Radionuclide-labeled peptide conjugates for site-specific uPAR targeting

Abstract

The present invention provides radionuclide-labeled peptides for site-specific targeting of urokinase plasminogen activator receptor (uPAR) and treatment of cancer diseases associated with high uPAR expression.

Description

[Technical Field]

[0001] The present invention relates to radionuclide-labeled peptides for site-specific targeting of urokinase plasminogen activator receptor (uPAR) and the treatment of cancer diseases associated with high uPAR expression. [Background technology]

[0002] For the site-specific targeting of therapeutic radionuclides, various radionuclide-labeled peptide compositions have been developed or are under development. The general principle involves binding a selected radionuclide to a peptide with high specificity for a particular organ or tissue, so that the organ or tissue can be treated with the therapeutic radioisotope. This research area has shown promising applicability in tumor treatment.

[0003] Malignant tumors have the ability to degrade the surrounding extracellular matrix, leading to local invasion or metastasis. Urokinase-type plasminogen activator (uPA) and its cell surface receptor (uPAR) are central molecules in cell surface-associated plasminogen activation both in vitro and in vivo. While uPAR is overexpressed in various human cancers, its expression in non-cancerous tissues is low. It correlates with malignant tumor growth and is associated with poor prognosis in many types of human cancer, possibly suggesting a causal role for the uPA / uPAR system in cancer progression and metastasis. Immunohistochemistry and in situ hybridization studies have shown that expression levels of components derived from the uPA system are generally very low in normal tissues and benign lesions. The uPA / uPAR system has also been reported to be involved in regulating cell-extracellular matrix interactions by acting as an adhesion receptor for vitronectin and regulating integrin function. Based on these properties, the uPA / uPAR system has consequently been considered an attractive target for cancer therapy, and uPAR-binding peptides, including the peptide AE105, have been developed to target the uPA / uPAR system, as described in U.S. Patent No. 7,026,282. Furthermore, as described in International Publication No. WO 2013 / 167130, a radionuclide-labeled uPAR-targeting peptide conjugate, 177Lu-DOTA-AE105, was developed in pursuit of providing targeted radiotherapy for uPAR-expressing cancers. However, tumor binding and tumor retention of this peptide conjugate were significantly reduced compared to that of the peptide alone, indicating that the AE105 peptide is not well suited for labeling with therapeutic radionuclides and has limited potential for providing targeted radiotherapy.

[0004] The development of therapeutic radionuclide-labeled peptide conjugates is a complex process that poses various challenges, including conjugate stability, which can be compromised by, for example, radiolysis by the therapeutic radionuclide or insufficient binding of the radionuclide within the chelator in a given conjugate structure. Furthermore, particularly for therapeutic applications, target specificity is crucial for achieving low toxicity and therapeutic efficacy. Conjugate retention at the target is also crucial for achieving optimal therapeutic efficacy. Despite extensive research into peptide conjugates targeting various molecules, only two such conjugates have been approved for therapeutic use: 177Lu-DOTA-TATE (Lutathera) and 177Lu-DOTA-PSMA (Pluvicto) for the treatment of hormone receptor somatostatin-positive gastrointestinal pancreatic neuroendocrine tumors (GEP-NETs) and metastatic castration-resistant prostate cancer (PSMA-positive mCRPC). Therefore, there is a need for stable radionuclide-labeled uPAR-targeting conjugates that provide sufficient tumor binding and good tumor retention so that cancer diseases associated with high uPAR expression can be treated with targeted radiotherapy. Summary of the Invention

[0005] Surprisingly, the inventors found that the Cu-DOTA-AE105 conjugate exhibited approximately 50-fold higher tumor binding and much longer tumor retention than Lu-DOTA-AE105 (see Example 5). Furthermore, Cu-DOTA-AE105 was found to be therapeutically effective in preclinical studies in tumor mouse models, whereas Lu-DOTA-AE105 did not exhibit therapeutic efficacy in the same tumor mouse models (see Example 4). Cu-DOTA-AE105 also demonstrated therapeutic efficacy in two additional preclinical studies in tumor mouse models (see Examples 6 and 7), confirming that uPAR-targeting peptide-based radionuclide conjugates, and in particular Cu-DOTA-AE105, are generally therapeutically effective in various types of cancers that express uPAR. Furthermore, the inventors found that the 225Ac-DOTA-AE105 conjugate exhibited approximately five-fold higher tumor binding compared to 177Lu-DOTA-AE105, demonstrated therapeutic efficacy in a glioblastoma xenograft tumor mouse model, and showed promising results in a tolerability study. Thus, although previous studies had dismissed the AE105 peptide as a suitable platform for future development of targeted radiotherapy for uPAR-expressing cancers, the inventors have successfully developed therapeutically effective radionuclide-labeled uPAR-binding conjugates based on the AE105 peptide, in conjunction with both alpha- and beta-emitting radionuclides.

[0006] The radionuclide-labeled peptide conjugates of the present invention have high tumor binding and good tumor retention, and offer a much broader therapeutic window than inferior counterparts, allowing for achieving doses that provide an optimal balance of therapeutic efficacy and side effects.

[0007] In a first aspect, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis, comprising: a) uPAR-binding peptide b) Chelating agents suitable for binding radioactive metals c) comprising a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb, and Tb; The peptide is conjugated to a radionuclide by a chelating agent. Radionuclide-labeled uPAR-binding peptide conjugates.

[0008] The present invention provides an excellent radionuclide-labeled uPAR-binding peptide conjugate suitable for the treatment of uPAR-expressing cancer diseases.

[0009] For the sake of completeness, it is hereby made clear that the radionuclide-labeled uPAR-binding peptide conjugate according to the first aspect and all its embodiments preferably consists of the specified components. That is, in a preferred form of the first aspect, the present invention provides a) uPAR-binding peptide b) Chelating agents suitable for binding radioactive metals c) consisting of a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb, and Tb; the peptide is conjugated to said radionuclide by a chelating agent; Radionuclide-labeled uPAR-binding peptide conjugates.

[0010] In a second aspect, the present invention relates to a radionuclide-labelled uPAR-binding peptide conjugate according to the first aspect for use as a medicament.

[0011] In a third aspect, the present invention relates to a radionuclide-labelled uPAR-binding peptide conjugate according to the first aspect for use in the treatment of a uPAR-expressing cancer.

