Therapeutic application of 64cu for radionuclide therapy

Abstract

The present invention relates to application of 64Cu in radionuclide-based cancer therapy in humans. More specifically, the invention relates to 64Cu labeled conjugates for use in the treatment of cancer patients including 64Cu labelled Integrin αVβ3 binding conjugates, such as [64Cu]NODAGA-E[c(RGDyK)]2.

Description

[0001] Therapeutic application of64Cu for Radionuclide Therapy

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to application of 64Cu in radionuclide-based cancer therapy in humans. More specifically, the invention relates to 64Cu labeled conjugates for use in the treatment of cancer patients including 64Cu labelled Integrin aV 3 binding conjugates, such as 64CuNODAGA-E[c(RGDyK)]2.

[0004] BACKGROUND OF THE INVENTION

[0005] Various radionuclide labelled targeting moieties, such as peptide compositions, have been developed or are under development for site-specific targeting of a therapeutic radionuclide. The general principle involves attaching a selected radionuclide to a targeting moiety, such as a peptide, having a high specificity for a particular organ or tissue so that the organ or tissue can be treated by a therapeutic radioisotope. This field of research has shown promising applicability for tumor treatment.

[0006] Therapeutic radionuclides are radioactive isotopes having a set of properties that will deliver targeted radiation to diseased tissues, with minimal damage to surrounding healthy tissues. In order to achieve localized radiation, a key feature of a therapeutic radionuclide is the type of radiation emitted. Therapeutic radionuclides emit high-linear energy transfer (LET) radiation typically in the form of beta minus (P-) particles or alpha minus (a-) particles providing a high ionization per length of travel, which is fully deposited within a small range of tissue, usually in the range of mm, thereby minimizing damage of surrounding healthy tissue. Beta minus emitters, such as Yttrium-90 (90Y), Copper-67 (67Cu), and Lutetium-177 (177Lu), are commonly used due to their moderate tissue penetration and suitable half-lives. Alpha emitters, like Actinium-225 (225Ac) and Radium-223 (223Ra), provide high linear energy transfer (LET), which results in significant cytotoxicity within a short range, making them ideal for targeting small clusters of cancer cells.

[0007] Furthermore, relevant features for the selection of an appropriate radionuclide for therapy also include the radionuclide half-life and ability to be conjugated to targeting molecules, such as peptides. The half-life of a radionuclide influences its clinical applicability. It must be long enough to allow for manufacturing, quality control, and administration, yet short enough to minimize unnecessary radiation exposure to the patient. For example, Yttrium-90 (90Y) has a half-life of 64.1 hours, Copper-67 (67Cu) has a half-life of 2.6 days, Lutetium-177 (177Lu) has a half-life of 6.7 days, Radium-223 (223Ra) has a half-life of 11.4 days, and Actinium-225 (225Ac) has a half-life of 10 days.

[0008] Radionuclides used in medical imaging possess distinct physical and chemical properties that make them suitable for diagnostic purposes. Their efficacy in imaging relies on several key characteristics, including half-life, type of emitted radiation, energy of radiation, and chemical compatibility with biological systems. The half-life of a radionuclide is critical in imaging as it determines the duration of its activity within the body. Ideal radionuclides for diagnostic imaging have a half-life that is long enough to perform the imaging procedure but short enough to minimize radiation exposure to the patient. For imaging purposes, gamma-emitting radionuclides are primarily used due to their ability to penetrate tissues and be detected by gamma cameras. Positron emitters are also employed, particularly in Positron Emission Tomography (PET) imaging, where the positron annihilates with an electron, producing two gamma photons that are detected simultaneously.

[0009] The radionuclide 64Cu is a well known positron emitting radionuclide with a half-life of 12.7 hours, and has been used for labeling imaging agents successfully. In addition to positron emission, 64Cu also decays by both beta minus particles and gamma radiation. Organ dosimetry calculations based on the human biodistribution of labelled compounds have, however, deemed 64Cu ineffective for therapeutic purposes. For this reason, 64Cu labelled compounds are used as imaging agents either alone for diagnostic purposes or combined with a 67Cu labelled counterpart for therapeutic purposes working together as a theranostic pair. One example may be 64Cu-SARTATE and 67Cu-SARTATE developed by Clarity pharmaceuticals.

[0010] In cancer disease, in particular solid tumors and metastasis, formation of new blood vessels is essential for the cancer to sustain growth. New blood vessels can be formed by two different mechanisms: vasculogenesis or angiogenesis. Angiogenesis is the formation of new blood vessels by branching from existing vessels. The primary stimulus for this process may be inadequate supply of nutrients and oxygen (hypoxia) to cells in a tissue. The cells may respond by secreting angiogenic factors, of which there are many; one example, which is frequently referred to, is vascular endothelial growth factor (VEGF). These factors initiate the secretion of proteolytic enzymes that break down the proteins of the basement membrane, as well as inhibitors that limit the action of these potentially harmful enzymes. The other prominent effect of angiogenic factors is to cause endothelial cells to migrate and divide. Endothelial cells that are attached to the basement membrane, which forms a continuous sheet around blood vessels on the contralumenal side, do not undergo mitosis. The combined effect of loss of attachment and signals from the receptors for angiogenic factors is to cause the endothelial cells to move, multiply, and rearrange themselves, and finally to synthesise a basement membrane around the new vessels.

[0011] Angiogenesis is prominent in the growth and remodelling of tissues, including wound healing and inflammatory processes. Tumors must initiate angiogenesis when they reach millimetre size in order to keep up their rate of growth. Angiogenesis is accompanied by characteristic changes in endothelial cells and their environment. The surface of these cells is remodeled in preparation for migration, and cryptic structures are exposed where the basement membrane is degraded, in addition to the variety of proteins which are involved in effecting and controlling proteolysis. In the case of tumors, the resulting network of blood vessels is usually disorganised, with the formation of sharp kinks and also arteriovenous shunts. Inhibition of angiogenesis is also considered to be a promising strategy for antitumor therapy.

[0012] As mentioned above, angiogenesis plays an important role in many pathological processes such as cancer. Integrin aVp3 is a cell adhesion molecule and is highly expressed on activated endothelial cells and tumor cells, but not on resting endothelial cells. During the past decade, many studies have confirmed the importance of integrin aVp3 as a specific target for neoangiogenesis, due to its role in the tumor growth and metastasis. Extracellular matrix (ECM) proteins such as vitronectin, fibrinogen and fibronectin interact with integrin aVp3 via the amino acid sequence Arg-Gly-Asp (RGD). Often reported RGD peptides used for radiolabeling are the cyclic pentapeptides cyclo(Arg-Gly-Asp-D-Phe-Lys), c(RGDfK) and cyclo(Arg-Gly-Asp-D- Tyr-Lys), c(RGDyK), which only differ with one amino acid.

[0013] OXBOEL J. et al. / NUCLEAR MEDICINE AND BIOLOGY 2014, Vol. 41 , Pages 259- 267) disclose the uptake of 68GA-NODAGA-E[c(RGDyK)]2 and 64Cu-NODAGA-E[c(RGDyK)]2 in tumors and the tumors being visible on PET images. The paper further mentions that angiogenesis plays an important role in cancer. Meanwhile, there is no indication nor evidence presented in the paper indicating that [64Cu]NODAGA-E[c(RGDyK)]2, nor that 64Cu as such, may provide any therapeutic effect in cancer patients.

[0014] SUMMARY OF THE INVENTION

[0015] It was surprisingly found that the radionuclide 64Cu well known for its imaging properties provides therapeutic efficacy at doses far below the theoretical dose, thereby enabling therapeutic application of 64Cu labeled conjugates according to the present invention in human patients suffering from cancer.

[0016] In a first aspect, the present invention relates to a 64Cu labelled conjugate for use in treatment of cancer, wherein said 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0017] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0018] In a preferred embodiment of the first aspect, said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

[0019] In a second aspect, the present invention relates to a method of treatment of a cancer disease by administering to a patient a 64Cu labelled conjugate, wherein sad 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0020] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0021] In a preferred embodiment of the second aspect, said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

[0022] In a third aspect, the present invention relates to a pharmaceutical formulation comprising a 64Cu labelled conjugate, wherein said 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0023] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0024] The 64Cu labelled conjugates of the present invention offer a new palette of therapeutic compounds for cancer treatment. In the present invention, 64Cu shown therapeutic efficacy at radiation doses providing much lower gray (Gy) than what is generally found necessary for curative treatment, thereby providing a much wider therapeutic window in which the dose providing the optimal balance of therapeutic effect and side effects can be achieved compared to inferior counterparts.

