Methods and reagents for targeting tumor targets with greater efficacy and less toxicity

Abstract

To provide combination therapies for reducing toxic effects of cancer treatment while maintaining or even increasing the efficacy.SOLUTION: A method involves providing a first agent comprising a first targeting component coupled to a first cancer therapeutic component, and providing a second agent comprising a second targeting component coupled to a second cancer therapeutic component. The first and second targeting components have different biodistributions and / or pharmacokinetics. The first and second agents are administered to a subject having cancer to treat the cancer. Also disclosed is a combination therapeutic agent comprising the first and second agents.SELECTED DRAWING: Figure 2

Description

Technical Field

[0001] This application claims the benefit of priority based on U.S. Provisional Patent Application No. 62 / 500,187, filed on May 2, 2017, which is hereby incorporated by reference in its entirety.

[0002] Field of the Invention The present invention relates to tumor targeting methods and reagents with greater therapeutic efficacy and less toxicity.

Background Art

[0003] Background of the Invention Combination therapy is a well - established and generally recognized treatment modality in virtually all types of cancer and has been a standard treatment for decades. The rationale for using combination therapy was the early experience in chemotherapy, where it was determined that the high mutation rate of cancer when only a single agent was used allowed for the rapid emergence of resistant strains of tumor cells. The goal of combination therapy is to increase efficacy and minimize the development of tumor resistance or escape. This is generally achieved by using two or more anticancer agents, each having a different mechanism of action, which makes it more difficult and less likely for resistant tumor cells to develop. The additive or synergistic effects obtained by combining two or more agents can make the difference between the success and failure of a patient's treatment.

[0004] In the field of oncology, many combination treatment regimens are well - known. For example, MOPP (the acronym for mechlorethamine, vincristine, procarbazine, prednisone) is a curative treatment regimen for Hodgkin's disease. Multiple different combination regimens (all including cisplatin, vinblastine, and bleomycin) have been recognized in the treatment of testicular cancer, which is curable in up to 98% of diagnosed cases. Over 300 different combination regimens have been used overall.

[0005] A major drawback of combination therapies is that they often also result in increased toxicity. For example, most forms of non-surgical cancer treatment, such as external beam radiation or chemotherapy, are limited in their effectiveness due to toxic side effects on normal tissues and cells and the limited specificity of these treatment methods for cancer cells. This limitation is also important when anti-cancer antibodies are used to target toxic agents, such as isotopes, drugs, and toxins, to the cancer site, as these circulate as systemic agents and also reach sensitive cell compartments such as the bone marrow. In acute radiation injury, destruction of the lymphatic and hematopoietic compartments is a major factor in the development of sepsis and subsequent death. Therefore, there is a growing need for methods to reduce the toxic effects of cancer treatment while maintaining or even increasing effectiveness.

[0006] The present invention relates to overcoming these and other deficiencies in the art.

Summary of the Invention

[0007] The present invention relates to a method of treating cancer. The method includes providing a first agent that includes a first targeting component bound to a first cancer treatment component and providing a second agent that includes a second targeting component bound to a second cancer treatment component. The first and second targeting components have different biodistributions and / or pharmacokinetics. The first and second agents are then administered to a subject having cancer to treat the cancer.

[0008] The present invention also relates to a combination therapy agent for treating cancer. The combination therapy agent includes a first agent that includes a first targeting component bound to a first cancer treatment component and a second agent that includes a second targeting component bound to a second cancer treatment component. The first and second targeting components have different biodistributions and / or pharmacokinetics.

[0009] The present invention has devised a method to overcome the MTD of a target agent in order to achieve improved efficacy while having the opportunity to reduce it without increasing its toxicity. The present invention proposes the use of two distinct targeted agents, each of which does not target the same molecule or cell type. In this approach, each of the two targeted agents has a different biodistribution and / or pharmacokinetics from the other. Importantly, the different biodistributions and pharmacokinetics of these respective agents result in different non-overlapping toxicities for each of the two targeted agents. When the two targeted agents are combined in a treatment strategy, as a result, both drugs accumulate at the desired target site simultaneously or sequentially, thereby bringing about a combined treatment effect. However, since the biodistributions of the two targeted agents are different and as a result drug delivery to normal tissues does not increase and toxicity does not increase, normal tissue toxicity does not increase. [The present invention 1001] Providing a first agent comprising a first targeting component bound to a first cancer treatment component; Providing a second agent comprising a second targeting component bound to a second cancer treatment component, wherein the first and second targeting components have different biodistributions and / or pharmacokinetics in a subject; said providing; Administering the first and second agents to a subject having cancer in order to treat the cancer; A method of treating cancer, comprising. [The present invention 1002] The method of the present invention 1001, wherein the first and second targeting components are independently selected from the group consisting of an antibody or a binding fragment thereof, a protein, a peptide, and a small molecule. [The present invention 1003] The method of the present invention 1001, wherein the first and second targeting components target the same molecular target. [The present invention 1004] The method of the present invention 1001, wherein the first and second targeting components target different molecular targets on the same cell. [The present invention 1005] The method of the present invention 1001, wherein the first and second cancer treatment components each have a maximum tolerated dose, and the maximum tolerated doses of the first and second cancer treatment components are administered during the administration. [The present invention 1006] The method of the present invention 1001, wherein the first and second cancer treatment components each have a maximum tolerated dose, and an amount less than the maximum tolerated doses of the first and second cancer treatment components is administered during the administration. [The present invention 1007] The method of the present invention 1001, wherein the first and second cancer treatment components are independently selected from the group consisting of radionuclides and chemotherapeutic agents. [The present invention 1008] The first and / or second cancer treatment component is 86 Re, 90 Y, 67 Cu, 169 Er, 121 Sn, 127 Te, 142 Pr, 143 Pr, 198 Au, 199 Au, 161 Tb, 109 Pd, 188 Rd, 166 Dy, 166 Ho, 149 Pm, 151 Pm, 153 Sm, 159 Gd, 172 Tm, 169 Yb, 175 Yb, 177 Lu, 105 Rh, 111 Ag, 131 I, 177m Sn, 225 Ac, 227 Th, 211 The method of the present invention 1007, which is a radionuclide independently selected from the group consisting of At, and combinations thereof. [The present invention 1009] The method of the present invention 1007, wherein the first and / or second cancer treatment components are chemotherapeutic agents independently selected from the group consisting of busulfan, cisplatin, carboplatin, chlorambucil, cyclophosphamide, ifosfamide, dacarbazine (DTIC), mechlorethamine (nitrogen mustard), melphalan, carmustine (BCNU), lomustine (CCNU), 5-fluorouracil (5-FU), capecitabine, methotrexate, gemcitabine, cytarabine (ara-C), fludarabine, dactinomycin, daunorubicin, doxorubicin (adriamycin), idarubicin, mitoxantrone, paclitaxel, docetaxel, etoposide (VP-16), vinblastine, vincristine, vinorelbine, prednisone, dexamethasone, tamoxifen, fulvestrant, anastrozole, letrozole, megestrol acetate, bicalutamide, flutamide, leuprolide, goserelin, L-asparaginase, tretinoin, maytansine, auristatin, pyrrolobenzodiazepine, duocarmycin, and combinations thereof. [The present invention 1010] The method of the present invention 1001, wherein the cancer is prostate cancer. [The present invention 1011] The method of the present invention 1010, wherein the first and second target-directed components target the prostate-specific membrane antigen (PSMA) receptor. [The present invention 1012] The method of the present invention 1011, wherein the first target-directed component is a PSMA receptor antibody or derivative against the PSMA receptor, and the second target-directed component is a PSMA receptor-binding peptide or a PSMA receptor inhibitor. [The present invention 1013] The method of the present invention 1012, wherein the first target-directed component is an antibody selected from the group consisting of J591, J415, J533, and E99, while the second target-directed component is a peptide selected from the group consisting of PSMA 617, PSMA I&T, DCFBC, DCFPyL, glutamate-urea-lysine analog, phosphoramidate analog, 2-(phosphinylmethyl)pentanedioic acid analog, and other PSMA ligands / inhibitors. [The present invention 1014] The first agent is J591- 177 Lu, and the second agent is PSMA 617- 177 Lu or PSMA I&T- 177 Lu, the method of the present invention 1012. [The present invention 1015] The method of the present invention 1001, wherein the subject is human. [The present invention 1016] The method of the present invention 1001, wherein the cancer is a neuroendocrine cancer. [The present invention 1017] The method of the present invention 1016, wherein the first and second target-directed components target somatostatin receptors. [The present invention 1018] The method of the present invention 1017, wherein the first and second target-directed components target somatostatin receptor-2 isoform. [The present invention 1019] The method of the present invention 1016, wherein the neuroendocrine cancer is selected from the group consisting of carcinoid tumor, gastrinoma, insulinoma, glucagonoma, VIPoma, somatostatinoma, thyroid cancer, Merkel cell cancer of the skin, tumor of the anterior pituitary gland, medullary cancer, parathyroid tumor, thymic and mediastinal carcinoid tumor, pulmonary neuroendocrine tumor, adrenal medullary tumor, pheochromocytoma, schwannoma, paraganglioma, neuroblastoma, and urothelial carcinoid neuroendocrine cancer. [The present invention 1020] The method of the present invention 1001, wherein the cancer is breast cancer. [The present invention 1021] The method of the present invention 1020, wherein the first and second target-directed components target the HER receptor family. [The present invention 1022] The method of the present invention 1001, wherein the cancer is non-Hodgkin lymphoma. [The present invention 1023] The method of the present invention 1022, wherein the first and second target-directed components target CD20. [The present invention 1024] The method of the present invention 1001, wherein the first agent and the second agent are different. [The present invention 1025] The method of the present invention 1001, wherein the first and second target-directed components target a cancer cell receptor. [The present invention 1026] A first agent comprising a first target-directed component bound to a first cancer treatment component, A second agent comprising a second target-directed component bound to a second cancer treatment component A combination therapy agent for treating cancer, comprising: wherein the first and second targeting components have different biodistributions and / or pharmacokinetics, Combination therapy agent. [The present invention 1027] The combination therapy agent of the present invention 1026, wherein the first and second target-directed components are independently selected from the group consisting of an antibody or a binding fragment thereof, a protein, a peptide, and a small molecule. [The present invention 1028] The combination therapy agent of the present invention 1026, wherein the first and second target-directed components target the same molecular target. [The present invention 1029] The combination therapy agent of the present invention 1026, wherein the first and second target-directed components target different molecular targets on the same cell. [The present invention 1030] The combination therapy agent of the present invention 1026, wherein the first and second cancer treatment components are independently selected from the group consisting of a radionuclide and a chemotherapeutic agent. [The present invention 1031] The first and / or second cancer treatment component is 86 Re, 90 Y, 67 Cu, 169 Er,121 Sn, 127 Te, 142 Pr, 143 Pr, 198 Au, 199 Au, 161 Tb, 109 Pd, 188 Rd, 166 Dy, 166 Ho, 149 Pm, 151 Pm, 153 Sm, 159 Gd, 172 Tm, 169 Yb, 175 Yb, 177 Lu, 105 Rh, 111 Ag, 131 I, 177m Sn, 225 Ac, 227 Th, 211 The combined therapeutic agent of the present invention 1030, which is a radionuclide independently selected from the group consisting of At and combinations thereof. [The present invention 1032] The first and / or second cancer treatment components are busulfan, cisplatin, carboplatin, chlorambucil, cyclophosphamide, ifosfamide, dacarbazine (DTIC), mechlorethamine (nitrogen mustard), melphalan, carmustine (BCNU), lomustine (CCNU), 5-fluorouracil (5-FU), capecitabine, methotrexate, gemcitabine, cytarabine (ara-C), fludarabine, dactinomycin, daunorubicin, doxorubicin (adriamycin), idarubicin, mitoxantrone, paclitaxel, docetaxel, etoposide (VP-16), vinblastine, vincristine, vinorelbine, prednisone, dexamethasone, tamoxifen, fulvestrant, anastrozole, letrozole, megestrol acetate, bicalutamide, flutamide, leuprolide, goserelin, L-asparaginase, tretinoin, maytansine, auristatin, pyrrolobenzodiazepine, duocarmycin, and combinations thereof. The combined therapeutic agent of the present invention 1030, which is a chemotherapeutic agent independently selected from the group consisting of these. [The present invention 1033] The combination therapy agent of the present invention 1026, wherein the cancer is prostate cancer. [The present invention 1034] The combination therapy agent of the present invention 1033, wherein the first and second target - directed components target the prostate - specific membrane antigen (PSMA) receptor. [The present invention 1035] The combination therapy agent of the present invention 1034, wherein the first target - directed component is a PSMA receptor antibody or derivative against the PSMA receptor, and the second target - directed component is a PSMA receptor - binding peptide or PSMA receptor inhibitor. [The present invention 1036] The combination therapy agent of the present invention 1035, wherein the first target - directed component is an antibody selected from the group consisting of J591, J415, J533, and E99, while the second target - directed component is a peptide selected from the group consisting of PSMA 617, PSMA I&T, DCFBC, DCFPyL, glutamate - urea - lysine analog, phosphoramidate analog, 2 - (phosphinylmethyl)pentanedioic acid analog, and other PSMA ligands / inhibitors. [The present invention 1037] The first agent is J591 - 177 Lu, and the second agent is PSMA 617 - 177 Lu or PSMA I&T - 177 Lu, which is the combination therapy agent of the present invention 1035. [The present invention 1038] The combination therapy agent of the present invention 1026, wherein the cancer is neuroendocrine cancer. [The present invention 1039] The combination therapy agent of the present invention 1038, wherein the first and second agents target the somatostatin receptor. [The present invention 1040] The combination therapy agent of the present invention 1039, wherein the first and second target - directed components target the somatostatin receptor - 2 isoform. [The present invention 1041] The combined therapeutic agent of the present invention 1038, wherein the neuroendocrine cancer is selected from the group consisting of carcinoid tumor, gastrinoma, insulinoma, glucagonoma, VIPoma, somatostatinoma, thyroid cancer, Merkel cell cancer of the skin, tumor of the anterior pituitary gland, medullary cancer, parathyroid tumor, thymic and mediastinal carcinoid tumor, pulmonary neuroendocrine tumor, adrenal medullary tumor, pheochromocytoma, schwannoma, paraganglioma, neuroblastoma, and urinary tract carcinoid neuroendocrine cancer. [The present invention 1042] The combined therapeutic agent of the present invention 1026, wherein the cancer is breast cancer. [The present invention 1043] The combined therapeutic agent of the present invention 1042, wherein the first and second target-directed components target the HER receptor family. [The present invention 1044] The combined therapeutic agent of the present invention 1026, wherein the cancer is non-Hodgkin lymphoma. [The present invention 1045] The combined therapeutic agent of the present invention 1044, wherein the first and second target-directed components target CD20. [The present invention 1046] The combined therapeutic agent of the present invention 1026, wherein the first drug and the second drug are different. [The present invention 1047] The combined therapeutic agent of the present invention 1026, wherein the first and second target-directed components target cancer cell receptors.