[0012] In a fourth aspect, the present invention relates to a method for treating a uPAR-expressing cancer disease by administering to a patient a radionuclide-labeled uPAR-binding peptide conjugate according to the first aspect. [Brief explanation of the drawings]

[0013] [Figure 1] FIG. 1 shows the biodistribution of 67Cu-DOTA-AE105 in xenograft tumor (U87MG)-bearing mice. [Figure 2]SPECT / CT images of the biodistribution of Cu-DOTA-AE105 in mice bearing human xenograft tumors (U87MG). More specifically, Figure 2a shows axial, coronal, and maximum intensity projection (MIP) images of a representative animal from Group A (30 MBq), and Figure 2b shows axial, coronal, and maximum intensity projection (MIP) images of a representative animal from Group B (60 MBq). [Figure 3] FIG. 1 shows the binding specificity of 67Cu-DOTA-AE105 in uPAR-expressing tumors. [Figure 4] FIG. 1 shows the treatment efficacy of 67Cu-DOTA-AE105 in a glioblastoma mouse model. [Figure 5] Figure 1 shows Kaplan-Meier curves of survival data from a treatment efficacy study of 67Cu-DOTA-AE105 in a glioblastoma mouse model. [Figure 6] FIG. 1 shows the treatment efficacy of 177Lu-DOTA-AE105 in a human xenograft glioblastoma (U87MG) mouse model. [Figure 7] FIG. 1 shows the treatment efficacy of 67Cu-DOTA-AE105 in a human xenograft glioblastoma mouse model (U87.MG) and a human xenograft non-small cell lung cancer mouse model (NCI-1993). [Figure 8] FIG. 1 shows the therapeutic effect of 67Cu-DOTA-AE105 in a human xenograft colorectal cancer mouse model. [Figure 9] FIG. 1 shows 67Cu-DOTA-AE105-induced apoptosis and DNA double-strand breaks in the treatment of a human xenograft colorectal cancer mouse model. [Figure 10] FIG. 1 shows the biodistribution and binding specificity of 225Ac-DOTA-AE105 in mice bearing human xenograft tumors (U87MG). [Figure 11] FIG. 1 shows the tolerability of 225Ac-DOTA-AE105. [Figure 12a]FIG. 1 shows a representation of the HPLC chromatogram of 30 kBq / nmol 225Ac-DOTA-AE105 based on gamma-counted HPLC fractions. [Figure 12b] FIG. 1 shows a representation of the HPLC chromatogram of 40 kBq / nmol Ac-225-DOTA-AE105 based on gamma-counted HPLC fractions. [Figure 13] FIG. 2 shows 225Ac-DOTA-AE105-induced apoptosis in glioblastoma xenograft tumors. DETAILED DESCRIPTION OF THE INVENTION

[0014] The radionuclide-labeled uPAR-binding peptide conjugate according to the first aspect of the present invention, which comprises a uPAR-binding peptide, a chelating agent suitable for binding a radioactive metal, and a radionuclide selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, wherein the peptide is bound to the radionuclide via the chelating agent, is a stable conjugate structure with high specificity for uPAR and good retention capacity, thereby enabling the treatment of uPAR-expressing cancers, particularly cancer types in which uPAR is highly expressed.

[0015] uPAR-binding peptide The uPAR-binding peptide of the present invention can be a monomeric peptide, i.e., a single peptide, or a multimeric peptide construct, such as a dimer, trimer, or tetramer.The multimeric peptide construct can be a homomer (i.e., consisting of identical peptides) or a heteromer (i.e., consisting of non-identical uPAR-binding peptides), and the multimeric peptide construct can have a higher affinity for target receptors than its monomeric counterpart.

[0016] The uPAR-binding peptides of the present invention can be synthesized using standard methods. An example of a standard method is described in U.S. Patent No. 7,026,282 as follows: The chain elongation step of solid-phase peptide synthesis was performed manually using a polyethylene syringe as a reaction vessel. Synthesis was performed on KA resin with Fmoc-amino acids preloaded onto an acid-labile linker. Five equivalents of Fmoc-amino acids activated with 1-hydroxy-7-benzotriazole (HOBt) and N,N'-diisopropylcarbodiimide were used in the coupling step, and the reaction was allowed to proceed for over two hours. The Fmoc protecting group was removed with 20% piperidine in dimethylformamide for 15-20 minutes. For cleavage, the peptide resin was treated with 85% TFA containing 5% each of phenol, mercaptoethanol, and thioanisole for 1.5 hours. The filtrate was concentrated by nitrogen flushing, and then the peptide was precipitated from diethyl ether and washed four times with diethyl ether. Finally, the peptides were dissolved / suspended in glacial acetic acid, lyophilized, redissolved in 10% acetic acid, and lyophilized again. Analytical HPLC analysis was performed on a Cs column using a Waters 600E equipped with a Waters photodiode array detector. A 25-minute linear gradient from Buffer A (0.1% TFA, 9.9% HO, 90% CHCN) was used. If necessary, peptides were purified on a preparative scale. The exact identity of the peptides was confirmed by matrix-assisted laser desorption / ionization mass spectrometry or electrospray ionization mass spectrometry. Purity was confirmed by reverse-phase HPLC.

[0017] "Multimeric" peptides can be synthesized by standardized peptide synthesis methodologies using orthogonal protection strategies (e.g., those described in Cwirla et al., Science, Vol. 276, 1997, pp. 1696-1699). Alternatively, heterogeneous methods well known to peptide chemists for the construction of multimeric antigens, e.g., for immunization (DN Posnett et al., (1988) J. Biol. Chem. 263:1719-1725), are also applicable within the present invention.

[0018] In one embodiment of the first aspect of the invention, the uPAR-binding peptide is a monomeric or multimeric peptide, wherein one or more of the peptide components is selected from the group consisting of: (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile).

[0019] In a preferred embodiment of the first aspect of the invention, the uPAR-binding peptide is a monomeric peptide selected from the group consisting of: (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile).

[0020] All of these peptides share a minimal structure by possessing at least a portion of the binding hotspot of x-Cha-Phe-xxx-Leu-Trp-x, where the bolded residues are the hotspot for interaction with uPAR, Cha is cyclohexyl-(L)alanine, and x represents any amino acid. These peptides themselves possess high specificity and good affinity for the target uPAR, allowing the resulting radionuclide-labeled uPAR-binding peptide conjugates to be sufficiently retained at the target for sufficient time for the radionuclide to provide therapeutically relevant radiation exposure of the target area. Furthermore, the peptides offer good compatibility with appropriate chelators, including DOTA, DOTAM, NOTA, NODAGA, sarcofadin, CB-TE2A, and their derivatives, especially DOTA.

[0021] In a most preferred embodiment of the first aspect of the present invention, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), also referred to in the context of the present invention as AE105. This provides a uPAR-binding peptide that itself has excellent specificity as well as high affinity for the target uPAR, thereby promoting excellent retention of the radionuclide-labeled uPAR-binding peptide conjugate at the target, allowing sufficient time for the radionuclide to provide therapeutically relevant radiation exposure of the target area. Furthermore, the uPAR-binding peptides offer good compatibility with appropriate chelators, including DOTA, DOTAM, NOTA, NODAGA, sarcofadin, CB-TE2A and their derivatives, especially DOTA, and provide excellent target binding ability when labeled with selected radionuclides 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, especially 67Cu, 212Pb and 225Ac.

[0022] Radiolabeling of one or more peptides selected for the radionuclide-labeled uPAR-binding peptide conjugate of the present invention is made possible by coupling a suitable chelator capable of binding a radiometal to the peptide, the chelator being capable of binding the selected radionuclide.