[0025] For the purpose of completeness, it is herewith clarified that the 64Cu labelled conjugates according to the invention and all the embodiments are preferably consisting of the specified components. That is, in a preferred form, the first aspect of the present invention relates to: a 64Cu labelled conjugate for use in treatment of cancer, wherein said 64Cu labelled conjugate consists of: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0026] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0027] In a preferred form, the second aspect of the present invention relates to: a method of treatment of a cancer disease by administering to a patient a 64Cu labelled conjugate, wherein sad 64Cu labelled conjugate consists of: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0028] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0029] In a preferred form, the third aspect of the present invention relates to: a pharmaceutical formulation comprising a 64Cu labelled conjugate, wherein sad 64Cu labelled conjugate consists of: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0030] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0031] BRIEF DESCRIPTION OF THE FIGURES

[0032] Figure 1 : A) shows a representative PET / CT images of the biodistribution of 64Cu-DOTA- AE105 in mice with xenograft tumors (U87MG). Images are taken 0.5H, 2H, and 24H after injection of 10 MBq Cu-64-DOTA-AE105. For each timepoint, the left image represents an axial slice intersecting the center of the tumor. The right image represents the Ml P image. B) shows In vivo biodistribution 0.5H, 2H and 24H post-dosing of 10 MBq Cu-64-DOTA-AE105, estimated by quantitative region interest (ROI) analysis. Graph show %ID / g for all listed organs. N=2-3. Bars represent mean ± SEM. C) shows Ex vivo biodistribution of Cu-64- DOTA-AE10524H post-dosing of 10 MBq Cu-64DOTA-AE105. Bars represent mean ± SEM. N=3.

[0033] Figure 2: A) shows mean tumor growth for up to 3 weeks after administration of a single dose of 130 MBq of 64Cu-DOTA-AE105 as therapy. Dose was administered at day 0. Carry forward was applied for mean tumor growth curve until <60% animals were still alive in the groups. The vertical dotted lines indicate the initiation of treatment with Cu-64-DOTA-AE105. Bars represent mean ± SEM. N=5 / group. B) shows tumor volume comparison at study day 13 (the last day where 60% or more animals were alive within the two groups). The effect of Cu-64-DOTA-AE105 was tested against Vehicle. A significant effect was observed following treatment with Cu-64-DOTA-AE105 compared to Vehicle (Unpaired t-test, p=0.0139). Bars represent mean ± SEM. N=5 / group. C) shows animal survival for up to 27 days after treatment initiation. Each event represents an animal euthanized to humane endpoints. Animals euthanized due to tumor wounds are censured in the survival curve. A significant survival was observed following treatment with Cu-64-DOTA-AE105 compared to Vehicle (Mantel Cox’s test, P= 0.0164). N= 5 / group

[0034] Figure 3: A) shows treatment efficacy of 67Cu-DOTA-AE105 in a human glioblastoma xenograft mouse model. B) shows Kaplan-Meier curves of survival data from the treatment efficacy study of 67Cu-DOTA-AE105 in a human glioblastoma xenograft mouse model.

[0035] Figure 4: A) shows mean tumor growth of group A-E for up to 5 weeks after therapy initiation. Carry forward was applied for mean tumor growth curve until <60% animals were still alive in the groups. The vertical dotted lines indicate the initiation of treatment with Cu-64-DOTA- AE105. Bars represent mean ± SEM. N=5 / group.B) shows tumor volume comparison at study day 17 (the last day where 60% or more animals were alive within the all groups). The effect of a single dose of 19, 37 and 111 MBq Cu-64-DOTA-AE105 (Group B-D) and the effect of fractionated dosing of 3 times 37 MBq Cu-64-DOTA-AE105 (Group E) were tested against Vehicle (Group A) by One-way ANOVA with Dunnett’s multiple comparison. A significant effect was observed following treatment with 111 MBq Cu-64-DOTA-AE105 compared to Vehicle and 3 times 37 MBq Cu-64-DOTA-AE105 as fractionated dosing. Bars represent mean ± SEM. N=3-5 / group.

[0036] Figure 5: A) to E) shows animal survival for group A-E for up to 5 weeks after treatment initiation. Each event represents an animal euthanized to humane endpoints. Animals euthanized due to tumor wounds are censured in the survival curve. N= 5 / group.

[0037] Figure 6: shows mean tumor growth for up to 8 weeks after administration of a single dose of 150MBq of 64Cu-DOTA-AE105 in a NCI-H1993 tumor model. Carry forward was applied for mean tumor growth curve until <60% animals were still alive in the groups. The vertical dotted lines indicate the initiation of treatment with Cu-64-DOTA-AE105. Bars represent mean ± SEM. N=5 / group.

[0038] Figure 7: A) - C) shows treatment efficacy of 64Cu-NODAGA-E[c(RGDyK)]2in a human glioblastoma xenograft mouse model. D) shows Kaplan-Meier curves of survival data from the treatment efficacy study of 64Cu-NODAGA-E[c(RGDyK)]2in a human glioblastoma xenograft mouse model.

[0039] DETAILED DESCRIPTION OF THE INVENTION

[0040] It was surprisingly found that the radionuclide 64Cu, well known for its imaging properties, provides therapeutic efficacy at doses far below the theoretical dose thereby enabling therapeutic application of 64Cu labelled conjugates according to the present invention in human patients suffering from cancers.

[0041] Becquerel (Bq) is the unit of radioactivity in the International System of Units (SI). One becquerel is defined as an activity of one decay per second. Bq is a measure of activity dose (what is administered). In context of human dosing the commonly used multiples M and G are used, i.e. MBq (megabecquerel, 106Bq) GBq (gigabecquerel, 109Bq). Gray (Gy) is the unit of ionizing radiation dose in the International System of Units (SI), defined as the absorption of one joule of radiation energy per kilogram of matter. Thus, Gy is a measure of deposited dose in tissue.

[0042] OLINDA / EXM is the most widely used and FDA approved software for standardized dose calculations for diagnostic and therapeutic radiopharmaceuticals (Stabin MG, Sparks RB, Crowe E. OLINDA / EXM: the second-generation personal computer software for internal dose assessment in nuclear medicine. J Nucl Med. 2005 Jun;46(6):1023-7. PMID: 15937315.

[0043] Stabin MG et al. 2005, and FDA.gov 2004 FDA approval of Olinda Exm (https: / / www.accessdata.fda.gov / cdrh docs / pdf3 / k033960.pdf)). The program calculates the absorbed radiation dose to various organs according to the MIRD (Medical Internal Radiation Dose Committee of the Society of Nuclear Medicine) technique.

[0044] Based on the human biodistribution of the imaging ligand 64Cu-DOTA-AE105, the inventor calculated the organ dosimetry to be anticipated from 67Cu-DOTA-AE105 (table 1), using the standard software OLINDA / EXM.

[0045] Table 1: Organ doses of 67Cu-DOTA-AE105 are given in mGy / MBq. Effective dose is given in units of mSv / MBq.

[0046] Combining these calculations with the generally accepted maximum allowed radiation doses to organs (Wahl (Wahl et al., 2021) and Emami (Emami, 2013) and table 2, central column, maximal allowed administered doses of 67Cu-DOTA-AE105 for each organ could be calculated (table 2, right column) and the dose limiting organ was red bone marrow with a limit of administered radioactivity dose of 24.2 GBq 67Cu-DOTA-AE105.

[0047] Table 2: Calculated values for maximum allowed activity of 67Cu-DOTA-AE105. Values are based on dose limits from Wahl et al. 2021 and Emami et al. 2015. Assuming a tumor uptake of a Standardized Uptake Value (SUV) of 10, which is a relevant level for tumors in general with our compound (Persson et al 2015) would result in a radiation dose to a 10 g tumor of 26.1 Gy (for dependence of tumor size, please see table 3). Table 3: Calculated total tumor dose at maximum administered activity, as a function of spherical tumor size.

[0048] Generally accepted radiation doses needed for palliative radiation therapy are >30 Gy and for curative radiation therapy >50 Gy. Accordingly, the target dose to tumors cannot be achieved before reaching the dose limit of healthy organs.

[0049] When the same calculations are made for the imaging ligand 64Cu-DOTA-AE105, a maximal administered dose of 112 GBq of 64Cu-DOTA-AE105 was calculated to be the highest dose before reaching the maximum dose in the dose-limiting organ (red bone marrow). At this dose, the radiation dose delivered to a 10g tumor with an uptake of SUV=10, would then be 20.8 Gy leading to the same conclusion that the target doses of >30 Gy (palliative treatment) or >50 Gy (curative treatment) could not be achieved before reaching the dose limit of healthy organs. For details, please see tables 4, 5, and 6. Table 4: Organ doses of 64Cu-DOTA-AE105 are given in mGy / MBq. Effective dose is given in units of mSv / MBq.

[0050] Table 5: Calculated values for maximum allowed activity of 64Cu-DOTA-AE105. Values are based on dose limits from Wahl et al 2021 and Emami et al. 2015.

