Antibody-encapsulated metallofullerene complex and use thereof

Abstract

The present invention relates to endohedral metallofullerene derivatives for use in the treatment of cancer and other proliferative diseases. In particular, the present invention discloses the use of endohedral fullerene antibody conjugates and derivatives thereof for the treatment of proliferative diseases, particularly cancer, including the combined administration of a metallofullerene-antibody conjugate or a composition comprising the same and ionizing radiation therapy, such as X-ray irradiation.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is filed on January 19, 2023 as a PCT international patent application claiming priority to and benefit of U.S. Provisional Patent Application No. 63 / 300,698, filed on January 19, 2022, the disclosure of which is hereby incorporated in its entirety.

[0002] Field Disclosed herein are uses of metallofullerene-antibody conjugates and methods for treating proliferative diseases, particularly cancer.Embodiments include combination therapies that include endohedral metallofullerene-antibody conjugates that selectively target cancer cells and their use for treating cancer in combination with one or more specific ionizing radiation therapies.Embodiments also include kits that include such components and methods for treating proliferative diseases, particularly cancer, with said one or more combination therapies. [Background technology]

[0003] background Normal cells proliferate through the highly regulated activation of growth factor receptors, such as growth factor receptor tyrosine kinases, by their respective ligands.

[0004] Cancer cells also proliferate through the activation of growth factor receptors, but with a loss of normal growth control, which may be caused by a range of factors including autocrine secretion of growth factors, increased expression of receptors, and autonomous activation of biochemical pathways regulated by growth factors.

[0005] Normal cells generally have a single nucleus, a large cytoplasmic volume, controlled growth and generally remain in their intended location, whereas malignant cells, such as cancer cells, have irregular cell shapes and sizes, multiple irregularly shaped nuclei, small cytoplasmic volumes and / or uncontrolled growth, as illustrated, for example, in FIG.

[0006] Cancer is the second leading cause of human death after coronary artery disease. Worldwide, millions of people die from cancer or related diseases every year. In the United States alone, as reported by the American Cancer Society, cancer causes the death of more than 500,000 people every year, with more than 1.2 million new cases diagnosed per year. Deaths from heart disease have decreased significantly, but deaths from cancer are reported to be increasing. Cancer is predicted to be the leading cause of death in the coming decades.

[0007] Globally, several malignancies stand out as the leading lethal cancers. In particular, carcinomas of the lung, prostate, breast, colon, pancreas, and ovary are the leading causes of cancer death. These and almost all other carcinomas share a common lethal factor. Almost without exception, metastatic disease from the carcinoma is fatal. Moreover, even for cancer patients who survive their initial cancer early, a common experience has shown that their lives change dramatically even if they do not have a new recurrence episode. Many cancer patients experience strong mental health effects driven by the awareness of the possibility of recurrence or treatment failure. Many survivors, especially those who experience recurrence, experience physical debilitation after treatment.

[0008] Among cancers, those arising from organs and solid tissues, known as solid cancers, such as colon, lung, breast, gastric, prostate, pancreatic, etc., are among the most commonly identified human cancers.

[0009] In 2005, an estimated 27 million people worldwide suffered from cancer, and 5.1 million deaths were attributed to malignant tumors. There are over 10 million new cases each year, and the number is expected to grow 50% over the next 15 years. Current cancer treatments are generally limited to invasive surgery, radiation therapy, and chemotherapy, each of which is known to have severe side effects, non-specific toxicity, and / or traumatic changes to a person's body image and / or quality of life. Cancer may become resistant to chemotherapy, which may further reduce options and chances of success. Diagnosis for some types of cancer is more poor than others, and some, such as lung or pancreatic cancer, are usually fatal. In addition, certain cancers with relatively high treatment success rates, such as breast cancer, also have very high incidence rates and therefore remain significant causes of death.

[0010] For example, there are over one million new cases of breast cancer worldwide each year. Treatments for treating breast cancer generally include minimal to radical surgical removal of breast tissue and lymph nodes, followed by radiation and chemotherapy. Success rates for localized cancer are relatively good, with a five-year survival rate of approximately 50%. However, for metastatic cancer, which is nearly incurable, a median survival time of approximately two years is expected. Nearly 400,000 women die from breast cancer each year, despite all the improvements in medicines and specific treatments, making it the highest number of cancer deaths in women, surpassing lung cancer deaths.