Brief Description of the Drawings

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[0011] The present invention relates to a method for treating cancer. The method includes providing a first agent that includes a first targeting component bound to a first cancer treatment component and providing a second agent that includes a second targeting component bound to a second cancer treatment component. The first and second targeting components have different biodistributions and / or pharmacokinetics. The first and second agents are then administered to a subject having cancer to treat the cancer.

[0012] As used herein, the term "treating" refers to applying or administering the first and second agents of the present invention to a subject, such as a patient. Treating can be curing, treating, reducing, alleviating, changing, modifying, inducing remission, soothing, improving cancer, cancer symptoms, or a tendency to be susceptible to cancer, or acting on cancer, cancer symptoms, or a tendency to be susceptible to cancer.

[0013] As used herein, the term "subject" is intended to include humans and non-human animals. Non-human animals include all vertebrates such as, for example, mammals and non-mammals (non-human primates, sheep, dogs, cows, chickens, amphibians, reptiles, etc.).

[0014] As used herein, the term "cancer" includes all types of cancerous growths or carcinogenic processes, metastatic tissues, or malignantly transformed cells, tissues, or organs, regardless of histopathological type or stage of invasiveness.

[0015] As used herein, the term "maximum tolerated dose (MTD)" refers to the dose of a therapeutic drug that includes a target drug in an amount such that unacceptable toxicity occurs if exceeded. This applies regardless of whether the drug targets a specific cell type or a specific molecule. Due to the MTD and the limits of drug (target drug or otherwise) tolerance, the maximum anti-cancer effect cannot usually be achieved. The MTD of a drug is greatly affected by its biodistribution and pharmacokinetics.

[0016] As used herein, the term "biodistribution" refers to the organs and tissues in which a drug is distributed in the body.

[0017] As used herein, the term "pharmacokinetics" refers to the length of time a drug remains in the body.

[0018] In certain embodiments, the cancer is prostate cancer, neuroendocrine cancer, breast cancer, or non-Hodgkin lymphoma. In some embodiments, the cancer is a primary tumor, and in another embodiment, the cancer is a secondary or metastatic tumor.

[0019] As used herein, a "targeting moiety" is a moiety that can bind to or otherwise associate with a molecular target such as, for example, a membrane component, a cell surface receptor, prostate specific membrane antigen (also known as PSMA, folate hydrolase 1, glutamate carboxypeptidase II, and NAALADase). The first and second agents comprising the targeting moiety may localize or concentrate at a particular target site such as, for example, a tumor, a diseased site, a tissue, an organ, certain cells, etc. Thus, the first and second agents may be "target specific". In some instances, a therapeutic agent may exert its anti-cancer effect without the need for release from the targeting moiety. In another instance, a therapeutic component may be released from the first and / or second agent and may be able to interact locally with a particular target site.

[0020] For example, exemplary targeting moieties can include nucleic acids, peptides, polypeptides, proteins, glycoproteins, carbohydrates, or lipids. The targeting moiety may be a natural or synthetic ligand of a cell surface receptor such as, for example, a growth factor, a hormone, LDL, transferrin, etc. The targeting moiety may be an antibody, and this term is intended to include antibody fragments, characteristic portions of an antibody, such as single-chain targeting portions that can be identified using procedures such as phage display. The targeting moiety may be a naturally occurring or artificially formed (e.g., by chemical synthesis) targeting peptide, targeting peptide mimetic, or small molecule.

[0021] In one embodiment, the first and second targeting moieties are independently selected from the group consisting of an antibody or a binding fragment thereof, a protein, a peptide, an oligonucleotide, and a small molecule.

