Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(LuDOTA-AET))-Pro-Pro-Thr-
1642.552
1642.548
5.8
A
A

4425
Gln-Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(InDOTA-AET))-Pro-Pro-Thr-
1582.515
1582.513
5.7
A
A

4426
Gln-Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(N4Ac-PP))-Pro-Pro-Thr-Gln-
1279.657
1279.666
5.0
A
A

4533
Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(N4Ac-AET))-Pro-Pro-Thr-
1270.603
1270.602
5.5
A
A

4534
Gln-Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-
1469.651
1469.699
5.7
A
A

4560
Gln-Phe-Cys]-NH2

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-
1479.689
1479.731
5.2
A
A

4564
Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-PP))-Pro-Pro-Thr-Gln-
1478.705
1478.733
5.3
A
A

4565
Phe-Cys]-NH2

3BP-
nBu-CAyl-[Cys(3MeBn)-Pro-Pro-Thr-Gln-Phe-Cys]-
1323.683
1323.705
5.8
A
A

4589
Bhk(N4Ac)-OH

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-
1444.619
1444.645
5.7
A
A

4607
Gln-Phe-Cys]-OH

3BP-
nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Nmg-Pro-Thr-
1443.635
1443.657
5.7
A
A

4621
Gln-Phe-Cys]-NH2

Example 32: Surface Plasmon Resonance Assay

Surface plasmon resonance studies were performed using a Biacore™ T200 SPR system. Briefly, polarized light is directed towards a gold-labeled sensor surface, and minimum intensity reflected light is detected. The angle of reflected light changes as molecules bind and dissociate. The gold-labeled sensor surface is loaded with FAP antibodies bearing FAP target proteins, whereby antibody binding does not occur at the substrate-binding site of FAP. Test compounds are contacted with the loaded surface, and a real-time interaction profile with the FAP ligand is recorded in a sensorgram. In real-time, the association and dissociation of a binding interaction is measured, enabling calculation of association and dissociation rate constants and the corresponding affinity constants. Importantly, a background response is generated due to the difference in the refractive indices of the running and sample buffers, as well as unspecific binding of the test compounds to the flow cell surface. This background is measured and subtracted by running the sample on a control flow cell coated with the same density of capture antibody in the absence of immobilized FAP. Furthermore, baseline drift correction of the binding data is performed, which is caused by slow dissociation of the captured FAP from the immobilized antibody. This drift is measured by injecting running buffer through a flow cell with the antibody and FAP immobilized to the sensor surface.

Biacore™ CM5 sensor chips were used. Human anti-FAP antibody (MAB3715, R&D systems) was diluted in 10 mM acetate buffer, pH 4.5, to a final concentration of 50 μg/mL. A 150 μL aliquot was transferred into plastic vials and placed into the sample rack of the Biacore™ T200 instrument. Amine Coupling Kit Reagent solutions were transferred into plastic vials and placed into 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 into plastic vials and placed into the sample rack. The Biacore™ liquid system was set-up as follows: Separate bottles containing distilled water (1 L), Running Buffer (500 mL), as well as an empty bottle for waste were placed onto the buffer tray. A preinstalled program for immobilization was used, with an immobilization level of 7000 RU. Immobilization was performed at 25° C. The immobilization procedure of anti-FAP antibodies was performed, as described in the Table 10.

TABLE 10

Immobilization protocol for anti-FAP antibodies used on the CM5 sensor chip.

Step
Injected solution
Contact time
Flow rate

Surface conditioning
50 mM NaOH
300 s
10 μL/min

Surface activation
EDC/NHS
420 s
10 μL/min

Washing
Ethanolamine
90 s
10 μL/min

Ligand binding
Human/mouse antibodies diluted
420 s
10 μL/min

in acetate buffer

Washing
Running Buffer
40 s
10 μL/min

Deactivation of reactive, non-
1M ethanolamine
420 s
10 μL/min

ligand bound surface

Washing
Running Buffer
30 s
10 μL/min

Human recombinant FAP was diluted in Running Buffer to a final concentration of 20 μg/mL. A 100 μL aliquot of human FAP-Working-Solution was transferred into plastic vials and placed into a sample rack. A 0.5 mM Compound-Stock-Solution was prepared by dissolving each compound in DMSO. For each test compound, Compound-Stock-Solutions were diluted in Running Buffer (HBST) at 500 nM and further diluted with HBST-DMSO Buffer (0.1% DMSO). SPR binding analyses for binary complexes were performed in SCK mode at 25° C. Table 11 describes the protocol for capturing and assessment of the binding kinetics. Following three SCK measurements, a baseline drift was assessed by injecting running buffer through a flow cell, with the antibody and FAP immobilized to the sensor surface.

TABLE 11

Protocol for assessing the binding kinetics.

Step
Injected solution
Contact time
Flow rate

Startup cycle as a triple run:
HBST-DMSO Buffer
60 s
30 μL/min

Washing & surface regeneration
10 mM glycine, pH 2
5 s

Binding target protein FAP
20 μg/mL rhFAP or
600 s
5 μL/min

(capturing)
4 μg/mL rmFAP

Washing (removal of unbound FAP)
HBST-DMSO-Buffer
2700 s
30 μL/min

1. Binding kinetics of test compound
Dilution no. 5 (0.19 nM)
120 s
30 μL/min

2. Binding kinetics of test compound
Dilution no. 4 (0.78 nM)
120 s
30 μL/min

3. Binding kinetics of test compound
Dilution no. 3 (3.125 nM)
120 s
30 μL/min

4. Binding kinetics of test compound
Dilution no. 2 (12.5 nM)
120 s
30 μL/min

5. Binding kinetics of test compound
Dilution no. 1 (50 nM)
120 s
30 μL/min

Dissociation cycle
HBST-DMSO Buffer
1800 s
30 μL/min

Regeneration
10 mM glycine, pH 2
7 s
30 μL/min

For each test compound, SPR raw data in the form of resonance units (RU) were plotted as sensorgrams using the Biacore™ T200 control software. The signal from the blank sensorgram was subtracted from that of the test compound sensorgram (blank corrected). The blank corrected sensorgram was corrected for baseline drift by subtracting the sensorgram of a SCK run without the test compound (running buffer only). The association rate (k_on), dissociation rate (k_off), dissociation constant (K_D), and t_1/2were calculated from Blank-normalized SPR data using the 1:1 Langmuir binding model from the Biacore™ T200 evaluation software. Raw data and fit results were imported as text files in IDBS. The pK_Dvalue (negative decadic logarithm of dissociation constant) was calculated in the IDBS excel template.

The results of this assay for a selection of compounds according to the present invention are presented in Table 12. Category A stands for pK_Dvalues >8.0, category B for pK_Dvalues between 7.1 and 8.0, category C for pK_Dvalues between 6.1 and 7.0.

TABLE 12

Compound ID, sequence and pkD category of Biacore assay

pK_D

ID
Sequence
Category

3BP-2974
Hex--C([3MeBn)-PPTEFC]DHFRD-NH2
A

3BP-2975
Ac-M-C([3MeBn)-PPTEFC]DHFRD-NH2
A

3BP-3105
Hex--C([3MeBn)-PPTEFC]DHFRD-Ttds--K(DOTA)--NH2
A

3BP-3168
DOTA--Ttds--Nle--C([3MeBn)-PPTEFC]DHFRD-NH2
A

3BP-3202
Ac--C([3MeBn)-PPTEFC]DHFRD-NH2
A

3BP-3275
Hex--C([3MeBn)-PPTEFC]DH-Nmf-R-Ttds--K(DOTA)--NH2
A

3BP-3288
Hex--C([3MeBn)-PPTEFC]D-NH2
A

3BP-3300
Hex--C([3MeBn)-PPTEFC]D-Gab-R-NH2
A

3BP-3301
Hex--C([3MeBn)-PPTEFC]D-Pamb-R-NH2
A

3BP-3319
DOTA--Ttds--Nle--C([3MeBn)-PPTEFC]-NH2
B

3BP-3320
DOTA--Ttds--Nle--C([3MeBn)-PPTEFC]D-NH2
A

3BP-3321
DOTA--Ttds--Nle--C([3MeBn)-PPTEFC]D-Pamb-R-NH2
A

3BP-3324
Hex--C([3MeBn)-PPTEFC]D-Mamb-R-NH2
A

3BP-3349
Hex--C([3MeBn)-PPTQFC]-NH2
A

3BP-3395
Hex--C([3MeBn)-PPTQFC]D-Ttds--K(DOTA)--NH2
A

3BP-3396
Hex--C([3MeBn)-PPTEFC]D-Ttds--K(DOTA)--NH2
A

3BP-3397
Hex--C([3MeBn)-PPTQFC]-Bhk(DOTA)--OH
A

3BP-3398
DOTA--Ttds--Nle--C([3MeBn)-PPTQFC]-Bal--OH
A

3BP-3401
DOTA--Ttds--Nle--C([3MeBn)-PPTQFC]D-NH2
A

3BP-3403
Hex--C([3MeBn)-PPTQFC]D-Ape--NH--DOTA
A

3BP-3407
Hex--[C(tMeBn(DOTA--PP))-PPTQFC]D-NH2
A

3BP-3426
Hex--C([tMeBn(DOTA--AET))-PPTQFC]D-NH2
A

3BP-3476
Hex--C([3MeBn)-PPTQFC]D-NH2
A

3BP-3489
Hex--C([3MeBn)-PPTQFC]-Bhk(DOTA--Ttds)--OH
A

3BP-3514
Pentyl--SO2--C([3MeBn)-PPTQFC]D-NH2
A

3BP-3554
Hex--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3555
Hex--C([tMeBn(DOTA--PP))-PPTQFC]-OH
A

