Urokinase plasminogen activator receptor (UPAR)-PET / CT in brain tumors

Abstract

The present invention relates to a positron emission imaging agent for the prognostic assessment of a brain tumor in a patient by PET imaging of cancer wherein the imaging agent comprises a uPAR binding peptide coupled to a radionuclide. The invention also relates to a composition comprising a radiopharmaceutical for treating or alleviating a brain tumor in a subject, wherein the radiopharmaceutical comprises a radionuclide and a uPAR binding peptide.

Description

Technical Field

[0001] This invention relates to a positron emission tomography (PET) imaging agent for prognostic assessment of brain cancer patients via cancer PET imaging, wherein the imaging agent comprises a radionuclide bound by a chelating agent NOTA or DOTA. 68 Ga or 64 Cu-coupled uPAR-binding peptide. The invention also relates to compositions comprising radiopharmaceuticals for treating or alleviating brain cancer in subjects, wherein the radiopharmaceutical comprises a radionuclide and a uPAR-binding peptide. Finally, the invention relates to the use of the imaging agent as a companion diagnostic reagent to therapeutic radiopharmaceuticals. Background Technology

[0002] Gliomas are one of the most common types of brain cancer, with an annual incidence of 6 cases per 100,000 people (1). These highly heterogeneous tumors are classified into four distinct WHO grades using a multi-level grading system. Grade 1–2 gliomas are referred to as low-grade gliomas (LGG), while grade 3–4 tumors are referred to as high-grade gliomas (HGG). The increase in WHO grade is associated with increased tumor invasiveness and poorer survival (2–4). Despite significant advancements in oncology, survival rates for glioma patients have remained largely unchanged, with a 5-year survival rate of 82% for LGG and a very poor survival prospect of only 3–10% for HGG (4–6). Treatment for gliomas varies considerably depending on the tumor subtype. For LGG, treatment options range from postoperative observation (biopsy, partial or total resection) to radiotherapy alone or in combination with chemotherapy, including procarbazine, lomustine, vincristine regimens (PCV), or temozolomide (TMZ). For HGG, the goal of treatment is complete tumor resection, followed by combined radiotherapy and chemotherapy with TMZ or PCV (3). This variability in treatment regimens underscores the need for phenotyping and risk stratification of gliomas before the onset of treatment to ensure more precise management of these tumors.

[0003] Magnetic resonance imaging (MRI) is the standard imaging modality for detecting gliomas and can be supplemented by positron emission tomography (PET), with amino acid tracers such as O-(2-[18F]fluoroethyl)-1-tyrosine (FET) being particularly recommended (7-9). FET-PET has a variety of applications, including diagnosis, prognostic assessment, target delineation, and determination of tumor recurrence (9).

[0004] In addition, tracers using the targeted proteolytic urokinase plasminogen activator (uPA) system [ 68 Ga]Ga-NOTA-Asp-Cha-Phe-D-Ser-D-Arg-Tyr-Leu-Trp-Ser-OH ( 68PET imaging of Ga-NOTA-AE105 is emerging as a promising new imaging biomarker for diagnosis, prognostic assessment, risk stratification, and therapeutic targets for solid cancers. Over the years, several studies have demonstrated the applicability of the uPA receptor (uPAR) as a diagnostic biomarker for cancers associated with poor disease prognosis (10). uPAR is highly upregulated in most solid cancers and expressed in limited quantities in normal tissues. It is located on the cell surface, where it binds to the serine protease uPA. This promotes cell proliferation, angiogenesis, proteolysis, and motility, leading to tumor progression and invasion of surrounding tissues (10-12).

[0005] To target uPAR, a PET radiotracer in which the targeting peptide is a high-affinity antagonist of uPAR was developed. 68 Ga-NOTA-AE105 and 64 Cu-DOTA-AE105 (13-15). In two phase 1 trials involving primary and metastatic tumors, 68 Ga-NOTA-AE105 and 64 The safety, biodistribution, and radioligand accumulation of Cu-DOTA-AE105 in cancerous tissues have been established. Immunohistochemistry confirmed histopathological accumulation of the tracer in cancerous tissues consistent with uPAR expression (13, 36). Furthermore, 68 The utility of Ga-NOTA-AE105 for uPAR-PET as a promising non-invasive assessment method for localized prostate cancer has been demonstrated, with high diagnostic accuracy in distinguishing between low-risk and intermediate-risk Gleason score distributions (16). uPAR-PET has also been found to be highly prognostic in neuroendocrine tumors (17) and head and neck cancers (18). In gliomas, uPAR-PET has been highlighted as an effective imaging biomarker for tumor visualization using human orthotopic xenograft models of glioblastoma (19).

[0006] From a therapeutic perspective, uPAR has been identified as a promising target for peptide receptor radionuclide therapy (PRRT), and the efficacy of uPAR-targeted PRRT has been previously demonstrated in preclinical models of prostate and colorectal cancer (20, 21). Furthermore, recent work has revealed a strong correlation between uPAR expression on uPAR-PET and overall survival (OS) and progression-free survival (PFS) in patients with neuroendocrine tumors, highlighting uPAR as a promising target for PRRT therapy. In fact, 68% of patients across all tumor grades are uPAR-positive (17).

[0007] Therefore, improved prognostic tools related to brain tumors would be beneficial, especially more effective and / or more reliable treatments for brain tumor subtypes. Summary of the Invention

[0008] uPAR-PET has been found to serve as a prognostic marker for tumor invasiveness in brain cancers such as gliomas, and uPAR-PET-positive brain cancers may be a future candidate for peptide receptor radionuclide therapy (PRRT) targeting uPAR.

[0009] Therefore, the aim of this PET / MRI study of uPAR-targeted PRRT (using 68Ga-NOTA-AE105 as an example) in patients with primary gliomas was to investigate the relationship between uPAR-targeted PRRT uptake and overall survival (OS) and progression-free survival (PFS). Furthermore, the aim was to determine the proportion of uPAR-PET-positive tumors to assess how many of these patients might be eligible for future uPAR-PRRT.

[0010] Therefore, the object of the present invention relates to providing improved prognostic tools for brain tumors.

[0011] Specifically, the present invention also aims to provide improved treatment options for patients with brain tumors.

[0012] Therefore, one aspect of the present invention relates to a positron emission tomography (PET) imaging agent for prognostic assessment of progression-free survival (PFS) and / or overall survival (OS) of brain tumors in patients via cancer PET imaging. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 maximum and / or average SUV levels above the threshold indicate poor prognosis in progression-free survival (PFS) and / or overall survival (OS); and The maximum SUV and / or the average SUV (quantitative) level, which are equal to or below the threshold level, indicate a good prognosis for progression-free survival (PFS) and / or overall survival (OS).

[0013] Two uPAR-PET tracers 64 Cu-DOTA-AE105 and 68 Ga-NOTA-AE105 shares the same binding moiety, the peptide AE105. Furthermore, both have been tested in humans and animals (see, for example, WO2014 / 086364 A1) and have shown similar uptake in breast, bladder, and prostate cancers. Therefore, not limited to theory, it is believed that PET tracers containing the following will also act in a similar manner for patients with brain tumors: -As a radioactive nuclide 64 Cu or 68 Ga; - DOTA or Nota as chelating agents; and - The uPAR-binding peptide according to the present invention.

[0014] Preferably, the developing agent comprises a uPAR-binding peptide coupled to the radionuclide 68Ga via the chelating agent NOA.

[0015] As described above, the method of the present invention and the composition used according to the present invention can also be used as companion diagnostic reagents.

[0016] Therefore, another aspect of the present invention relates to a positron emission tomography (PET) imaging agent used as a companion diagnostic agent for patients with brain tumors via cancer PET imaging. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; in The maximum and / or average SUV levels above a threshold level indicate that uPAR-binding drugs, such as uPAR-binding radiopharmaceuticals, will be effective against the brain tumor; and The maximum and / or average SUV levels, which are equal to or below the threshold level, indicate that uPAR drugs, such as uPAR combined with radiopharmaceuticals, will be ineffective against the brain tumor. The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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).

[0017] Another aspect of the invention relates to a composition comprising a radiopharmaceutical for treating or alleviating a brain tumor in a subject; The radiopharmaceutical contains a radionuclide and a uPAR-binding peptide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or a uPAR-binding variant thereof; and The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 subjects in this study have been diagnosed with brain tumors that express uPAR, and uPAR expression is above a predetermined threshold level. The threshold level is determined using a positron emission tomography (PET) imaging agent as defined according to the present invention. Attached Figure Description

[0018] Figure 1 Figure 1 The flowchart of the inclusion process for the Common Standards for Clinical Reporting Trials (CONSORT) is shown.