[0022] Antibodies against molecular targets on tumors are known. For example, antibodies and antibody fragments that specifically bind to markers produced by or associated with tumors are disclosed in, inter alia, Hansen U.S. Patent No. 3,927,193, and Goldenberg U.S. Patents Nos. 4,331,647, 4,348,376, 4,361,544, 4,468,457, 4,444,744, 4,818,709, and 4,624,846, the entire contents of all of which are incorporated herein by reference. In particular, antibodies against antigens such as, for example, gastrointestinal, lung, breast, prostate, ovarian, testicular, brain, or lymphatic tumors, sarcomas, or melanomas are advantageously used. Antibodies against cancer-related antigens are well known to those skilled in the art.

[0023] The antibodies of the present invention may exist in various forms such as, for example, polyclonal antibodies, monoclonal antibodies, intracellular antibodies (“intrabodies”), antibody fragments (e.g., Fv, Fab, and F(ab)2), half-antibodies, hybrid derivatives, as well as single-chain antibodies (scFv), chimeric antibodies, and humanized antibodies (Ed Harlow and David Lane, USING ANTIBODIES: A LABORATORY MANUAL (Cold Spring Harbor Laboratory Press, 1999); Houston et al., “Protein Engineering of Antibody Binding Sites: Recovery of Specific Activity in an Anti-Digoxin Single-Chain Fv Analogue Produced in Escherichia coli,” Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); Bird et al, “Single-Chain Antigen-Binding Proteins,” Science 242:423-426 (1988), each of which is incorporated herein by reference in its entirety).

[0024] The antibody of the present invention may be a synthetic antibody. A synthetic antibody is an antibody produced using recombinant DNA technology, such as an antibody expressed by a bacteriophage, for example. Alternatively, a synthetic antibody is produced by synthesis of a DNA molecule encoding and expressing the antibody of the present invention, or synthesis of an amino acid sequence defining the antibody, where the DNA or amino acid sequence in this case is obtained using synthetic DNA or amino acid sequence techniques well known in the art.

[0025] The method for monoclonal antibody production can be carried out using the techniques described herein, or this is well known in the art (MONOCLONAL ANTIBODIES - PRODUCTION, ENGINEERING AND CLINICAL APPLICATIONS (Mary A. Ritter and Heather M. Ladyman eds., 1995), which is hereby incorporated by reference in its entirety). Usually, the method involves obtaining immune cells (lymphocytes) from the spleen of a mammal pre-immunized with an antigen of interest either in vivo or in vitro.

[0026] Alternatively, monoclonal antibodies can be produced using recombinant DNA methods such as those described in U.S. Patent No. 4,816,567 to Cabilly et al., which is hereby incorporated by reference in its entirety. Polynucleotides encoding monoclonal antibodies are isolated from mature B cells or hybridoma cells by RT-PCR using, for example, oligonucleotide primers that specifically amplify the genes encoding the heavy and light chains of the antibody. The isolated polynucleotides encoding the heavy and light chains are then cloned into an appropriate expression vector and transfected into host cells such as E. coli cells, monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin proteins, whereupon the monoclonal antibody is produced by the host cells. Also, recombinant monoclonal antibodies or fragments thereof of the desired species can be isolated from phage display libraries (McCafferty et al., “Phage Antibodies: Filamentous Phage Displaying Antibody Variable Domains,” Nature 348:552-554 (1990); Clackson et al., “Making Antibody Fragments using Phage Display Libraries,” Nature 352:624-628 (1991); and Marks et al., “By-Passing Immunization. Human Antibodies from V-Gene Libraries Displayed on Phage,” J. Mol. Biol. 222:581-597 (1991), which are hereby incorporated by reference in their entirety).

[0027] Polynucleotides encoding monoclonal antibodies can be further modified using recombinant DNA techniques to generate alternative antibodies. For example, the constant domains of the light and heavy chains of a murine monoclonal antibody can be replaced with those of a human antibody to generate a chimeric antibody. Alternatively, the constant domains of the light and heavy chains of a murine monoclonal antibody can be replaced with non-immunoglobulin polypeptides to generate a fusion antibody. In another embodiment, the constant region is truncated or removed to generate the desired antibody fragment of the monoclonal antibody. Furthermore, site-specific or high-density mutagenesis of the variable region can be used to optimize the specificity and affinity of the monoclonal antibody.

[0028] The monoclonal antibodies of the present invention may be humanized antibodies. A humanized antibody is an antibody that contains a minimal sequence derived from a non-human (e.g., murine) antibody within the variable region. Such antibodies are used therapeutically to reduce antigenicity and the human anti-mouse antibody response when administered to a human subject. In practice, a humanized antibody is typically a human antibody with minimal or no non-human sequences. A human antibody is an antibody produced by a human or an antibody having an amino acid sequence corresponding to an antibody produced by a human.

[0029] In addition to the whole antibody, the present invention also encompasses the binding portion of such an antibody. Such binding portions include monovalent Fab fragments, Fv fragments (e.g., single-chain antibodies, scFv), and single variable V H and V LDomains, as well as divalent F(ab’)2 fragments, Bis-scFv, diabodies, triabodies, minibodies, and the like are included. These antibody fragments can be produced by conventional techniques such as proteolytic fragment formation methods as described in James Goding, MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE 98 - 118 (Academic Press, 1983) and Ed Harlow and David Lane, ANTIBODIES: A LABORATORY MANUAL (Cold Spring Harbor Laboratory, 1988) (which are hereby incorporated by reference in their entirety), or by other techniques known in the art.

[0030] Furthermore, particularly in the case of antibody fragments, it may be desirable to modify the antibody to increase its serum half-life. This can be achieved, for example, by incorporating a salvage receptor binding epitope into the antibody fragment by mutation of an appropriate region in the antibody fragment, or by fusion to the antibody fragment (e.g., by DNA or peptide synthesis) either at the terminus or in the middle after incorporation of the epitope into a peptide tag.

[0031] Antibody mimetics are also suitable for use according to the present invention. Without limitation, the 10th human fibronectin type III domain ( 10Monobodies derived from Fn3) that are known in the art (Koide et al., “The Fibronectin Type III Domain as a Scaffold for Novel Binding Proteins,” J. Mol. Biol. 284:1141-1151 (1998); Koide et al., “Probing Protein Conformational Changes in Living Cells by Using Designer Binding Proteins: Application to the Estrogen Receptor,” Proc. Natl. Acad. Sci. USA 99:1253-1258 (2002), each of which is incorporated herein by reference in its entirety); and affibodies derived from the stable α-helix bacterial receptor domain Z of staphylococcal protein A that are known in the art (Nord et al., “Binding Proteins Selected from Combinatorial Libraries of an alpha-helical Bacterial Receptor Domain,” Nature Biotechnol. 15(8):772-777 (1997), which is incorporated herein by reference in its entirety), and many other antibody mimetics are known in the art.

[0032] The peptides used in connection with the present invention can be obtained by known isolation and purification protocols from natural sources, synthesized by standard solid-phase or liquid-phase peptide synthesis methods according to known peptide sequences of the peptides, or obtained from commercially available formulations. This specification includes peptides that exhibit the biological binding properties of natural peptides and retain the specific binding properties of natural peptides. Derivatives and analogs of the peptides used herein include modifications to the composition, identity, and derivatization of the individual amino acids of the peptide, provided that the peptide retains the specific binding properties of the natural peptide. Examples of such modifications include any amino acid modification that includes D-stereoisomers, substitution of the aromatic side chains of aromatic amino acids, derivatization of the amino or carboxyl groups of the side chains of amino acids that contain such groups in their side chains, substitution of the amino or carboxyl terminus of the peptide, attachment of the peptide to a second peptide or biologically active moiety, and cyclization of the peptide (G. Van Binst and D. Tourwe, “Backbone Modifications in Somatostatin Analogues: Relation Between Conformation and Activity,” Peptide Research 5:8-13 (1992), which is hereby incorporated by reference in its entirety).

[0033] In one embodiment, the first and second target-directed components target the same molecular target. For example, the first and second target-directed components can bind to the same receptor (e.g., PSMA) expressed by the same cell type.

[0034] In another embodiment, the first and second target-directed components target different molecular targets on the same cell type. For example, the first and second target-directed components can bind to different receptors (e.g., HER1 and HER2) expressed in the same cell type.

[0035] As used herein, the "cancer treatment component" is an agent or combination of agents that treats a cell, tissue, or subject having a condition requiring treatment when contacted with the cell, tissue, or subject. The first and second cancer treatment components may be the same or different, and may be, for example, a therapeutic radionuclide, a chemotherapeutic agent, a hormone, a hormone antagonist, a receptor antagonist, an enzyme or proenzyme activated by another agent, a biologic, an autocrine, or a cytokine. Toxins can also be used in the methods of the present invention. Other therapeutic agents useful in the present invention include radiolabeled oligonucleotides such as anti-DNA, anti-RNA, antisense oligodeoxyribonucleotides, anti-protein and anti-chromatin cytotoxic agents or antibacterial agents. Other therapeutic agents are known to those skilled in the art, and the use of such other therapeutic agents according to the present invention is expressly contemplated.

[0036] The first and second cancer treatment components may be the same, but in one embodiment they are different. For example, the first and second cancer treatment components may contain different radionuclides, or the second cancer treatment component may contain a radionuclide while the first cancer treatment component contains a chemotherapeutic agent, or the second cancer treatment component may contain a chemotherapeutic agent while the first cancer treatment component contains a radionuclide.

[0037] In one embodiment, the first and second cancer treatment components are independently selected from the group consisting of radionuclides and chemotherapeutic agents.