[0023] chelating agents The use of various chelating agents suitable for binding radiometals for radiolabeling peptides is well known in the art.Suitable chelating agents generally include those containing tetradentate ligands with at least one sulfur group available for binding to metal radionuclides, such as known N3S and N2S2 ligands.The chelating agent is attached to the uPAR-binding peptide by standard methodologies well known in the field of the present invention, and can be added to any position on the uPAR-binding peptide, as long as the biological activity, such as the binding properties and specificity of the peptide, is not adversely affected.Preferably, the chelating group is covalently attached to the amino-terminal amino acid of the peptide.The chelating group can be advantageously attached to the peptide during solid-phase peptide synthesis, or added by solution-phase chemistry after the peptide is obtained. Preferred chelating agents include DOTA (1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecane), DOTAM (2-[4,7,10-tris(2-amino-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl]acetamide), NOTA (2,2',2''-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid), ), NODAGA, sarcofagin (3,6,10,13,16,19-hexazabicyclo(6,6,6)icosane), CB-TE2A (1,4,8,11-tetraazabicyclo[6.6.2]hexadecane-4,11-diacetic acid) and their derivatives, which constitute an important class of chelating agents for biomedical applications because they accommodate a variety of divalent and trivalent metal ions with great stability.

[0024] DOTA (dodecanetetraacetic acid) and its derivatives, which have the chemical formula (1,4,7,10-tetrakis(carboxymethyl)-1,4,7,10-tetraazacyclododecane), are particularly recognized to constitute an important class of chelating agents for biomedical applications, as they accommodate a variety of divalent and trivalent metal ions, such as 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, with great stability.

[0025] DOTAM, also known as TCMC, is a DOTA analogue.

[0026] Sarcofazine (Sar) is a bicyclic, cage-shaped chelating molecule derived from cyclam. Its chemical formula is 3,6,10,13,16,19-hexazabicyclo(6,6,6)icosane; additional functional or non-functional groups are often attached to this structure, resulting in derivatives such as DiAmSar (1,8-diamino-Sar), AmBaSar (4-((8-amino-3,6,10,13,16,19-hexazabicyclo[6.6.6]icosan-1-ylamino)methyl)benzoic acid), and MeCOSar (5-(8-methyl-3,6,10,13,16,19-hexaaza-bicyclo[6.6.6]icosan-1-ylamino)-5-oxopentanoic acid). Sarcofazine and its derivatives are suitable chelating agents for use in the radionuclide-labeled uPAR-binding peptide conjugates of the present invention, and are preferred for the 67Cu-labeled uPAR-binding peptide conjugates of the present invention.

[0027] In one embodiment of the first aspect of the present invention, the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, sarcofaddin, CB-TE2A and derivatives thereof, preferably DOTA, DOTAM, NOTA, NODAGA, sarcofaddin, DiAmSar, AmBaSar, MeCOSar or CB-TE2A.

[0028] In a preferred embodiment of the first aspect of the present invention, the chelating agent is selected from the group consisting of DOTA, DOTAM, sarcofazine, and derivatives thereof, preferably DOTA, DOTAM, sarcofazine, DiAmSar, AmBaSar, or MeCOSar.

[0029] In a more preferred embodiment of the first aspect of the invention, the chelating agent is DOTA.

[0030] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is DOTA and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac, or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0031] In one embodiment of the first aspect of the invention, the chelator is DOTA and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac, or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0032] In a preferred embodiment of the first aspect of the invention, the chelator is DOTA, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0033] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is DOTAM and the radionuclide is selected from the group consisting of Cu, Ac, Pb, Tb, and Tb, preferably Cu, Ac, or Pb, more preferably Cu or Ac, and most preferably Cu.

[0034] In one embodiment of the first aspect of the invention, the chelating agent is DOTAM and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0035] In a preferred embodiment of the first aspect of the invention, the chelating agent is DOTAM, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0036] In another preferred embodiment of the first aspect of the invention, the chelating agent is DOTAM, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is Pb.

[0037] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is NOTA and the radionuclide is selected from the group consisting of Cu, Ac, Pb, Tb and Tb, preferably Cu, Ac or Pb, more preferably Cu or Ac, and most preferably Cu.

[0038] In one embodiment of the first aspect of the invention, the chelator is NOTA and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0039] In a preferred embodiment of the first aspect of the invention, the chelator is NOTA, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0040] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is NODAGA and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0041] In one embodiment of the first aspect of the invention, the chelating agent is NODAGA and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac, or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0042] In a preferred embodiment of the first aspect of the invention, the chelating agent is NODAGA, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0043] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is MeCOSar and the radionuclide is selected from the group consisting of Cu, Ac, Pb, Tb and Tb, preferably Cu, Ac or Pb, more preferably Cu or Ac, and most preferably Cu.

[0044] In one embodiment of the first aspect of the invention, the chelator is MeCOSar and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac, or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0045] In a preferred embodiment of the first aspect of the invention, the chelating agent is MeCOSar, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0046] In one embodiment of the first aspect of the invention, the chelator of the uPAR-binding peptide conjugate is CB-TE2A and the radionuclide is selected from the group consisting of Cu, Ac, Pb, Tb and Tb, preferably Cu, Ac or Pb, more preferably Cu or Ac, and most preferably Cu.

[0047] In one embodiment of the first aspect of the invention, the chelator is CB-TE2A and the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile); The radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac, or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0048] In a preferred embodiment of the first aspect of the invention, the chelator is CB-TE2A, the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), and the radionuclide is selected from the group consisting of 67Cu, 225Ac, 212Pb, 161Tb, and 149Tb, preferably 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, and most preferably 67Cu.

[0049] radionuclides The group of radionuclides according to the first aspect of the invention are characterized by undergoing either beta-minus or alpha decay, exposure to which is damaging to cells, for example, in the human body. Therefore, these radionuclides (Cu, Ac, Pb, Tb, and Tb) are generally considered to be therapeutic radionuclides particularly suitable for the treatment of cancer.

[0050] The peptide / chelator conjugate of the first aspect of the invention is labeled by reacting the conjugate with a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb, and Tb, e.g., as a metal salt, preferably water-soluble. The reaction is carried out by methods well known in the art.

[0051] Preferred radionuclides for labelling the uPAR-binding peptide conjugate of the first aspect of the invention are 67Cu, 225Ac or 212Pb, more preferably 67Cu or 225Ac, most preferably 67Cu.

[0052] 67Cu is most often found in its divalent state. 67Cu is β -It decays by (beta minus) decay. The half-life of 67Cu is 2.6 days, making it particularly suitable for therapeutic use in targeted cancer therapy. In connection with the present invention, the inventors have found that 67Cu exhibits surprisingly good compatibility with the uPAR-binding peptide conjugates of the present invention, providing stably labeled conjugates with high specificity for their targets, thereby demonstrating that beta-emitting radionuclides, particularly 67Cu, are suitable labeling agents for the conjugates of the present invention.

[0053] In one embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is DOTAM.

[0054] In one embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is NOTA.

[0055] In one embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is NODAGA.

[0056] In one embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is sarcofazine.

[0057] In one embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is CB-TE2A.

[0058] In a preferred embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is DOTA or MeCOSar.

[0059] In a more preferred embodiment of the first aspect of the present invention, the chelator in the 67Cu-labeled uPAR-binding peptide conjugate of the present invention is DOTA.