[0051] Table 6: Calculated total tumor dose at maximum administered activity, as a function of spherical tumor size. From the calculations using OLINDA / EXM software, it can also be concluded that a 5-6 fold higher administered activity dose of 64Cu-DOTA-AE105 compared to 67Cu-DOTA-AE105 is needed to obtain the same radiation dose to a tumor (112 GBq / 20.8 Gy = 5.4 GBq / Gy for 64Cu-DOTA-AE105 vs. 24.2 GBq / 26.1 Gy = 0.93 GBq / Gy for 67Cu-DOTA-AE105). However, surprisingly, the inventor found in a series of experiments that this was not the case. Accordingly, in comparative tumor efficacy studies using either 67Cu-DOTA-AE105 or 64Cu- DOTA-AE105, it was found that a dose of 111 MBq of 64Cu-DOTA-AE105, i.e. less than 2- fold higher, was as effective as a dose of 60 MBq of 67Cu-DOTA-AE105 indicating that in clear contrast to the delivered doses, 64Cu-DOTA-AE105 was surprisingly effective in treating human xenograft tumors (see example 4 and 5). These studies (Example 4 and 5) were conducted in mice bearing human xenograft tumors, thus the therapeutic efficacy at surprisingly low radiation dose observed for 64Cu-DOTA-AE105 is directly relevant for human therapy. This is ground-breaking as this allows for an approximately 2 14 - 3-fold reduction in delivered dose in Gy when using 64Cu-DOTA-AE105 vs. 67Cu-DOTA-AE105. In other words, the calculated dose in the human dosimetry studies, where the maximal dose with respect to organs of 112 GBq lead to a radiation dose of 20.8 Gy which was deemed ineffective according to standard limits for target tumors doses of >30 Gy (palliative) or >50 Gy (curative), suddenly becomes relevant if the efficacy is 2 14-3 fold more effective than the similar dose in Gy delivered by 67Cu-DOTA-AE105, i.e. the efficacy to be expected in humans of 20.8 Gy would be equivalent to >50 Gy (52-62) Gy, which would be highly relevant for clinical use.

[0052] Furthermore, the inventor has observed therapeutic effects of other 64 labelled compounds including 64Cu-NODAGA-E[c(RGDyK)]2(cf. Example 9) and 64Cu-DOTA-TATE following administering a dose of 111 MBq. Thus, the advantages described in detail above exemplified by 64Cu-DOTA-AE105 are generally applicable to 64Cu labelled conjugates according to the invention, in particular conjugates having a peptide based targeting moiety.

[0053] Clinical studies with67Cu-SARTATE (Clinical trial NCT04438304) Phase I dose escalation studies reported single doses between 4.5 and 22 GBq (approx. 120-600 mCi) per cycle. Up to 4 treatment cycles have been reported in some protocols. Thus, the cumulative activity dose is typically in the range of -18-88 GBq (approx. 0.5-2.4 Ci), in round numbers 20-100 GBq depending on response and tolerance. It would normally be expected that 5-6 times higher activity doses of the 64Cu version should be used (i.e. 110-500 GBq). Therefore, it was very surprising to find here that less than or equal to a factor of 2 (40-200 GBq) is sufficient.

[0054] Targeting moiety

[0055] The targeting moiety in the context of the present invention may be any entity providing targeting of 64Cu to a desired destination (target) within the human body, such as but not limited to antibodies, peptides, and small molecules, preferably said targeting is selective towards the intended target whereby off target exposure to radiation is limited as much as possible.

[0056] One example of suitable targeting moieties according to the present invention is antibodies (polyclonal antibodies, monoclonal antibodies) and antibody derived targeting moieties such as, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody preserving their functional potential while being more limited in size.

[0057] In the context of large targeting moiety such as antibodies, the targeting moiety provides the possibility for attaching more than one chelating agent, thereby increasing the radionuclide load per conjugate. Thus, 64Cu labelled conjugates according to all aspects of the invention comprising one or more chelating agents, preferably two or more chelating agents, are preferred when the targeting moiety is an antibody.

[0058] In one embodiment of all aspects of the invention, the targeting moiety is selected from the group comprising antibodies (polyclonal antibodies or monoclonal antibodies), antibody derived targeting moieties such as, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody.

[0059] Another example of suitable targeting moieties according to the present invention is antibody mimetics, such as affibody molecules.

[0060] In one embodiment of all aspects of the invention, the targeting moiety is an affibody molecule.

[0061] Another example of suitable targeting moieties according to the present invention is nucleic acid molecules, such as aptamers.

[0062] In one embodiment of all aspects of the invention, the targeting moiety is nucleic acid molecules, preferably aptamers.

[0063] Another example of suitable targeting moieties according to the present invention is small molecules.

[0064] In one embodiment of all aspects of the invention, the targeting moiety is a small molecule. Another example of suitable targeting moieties is peptides or peptidomimetics which has shown promising potential for specific targeting in radionuclide based therapy. Peptides are generally considered by the skilled person to be chains of 2 to 50 amino acids linked by peptide bonds. Peptides may be linear, branched or cyclic molecules build from 2 to 50 amino acids and may have post-translational modifications.

[0065] In a preferred embodiment of all aspects of the invention, the targeting moiety is a peptide or peptidomimetics.

[0066] In a more preferred embodiment of all aspects of the invention, the targeting moiety is a peptide composed of 2 to 50 amino acids, preferably 2 to 20 amino acids.

[0067] The choice of specific targeting moiety will depend on the target being selected for therapy and its location within the human body. Targeting moieties according to all aspects of the invention are suitable for coupling with a chelating agent according to all aspects of the invention thereby enabling labelling with the radionuclide 64Cu.

[0068] Preferred targeting moieties according to all aspects of the present invention are RGD based targeting moieties providing targeting of integrin avp3. Several peptide-based targeting moieties have been developed based on the triple amino acid sequence Arg-Gly-Asp (RGD) that specifically binds to integrin avp3. Several of these peptides have been used as tracers for PET imaging, many of those will also provide good targeting moieties for 64Cu labelled conjugates according to all aspects of the present invention providing selective targeting of integrin OvPsand stable retention at the target for long enough such that 64Cu may provide the therapeutically effective radiation dose locally at site of the targeted cancer disease.

[0069] In a preferred embodiment of all aspects of the invention, the targeting moiety is an RGD based integrin QvPs targeting peptide, preferably E[c(RGDyK)]2.

[0070] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin QvPs targeting peptide and a chelating agent suitable for binding radiometals wherein the RGD based integrin QvPs targeting peptide is coupled to 64Cu by the chelating agent. In a more preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2. and a chelating agent suitable for binding radiometals; wherein the E[c(RGDyK)]2. targeting peptide is coupled to 64Cu by the chelating agent.

[0071] In a most preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate is 64Cu-NODAGA-E[c(RGDyK)]2.

[0072] The compound 64Cu-NODAGA-E[c(RGDyK)]2 is produced by complexing the positron emitting radionuclide, Copper-64, with a NODAGA chelator attached to the peptide Glutamoyl-bis- cyclo(L-argininylglycyl-L-a-aspartyl-D-tyrosyl-L-lysyl). The chemical structure of 64Cu- NODAGA-E[c(RGDyK)]2 is shown below.

[0073] Chelator

[0074] The use of various chelating agents suitable for binding radiometals for radio labeling of targeting moieties, such as peptides, is well known in the art. Suitable chelating agents generally include those which contain a tetradentate ligand with at least one sulfur group available for binding the metal radionuclide, such as the known N3S and N2S2 ligands. The chelating agent is coupled to the targeting moiety by standard methodology known in the field of the invention and may be added at any location on the targeting moiety, provided that the biological activity such as binding properties and specificity of the targeting moiety is not adversely affected. In context of peptide-based targeting moieties, the chelating group is preferably covalently coupled to the amino terminal amino acid of the peptide. The chelating group may advantageously be attached to the peptide during solid phase peptide synthesis or added by solution phase chemistry after the peptide has been obtained. Preferred chelating agents include DOTA (1 ,4,7, 10-tetrakis(carboxymethyl)-1 ,4,7,10 tetraazacyclo dodecane), DOT AM (2-[4, 7, 10-tris(2-amino-2-oxoethyl)- 1 ,4, 7, 10-tetrazacyclododec- 1 -yl]acetam ide) ,

[0075] NOTA (2,2',2”-(1 ,4,7-triazacyclononane-1 ,4,7-triyl)triacetic acid), NODAGA, SARCOPHAGINE (3,6,10,13,16,19-hexaazabicyclo(6,6,6)icosane), CB-TE2A (1 ,4,8,11- Tetraazabicyclo[6.6.2]hexadecane-4, 11 -diacetic acid) and derivatives thereof, which constitute an important class of chelators for biomedical applications as they accommodate very stably a variety of di- and trivalent metal ions.