[0038] In one embodiment, the first and / or second cancer treatment component is 86 Re, 90 Y, 67 Cu, 169 Er, 121 Sn, 127 Te, 142 Pr, 143 Pr, 198 Au, 199 Au, 161 Tb, 109 Pd, 188 Rd, 166 Dy, 166Ho, 149 Pm, 151 Pm, 153 Sm, 159 Gd, 172 Tm, 169 Yb, 175 Yb, 177 Lu, 105 Rh, 111 Ag, 131 I, 177 mSn, 225 Ac, 227 Th, 211 At, and radionuclides independently selected from the group consisting of these combinations.

[0039] Procedures for labeling drugs with radioisotopes are generally known in the art. For example, there are a wide variety of moieties that can function as chelating ligands and can be derivatized to the targeting component of the present invention. For example, chelating ligands may be derivatives of 1,4,7,10-tetraazacyclododecane tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), diethylenetriaminepentaacetic acid (DTPA), and 1-p-isothiocyanato-benzyl-methyl-diethylenetriaminepentaacetic acid (ITC-MX). These chelating agents typically have a group on the side chain, whereby the chelating agent can be used for linking to the targeting component of the present invention. Such a group includes, for example, benzyl isothiocyanate, whereby DOTA, DTPA, or EDTA can be attached to, for example, the amine group of the targeting component. Procedures for iodinating biological agents such as antibodies, their binding portions, probes, or ligands are described in Hunter and Greenwood, “Preparation of Iodine-131 Labelled Human Growth Hormone of High Specific Activity,” Nature 144:496-496 (1962), David et al., “Protein Iodination With Solid State Lactoperoxidase,” Biochemistry 13:1014-1021 (1974), as well as U.S. Patent No. 3,867,517 to Ling and U.S. Patent No. 4,376,110 to David, which are hereby incorporated by reference in their entirety.Another procedure for iodinating biological agents is described in Greenwood et al., “The Preparation of I-131-Labelled Human Growth Hormone of High Specific Radioactivity,” Biochem. J. 89:114-123 (1963); Marchalonis, “An Enzymic Method for the Trace Iodination of Immunoglobulins and Other Proteins,” Biochem. J. 113:299-305(1969); and Morrison et al., “Use of Lactoperoxidase Catalyzed Iodination in Immunochemical Studies,” Immunochemistry 8:289-297(1971), which are hereby incorporated by reference in their entirety. 99m The procedure for Tc labeling is described in Rhodes, B. et al. in Burchiel, S. et al. (eds.), Tumor Imaging: The Radioimmunochemical Detection of Cancer, New York: Masson 111-123(1982) and the references cited therein, which are hereby incorporated by reference in their entirety. 111Procedures suitable for labeling biological agents are described in Hnatowich et al., “The Preparation of DTPA-coupled Antibodies Radiolabeled With Metallic Radionuclides: an Improved Method,” J. Immul. Methods 65:147-157 (1983), Hnatowich et al., “Coupling Antibody With DTPA--an Alternative to the Cyclic Anhydride,” Int. J. Applied Radiation 35:554-557 (1984), and Buckley et al., “An Efficient Method For Labelling Antibodies With 111In,” F.E.B.S. 166:202-204 (1984), which are hereby incorporated by reference in their entirety.

[0040] In another embodiment, the first and / or second cancer treatment component is an anti-cancer agent independently selected from the group consisting of busulfan, cisplatin, carboplatin, chlorambucil, cyclophosphamide, ifosfamide, dacarbazine (DTIC), mechlorethamine (nitrogen mustard), melphalan, carmustine (BCNU), lomustine (CCNU), 5-fluorouracil (5-FU), capecitabine, methotrexate, gemcitabine, cytarabine (ara-C), fludarabine, dactinomycin, daunorubicin, doxorubicin (adriamycin), idarubicin, mitoxantrone, paclitaxel, docetaxel, etoposide (VP-16), vinblastine, vincristine, vinorelbine, prednisone, dexamethasone, tamoxifen, fulvestrant, anastrozole, letrozole, megestrol acetate, bicalutamide, flutamide, leuprolide, goserelin, L-asparaginase, tretinoin, maytansine, auristatin, pyrrolobenzodiazepine, duocarmycin, and combinations thereof.

[0041] Procedures for conjugating biological agents with chemotherapeutic agents are well known in the art. Most of the chemotherapeutic agents currently used in the treatment of cancer have functional groups that are readily capable of direct chemical cross-linking with the amine or carboxyl groups of the targeting components of the present invention. For example, in methotrexate, doxorubicin, daunorubicin, cytarabine, cisplatin, vindesine, mitomycin, and bleomycin, free amino groups are available, while in methotrexate, melphalan, and chlorambucil, free carboxylic acid groups are available. These functional groups, which are free amino and carboxylic acids, are the targets of various homo- and hetero-bifunctional chemical cross-linking agents capable of directly cross-linking these drugs to the free amino groups of the targeting components. Specific procedures for conjugating the targeting components with chemotherapeutic agents are described and are known in the art. For example, the conjugation of chlorambucil with an antibody is described in Flechner, “The Cure and Concomitant Immunization of Mice Bearing Ehrlich Ascites Tumors by Treatment With an Antibody--Alkylating Agent Complex,” European Journal of Cancer 9:741-745 (1973); Ghose et al., “Immunochemotherapy of Cancer with Chlorambucil-Carrying Antibody,” British Medical Journal 3:495-499(1972); and Szekerke et al., “The Use of Macromolecules as Carriers of Cytotoxic Groups (part II)Nitrogen Mustard--Protein Complexes,” Neoplasma 19:211-215(1972), which are hereby incorporated by reference in their entirety.The procedure for conjugating daunomycin and adriamycin to an antibody is described in Hurwitz et al., “The Covalent Binding of Daunomycin and Adriamycin to Antibodies, With Retention of Both Drug and Antibody Activities,” Cancer Research 35:1175-1181 (1975), and Arnon et al. Cancer Surveys 1:429-449 (1982), which are hereby incorporated by reference in their entirety. The coupling procedure is also described in EP86309516.2, which is hereby incorporated by reference in its entirety.

[0042] It will be understood that the exact dosages of the first and second agents of the present invention will be selected by the individual physician taking into account the patient to be treated. Usually, the dosage and administration are adjusted to provide an effective amount of the agent to the patient being treated. As used herein, an “effective amount” of an agent refers to the amount necessary to produce the desired biological response. As will be understood by those skilled in the art, the effective amount of the agents of the present invention can vary depending on factors such as the desired biological endpoint, the drug being delivered, the target tissue, the route of administration, etc. For example, an effective amount of an agent containing an anti-cancer agent will be the amount that results in reducing the size of the tumor by the desired amount over the desired period. Additional factors that may be considered include the severity of the condition; the age, weight, and gender of the patient being treated; diet therapy, the time and frequency of administration; drug combinations; response sensitivity; and tolerance / response to the treatment.

[0043] Typically, when administered as a single agent, the dosage can range from about 25% to about 100% of the MTD of the target agent. Based on the composition, the dosage can be delivered once, continuously by means such as a continuous pump, or at regular intervals. The dosage can be appropriately adjusted to achieve the desired drug level locally or systemically. If the response in the subject at such a dosage is insufficient, higher dosages (or higher effective dosages by different, more local delivery routes) can be used within the range tolerated by the patient. To achieve an appropriate systemic level of the compound, continuous IV administration over, for example, 24 hours or multiple administrations per day are also envisioned.

[0044] In one embodiment, the first and second cancer treatment components each have a maximum tolerated dose, and the maximum tolerated doses of the first and second cancer treatment components are administered to the subject. Since the biodistribution and pharmacokinetics are different for the two targeted components, the toxicities of these as individual drugs do not overlap or only minimally overlap. As a result, the increased additive dosage to the target site is not accompanied by a commensurate increase in toxicity.

[0045] In another embodiment, less than the maximum tolerated dose of the first and second cancer treatment components is administered to the subject. When the two treatment components are combined in amounts less than the maximum tolerated dose in a treatment strategy, both drugs accumulate at the target site of interest (simultaneously or sequentially), resulting in an additive treatment effect, but the subject experiences lower toxicity since the agents are administered below their MTD.

[0046] In one embodiment, the first agent is an antibody conjugated to a radionuclide and is administered at a dose of about 100 - 160 mCi total over a 2 - week cycle, such as a dose of 100, 110, 120, 130, 140, 150, or 160 mCi total over a 2 - week cycle.

[0047] In another embodiment, the first agent is an antibody conjugated to a radionuclide and is administered at a dose of about 120 - 140 mCi total over a 2 - week cycle, such as a dose of 120, 125, 130, or 140 mCi total over a 2 - week cycle.

[0048] In a further embodiment, the second agent is a small molecule conjugated to a radionuclide and is administered in a two-week cycle at a total dose of about 300 - 500 mCi in a two-week cycle, such as a dose of 300, 325, 350, 375, 400, 425, 450, 475, or 500 mCi.

[0049] In practicing the methods of the present invention, the administration step is carried out to treat the subject's cancer. In one embodiment, a subject having cancer is selected prior to the administration step. Such administration can be carried out systemically or by direct or local administration to the tumor site. For example, suitable modes of systemic administration include, but are not limited to, oral, topical, transdermal, parenteral, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, or intranasal injection, intravesical or intracavitary instillation, intravitreal, intraarterial, intralesional, or mucosal application. Suitable modes of local administration include, but are not limited to, catheter insertion, implantation, direct injection, skin / transdermal application, or portal vein administration to the relevant tissue, or any other local administration technique, method, or procedure generally known in the art. The mode of delivery that affects the therapeutic agent will vary depending on the type of therapeutic agent (e.g., antibody or inhibitory nucleic acid molecule) and the disease being treated.