[0019] Figure 2 Figure 2 Examples of uPAR PET / MRI performed in a patient with temporoparietal IDH wild-type, WHO grade 4 (MGMT unmethylated) glioblastoma (tumor SUV maximum value 3.3) are shown. A) Gadolinium-contrast T1W MPRAGE MRI B) T2W FLAIR MRI image C) uPAR-PET image D) T2W FLAIR MRI and uPAR-PET fusion image. Color scale range: 0 to tumor SUV maximum value 3.3.

[0020] Figure 3 Figure 3 This image shows an example of uPAR PET / MRI performed on a patient with IDH wild-type, WHO grade 4 (MGMT unmethylated) glioblastoma involving the genu of the corpus callosum (tumor SUV maximum value 2.2). A) Gadolinium-contrast T1W MRI B) T2W FLAIR MRI image C) UPAR-PET image D) T2W FLAIR MRI and uPAR-PET fusion image. Color scale range: 0 to tumor SUV maximum value 2.2.

[0021] Figure 4 Figure 4 shows the Kaplan-Meil survival curves for primary gliomas: (A) overall survival (OS) curves grouped by the maximum SUV value of 1.1, and (B) progression-free survival (PFS) curves grouped by the maximum SUV value of 0.635.

[0022] Figure 5 Figure 5 shows the Kaplan-Mel survival curves for primary HGG, (A) overall survival (OS) curves grouped by SUV maximum value 1.1, and (B) progression-free survival (PFS) curves grouped by SUV maximum value 1.1.

[0023] The invention will now be described in more detail below. Detailed Implementation

[0024] definition Before discussing the invention in further detail, the following terms and conventions will be defined first: brain tumor Brain tumors occur when abnormal cells form in the brain. Gliomas are one of the most common types of brain cancer. Gliomas can be classified as high-grade gliomas (HGG) (WHO grades 3 and 4) and low-grade gliomas (LGG) (WHO grades 1+2). WHO grade 4, also known as glioblastoma, is the predominant component (approximately 80%) of high-grade gliomas.

[0025] 68 Ga Gallium-68.

[0026] 64 Cu Copper-64.

[0027] 177 Lu 177 Lu is a low-energy β-emitter (with a maximum penetration of ~1.5 mm in soft tissue) that induces cytotoxic effects in tumors through "crossfire" and "side-by" effects via direct β particles and Auger electrons, respectively, but exhibits little or no toxicity in surrounding tissues. Therefore, 177 Lu is considered the best radionuclide for treating tumors of all sizes and / or disseminated metastatic diseases.

[0028] AE105 Ac-Asp-Cha-Phe-(D)Ser-(D)Arg-Tyr-Leu-Trp-Ser The peptides described in this invention can be synthesized, for example, by standard solid-phase peptide chemistry.

[0029] NOTA NOTA: 2,2',2"-(1,4,7-triazacyclononane-1,4,7-triyl)triacetic acid.

[0030] NOTA can bind to AE105, thus providing NOTA-AE105 (NOTA-Asp-Cha-Phe-Ser-Arg-Tyr-(Leu-Trp-Ser)). This can be illustrated by the following chemical structure: In one implementation, the developer is 68 Ga-NOTA-AE105.

[0031] .

[0032] DOTA DOTA (also known as tetraxetan) is an organic compound with the molecular formula (CH2CH2NCH2CO2H)4. The molecule consists of a central 12-membered tetraaza ring (i.e., containing four nitrogen atoms). DOTA is used as a complexing agent, particularly for lanthanide ions. Its complexes are used in medicine as contrast agents and cancer treatments.

[0033] DOTA's preferred IUPAC name 2,2',2'',2'''-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid Below is an example of DOTA and 68 Ga and 64 CU network merging with AE105.

[0034] Progression-free survival (PFS) The term "progression-free survival" (PFS) is defined as "the length of time a patient has a disease but does not worsen or progress during or after treatment or intervention for a disease such as cancer".

[0035] Relapse-free survival (RFS) The term “recurrence-free survival” (RFS) is defined as a clinical endpoint, which is defined as the time from diagnosis to any recurrence of the disease in a local area (TN site) and / or distant metastasis (M site), wherein death due to other causes is recorded as censored. Disease-free survival (DFS) is defined the same as RFS, but includes death as an event.

[0036] Total Survival The term "overall survival" (OS) is defined as the time from diagnosis to death from any cause.

[0037] Threshold level In the context of this invention, the terms "threshold level," "reference level," or "cut-off" refer to a standard relating to a quantity that can be compared with other values ​​or characteristics.

[0038] In one embodiment of the invention, a threshold level can be determined by studying the uPAR levels of healthy subjects using PET / CT. One or more threshold levels can be calculated by applying various statistical methods, such as critical value finding and multivariate analysis.

[0039] See also Example 5, where a critical value was determined.

[0040] Based on these results, a critical value can be obtained that shows the relationship between the detected level and patients at risk. Therefore, the critical value can be used, for example, to determine uPAR levels that correspond to an increased risk of poor PFS or OS.

[0041] risk assessment The inventors have successfully developed a novel method for predicting the prognosis (e.g., PFS and / or RFS and / or OS) of subjects with brain cancer. To determine whether the risk of a poor prognosis is increased, a cutoff value (reference level) must be established. This cutoff value can be determined by a laboratory, a physician, or based on the specific circumstances of each patient.

[0042] Critical value levels can be established using various methods, including: multivariate statistical tests (such as partial least squares discriminant analysis (PLS-DA), random forest, support vector machine, etc.), percentiles, mean and standard deviation; median; fold change.

[0043] Multivariate discriminant analysis and other risk assessments can be performed on free or commercially available computer statistical software packages (SAS, SPSS, Matlab, R, etc.) or other statistical software packages or screening software known to those skilled in the art.

[0044] It will be apparent to those skilled in the art that, in any of the above embodiments, changing the risk threshold level can alter the results of the discriminant analysis for each subject.

[0045] Statistics can assess the importance of each level. Common statistical tests applied to datasets include t-tests, F-tests, or more advanced tests and data comparison methods. These tests or methods can be used to determine whether two or more samples are significantly different.

[0046] Significance can be determined using standard statistical methods known to those skilled in the art.

[0047] The selected reference level can be changed depending on the mammal / subject being tested.

[0048] Preferably, the subject described in this invention is a human subject.

[0049] As is known in the art, the chosen reference level can be varied if different specificities or sensitivity are required. Sensitivity and specificity are widely used statistics to describe and quantify how good and reliable a biomarker or diagnostic test is. Sensitivity assesses how well a biomarker or diagnostic test detects a disease, while specificity assesses the likelihood that an individual (i.e., a control, a disease-free patient) can be correctly identified as risk-free.

[0050] Several terms are used to describe sensitivity and specificity: true positive (TP), true negative (TN), false negative (FN), and false positive (FP). A diagnostic test result is considered TP if the disease is proven to be present in the affected individual. A test result is TN if the disease is absent in the individual (i.e., the control, the disease-free individual) and the diagnostic test confirms the absence of the disease. A test result is FP if the diagnostic test indicates the presence of the disease in individuals who do not have it. Finally, a test result is FN if the diagnostic test indicates the absence of the disease in the affected individual.

[0051] Sensitivity As used in this article, sensitivity is a measure of the proportion of actual positive cases that are correctly identified, i.e., the percentage of subjects with a higher-than-normal risk of poor prognosis who are identified as having a higher-than-normal poor prognosis.

[0052] Typically, the sensitivity of a test can be described as the proportion of true positives out of the total number of patients with the target disease (i.e., a higher-than-normal risk of poor prognosis). The total number of patients with the target disease is the sum of (detected) true positives (TPs) and (undetected) false negatives (FNs).

[0053] Specificity As used herein, specificity is a measure of the proportion of negative results that are correctly identified; that is, the percentage of mammals without an increased risk of adverse prognosis identified as not having a risk higher than normal. An ideal diagnostic test is one with 100% specificity, meaning it detects only subjects with a risk of adverse prognosis higher than normal, thus eliminating false positives, and one with 100% sensitivity, meaning it detects all subjects with a risk of adverse prognosis higher than normal, thus eliminating false negatives.

[0054] For any test, there is usually a trade-off between each metric. For example, in a manufacturing scenario testing for defects, it might be worthwhile to risk discarding functional parts (low specificity) in order to increase the chance of identifying almost all defective parts. This trade-off can be represented graphically by an ROC curve.

[0055] The chosen specificity determines the percentage of acceptable false positives in a given study / population and at a given institution. Sensitivity can be increased by reducing specificity.

[0056] Those skilled in the art generally understand that screening methods used for prognosis are a decision-making process, and therefore the specificity and sensitivity chosen depend on what the given institution / clinical staff considers the best outcome.

[0057] SUV maximum value In this paper, the term "SUVmax" refers to the "maximum standardized uptake value" (SUVmax), which is widely used to measure the uptake of uPAR and FDG in malignant tissues. Increased uptake values ​​reflect increased uptake or binding of tracers to cancer cells and can be measured using PET imaging.