[0076] DOTA (Dodecane tetraacetic acid) with the chemical formula (1 ,4,7, 10- tetrakis(carboxymethyl)-1 ,4,7,10 tetraazacyclo dodecane) and its derivatives, are particularly acknowledged to constitute an important class of chelators for biomedical applications as they accommodate very stably a variety of di- and trivalent metal ions, including 64Cu.

[0077] DOTAM also known as TCMC is a DOTA analogue.

[0078] SARCOPHAGINE (Sar) is a bicyclic cage-like chelator molecule derived from cyclam. The chemical formula of sarcophagine is 3,6,10,13,16,19-hexaazabicyclo(6,6,6)icosane and additional functional or non-functional groups are often linked to this structure, thereby creating derivatives of sarcophagine, such as in DiAmSar (1 ,8-diamino-Sar), AmBaSar (4-((8-amino- 3,6,10,13,16,19-hexaazabicyclo [6.6.6] icosane-1-ylamino)methyl)benzoic acid), and MeCOSar (5-(8-methyl-3,6, 10,13,16,19-hexaaza-bicyclo[6.6.6]icosan- 1 -ylamino)-5- oxopentanoic acid). Sarcophagine and derivatives thereof are suitable chelating agents for use in the 64Cu labelled conjugates of all aspects of the present invention and are preferred for 64Cu labelled conjugates.

[0079] In one embodiment of all aspects of the present invention, the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A and derivatives thereof, preferably DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB-TE2A. In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and a chelating agent selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A and derivatives thereof, preferably DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB-TE2A; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0080] In a more preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2.and the chelating agent selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A and derivatives thereof, preferably DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB-TE2A; wherein the E[c(RGDyK)]2. targeting peptide is coupled to 64Cu by the chelating agent.

[0081] In a preferred embodiment of all the aspects of the invention, the chelating agent is selected from the group consisting of NODAGA, DOTA, DOTAM, SARCOPHAGINE, and derivatives thereof, preferably NODAGA, DOTA, DOTAM, SARCOPHAGINE, DiAmSar, AmBaSar, or MeCOSar.

[0082] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and a chelating agent selected from the group consisting of NODAGA, DOTA, DOTAM, SARCOPHAGINE, and derivatives thereof, preferably DOTA, DOTAM, SARCOPHAGINE, DiAmSar, AmBaSar, or MeCOSar; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0083] In a more preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2.and the chelating agent selected from the group consisting of NODAGA, DOTA, DOTAM, SARCOPHAGINE, and derivatives thereof, preferably DOTA, DOTAM, SARCOPHAGINE, DiAmSar, AmBaSar, or MeCOSar; wherein the E[c(RGDyK)]2. targeting peptide is coupled to 64Cu by the chelating agent.

[0084] In a more preferred embodiment of all aspects of the present invention, the chelating agent is DOTA. In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and the chelating agent is DOTA; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0085] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is DOTA; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0086] In one embodiment of all aspects of the present invention, the chelating agent of 64Cu labelled conjugates is DOTAM.

[0087] In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin QvPa targeting peptide and the chelating agent is DOTAM; wherein the RGD based integrin QvPa targeting peptide is coupled to 64Cu by the chelating agent.

[0088] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is DOTAM; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0089] In one embodiment of all aspects of the present invention, the chelating agent of the 64Cu labelled conjugates is NOTA.

[0090] In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and the chelating agent is NOTA; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0091] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is NOTA; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0092] In one embodiment of all aspects of the present invention, the chelating agent of the 64Cu labelled conjugates is NODAGA. In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and the chelating agent is NODAGA; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0093] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is NODAGA; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0094] In one embodiment of all aspects of the present invention, the chelating agent of the 64Cu labelled conjugates is MeCOSar.

[0095] In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin QvPa targeting peptide and the chelating agent is MeCOSar; wherein the RGD based integrin QvPa targeting peptide is coupled to 64Cu by the chelating agent.

[0096] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is MeCOSar; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0097] In one embodiment of all aspects of the present invention, the chelating agent of the 64Cu labelled conjugates is CB-TE2A.

[0098] In one embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises an RGD based integrin avPs targeting peptide and the chelating agent is CB-TE2A; wherein the RGD based integrin avPs targeting peptide is coupled to 64Cu by the chelating agent.

[0099] In a preferred embodiment of all aspects of the invention, the 64Cu labelled conjugate comprises the targeting peptide E[c(RGDyK)]2and the chelating agent is CB-TE2A; wherein the targeting peptide E[c(RGDyK)]2is coupled to 64Cu by the chelating agent.

[0100] Diseases

[0101] The inventor has shown therapeutic efficacy of a 64Cu labelled conjugates, in two different human xenograft cancer models. It is generally accepted that therapeutic efficacy across two or more different cancer diseases constitutes a very strong support for therapeutic efficacy in general for cancers expressing the relevant target. Thus, 64Cu labelled conjugates according to the present invention are suitable for radionuclide-based therapy of human cancer. More specifically, 64Cu labelled conjugates targeting integrin avp3, such as RGD based 64Cu labelled conjugates of the present invention, including 64Cu-NODAGA-E[c(RGDyK)]2, allows for treatment of cancers sensitive to disruption of angiogenesis.

[0102] In general, angiogenesis is essential for all solid cancers. In accordance with the present invention, cancers sensitive to disruption of angiogenesis are e. g. breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, thyroid cancer, head and neck cancer, cervical cancer, bladder cancer, astrocytomas, choriocarcinomas, glioblastomas, gliomas, hemangiomas (childhood, capillary), hepatomas, hyperplastic endometrium, Kaposi sarcoma, melanoma, neuroblastomas, seminomas.

[0103] In one embodiment of all aspects of the present invention, the cancer is selected from the group consisting of breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, thyroid cancer, head and neck cancer, cervical cancer, bladder cancer, astrocytomas, choriocarcinomas, glioblastomas, gliomas, hemangiomas (childhood, capillary), hepatomas, hyperplastic endometrium, Kaposi sarcoma, melanoma, neuroblastomas, seminomas. Preferably breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, head and neck cancer, cervical cancer, glioblastomas, gliomas, melanoma, neuroblastomas.

[0104] In a preferred embodiment of all aspects of the invention, the cancer is selected from the group consisting of high-grade glioma, breast cancer, melanoma, colorectal cancer, pancreatic cancer, neuroendocrine cancer.

[0105] In a more preferred embodiment of all aspects of the invention, the cancer is selected from the group consisting of high-grade glioma, colorectal cancer, pancreatic cancer, neuroendocrine cancer.

[0106] In a most preferred embodiment of all aspects of the invention, the cancer is high-grade glioma.

[0107] Dosage and route of administration 64Cu labelled conjugates according to the present invention may be administered systemically or locally to a patient, preferably a human patient.

[0108] Systemic administration of 64Cu labelled conjugates according to all aspects of the present invention are typically administered intravenously and may be administered intravenously in any conventional medium for intravenous injection.

[0109] Suitable cumulative doses of 64Cu labelled conjugates according to the present invention for treatment of cancer in human patients are in the range of 40 GBq to 200 GBq, such as 40 GBq to 163 GBq, preferably 50 GBq to 112 GBq whereby a good therapeutic effect is achieved while minimizing side effects.

[0110] In one embodiment of all aspects of the present invention, the cumulative dose administered intravenously to a human patient of the 64Cu labelled conjugates according to any of the previous embodiments is in the range of 40 GBq to 200 GBq, such as 40 GBq to 163 GBq, preferably 50 GBq to 112 GBq.

[0111] In some cancer diseases, systemic exposure to radionuclide-based therapy e.g. by intravenous administration does not provide optimal targeting of the cancer and localized administration may provide for better targeting of the cancer tissue. Localized administration may be provided by direct injection into the tumor (intralesional), installation into a hollow organ (intracavitary), e.g. the urinary bladder (intravesicular or intravesical), or by using a catheter or other suitable tubing providing access to the relevant site of administration or administration into the blood stream a site with delivery more directly to the tumor such as Intra-arterial administration where the drug is put right into the main artery that supplies blood to the tumor. Routes for localized administration are known by the skilled person and further examples of intracavitary administration where the drug may be given through a catheter into an enclosed area of the body are intraperitoneal where the drug is administered to the abdomen or belly, or intrapleural where the drug is administered to the chest.