3BP-3590
Hex--C([tMeBn(InDOTA--PP))-PPTQFC]D-NH2
A

3BP-3591
Hex--C([tMeBn(LuDOTA--PP))-PPTQFC]D-NH2
A

3BP-3592
Hex--C([tMeBn(GaDOTA--PP))-PPTQFC]D-NH2
A

3BP-3623
Hex--C([tMeBn(InDOTA--AET))-PPTQFC]-OH
A

3BP-3624
Hex--C([tMeBn(LuDOTA--AET))-PPTQFC]-OH
A

3BP-3650
Hex--C([tMeBn(DOTA--AET))-PPTQ-1Ni-C]-OH
A

3BP-3651
Hex--C([tMeBn(DOTA--AET))-PPTQFC]-Bal--OH
A

3BP-3652
Hex--C([tMeBn(DOTA--AET))-PPTQFC]-NH2
A

3BP-3653
Hex--C([tMeBn(DOTA--AET))-PPTEFC]-NH2
A

3BP-3654
Hex--C([tMeBn(DOTA--AET))-PPTQF-AET]
A

3BP-3656
Hex--C([tMeBn(DOTA--AET))-PPTQFC]G-OH
A

3BP-3657
Hex--C([tMeBn(DOTA--AET))-PPTQFC]-Gab--OH
A

3BP-3658
Hex--C([tMeBn(DOTA--AET))-PPTQFC]S-OH
A

3BP-3659
Hex--C([tMeBn(DOTA--AET))-PPTQFC]-Nmg--OH
A

3BP-3660
Hex--C([tMeBn(DOTA--AET))-PPTQFC]-Bhf--OH
A

3BP-3665
Hex--C([tMeBn(DOTA--AET))-PPTQFC]D-OH
A

3BP-3678
Hex--[C(tMeBn(DOTA--AET))--Nmg-PTQFC]-OH
A

3BP-3679
Hex--[C(tMeBn(DOTA--AET))-P-Hyp-TQFC]-OH
A

3BP-3680
Hex--[C(tMeBn(DOTA--AET))-PPTQ-Otf-C]-OH
A

3BP-3681
Hex--[C(tMeBn(DOTA--AET))-PPTQFC]d-NH2
A

3BP-3690
Pentyl--SO2--C([tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3692
Pentyl--SO2--C([tMeBn(DOTA--PP))-PPTQFC]D-NH2
A

3BP-3712
Hex--[C(tMeBn(InDOTA--AET))-PPTQFC]-NH2
A

3BP-3713
Hex--[C(tMeBn(InDOTA--AET))-PPTQF-AET]
A

3BP-3714
Hex--[C(tMeBn(InDOTA--AET))-PPTQFC]G-OH
A

3BP-3715
Hex--[C(tMeBn(InDOTA--AET))-PPTQFC]-Nmg--OH
A

3BP-3716
Hex--[C(tMeBn(InDOTA--AET))--Nmg-PTQFC]-OH
A

3BP-3717
Pentyl--SO2--[C(tMeBn(InDOTA--PP))-PPTQFC]D-NH2
A

3BP-3736
Hex--[C(tMeBn(DOTA--AET))-PPTQFC]-Bal--NH2
A

3BP-3737
Hex--[C(tMeBn(DOTA--AET))-PPTQFC]-Nmg--NH2
A

3BP-3907
iHex--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3910
Pent--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3918
EtOPr--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3940
nBu--CAyl--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3949
Hex--[C(tMeBn(GaDOTA--AET))-PPTQFC]-OH
A

3BP-4063
Hex--[C(tMeBn(N4Ac--AET))-PPTQFC]-OH
A

3BP-4064
Hex--[C(tMeBn(Cy5SO3--O2Oc--AET))-PPTQFC]-OH
A

3BP-4088
Hex--[C(tMeBn(N4Ac--O2Oc--AET))-PPTQFC]-OH
A

3BP-4147
Hex--[C(tMeBn(ReON4Ac--O2Oc--AET))-PPTQFC]-OH
A

3BP-4148
Hex--[C(tMeBn(ReON4Ac--AET))-PPTQFC]-OH
A

3BP-4161
N4Ac--Ttds--Nle--[C(3MeBn)-PPTEFC]-OH
A

3BP-4162
Hex--[C(tMeBn(NODAGA--AET))-PPTQFC]-OH
A

3BP-4168
Hex--[C(tMeBn(N4Ac--PP))-PPTQFC]-OH
A

3BP-4182
Hex--[C(tMeBn(ATTO488--AET))-PPTQFC]-OH
B

3BP-4184
Hex--[C(tMeBn(GaNODAGA--AET))-PPTQFC]-OH
A

3BP-4219
N4Ac--Ttds--Nle--[C(3MeBn)-PPTQFC]-OH
A

3BP-4221
N4Ac--PEG6--Nle--[C(3MeBn)-PPTEFC]-OH
A

3BP-4222
N4Ac-E-Ttds--Nle--[C(3MeBn)-PPTEFC]-OH
A

3BP-4232
Hex--[C(tMeBn(AF488--Ttds--Ttds--Ttds--Ttds--AET))-PPTQFC]-OH
C

3BP-4246
N4Ac--E(AGLU)--Ttds--Nle--[C(3MeBn)-PPTQFC]-OH
A

3BP-4249
N4Ac--E(AGLU)--Ttds--Nle--[C(3MeBn)-PPTEFC]-OH
A

3BP-4250
Hex--[C(tMeBn(DOTA--AET))-PPTEFC]-OH
A

3BP-4251
Hex--[C(tMeBn(NODAGA--O2Oc--AET))-PPTQFC]-OH
A

Example 33: PREP and DPP4 Protease Activity Assay

In order to test selectivity of FAP binding peptides toward both PREP and DPP4, protease activity assays were performed analogues to the FAP activity assay described above with following exceptions.

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

Fluorescence of AMC (excitation at 380 nm and emission at 460 nm) after cleavage was measured in a kinetic mode for 5 minutes at 37° C. in a SPECTRAmax M5 plate reader. RFU/sec was calculated by SoftMax Pro software and plotted against peptide concentration. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. The results of this assay for some of the compounds according to the present invention are given in the following Table 13.

TABLE 13

Results (pIC50 values) of PREP

and DPP4 activity assays

pIC50
pIC50

ID
(PREP)
(DPP4)

3BP-2881
<6
<6

3BP-3105
<6
<6

3BP-3168
<6
<6

3BP-3275
<6
<6

3BP-3287
<6
<6

3BP-3319
6.2
<6

3BP-3320
<6
<6

3BP-3321
<6
<6

3BP-3349
<6
<6

3BP-3397
<6
<6

3BP-3398
<6
<6

3BP-3407
<6
<6

3BP-3419
<6
<6

3BP-3426
<6
<6

3BP-3476
<6
<6

3BP-3554
<6
<6

Example 34: Specificity Screen

The specificity screening was carried out in order to early identify significant off-target interactions of compounds of the present invention. The specificity was tested using a standard battery of assays (“SafetyScreen44™ Panel”) comprising 44 selected targets and compounds binding thereto (referred to as “reference compounds”, Ref. Compounds), recommended by Bowes et al. (Bowes, et al., Nat Rev Drug Discov, 2012, 11: 909). The reference compounds served as positive controls for the respective assays, therefore inhibition is expected to be detected with these reference compounds. The compounds of the invention, however, were not expected to show inhibition in these assays. These binding and enzyme inhibition assays were performed by Eurofins Cerep SA (Celle l'Evescault, France).

3BP-3407 and 3BP-3554 were tested at 10 μM. Compound binding was calculated as % inhibition of the binding of a radioactively labeled ligand specific for each target (“% Inhibition of Specific Binding” (3BP-3407) or (3BP-3554), respectively). Compound enzyme inhibition effect was calculated as % inhibition of control enzyme activity.

Results showing an inhibition or stimulation higher than 50% are considered to represent significant effects of the test compounds. Such effects were not observed at any of the receptors studied which are listed in the following Table 14. The results of this assay are summarized in the following Table 14.

TABLE 14

Results of the specificity screening (SafetyScreen44 ™ Panel) for 10 μM 3BP-3407

and 10 μM 3BP-3554

% Inhibition of

Specific Binding

(3BP-
(3BP-
Ref

Cerep

Assay
3407)
3554)
Compound
Ki Ref [M]
Catalog Ref
Literature Reference

A2A (h) (agonist
−4
−16
NECA
2.90E−08
4
(Luthin, et al., Mol

radioligand)

Pharmacol, 1995, 47:

307)

alpha 1A (h)
2
−12
WB 4101
2.40E−10
2338
(Schwinn, et al., JBiol

(antagonist

Chem, 1990, 265: 8183)

radioligand)

alpha 2A (h)
−9
2
yohimbine
2.40E−09
13
(Langin, et al., EurJ

(antagonist

Pharmacol, 1989, 167:

radioligand)

95)

beta 1 (h) (agonist
4
−13
atenolol
3.40E−07
18
(Levin, et al., JBiol

radioligand)

Chem, 2002, 277:

30429)

beta 2 (h)
4
8
ICI 118551
1.60E−10
20
(Joseph, et al., Naunyn

(antagonist

Schmiedebergs
Arch

radioligand)

Pharmacol, 2004, 369:

525)

BZD (central)
−9
5
diazepam
8.10E−09
28
(Speth, et al., LifeSci,

(agonist

1979, 24: 351)

radioligand)

CB1 (h) (agonist
5
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CP 55940
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36
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radioligand)

J
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CB2 (h) (agonist
2
−5
WIN 55212-2
1.60E−09
37
(Munro, et al., Nature,

radioligand)

1993, 365: 61)

CCK1 (CCKA) (h)
24
16
CCK-8s
4.90E−11
39
(Bignon, et al., J

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D1 (h) (antagonist
0
7
SCH 23390
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44
(Zhou, et al., Nature,

radioligand)

1990, 347: 76)

D2S (h) (agonist
15
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7-OH-DPAT
1.30E−09
1322
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Acad
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USA, 1989,

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ETA (h) (agonist
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6
endothelin-1
1.50E−11
54
(Buchan, et al., BrJ

radioligand)

Pharmacol, 1994, 112:

1251)

NMDA (antagonist
9
1
CGS 19755
1.40E−07
66
(Sills, et al., EurJ

radioligand)

Pharmacol, 1991, 192:

19)

H1 (h) (antagonist
11
4
pyrilamine
1.10−09
870
(Smit, et al., BrJ

radioligand)

Pharmacol, 1996, 117:

1071)

H2 (h) (antagonist
−5
−16
cimetidine
4.30E−07
1208
(Leurs, et al., BrJ

radioligand)

Pharmacol, 1994, 112:

847)

MAO-A (antagonist
−5
−25
clorgyline
7.30E−10
443
(Cesura, et al., Mol

radioligand)

Pharmacol, 1990, 37:

358)

M1 (h) (antagonist
6
8
pirenzepine
2.90E−08
91
(Dorje, et al., J

radioligand)