[0058] SUV average In this article, the term "SUV mean" refers to the average standardized intake value.

[0059] Companion diagnostic reagents In this article, "companion diagnostic reagent" refers to a diagnostic test used as a companion to a therapeutic drug to determine its suitability for a particular person.

[0060] Companion diagnostics can be co-developed with drugs to help select or exclude patient groups treated with a particular drug based on identifying biological characteristics that indicate a response to or non-response to treatment. Companion diagnostics are developed based on accompanying biomarkers, which hold the promise of helping to predict potential reactions or serious toxicities.

[0061] Imaging agents used for prognostic assessment of brain cancer patients via PET scans. Improving prognostic assessment for brain cancer patients is important, for example, for determining the need for radiotherapy and for determining the appropriate radiotherapy. Therefore, improved companion diagnostic agents are also important. Accordingly, one aspect of the present invention relates to a positron emission tomography (PET) imaging agent for the prognostic assessment of progression-free survival (PFS) and / or overall survival (OS) in brain tumor patients via PET imaging of the cancer. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 maximum and / or average SUV levels above the threshold indicate poor prognosis in progression-free survival (PFS) and / or overall survival (OS); and The maximum SUV value and / or the average SUV value and / or the level that are equal to or below the threshold level indicate a good prognosis for progression-free survival (PFS) and / or overall survival (OS).

[0062] As described in Example 5, the imaging agent according to the present invention is a strong prognostic biomarker for patients with brain cancer. Furthermore, an optimal cutoff value has been determined. Again, it is worth noting that such a biomarker is also relevant to companion diagnostic reagents, as it can be used to identify patients who may be susceptible to radiotherapy using radionuclides conjugated to uPAR-binding peptides.

[0063] Different uPAR-binding variants of the uPAR-binding peptide (including AE105) are disclosed in WO2014 / 086364 A1.

[0064] The uPAR binding portion of the developer is preferably referred to as AE105. Thus, in one embodiment, the peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser).

[0065] In one implementation, the radionuclide is selected from... 68 Ga and 64 The group consisting of Cu.

[0066] In another embodiment, the radionuclide is coupled to the uPAR-binding peptide via a chelating agent such as DOTA or NOA.

[0067] In a preferred embodiment, the radionuclide is coupled to the uPAR-binding peptide via the chelating agent NOA. 68 Ga.

[0068] In another preferred embodiment, the developer used has the following formula: .

[0069] In another embodiment, the radionuclide is coupled to the uPAR-binding peptide via a chelating agent, DOTA or NOA. 64 Cu.

[0070] In a preferred embodiment, the developer used has the following formula: .

[0071] In another preferred embodiment, the developer used has the following formula: .

[0072] In one implementation, the imaging agent is administered at a dose of 10-500 MBq, followed by a PET scan 10 minutes to 24 hours after the imaging agent administration, and quantified by the maximum SUV value and / or the average SUV value.

[0073] Brain tumors occur when abnormal cells form in the brain. Gliomas are one of the most common types of brain cancer. Gliomas can be classified as high-grade gliomas (HGG) (WHO grades 3 and 4) and low-grade gliomas (LGG) (WHO grades 1+2). WHO grade 4, also known as glioblastoma, is the predominant component (approximately 80%) of high-grade gliomas.

[0074] In one embodiment, the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma WHO grade 4 (also known as glioblastoma). As shown in Example 3, the method of the present invention can effectively image WHO grade 4 tumors. Furthermore, most cancers that can be imaged are WHO grade 4.

[0075] In another embodiment, the C-terminus is a carboxylic acid or an amide.

[0076] To further improve cancer localization, PET scans can be combined with other scan types. Therefore, in one implementation, prognostic assessment includes PET / CT scans and / or PET / MR scans. In the examples section, PET / CT is used.

[0077] Prognostic assessment can be further defined. Therefore, in one implementation, prognosis is a prediction of progression-free survival (PFS) and / or overall survival (OS). In the Examples section, PFS and OS have been assessed.

[0078] Different amounts (MBq) of developer can be used. Furthermore, the time between developer application and scanning may also vary. Therefore, in one embodiment, the developer is applied at a dose of 10-500 MBq, followed by a PET scan 10 minutes to 24 hours after developer application, and quantified by the maximum SUV and / or the average SUV and / or the TLR.

[0079] In another embodiment, the imaging agent is administered at a dose of 20-400 MBq, such as 50-400 MBq, 70-300 MBq, 100-300 MBq, or 100-300 MBq. Preferably, for example, 150-250 MBq, and more preferably, 170-230 MBq. Thus, in a preferred embodiment, the imaging agent is administered at a dose of 70-300 MBq, preferably, for example, 150-250 MBq. In the examples section, approximately 200 MBq has been used, and PET imaging was performed using a Siemens Biograph 128 mCT. However, other devices may be more sensitive, thus allowing for lower amounts (MBq) of imaging agent.

[0080] In one implementation, a sufficient amount of imaging agent is applied to allow for PET imaging with a radiation dose sufficient for imaging.

[0081] 68Ga has a half-life of about 1 hour (68 min), which provides 10 minutes to 5 hours, preferably 20 minutes to 3 hours, of actual time before a PET scan. 64 Cu has a half-life of approximately 12.7 hours, making it possible to have an interval of 20 minutes to 24 hours before a PET scan.

[0082] Therefore, in one relevant implementation, the PET scan is performed 20 minutes to 10 hours after the application of the imaging agent, for example, 20 minutes to 5 hours after the application of the imaging agent, or for example, 30 minutes to 3 hours after the application of the imaging agent.

[0083] In another embodiment, the imaging agent according to the invention is in a pharmaceutical composition comprising the imaging agent and one or more pharmaceutically acceptable adjuvants, excipients and / or diluents.

[0084] To enable prognostic assessment, threshold levels (critical values / reference levels) may be included. Therefore, in one embodiment, the developer used according to the invention... Maximum and / or average SUV levels above a threshold indicate poor prognosis in progression-free survival (PFS) and / or overall survival (OS); The maximum SUV level and / or the average SUV level, which are equal to or below the threshold level, indicate a good prognosis for progression-free survival (PFS) and / or overall survival (OS).

[0085] As described in the Examples section, the “critical value” / “threshold level” was determined using the Cut-off Finder application (26).

[0086] In Example 5, it has already been... 68 Ga-NOTA-AE105 calculates a specific optimal threshold level (critical value).

[0087] Therefore, in another embodiment, the threshold level for the maximum SUV value and / or the average SUV value is in the range of 0.5-4, such as in the range of 0.5-2, preferably in the range of 0.5-1.5. In Example 5, the optimal threshold levels for PFS and OS were determined.

[0088] In one embodiment, for all gliomas, the maximum and / or average threshold of the SUV associated with PFS is in the range of 0.3-1, preferably 0.4-0.8, more preferably about 0.64.

[0089] In one embodiment, for high-grade gliomas, the maximum SUV and / or average SUV threshold associated with PFS is in the range of 0.7-1.5, preferably 0.9-1.3, and more preferably about 1.1.

[0090] In one embodiment, for all gliomas, the threshold values ​​for the maximum and / or average SUV associated with OS are in the range of 0.7-1.5, preferably 0.9-1.3, and more preferably about 1.1.

[0091] In one embodiment, for all high-grade gliomas, the threshold values ​​for the maximum and / or average SUV associated with OS are in the range of 0.7-1.5, preferably 0.9-1.3, and more preferably about 1.1.

[0092] In a preferred embodiment, the above values ​​are for 68 Ga-NOTA-AE105.

[0093] for 64 Cu-DOTA-AE105 was observed to have 2-4 times higher maximum and / or average SUV values, and the threshold values ​​would also be 2-4 times higher than the values ​​mentioned above.

[0094] Therefore, in another embodiment, the threshold level for the maximum SUV value and / or the average SUV value is in the range of 2-16, such as in the range of 2-8, preferably in the range of 2-6, preferably when using 64 When Cu-DOTA-AE105.

[0095] In one embodiment, for all gliomas, the maximum and / or average threshold of the SUV associated with PFS is in the range of 0.6-4, preferably 0.8-3.2, more preferably about 1.2-2.4, preferably when using 64 When Cu-DOTA-AE105.

[0096] In one embodiment, for high-grade gliomas, the maximum and / or average threshold values ​​of the SUV associated with PFS are in the range of 2.8-24, preferably 3.6-20.8, more preferably about 4.4-17.6, preferably when using 64 When Cu-DOTA-AE105.

[0097] In one implementation, for all gliomas, the maximum and / or average threshold values ​​of the SUV associated with OS are in the range of 1.4-6, preferably 1.8-5.2, more preferably about 2.2-4.4, preferably when using 64 When Cu-DOTA-AE105.