[0050] The medicament of the present invention can be administered orally, for example, together with an inert diluent or with an absorbable edible carrier, or it may be enclosed in hard or soft shell capsules, or it may be compressed into tablets, or they may be directly incorporated into the foods of diet therapy. The medicament of the present invention can also be administered in a sustained release manner incorporated in devices such as sustained release capsules and nanotubes. Such devices provide flexibility with respect to time and dosage. For oral therapeutic administration, the medicament of the present invention is incorporated with excipients and can be used in the form of tablets, capsules, elixirs, suspensions, syrups and the like. Such compositions and formulations should contain at least 0.1% of the medicament, but lower concentrations can be effective and may actually be optimal. The proportion of the medicament in these compositions can of course vary and can conveniently be about 2% to about 60% by unit weight. The amount of the medicament of the present invention in such therapeutically useful compositions is an amount such that an appropriate dosage is obtained.

[0051] When the medicament of the present invention is administered parenterally, a solution or suspension of the medicament can be prepared in water appropriately mixed with a surfactant such as hydroxypropylcellulose. The dispersion can also be prepared in glycerol in oil, liquid polyethylene glycol, and mixtures thereof. Exemplary oils are of petroleum, animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, or mineral oil. Usually, water, physiological saline, aqueous dextrose solutions and related sugar solutions, and glycols such as propylene glycol and polyethylene glycol are preferred liquid carriers, especially for injection solutions. Under normal storage and use conditions, these formulations contain preservatives to prevent the growth of microorganisms.

[0052] Suitable pharmaceutical preparations for injection use include sterile aqueous solutions or dispersions, and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid to the extent that it can be easily injected. It must be stable under the conditions of manufacture and storage, and must be protected from the contaminating action of microorganisms such as bacteria and fungi. The carrier may be a solvent or dispersion medium including, for example, water, ethanol, polyols (such as glycerol, propylene glycol, and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.

[0053] When it is desired to deliver the agents of the present invention systemically, they may be formulated for parenteral administration by injection, such as by bolus injection or continuous infusion. Injectable formulations may be provided in unit dosage forms, such as in ampoules or multi-dose containers, with added preservatives. The composition may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may also contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents.

[0054] Intraperitoneal or intrathecal administration of the agents of the present invention can also be achieved using an infusion pump device. Such devices allow for continuous infusion of the target compound, avoiding multiple injections and multiple manipulations.

[0055] In addition to the formulations described above, the agents may be formulated as depot formulations. Such long-acting formulations may be formulated using suitable polymeric or hydrophobic materials (such as emulsions in acceptable oils) or ion exchange resins, or as poorly soluble derivatives, such as, for example, as poorly soluble salts.

[0056] According to one embodiment of the present invention, the cancer is prostate cancer.

[0057] In another embodiment of this aspect of the present invention, when the cancer is prostate cancer, the first and second target-directed components target the PSMA receptor.

[0058] As used herein, the "PSMA" or "prostate-specific membrane antigen" protein refers to mammalian PSMA, preferably human PSMA protein. The long transcript of PSMA encodes a protein product with a molecular weight of about 100-120 kDa, which is characterized as a type II transmembrane receptor having sequence homology with the transferrin receptor and having NAALADase activity (Carter et al., "Prostate-Specific Membrane Antigen is a Hydrolase With Substrate and Pharmacologic Characteristics of a Neuropeptidase," Proc. Natl. Acad. Sci. USA 93:749-753 (1996), which is incorporated herein by reference in its entirety).

[0059] In another embodiment, the first targeting component is a PSMA receptor antibody and the second targeting component is a PSMA receptor binding peptide or a PSMA receptor inhibitor.

[0060] A PSMA receptor antibody is an antibody that interacts (e.g., binds) with PSMA, preferably human PSMA protein. Preferably, the PSMA receptor antibody interacts (e.g., binds) with the extracellular domain of PSMA, such as the extracellular domain of human PSMA located at amino acids about 44-750 of human PSMA (the amino acid residues corresponding to the human PSMA sequence disclosed in U.S. Patent No. 5,538,866, which is incorporated herein by reference in its entirety). PSMA receptor antibodies are known in the art (Goldsmith et al., "Targeted Radionuclide Therapy for Prostate Cancer," in Therapeutic Nuclear Medicine 617-628 (R. Baum ed. 2014), which is incorporated herein by reference in its entirety). Exemplary PSMA receptor antibodies include, but are not limited to, J591, J415, J533, and E99.

[0061] The PSMA receptor inhibitor may include lipids, carbohydrates, polynucleotides, peptides, polypeptides, or any other biological, organic, or inorganic molecule that inhibits the function of the PSMA receptor. Exemplary PSMA receptor inhibitors are known in the art and include, but are not limited to, PSMA 617, PSMA I&T, DCFBC, DCFPyL, glutamate-urea-lysine analogs, phosphoramidate analogs, and 2-(phosphinylmethyl)pentanedioic acid analogs (Lutje et al., “PSMA Ligands for Radionuclide Imaging and Therapy of Prostate Cancer: Clinical Status,” Theranostics 5(12):1388-1401(2015); Haberkorn et al., “New Strategies in Prostate Cancer: Prostate-Specific Membrane Antigen (PSMA) Ligands for Diagnosis and Therapy,” Clin. Cancer Res. 22(1):9-15(2016), which are incorporated herein by reference in their entirety).

[0062] In one embodiment, the PSMA receptor antibody is selected from the group consisting of J591, J415, J533, and E99, while the second targeting component is a peptide selected from the group consisting of PSMA 617, PSMA I&T, DCFBC, DCFPyL, glutamate-urea-lysine analogs, phosphoramidate analogs, 2-(phosphinylmethyl)pentanedioic acid analogs, and other PSMA ligands / inhibitors.

[0063] In one embodiment, the first agent is J591- 177 Lu, and the second agent is PSMA 617- 177 Lu or PSMA I&T- 177It is Lu. For example, a PSMA receptor antibody can be coupled with 1,4,7,10-tetraazacyclododecane-N,N’,N’’,N’’’-tetraacetic acid (DOTA) as described in Bander's U.S. Patent No. 7,045,605, which is hereby incorporated by reference in its entirety, 111 indium, 90 yttrium, or 177 can be radiolabeled with lutetium.

[0064] In another embodiment of the present invention, the cancer is a neuroendocrine cancer. Neuroendocrine cancers include, but are not limited to, carcinoid tumors, gastrinomas, insulinomas, glucagonomas, VIPomas, somatostatinomas, thyroid cancers, cutaneous Merkel cell cancers, anterior pituitary tumors, medullary cancers, parathyroid tumors, thymic and mediastinal carcinoid tumors, pulmonary neuroendocrine tumors, adrenal medullary tumors, pheochromocytomas, schwannomas, paragangliomas, neuroblastomas, and urothelial carcinoid neuroendocrine cancers.

[0065] According to this aspect of the present invention, in one embodiment, the first and second targeting components target somatostatin receptors.

[0066] At least five somatostatin receptor subtypes have been characterized, and tumors can express various receptor subtypes (Shaer et al., “Somatostatin Receptor Subtypes sst1, sst2, sst3 and sst5 Expression in Human Pituitary, Gastroentero-Pancreatic and Mammary tumors: Comparison of mRNA Analysis With Receptor Autoradiography,” Int. J. Cancer 70:530-537 (1997), which is hereby incorporated by reference in its entirety). Naturally occurring somatostatin and its analogs exhibit different binding to these receptor subtypes, enabling precise targeting of peptide analogs to specific diseased tissues.

[0067] According to this aspect of the invention, the first and second target-directed components have at least one biological activity of native somatostatin, and preferably, this activity is the ability to specifically bind to the somatostatin receptor of cells having a somatostatin receptor. Many such analogs having biological activity are known, for example, U.S. Patent No. 5,770,687 to Hornik et al.; U.S. Patent No. 5,708,135 to Coy et al.; U.S. Patent No. 5,750,499 to Hoeger et al.; U.S. Patent No. 5,620,675 to McBride et al.; U.S. Patent No. 5,633,263 to Coy et al.; U.S. Patent No. 5,597,894 to Coy et al.; U.S. Patent No. 5,073,541 to Taylor et al.; U.S. Patent No. 4,904,642 to Coy et al.; U.S. Patent No. 6,017,509 to Dean; WO98 / 47524 to Hoffman et al.; and U.S. Patent No. 5,411,943 to Bogden, each of which is hereby incorporated by reference in its entirety.

[0068] In one embodiment, the first and second target-directed components target somatostatin receptor-2.

[0069] In another embodiment of the invention, the cancer is breast cancer.

[0070] According to this embodiment of the invention, when the cancer is breast cancer, the first and second target-directed components target the HER receptor family.

[0071] The first and second agents, as well as the target-directed component and the therapeutic component, are as described above.

[0072] In another embodiment of the invention, the cancer is non-Hodgkin lymphoma.

[0073] According to this embodiment, when the cancer is non-Hodgkin lymphoma, the first and second target-directed components target CD20.

[0074] The first and second agents, as well as the target-directed component and the therapeutic component, are as described above.

[0075] Another aspect of the present invention relates to a combination therapy agent for treating cancer. The combination therapy agent includes a first agent including a first target-directed component bound to a first cancer therapeutic component and a second agent including a second target-directed component bound to a second cancer therapeutic component. The first and second target-directed components have different biodistributions and / or pharmacokinetics.