[0112] A specific example is administration of a dose of 64Cu labelled conjugates to the urinary bladder, by bladder installation, whereby a suitable volume of liquid comprising a single dose of approximately 2GBq to 10GBq of the 64Cu labelled conjugates is delivered to the bladder lumen and maintained there for a period of time, such as 10 minutes to 60 minutes before draining the bladder. The administration is repeated a suitable number of times, such as 5 to 35 doses in order to reach a desired cumulative dose. Suitable cumulative doses for localized treatment of the bladder are 335GBq installed for 1 h or 1 ,340 GBq installed for 15 min suitable ranges of cumulative doses for localized treatment of the bladder are, up to 335GBq installed for 15 min to 2 h, such as for 15min to 1 h or up to 1 ,340GBq if installed for up to 15 min, such as 5 min to 15 min, preferably in the range of 50-335GBq installed for 15 min to 2 h, such as for 15 min to 1 or 200-1 , 340GBq if installed for up to 15 min, such as 5 min to 15 min, most preferred in the range of 100-335GBq installed for 15 min to 2 h, such as for 15 min to 1 h or 400-1 , 340GBq if installed for up to 15 min, such as 5 min to 15 min.

[0113] In one embodiment of all aspects of the present invention, the cumulative dose administered locally to the bladder of a human patient of the 64Cu labelled conjugate according to any of the previous embodiments is up to 335GBq installed for 15 min to 2 h, such as for 15min to 1 h or up to 1 ,340GBq if installed for up to 15 min, such as 5 min to 15 min, preferably in the range of 50-335GBq installed for 15 min to 2 h, such as for 15min to 1 or 200- 1 ,340GBq if installed for up to 15 min, such as 5 min to 15 min, most preferred in the range of 100-335GBq installed for 15 min to 2 h, such as for 15 min to 1 h or 400-1 , 340GBq if installed for up to 15 min, such as 5 min to 15 min.

[0114] Preferably, single doses of 2 GBq to 20 GBq are administered to the bladder lumen, and the administration is repeated 5 to 35 times until the desired cumulative dose is achieved.

[0115] Another specific example is administration of a dose of 64Cu labelled conjugate according to the present invention locally to the brain. Local administration to the brain may be provided by use of suitable catheter systems where the placement of the catheter is obtained through stereotactic technique, and whereby a suitable volume of liquid comprising the 64Cu labelled conjugate is delivered locally to the brain. When combined with pressure infusion, the technique is also known as convection-enhanced diffusion.

[0116] The local administration may also be achieved by intra-arterial administration to the brain, also known as super-selective intra-arterial cerebral infusion (SSIACI) where delivery is through a targeted vascular territory. In addition, these techniques may be combined with blood-brain barrier disruption such as mannitol or Focused Ultrasound.

[0117] Embodiments 1. Embodiment 1 , A 64Cu labelled conjugate for use in treatment of cancer, wherein said 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0118] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0119] 2. Embodiment 2, The 64Cu labelled conjugate for use according to embodiment 1 , wherein said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

[0120] 3. Embodiment 3, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 2, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A, or derivatives thereof.

[0121] 4. Embodiment 4, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 3, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB- TE2A .

[0122] 5. Embodiment 5, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 4, wherein the chelating agent is NODAGA.

[0123] 6. Embodiment 6, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 5, wherein the cumulative dose is administered systemically to a human patient, preferably by the intravenous route, in the range of 40 GBq to 200 GBq, such as 40GBq to 163GBq, preferably 50GBq to 112GBq.

[0124] 7. Embodiment 7, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 5, wherein the 64Cu labelled conjugate is administered locally.

[0125] 8. Embodiment 8, The 64Cu labelled conjugate for use according to embodiment 7, wherein the 64Cu labelled conjugate is administered locally to the brain or bladder.

[0126] 9. Embodiment 9, The 64Cu labelled conjugate for use according to embodiment 8, wherein said 64Cu labelled conjugate is administered locally to the urinary bladder by bladder 1 installation in a cumulative dose of up to 335GBq installed for 15 min to 2 h or up to 1 ,340GBq installed for 15 min or less, preferably in the range of 50-335GBq installed for 15min to 2 h or 200-1 , 340GBq installed for 15 min or less, most preferred in the range of 100-335GBq installed for 15min to 2 h or 400-1 , 340GBq if installed for 15 min or less is administered locally to the urinary bladder. Embodiment 10, The 64Cu labelled conjugate for use according to any of the embodiments 8 and 9, wherein said 64Cu labelled conjugate is administered locally to the urinary bladder by bladder installation in a cumulative dose of up to 335GBq installed for 15 min to 1 h or up to 1 ,340GBq installed for 5 min to 15 min, preferably in the range of 50-335GBq installed for up to 1 h or 200-1 , 340GBq installed for 5 min to 15 min, most preferred in the range of 100-335GBq installed for up to 1 h or 400-1 , 340GBq if installed for 5 min to 15 min is administered locally to the urinary bladder by bladder installation. . Embodiment 11 , The 64Cu labelled conjugate for use according to any of the embodiments 9 to 10, wherein said cumulative dose is administered in single doses of 2 GBq to 10 GBq. Embodiment 12, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 11 , wherein the targeting moiety is selected from the group consisting of antibodies, antibody derived targeting moieties, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody, antibody mimetics, affibody molecules, nucleic acid molecules, aptamers, small molecules, peptides, peptidomimetics. Preferably the targeting moiety is selected from the group consisting of peptides and peptidomimetics, most preferably peptides. Embodiment 13, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 12, wherein the targeting moiety is an RGD based targeting moiety, preferably an RGD based integrin QvPa targeting peptide. Embodiment 14, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 13, wherein the targeting moiety is E[c(RGDyK)]2. Embodiment 15, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 14, wherein the 64Cu labelled conjugate has the formula:

[0127] 16. Embodiment 16, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 15, wherein said cancer is selected from the group consisting of breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, thyroid cancer, head and neck cancer, cervical cancer, bladder cancer, astrocytomas, choriocarcinomas, glioblastomas, gliomas, hemangiomas (childhood, capillary), hepatomas, hyperplastic endometrium, Kaposi sarcoma, melanoma, neuroblastomas, seminomas.

[0128] 17. Embodiment 17, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 16, wherein said cancer is selected from the group consisting of highgrade glioma, breast cancer, melanoma, colorectal cancer, pancreatic cancer, neuroendocrine cancer.

[0129] 18. Embodiment 18, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 17, wherein said cancer is selected from the group consisting highgrade glioma, colorectal cancer, pancreatic cancer, neuroendocrine cancer. 19. Embodiment 19, The 64Cu labelled conjugate for use according to any of the embodiments 1 to 18, wherein said cancer is high-grade glioma.

[0130] 20. Embodiment 20, A Method of treatment of a cancer disease by administering to a patient a 64Cu labelled conjugate, wherein sad 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64Cu

[0131] And wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0132] 21. Embodiment 21 , The method of treatment according to embodiment 20, wherein said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

[0133] 22. Embodiment 22, The method of treatment according to any of the embodiments 20 to 21 , wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A, or derivatives thereof.

[0134] 23. Embodiment 23, The method of treatment according to any of the embodiments 20 to 22, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB-TE2A .

[0135] 24. Embodiment 24, The method of treatment according to any of the embodiments 20 to 23, wherein the chelating agent is NODAGA.

[0136] 25. Embodiment 25, The method of treatment according to any of the embodiments 20 to 24, wherein said 64Cu labelled conjugate is administered systemically to a human patient, preferably by the intravenous route, in a cumulative dose of 40 GBq to 200 GBq, such as 40GBq to 163GBq, preferably 50GBq to 112GBq.

[0137] 26. Embodiment 26, The method of treatment according to any of the embodiments 20 to 24, wherein the 64Cu labelled conjugate is administered locally.

[0138] 27. Embodiment 27, The method of treatment according to embodiment 26, wherein the 64Cu labelled conjugate is administered locally to the brain or bladder. 28. Embodiment 28, The method of treatment according to embodiment 27, wherein said 64Cu labelled conjugate is administered locally to the urinary bladder by bladder installation in a cumulative dose of up to 335GBq installed for 15 min to 2 h or up to 1 ,340GBq installed for 15 min or less, preferably in the range of 50-335GBq installed for 15min to 2 h or 200-1 , 340GBq installed for 15 min or less, most preferred in the range of 100-335GBq installed for 15min to 2 h or 400-1 , 340GBq if installed for 15 min or less is administered locally to the urinary bladder.

[0139] 29. Embodiment 29, The method of treatment according to any of the embodiments 27 to 28, wherein said 64Cu labelled conjugate is administered locally to the urinary bladder by bladder installation in a cumulative dose of up to 335GBq installed for 15min to 1 h or up to 1 ,340GBq installed for 5 min to 15 min, preferably in the range of 50-335GBq installed for up to 1 h or 200-1 , 340GBq installed for 5 min to 15 min, most preferred in the range of 100-335GBq installed for up to 1 h or 400-1 , 340GBq if installed for 5 min to 15 min is administered locally to the urinary bladder by bladder installation. .

[0140] 30. Embodiment 30, The method of treatment according to any of the embodiments 28 to 29, wherein said cumulative dose is administered in single doses of 2 GBq to 10 GBq.