Pharmacol
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Ther,

1991, 256: 727)

M2 (h) (antagonist
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7
Methoc-
4.80E−08
93
(Dorje, et al., J

radioligand)

tramine

Pharmacol
Exp
Ther,

1991, 256: 727)

M3 (h) (antagonist
10
1
4-DAMP
8.00E−10
95
(Peralta, et al., EmboJ,

radioligand)

1987, 6: 3923)

N neuronal alpha
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1.20E−09
3029
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Pharmacol
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(agonist

1996, 276: 289)

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delta (DOP) (h)
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1.20E−09
114
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(agonist

Pharmacol, 1994, 46:

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kappa (h) (KOP)
7
10
U50488
4.50E−10
4461
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(agonist

Natl
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mu (MOP) (h)
2
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DAMGO
3.70E−10
118
(Wang, et al., FEBSLett,

(agonist

1994, 338: 217)

radioligand)

5-HT1A (h)
−3
−5
8-OH-DPAT
2.20E−10
131
(Mulheron, et al., JBiol

(agonist

Chem, 1994, 269:

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5-HT1B(h)
−11
8
Serotonine
6.60E−08
4376
(Maier, et al., J

(antagonist

Pharmacol
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radioligand)

2009, 330: 342)

5-HT2A (h)
−2
4
(±)DOI
2.10E−10
471
(Bryant, et al., LifeSci,

(agonist

1996, 59: 1259)

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5-HT2B (h)
2
3
(±)DOI
4.20E−09
1333
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(agonist

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radioligand)

5-HT3 (h)
2
4
MDL 72222
6.50E−09
411
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(antagonist

Pharmacol, 1996, 118:

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GR (h) (agonist
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Dexame-
1.90E−09
469
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thasone

Ophthalmol
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AR (h) (agonist
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Testo-
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933
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sterone

Endocrinology, 1979,

104: 1007)

V1a (h) (agonist
16
1
[d(CH2)51,
1.WE−09
159
(Tahara, et al., BrJ

radioligand)

Tyr(Me)2]-

Pharmacol, 1998, 125:

AVP

1463)

Ca2+ channel (L,
42
54
nitrendipine
1.40E−10
161
(Gould, et al., ProcNatl

dihydropyridine

Acad
Sci
USA, 1982,

site) (antagonist

79: 3656)

radioligand)

Potassium Channel
2
6
Terfenadine
4.40E−08
4094
(Huang, et al., Assay

hERG (human)-

Drug
Dev
Technol, 2010,

[3H] Dofetilide

8: 727)

KV channel
−5
4
alpha-
9.70E−11
166
(Sorensen, et al., Mol

(antagonist

dendrotoxin

Pharmacol, 1989, 36:

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689)

Na+ channel (site
−7
14
veratridine
1.20E−05
169
(Brown, JNeurosci,

2) (antagonist

1986, 6: 2064)

radioligand)

norepinephrine
−8
−5
protriptyline
2.30E−09
355
(Pacholczyk, et al.,

transporter (h)

Nature, 1991, 350: 350)

(antagonist

radioligand)

dopamine
12
7
BTCP
6.80E−09
52
(Pristupa, et al., Mol

transporter (h)

Pharmacol, 1994, 45:

(antagonist

125)

radioligand)

5-HT transporter
−3
−8
imipramine
1.40E−09
439
(Tatsumi, et al., EurJ

(h) (antagonist

Pharmacol, 1999, 368:

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277)

COX1(h)
10
8
Diclofenac
1.30E−08
4173
(Vanachayangkul, et al.,

Enzyme
Res, 2012,

2012: 416062)

COX2(h)
−14
−22
NS398
5.40E−08
4186
(Vanachayangkul, et al.,

Enzyme
Res, 2012,

2012: 416062)

PDE3A (h)
−3
−37
milrinone
1.00E−06
4072
(Maurice, et al., NatRev

Drug
Discov, 2014, 13:

290)

PDE4D2 (h)
−5
−4
Ro 20-1724
2.30E−07
4077
(Maurice, et al., NatRev

Drug
Discov, 2014, 13:

290)

Lek kinase (h)
10
−4
Stauro-
2.30E−08
2906
(Park, et al., Anal

sporine

Biochem, 1999, 269: 94)

Acetylcholin-
−6
1
Galantha-
7.00E−07
363
(Ellman, et al., Biochem

esterase (h)

mine

Pharmacol, 1961, 7: 88)

Additionally, a specificity screen for proteases was performed by BPS Biosciences to further determine the specificity of the compounds of the invention (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).

3BP-3407 and 3BP-3554 were tested at 1 μM and 10 μM in duplicates. In the absence of the compound, the fluorescent intensity (Ft) in each data set was defined as 100% activity. In the absence of the enzyme, the background fluorescent intensity (Fb) in each data set was defined as 0% activity. The percent activity in the presence of each compound was calculated according to the following equation: % activity=(F−Fb)/(Ft−Fb), where F=the fluorescent intensity in the presence of the compound. Percentage inhibition was calculated according to the following formula: % inhibition=100%−% activity. Results showing an inhibition higher than 50% are considered to represent significant effects of the tested compound. The results of this assay are given in the following Table 15.

TABLE 15

Results of the specificity protease screening for 1 μM and

10 μM 3BP-3407 and 1 μM and 10 μM 3BP-3554

Percentage inhibition (%)

3BP-3407
3BP-3554

Enzyme
1 μM
10 μM
1 μM
10 μM
Reference

Activated Protein C
5
8
−11
1
74

(20 μM Dabigatran)

Beta secretase
−8
−5
1
7
84

(150 nM Verubecestat)

Caspase-3
1
−2
−2
−1
89

(100 nM Caspase 3/7 Inhibitor I)

Caspase-6
1
−1
6
−3
94

(1 μM Caspase 8 Inhibitor I)

Caspase-7
−3
−3
−1
−7
92

(1 μM Caspase 3/7 Inhibitor I)

Caspase-8
0
0
0
−3
87

(100 nM Caspase 8 Inhibitor 1)

Caspase-9
5
8
−1
−2
N/A

Cathepsin B
26
36
1
2
97

(100 nM E-64)

Cathepsin F
−3
−24
−23
−25
74

(1 μM Cystatin C)

Cathepsin L
3
6
0
−6
97

(1 μM E-64)

Cathepsin S
3
18
−10
−23
91

(100 nM E-64)

Cathepsin V
1
−18
−1
−1
83

(100 nM E-64)

A20
2
−4
1
0
99

(1 μM Ub-Aldehyde)

Ataxin3
1
10
2
−1
77

(10 μM Ub-Aldehyde)

Deubiquitinase
2
15
0
0
97

OTUD6B

(1 μM Ub-Aldehyde)

Ubiquitin carboxy-
−2
4
−4
4
92

terminal hydrolase L1

(100 nM Ub-Aldehyde)

Ubiquitin carboxy-
−1
14
0
0
95

terminal hydrolase L3

(10 nM Ub-Aldehyde)

Ubiquitin carboxyl-
3
7
0
−1
91

terminal hydrolase 2

(1 μM Ub-Aldehyde)

Ubiquitin carboxyl-
3
46
−4
−2
84

terminal hydrolase 5

(1 μM Ub-Aldehyde)

Ubiquitin carboxyl-
5
5
1
1
95

terminal hydrolase 7

(1 μM Ub-Aldehyde)

Ubiquitin carboxyl-
−3
6
2
1
73

terminal hydrolase 8

(1 μM Ub-Aldehyde)

Ubiquitin carboxyl-
−2
5
1
−1
82

terminal hydrolase 10

(1 μM Ub-Aldehyde)

Ubiquitin carboxyl-
−1
5
1
2
96

terminal hydrolase 14

(100 nM Ub-Aldehyde)

DPP3
ND
ND
2
−1
(100 nM Spinorphin)

DPP7
2
−3
−1
−7
83

(200 μM KR62436)

DPP8
1
5
1
11
96

(200 μM KR62436)

DPP9
−1
0
−1
−5
99

(200 μM KR62436)

FAP
98
99
97
99
100

(100 nM SP-13786)

serine protease
1
−68
−39
−372
94

NS3 (a.a. 3-181)

(100 nM Denoprevir)

from Hepatitis C

virus genotype 1a

(mutant D168V)

serine protease
1
5
−5
−9
100

NS3 (a.a. 3-181)

(100 nM Denoprevir)

from Hepatitis C

virus genotype 1b

serine protease
1
−6
−2
−17
99

NS3 (a.a. 3-181)

(100 nM Denoprevir)

from Hepatitis C

virus genotype 1b

(mutant D168V)

serine protease
−2
5
−1
0
90

NS3 (a.a. 3-181)

(100 nM Denoprevir)

from Hepatitis C

virus genotype 1b

(mutant R155K)

serine protease
0
2
0
−5
99

NS3 (a.a. 3-181)

(1 μM Denoprevir)

from Hepatitis C

virus genotype 1b

(mutant R155Q)

serine protease
0
−2
−13
−40
98

NS3 (a.a. 3-181)

(100 nM Denoprevir)

from Hepatitis C

virus genotype 2a

Matrix
−1
2
1
−7
87

metalloproteasel

(1 μM NNGH)

Matrix
3
3
−1
−2
95

metalloprotease 2

(100 nM NNGH)

Matrix
3
2
3
2
92

metalloprotease 9

(100 nM NNGH)

(mutant Q279R)

Renin
−1
3
0
−1
99

(30 nM Aliskiren)

Example 35: ¹¹¹In- and ¹⁷⁷Lu-Labeling of Selected Compounds

In order to serve as a diagnostically, therapeutically, or theragnostically active agent, a compound needs to be labeled with a radioactive isotope. The labeling procedure needs to be appropriate to ensure a high radiochemical yield and purity of the radiolabeled compound of the invention. This example shows that the compounds of the present invention are appropriate for radiolabeling and can be labeled in high radiochemical yield and purity.

30-100 MBq of ¹¹¹InCl₃(in 0.02 M HCl) were 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 down, DTPA and TWEEN-20 were added at a final concentration of 0.2 mM and 0.1%, respectively.

0.2-2.0 GBq ¹⁷⁷LuCl₃(in 0.04 M HCl) were 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) at a final buffer concentration of ˜0.4 M. The mixture was heated to 90° C. for 20 min. After cooling down, DTPA and TWEEN-20 were added at a final concentration of 0.2 mM and 0.1%, respectively.