[0060] In a further most preferred embodiment of the first aspect of the present invention, the radionuclide-labeled uPAR-binding peptide conjugate has the formula: [ka] It has.

[0061] 225Ac is an actinide, most often in a trivalent state. 225Ac decays by alpha decay with a half-life of 9.9 days. This makes it particularly suitable for therapeutic use in targeted cancer therapy. In connection with the present invention, the inventors have found that 225Ac exhibits good compatibility with the uPAR-binding peptide conjugate of the present invention, providing a stably labeled conjugate with high specificity for its target, thereby demonstrating that alpha-emitting radionuclides, particularly 225Ac, are suitable labeling agents for the conjugates of the present invention.

[0062] In one embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is DOTAM.

[0063] In one embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is NOTA.

[0064] In one embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is NODAGA.

[0065] In one embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is sarcofazine.

[0066] In one embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is CB-TE2A.

[0067] In a preferred embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is DOTA or MeCOSar.

[0068] In a more preferred embodiment of the first aspect of the present invention, the chelator in the 225Ac-labeled uPAR-binding peptide conjugate of the present invention is DOTA.

[0069] In an even more preferred embodiment of the first aspect of the present invention, the radionuclide-labeled uPAR-binding peptide conjugate has the formula: [ka] It has.

[0070] 212Pb (212 lead) decays by beta-minus decay with a half-life of 11 hours to produce 212Bi (212 bismuth), which emits alpha rays. This provides a dual therapeutic effect (i.e., beta-minus and alpha emissions are induced), making it particularly suitable for therapeutic applications in targeted cancer treatment. In the context of the present invention, the inventors have found that Pb exhibits good compatibility with the uPAR-binding peptide conjugates of the present invention, providing conjugates with high affinity for their targets, which is an important prerequisite for the therapeutic use of 212Pb-labeled conjugates.

[0071] In one embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is DOTAM.

[0072] In one embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is NOTA.

[0073] In one embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is NODAGA.

[0074] In one embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is sarcofazine.

[0075] In one embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is CB-TE2A.

[0076] In a preferred embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is DOTA or MeCOSar.

[0077] In a more preferred embodiment of the first aspect of the present invention, the chelator in the 212Pb-labeled uPAR-binding peptide conjugate of the present invention is DOTA.

[0078] In an even more preferred embodiment of the first aspect of the present invention, the radionuclide-labeled uPAR-binding peptide conjugate has the formula: [ka] It has.

[0079] 161Tb is a radioactive lanthanide, most often in the trivalent state. 161Tb decays by beta-minus decay with a half-life of 7 days. This makes it suitable for therapeutic use, particularly in targeted cancer therapy.

[0080] In one embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTAM.

[0081] In one embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is NOTA.

[0082] In one embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is NODAGA.

[0083] In one embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is sarcofazine.

[0084] In one embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is CB-TE2A.

[0085] In a preferred embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTA or MeCOSar.

[0086] In a most preferred embodiment of the first aspect of the present invention, the chelator in the 161Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTA.

[0087] 149Tb is a radioactive lanthanide, most often in the trivalent state. 149Tb decays by alpha decay with a half-life of 5.9 days. This makes it suitable for therapeutic use, particularly in targeted cancer therapy.

[0088] In one embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTAM.

[0089] In one embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is NOTA.

[0090] In one embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is NODAGA.

[0091] In one embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is sarcofazine.

[0092] In one embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is CB-TE2A.

[0093] In a preferred embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTA or MeCOSar.

[0094] In a most preferred embodiment of the first aspect of the present invention, the chelator in the 149Tb-labeled uPAR-binding peptide conjugate of the present invention is DOTA.

[0095] disease The present inventor has demonstrated therapeutic effect in three different uPAR-expressing cancer diseases.It is generally recognized that therapeutic effect across two or three different cancer diseases is very strong support for the therapeutic effect of cancers that generally express related targets.Therefore, the radionuclide-labeled uPAR-binding peptide conjugate of the present invention can treat uPAR-expressing cancer, particularly the cancer types with high uPAR expression, such as breast cancer, brain cancer, particularly high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), SCLC (small cell lung cancer), neuroendocrine tumor, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer) and gastric cancer.

[0096] One embodiment of the third aspect of the present invention relates to a radionuclide-labelled uPAR-binding peptide conjugate of the first aspect of the present invention for use in the treatment of breast cancer, brain cancer, in particular high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), SCLC (small cell lung cancer), neuroendocrine tumours, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer) and gastric cancer, preferably breast cancer, NSCLC, neuroendocrine tumours, HNSCC, pancreatic cancer, prostate cancer, CRC, glioblastoma and gastric cancer, more preferably NSCLC, CRC and glioblastoma, most preferably glioblastoma.

[0097] One embodiment of the fourth aspect of the present invention relates to a method for treating a uPAR-expressing cancer disease by administering to a patient a radionuclide-labeled uPAR-binding peptide conjugate according to the first aspect, wherein the cancer disease is selected from breast cancer, brain cancer, in particular high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), SCLC (small cell lung cancer), neuroendocrine tumors, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer) and gastric cancer, preferably NSCLC, neuroendocrine tumors, HNSCC, pancreatic cancer, prostate cancer, CRC, glioblastoma and gastric cancer, more preferably NSCLC, CRC and glioblastoma, most preferably glioblastoma.

[0098] Radionuclide-labeled uPAR-binding peptide conjugates of the present invention, in which the radionuclide is a beta-emitter, e.g., Cu and Tb, are prepared to administer to an individual a radiation dose of about 1-100 MBq, preferably about 30-60 MBq, in animals, and 1-20 GBq, preferably about 2-10 GBq, and most preferably about 7.4 GBq, in humans, according to standard radiopharmaceutical dosage determinations. This dose may be administered in one to four increments. Radionuclide-labeled uPAR-binding peptide conjugates of the present invention, in which the radionuclide is an alpha-emitter, e.g., Ac, Pb, or Tb, are prepared to administer to an individual a radiation dose of about 1-100 KBq, preferably about 5-60 KBq, in animals, 1-300 MBq, preferably about 2-10 MBq, in humans, and 50-200 MBq in Pb, according to standard radiopharmaceutical dosage determinations. This dose may be administered in 1 to 4 increments. The radiolabeled peptide may be administered intravenously in a conventional vehicle for intravenous injection.

[0099] List of embodiments Embodiment 1, a radionuclide-labeled uPAR-binding peptide conjugate, comprising: a) uPAR-binding peptide b) a chelating agent suitable for binding radiometals c) a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb and Tb; The peptide is conjugated to a radionuclide by a chelating agent. Radionuclide-labeled uPAR-binding peptide conjugates.

[0100] Preferably, the radionuclide-labeled uPAR-binding peptide conjugate comprises: a) uPAR-binding peptide b) a chelating agent suitable for binding radiometals c) a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb and Tb; The peptide is conjugated to a radionuclide by a chelating agent.

[0101] Embodiment 2. The radionuclide-labeled peptide conjugate of embodiment 1, wherein the uPAR-binding peptide is a monomeric or multimeric peptide, and one or more peptide components are selected from the group consisting of: (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile).