[0098] In one implementation, for all high-grade gliomas, the maximum and / or average threshold values ​​of the SUV associated with OS are in the range of 1.4-6, preferably 1.8-5.2, more preferably about 2.2-4.4, preferably when using 64 When Cu-DOTA-AE105.

[0099] In yet another implementation, the threshold level is determined using a cut-off finding method to obtain the Kaplan-Melto (time series test) split value and the corresponding hazard ratio (HR).

[0100] It should be noted, of course, that embodiments of other aspects of the present invention are also applicable to this aspect.

[0101] A method for in vivo imaging using PET scans to assess the prognosis of brain cancer. Another aspect of the present invention relates to a method for in vivo imaging via PET imaging for assessing the prognosis of a patient with brain cancer, the method comprising: a) Provide subjects who have previously been administered the imaging agent according to the invention; b) Detect the radioactive emission of radioisotopes from the imaging agent applied in step a) by in vivo PET imaging. c) Generate an image representing the location and / or amount of the radioactive emission; d) Determine the distribution and extent of uPAR expression in the subjects, wherein the expression is correlated with the signal emitted by the in vivo imaging agent; and e) Compare the distribution and extent of identified uPAR expression with threshold levels; The maximum and / or average SUV levels above the threshold indicate poor prognosis in progression-free survival (PFS) and / or overall survival (OS); and The maximum SUV level and / or the average SUV level, which are equal to or below the threshold level, indicate a good prognosis for progression-free survival (PFS) and / or overall survival (OS).

[0102] In one implementation, if the indicated progression-free survival (PFS) or overall survival (OS) outcome is favorable, the subject will be scheduled for a less aggressive treatment regimen to avoid unnecessary toxicity. Those skilled in the art will fully understand that a less aggressive treatment regimen differs from the treatment already administered to the relevant patient. Examples include changing the treatment compound (or radiation), reducing the dose, or adjusting the treatment interval.

[0103] In another implementation, the levels of the maximum SUV and the average SUV determine whether a patient is suitable for targeted radionuclide therapy.

[0104] In one aspect, this method forms the basis for companion diagnostic reagents used in targeted radionuclide therapy. Therefore, in such an aspect... Maximum and / or average SUV levels above a threshold level indicate that uPAR-binding drugs, such as uPAR-binding radiopharmaceuticals, will be effective against the brain cancer; and The maximum and / or average SUV levels, which are equal to or below the threshold level, indicate that uPAR-binding drugs, such as uPAR-binding radiopharmaceuticals, will not be effective against the brain cancer.

[0105] Preferably, the drug is a uPAR-bound radiopharmaceutical, and more preferably, the radiopharmaceutical is... 177 Lu-DOTA-AE105 67 Cu-DOTA-AE105 225 Ac-DOTA-AE105 or 212 Pb-DOTA-AE105.

[0106] The use of positron emission tomography (PET) imaging agents in the prognostic assessment of brain cancer in patients via in vivo PET imaging. Another aspect of the invention relates to the use of the positron emission tomography (PET) imaging agent according to the invention in the prognostic assessment of brain cancer patients by in vivo PET imaging of uPAR-expressing tumors. The imaging agent comprises a radionuclide bound by the chelating agent NOA. 68 Ga-coupled uPAR-binding peptide; The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant.

[0107] In an alternative aspect, the developing agent comprises a radionuclide bound by a chelating agent such as NOTA or DOTA. 68 Ga or 64 Cu-coupled uPAR-binding peptide.

[0108] Furthermore, in one implementation, its uPAR binding variant is selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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).

[0109] Companion diagnostic reagents As described above, the method and composition used according to the present invention can also be used as companion diagnostic reagents. Therefore, another aspect of the present invention relates to a positron emission tomography (PET) imaging agent used as a companion diagnostic reagent for brain cancer patients undergoing PET imaging for cancer. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; in The maximum and / or average SUV levels above the threshold indicate that uPAR-binding drugs, such as uPAR-binding radiopharmaceuticals, will be effective against the brain tumor; and The maximum and / or average SUV levels, which are equal to or below the threshold level, indicate that uPAR-bound drugs, such as uPAR-bound radiopharmaceuticals, will be ineffective against the brain tumor. The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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).

[0110] In a preferred embodiment, the developer is 68 Ga-NOTA-AE105. Therefore, in one embodiment, the radionuclide and the chelating agent are... 68 Ga-NOTA. This developer has been used in the examples section.

[0111] In another preferred embodiment, the developer is 64 Cu-DOTA-AE105.

[0112] Therefore, in another embodiment, the radionuclide and the chelating agent are 64 Cu-DOTA.

[0113] In another implementation, the positron emission tomography (PET) imaging agent is a companion diagnostic reagent for the radiopharmaceutical.

[0114] In a preferred embodiment, the positron emission tomography (PET) imaging agent is a companion diagnostic reagent for a radiopharmaceutical as defined in this invention.

[0115] In another preferred embodiment, the positron emission tomography (PET) imaging agent is a companion diagnostic reagent for a radiopharmaceutical as defined according to the present invention, wherein the radionuclide and chelating agent of the radiopharmaceutical are... 177 Lu-DOTA.

[0116] In a preferred embodiment, the positron emission tomography (PET) imaging agent is a companion diagnostic reagent for a radiopharmaceutical having the following formula: .

[0117] As described above, companion diagnostic reagents can be used to determine the suitability of a drug. Therefore, in one embodiment, companion diagnostic reagents are used to determine the suitability of a radiopharmaceutical (such as a radiopharmaceutical as defined according to the present invention) for a particular person.

[0118] In another preferred embodiment, the drug / radiopharmaceutical is a radiopharmaceutical. In an even more preferred embodiment, uPAR combined with a radiopharmaceutical is... 177 Lu-DOTA-AE105 67 Cu-DOTA-AE105 225 Ac-DOTA-AE105 and / or 212 Pb-DOTA-AE105.

[0119] In one embodiment, the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma (WHO grade 4) (glioblastoma).

[0120] In another implementation, the brain tumor is a high-grade glioma (glioblastoma).

[0121] Note again that embodiments of other aspects of the invention are also applicable to this aspect.

[0122] Radiopharmaceuticals used to treat or alleviate brain cancer in subjects. As mentioned above, imaging agents can be used as companion diagnostic reagents. This method can be used to identify brain cancer subtypes that express certain upregulated or downregulated surface proteins. The upregulated surface proteins can be used to target cancer with radiopharmaceuticals.

[0123] Therefore, another aspect of the present invention relates to a composition comprising a radiopharmaceutical for treating or alleviating brain cancer in a subject. The radiopharmaceutical contains a radionuclide and a uPAR-binding peptide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or a uPAR-binding variant thereof; and The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 subjects in this study have been diagnosed with brain tumors that express uPAR, and uPAR expression is above a predetermined threshold level. The threshold level is determined using a positron emission tomography (PET) imaging agent according to the present invention.

[0124] Preferably, the radionuclide and peptide are linked by a chelating agent (e.g., DOTA or NOA).

[0125] In one embodiment, the radionuclide is selected from the group consisting of therapeutic alpha emitters, therapeutic beta emitters, or therapeutic Auger emitters.

[0126] In another embodiment, the radionuclide is used for targeted radionuclide therapy (alpha, beta emitters, or Auger), and is selected from the group consisting of the following isotopes: 67 Cu、 177 Lu、 89 Sr、 90 Y、 117 mSn, 131 I, 153 Sm、 166 Ho、 186 Re、 188 Re、 211 At、 212 Pb, 212 Bi、 213 Bi、 22 3Ra、 224 Ra、 225 Ac、 227 Th, preferably selected from 177 Lu、 67 Cu、 90 Y、 211 At、 225 Ac and 227 Th, more preferably 177 Lu.

[0127] In another embodiment, the radiopharmaceutical is conjugated to the uPAR-binding peptide via a chelating agent, such as a chelating agent selected from the group consisting of: DOTA, CB-DO2A, 3p-C-DEPA, TCMC, Oxo-DO3A, TETA, TE2A, CB-TE2A, CB-TE1A1P, CB-TE2P, MM-TE2A, DM-TE2A, SarAr, SarAr-NCS, diamSar, AmBaSar, BaBaSar, ATSM, CB-TE1A1P and CB-TE2P, Nota, NETA, TACN-TM, NODAGA, TRAP, AAZTA, DATA, H2dedpa, CP256, PCTA, THP, DTPA, 1B4M-DTPA, CHX-A′′-DTPA, TRAP (PRP9), NOPO, DFO HOPO, H6phospa, PCTA, H2dedpa, H4octapa, H2azapa, H5decapa, HBED, HBED-cc, SHBED, BPCA, CP256, HEHA, PEPA and RESCA1, preferably selected from any one of DOTA, NOTA, CB-TE2A, NODAGA, DFO, HBED and HBED-cc, more preferably, the chelating agent is DOTA or NOTA.