In order to assess the long-term stability of ¹⁷⁷Lu-labeled compound in a formulation suitable for human use, after cooling down the reaction mixture was diluted with 9 volumes of a formulation buffer containing suitable stabilizers (e.g., ascorbate, methionine) and radiochemical purity was monitored over time.

The labeling efficiency was analyzed by thin layer chromatography (TLC) and HPLC. For TLC analysis, 1-2 μl of diluted labeling solution was applied to a strip of iTLC-SG chromatography paper (Agilent, 7.6×2.3 mm) and developed in citrate-dextrose solution (Sigma). The iTLC strip was then cut into 3 pieces and associated radioactivity was measured with a gamma-counter. The radioactivity measured at the solvent front represents free radionuclide and colloids, whereas the radioactivity at the origin represents radiolabeled compound. For HPLC, 5 μl of diluted labeling solution was analyzed with a Poroshell SB-C18 2.7 μm (Agilent). Eluent A: MeCN, eluent B: H₂O, 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 with the dead volume represents free radionuclide, the peak eluting with the peptide-specific retention time as determined with an unlabeled sample represents radiolabeled compound.

Radionuclidic incorporation yield was ≥90% and radiochemical purity ≥76% at end of synthesis. Exemplary radiochemical purities for ¹¹¹In-labeled compounds are shown in Table 16. ¹⁷⁷Lu-labeled compounds in formulations suitable for human use maintained a radiochemical purity of ≥90% up 6 days post synthesis (Table 17). The radiochromatograms for selected compounds are shown in FIGS. 1 to 4, whereby FIG. 1 shows a radiochromatogramm of ¹⁷⁷Lu-3BP-3407 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed immediately after synthesis, FIG. 2 shows a radiochromatogramm of ¹⁷⁷Lu-3BP-3407 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed six days after synthesis, FIG. 3 shows a radiochromatogramm of ¹⁷⁷Lu-3BP-3554 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed immediately after synthesis, and FIG. 4 shows a radiochromatogram of ¹⁷⁷Lu-3BP-3554 in formulation buffer containing 100 mg/mL ascorbate and 5 mg/mL L-methionine analyzed six days after synthesis.

TABLE 16

Radiochemical purity by HPLC of ¹¹¹In-labeled compounds.

HPLC
HPLC Area
HPLC Area %

retention
% at end
appr. 4 h post

time [min]
of synthesis
end of synthesis

¹¹¹In-3BP-3105
8.9
92.9
86.0

¹¹¹In-3BP-3168
7.9
94.2
92.3

¹¹¹In-3BP-3275
8.6
91.5
91.2

¹¹¹In-3BP-3320
7.4
97.7
96.5

¹¹¹In-3BP-3321
7.3
97.6
96.7

¹¹¹In-3BP-3397
8.3
76.3
77.6

¹¹¹In-3BP-3398
7.3
88.6
89.2

¹¹¹In-3BP-3407
7.3
97.6
95.4

¹¹¹In-3BP-3554
7.5
95.6
96.2

¹¹¹In-3BP-3652
7.3
87.1
88.8

¹¹¹In-3BP-3654
7.8
88.2
86.4

¹¹¹In-3BP-3656
7.3
87.8
87.1

¹¹¹In-3BP-3659
7.3
94.5
95.6

¹¹¹In-3BP-3678
7.4
89.9
89.2

¹¹¹In-3BP-3692
7.8
93.0
93.3

¹¹¹In-3BP-3767
7.4
94.6
92.9

¹¹¹In-3BP-3940
6.9
95.7
95.2

(3 h)

TABLE 17

Radiochemical purity by HPLC of ¹⁷⁷Lu-labeled

compounds in a formulation buffer containing

100 mg/mL ascorbate and 5 mg/mL L-methionine

analyzed on day 0 and day 6 post end of synthesis.

HPLC
HPLC
HPLC

retention
Area %
Area %

time [min]
Day 0
Day 6

¹⁷⁷Lu-3BP-3407
7.5
95.7
94.0

¹⁷⁷Lu-3BP-3554
7.6
97.2
95.6

Example 36: Imaging and Biodistribution Studies

Radioactively labeled compounds can be detected by imaging methods such as SPECT and PET. Furthermore, the data acquired by such techniques can be confirmed by direct measurement of radioactivity contained in the individual organs prepared from an animal injected with a radioactively labeled compound of the invention. Thus, the biodistribution (the measurement of radioactivity in individual organs) of a radioactively labeled compound can be determined and analyzed. This example shows that the compounds of the present invention show a biodistribution appropriate for diagnostic imaging and therapeutic treatment of tumors.

All animal experiments were conducted in compliance with the German animal protection laws. Male SCID beige (6- to 8-week-old, Charles River, Sulzfeld, Germany) were inoculated with 5×10⁶HEK-FAP (embryonic human kidney 293 cells genetically engineered to express high levels of FAP) cells in one shoulder. When tumors reached a size of >150 mm³mice received ˜30 MBq ¹¹¹In-labelled compounds of the invention (diluted to 100 μL with PBS) administered intravenously via the tail vein. Images were obtained on a NanoSPECT/CT system (Mediso Medical Imaging Systems, Budapest, Hungary) using exemplarily the following acquisition and reconstruction parameters (Table 18).

TABLE 18

Acquisition and reconstruction parameters of NanoSPECT/CT imaging

Acquistion parameters SPECT

System
NanoSPECT/CT ™

Scan range
whole body, 3-bed holder (mouse hotel)

Time per projection
60 s

Aperture model,
Aperture #2, 1.5 mm

pinhole diameter

Reconstruction parameters

Method
HiSPECT (Scivis), iterative reconstruction

Smoothing
35%

Iterations
9

Voxel size
0.15 mm × 0.15 mm × 0.15 mm

Acquisition parameters CT

System
NanoSPECT/CT ™

Scan range
whole body, 3-bed holder (mouse hotel)

Scan duration
7 minutes

Tube voltage
45 kVp

Exposure time
500 ms

Number of projections
240

Imaging data were saved as DICOM files and analysed 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 at 24h or 48h post injection and then dissected. Different organs and tissues were collected and weighed, and the radioactivity was determined by γ-counting. Two animals were used per time point. Results are expressed as a percentage of injected dose per gram of tissue (% ID/g).

The results of the imaging and biodistribution studies for selected compounds are shown in FIGS. 5-14, 20 and 21.

Example 37: Efficacy Study—HEK-FAP

Radioactively labeled compounds can be used for therapeutic and diagnostic application in various diseases, especially cancer. This example shows that the compounds of the present invention have anti-tumor activity suitable for the therapeutic treatment of tumors.

All animal experiments were conducted in compliance with the German animal protection laws. Female swiss nude mice (7- to 8-week-old, Charles River Laboratories, France) were inoculated with 5×10⁶HEK-FAP cells in one shoulder, and treatments were administered when the tumors reached a mean tumor volume of of 160±44 mm³. Mice were divided into 4 different groups of 10 animals/group: Group 1—vehicle control, Group 2—cold compound ^natLu-3BP-3554, Group 3-30 MBq ¹⁷⁷Lu-3BP-3554 (low dose), and Group 4-60 MBq ¹⁷⁷Lu-FAP-3554 (high dose). Treatments were administered on Day 0 by intravenous injection into the tail vein at 4 mL/kg (100 L/mouse). Tumor volume and body weights were measured at Day 0 (i.e. the first day of radiotracer administration) and then thrice weekly until completion of the study.

The tracer distribution in mice injected with ¹⁷⁷Lu-labeled 3BP-3554 was determined by SPECT imaging in three mice dosing group. Subsequently, following SPECT, a CT scan was done for anatomical information. Imaging was performed 3 h, 24 h, 48 h and 120 h post injection with a NanoSPECT/CT system (Mediso Medical Imaging Systems, Budapest, Hungary) using exemplarily the following acquisition and reconstruction parameters (Table 19).

TABLE 19

Acquisition and reconstruction parameters of NanoSPECT/CT imaging

Acquistion parameters SPECT

System
NanoSPECT/CT ™

Scan range
whole body, 3-bed holder (mouse hotel)

Time per projection
60 s or 120 s

Aperture model,
Aperture #2, 1.5 mm

pinhole diameter

Reconstruction parameters

Method
HiSPECT (Scivis), iterative reconstruction

Smoothing
35%

Iterations
9

Voxel size
0.15 mm × 0.15 mm × 0.15 mm

Acquisition parameters CT

System
NanoSPECT/CT ™

Scan range
whole body, 3-bed holder (mouse hotel)

Scan duration
7 minutes

Tube voltage
45 kVp

Exposure time
500 ms

Number of projections
240

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

Tumors in vehicle and cold compound ^natLu-3BP-3554-treated mice reached a mean tumor volume (MTV) of 1338±670 mm³and 1392±420 mm³on day 14, respectively (FIG. 15 A). Statistically significant (P<0.01) anti-tumor activity was observed in mice of both treatment groups. Tumor growth inhibition (TGI) at day 14 was 111% and 113% in mice treated with a single dose of 30 or 60 MBq ¹⁷⁷Lu-3BP-3554, respectively, relative to the vehicle-treated group. The MTV in all mice treated with ¹⁷⁷Lu-3BP-3554 was reduced to ≤70 mm³on day 14. Tumors were monitored for regrowth on day 42 (which represents the end of the study) three of ten and nine of ten mice treated with 30 or 60 MBq ¹⁷⁷Lu-3BP-3554, respectively, were tumor-free (<10 mm³), suggesting a potential dose-response in this model. No treatment-related body weight loss was observed throughout the study (FIG. 15 B). After a 3-5% decrease in body weight observed in all groups on Day 2, the body weight of the animals increased over time.

SPECT/CT imaging of 3 animals of both ¹⁷⁷Lu-labeled treatment groups showed high tumor-to-background contrast during all examined time points (3-120 h post-injection (p.i.)). High tumor retention up to 120 h was observed. The organ with the highest non-target uptake was the kidney, with tumor-to-kidney ratios of 8.6±0.6 and 8.0±1.6 at 3 h p.i. in mice treated with 30 or 60 MBq ¹⁷⁷Lu-3BP3554, respectively. These ratios increased over time, attaining the highest value at 120 h with 40±7.9 and 32±7.4 tumor-to-kidney ratios in mice treated with 30 or 60 MBq ¹⁷⁷Lu-3BP3554, respectively. An exemplary panel of SPECT/CT images for mouse 5 which was a high dose animal is shown in FIG. 16 A and for mouse 1 which was a low-dose animal is shown in FIG. 16 B.