[0102] Embodiment 3. The radionuclide-labeled peptide conjugate of any one of embodiments 1 or 2, wherein the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser).

[0103] Embodiment 4: The radionuclide-labeled uPAR-binding peptide conjugate of any one of embodiments 1 to 3, wherein the chelator is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, sarcofadin, CB-TE2A, or derivatives thereof.

[0104] Embodiment 5. The radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 4, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, and sarcofazine.

[0105] Embodiment 6. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 5, wherein the chelating agent is DOTA.

[0106] Embodiment 7. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 6, wherein the radionuclide is 67Cu, 225Ac or 212Pb, preferably 67Cu or 225Ac.

[0107] Embodiment 8. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 67Cu.

[0108] Embodiment 9, the formula: [ka] 9. The radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 8, comprising:

[0109] Embodiment 10: A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 225Ac.

[0110] Embodiment 11, the formula: [ka] 11. The radionuclide-labeled uPAR-binding peptide conjugate of any one of embodiments 1 to 7 or 10, having the formula:

[0111] Embodiment 12. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 212Pb.

[0112] Embodiment 13. The formula: [ka] 13. The radionuclide-labeled uPAR-binding peptide conjugate of any one of embodiments 1 to 7 or 12, having the formula:

[0113] Embodiment 14. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 67Cu and the chelator is DOTA.

[0114] Embodiment 15. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 67Cu and the chelating agent is sarcophagin.

[0115] Embodiment 16. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 212Pb and the chelator is DOTAM.

[0116] Embodiment 17. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 7, wherein the radionuclide is 67Cu and said chelator is CB-TE2A.

[0117] Embodiment 18: A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 17 for use as a medicament.

[0118] Embodiment 19. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 17 for use in treating a uPAR-expressing cancer.

[0119] Embodiment 20: The radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 17, for use according to claim 19, wherein the cancer is selected from the group consisting of breast cancer, brain cancer, in particular high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), SCLC (small cell lung cancer), neuroendocrine tumors, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer), and gastric cancer.

[0120] Embodiment 21: The radionuclide-labeled uPAR-binding peptide conjugate of any one of embodiments 1 to 17, for use according to embodiment 19 or embodiment 20, wherein the cancer is selected from the group consisting of breast cancer, NSCLC, neuroendocrine tumors, HNSCC, pancreatic cancer, prostate cancer, CRC, glioblastoma and gastric cancer, preferably NSCLC, CRC and glioblastoma.

[0121] Embodiment 22: The radionuclide-labeled uPAR-binding peptide conjugate of any one of embodiments 1 to 17, for use according to any one of embodiments 19 to 21, wherein the cancer is selected from the group consisting of glioblastoma.

[0122] Embodiment 23. A method for treating uPAR-expressing cancer diseases by administering to a patient a radionuclide-labeled uPAR-binding peptide conjugate according to any one of embodiments 1 to 17.

[0123] Embodiment 24: The method according to embodiment 23, wherein the cancer disease is selected from breast cancer, brain cancer, particularly high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), sclc (small cell lung cancer), neuroendocrine tumor, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer), and gastric cancer.

[0124] Embodiment 25: The method according to embodiment 23 or 24, wherein the cancer disease is selected from NSCLC, neuroendocrine tumor, HNSCC, pancreatic cancer, prostate cancer, CRC, glioblastoma and gastric cancer, preferably NSCLC, CRC and glioblastoma.

[0125] Embodiment 26: The method of any one of embodiments 23 to 25, wherein the cancer disease is glioblastoma. [Example]

[0126] Example 1: Radiolabel 67Cu labeling of DOTA-AE105 conjugate A radionuclide-labeled uPAR-binding peptide conjugate according to the present invention was prepared by 67Cu-labeling of a DOTA-AE105 conjugate.

[0127] The compounds were labeled with molar radioactivities of 30 and 60 MBq / nmol.

[0128] The desired amount of radioactivity from the 67Cu stock solution (in 0.01 M HCl) was transferred to an Eppendorf tube and the radioactivity was measured.

[0129] 200–300 μL of labeling buffer (0.25 M NH OAc, pH 5.2) was mixed with the isotope, and EtOH was added to the reaction mixture to make 5% by volume.

[0130] Peptides were added from stock solutions to reach the desired molar radioactivity and the reaction volume was adjusted with labeling buffer to a final volume of 500 μL.

[0131] The pH in the reaction mixture was measured with indicator paper and estimated to be 5.0-5.5.

[0132] The reaction mixture was stirred at 600 rpm at 80° C. for 15 minutes and then cooled to room temperature.

[0133] The incorporation of 67Cu into the DOTA-AE105 conjugate was monitored for the crude reaction mixture by radio-TLC and radio-HPLC. The product (67Cu-DOTA-AE105) was purified by SPE on a tC2 SepPak column by quenching the reaction mixture with DTPA (50 mM, pH 7.0, 2 μL) and subsequently diluting with HO to a final volume of 5 mL. The resulting reaction mixture was loaded onto an SPE column preconditioned with absolute EtOH (5 mL) followed by HO (10 mL). After compound loading, the column was washed with HO (5 mL), and the product was eluted from the column in 500 μL fractions with EtOH in HO (1 mL, 50%, v / v). Alternatively, the product can be eluted with 250 μL of absolute EtOH.

[0134] Unlabeled compound was added to the formulated compound and diluted with formulation buffer (saline) to reach a molar specific radioactivity (60 or 30 MBq / nmol) and a radioactivity concentration of 400 or 200 MBq / mL (60 or 30 MBq / 150 μL).

[0135] The final product (EOS) was analyzed by radio-TLC and radio-HPLC.

[0136] After the final injection, the labeled compounds were analyzed by radio-HPLC, followed by stability analysis by radio-HPLC 4 hours after EOS.

[0137] The results are shown in Tables 1, 2 and 3 below.

[0138] [Table 1]

[0139] [Table 2]

[0140] [Table 3]

[0141] 225Ac labeling of DOTA-AE105 conjugate A radionuclide-labeled uPAR-binding peptide conjugate according to the present invention was prepared by 225Ac labeling of a DOTA-AE105 conjugate.

[0142] The compound was labeled with a molar radioactivity of 30 kBq / nmol for ex vivo distribution and 40 kBq / nmol for potency.

[0143] The desired amount of radioactivity from the 225Ac stock solution (in 0.04 M HCl) was transferred to an Eppendorf tube.

[0144] Labeling buffer (1 M ascorbic acid, pH 6.0) was mixed with the isotope in a 1:1 ratio, ensuring that the compound concentration in the reaction mixture was greater than 90 nmol / mL.

[0145] Peptides were added from stock solutions to reach the desired molar radioactivity.

[0146] The pH in the reaction mixture was measured by indicator paper and estimated to be 5.7-6.0.

[0147] The reaction mixture was stirred at 600 rpm at 90°C for 15 min and then cooled to room temperature.

[0148] The incorporation of 225Ac into the DOTA-AE105 conjugate was monitored for crude reaction mixtures by radio-TLC. Upon completion, the TLC plate was cut in half and immediately measured twice by gamma counting. The Ac-225 radioactivity of each half was estimated based on the 221Fr and 213Bi peaks in the spectrum.