[0076] The first and second agents, as well as the target-directed component and the therapeutic component, are as described above.

[0077] The pharmaceutical composition containing the agent for use in the method of the present invention may include a pharmaceutically acceptable carrier described below, one or more active agents, and a suitable delivery vehicle. Suitable delivery vehicles include, but are not limited to, viruses, bacteria, biodegradable microspheres, microparticles, nanoparticles, liposomes, collagen minipellets, and cochleates.

[0078] In one embodiment of the present invention, a pharmaceutical composition or formulation containing an inhibitory nucleic acid molecule (e.g., an siRNA molecule) is encapsulated within a lipid formulation to form nucleic acid-lipid particles as described in Semple et al., “Rational Design of Cationic Lipids for siRNA Delivery,” Nature Biotech. 28:172-176 (2010), WO2011 / 034798 by Bumcrot et al., WO2009 / 111658 by Bumcrot et al., and WO2010 / 105209 by Bumcrot et al. (which are hereby incorporated by reference in their entirety).

[0079] In another embodiment of the invention, the delivery vehicle is a nanoparticle. A variety of nanoparticle delivery vehicles are known in the art and are suitable for delivery of the inhibitors of the present invention (see, for example, van Vlerken et al., “Multi-functional Polymeric Nanoparticles for Tumour-Targeted Drug Delivery,” Expert Opin. Drug Deliv. 3(2):205-216 (2006), which is incorporated herein by reference in its entirety).Suitable nanoparticles include, but are not limited to, poly(β - amino ester) (Sawicki et al., “Nanoparticle Delivery of Suicide DNA for Epithelial Ovarian Cancer Cell Therapy,” Adv. Exp. Med. Biol. 622:209 - 219 (2008), which is hereby incorporated by reference in its entirety), polyethyleneimine - alt - poly(ethylene glycol) copolymer (Park et al., “Degradable Polyethylenimine - alt - Poly(ethylene glycol) Copolymers As Novel Gene Carriers,” J. Control Release 105(3):367 - 80 (2005), and Park et al., “Intratumoral Administration of Anti - KITENIN shRNA - Loaded PEI - alt - PEG Nanoparticles Suppressed Colon Carcinoma Established Subcutaneously in Mice,” J Nanosci. Nanotechnology 10(5):3280 - 3 (2010), which are hereby incorporated by reference in their entirety), and liposome - encapsulated siRNA nanoparticles (Kenny et al., “Novel Multifunctional Nanoparticle Mediates siRNA Tumor Delivery, Visualization and Therapeutic Tumor Reduction In Vivo,” J. Control Release 149(2):111 - 116 (2011), which is hereby incorporated by reference in its entirety). Other nanoparticle delivery vehicles suitable for use in the present invention include the microcapsule nanotube device disclosed in U.S. Patent Application Publication No. 2010 / 0215724 to Prakash et al. (which is hereby incorporated by reference in its entirety).

[0080] In another embodiment of the present invention, the pharmaceutical composition is contained within a liposomal delivery vehicle. The term "liposome" means a vesicle composed of amphiphilic lipids arranged in a spherical bilayer or bilayer. Liposomes are single - layer or multi - layer vesicles having a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the composition to be delivered. Cationic liposomes have the advantage of being able to fuse with the cell wall. Non - cationic liposomes, although not able to fuse efficiently with the cell wall, are taken up by macrophages in vivo.

[0081] Among the multiple advantages of liposomes are their biocompatibility and biodegradability; the uptake of a wide range of water - and lipid - soluble drugs; and their protection of the encapsulated drugs from metabolism and degradation. Important considerations in the preparation of liposome formulations are the surface charge of the lipids, the size of the vesicles, and the aqueous volume of the liposomes.

[0082] Liposomes are useful for the transfer and delivery of the active ingredient to the site of action. Since the liposomal membrane is structurally similar to the biological membrane, when liposomes are applied to tissue, the liposomes begin to fuse with the cell membrane, and as the fusion of the liposome and the cell progresses, the contents of the liposome flow into the cells where the active agent can act.

[0083] Methods for preparing liposomes for use in the present invention include those disclosed in Bangham et al., “Diffusion of Univalent Ions Across the Lamellae of Swollen Phospholipids,” J. Mol. Biol. 13:238 - 52(1965), U.S. Patent No. 5,653,996 to Hsu, U.S. Patent No. 5,643,599 to Lee et al., U.S. Patent No. 5,885,613 to Holland et al., U.S. Patent No. 5,631,237 to Dzau and Kaneda, and U.S. Patent No. 5,059,421 to Loughrey et al., and these documents are hereby incorporated by reference in their entirety.

[0084] In another embodiment of the present invention, the delivery vehicle is a viral vector. Although viral vectors are particularly suitable for delivering inhibitory nucleic acid molecules such as siRNA and shRNA molecules, they can also be used to deliver molecules encoding anti-integrin antibodies. Suitable gene therapy vectors include, but are not limited to, adenoviral vectors, adeno-associated viral vectors, retroviral vectors, lentiviral vectors, and herpes viral vectors.

[0085] Adenovirus virus vector delivery vehicles can be readily prepared and utilized as described in Berkner, “Development of Adenovirus Vectors for the Expression of Heterologous Genes,” Biotechniques 6:616-627 (1988), Rosenfeld et al., “Adenovirus-Mediated Transfer of a Recombinant Alpha 1-Antitrypsin Gene to the Lung Epithelium In Vivo,” Science 252:431-434 (1991), WO93 / 07283 by Curiel et al., WO93 / 06223 by Perricaudet et al., and WO93 / 07282 by Curiel et al., which are hereby incorporated by reference in their entirety.The adeno-associated virus delivery vehicle can be constructed as described in Shi et al., “Therapeutic Expression of an Anti-Death Receptor-5 Single-Chain Fixed Variable Region Prevents Tumor Growth in Mice,” Cancer Res. 66:11946-53 (2006); Fukuchi et al., “Anti-Aβ Single-Chain Antibody Delivery via Adeno-Associated Virus for Treatment of Alzheimer’s Disease,” Neurobiol. Dis. 23:502-511 (2006); Chatterjee et al., “Dual-Target Inhibition of HIV-1 In Vitro by Means of an Adeno-Associated Virus Antisense Vector,” Science 258:1485-1488 (1992); Ponnazhagan et al., “Suppression of Human Alpha-Globin Gene Expression Mediated by the Recombinant Adeno-Associated Virus 2-Based Antisense Vectors,” J. Exp. Med. 179:733-738 (1994); and Zhou et al., “Adeno-associated Virus 2-Mediated Transduction and Erythroid Cell-Specific Expression of a Human Beta-Globin Gene,” Gene Ther. 3:223-229 (1996), and can be used to deliver the inhibitory nucleic acid molecules of the present invention to cells, and these documents are hereby incorporated by reference in their entirety into this specification.The in vivo use of these vehicles is described in Flotte et al., “Stable in Vivo Expression of the Cystic Fibrosis Transmembrane Conductance Regulator With an Adeno-Associated Virus Vector,” Proc. Nat’l. Acad. Sci. 90:10613-10617 (1993), and Kaplitt et al., “Long-Term Gene Expression and Phenotypic Correction Using Adeno-Associated Virus Vectors in the Mammalian Brain,” Nature Genet. 8:148-153 (1994), which are hereby incorporated by reference in their entirety. Additional types of adenoviral vectors are described in U.S. Patent No. 6,057,155 to Wickham et al.; U.S. Patent No. 6,033,908 to Bout et al.; U.S. Patent No. 6,001,557 to Wilson et al.; U.S. Patent No. 5,994,132 to Chamberlain et al.; U.S. Patent No. 5,981,225 to Kochanek et al.; U.S. Patent No. 5,885,808 to Spooner et al.; and U.S. Patent No. 5,871,727 to Curiel, which are hereby incorporated by reference in their entirety.

[0086] Retroviral vectors modified to form infectious transformation systems can also be used to deliver nucleic acid molecules to target cells. One such type of retroviral vector is disclosed in U.S. Patent No. 5,849,586 to Kriegler et al., which is hereby incorporated by reference in its entirety. Other nucleic acid delivery vehicles suitable for use in the present invention include those disclosed in U.S. Patent Application Publication No. 20070219118 to Lu et al., which is hereby incorporated by reference in its entirety.

[0087] Regardless of the type of infectious transformation system used, this should be aimed at the delivery of nucleic acids to the desired cell type. For example, for delivery to clusters of cells (e.g., cancer cells), a high titer of infectious transformation system can be directly injected into the site of those cells to increase the likelihood of cell infection. The infected cells then express inhibitory nucleic acid molecules aimed at inhibiting integrin expression. The expression system may further include a promoter for controlling or regulating the intensity and specificity of the expression of the nucleic acid molecule in the target tissue or cell.

[0088] The effective dose of the composition of the present invention for the treatment of metastatic diseases varies depending on many different factors such as the type and stage of cancer, the means of administration, the target site, the physiological state of the patient, other drugs or therapies administered, and the physical condition of the patient with respect to other medical complications. The treatment dose needs to be incrementally increased to optimize safety and efficacy.

Example

[0089] The following examples are intended to illustrate the implementation of the embodiments of the present disclosure, but are by no means intended to limit its scope.