Example 38: Imaging Study—Sarcoma PDX Models

Sarcoma tumors have been reported to express FAP, and imaging of four different sarcoma patient-derived xenograft (PDX) tumor models was performed to evaluate 3BP-3554 uptake. The Sarc4183, Sarc4605, Sarc4809 and Sarc12616 PDX models were derived from patients with rhabdomyosarcoma, osteosarcoma, undifferentiated sarcoma and undifferentiated pleiomorphic sarcoma, respectively (Experimental Pharmacology & Oncology Berlin-Buch, Germany). Tumor fragments were transplanted subcutaneously in the left flank of 8-week-old NMRI nu/nu mice (Janvier Labs, France). All animal experiments were conducted in compliance with the German animal protection laws. 47 days (Sarc4183, Sarc4809) or 46 days (Sarc4605, Sarc12616) after transplantation, 2-3 mice per model were imaged 3 hours after a single intravenous injection of 30 MBq of ¹¹¹In-3BP-3554. Imaging was performed as described in Example 36.

The imaging results with ¹¹¹In-3BP-3554 showed high tumor uptake 3 h p.i. and a high tumor-to-background contrast. Representative SPECT/CT images are shown in FIG. 17 A. Quantification of tumor uptake of two (Sarc4605, Sarc12616) or three (Sarc4183, Sarc4809) PDX-bearing mice, respectively, revealed % 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) as shown in FIG. 17 B. These results demonstrate ¹¹¹In-3BP-3554 uptake in all 4 sarcoma models. 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).

Example 39: Efficacy Study—Sarcoma Sarc4809 PDX Model

The efficacy of ¹⁷⁷Lu-3BP-3554 was investigated in the human sarcoma PDX tumor model Sarc4809. This model of an undifferentiated sarcoma demonstrates ¹¹¹In-3BP-3554 uptake (Example 38) and was also shown to express FAP by immunohistochemistry.

All animal experiments were conducted in compliance with the German animal protection laws. Sarc4809 tumor fragments were transplanted subcutaneously at the left flank of 8-week-old NMRI nu/nu mice (Janvier Labs, France). Treatment started 23 days after transplantation at a mean tumor volume of 187.08±123.8 mm³. Mice were split into four groups of 10 animals/group: Group 1—vehicle control, Group 2—cold compound ^natLu-FAP-3554, Group 3—30 MBq ¹⁷⁷Lu-3BP-3554, Group 4-60 MBq ¹⁷⁷Lu-FAP-3554. Treatments were administered on Day 0 by intravenous injection into the tail vein at 4 mL/kg (100 L/mouse). Tumor volume and body weight were determined at Day 0 (i.e. the first day of radiotracer administration) and then thrice weekly until completion of the study.

All tumors continuously grew throughout the follow-up period of the study until day 42. Tumors in vehicle and ^natLu-3BP-3554 treated mice (control groups) reached an MTV of 894 610 mm³and 1225±775 mm³on day 31 (the last day on which at least 50% mice per group were still alive), respectively. Tumors in mice treated with a single dose of 30 or 60 MBq ¹⁷⁷Lu-3BP-3554 reached an MTV of 635±462 and 723±391 mm³on day 31, respectively (FIG. 18A). Statistically significant (P<0.05) anti-tumor activity was observed in mice of both treatment groups. Tumor growth inhibition (TGI) at day 31 was 61% and 73% in mice treated with a single dose of 30 or 60 MBq ¹⁷⁷Lu-3BP-3554, respectively, relative to the vehicle-treated group. No treatment-related body weight loss (BWL) was observed throughout the study. In all groups body weight increased during study follow-up (FIG. 18B).

Example 40: Pharmacokinetic Studies

The pharmacokinetic behavior of selected compounds was assessed in mice and rats. This characterization of the pharmacokinetic behavior of a compound enables new insights into distribution and elimination of the compound and the calculation of the exposure.

Different amounts of the compounds were stable formulated in PBS. The formulations were applied intravenous with a dose of 4 nmol/kg, 40 nmol/kg and 400 nmol/kg in mice and 2 nmol/kg, 20 nmol/kg and 200 nmol/kg (3BP-3554) or 40 nmol/kg and 400 mol/kg (3BP-3623) in rats. Assuming an allometric translation factor of 12.3 from human to mouse, and 6.2 from human 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.

Blood samples were collected after different times (5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h) from tail vein (rats) or retrobulbar (mice).

After separation of the blood cells from the blood plasma by centrifugation, the compounds were quantified in the prepared plasma samples were subjected to a protein precipitation procedure. 150 μl of a zinc sulphate precipitation agent containing 78% 0.1 M zinc sulphate and 22% acetonitrile was added. After incubation at room temperature for 30 min the precipitate was separated by centrifugation. To 100 μl of the supernatant 10 μl of 1% formic acid was added followed by further incubation at 60° C. for 10 min to complete the formation of the zinc chelate, if the compound contains a free DOTA moiety.

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

Mass spectrometric detection was performed in positive ion ESI mode by multiple reaction monitoring (MRM) with detection parameters as described in Table 20.

TABLE 20

Mass spectrometric detection parameters

Collision

Compound
Fragmentor

Precursor
Product
energy

3BP-4343
190 V
Quantifier
767.0
683.2
24 V

Qualifier
767.0
542.9
38 V

3BP-3623
110 V
Quantifier
791.8
777.6
21 V

Qualifier
791.8
708.2
19 V

Quantitation of test items was accomplished using the Quantitative Analysis software of the Agilent MassHunter software suite. A quadratic regression was performed with a weighting factor of 1/x.

The plasma level was subjected to a non-compartmental analysis (NCA) with following results: initial concentration of the compound (C₀), volume of distribution at steady state (V_ss), volume of distribution in the terminal phase (V_z), terminal half-life (t_1/2), clearance (CL) and area under the curve extrapolated to infinity (AUC_inf). A summary of NCA parameters of 3BP-3554 are presented in Table 21 for 3BP-3554 in mouse plasma and in Table 22 for 3BP-3554 in rat plasma, and of NCA parameters of 3BP-3623 in Table 23 for 3BP-3623 in mouse plasma and in Table 24 for 3BP-3623 in rat plasma.

TABLE 21

Summary of NCA parameters of 3BP-3554 in mouse plasma

PK parameter
4 nmol/kg
40 nmol/kg
400 nmol/kg

C₀
25.6
nM
177
nM
4970
nM

V_ss
0.21
L/kg
0.32
L/kg
0.10
L/kg

V_z
0.26
L/kg
1.02
L/kg
0.21
L/kg

AUC_inf
8.3
nM h
56
nM h
961
nM h

t_1/2
23
min
59
min
40
min

CL
0.482
L/kg h
0.711
L/kg h
0.482
L/kg h

TABLE 22

Summary of NCA parameters of 3BP-3554 in rat plasma

PK parameter
2 nmol/kg
20 nmol/kg
200 nmol/kg

C₀
10.3
nM
111
nM
1480
nM

V_ss
0.28
L/kg
0.30
L/kg
0.17
L/kg

V_z
0.32
L/kg
0.35
L/kg
0.42
L/kg

AUC_inf
8.1
nM h
69
nM h
726
nM h

t_1/2
54
min
50
min
63
min

CL
0.248
L/kg h
0.291
L/kg h
0.275
L/kg h

TABLE 23

Summary of NCA parameters of 3BP-3623 in mouse plasma

PK parameter
4 nmol/kg
40 nmol/kg
400 nmol/kg

C₀
17.6
nM
228
nM
2134
nM

V_ss
0.36
L/kg
0.31
L/kg
0.20
L/kg

V_z
0.44
L/kg
0.53
L/kg
0.64
L/kg

AUC_inf
7.7
nM h
55
nM h
532
nM h

t_1/2
35
min
30
min
35
min

CL
0.518
L/kg h
0.722
L/kg h
0.752
L/kg h

TABLE 24

Summary of NCA parameters of 3BP-3623 in rat plasma

PK parameter
40 nmol/kg
400 nmol/kg

C₀
127
nM
1408
nM

V_ss
0.48
L/kg
0.32
L/kg

V_z
0.58
L/kg
0.93
L/kg

AUC_inf
74
nM h
738
nM h

t_1/2
45
min
71
min

CL
0.541
L/kg h
0.542
L/kg h

The results indicate distribution mainly in the blood and interstitial fluids and a clearance typical for peptides with terminal half-lifes between 23 min and 59 min in mice and between 45 min and 71 min in rats. Exposure as described by the AUC correlates almost linear to the injected dose and the clearance is constant for all applied doses in a particular animal model. These observations suggest no significant non-linearity of the pharmacokinetic behavior that need to be considered for first-in-human dose calculation.

The features of the present 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 various forms thereof.

Example 41: Synthesis of nBu-CAyl-[Cys(tMeBn(NOPO-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (3BP-4768)

Similar methods to the synthesis of 3BP-3940 (Example 8b, and Cyclization method B) were used for the synthesis of the title compounds. In contrast to 3BP-3940, the AET amine moiety was conjugated to a preactivated NOPO-chelator instead of DOTA-NHS. Briefly, to a solution of nBu-CAyl-[Cys(tMeBn(H-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (78.0 mg, 71.8 μmol) in 1 ml DMF, 22 μl of DIPEA were added to adjust the pH value to approximately 7.5-8. To a solution of NOPO (35.6 mg, 71.8 μmol, 1 eq.) and DIPEA (50 μl, 288 μmol, 4 eq.) in a mixture of DMF/DMSO (1:1, v/v, 500 μl) was added a solution of HATU (55 mg, 143.6 μmol, 2 eq.) in 250 μl DMF. After only a second of preactivation, the chelator mix was immediately transferred to the peptide, and the pH of the obtained solution was re-adjusted to 8 with DIPEA. For full conversion, the NOPO conjugation step was repeated once. After completion of the reaction as judged by LC/TOF-MS, volatiles were removed in vacuo followed by lyophilization from a mixture of water/acetonitrile. The crude product was subjected to HPLC purification (20 to 40% B in 15 min—Kinetex) to yield 35.95 mg of the pure title compound (32.0% yield). HPLC: R_t=6.0 min. LC/TOF-MS: exact mass 1561.572 (calculated 1561.574). C₆₄H₁₀2N₁₃O₂₀P₃S₃(MW=1562.692).