[0141] 31 . Embodiment 31 , The method of treatment according to any of the embodiments 20 to 30, wherein the targeting moiety is selected from the group consisting of antibodies, antibody derived targeting moieties, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody, antibody mimetics, affibody molecules, nucleic acid molecules, aptamers, small molecules, peptides, peptidomimetics. Preferably the targeting moiety is selected from the group consisting of peptides and peptidomimetics, most preferably peptides.

[0142] 32. Embodiment 32, The method of treatment according to any of the embodiments 20 to 31 , wherein the targeting moiety is an RGD based targeting moiety, preferably an RGD based integrin QvPa targeting peptide.

[0143] 33. Embodiment 33, The method of treatment according to any of the embodiments 20 to 32, wherein the targeting moiety is E[c(RGDyK)]2.

[0144] 34. Embodiment 34, The method of treatment according to any of the embodiments 20 to 33, wherein the 64Cu labelled conjugate has the formula: Embodiment 35, The method of treatment according to any of the embodiments 20 to 34, wherein said cancer is selected from the group consisting of breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, thyroid cancer, head and neck cancer, cervical cancer, bladder cancer, astrocytomas, choriocarcinomas, glioblastomas, gliomas, hemangiomas (childhood, capillary), hepatomas, hyperplastic endometrium, Kaposi sarcoma, melanoma, neuroblastomas, seminomas. Embodiment 36, The method of treatment according to any of the embodiments 20 to 35, wherein said cancer is selected from the group consisting of high-grade glioma, breast cancer, melanoma, colorectal cancer, pancreatic cancer, neuroendocrine cancer. Embodiment 37, The method of treatment according to any of the embodiments 20 to 36, wherein said cancer is selected from the group consisting high-grade glioma, colorectal cancer, pancreatic cancer, neuroendocrine cancer. 38. Embodiment 38, The method of treatment according to any of the embodiments 20 to 37, wherein said cancer is high-grade glioma.

[0145] 39. A pharmaceutical formulation comprising a) a targeting moiety b) a chelating agent suitable for binding radiometals c) radionuclide 64Cu wherein the targeting moiety is coupled to 64Cu by the chelating agent.

[0146] EXAMPLES

[0147] Example 1 : Radiolabeling

[0148] 64Cu labelling of DOTA-AE105 conjugate

[0149] A radionuclide labelled uPAR binding peptide conjugate according to the present invention was prepared by 64Cu labelling of DOTA-AE105 conjugate.

[0150] The compound was labeled with suitable molar activity e.g. 25 MBq / nmol and 322.4 MBq / nmol to generate stock solutions from which relevant doses such as 10 MBq 19 MBq, 30 MBq, 37 MBq, 60 MBq, 111 MBq and 130 MBq could be intravenously administered.

[0151] Compound(s) was labeled with a specific activity of 10 MBq / nmol (Biodistribution studies, Cu- 64), 322.4 MBq / nmol (Efficacy studies Cu-64).

[0152] When labeling with Cu-64, the dried Cu-64 was dissolved in Trace select water to an activity concentration between 2-30 GBq / mL.

[0153] • The desired activity was transferred to an Eppendorf tube, and the activity was measured.

[0154] • Labeling buffer (for low specific activity:0.25 M NaOAc, for High specific activity: 3 M NaOAc w. 50 mg / mL ascorbic acid) was added in ratio 1 :1 or to obtain an reaction volume between 80-200.

[0155] • The pH was measured by indicator paper. Estimated pH 5.0-5.5.

[0156] • The amount of compound was added to reach the desired molar activity.

[0157] • Reaction mixture was stirred at 80°C, 600 RPM for up to 15 min.

[0158] • The product may be purified depending on the degree of incorporation. • The final product was formulated in formulation buffer (20 mg / mL ascorbic acid, 1 mg / mL Tween-20, 5% EtOH in Sterile NaCI for injection) to the desired activity concentration and compounds concentration.

[0159] • The radiochemical purity (RCP) of the final product (EOS) was analyzed by radio-HPLC and radio-TLC.

[0160] Example 2 Biodistribution of 64Cu-DOTA-AE105

[0161] The biodistribution of 64Cu-DOTA-AE105 was evaluated in female NMRI nude mice bearing subcutaneous U87 MG tumors.

[0162] Three mice were included for in vivo and ex vivo biodistribution (Group A) on study day -1. The mean (±SEM) tumor volume at inclusion was 245.3 ± 48.5 mm3(range: 181.1-340.3 mm3) and the mean (±SEM) inclusion weight was 27.9 ± 1.4 g (range: 26.4- 30.6 g).

[0163] Each mouse received a single dose of 10 MBq 64Cu-DOTA-AE105 and underwent PET / CT imaging 0.5H, 2H and 24h post-dosing, ex vivo biodistribution was evaluated after the last imaging at the 24H time point.

[0164] Representative axial and maximum intensity projection (MIP) images with scale bar 0-20 %l D / g of one animal from group A are shown in Figure 1A.

[0165] The results of in vivo biodistribution of 10 MBq Cu-64-DOTA-AE105, is showed in figure 1 B. From the figure it can be appreciated that tumor uptake increased within 24 hours from 3.2 %l D / g at 0.5 hours to 5.4 %l D / g after 24 hours. Conversely, it was observed that the liver uptake decreased over time from 12.9 %l D / g at 0.5 hours to 6.3 %l D / g after 24 hours.

[0166] Results from ex vivo biodistribution is shown in figure 1C. From figure 1C it can be appreciated that 24 hours post-dosing, the liver displayed the highest ex vivo uptake, with a mean uptake of 7.4 %l D / g, while the tumor exhibited the second-highest uptake, with a mean of 6.7 %l D / g.

[0167] Example 3 Therapeutic effect of 64Cu-DOTA-AE105 in glioblastoma bearing xenograft mice

[0168] The treatment efficacy of 64Cu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous U87 MG tumors. The U87 MG tumor cell line is a human glioblastoma cell line and tumors were established subcutaneously in the flank region of the female NMRI nude mice. Ten animals were included and stratified into two groups based on tumor volume and body weight (n=5 / group) on study day -1.

[0169] The mean (±SEM) tumor volume at inclusion was 76.5 ± 8.2 mm3(range: 50.0-131.0 mm3) and the mean (±SEM) inclusion weight was 30.9 ± 0.7 g (range: 27.9- 33.7 g). No significant difference was observed for tumor volume or body weight between the groups (Student's t- test, p=0.94).

[0170] Animals received either vehicle (untreated control) or 130 MBq Cu-64-DOTA-AE105 at study day 0.

[0171] All mice were pre-treated with diet-gel five days prior to dosing and 5 days post dosing to prevent significant weight loss. Tumors and body weights were measured twice weekly for the duration of the study. Animals were taken down due to humane endpoint for tumor volume (> 1500mm3). The last observation for euthanized animals was carried forward until less than 60% of the animals were alive within the group.

[0172] The results are shown in figure 2A, from which it can be appreciated that a single dose of 130 MBq 64Cu-DOTA-AE105 significantly delayed tumor growth up to 17 days post dosing in the xenograft mice compared to vehicle control group.

[0173] Tumor volumes were compared between the two groups at the latest timepoint where 60% or more animals were alive within each group. The results are shown in figure 2B. Figure 2B shows tumor volume comparison between treatment with 64Cu-DOTA-AE105 and Vehicle control group at study day 13. The treatment with 130 MBq 64Cu-DOTA-AE105 led to a significant reduction in tumor growth and end tumor volume compared to animal receiving Vehicle (Unpaired t-test, p=0.0139). Additionally, relative animal body weight was unaffected on day 5 post-dosing.

[0174] Figure 2C show Kaplan-meier survival curves. It can be appreciated that the median survival for animals treated with Cu-64-DOTA-AE105 was 27 days, compared to 20 days for individuals in the Vehicle group, yielding a significant increase in survival for the treated group (Mantel Cox’s test, P= 0.0164). Example 4 Therapeutic effect of 67Cu-DOTA-AE105 in glioblastoma bearing xenograft mice

[0175] For benchmarking of the therapeutic efficacy of 64Cu-DOTA-AE105 in cancer therapy, the efficacy was compared to that of 67Cu labelled DOTA-AE105 conjugate (64Cu-DOTA- AE105). The results of example 3 was compared to the result of the following experiment.

[0176] The treatment efficacy of 67Cu-DOTA-AE105 was evaluated in female NMRI nude mice bearing subcutaneous U87 MG tumors. The U87 MG tumor cell line is a human glioblastoma cell line.

[0177] Animals were stratified into four groups (A, B, C and D) based on tumor volume and body weight on study day -2. The mean inclusion volume was 131.2 ± 8.6 mm3(range 66.0 - 258.3 mm3), while the inclusion weight was 29.5 ± 04 g (range 24.2 - 32.9 g). No significant difference in tumor volume or animal body weight was observed between the groups (oneway ANOVA, p>0.9 and p=0.3, respectively).