[0149] Compounds were formulated for ex vivo in vitro distribution based on compound quantity. For potency, compounds were formulated based on radioactivity. The formulated compound was diluted with formulation buffer (saline) to achieve a compound concentration of 6.67 nmol / mL (1 nmol / 150 μL) for ex vivo in vitro distribution and a radioactivity concentration of 533.33 MBq / mL (80 kBq / 150 μL) for potency.

[0150] The final product (EOS) was analyzed by radio-TLC and radio-HPLC. The TLC plate was cut in half and measured by gamma counting after constant equilibration. Radio-HPLC fractions were collected at 30-second intervals by automatic fraction collection at 1-minute intervals before and after the peak retention time of the lanthanum-labeled compound standard. The fractions were measured by gamma counting after constant equilibration.

[0151] The results are shown in Tables 1b and 1c below and in Figure 12. Figure 12a shows a representation of the HPLC chromatogram of 30 kBq / nmol Ac-225-DOTA-AE105 based on gamma-counted HPLC fractions. Figure 12b shows a representation of the HPLC chromatogram of 40 kBq / nmol Ac-225-DOTA-AE105 based on gamma-counted HPLC fractions.

[0152] [Table 4]

[0153] [Table 5]

[0154] Example 2: Biodistribution of 67Cu-DOTA-AE105 in mice The biodistribution of 67Cu-DOTA-AE105 was evaluated in female NMRI nude mice bearing subcutaneous U87MG tumors. The U87MG tumor cell line is a human glioblastoma cell line.

[0155] Animals were stratified into four groups (A, B, C, and D) based on tumor volume and body weight 2 days before the study. Group A animals were injected with 26.4 ± 0.4 MBq, Group B animals received 55.1 ± 0.2 MBq, Group C animals received 54.3 ± 0.5 MBq, and Group D animals received 55.2 ± 0.2 MBq (mean ± SEM). Biodistribution was assessed using SPECT / CT imaging at 2, 24, 48, 96, and 168 hours postinjection. Furthermore, conventional ex vivo biodistribution was assessed at final imaging time points of 96 and 168 hours.

[0156] The experimental setup is summarized in Table 4 below.

[0157] [Table 6]

[0158] The results are shown in Figure 1 as percent injected dose per gram (%ID / g) and corrected for radioactive decay. The data in Figure 1 were obtained from in vivo SPECT / CT imaging.

[0159] From Figure 1, it can be seen that 67Cu-DOTA-AE105 exhibits very high tumor uptake and good tumor retention for 48 hours. Furthermore, 67Cu-DOTA-AE105 exhibits low uptake in other organs such as the heart, kidney, and muscle, indicating a good safety profile of the drug.

[0160] Figure 2 shows axial, coronal, and maximum intensity projection (MIP) images of one representative animal from Group A (30 MBq) (Figure 2a) and Group B (60 MBq) (Figure 2b), with arrows indicating the location of the tumor. More specifically, Figure 2a shows the tumor at approximately 30 MBq. 67 Representative axial, coronal, and MIP images are shown 2, 24, 48, 96, and 168 hours after injection of Cu-DOTA-AE105. The image on the left is an axial slice across the tumor center, and the middle image is a coronal slice. The image on the right is a MIP image. Figure 2b shows representative axial, coronal, and MIP images 2, 24, 48, 96, and 168 hours after injection of approximately 60 MBq of 67Cu-DOTA-AE105. The image on the left is an axial slice across the tumor center, and the middle image is a coronal slice. The image on the right is a MIP image.

[0161] The tumor binding and retention results are also summarized in Table 5 below, from which it can be seen that 67Cu-DOTA-AE105 shows very high tumor uptake and good tumor retention at 48 hours for both the 30MBq and 60MBq doses, especially for the 30MBq dose which can be observed in Table 5a.

[0162] [Table 7]

[0163] [Table 8]

[0164] Example 3: Binding specificity To evaluate whether coadministration of unlabeled AE105 could block 67Cu-DOTA-AE105 accumulation, a small-scale study with four animals was conducted. Animals were stratified into two groups: a blocking group consisting of three mice and a non-blocking group consisting of one mouse. Three animals in the blocking group received a 1000-fold excess of unlabeled AE105 immediately before injection with 18 MBq of 67Cu-DOTA-AE105. Animals in the non-blocking group were injected with 67Cu-DOTA-AE105 alone. Inhibition of tumor uptake was assessed by in vivo biodistribution (SPECT / CT imaging) 2 hours after injection. The results are shown in Figure 3. The bars in the figure represent the mean ± SEM. Figure 3 demonstrates that injection of a 1000-fold excess of unlabeled AE105 blocked tumor uptake by more than 50%.

[0165] Example 4: Therapeutic efficacy in xenografted mice bearing GBM The therapeutic efficacy of 67Cu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous U87MG tumors. The U87MG tumor cell line is a human glioblastoma cell line.

[0166] Animals were stratified into four groups (A, B, C, and D) based on tumor volume and body weight 2 days before the study. The mean inclusion volume was 131.2±8.6 mm 3 (Range 66.0~258.3mm 3 ) and the inclusion weight was 29.5 ± 0.4 g (range 24.2–32.9 g). No significant differences were observed between groups in tumor volume or animal weight (one-way ANOVA, p > 0.9 and p = 0.3, respectively).

[0167] Animals were administered either vehicle (Group A) or 67Cu-DOTA-AE105 (Groups B, C, and D) at different radioactivity levels on study day 0. Group B animals received 29.6 ± 0.1 MBq, Group C animals received 29.6 ± 0.1 MBq, and Group D animals received 59.5 ± 0.2 MBq. On study day 14, Group C animals received a booster dose of 30.1 ± 0.1 MBq. Data are presented as measured dose minus residual dose. No decay correction was applied.

[0168] After the first dose, tumor volumes and body weights were monitored twice weekly. Efficacy was evaluated over a 35-day period. The mean tumor volumes of animals included in the study are shown in Figure 4. Differences in tumor volume were assessed at the latest time point at which at least 50% of animals were alive within each compared group.

[0169] On study day 17, all treatment groups had significantly smaller tumor volumes compared to the vehicle control group (Dunnett's test; A vs. B: p<0.03, A vs. C: p=0.02, A vs. D: p<0.0001).

[0170] Animals were euthanized according to a humane endpoint, which included a tumor size of 1500 mm 3 Kaplan-Meier curves of the survival data are shown in Figure 5. The median survival time for animals in vehicle control group A was 17 days, while the median survival times for animals in groups B, C, and D were 21, 21, and 29 days, respectively.

[0171] Body weight did not differ between groups during the study, indicating that the drug was safe and well tolerated.

[0172] Example 5: Comparison of 67Cu-DOTA-AE105 and 177Lu-DOTA-AE105 The tumor binding and retention and treatment efficacy of 67Cu-DOTA-AE105 were compared with 177Lu-DOTA-AE105 in a glioblastoma mouse model.

[0173] Tumor binding and retention of 177Lu-DOTA-AE105 was evaluated in female NMRI nude mice bearing subcutaneous U87MG tumors.