[0128] In one implementation scheme, the radiopharmaceutical is selected from... 177 Lu-DOTA-AE105 67 Cu-DOTA-AE105 225 Ac-DOTA-AE105 and 212 The group consists of Pb-DOTA-AE105.

[0129] In one implementation, the radiopharmaceutical (radioactive nuclide and chelating agent) is selected from... 177 Lu-DOTA, 67 Cu-DOTA, 225 Ac-DOTA and 212 A group consisting of Pb-DOTA.

[0130] In a preferred embodiment, the radiopharmaceutical is 177 Lu-DOTA-AE105.

[0131] In another preferred embodiment, the radiopharmaceutical is 67 Cu-DOTA-AE105.

[0132] In yet another embodiment, the radiopharmaceutical has the following formula: As outlined in the data discussion below, the findings presented in this paper highlight the potential of uPAR as a therapeutic target for glioma, and more importantly, as a target for uPAR-PRRT. In particular, positive uptake of uPAR-PET suggests that uPAR-PRRT labeled with therapeutic α or β emitters can be administered systemically, rather than intratumorally.

[0133] Therefore, in one implementation, the radiopharmaceutical is administered systemically or intratumorally.

[0134] In another implementation, the radiopharmaceutical is administered systemically.

[0135] In one embodiment, the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma (WHO grade 4) (glioblastoma).

[0136] In another implementation, the brain tumor is a high-grade glioma (glioblastoma).

[0137] Note again that embodiments of other aspects of the invention are also applicable to this aspect.

[0138] All patent and non-patent references cited in this application are incorporated herein by reference in their entirety.

[0139] The invention will now be described in more detail in the following non-limiting embodiments.

[0140] Example Example 1 – Materials and Methods Research Design In this prospective clinical trial, eligible patients were enrolled from the Department of Neurosurgery at Rigshospitalet, University Hospital of Copenhagen, between March 2017 and June 2022. Eligibility was determined by meeting the following inclusion criteria: being over 18 years of age, being able to read and understand patient information in Danish and providing informed consent; having a newly diagnosed intracranial lesion suspected to be a primary glioma on brain MRI; and planning to undergo neurosurgery (biopsy or tumor resection).

[0141] If the patient is pregnant or breastfeeding, weighs more than 140 kg, has claustrophobia, is over 85 years old, or is suspected of having claustrophobia... 68 Allergy to Ga-NOTA-AE105 is excluded.

[0142] Written informed consent was obtained from all patients. Following consent, patients were scheduled to undergo neurosurgical procedures. 68 Ga-NOTA-AE105 PET / MRI brain scan.

[0143] PET / MRI acquisition As mentioned above, synthesize tracers. 68 Ga-NOTA-AE105(13). PET / MRI scans were performed using an integrated PET / MRI system (Siemens Biograph mMR; Siemens Healthcare) with a radiotracer. The injection volume was approximately 200 MBq (median: 202; range: 83–222 MBq). 68 Following the Ga-NOTA-AE105, a dynamic 60-minute PET / MRI scan was performed.

[0144] If a patient was ineligible for PET / MRI due to contraindications, a PET / CT scan was performed using a Biograph 128 mCT PET / CT device (Siemens Medical Solutions) with an axial field of view of 21.6 cm. However, of the 24 patients available for the final analysis, only one underwent PET / CT instead of PET / MRI (see below).

[0145] PET images were reconstructed using a deep learning-based pseudo-CT attenuation map (22) based on a UTE MRI sequence with absolute scattering correction (3D ordinary Poisson-ordered subset expectation maximization [3D-OP-OSEM], 4 iterations, 21 subsets, 3.5 mm Gaussian filter). The images were obtained from an injection... 68 Static images were reconstructed from data acquired in a series of data at 20-40 min, 40-60 min, and dynamic 0-60 min after Ga-NOTA-AE105 injection. Reconstructed PET-MRI images at 20-40 min after tracer injection were used for further interpretation, quantification, and analysis.

[0146] MRI protocol The MRI scanning protocol included UTE AC sequence, 3D T1-weighted (T1W) MPRAGE with gadolinium injection before and after contrast administration, T2-weighted (T2W) dark fluid turbine inversion recovery amplitude (TIRM) (FLAIR) in the axial and coronal planes, diffusion-weighted (DWI) RESOLVE, and T2W BLADE. Parameters are listed in Table 1.

[0147] Table 1. MR parameters Image analysis Analysis of the reconstructed image data was performed independently by certified nuclear medicine specialists and certified neuroradiologists. Each specialist was unaware of the clinical data. Tumors were delineated by plotting regions of interest (VOIs) on PET images, and maximum normalized uptake values ​​(SUV maxima) were measured. If uPAR-positive lesions were not visible on PET images, tumors were delineated using MRI or CT images for SUV maximization measurements. Reference brain VOIs parallel to the cortex were plotted on single-layer slices at the level of the centrum semiovale on the contralateral normal cerebral hemisphere. The VOIs were positioned approximately 7 mm from the cortical margin to avoid spillage of blood pool activity, and mean normalized uptake values ​​(SUV mean) were measured. Lesions were considered uPAR-positive if the ratio of the tumor SUV maxima to the mean normal brain SUV (TBR) was at least 2.0, as used in the previous uPAR-PET study (17). Tumor size was measured on axial T2W FLAIR MRI or axial CT images according to the Neuro-oncology Response Assessment (RANO) criteria (23), as the product of the maximum vertical diameter. If the lesion is not visible on the CT image, the tumor size is measured using the most recent MRI scan that is closest to the PET / CT scan.

[0148] Follow-up These patients received routine follow-up at the Rigshospitalet Oncology Department of Copenhagen University Hospital. The follow-up protocol was standardized according to the Danish National Guidelines for Glioma published by the Danish Neuro-Oncology Group (DNOG) (24). The final follow-up endpoint was performed on November 29, 2022. Progression-free survival (PFS) was assessed using the RANO criteria and defined as the time from uPAR-PET / MRI scan to progression (23, 25). Overall survival (OS) was defined as the time from uPAR-PET / MRI scan to death. Patients who had not progressed at follow-up were censored based on the date of their most recent clinical follow-up.

[0149] Statistical analysis Sample size was calculated based on disease prevalence and a 36-month follow-up period. A total of 29 patients were required to detect significant differences in PFS and OS (Type I error risk of 0.05, power of 0.8). Considering potential dropouts, a target of 30–35 patients was planned for the trial. All continuous variables were reported as means with standard deviation (SD) or medians with ranges. Kaplan-Meil analysis was performed to estimate PFS and OS, and reverse Kaplan-Meil analysis was performed to estimate median follow-up time. Univariate Cox regression analysis was performed on OS and PFS with uPAR SUV maximum as a continuous variable. To determine the optimal cutoff value for uPAR SUV maximum 20–40 min post-traumatic injection, the inventors used the Cut-off Finder application (26). A p-value less than 0.05 was considered statistically significant. Data analysis was performed using R version 4.2.2 (R Foundation for Statistical Computing).

[0150] Example 2 - Patient and Image Acquisition From March 2017 to June 2022, a total of 33 patients were enrolled in the trial. Of these, 29 patients underwent dynamic PET / MRI (n=26) or PET / CT (n=3) brain scans. Four patients were excluded due to radiopharmaceutical production failure (n=3) and technical issues (n=1). Data from 27 of these patients were available for reconstruction. Histological examination was performed on all 27 patients, and they were reviewed according to the 2021 WHO classification of diseases of the central nervous system (2). Three patients were excluded because they were diagnosed with central nervous system lymphoma. Therefore, the final trial population consisted of 24 patients diagnosed with primary glioma, of whom 23 underwent PET / MRI and 1 underwent PET / CT, see [link to relevant section]. Figure 1 .

[0151] Table 2 summarizes the demographic data of the 24 patients.

[0152] Table 2. Baseline Characteristics Most patients were diagnosed with WHO grade 4 glioma (67%, 16 / 24), followed by grade 3 (25%, 6 / 24) and grade 2 (8%, 2 / 24). The majority of tumors were located in the corpus callosum (21%, 5 / 24), frontal lobe (25%, 6 / 24), or temporal lobe (21%, 5 / 24). No patient presented worse than WHO grade 1. The median tumor size was 1,700 mm² (range: 320–3,220 mm²). The median time from PET / MRI scan to surgery was 1 day (range: 0–21 days). [Tracer] 68 The median injection dose of Ga-NOTA-AE105 was 5.0 ml (range, 0.3 ml–7.5 ml), and the median activity was 202 MBq (range, 83–222 MBq). No adverse events or serious adverse events were recorded during the study.

[0153] in conclusion uPAR-PET imaging is feasible and safe for patients with gliomas.