Example 42: FACS Binding Assay

In order to determine binding of compounds according to the present invention to FAP-expressing cells, a competitive FACS binding assay was established.

FAP-expressing human WI-38 fibroblasts (ECACC) were cultured in EMEM including 15% fetal bovine serum, 2 mM L-Glutamine and 1% Non-essential amino acids. Cells were detached with Accutase (Biolegend, #BLD-423201) and washed in FACS buffer (PBS including 1% FBS). Cells were diluted in FACS buffer to a final concentration of 100.000 cells per ml and 200 μl of the cell suspension are transferred to a u-shaped non-binding 96-well plate (Greiner). Cells were washed in 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) in the presence of increasing concentrations of peptides at 4° C. for 1 hour. Cell were washed twice with FACS buffer and resuspended in 200 μl FACS buffer. Cells were analyzed in an Attune NxT flow cytometer. Median fluorescence intensities (Cy5 channel) was calculated by Attune NxT software and plotted against peptide concentrations. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. The results of this assay as well as the ones of the FAP protease activity assay as subject to Example 43 for each compound according to the present invention are presented in Table 25 (shown in Example 43). pIC50 category A stands for pIC50 values >8.0, category B for pIC50 values between 7.1 and 8.0, category C for pIC50 values between 6.1 and 7.0 and category D for pIC50 values ≤6.0.

Example 43: FAP Protease Activity Assay

In order to determine the inhibitory activity of compounds according to the present invention to FAP-expressing cells, a FRET-based FAP protease activity assay was established.

Recombinant human FAP (R&D systems, #3715-SE) was diluted in assay buffer (50 mM Tris, 1 M NaCl, 1 mg/mL BSA, pH 7.5) to a concentration of 3.6 nM. 25 μl of the FAP solution was mixed with 25 μl of a 3-fold serial dilution of the test compounds and incubated for 5 min in a white 96-well ProxiPlate (Perkin Elmer). As specific FAP substrate the FRET-peptide HiLyteFluor™ 488—VS(D-)P SQG K(QXL® 520)—NH2 was used (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. prior to use. Substrate cleavage and increase in fluorescence (excitation at 485 nm and emission at 538 nm) was measured in a kinetic mode for 5 minutes at 37° C. in a SPECTRAmax M5 plate reader. RFU/sec was calculated by SoftMax Pro software and plotted against peptide concentration. Four parameter logistic (4PL) curve fitting and pIC50 calculations were performed using ActivityBase software. The results of this assay for each compound according to the present invention are given in Table 9 and Table 25. pIC50 category A stands for pIC50 values >8.0, category B for pIC50 values between 7.1 and 8.0, category C for pIC50 values between 6.1 and 7.0 and category D for pIC50 values ≤6.0.

As evident from Table 25, the compounds of the present invention show surprisingly superior results in both the FACS Binding assay and the FAP protease activity assay.

TABLE 25

Compound ID, sequence, exact calculated mass, exact mass found, retention time in

minutes as determined by HPLC and pIC50 category of FACS binding and FAP activity assay

Exact
Exact

pIC50
pIC50

Mass
Mass
R_t
Category
Category

ID
Sequence
(calc)
(found)
(HPLC)
(FACS)
(activity)

3BP-4384
Hex-[C(tMeBn(DATA-Ttds-AET))-PPTQFC]-
1770.83
1770.88
6.91
B
B

OH

3BP-4541
N4Ac-PPAc-Ttds-Nle-[C(3MeBn)-PPTQFC]-
1623.85
1623.85
5.56
B
B

OH

3BP-4549
N4Ac-Ttds-Nle-[C(3Lut)-PPTQFC]-OH
1498.77
1498.82
4.32
B
A

3BP-4550
N4Ac-PEG6-Nle-[C(3Lut)-PPTQFC]-OH
1531.78
1531.84
4.45
C
B

3BP-4551
N4Ac-PEG6-Nle-[C(3MeBn)-PPTQFC]-OH
1530.78
1530.85
5.72
B
B

3BP-4552
N4Ac-PEG6-Ttds-Nle-[C(3MeBn)-PPTQFC]-
1832.97
1833.04
5.78
B
B

OH

3BP-4634
Hex-[C(tMeBn(GaDATA-Ttds-AET))-
1836.73
1836.76
6.89
B
B

PPTQFC]-OH

3BP-4695
Hex-[C(tMeBn(NODAGA-AET))-PPTEFC]-OH
1441.60
1441.65
6.71
B
B

3BP-4708
Hex-[C(tMeBn(NODAGA-O2Oc-AET))-
1584.70
1584.76
6.62
A
A

PPTQFC]-NH2

3BP-4713
NODAGA-Ttds-Nle-[C(3MeBn)-PPTQFC]-OH
1668.78
1668.81
6.47
B
B

3BP-4714
NODAGA-Ttds-Nle-[C(3MeBn)-PPTEFC]-OH
1669.76
1669.81
6.50
B
B

3BP-4723
nBu-CAyl-[C(tMeBn(NODAGA-AET))-
1441.61
1441.66
6.13
A
A

PPTQFC]-OH

3BP-4724
nBu-CAyl-[C(tMeBn(NODAGA-AET))-
1442.59
1442.64
6.83
A
B

PPTEFC]-OH

3BP-4729
Hex-[C(tMeBn(NOPO-AET))-PPTQFC]-NH2
1559.59
1559.67
6.46
A
A

3BP-4743
NODAGA-Ttds-Nle-[C(3MeBn)-PPTEFC]-NH2
1668.78
1668.82
6.44
B
B

3BP-4744
nBu-CAyl-[C(tMeBn(InDOTA-AET))-
1579.53
1579.54
5.70
A
A

PPTQFC]-NH2

3BP-4745
nBu-CAyl-[C(tMeBn(LuDOTA-AET))-
1641.57
1641.61
5.91
A
A

PPTQFC]-NH2

3BP-4768
nBu--CAyl-[C(tMeBn(NOPO-AET))-PPTQFC]-
1561.57
1561.59
6.05
A
A

OH

3BP-4769
nBu-CAyl-[C(tMeBn(GaDOTA-AET))-
1536.54
1536.51
5.94
A
A

PPTQFC]-OH

3BP-4773
N4Ac-PEG6-Nle-[C(3Lut)-PPTEFC]-OH
1532.76
1532.85
4.36
B
B

3BP-4774
N4Ac-PPAc-Ttds-Nle-[C(3Lut)-PPTQFC]-OH
1624.85
1624.94
4.12
B
B

3BP-4775
N4Ac-PPAc-Ttds-Nle-[C(3Lut)-PPTEFC]-OH
1625.83
1625.94
4.45
B
B

3BP-4778
nBu-CAyl-[C(tMeBn(NOPO-AET))-PPTEFC]-
1562.56
1562.65
6.08
A
A

OH

3BP-4779
N4Ac-PEG6-Bal-Nle-[C(3MeBn)-PPTEFC]-
1602.80
1602.93
5.61
C
D

OH

3BP-4780
N4Ac-PEG6-Ttds-Nle-[C(3MeBn)-PPTEFC]-
1833.95
1834.09
5.73
B
A

OH

3BP-4781
N4Ac-PPAc-PEG6-Nle-[C(3MeBn)-PPTEFC]-
1657.85
1657.97
5.70
B
B

OH

3BP-4782
N4Ac-PPAc-Ttds-Nle-[C(3MeBn)-PPTEFC]-
1624.84
1624.96
5.54
B
B

OH

3BP-4783
N4Ac-PPAc-Bal--Nle-[C(3MeBn)-PPTEFC]-OH
1393.69
1393.79
5.35
D
D

3BP-4784
N4Ac-PPAc-Ttds-Nle-[C(3MeBn)-PPTEFC]-
1695.87
1696.00
5.55
A
A

Bal-OH

3BP-4785
N4Ac-PPAc-Ttds-Nle-[C(3MeBn)-PPTEFC]D-
1739.86
1739.99
5.52
A
A

OH

3BP-4816
Hex-[C(tMeBn(GaNOPO-AET))-PPTEFC]-OH
1627.46
1627.54
6.79
B
B

3BP-4818
Hex-[C(tMeBn(NOPO-AET))-PPTEFC]-OH
1561.56
1561.67
6.55
B
B

3BP-4844
nBu-CAyl-[C(tMeBn(LuDOTA-PP))-PPTQFC]-
1650.62
1650.70
5.58
A
A

NH2

3BP-4960
N4Ac-PPAc-Ttds-Nle-[C(3MeBn)-PPTQFC]-
1694.89
1695.06
5.20
A
A

Bal-OH

3BP-4961
N4Ac-PPAc-Ttds-Nle-[C(3Lut)-PPTQFC]-Bal-
1695.88
1696.05
4.06
A
A

OH

3BP-5201
NOTA-Ttds-Nle-[C(3MeBn)-PPTQFC]-OH
1596.76
1596.84
6.33
A
B

3BP-5210
nBu-CAyl-[C(tMeBn(NOTA-AET))-PPTQFC]-
1369.59
1369.58
6.16
A
A

OH

3BP-5260
nBu-CAyl-[C(tMeBn(GaNOPO-AET))-
1627.48
1627.48
6.15
A
A

PPTQFC]-OH

3BP-5261
GaNODAGA-Ttds-Nle-[C(3MeBn)-PPTQFC]-
1734.68
1734.68
6.55
A
A

OH

3BP-5262
GaNOTA-Ttds-Nle-[C(3MeBn)-PPTQFC]-OH
1663.67
1663.66
6.17
A
A

3BP-5263
nBu-CAyl-[C(tMeBn(GaNOTA-AET))-
1436.50
1436.49
5.70
A
A

PPTQFC]-OH

3BP-5264
Hex-[C(tMeBn(GaNOTA-AET))-PPTQFC]-OH
1435.50
1435.49
6.26
A
A

3BP-5273
Hex-[C(tMeBn(AcPCTA-AET))-PPTQFC]-OH
1519.62
1519.62
6.45
A
A

3BP-5288
Hex-[C(tMeBn(LSC-AET))-PPTQFC]-OH
1468.66
1468.65
6.13
A
A

3BP-5315
nBu-CAyl-[C(tMeBn(AlFNOTA-AET))-
1413.55
1413.55
6.26
text missing or illegible when filed

A

PPTQFC]-OH

3BP-5323
Hex-[C(tMeBn(DOTAM-AET))-PPTQFC]-OH
1466.69
1466.69
5.86
text missing or illegible when filed

A

3BP-5363
Hex-[C(tMeBn(LaAcPCTA-AET))-PPTQFC]-
1654.50
1654.50
6.75
text missing or illegible when filed

B

OH

N.D.—not determined

text missing or illegible when filed

indicates data missing or illegible when filed

Example 44: Surface Plasmon Resonance Assay

TABLE 26

Immobilization protocol for anti-FAP antibodies used on the CM5 sensor chip.