[0174] On study day 0, animals were administered approximately 50 MBq of 177Lu-DOTA-AE105.

[0175] Tumor binding and retention of 177Lu-DOTA-AE105 was assessed by SPECT / CT imaging at 0.5 hours, 2 hours, and 24 hours after injection. The tumor binding and retention results for 177Lu-DOTA-AE105 and the data for 67Cu-DOTA-AE105 in Example 2 (Table 5b) are summarized in Table 6 below.

[0176] [Table 9]

[0177] [Table 10]

[0178] From Table 6a, it can be seen that 177Lu-DOTA-AE105 initially binds to subcutaneous tumors at a level of 0.749% injected dose per gram of body weight (%ID / g) but is rapidly cleared, indicating limited tumor binding and poor tumor retention.

[0179] From Table 6b, it can be seen that 67Cu-DOTA-AE105 exhibits very high tumor uptake of 6.61%ID / g and good tumor retention for 48 hours.

[0180] Furthermore, the efficacy of 177Lu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous U87MG tumors.

[0181] Animals were stratified into three groups (A, B, C) based on tumor volume and body weight on study day 0. The mean inclusion volume was 135.8 ± 814.6 mm 3(Range 15.0~339.0mm 3 ) and the inclusion weight was 23.6 ± 0.4 g (range 29.4–37.4 g). No significant differences were observed between groups in tumor volume or animal weight (one-way ANOVA, p = 0.4).

[0182] Animals were administered either vehicle (Group A) or 177Lu-DOTA-AE105 (Groups B and C) at different radioactivity levels on study day 0. Group B animals received 92.1 ± 1.7 MBq, and Group C animals received 32.9 ± 0.4 MBq. On study day 4, Group C animals received an additional 26.7 ± 0.2 MBq, and on study day 8, Group C animals received 30.7 ± 0.2 MBq. Dose is calculated by subtracting the residual dose from the radioactivity measured before injection. No decay correction was applied.

[0183] After the first dose, tumor volume and body weight were monitored twice weekly. Efficacy was evaluated for 5 weeks after the first dose. Each treatment group was compared to the vehicle control group A.

[0184] The mean tumor volumes of the animals included in the study are shown in Figure 6. Differences in tumor volume were assessed at the latest time point at which at least 50% of the animals were alive within each compared group.

[0185] Comparison of tumor volumes between the vehicle control and treatment groups on study day 18 showed no significant differences between groups (one-way ANOVA, Dunnett's test, A vs. B; p=0.32 and A vs. C; p=0.84).

[0186] Example 6: Therapeutic effects of 67Cu-DOTA-AE105 in glioblastoma and non-small cell lung cancer The therapeutic efficacy of Cu-DOTA-AE105 was tested in female NMRI nude mice bearing either subcutaneous U87MG tumors (see Figure 7a) or NCI-H1993 tumors (see Figure 7b). The U87MG tumor cell line is a human glioblastoma cell line, and the NCI-H1993 tumor cell line is a human non-small cell lung cancer (NSCLC) cell line.

[0187] In each experiment, animals were stratified into two groups based on tumor volume and body weight 2 days before the test. No significant differences were observed between the groups in tumor volume or animal weight.

[0188] Animals were either left untreated or administered 60 Mbq of 67Cu-DOTA-AE105 on day 0 of the study.

[0189] After drug administration, tumor volume and body weight were monitored twice weekly. Efficacy was evaluated over a 20-day (glioblastoma study) or 25-day (non-small cell lung cancer study) period. The mean tumor volume of animals included in the non-small cell lung cancer study is shown in Figure 7b. Differences in tumor volume were assessed at the latest time point at which at least 50% of animals were alive within each compared group.

[0190] All treatment groups had significantly smaller tumor volumes compared to the vehicle control group.

[0191] Animals were euthanized according to a humane endpoint, which included a tumor size of 1500 mm 3 It was reached when it exceeded the limit.

[0192] Body weight did not differ between groups during the study, indicating that the drug was safe and well tolerated.

[0193] In Figure 7, it can be observed that a single administration of 60 Mbq of 67Cu-DOTA-AE105 on day 0 clearly inhibited tumor growth in both glioblastoma-bearing xenografted mice (Figure 7a) and NSCLC-bearing xenografted mice (Figure 7b, showing the mean tumor volume) throughout the study period.

[0194] Example 7: Therapeutic effect of 67Cu-DOTA-AE105 in colorectal cancer The therapeutic efficacy of 67Cu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous HT29 tumors. The HT29 tumor cell line is a human colorectal tumor cell line.

[0195] Animals were stratified into three groups based on tumor volume and body weight two days before the study. No significant differences in tumor volume or animal weight were observed between the groups. Group 1 was left untreated, Group 2 was treated with 60 MBq on study day 0, and Group 3 was treated with 60 MBq on study day 0 and again on study day 7.

[0196] Xenograft tumors were harvested on days 0 (baseline), 2, and 10 after treatment and subjected to analysis by flow cytometry and immunohistochemistry.

[0197] Flow cytometry was performed using standard methods known to those skilled in the art. uPAR-positive cells were labeled using fluorescent uPAR antibodies, and the number of viable uPAR-positive cells in xenograft tumors was measured. The results are shown in Figure 8. Figure 8a shows the percentage of viable cells that were uPAR-positive. Figure 8b shows the percentage of viable cancer cells that were uPAR-positive. Figure 8c shows the mean fluorescence intensity (MFI) of the uPAR marker (fluorescent antibody) for viable cells. Figure 8d shows the MFI of the uPAR marker relative to cancer cells. Figure 8e shows the cutoff applied to define uPAR-positive cells in flow cytometry analysis.

[0198] Immunohistochemistry was performed using standard methods on sliced ​​tumor tissue samples, which were stained for DNA double-strand breaks (using a yH2AX-specific antibody), apoptosis (using cCASP-3, c and p-specific antibodies), and uPAR (using a uPAR-specific antibody).

[0199] From Figure 8, it can be seen that uPAR-positive cancer cells are reduced over 10 days after treatment with 60 MBq. Furthermore, it can be seen that progressive killing of uPAR-positive cancer cells is observed over time, with a further increase after two doses of the test drug.

[0200] From the immunohistochemistry in Figure 9, it can be seen that treatment with 67Cu-DOTA-AE105 induces apoptosis as well as DNA double-strand breaks in human colorectal cancer.

[0201] Example 8: Biodistribution of 225Ac-DOTA-AE105 The biodistribution of 225Ac-DOTA-AE105 was evaluated in female NMRI nude mice bearing subcutaneous U87MG tumors. The U87MG tumor cell line is a human glioblastoma cell line.

[0202] Animals were stratified into two groups (A and B) based on tumor volume and body weight 2 days before the study. Animals in group A were injected with 30 kBq of 225Ac-DOTA-AE105 with a specific activity of 30 kBq / nmol, while animals in group B received a 1000x dose of DOTA-AE105 (non-radioactive construct) immediately before receiving 30 kBq of 225Ac-DOTA-AE105 with a specific activity of 30 kBq / nmol. Biodistribution was assessed using conventional ex vivo biodistribution analysis 1 hour (1H) after injection. Results are shown in Figure 10 as percent of injected dose per gram (%ID / g) and corrected for radioactive decay.