[0154] Example 3 - Image Analysis Of the 24 patients, 16 (67%, 16 / 24) were PET-positive. Among the PET-positive patients, 15 (94%, 15 / 16) showed contrast enhancement on MRI, while 1 (6%, 1 / 16) did not (4%, 1 / 24). Of the 24 patients, 8 (33%, 8 / 24) were PET-negative. Among the PET-negative patients, 8 (100%, 8 / 8) showed no pathological contrast enhancement. uPAR-positive lesions were mainly seen in WHO grade 4 gliomas (94%, 15 / 16), including one patient with a WHO grade 3 glioma who also had a PET-positive tumor. Figure 2 and Figure 3 This shows a representative example of a PET-positive tumor lesion.

[0155] in conclusion Therefore, uPAR PET can identify high-grade gliomas (WHO grades 3 and 4) and demonstrates the high uptake and applicability of uPAR-targeted radionuclide therapy.

[0156] Example 4 - Follow-up The median follow-up time from uPAR-PET / MRI scan to PFS, OS, or patient cleavage was 9.8 (IQR, 7.2–26.4) months. A total of 16 patients (67%) experienced disease progression (15 grade 4 and 1 grade 2), and 10 patients (42%) died (all grade 4). Table 2 above shows the first-line surgical and oncology treatments during follow-up. All patients underwent surgery, with over half undergoing surgical resection (54%, 13 / 24), and the remaining patients undergoing biopsy (46%, 11 / 24). The most common oncology treatment was a combination of radiotherapy and chemotherapy (54%, 13 / 24); however, some patients received only radiotherapy (38%, 9 / 24) or no adjuvant therapy (8%, 2 / 24).

[0157] in conclusion Patients are followed up for a longer period of time, and usually undergo preliminary treatment with a combination of surgery, radiotherapy and chemotherapy.

[0158] Example 5 - Progression-free survival and overall survival Research Objective Progression-free survival (PFS) and overall survival (OS) were determined based on uPAR imaging.

[0159] result Using the Cut-off Finder program, the optimal cutoff points for OS and PFS for the maximum SUV value in all primary glioma groups (n=24) were 1.1 for OS and 0.64 for PFS.

[0160] Using these cutoff values, uPAR expression was categorized into high and low expression. High expression indicated significantly worse prognoses in terms of overall survival (OS) and progression-free survival (PFS), with HRs of 10.5 (95% CI, 1.31–83.1; P = 0.027) and 17.3 (95% CI, 2.22–134.0; P = 0.0064), respectively. Figure 4A and 4B ).

[0161] uPAR expression, as a continuous variable, was also associated with poorer prognoses in OS and PFS, with HRs of 2.48 (95% CI, 1.26–4.88; P = 0.0084) and 2.28 (95% CI, 1.35–3.86; P = 0.0020), respectively.

[0162] Additional subgroup analyses based solely on major HGG (n=22) were also performed. For the high-grade group, the optimal cutoff points for OS and PFS based on maximum SUV were both 1.1. Within this subgroup, uPAR expression was also categorized as high or low; high uPAR uptake was significantly worse in terms of PFS compared to low uPAR uptake (HR 13.4, 95% CI, 1.70–102.0; P=0.014), while OS was critically significantly worse (HR 7.44, 95% CI, 0.94–59.0; P=0.058). Figure 5A and 5B ).

[0163] Furthermore, analysis of uPAR expression as a continuous variable in the HGG subgroup was also associated with poorer prognoses in OS and PFS, with HRs of 2.23 (95% CI, 1.11–4.50; P = 0.025) and 2.11 (95% CI, 1.24–3.61; P = 0.0063), respectively.

[0164] in conclusion uPAR-PET has been found to be a powerful prognostic assessment tool.

[0165] Example 6 - Image Analysis and Comparison with Somatostatin Receptor Imaging Surprisingly, even when MRI was available, there was no perfect correlation between MRI contrast enhancement and high uptake by uPAR-PET, demonstrating the value of uPAR-PET. Furthermore, uPAR-PET was meaningful as a continuous variable for OS and PFS, demonstrating the difference compared to MRI contrast enhancement (divided into enhanced and non-enhanced).

[0166] Data discussion The current study found that tumor SUVs 最大值 The measured uPAR-PET activity predicted poor overall survival (OS) and progression-free survival (PFS) outcomes in patients with primary gliomas. This effect can likely be attributed to the difference in expected survival between uPAR-negative LGG and HGG, which constitute the majority of the inventors' cohort. However, even analyzing uPAR-PET for HGG alone still has prognostic significance. Therefore, uPAR-PET can be used for prognostic assessment and treatment planning, such as surgical strategies for these patients. Furthermore, the inventors found that uPAR-PET was positive in the majority (67%) of glioma patients, which may encourage further development of uPAR-PRRT for glioma patients.

[0167] In summary, these findings highlight the potential of uPAR as a therapeutic target for glioma, and most importantly, its potential as a target for uPAR-PRRT. In particular, it should be noted that positive uptake by uPAR-PET suggests that uPAR-PRRT, using similar ligands but labeled with therapeutic α or β emitters, can be administered systemically rather than intratumorally.

[0168] It should be noted that external beam radiation therapy (EPR) is well-established in the treatment of HGG, paving the way for targeted radioligand therapy in these patients. PRRT for brain tumors, as a highly localized treatment, is superior to the less precise EPR because it may reduce the well-known cognitive side effects associated with EPR due to whole-brain irradiation. Targeted radioligand therapy against the somatostatin receptor (SSTR-PRRT) is primarily used for neuroendocrine tumors and has also been applied in gliomas. SSTR expression has been reported in approximately 25% of gliomas, with differences in expression between LGG and HGG, but decreased SSTR2 expression is observed in the most aggressive gliomas (27). In contrast, the inventors found that 94% of WHO grade 4 tumors were positive for uPAR-PET.

[0169] One study investigated PRRT for SSTRs, in which 10 patients with WHO grade 2-3 gliomas received intratumoral injections. 90 Y-DOTATOC treatment. It has been reported that... 90 Y-DOTATOC treatment safely and effectively halted tumor progression for at least 13–45 months (28). Subsequently, another study demonstrated similar efficacy in 2010. 90 The safety and efficacy of Y-DOTATOC in treating three patients with recurrent glioblastoma (29). In recent years, there has been increasing interest in α-emitter PRRT targeting the neurokinin type 1 receptor (NK1R) (30). Interestingly, α-emitter PRRT was used in nine patients with recurrent glioblastoma. 213 PRRT therapy with intratumoral administration of Bi-DOTA substance P has been shown to be safe (31). Therefore, PRRT for glioma has been thoroughly studied, and to date, intratumoral α-emitter PRRT has been reported to be safe, feasible, and effective in promoting clinically meaningful responses in multiple clinical studies, highlighting the prospect of PRRT as an alternative to traditional therapies for glioma.

[0170] uPAR shows promise for targeted cancer therapy due to its central role in tumor invasion and metastasis. One reason behind this is the conceptual advantage of targeting a receptor that is primarily overexpressed in the most aggressive and invasive regions of a tumor. In this study, the inventors found that most patients exhibited uPAR expression, and that uPAR expression was associated with worsening outcomes, data supported by existing literature that found high uPAR expression, particularly in HGG, and associated with poor prognosis (32). This highlights the role of uPAR as an ideal target, expressed in most HGG, where treatment can target the most aggressive parts of the tumor. Several uPAR-targeting therapies have been or are under investigation but have not yet been approved for clinical use (33, 34). The inventors' team published a paper on the use of... 177 A preclinical paper on Lu-DOTA-AE105-targeted uPAR-based PRRT for colorectal cancer xenograft (20).

[0171] In this study, the inventors demonstrated a significant reduction in tumor size and good tolerability in mice. Similarly, the inventors demonstrated... 177 The efficacy of Lu-DOTA-AE105 in treating a model of disseminated metastatic prostate cancer (21). Therefore, PRRT targeting uPAR appears to have great potential in several tumor types, but further investigation in a clinical setting is needed. 177 One advantage of using Lu-DOTA-AE105 for PRRT is that it is based on... 68 Ga-NOTA-AE105 is the same uPAR-binding peptide AE105, which means that uPAR-PET can be used as a companion diagnostic agent for treatment planning, monitoring and dose estimation in the diagnostic approach of uPAR-PRRT therapy for glioma.

[0172] While prolonged OS and PFS are the desired goals of PRRT for glioma patients, alternatives to external beam radiation therapy can reduce side effects on the healthy brain due to their more specific tumor tissue targeting.

[0173] in conclusion The inventors demonstrated that uPAR expression, as measured by uPAR-PET, was significantly associated with worse overall survival (OS) and progression-free survival (PFS) in patients with primary glioma, indicating that uPAR radiotracers (using uPAR-PET) are effective in preventing the spread of the virus. 68 The prognostic value of uPAR (taking Ga-NOTA-AE105 as an example) is highlighted. This underscores uPAR's promising target for diagnosis, prognostic assessment, and targeted therapy for gliomas. Most importantly, uPAR holds significant potential as a therapeutic target for PRRT, with uPAR-PET serving as a companion diagnostic in therapeutic diagnostics to preselect uPAR-PRRT patients.