[0090] Materials and Methods Internalization analysis. LNCaP and CWR22Rv1 cells were trypsinized and aliquoted into tubes at 1×10 6 cells per tube. The cells were washed once with RPMI (without FBS). Contained in 200 μL of RPMI (without FBS) per tube 177 Lu-J591 (150,000 CPM) and / or 177 Lu-PSMA-617 (110,000 CPM) was added into 200 μL of RPMI (without FBS) per tube. The samples were incubated at 37°C for 1 hour and the cells were resuspended every 15 minutes. After incubation, the unbound / internalized unbound drug was removed by washing the cells twice with 0.1% BSA / PBS. The samples were counted.

[0091] LNCaP xenografts. Each BALB / c nu / nu male mouse was subcutaneously implanted with 5 million LNCaP cells (with Matrigel). Ten days later, the mice were randomly divided into five groups with similar tumor volumes. On day 0, 177 Lu-J591, 177 Lu-PSMA617, or PBS was injected via the tail vein at various doses. Tumor volume and mouse body weight were measured two to three times per week. Using the formula 0.52 * L * L * W, the tumor volume was calculated and plotted against time (days after treatment).

[0092] CWR22Rv1 xenografts. Each BALB / c nu / nu male mouse was subcutaneously implanted with 5 million CWR22Rv1 cells (with Matrigel). Four days later, the mice were divided into five groups to obtain similar tumor volumes. 177 Lu-J591, 177 Lu-PSMA617, or PBS was injected via the tail vein at the indicated doses. Tumor volume and mouse body weight were measured two to three times per week. Using the formula 0.52 * L * L * W, the tumor volume was calculated and plotted against time (days after treatment).

[0093] Example 1 Radiolabeled anti-PSMA antibody J591- 177 Lu and radiolabeled PSMA-617- 177 Lu were co-incubated, and in vitro, additive 177 internalization of Lu occurred in PSMA-positive cells 177 for each agent, J591- 177Lu translocated internally to a similar extent within each of the given cell lines (LNCaP and CWR22Rv1) (Figs. 1A - 1B). The internal translocation was greater in LNCaP than in CWR22Rv1, consistent with higher PSMA expression in the LNCaP cell line. When both agents were co - incubated, both effectively translocated internally simultaneously, and an additive amount of radiolabel was obtained within the cells.

[0094] Example 2 Radiolabeled anti - PSMA antibody J591 - 177 Lu and radiolabeled PSMA - 617 - 177 Combination treatment using Lu results in an additive anti - tumor effect in vivo In LNCaP xenografts, animals treated with PBS (placebo control) developed tumors approximately 1 cm in diameter by day 25 and required euthanasia a few days later. The most excellent anti - tumor effect was achieved with 150 μCi of J591 - 177 Lu (Fig. 2). An equivalent anti - tumor effect was also achieved with half the dose of J591 - 177 Lu (i.e., 75 μCi) and adding a dose of PSMA - 617 - 177 Lu well below the MTD (i.e., 200 μCi) (Fig. 2).

[0095] In CWR22Rv1 xenografts, mice treated with placebo (PBS) required euthanasia by day 17. J591 - 177 Lu at the MTD dose of 180 μCi showed the most excellent anti - tumor response (Fig. 3). This response could be equaled by 135 μCi (75% of the MTD) of J591 - 177 Lu plus 525 μCi (a sub - MTD dose) of PSMA - 617 - 177 Lu (Fig. 3).

[0096] Discussion of Examples 1 - 2 Overall, these data indicate that when two drugs that bind to different sites of a target molecule (e.g., a cell surface receptor molecule) present in a cell population are co-administered, an additive binding of those drugs is obtained. Therefore, when the target molecule / receptor undergoes internalization, an additive amount of the two drugs undergoes internalization. Furthermore, when the two target-directed drugs have different properties (e.g., molecular weight, charge, hydrophilicity / hydrophobicity, pharmacokinetics, biodistribution, etc.) and as a result have different and non-overlapping side effects, the two drugs can be co-administered to bind to / take up target cells additively without causing additional toxicity. As a natural consequence, the dosage of each drug can be appropriately (5 - 50%) reduced below its respective maximum tolerated dose, and thus co-administration of the two target-directed drugs can substantially or completely reduce the toxicity experienced by the subject while delivering an additive dosage to the target cells.

[0097] Predictive Example 1 Administration to Human Patients Tumor-associated molecular targets exemplified by prostate-specific membrane antigen (PSMA) can be targeted in vitro and in vivo by either antibodies or small molecule ligands. These two classes of drugs differ greatly in both molecular size and plasma half-life. TIFF0007701749000001.tif17170 * Containing a linker and a chelate

[0098] As a result of differences in size and their impact on vascular permeability and the ability to invade normal tissue, the biodistribution of these two different classes or types of tumor - targeting agents is different. Low - molecular - weight ligands rapidly exit the blood vessels into the extravascular space and invade into tissues, with little or no significant barriers to entry. In contrast, large - molecular - weight molecules such as antibodies (Ab) circulate within the vascular lumen for days to weeks. These much larger agents can easily pass through reticuloendothelial organs such as the bone marrow and lymph nodes, but other normal tissue barriers, especially in normal tissues where a basement membrane, intervening cell layers, and epithelial tight junctions are present, are traversed only slowly. In the case of invasive tumors, these normal barriers are disrupted by the tumor cells themselves, allowing the Ab to gain easy access to the tumor cells.

[0099] Size is also involved in the length of time a molecule remains in the body before being excreted. For example, small molecules very easily pass through the renal glomeruli and are rapidly excreted in the urine. This applies to small - molecule ligands that target, for example, PSMA or somatostatin type 2 receptor (SSTR - 2). However, since Abs are too large to pass through the glomeruli, they are not excreted via the urinary tract, remain in the body much longer, and are more likely to be metabolized in the liver.

[0100] Two different classes of agents both bind to tumor cells that express their targets (e.g., PSMA, SSTR-2, CD20, etc.), but the normal tissues targeted are different between the classes of agents. For example, PSMA is expressed not only in prostate cancer cells, but also by normal parotid and other salivary glands, lacrimal glands, kidneys, and small intestine (see Figures 4-7). PSMA agents that are predominantly small molecules / inhibitors target these normal tissue sites (parotid and other salivary glands, lacrimal glands, kidneys, and small intestine). Since Abs are too large to penetrate these normal tissues, anti-PSMA Abs do not target the salivary glands or other glands or the kidneys. The different biodistributions of these two types of agents, the large Abs and the small ligands, have been demonstrated by imaging scans of patients administered each type of agent (Figures 8 and 9). When using both types of agents, although the tumor site is targeted, the normal tissues targeted are different and are substantially mutually exclusive.

[0101] By targeting tumors using both of two different types of agents, the tumors receive additive dosing, while the non-targeted normal tissues do not receive additive dosing. Another advantage of targeted delivery using two different types of agents is the option of using more than one type of cytotoxic agent. For example, in radioisotope therapy, α-particles and β-particles or two α-particles or two β-particles can be targeted.

[0102] Predictive Example 2 β+β For example J591-Lu 177 such as β on an Ab (e.g., 177 Lu) targeting PSMA, the key (and dose-limiting) side effect is thrombocytopenia (a decrease in platelet count), and the relative degree of thrombocytopenia gradually increases with increasing dose. None of the patients treated with J591-Lu 177 experienced bleeding symptoms, although some required platelet transfusions until their platelet counts recovered naturally. A dose of 30 mCi / m 2 ×2 (about 2.0 m 2 in males for a total of about 120 mCi) or 35 mCi / m 2At a dose of 2× the dose (total approximately 140 mCi), no patients required platelet transfusions, while at 40 mCi / m 2 ×2 the dose (total approximately 160 mCi) and 45 mCi / m 2 At a dose of 2× the dose (total approximately 180 mCi), 5 out of 16 patients (31%) and 9 out of 15 patients (60%) required platelet transfusions to support them during bone marrow recovery. TIFF0007701749000002.tif28170

[0103] J591-Lu 177 was administered at ≤30 mCi / m given twice at 2-week intervals 2 administered 177 of Lu dose, and began to induce a decrease in PSA as an indicator of antitumor activity. As the dose increased to 140, 160, and 180 mCi [total], the magnitude of the PSA / antitumor response gradually increased.

[0104] Kabasakal et.al., “Pre-Therapeutic Dosimetry of Normal Organs and Tissues of 177 Lu-PSMA-617 Prostate-Specific Membrane Antigen (PSMA) Inhibitor in Patients With Castration-Resistant Prostate Cancer,” Eur.J.Nucl.Med.Mol.Imaging 42:1976 - 1983 (2015) (which is incorporated herein by reference), and Kratochwil et. al., “PSMA-Targeted Radionuclide Therapy of Metastatic Castration-Resistant Prostate Cancer With 177 Lu-Labeled PSMA-617,” J.Nucl.Med. 57:1170 - 1176 (2016) (which is incorporated herein by reference) describe PSMA-617-Lu 177It has been determined that the maximum safe cumulative dose of (representative ligands of PSMA-binding small molecule ligands / inhibitors / peptides) is 27 - 30 Gbq as the limit for the kidneys and salivary glands. At this level or doses above it, patients may develop renal dysfunction and xerostomia (dry mouth). Severe xerostomia leads to loss of taste and appetite and severe dental / oral diseases. Patients complain of difficulty chewing, swallowing, sleeping, talking, dysgeusia, and a burning sensation in the mouth.