[0011] Liver cancer is another cancer with a poor prognosis, with more than 500,000 new cases each year and a similar number of deaths due to poor treatment effectiveness. Hepatocellular carcinoma accounts for approximately 80% of all liver cancers and is rarely curable. The 5-year survival rate is only about 10%, and survival after diagnosis is often less than 6 months. Although surgical removal of the diseased tissue can be effective, it is not an option for the majority of cases due to the presence of cirrhosis.

[0012] The American Cancer Society estimates for bladder cancer in the United States in 2021 are approximately 83,730 new cases of bladder cancer (approximately 64,280 in men and 19,450 in women) and approximately 17,200 deaths from bladder cancer (approximately 12,260 in men and 4,940 in women). The rates of new bladder cancer and bladder cancer-related deaths have declined slightly in women in recent years. In men, the incidence has been declining, but mortality has remained stable. Current treatments include intravascular delivery of chemotherapy and immunotherapy with the Bacillus Calmette-Guerin (BCG) vaccine, which carries the added risk of systemic infection with Mycobacterium tuberculosis. Despite this aggressive treatment, 70% of these superficial papillary tumors recur during the long-term clinical course, and some progress to invasive cancer.

[0013] There are many more examples of malignancies where current treatments do not meet the needs of patients due to lack of efficacy and / or because they have high morbidity and severe side effects. The statistics and general data presented herein highlight the need for cancer treatments with better safety and efficacy profiles.

[0014] One of the reasons why current cancer treatments are incomplete is the lack of selectivity for diseased tissues and cells. Surgical removal generally involves the removal of apparently normal tissue as a "safety margin", which may increase the risk of morbidity and complications. It always removes some healthy tissue that may be seeded with tumor cells and may be able to maintain or restore the function of the diseased organ or tissue. Furthermore, traditional radiation and chemotherapy, due to their non-specific mode of action, may kill or damage not only tumor or cancer cells but also many normal cells. This may result in serious side effects such as severe nausea, weight loss and hair loss, as well as an increased risk of developing secondary cancers in the future. A treatment with higher selectivity for cancer cells would leave normal cells intact, thus improving outcomes, side effect profile and quality of life, and is currently one of the biggest challenges in the field.

[0015] The selectivity of cancer therapy can be improved by using antibodies that are specific for molecules present only or primarily on cancer cells. Such antibodies can be used not only to block or modify the function of target molecules, e.g., cancer cells, but also to modulate the immune system and improve the recognition and destruction of cancer by the patient's own immune system. Antibodies may also be used to target drugs, genes, toxins, or other medically appropriate molecules to cancer cells.

[0016] Radiation therapy Radiotherapy or irradiation therapy is the medical use of ionizing radiation, generally as part of cancer treatment to control or kill malignant cells. Radiotherapy may be curative in many types of cancer, and may be used as part of a curative therapy to prevent tumor recurrence after surgery to remove the primary malignant tumor. Radiotherapy is generally used before, during, and after chemotherapy in selected cancers.

[0017] Radiation therapy is commonly applied to cancerous tumors because of its ability to control cell growth. Ionizing radiation works by damaging the DNA of exposed tissues, leading to cell death. Normal tissues (such as the skin or organs through which the radiation must pass to treat the tumor) also receive radiation doses, causing cell damage and death that sometimes lead to serious side effects.

[0018] Oxygen-deprived tumor cells are resistant to radiation and existing chemotherapy techniques. In contrast to cancerous tumors, normal tissues do not have any hypoxic cells. Therefore, if the radiosensitivity of hypoxic cells in tumors is improved by introducing radiosensitizers, radiation therapy for treating cancer will be more effective. Attempts have been made to increase the radiosensitivity of cells using various compounds such as radiosensitizers, but the results have been mixed and there are still many challenges to overcome in order to selectively attack cancer cells while minimizing the side effects of radiation therapy.

[0019] In most cases of patients undergoing radiation therapy, doses of radiation of approximately 40-60 Gy to the target area, delivered in separate doses intended to cause cell death, are considered to be within the acceptable or medical standard. These "high" doses appear to decrease with distance from the target tissue, and some tissues may receive doses that are referred to as "low doses" (100 mGy or less).

[0020] In contrast, diagnostic radiological procedures generally result in smaller doses to target organs. The following table summarizes estimates of the range of effective doses from diagnostic radiological exposure.

[0021] [Table 1]

[0022] In all cases, radiation therapy is known to cause, to varying degrees, undesirable side effects that may adversely affect the patient's life and recovery, especially when high doses need to be administered in one go or within a short period of time.