[0203] From Figure 10, it can be seen that the biodistribution of the alpha-emitting uPAR-targeted therapeutic compound 225Ac-DOTA-AE105 shows high uptake (3% ID / g) in human xenograft glioblastoma tumors (U87MG). The inhibition by non-radioactive DOTA-AE105 demonstrates the specificity of tumor binding of 225Ac-DOTA-AE105, as explained by the much lower uptake in group B tumors.

[0204] Example 9: Tolerability and safety of 225AC-DOTA-AE105 The tolerability and safety of 225-AC-AE105 was evaluated in female NMRI nude mice. Animals were stratified into two groups based on body weight 2 days before the study.

[0205] Control animals received vehicle (negative control) on study day 0, and treatment animals received 80 kBq of 225Ac-DOTA-AE105 on study day 0. Animal weights were monitored over 25 days and the results are shown in Figure 11.

[0206] FIG. 11 shows that the animal weights remained stable throughout the study, indicating that the alpha-emitting uPAR-targeted therapy administered at a dose of 80 kBq was well tolerated.

[0207] Example 10: Therapeutic effect of 225Ac-DOTA-AE105 on glioblastoma The efficacy of 225Ac-DOTA-AE105 was evaluated in mice bearing glioblastoma xenograft tumors (U87.MG). Mice were administered a single dose of 30 KBq of 225Ac-DOTA-AE105, and ex vivo apoptosis analysis was performed on day 10 by immunohistochemical staining for caspase-3, a marker of apoptosis. The results are shown in Figure 13 and clearly demonstrate that apoptosis was induced in the active group (the group treated with 225Ac-DOTA-AE105), but not in the vehicle (control) group.

[0208] Example 11: uPAR binding properties of 212Pb-DOTA-AE105 The binding properties of lead (Pb)-labeled DOTA-AE105 to its target, i.e., uPAR, were characterized by surface plasmon resonance (SPR) to determine the binding constant (K), a measure of the affinity of the conjugate with its target. D )

[0209] Characterizing the interaction between a target protein (in this case uPAR) and a ligand by SPR is a method well known to those skilled in the art.

[0210] The target protein, uPAR, was captured on a sensor chip via its protein tag. Test compounds (Pb2+-DOTA-AE105 and Cu2+-DOTA-AE105, respectively) were dissolved in running buffer (10 mM HEPES buffer, pH 7.4, 150 mM NaCl, 50 mM EDTA, 0.05% Tween-20) and prepared in a 5-point, 3-fold dilution series with the highest test compound concentration of approximately 500 nM. Assays were performed at 18°C.

[0211] The results are shown in Table 7 below. From the table below, the cold version of 212Pb-DOTA-AE105 had a measured binding constant (K D It can be seen that the affinity of Pb2+-DOTA-AE105 was 17.4 nM, whereas that of the corresponding cold version of 67Cu-DOTA-AE105 was 182.0 nM. Therefore, the affinity of Pb2+-DOTA-AE105 was more than 10 times higher, demonstrating that even better therapeutic efficacy can be expected with 212Pb-DOTA-AE105. D The difference is mainly due to the very slow dissociation rate of Pb2+-DOTA-AE105, which is necessary for the radionuclide to remain on target and be effective, for example, in therapeutic applications.

[0212] [Table 11]

Claims

1. A radionuclide-labeled uPAR-binding peptide conjugate comprising: a) uPAR-binding peptide b) a chelating agent suitable for binding to radioactive metals c) a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb, and Tb Including, A radionuclide-labeled uPAR-binding peptide conjugate, wherein said peptide is bound to said radionuclide by said chelating agent.

2. a) uPAR-binding peptide b) a chelating agent suitable for binding to radioactive metals c) a radionuclide selected from the group consisting of Cu, Ac, Pb, Tb, and Tb It consists of 2. The radionuclide-labeled peptide conjugate of claim 1, wherein the peptide is bound to the radionuclide by the chelating agent.

3. The uPAR-binding peptide is a monomeric or multimeric peptide, and the one or more peptide components are: (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Ser)-(Leu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Gln)-(Tyr)(Leu)-(Trp)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(Ser)-(D-Arg)-(Tyr)-Leu)-(Trp)-(Ser), (D-Thr)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-([β]-2-naphthyl-L-alanine)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Arg)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(Ser), (D-Glu)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(Tyr)-(Tyr)-(Leu)-(Trp)-(Ser), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Leu)-(Leu)-(Trp)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-([β]-cyclohexyl-L-alanine)-(Leu)-(Trp)-(Ile), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)([β]-1-naphthyl-L-alanine)-(D-His), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(3-indolylethyl)glycine)-(N-(2-methoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-benzylglycine)-(N-(2[β]thoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthalyl)glycine)-(N-(2-methoxyethyl)glycine), and (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(2,3-dimethoxybenzyl)glycine)-(Ile) 3. The radionuclide-labeled peptide conjugate of claim 1, selected from the group consisting of:

4. 4. The radionuclide-labeled peptide conjugate of any one of claims 1 to 3, wherein the uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser).

5. 5. The radionuclide-labeled uPAR-binding peptide conjugate of any one of claims 1 to 4, wherein the chelator is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, sarcofagin, CB-TE2A, or derivatives thereof.

6. 6. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, and sarcofazine.

7. 7. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, wherein the chelating agent is DOTA.

8. 8. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, wherein the radionuclide is 67Cu, 225Ac, or 212Pb.

9. 9. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, wherein the radionuclide is 67Cu.

10. formula: 【Chemistry 1】 10. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, having the formula:

11. 9. The radionuclide-labeled uPAR-binding peptide conjugate of claim 1, wherein said radionuclide is 67Cu and said chelating agent is DOTA.

12. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of claims 1 to 11 for use as a pharmaceutical.

13. A radionuclide-labeled uPAR-binding peptide conjugate according to any one of claims 1 to 11 for use in the treatment of uPAR-expressing cancer.

14. 14. The radionuclide-labeled uPAR-binding peptide conjugate according to any one of claims 1 to 11 for use according to claim 13, wherein the cancer is selected from the group consisting of breast cancer, brain cancer, in particular high-grade glioma, glioblastoma, NSCLC (non-small cell lung cancer), SCLC (small cell lung cancer), neuroendocrine tumors, head and neck cancer, HNSCC (head and neck squamous cell carcinoma), pancreatic cancer, prostate cancer, CRC (colorectal cancer), and gastric cancer.

15. 15. The radionuclide-labeled uPAR-binding peptide conjugate of any one of claims 1 to 11 for use according to claim 13 or claim 14, wherein the cancer is selected from the group consisting of breast cancer, NSCLC, neuroendocrine tumors, HNSCC, pancreatic cancer, prostate cancer, CRC, glioblastoma and gastric cancer.

16. The radionuclide-labeled uPAR-binding peptide conjugate according to any one of claims 1 to 11 for use according to any one of claims 13 to 15, wherein the cancer is selected from the group consisting of glioblastoma.

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