[0105] PSMA-617 / Ligand-Lu 177 was administered at a cumulative dose of 300 mCi (11.1 Gbq) in a two-dose regimen (150 mCi × 2) at two-week intervals. 177 The Lu dose begins to induce a decrease in PSA. As the dose is increased to a total of 400 - 500 mCi (14.8 - 18.5 GBq) in the two-dose combination, the magnitude of the PSA / anti-tumor response gradually increases.

[0106] J591- 177 Lu does not deliver a large radiation dose to the kidneys or salivary glands, and PSMA-617 / ligand does not deliver a large dose to the bone marrow, so it does not cause thrombocytopenia. That is, these two different targeted agents both bring about anti-tumor activity, but their respective side effects are mutually exclusive.

[0107] The proposed treatment attempts to achieve the maximum dose to the tumor and the resulting anti-tumor effect by targeting Lu (or other cytotoxic agents known to those skilled in the art) using a combination of two respective agents (anti-PSMA Ab (e.g., J591) and PSMA ligand / inhibitor / peptide (e.g., PSMA-617, PSMA-11, PSMA I&T, RM-2, etc.)). By appropriately reducing the dose of each agent, the side effects on platelets [due to anti-PSMA Ab] and the kidneys and salivary glands [due to PSMA ligand / inhibitor / peptide] can be reduced or avoided. More specifically, a total dose of 120 - 140 mCi of J591- 177 Lu is proposed in a two-week cycle. J591- 177 Lu177 Another possible dose range for Lu is a total of 100 - 160 mCi over a 2 - week cycle. J591 - 177 The administration of Lu should be achieved by co - administration of a PSMA ligand / inhibitor / peptide (e.g., PSMA - 617) at a total of 400 mCi over a 2 - week cycle (or in the range of 300 - 500 mCi over a 2 - week cycle). At these doses, no significant decrease in salivary gland or renal function occurs, and this dose is less than the cumulative dose of 27 - 30 Gbq determined by Kabasakal et.al., “Pre - Therapeutic Dosimetry of Normal Organs and Tissues of 177 Lu - PSMA - 617 Prostate - Specific Membrane Antigen (PSMA) Inhibitor in Patients With Castration - Resistant Prostate Cancer,” Eur.J.Nucl.Med.Mol.Imaging 42:1976 - 1983 (2015) (which is hereby incorporated by reference in its entirety), and Kratochwil et. al., “PSMA - Targeted Radionuclide Therapy of Metastatic Castration - Resistant Prostate Cancer With 177 Lu - Labeled PSMA - 617,” J.Nucl.Med.57:1170 - 1176 (2016) (which is hereby incorporated by reference in its entirety). Optionally, one or two additional doses of 200 - 250 mCi (7.4 - 9.25 Gbq) can be given at intervals of more than 2 weeks, still remaining below the limit of 27 - 30 Gbq.

[0108] Predictive Example 3 α+α Alpha particles offer the potential to kill significantly more cells than beta particles due to their substantially larger atomic mass, linear energy transfer, and radiobiological effects. PSMA - 617 - Ac 225When treating prostate cancer patients using [specific treatment], a significant potential for anti-tumor effects has been reported. Unfortunately, this treatment is limited by unacceptable damage to the salivary glands. Although no toxicity was observed at a treatment activity of 50 kBq / kg, insufficient anti-tumor responses occurred in patients with large tumor volumes. However, when the treatment activity exceeded 100 kBq / kg per cycle, the increase in the administered activity resulted in severe xerostomia, which became a dose-limiting toxicity (Kratochwil et.al., “Targeted Alpha-Therapy of Metastatic Castration-Resistant Prostate Cancer With(225)Ac-PSMA-617:Dosimetry Estimate and Empiric Dose Finding,” J.Nucl.Med. 58(10):1624-1631(2017), which is hereby incorporated by reference in its entirety). Although higher doses resulted in greater anti-tumor activity, the side effects were too large. However, this can be improved by delivering additional Ac 225 to the tumor via an Ab that does not target the salivary glands. The appropriate dose of J591-Ac 225 can be determined in a standard Phase 1 trial where all patients receive incremental doses of J591-Ac 225 in addition to PSMA617-Ac 225 at an acceptable tolerance of 100 Kbq / kg until the MTD is determined. This determines the dose of the combination of PSMA ligand-Ac 225 + anti-PSMA Ab-Ac 225 .

[0109] Predictive Example 4 α+β The combined treatment using α particles, with their high energy, high cytotoxicity, and concentrated short range, along with β particles having a longer range, provides benefits to patients as the latter confer anti-tumor activity at the tumor margin. It is preferred to target α using Ab to avoid unacceptable salivary gland side effects and to target β using a PSMA ligand to avoid severe hematotoxicity. All patients receive PSMA617-Lu 177 at a total of 400 mCi in 2-week cycles (or in the range of 300 - 500 mCi in 2-week cycles). The appropriate dose of J591-Ac 225 can be determined in a standard Phase 1 trial format by escalating doses of J591-Ac 225 in combination with PSMA-617- 177 Lu at the doses described above.

[0110] Alternatively, 177 Lu may be targeted by the Ab and Ac 225 by the PSMA-617. In this case, the latter is administered at 100 kbq / kg and the Ab J591 is administered at a total of J591- 177 Lu of 120 - 140 mCi in 2-week cycles. Another possible dose range for J591- 177 Lu is a total of 100 - 160 mCi in 2-week cycles.

[0111] Although the preferred embodiments have been illustrated and described in detail herein, it will be apparent to those skilled in the art that various modifications, additions, substitutions, etc. can be made without departing from the spirit of the invention. Accordingly, these are considered to be within the scope of the invention as defined in the appended claims.

Claims

1. A therapeutically effective amount of J591- ２２５ Ac and a therapeutically effective amount of a PSMA receptor ligand / inhibitor, a combination therapy agent for treating prostate cancer, comprising The PSMA receptor ligand / inhibitor is selected from the group consisting of PSMA 617, PSMA I&T, DCFBc, DCFPyL, glutamate-urea-lysine, phosphoramidate, and 2-(phosphinylmethyl)pentanedioic acid, and the PSMA receptor ligand / inhibitor is bound to a radionuclide, Combined therapeutic agent.

2. The combined therapeutic agent according to Claim 1, wherein the PSMA receptor ligand / inhibitor is PSMA 617 or PSMA I&T.

3. The combined therapeutic agent according to Claim 2, wherein the PSMA receptor ligand / inhibitor is PSMA 617.

4. The combined therapeutic agent according to Claim 2, wherein the PSMA receptor ligand / inhibitor is PSMA I&T.

5. wherein the radionuclide is selected from one or more of the group consisting of ８６ Re, ９０ Y, ６７ Cu, １６９ Er, １２１ Sn, １２７ Te, １４２ Pr, １４３ Pr, １９８ Au, １９９ Au, １６１ Tb, １０９ Pd, １８８ Rd, １６６ Dy, １６６ Ho, １４９ Pm, １５１ Pm, １５３ Sm, １５９ Gd, １７２ Tm, １６９ Yb, １７５ Yb, １７７ Lu, １０５ Rh, １１１ Ag, １３１ I, １７７ Sn, ２２５ Ac, ２２７ Th, and ２１１ At, and the combined therapeutic agent according to any one of claims 1 to 4.

6. The radionuclide is ２２５ Ac and １７７ The combined therapeutic agent according to claim 5, selected from the group consisting of Lu.

7. The PSMA receptor ligand / inhibitor is １７７ The combined therapeutic agent according to claim 6, which is PSMA 617 conjugated with Lu.

8. The PSMA receptor ligand / inhibitor is １７７ The combined therapeutic agent according to claim 6, which is PSMA I&T bound to Lu.

9. Use of a therapeutically effective amount of J591- ２２５ Ac and a therapeutically effective amount of a PSMA receptor ligand / inhibitor in the manufacture of a combination therapy agent for treating prostate cancer, The PSMA receptor ligand / inhibitor is selected from the group consisting of PSMA 617, PSMA I&T, DCFBc, DCFPyL, glutamate-urea-lysine, phosphoramidate, and 2-(phosphinylmethyl)pentanedioic acid, and the PSMA receptor ligand / inhibitor is bound to a radionuclide, Use.

10. The use according to Claim 9, wherein the PSMA receptor ligand / inhibitor is PSMA 617 or PSMA I&T.

11. The use according to Claim 10, wherein the PSMA receptor ligand / inhibitor is PSMA 617.

12. The use according to Claim 10, wherein the PSMA receptor ligand / inhibitor is PSMA I&T.

13. wherein the radionuclide is ８６ Re, ９０ Y, ６７ Cu, １６９ Er, １２１ Sn, １２７ Te, １４２ Pr, １４３ Pr, １９８ Au, １９９ Au, １６１ Tb, １０９ Pd, １８８ Rd, １６６ Dy, １６６ Ho, １４９ Pm, １５１ Pm, １５３ Sm, １５９ Gd, １７２ Tm, １６９ Yb, １７５ Yb, １７７ Lu, １０５ Rh, １１１ Ag, １３１ I, １７７ Sn, ２２５ Ac, ２２７ Th, and ２１１ At, and the use according to any one of claims 9 to 12, which is selected from one or more of the group consisting of

14. wherein the radionuclide is ２２５ Ac and １７７ Lu, and the use according to claim 13, selected from the group consisting of

15. wherein the PSMA receptor ligand / inhibitor is １７７ the use according to claim 14, wherein the PSMA receptor ligand / inhibitor is PSMA 617 conjugated with Lu.

16. The PSMA receptor ligand / inhibitor is １７７ The use according to claim 14, which is PSMA I&T bound to Lu.

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