[0023] Computed tomography Computed tomography (CT) has the advantages of high resolution, low cost and high imaging diagnostic efficiency, and has become one of the most commonly used imaging diagnostic methods in medical practice. CT utilizes the difference in X-ray absorption and penetration by various tissues of the human body to image the examination site to find minor lesions in the body. Currently, most CT contrast agents on the market contain iodine compounds. Iodine has a large atomic number, and most iodine-containing compounds have strong tissue penetration ability, and enhancement in appropriate areas can increase the brightness of imaging diagnostics and improve diagnostic accuracy.

[0024] However, because these commonly used contrast agents lack affinity with tumor tissue, CT can increase the density of tumor lesions through the large amount of tumor blood supply, but only to a limited extent and lacks specificity for tumor lesions, so CT and MRI scans still lack highly reliable parameter indicators for tumor diagnosis. Summary of the Invention [Means for solving the problem]

[0025] For this reason, one of the aims and objectives of the present application may also be to provide an intravenous contrast agent with tumor tissue guiding function for CT contrast scanning and a tumor targeted contrast agent that improves the resolution of tumor lesions by CT scans and potentially their treatment.

[0026] Chemical properties of fullerenes as building blocks of the present invention Fullerenes are closed cage-like molecules composed primarily of sp2 hybridized carbons arranged in hexagons and pentagons. Fullerenes (e.g., C60) were first identified as closed spherical cages produced by condensation from evaporated carbon. Fullerenes are typically classified according to the number of carbon atoms, e.g., C60-fullerene, C70-fullerene, C76-fullerene, C78-fullerene, C80-fullerene, etc. One of the most well-known fullerenes is Buckminsterfullerene (IUPAC name (C60-Ih)[5,6]fullerene), commonly referred to as "C60" or "Buckyball", as illustrated in FIG. 2. It is well understood that the antibody-endohedral fullerene conjugate may include an endohedral fullerene containing two or more sp3 hybridized carbons upon conjugation.

[0027] Fullerenes and their derivatives are useful as superconductors, catalysts and nonlinear optical materials. In the biomedical field, fullerene compounds find utility as molecular carriers for drugs or catalysts, as carriers of radioactive metals and as radionuclide tracers in targeted therapy for cancer.

[0028] C60 fullerene is a very powerful antioxidant with an antioxidant power 125 times that of vitamin C. In addition to its antioxidant properties, C60 fullerene also has functions such as scavenging free radicals and revitalizing skin cells (preventing death). Since 1990, great progress has been made in the research into the free radical scavenging function of C60 fullerene. Many scientific research results have demonstrated that C60 fullerene is a very powerful free radical scavenging molecule, which has the potential to generate significant developments in the research of new therapies for cancer treatment.

[0029] Endohedral fullerene Endohedral metallofullerenes (EMFs) are fullerenes that encapsulate metals or metal clusters. Endohedral metallofullerenes are an interesting class of fullerenes because electron transfer from the trapped metal atom to the carbon cage is known to occur, which often dramatically modifies the electrical and magnetic properties of the fullerene. Since the discovery of fullerenes, many attempts to encapsulate various components within the carbon structure have resulted in the provision of compounds with remarkable structural, electrical and chemical properties suitable for many applications in medical and biological diagnostics and therapeutics.

[0030] Endohedral metallofullerenes (EMFs) are particularly attractive nanoparticles due to their shape, capacity for various encapsulating agents, and good isolation from the biological environment, making them an ideal nanoplatform for designing next-generation diagnostic and therapeutic biomedical agents. These nanoparticles can easily undergo extravasation from the blood pool into tumor tissue and can be retained due to poor lymphatic drainage. This phenomenon of selective accumulation of nano-sized particles near tumor tissue is called the Enhanced Vascular Permeability Retention (or EPR) effect, leading to the accumulation of nanoparticles near tumor tissue. Recent successful production and isolation of endohedral metallofullerenes has facilitated their chemical functionalization, which facilitates the clarification of their physical and chemical properties.

[0031] The therapeutic potential of EMF has also been an area of ​​great interest. Numerous studies have demonstrated the free radical scavenging ability of fullerenes, to the point that some authors have described them as "free radical sponges". In addition, antitumor and radical scavenging properties have been reported for a few metallofullerenes, such as the Gd@C82 platform.