Step
Injected solution
Contact time
Flow rate

Surface conditioning
50 mM NaOH
300
s
10 μL/min

Surface activation
EDC/NHS
420
s
10 μL/min

Washing
Ethanolamine
90
s
10 μL/min

Ligand binding
Human/mouse
420
s
10 μL/min

antibodies diluted in

acetate buffer

Washing
Running Buffer
40
s
10 μL/min

Deactivation of reactive,
1M ethanolamine
420
s
10 μL/min

non-ligand bound surface

Washing
Running Buffer
30
s
10 μL/min

Human recombinant FAP was diluted in Running Buffer to a final concentration of 20 μg/mL. A 100 μL aliquot of human FAP-Working-Solution was transferred into plastic vials and placed into a sample rack. A 0.5 mM Compound-Stock-Solution was prepared by dissolving each compound in DMSO. For each test compound, Compound-Stock-Solutions were diluted in Running Buffer (HBST) at 500 nM and further diluted with HBST-DMSO Buffer (0.1% DMSO). SPR binding analyses for binary complexes were performed in SCK mode at 25° C. Table 27 describes the protocol for capturing and assessment of the binding kinetics. Following three SCK measurements, a baseline drift was assessed by injecting running buffer through a flow cell, with the antibody and FAP immobilized to the sensor surface.

TABLE 27

Protocol for assessing the binding kinetics.

Step
Injected solution
Contact time
Flow rate

Startup cycle as a triple run:
HBST-DMSO Buffer
60
s
30 μL/min

Washing & surface regeneration
10 mM glycine, pH 2
5
s

Binding target protein FAP
20 μg/mL rhFAP or
600
s
5 μL/min

(capturing)
4 μg/mL rmFAP

Washing (removal of unbound FAP)
HBST-DMSO-Buffer
2700
s
30 μL/min

1. Binding kinetics of test compound
Dilution no. 5 (0.19 nM)
120
s
30 μL/min

2. Binding kinetics of test compound
Dilution no. 4 (0.78 nM)
120
s
30 μL/min

3. Binding kinetics of test compound
Dilution no. 3 (3.125 nM)
120
s
30 μL/min

4. Binding kinetics of test compound
Dilution no. 2 (12.5 nM)
120
s
30 μL/min

5. Binding kinetics of test compound
Dilution no. 1 (50 nM)
120
s
30 μL/min

Dissociation cycle
HBST-DMSO Buffer
1800
s
30 μL/min

Regeneration
10 mM glycine, pH 2
7
s
30 μL/min

The results of this assay for a selection of compounds according to the present invention are presented in Table 28. Category A stands for pK_Dvalues >8.0, category B for pK_Dvalues between 7.1 and 8.0, category C for pK_Dvalues between 6.1 and 7.0.

TABLE 28

Compound ID, sequence and pkD category of Biacore assay

pK_D

ID
Sequence
Category

3BP-3907
iHex--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3910
Pent--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3918
EtOPr--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3940
nBu--CAyl--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-3941
nBu--COyl--[C(tMeBn(DOTA--AET))-PPTQFC]-OH
A

3BP-4425
nBu--CAyl--[C(tMeBn(LuDOTA--AET))-PPTQFC]-OH
A

3BP-4426
nBu--CAyl--[C(tMeBn(InDOTA--AET))-PPTQFC]-OH
A

3BP-4541
N4Ac--PPAc--Ttds--Nle--[C(3MeBn)-PPTQFC]-OH
B

3BP-4549
N4Ac--Ttds--Nle--[C(3Lut)-PPTQFC]-OH
B

3BP-4550
N4Ac--PEG6--Nle--[C(3Lut)-PPTQFC]-OH
B

3BP-4551
N4Ac--PEG6--Nle--[C(3MeBn)-PPTQFC]-OH
B

3BP-4560
nBu--CAyl--[C(tMeBn(DOTA--AET))-PPTQFC]-NH2
A

3BP-4564
nBu--CAyl--[C(tMeBn(DOTA--PP))-PPTQFC]-OH
A

3BP-4565
nBu--CAyl--[C(tMeBn(DOTA--PP))-PPTQFC]-NH2
A

3BP-4607
nBu--CAyl--[C(tMeBn(DOTA--AET))-Nmg-PTQFC]-OH
A

3BP-4621
nBu--CAyl--[C(tMeBn(DOTA--AET))-Nmg-PTQFC]-NH2
A

3BP-4744
nBu--CAyl--[C(tMeBn(InDOTA--AET))-PPTQFC]-NH2
A

3BP-4745
nBu--CAyl--[C(tMeBn(LuDOTA--AET))-PPTQFC]-NH2
A

3BP-4768
nBu--CAyl--[C(tMeBn(NOPO--AET))-PPTQFC]-OH
A

3BP-4769
nBu--CAyl--[C(tMeBn(GaDOTA--AET))-PPTQFC]-OH
A

3BP-4773
N4Ac--PEG6--Nle-[C(3Lut)-PPTEFC]-OH
B

3BP-4775
N4Ac--PPAc--Ttds--Nle--[C(3Lut)-PPTEFC]-OH
B

3BP-4778
nBu--CAyl--[C(tMeBn(NOPO--AET))-PPTEFC]-OH
A

3BP-4784
N4Ac--PPAc--Ttds--Nle--[C(3MeBn)-PPTEFC]-Bal-OH
A

3BP-4844
nBu--CAyl--[C(tMeBn(LuDOTA--PP))-PPTQFC]-NH2
A

3BP-4960
N4Ac--PPAc--Ttds--Nle--[C(3MeBn)-PPTQFC]-Bal-OH
A

3BP-4961
N4Ac--PPAc--Ttds--Nle--[C(3Lut)-PPTQFC]-Bal-OH
A

3BP-5201
NOTA--Ttds--Nle--[C(3MeBn)-PPTQFC]-OH
A

3BP-5210
nBu--CAyl--[C(tMeBn(NOTA--AET))-PPTQFC]-OH
A

Example 45: ¹¹¹In-Labeling of Selected Compounds

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

For the analysis of radiochemical purity by HPLC, 5 μl of diluted labeling solution was analyzed with a Poroshell SB-C18 2.7 μm (Agilent). Eluent A: H₂O, 0.1% TFA, eluent B: MeCN, 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 with the dead volume represents free radionuclide, the peak eluting with the peptide-specific retention time as determined with an unlabeled sample represents radiolabeled compound. Radiochemical purity was ≥85% at end of synthesis. Exemplary radiochemical purities for ¹¹¹In-labeled compounds are shown in Table 29.

TABLE 29

Radiochemical purity by HPLC of ¹¹¹In-labeled compounds.

HPLC
HPLC Area %
HPLC Area %

retention
at end
appr. 4 h post end

time [min]
of synthesis
of synthesis

¹¹¹In-3BP-4560
6.8
88.1
85.8

Example 46: ^99mTc-Labeling of Selected Compounds

100-500 MBq of ^99mTcO₄(in saline) were mixed with 11 μl of 0.5 M phosphate buffer pH 11.5-12.0 and 1.1 μl of 0.1 M trisodium citrate per 100 μl radionuclide solution. 1 nmol of compound (200 μM stock solution in water) per 45 MBq was added, followed by 3.0 μl of 1.1 mg/ml tin chloride dihydrate in nitrogen-purged absolute ethanol per 100 μl radionuclide solution. The mixture was incubated for 30 min at 25-50′° C. At the end of the incubation time, the mixture was neutralized with 3.0 μl 1 M HCl per 10 μl phosphate buffer and TWEEN-20 was added at a final concentration of 0.100.

For the analysis of radiochemical purity by HPLC, 5 μl of diluted labeling solution was analyzed with a Poroshell SB-C18 2.7 μm (Agilent). Eluent A: H₂O, 0.1% TFA, eluent B: MeCN, gradient from 500 B to 70% B within 15 min, flow rate 0.5 ml/min; detector: NaI (Raytest), DAD 230 nm. The peak eluting with the dead volume represents free radionuclide, the peak eluting with the peptide-specific retention time as determined with an unlabeled sample represents radiolabeled compound. Radiochemical purity was ≥85% at end of synthesis. Exemplary radiochemical purities for ^99mTc-labeled compounds are shown in Table 30.

TABLE 30

Radiochemical purity by HPLC of 99Tc-labeled compounds.

HPLC
HPLC Area %
HPLC Area %

retention
at end
appr. 4 h post end

time [min]
of synthesis
of synthesis

3BP-4219
7.8
94.8
93.8
(5.5 h)

3BP-4221
8.2
99.0
97.4
(5.5 h)

3BP-4533
6.7
99.3
98.2
(3 h)

3BP-4534
7.6
98.6
97.5
(3 h)

3BP-4541
7.7
95.8
89.7

3BP-4549
6.5
99.3
97.6
(3 h)

3BP-4550
6.7
100
98.0
(3 h)

3BP-4551
8.2
94.1
90.8

3BP-4552
8.2
88.3
N/A

3BP-4773
6.8
95.8
N/A

3BP-4774
6.3
88.8
86.1

3BP-4775
6.5
86.4
88.1

3BP-4780
8.3
95.0
92.3

3BP-4782
7.8
97.1
95.8

3BP-4784
7.7
96.6
94.9

3BP-4785
7.7
95.3
93.8

3BP-4960
7.6
95.2
92.3

3BP-4961
6.2
98.0
N/A

N/A = not available

Example 47: ⁶⁸Ga-Labeling of Selected Compounds

750 μl Ga-68 eluate (200-500 MBq; 0.1 M HCl) was mixed with 750 μl labeling buffer (1.0 M ammonium acetate buffer pH 4 or 1.0 M ammonium acetate buffer/0.125 M ascorbic acid 4:1 pH 4). 400 μl 50% EtOH was added and the mixture preheated. The activity was measured, and the appropriate amount of peptide (200 μM stock solution in 0.1 M HEPES) was added to reach the desired molar activity (20 MBq/nmol). The mixture was heated for 15 min at 80° C. (3BP-4713, 3BP-4714, 3BP-4724) or 40° C. (3BP-4768 or 3BP-4778, 3BP-5201, 3BP-5210). At the end of the incubation time, the reaction mixture was diluted with 10 ml water before transfer onto a pre-conditioned (5 ml absolute ethanol followed by 10 ml water) Oasis HLB plus light cartridge. The column was washed with 2 ml water and the product eluted in 250 μl absolute ethanol. The final product was formulated in 0.9% sterile NaCl or 0.9% sterile NaCl containing 10 mg/ml ascorbic acid pH 7 to a final concentration of 100 MBq and 10 nmol per ml and a final ethanol concentration of <9%. A sample was withdrawn for determination of radiochemical purity immediately and after final injection.