[0178] Animals were dosed with either vehicle (group A) or 67Cu-DOTA-AE105 (group B, C and D) at different activity levels on study day 0. Animals in group B received 29.6 ± 0.1 MBq, animals in group C received 29.6 ± 0.1 MBq, and animals in group D received 59.5 ± 0.2 MBq. At study day 14 animals in group C received an additional dosing with 30.1 ± 0.1 MBq. Data is presented as dosed measured - residual dose. No decay correction has been applied.

[0179] Tumor volume and body weight was monitored twice a week after the first dosing. The efficacy was evaluated over a period of 35 days. The mean tumor volumes for animals included in the study is presented in figure 3A. As can be observed in figure 3A, treatment with 60MBq 67Cu-DOTA-AE105 delayed tumor growth up to 13 days post dosing. Differences in tumor volume were evaluated at the latest timepoint where minimum 50% of animals were alive within each of the compared groups.

[0180] At study day 17, all treatment groups had significantly smaller tumor volumes when compared to the vehicle control group (Dunnett’s; A vs. B; p<0.03, A vs. C: p=0.02 and A vs. D: p<0.0001).

[0181] Animals were euthanized according to humane endpoints, which was reached when tumor sizes > 1500 mm3. Kaplan-Meier curves of survival data is presented in figure 3B. The median survival time for animals in the vehicle control group A was 17 days while animals in group B, C and D had a median survival time of 21 , 21 and 29 days, respectively.

[0182] The body weight did not differ between the groups during the study, indicating that the drug was safe and well tolerated.

[0183] 67Cu decays by P'(beta minus)-decay and has a half-life of 2.6 days. 67Cu is well known for therapeutic applications such as targeted radiotherapy. 64Cu on the contrary is well known as an imaging agent and not for therapy due to the limited P'(beta minus)-decay. Theoretically, providing 2,5 fold MBq of 64Cu would provide an equal dose of P'(beta minus)- irradiation as a given dose of 67Cu. Thus, figure 2 showing data from glioblastoma bearing xenograft mice treated with 130 MBq 64Cu-DOTA-AE105 theoretically equaling a dose of 52 MBq 67 Cu (130 divided by 2,5 = 52) is expected to show a similar or reduced therapeutic effect compared to the results shown in figure 3 showing data from glioblastoma bearing xenograft mice treated with 60MBq 67Cu-DOTA-AE105. Surprisingly, it can be appreciated that treatment with 64Cu-DOTA-AE105 provided a significantly delayed tumor growth up to 17 days post dosing as compared to 13 days observed for treatment with 60MBq 67Cu- DOTA-AE105 in the same tumor model. Thus, surprisingly the dose of 64Cu labelled DOTA- AE105 compound resulted in a better therapeutic effect than treatment with 60MBq 67Cu- DOTA-AE105.

[0184] Example 5 Dose-response of 64Cu-DOTA-AE105 in glioblastoma bearing xenograft mice

[0185] The dose dependent efficacy of 64Cu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous U87 MG tumors (see figure 4). The U87 MG tumor cell line is a human glioblastoma cell line.

[0186] Animals were stratified into five groups (A, B, C, D, and E) based on tumor volume and body weight on study day -2. The mean inclusion volume was 98.3 ± 8.0 mm3 (range: 39.3-191.0 mm3 and the mean (±SEM) inclusion weight was 29.6 ± 0.4 g (range: 24.8-32.7 g). No significant difference was observed for tumor volume or body weight between the groups (One-way ANOVA, p=0.9953 and p=0.9624, respectively).

[0187] Animals were dosed with either vehicle (group A) or 64Cu-DOTA-AE105 (group B, C, D, and E) at different activity levels on study day 0. Animals in group B received 111 MBq, animals in group C received 37 MBq, animals in group D received 19 MBq, and animals in group E received 37 MBq. At study day 7 and 14 animals in group E received an additional dosing with 37 MBq.

[0188] Tumor volume and body weight were monitored twice a week after the first dosing. The efficacy was evaluated over a period of 27 days. The mean tumor volumes for animals included in the study is presented in figure 4. Differences in tumor volume were evaluated at the latest timepoint where minimum 60% of animals were alive within each of the compared groups. Figure 4B show tumor volume comparison at study day 17 (the last day where 60% or more animals were alive within all groups). The effect of 19, 37 and 111 MBq Cu-64-DOTA- AE105 (Group B-D) and the effect of fractionated dosing of 37 MBq Cu-64-DOTA-AE105 (Group E) were tested against Vehicle (Group A) by One-way ANOVA with Dunnett’s multiple comparison (summary of the statistical analysis is shown in the table below, statistical significance is highlighted in bold). A significant effect was observed following treatment with 111MBq Cu-64-DOTA-AE105 compared to Vehicle and 37 MBq Cu-64-DOTA-AE105 as fractionated dosing. Bars represent mean ± SEM. N=3-5 / group.

[0189] Animals were euthanized according to humane endpoints, which was reached when tumor sizes > 1500 mm3. Kaplan-Meier curves of survival data is presented in figure 5. The survival analysis illustrated in figure 5, shows a median survival of 17 days for Vehicle and group C (37 MBq Cu-64-DOTA-AE105), a median survival of 18.5 for group D (19 MBq Cu-64-DOTA- AE105) and a median survival time of 24 days for group B and E (111 MBq and 37 (3xQW) Cu-64-DOTA-AE105).

[0190] Summary of statistical analysis of pairwise comparison of the survival is shown in the table below. All treatment groups were compared to the vehicle group by Log-rank (Mantel-Cox) test with Bonferroni correction for multiple comparison. The Bonferroni corrected threshold, used for each individual comparison, is the familywise significance level (0.05) divided by the number of comparisons performed (4). Statistical significance is highlighted in Bold.

[0191] The body weight did not differ between the groups during the study, indicating that the drug was safe and well tolerated.

[0192] As can be observed in figure 4, treatment with 64Cu-DOTA-AE105 provided a surprisingly effective reduction of tumor growth. Treatment with a single dose of 111 MBq 64Cu-DOTA- AE105 (group B) significantly and almost entirely inhibited tumor growth up to 17 days post dosing, thereby confirming the treatment efficacy observed in figure 2A even with a slightly lower dose, i.e. 111 MBq compared to 130MBq. Even more striking, it was observed that treatment with three doses of 37 MBq 64Cu-DOTA-AE105 administered once a week (group E, Figure 4) delayed tumor growth up to 17 days post first dosing at almost the same efficacy as treatment with a single dose of 111MBq 64Cu-DOTA-AE105. Thus, 64Cu-DOTA-AE105 showed significant therapeutic efficacy compared to untreated control group even at very low doses, equaling (i.e. delivering same radiation dose in Gy to the tumor) approximately 15 MBq 67Cu at which doses 67Cu labeled compound, based on available data e.g. in figure 3, would not have any therapeutic effect.

[0193] Example 6 Therapeutic effect of 64Cu-DOTA-AE105 in non-small cell lung cancer bearing xenograft mice

[0194] The treatment efficacy of 64Cu-DOTA-AE105 was tested in female NMRI nude mice bearing subcutaneous NCI-H1993 tumors.

[0195] The NCI-H1993 tumor cell line is a human non-small cell lung cancer cell line, and tumors were established subcutaneously in the flank region of the female NMRI nude mice. Ten animals were included and stratified into two groups based on tumor volume and body weight (n=5 / group).

[0196] The mean (±SEM) tumor volume at inclusion was 79.1 ± 6.9 mm3(range: 47.1-111.7 mm3) and the mean (±SEM) inclusion weight was 31 .2 ± 0.6 g (range: 27.5- 34.8 g). No significant difference was observed for tumor volume or body weight between the groups (Student's t- test p=0.98 and p=0.33, respectively).

[0197] Animals received either vehicle (untreated control) or 150 M Bq Cu-64-DOTA-AE105 at study day 0.

[0198] All mice were pre-treated with diet-gel five days prior to dosing and 5 days post dosing to prevent significant weight loss. Tumors and body weights were measured twice weekly for the duration of the study. Animals were taken down due to humane endpoint for tumor volume (> 1500mm3). The last observation for euthanized animals was carried forward until less than 60% of the animals were alive within the group.

[0199] The results are shown in figure 6, from which it can be appreciated that a single dose of 150 MBq 64Cu-DOTA-AE105 significantly delayed tumor growth for the entire duration of the study.

[0200] Tumor volumes were compared between the group treated with 64Cu-DOTA-AE105 (Group B) and Vehicle (Group C) at study day 57. The treatment with 150MBq 64Cu-DOTA-AE105 led to a significant reduction in tumor volume compared to animal receiving Vehicle (Unpaired t-test, p=0.0113).