[0174] References 1. Ostrom QT, Cioffi G, Gittleman H, et al. CBTRUS Statistical Report: Primary Brain and Other Central Nervous System Tumors Diagnosed in the United States in 2012 - 2016. Neuro Oncol. 2019;21:v1 - v100. 2. Louis DN, Perry A, Wesseling P, et al. The 2021 WHO Classification of Tumors of the Central Nervous System: a summary. Neuro Oncol. 2021;23:1231 - 1251. 3. Weller M, van den Bent M, Preusser M, et al. EANO guidelines on the diagnosis and treatment of diffuse gliomas of adulthood. Nat Rev Clin Oncol. 2021;18:170 - 186. 4. Rasmussen BK, Hansen S, Laursen RJ, et al. Epidemiology of glioma: clinical characteristics, symptoms, and predictors of glioma patients grade I - IV in the the Danish Neuro - Oncology Registry. J Neurooncol. 2017;135:571 - 579. 5. Stupp R, Mason WP, van den Bent MJ, et al. Radiotherapy plus concomitant and adjuvant temozolomide for glioblastoma. N Engl J Med. 2005;352:987 - 996. 6.Stupp R, Hegi ME, Mason WP, et al. Effects of radiotherapy withconcomitant and adjuvant temozolomide versus radiotherapy alone on survivalin glioblastoma in a randomised phase III study: 5-year analysis of theEORTC-NCIC trial. Lancet Oncol. 2009;10:459-466. 7.Ellingson BM, Wen PY, Cloughesy TF. Modified Criteria forRadiographic Response Assessment in Glioblastoma Clinical Trials.Neurotherapeutics. 2017;14:307-320. 8.Albert NL, Weller M, Suchorska B, et al. Response Assessment inNeuro-Oncology working group and European Association for Neuro-Oncologyrecommendations for the clinical use of PET imaging in gliomas. Neuro Oncol.2016;18:1199-1208. 9.Law I, Albert NL, Arbizu J, et al. Joint EANM / EANO / RANO practiceguidelines / SNMMI procedure standards for imaging of gliomas using PET withradiolabelled amino acids and [(18)F]FDG: version 1.0. Eur J Nucl Med MolImaging. 2019;46:540-557. 10.Mahmood N, Rabbani SA. Fibrinolytic System and Cancer: Diagnosticand Therapeutic Applications. Int J Mol Sci. 2021;22. 11.Ahn SB, Mohamedali A, Pascovici D, et al. Proteomics Reveals Cell-Surface Urokinase Plasminogen Activator Receptor Expression Impacts MostHallmarks of Cancer. Proteomics. 2019;19:e1900026. 12.Mahmood N, Mihalcioiu C, Rabbani SA. Multifaceted Role of theUrokinase-Type Plasminogen Activator (uPA) and Its Receptor (uPAR):Diagnostic, Prognostic, and Therapeutic Applications. Front Oncol. 2018;8:24. 13.Skovgaard D, Persson M, Brandt-Larsen M, et al. Safety, Dosimetry,and Tumor Detection Ability of (68)Ga-NOTA-AE105: First-in-Human Study of aNovel Radioligand for uPAR PET Imaging. J Nucl Med. 2017;58:379-386. 14.Persson M, Madsen J, Østergaard S, Ploug M, Kjaer A. 68Ga-labelingand in vivo evaluation of a uPAR binding DOTA- and NODAGA-conjugated peptidefor PET imaging of invasive cancers. Nucl Med Biol. 2012;39:560-569. 15.Kriegbaum MC, Persson M, Haldager L, et al. Rational targeting ofthe urokinase receptor (uPAR): development of antagonists and non-invasiveimaging probes. Curr Drug Targets. 2011;12:1711-1728. 16.Fosbøl M, Kurbegovic S, Johannesen HH, et al. Urokinase-TypePlasminogen Activator Receptor (uPAR) PET / MRI of Prostate Cancer forNoninvasive Evaluation of Aggressiveness: Comparison with Gleason Score in aProspective Phase 2 Clinical Trial. J Nucl Med. 2021;62:354-359. 17.Carlsen EA, Loft M, Loft A, et al. Prospective Phase II Trial ofPrognostication by (68)Ga-NOTA-AE105 uPAR PET in Patients with NeuroendocrineNeoplasms: Implications for uPAR-Targeted Therapy. J Nucl Med. 2022;63:1371-1377. 18.Risør LM, Clausen MM, Ujmajuridze Z, et al. Prognostic Value ofUrokinase-Type Plasminogen Activator Receptor PET / CT in Head and NeckSquamous Cell Carcinomas and Comparison with (18)F-FDG PET / CT: A Single-Center Prospective Study. J Nucl Med. 2022;63:1169-1176. 19.Persson M, Nedergaard MK, Brandt-Larsen M, et al. Urokinase-TypePlasminogen Activator Receptor as a Potential PET Biomarker in Glioblastoma.J Nucl Med. 2016;57:272-278. 20.Persson M, Rasmussen P, Madsen J, Ploug M, Kjaer A. New peptidereceptor radionuclide therapy of invasive cancer cells: in vivo studies using177Lu-DOTA-AE105 targeting uPAR in human colorectal cancer xenografts. NuclMed Biol. 2012;39:962-969. 21.Persson M, Juhl K, Rasmussen P, et al. uPAR targeted radionuclidetherapy with (177)Lu-DOTA-AE105 inhibits dissemination of metastatic prostatecancer. Mol Pharm. 2014;11:2796-2806. 22.Ladefoged CN, Hansen AE, Henriksen OM, et al. AI-drivenattenuation correction for brain PET / MRI: Clinical evaluation of a dementiacohort and importance of the training group size. Neuroimage. 2020;222:117221. 23.Leao DJ, Craig PG, Godoy LF, Leite CC, Policeni B. ResponseAssessment in Neuro-Oncology Criteria for Gliomas: Practical Approach UsingConventional and Advanced Techniques. AJNR Am J Neuroradiol. 2020;41:10-20. 24.DNOG. The Danish Neuro-Oncology Group. Gliomas in adults [18 / 12 / 2020; https: / / dnog.dk / assets / files / Retningslinier%20PDF / DNOG_gliomer_voksne_AdmGodk181220.pdf. Accessed 04 / 04 / 2023. 25.Wen PY, Macdonald DR, Reardon DA, et al. Updated responseassessment criteria for high-grade gliomas: response assessment in neuro-oncology working group. J Clin Oncol. 2010;28:1963-1972. 26.Budczies J, Klauschen F, Sinn BV, et al. Cutoff Finder: acomprehensive and straightforward Web application enabling rapid biomarkercutoff optimization. PLoS One. 2012;7:e51862. 27.Lange F, Kaemmerer D, Behnke-Mursch J, Brück W, Schulz S, Lupp A.Differential somatostatin, CXCR4 chemokine and endothelin A receptorexpression in WHO grade I-IV astrocytic brain tumors. J Cancer Res ClinOncol. 2018;144:1227-1237. 28.Schumacher T, Hofer S, Eichhorn K, et al. Local injection of the90Y-labelled peptidic vector DOTATOC to control gliomas of WHO grades II andIII: an extended pilot study. Eur J Nucl Med Mol Imaging. 2002;29:486-493. 29.Heute D, Kostron H, von Guggenberg E, et al. Response of recurrenthigh-grade glioma to treatment with (90)Y-DOTATOC. J Nucl Med. 2010;51:397-400. 30.Cimini A, Ricci M, Russo F, et al. Peptide Receptor RadionuclideTherapy and Primary Brain Tumors: An Overview. Pharmaceuticals (Basel). 2021;14. 31.Krolicki L, Bruchertseifer F, Kunikowska J, et al. Prolongedsurvival in secondary glioblastoma following local injection of targetedalpha therapy with (213)Bi-substance P analogue. Eur J Nucl Med Mol Imaging.2018;45:1636-1644. 32.Hirata K, Tamaki N. uPAR as a Glioma Imaging Target. Journal ofNuclear Medicine. 2016;57:169-170. 33.Mahmood N, Arakelian A, Khan HA, Tanvir I, Mazar AP, Rabbani SA.uPAR antibody (huATN-658) and Zometa reduce breast cancer growth and skeletallesions. Bone Res. 2020;8:18. 34.Heinemann V, Ebert MP, Laubender RP, Bevan P, Mala C, Boeck S.Phase II randomised proof-of-concept study of the urokinase inhibitorupamostat (WX-671) in combination with gemcitabine compared with gemcitabinealone in patients with non-resectable, locally advanced pancreatic cancer. BrJ Cancer. 2013;108:766-770. 35. Kurbegovic S, Juhl K, Sørensen KK, et al. IRDye800CW labeled uPAR-targeting peptide for fluorescence-guided glioblastoma surgery: Preclinical studies in orthotopic xenografts. Theranostics. 2021;11:7159-7174. 36. Persson M, Skovgaard D, Brandt-Larsen M, Christensen C, Madsen J, Nielsen CH, Thurison T, Klausen TL, Holm S, Loft A, Berthelsen AK, Ploug M, Pappot H, Brasso K, Kroman N, Højgaard L, Kjaer A. First-in-human uPAR PET: Imaging of Cancer Aggressiveness. Theranostics. 2015 Sep 13;5(12):1303-16.