For the analysis of radiochemical purity by HPLC, 5 μl of diluted labeling solution was analyzed with a XBridge C18 3.5 μM 4.6×50 mm column (Waters). Eluent A: H₂O, 0.1% TFA, eluent B: MeCN, 0.1% TFA; gradient from 5% B to 20% B within 1 min, then 20% B to 50% B within 7 min, flow rate 1.5 ml/min; detector: NaI (Raytest), DAD 220 nm. The peak eluting with the dead volume represents free radionuclide, the peak eluting with the peptide-specific retention time as determined with an unlabeled sample represents radiolabeled compound. Radiochemical purity was ≥95% at end of synthesis. Exemplary radiochemical purities for ⁶⁸Ga-labeled compounds are shown in Table 31.

TABLE 31

Radiochemical purity by HPLC of ⁶⁸Ga-labeled compounds.

HPLC
HPLC
HPLC

retention
Area % after
Area % after

time [min]
formulation
final injection

⁶⁸Ga-3BP-4713
4.6
98.9
98.2

⁶⁸Ga-3BP-4714
4.8
99.2
96.3

⁶⁸Ga-3BP-4724
4.5
98.3
94.0

⁶⁸Ga-3BP-4768
3.8
97.5
96.8

⁶⁸Ga-3BP-4778
4.1
96.2
91.7

⁶⁸Ga-3BP-5201
4.7
99.4
95.9

⁶⁸Ga-3BP-5210
4.2
99.4
91.6

Example 48: In Vivo Imaging Studies

All animal experiments were conducted either in compliance with the German or Denmark animal protection laws. For PET/CT studies, female athymic nude (6- to 8-week-old, Charles River Laboratories, Germany) and for SPECT/CT, swiss nude mice (6- to 8-week-old, Charles River Laboratories, France) were inoculated with 5×10⁶HEK-FAP (embryonic human kidney 293 cells genetically engineered to express high levels of FAP) cells in one shoulder, except for the animals dosed with ¹¹¹In-3BP-4560 which didn't bear any tumor. When tumors reached a size of >150 mm³, mice either received ˜30 MBq of a ^99mTc-labelled, ˜30 MBq ¹¹¹In or ˜10 MBq of a ⁶⁸Ga-labelled compound of the invention (diluted to 100 μL with PBS or saline) administered intravenously via the tail vein. Images were obtained on a nanoScan© SPECT/CT system (Mediso Medical Imaging Systems, Budapest, Hungary) or a nanoScan© PET/CT (Mediso Medical Imaging Systems, Budapest, Hungary) using exemplarily the acquisition and reconstruction parameters listed in Table 32 and 33.

Imaging data were saved as DICOM files and analyzed using either VivoQuant™ (Invicro, Boston, USA) for SPECT/CT or InterView™ FUSION (Mediso, Budapest, Hungary) software for PET/CT. Results are expressed as a percentage of injected dose per gram of tissue (% ID/g).

The results of the imaging studies are presented in tables 34 for SPECT/CT and 35 for PET/CT; acquired scans of selected compounds are shown in FIGS. 22-28

Surprisingly, the modification of the N-terminal linker, to which the N4Ac-chelator was attached, drastically improved the biodistribution of the tracers. In FIGS. 22-25 the representative biodistributions of four ^99mTc-labelled compounds are shown over time (1-6 h post injection). Compounds with a PPAc-Ttds linker between N4Ac and Norleucine (3BP-4541 and -4961) presented decreased uptake in healthy tissues over time, especially in the gastrointestinal tract and the kidneys when compared to compounds with a different linker such as Ttds or PEG6 (3BP-4219 and 3BP-4221).

Additionally, a drastically improved biodistribution of tracers comprising an N-terminal urea motif was found. In FIGS. 21 and 28 the representative biodistributions of the ¹¹¹In-labelled 3BP-3940 and 3BP-4560 are shown over time (0.25-3 h post injection). The tracers presented a very low uptake in healthy tissues, especially in the kidneys.

TABLE 32

Acquisition and reconstruction parameters of SPECT/CT imaging

Acquisition parameters SPECT

System
nanoScan ™ SPECT/CT

Scan range
whole body, 3-bed holder (mouse

hotel)

Time per projection
60 s

Aperture model, pinhole
Aperture #2, 1.5 mm

diameter

Reconstruction parameters

Method
HiSPECT (Scivis), iterative

reconstruction

Smoothing
35%

Iterations
9

Voxel size
0.15 mm × 0.15 mm × 0.15 mm

Acquisition parameters CT

System
NanoSPECT/CT ™

Scan range
whole body, 3-bed holder (mouse

hotel)

Scan duration
7 minutes

Tube voltage
45 kVp

Exposure time
500 ms

Number of projections
240

TABLE 33

Acquisition and reconstruction parameters of PET/CT imaging

Acquisition parameters PET

System
nanoScan ™ PET/CT

Type of scan
static

Scan range
whole body,

3-bed holder (mouse hotel)

Energy window
400-600
keV

List mode
300 s (15 min scan);

600 s (60 min),

900 s (180 min)

Reconstruction
3D maximum a posteriori algorithm

with scatter and attenuation

correction

Acquisition parameters CT

Scan range
whole body,

3-bed holder (mouse hotel)

Type of scan
Helical

Tube voltage
50
kVp

Exposure time
300
ms

Number of projections
480

Isotropic voxel size
250
μm

Binning
1:4

TABLE 34

Shows the percentage of injected dose per gram of tissue (% ID/g)

uptake in the HEK-FAP tumor, kidneys, and BPS (blood pool surrogate)

as determined by SPECT/CT-imaging of ^99mTc-labeled

compounds at 1 h, 3 h, and 6 h post tracer injection.

Tumor
Tumor
Tumor
Kidneys
Kidneys
Kidneys
BPS
BPS
BPS

1 h
3 h
6 h
1 h
3 h
6 h
1 h
3 h
6 h

3BP-4221
8.51
8.32
7.97
4.74
3.73
2.56
0.54
0.27
0.16

3BP-4219
6.80
7.62
4.52
1.85
1.84
0.55
0.21
0.09
0.07

3BP-4533
5.45
6.27
6.75
6.04
4.43
3.04
1.72
1.46
0.86

3BP-4534
5.67
6.85
7.68
2.06
1.40
1.09
1.70
1.19
0.78

3BP-4541
8.81
9.76
7.48
4.02
2.41
1.01
0.48
0.20
0.06

3BP-4549
6.04
4.25
1.68
4.22
1.82
0.58
0.18
0.03
0.01

3BP-4550
6.14
4.91
2.91
0.92
0.62
0.38
0.06
0.03
0.04

3BP-4551
10.56
7.31
4.59
4.34
1.18
0.50
0.18
0.07
0.04

3BP-4552
7.92
6.86
5.21
3.17
3.40
1.34
0.52
0.14
0.07

3BP-4773
6.19
6.19
5.42
6.64
4.71
3.12
0.25
0.09
0.06

3BP-4774
6.14
3.70
1.74
1.87
0.85
0.48
0.08
0.06
0.03

3BP-4775
7.88
6.28
4.55
8.98
6.31
4.28
0.21
0.18
0.04

3BP-4780
6.71
7.01
7.92
8.11
6.14
4.36
0.81
0.61
0.43

3BP-4782
6.20
6.77
7.17
7.28
5.78
4.24
0.69
0.47
0.36

3BP-4784
7.65
7.43
7.44
1.61
1.23
1.01
0.90
0.63
0.38

3BP-4785
4.20
4.14
4.32
3.47
2.14
1.63
0.92
0.64
0.48

3BP-4960
10.54
12.82
12.06
1.96
1.21
1.21
0.74
0.47
0.17

3BP-4961
10.46
11.36
12.30
2.36
1.12
0.77
0.47
0.31
0.11

TABLE 35

Shows the percentage of injected dose per gram of tissue (% ID/g)

uptake in the HEK-FAP tumor, kidneys, and BPS (blood pool

surrogate) as determined by PET/CT-imaging of ⁶⁸Ga-labeled

compounds at 0.25 h, 1 h, and 3 h post tracer injection;

Tumor
Tumor
Tumor
Kidneys
Kidneys
Kidneys
BPS
BPS
BPS

0.25 h
1 h
3 h
0.25 h
1 h
3 h
0.25 h
1 h
3 h

3BP-4713
3.46
3.90
3.21
9.05
2.50
0.82
2.27
0.77
0.19

3BP-4714
5.15
5.44
5.49
5.00
3.82
3.83
1.52
0.65
0.32

3BP-4724
3.87
4.37
4.46
4.28
1.87
0.86
2.17
1.04
0.49

3BP-4768
7.82
9.42
9.20
18.43
2.45
0.67
2.54
0.78
0.27

3BP-4778
3.66
4.06
4.45
3.38
1.81
1.00
1.61
0.77
0.33

3BP-5201
5.56
6.30
6.25
9.27
1.97
1.00
2.31
0.85
0.38

3BP-5210
7.48
8.90
9.53
37.28
5.61
0.42
2.72
0.68
0.14

REFERENCES

The disclosure of each and any document recited herein is incorporated by reference.

COMPOUNDS COMPRISING A FIBROBLAST ACTIVATION PROTEIN LIGAND AND USE THEREOF

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