[0032] Due to the severity and prevalence of proliferative diseases, including tumors and cancers, there is a great need for effective pharmaceutical compositions, uses and methods for the treatment of such diseases that overcome the shortcomings of surgery, chemotherapy and radiation therapy. Fullerenes, particularly the endohedral fullerenes described herein, are suitable candidates for the development of effective new methods for the treatment of proliferative diseases, including cancer. [Brief description of the drawings]

[0033] [Figure 1] FIG. 1 shows a schematic diagram of normal cells (1) versus cancer or malignant (2) cells. [Diagram 2] FIG. 2 shows a schematic diagram of the chemical structure of a fullerene C60-cage. [Diagram 3] FIG. 3 shows a schematic diagram of an endohedral fullerene exposed to ionizing radiation. [Figure 4] FIG. 4 shows a schematic diagram of a normal cell (1), a normal membrane receptor (1a), a malignant cell (2), an altered membrane receptor (3), and the binding of the altered membrane receptor (3) to a metallofullerene-antibody complex (5). [Diagram 5] FIG. 5 shows a schematic diagram of the binding of an engineered membrane receptor (3) to a metallofullerene-antibody complex (5) subjected to low levels of ionizing radiation (6). [Figure 6] FIG. 6 shows a schematic diagram of the binding of an engineered membrane receptor (3) to a cell-binding moiety (4) of a metallofullerene-antibody conjugate (5) containing two fullerene moieties, subjected to low levels of radiation (6). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Detailed Description Reference will now be made in detail to certain embodiments. While enumerated embodiments will be described, it will be understood that they are not intended to limit the invention to those embodiments. To the contrary, the invention is intended to cover alternatives, modifications, and equivalents, which may be included within the scope of the invention as defined by the claims. Those skilled in the art will recognize several methods equivalent to those described herein, which may be used in the practice of the invention. In the event that one or more of the incorporated literature and similar materials differs or is inconsistent with this application, including but not limited to defined terms, term usage, techniques described, or the like, this application will control.

[0035] To solve the above-mentioned problems in the prior art, it is an object of the present disclosure to provide endohedral metallofullerene derivatives that are therapeutically activated by ionizing radiation for use in the treatment of proliferative diseases, including cancer, and therapeutic methods for the treatment of tumors and cancer.

[0036] The present invention is based on the discovery that by administering to a patient a monoclonal or polyclonal antibody-encapsulated metallofullerene complex or a composition comprising the same, which targets an altered receptor or membrane of a particular cancer cell, where the metal is a rare earth element, and subsequently applying a source of low intensity ionizing radiation, such as X-ray irradiation, to the patient, particularly to the area where the tumor is located, it is possible to target said cancer cells in situ, the treatment is highly effective, damages the genetic material of the cell, its DNA, its membrane, receptors or a combination thereof, requires a low dose of radiation, and subsequently causes little to no undesirable side effects.

[0037] In particular, it has been surprisingly found that the reactivity of rare earth metal elements to ionizing radiation may be exponentially increased by embedding the metal element within a fullerene, thus forming an endohedral fullerene. The reactivity of fullerenes, such as 60-, 80- or 82-carbon fullerene derivatives encapsulating rare earth lanthanide metals, to X-ray irradiation has shown promising results. When lanthanide endohedral fullerenes are subjected to X-ray irradiation, for example at 160 electron volts, a reaction occurs that induces the lanthanide element to lose an electron, which transitions to a lower level atom. The vacant electron site of the transition is then occupied by an outer shell electron from the rare earth lanthanide element, and the energy produced by such an electronic transition is absorbed by the carbon fullerene cage or by another lanthanide atom. This phenomenon, also referred to herein as electron transfer chain reaction, facilitates the induction of radiation-induced cell damage by exposing molecules to low X-ray radiation, which may enhance the effect of such low X-ray radiation based on the energy absorption produced by the fullerene cage, producing highly specific localized and targeted cell damage. Furthermore, the electron transfer chain reaction described herein allows for in situ cell damage in all malignant cells surrounding or located near the metallofullerene-antibody complex, which increases the selectivity of the treatment and reduces damage to healthy normal cells. The metallofullerene-antibody complex of the present invention therefore has the potential to act as both an acceptor and an enhancer of ionizing radiation.

[0038] The present invention provides endohedral metallofullerene-antibody conjugates and compositions comprising said conjugates for use in the treatment of proliferative diseases, particularly cancer. The present invention further describes a method comprising administering an endohedral metallofullerene-antibody conjugate to a patient in combination with low-level ionizing radiation, such as X-ray irradiation. The endohedral metallofullerene-antibody conjugates and methods of the present invention have surprisingly been shown to reduce the need for exposure to said radiation in patients while achieving high therapeutic efficacy. In another embodiment, the present invention provides compositions comprising endohedral metallofullerenes and their use as tumor-targeting imaging agents.