[0201] As can be appreciated from figure 6, the 64Cu labelled uPAR binding peptide conjugate 64Cu-DOTA-AE105 stalled tumor growth for the entire duration of the study, thus providing effective treatment of human NCI-H1993 tumors.

[0202] Example 7 64Cu labelling of NODAGA-E[c(RGDyK)]2

[0203] 64Cu-NODAGA-E[c(RGDyK)]2 is produced by labelling NODAGA-E[c(RGDyK)]2 acetate with a 64Cu salt. NODAGA-E[c(RGDyK)]2 (2 nmol) was mixed with 450 pL ammonium acetate (0.1 M, pH 8.4), and 50 pL of 64CuCI2 (50-60 MBq) was added (64Cu: t1 / 2 = 12.7 h; Emax, p+ = 0.653 MeV (18%)). The solution was left at room temperature for 15 min. Example 8 Dose-response of efficacy of 64Cu-NODAGA-E[c(RGDyK)]2in glioblastoma bearing xenograft mice

[0204] The dose dependent efficacy of 64Cu-NODAGA-E[c(RGDyK)]2is tested in female NMRI nude mice bearing subcutaneous U87 MG tumors. The U87 MG tumor cell line is a human glioblastoma cell line.

[0205] Animals are stratified into five groups (A, B, C, D, and E) based on tumor volume and body weight prior to study day 0.

[0206] Animals are dosed with either vehicle (group A) or 64Cu-NODAGA-E[c(RGDyK)]2(group B, C, D, and E) at different activity levels on study day 0. Animals in group B receive 111 MBq, animals in group C receive 37 MBq, animals in group D receive 19 MBq, and animals in group E receive 37 MBq. At study day 7 and 14, animals in group E receive an additional dosing with 37 MBq.

[0207] T umor volume and body weight is monitored twice a week after the first dosing. The efficacy is evaluated over a period of up to 30 days. Differences in tumor volume is evaluated at the latest timepoint where minimum 60% of animals is alive within each of the compared groups. A therapeutic effect is observed following treatment with either 111MBq 64Cu-NODAGA- E[c(RGDyK)]2or fractionated as 3 doses of 37 MBq 64Cu-NODAGA-E[c(RGDyK)]2compared to Vehicle.

[0208] Animals are euthanized according to humane endpoints, which is reached when tumor sizes > 1500 mm3.

[0209] Example 9 Therapeutic effect of 64Cu-NODAGA-E[c(RGDyK)]2in glioblastoma bearing xenograft mice

[0210] The treatment efficacy of 64Cu-NODAGA-E[c(RGDyK)]2was tested in female NMRI nude mice bearing subcutaneous U87 MG tumors.

[0211] The U87 MG tumor cell line is a human glioblastoma cell line and tumors were established subcutaneously in the flank region of the female NMRI nude mice. Sixteen animals were included and stratified into two groups based on tumor volume and body weight (n=8 / group) on study day -1.

[0212] Animals received either vehicle (untreated control) or 111 MBq 64Cu-NODAGA- E[c(RGDyK)]2at study day 0 and study day 14.

[0213] All mice were pre-treated with diet-gel five days prior to dosing and 5 days post dosing to prevent significant weight loss. T umors and body weights were measured tree time per week for the duration of the study. Animals were taken down due to humane endpoint for tumor volume (> 1500mm3). The last observation for euthanized animals was carried forward until less than 60% of the animals were alive within the group.

[0214] The results are shown in figure 7. Figure 7A shows the mean tumor volume over 30 days in the radioligand treatment and vehicle, respectively. Figure 7B and 7C shows the data obtained from the individual mice in the radioligand therapy and vehicle, respectively. From Figure 7 A, it can be appreciated that the mean tumor growth was delayed following administration of a single dose of 111 MBq 64Cu-NODAGA-E[c(RGDyK)]2and further delayed following administration of a second dose of 111 MBq 64Cu-NODAGA-E[c(RGDyK)]2at study day 14.

[0215] Figure 7 D shows Kaplan-meier survival curves. It can be appreciated that the median survival for animals treated with 64Cu-NODAGA-E[c(RGDyK)]2was 25 days, compared to 16 days for individuals in the Vehicle group, yielding a significant increase in survival for the treated group (Log-rank (Mantel Cox) test, P= 0.0009).

Claims

CLAIMS1. A 64Cu labelled conjugate for use in treatment of cancer, wherein said 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64CuAnd wherein the targeting moiety is coupled to 64Cu by the chelating agent.

2. The 64Cu labelled conjugate for use according to claim 1 , wherein said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

3. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, CB-TE2A, or derivatives thereof.

4. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the chelating agent is selected from the group consisting of DOTA, DOTAM, NOTA, NODAGA, SARCOPHAGINE, DiAmSar, AmBaSar, MeCOSar, or CB-TE2A.

5. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the chelating agent is NODAGA.

6. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the cumulative dose is administered systemically to a human patient, preferably by the intravenous route, in the range of 40 GBq to 200 GBq, such as 40GBq to 163GBq, preferably 50GBq to 112GBq.

7. The 64Cu labelled conjugate for use according to any of claims 1 to 5, wherein the 64Cu labelled conjugate is administered locally.

8. The 64Cu labelled conjugate for use according to claim 7, wherein the 64Cu labelled conjugate is administered locally to the brain or bladder.

9. The 64Cu labelled conjugate for use according to claim 8, wherein said 64Cu labelled conjugate is administered locally to the urinary bladder by bladder installation in acumulative dose of up to 335GBq installed for 15 min to 2 h or up to 1 ,340GBq installed for 15 min or less, preferably in the range of 50-335GBq installed for 15min to 2 h or 200- 1 ,340GBq installed for 15 min or less, most preferred in the range of 100-335GBq installed for 15min to 2 h or 400-1 , 340GBq if installed for 15 min or less is administered locally to the urinary bladder.

10. The 64Cu labelled conjugate for use according to claim 9, wherein said cumulative dose is administered in single doses of 2 GBq to 10 GBq.

11. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the targeting moiety is selected from the group consisting of antibodies, antibody derived targeting moieties, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody, antibody mimetics, affibody molecules, nucleic acid molecules, aptamers, small molecules, peptides, peptidomimetics. Preferably the targeting moiety is selected from the group consisting of peptides and peptidomimetics, most preferably peptides.

12. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the targeting moiety is an RGD based targeting moiety, preferably an RGD based integrin avp3 targeting peptide.

13. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the targeting moiety is E[c(RGDyK)]2.

14. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein the 64Cu labelled conjugate has the formula:

15. The 64Cu labelled conjugate for use according to any of the preceding claims, wherein said cancer is selected from the group consisting of breast cancer, skin cancer, colorectal cancer, pancreatic cancer, prostate cancer, lung cancer, brain cancer, hepatocellular cancer, neuroendocrine cancer, ovarian cancer, renal cancer, thyroid cancer, head and neck cancer, cervical cancer, bladder cancer, astrocytomas, choriocarcinomas, glioblastomas, gliomas, hemangiomas (childhood, capillary), hepatomas, hyperplastic endometrium, Kaposi sarcoma, melanoma, neuroblastomas, seminomas.

16. A Method of treatment of a cancer disease by administering to a patient a 64Cu labelled conjugate, wherein sad 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64CuAnd wherein the targeting moiety is coupled to 64Cu by the chelating agent.

17. The method of treatment according to claim 16, wherein said 64Cu labelled conjugate is administered in a cumulative dose providing a calculated radiation dose to the cancer of less than 50 Gy, preferably less than 30Gy.

18. The method of treatment according to any of the claims 16 to 17, wherein said 64Cu labelled conjugate is administered systemically to a human patient, preferably by the intravenous route, in a cumulative dose of 40 GBq to 200 GBq, such as 40GBq to 163GBq, preferably 50GBq to 112GBq.

19. The method of treatment according to any of the claims 16 to 18, wherein the targeting moiety is selected from the group consisting of antibodies, antibody derived targeting moieties, scFv, Fab, nanobody, bispecific antibody, bifunctional antibody, diabody and minibody, antibody mimetics, affibody molecules, nucleic acid molecules, aptamers, small molecules, peptides, peptidomimetics. Preferably the targeting moiety is selected from the group consisting of peptides and peptidomimetics, most preferably peptides.

20. The method of treatment according to any of the claims 16 to 19, wherein the targeting moiety is an RGD based targeting moiety, preferably an RGD based integrin avp3 targeting peptide.

21. a pharmaceutical formulation comprising a 64Cu labelled conjugate, wherein said 64Cu labelled conjugate comprises: a) A targeting moiety b) A chelating agent suitable for binding radiometals c) The radionuclide 64CuAnd wherein the targeting moiety is coupled to 64Cu by the chelating agent.

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