Claims

1. A positron emission tomography (PET) imaging agent for prognostic assessment of progression-free survival (PFS) and / or overall survival (OS) in patients with brain tumors via PET imaging of cancer. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 maximum and / or average SUV levels above the threshold indicate poor prognosis in PFS and / or OS; and The maximum SUV level and / or the average SUV level, which are equal to or below the threshold level, indicate a good prognosis for PFS and / or OS.

2. The imaging agent used according to claim 1, wherein the peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser).

3. The developing agent according to claim 1 or 2, wherein the radionuclide is selected from... 68 Ga and 64 The group consisting of Cu.

4. The imaging agent used according to any one of the preceding claims, wherein the radionuclide is coupled to the uPAR-binding peptide by a chelating agent such as DOTA or NOA.

5. The developing agent used according to any one of the preceding claims, wherein the radionuclide is 68 Ga, which is coupled to the uPAR-binding peptide via the chelating agent NOA.

6. The developer used according to any one of the preceding claims, having the following formula: 。 7. The developing agent used according to any one of claims 1-4, wherein the radionuclide is 64 Cu, which is coupled to the uPAR-binding peptide via a chelating agent DOTA or NOA.

8. The imaging agent used according to any one of the preceding claims, wherein the imaging agent is administered at a dose of 10-500 MBq, and then a PET scan is performed 10 minutes to 24 hours after the administration of the imaging agent, and quantified by the maximum SUV value and / or the average SUV value.

9. The developer used according to any one of the preceding claims, wherein - For all gliomas, the maximum and / or average threshold values ​​of the SUV associated with PFS are in the range of 0.3-1, preferably 0.4-0.8, more preferably about 0.64; and / or - For all gliomas, the threshold values ​​of maximum and / or average SUV associated with OS are in the range of 0.7-1.5, preferably 0.9-1.3, and more preferably about 1.

1.

10. The developer used according to any one of claims 1-8, wherein - For high-grade gliomas, the maximum and / or average threshold values ​​of the SUV associated with PFS are in the range of 0.7-1.5, preferably 0.9-1.3, more preferably about 1.1; and / or - For all high-grade gliomas, the threshold values ​​of maximum SUV and / or average SUV associated with OS are in the range of 0.7-1.5, preferably 0.9-1.3, and more preferably about 1.

1.

11. The imaging agent used according to any one of the preceding claims, wherein the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma (WHO grade 4) (glioblastoma).

12. The imaging agent used according to any one of the preceding claims, wherein the brain tumor is a high-grade glioma (glioblastoma).

13. A positron emission tomography (PET) imaging agent used as a companion diagnostic agent for patients with brain tumors via PET imaging for cancer. The imaging agent contains a uPAR-binding peptide conjugated to a radionuclide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or its uPAR-binding variant; in The maximum and / or average SUV levels above the threshold indicate that uPAR-binding drugs, such as uPAR-binding radiopharmaceuticals, will be effective against the brain tumor; and The maximum and / or average SUV levels, which are equal to or below the threshold level, indicate that uPAR drugs, such as uPAR combined with radiopharmaceuticals, will be ineffective against the brain tumor. The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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).

14. The positron emission tomography (PET) imaging agent according to claim 13, wherein the radionuclide and the chelating agent are... 68 Ga-NOTA.

15. The positron emission tomography (PET) imaging agent according to claim 13, wherein the radionuclide and the chelating agent are... 64 Cu-DOTA.

16. The positron emission tomography (PET) imaging agent used according to any one of claims 13-15, wherein it is a companion diagnostic reagent for a radiopharmaceutical.

17. The positron emission tomography (PET) imaging agent used according to any one of claims 13-16, wherein it is a companion diagnostic reagent for a radiopharmaceutical as defined in any one of claims 24-30.

18. The positron emission tomography (PET) imaging agent used according to claim 17, which is a companion diagnostic reagent for a radiopharmaceutical as defined in claim 24, wherein the radionuclide and chelating agent of the radiopharmaceutical are... 177 Lu-DOTA.

19. The positron emission tomography (PET) imaging agent used according to any one of claims 13-18, wherein it is a companion diagnostic reagent for a radiopharmaceutical having the following formula: 。 20. The positron emission tomography (PET) imaging agent used according to any one of claims 13-19, wherein the companion diagnostic agent is used to determine the suitability of a radiopharmaceutical, such as a radiopharmaceutical as defined in any one of claims 24-30, for a particular individual.

21. The positron emission tomography (PET) imaging agent used according to any one of claims 13-20, wherein the companion diagnostic reagent is used to determine the suitability of the radiopharmaceutical as defined in any one of claims 24-30.

22. The positron emission tomography (PET) imaging agent used according to any one of claims 13-21, wherein the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma (WHO grade 4) (glioblastoma).

23. The positron emission tomography (PET) imaging agent used according to any one of claims 13-22, wherein the brain tumor is a high-grade glioma (glioblastoma).

24. A composition comprising a radiopharmaceutical for treating or alleviating a brain tumor in a subject; The radiopharmaceutical contains a radionuclide and a uPAR-binding peptide; and The uPAR-binding peptide is (D-Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(Tyr)-(Leu)-(Trp)-(Ser) or a uPAR-binding variant thereof; and The uPAR binding variants are selected from the group consisting of the following: (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]ethoxyethyl)glycine), (Asp)-([β]-cyclohexyl-L-alanine)-(Phe)-(D-Ser)-(D-Arg)-(N-(2,3-dimethoxybenzyl)glycine)-(D-Phe)-(N-(methylnaphthyl)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 subjects in this study have been diagnosed with brain tumors that express uPAR, and uPAR expression is above a predetermined threshold level. The threshold level is determined using a positron emission tomography (PET) agent as defined in any one of claims 1-12.

25. The composition according to claim 24, wherein the radionuclide is used for targeted radionuclide therapy and is selected from the group consisting of: 67 Cu、 177 Lu、 89 Sr、 90 Y、 117 mSn, 131 I, 153 Sm、 166 Ho、 186 Re、 188 Re、 211 At、 212 Pb, 212 Bi、 213 Bi、 223 Ra、 224 Ra、 225 Ac、 227 Th, preferably selected from 177 Lu、 67 Cu、 90 Y、 211 At、 225 Ac and 227 Th, more preferably 177 Lu.

26. The composition used according to claim 24 or 25, wherein the radiopharmaceutical is conjugated to the uPAR-binding peptide by a chelating agent, for example, the chelating agent being selected from the group consisting of: DOTA, CB-DO2A, 3p-C-DEPA, TCMC, Oxo-DO3A, TETA, TE2A, CB-TE2A, CB-TE1A1P, CB-TE2P, MM-TE2A, DM-TE2A, SarAr, SarAr-NCS, diamSar, AmBaSar, BaBaSar, ATSM, CB-TE1A1P and CB-TE2P, Nota, NETA, TACN-TM, NODAGA, TRAP, AAZTA, DATA, H2dedpa, CP256, PCTA, THP, DTPA, 1B4M-DTPA, CHX-A′′-DTPA, TRAP (PRP9), NOPO, DFO HOPO, H6phospa, PCTA, H2dedpa, H4octapa, H2azapa, H5decapa, HBED, HBED-cc, SHBED, BPCA, CP256, HEHA, PEPA and RESCA1, preferably selected from any one of DOTA, NOTA, CB-TE2A, NODAGA, DFO, HBED and HBED-cc, more preferably, the chelating agent is DOTA or NOTA.

27. The composition used according to any one of claims 24-26, wherein the radionuclide and the chelating agent are 177 Lu-DOTA.

28. The composition according to any one of claims 24-27, wherein the radiopharmaceutical has the following formula: 。 29. The positron emission tomography (PET) imaging agent used according to any one of claims 24-28, wherein the brain tumor is a high-grade glioma (WHO grade 3 and 4) or a low-grade glioma (WHO grade 1 and 2), preferably a high-grade glioma (WHO grade 4) (glioblastoma).

30. The positron emission tomography (PET) imaging agent used in any one of claims 24-29, wherein the brain tumor is a high-grade glioma (glioblastoma).

Images