[0039] The present invention represents a breakthrough in the state of the art treatment of cancer and other proliferative diseases and has advantages including:

[0040] Highly specific targeting of malignant (cancer) cells The in situ cell damage in cancer cells achieved by the present invention is facilitated by administration of metallofullerene-antibody conjugates that target and accumulate in close proximity to specific malignant cells. As explained above, the metallofullerene-antibody conjugates act as both an acceptor and an enhancer of ionizing radiation, and such targeted administration of the metallofullerene-antibody conjugates increases the specificity of the ionizing radiation to target and kill only malignant cells, thus reducing damage to healthy normal cells, in sharp contrast to that typically seen in conventional radiation therapy.

[0041] Low levels of ionizing radiation As a result of the increased specificity of the methods of the present invention, radiation doses can be significantly reduced while maintaining or even improving the efficacy of ionizing radiation and at the same time reducing its radiotoxicity. As explained in the detailed description, the present invention has the potential to reduce the amount of ionizing radiation required to destroy malignant cells compared to conventional treatments.

[0042] A wide range of cancer cell types Given that the endohedral metallofullerene-antibody conjugates of the present invention may include one or more antibody carriers that bind to various growth factors, receptors, or membranes, the methods of the present invention have the ability to target and kill a wide range of malignant cancers, including lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, colon / colorectal cancer, bladder cancer, thyroid cancer, breast cancer, liver cancer, ovarian cancer, endometrial cancer, cervical cancer cells, kidney cancer, brain cancer, or melanoma cells. In some cases, the tumor cells can be non-small cell lung cancer (NSCLC) cells.

[0043] In certain embodiments of the present invention, metallofullerene-antibody conjugates are described in which the fullerene is an endohedral metallofullerene, which are useful in the treatment of proliferative diseases including cancer.

[0044] In accordance with the present invention, an endohedral metallofullerene-antibody conjugate for use in the treatment of proliferative diseases in combination with low levels of ionizing radiation has the formula (I): AbC m M n @F 2v (I) (Ab indicates cell-binding agent; M represents a rare earth element in the periodic table of the elements selected from scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, lutetium, or combinations thereof; F represents a fullerene having v carbon atoms; C represents a linker that chemically bonds fullerene F to antibody A; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer of about 20 to about 60. and wherein the metallofullerene-antibody conjugate is therapeutically activated by ionizing radiation.

[0045] In one aspect of the invention, the Ab is a cell binding agent selected from a chimeric, CDR-grafted, humanized or recombinant human antibody. Preferably, the Ab specifically binds to a tumor-associated antigen or cell surface receptor. In another aspect, the Ab is selected from the group of antibodies or antibody fragments used in oncology that target: HER2, CD52, VEGF (vascular endothelial growth factor), EGFR (epidermal growth factor receptor), CD11a, CCR4 (chemokine CC receptor 4), CD105, CD123, CD137, CD19, CD22, CD23, CD3, CD30, CD38, CD4, CD40, CD55SC-1, CD56, CD6, CD74, CD80 , CS1 (cell surface glycoprotein 1), CTLA4 (cytotoxic T-lymphocyte antigen 4, also known as CD152), DR5 (death receptor 5), Ep-CAM (epithelial cell adhesion molecule), folate receptor alpha, ganglioside GD2, ganglioside GD3, GPNMB, glycoprotein NMB, HGF / SF (hepatocyte growth factor / scatter factor), IGF-1 (insulin-like growth factor), IGF1-receptor (insulin-like growth factor-1 receptor), IL13 (insulin-like growth factor-1 receptor), interleukin-13), IL6 (interleukin-6), IL-6R (interleukin-6 receptor), immunodominant fungal antigen, heat shock protein 90 (hsp90), integrin alpha 5 beta 3, MHC (major histocompatibility complex) class II, MN-antigen (G250 antigen), MUC1, PD-1 (programmed death 1), PIGF (placental growth factor), PDGFRa (platelet-derived growth factor receptor alpha), prostate-specific membrane antigen (PSMA), PTHrP (parathyroid hormone-related protein), CD200 receptor, receptor activator of nuclear factor kappa B ligand (RANKL), sphingosine-1-phosphate (SIP), TGF beta, TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) receptor 1, tumor necrosis factor receptor 2, vascular endothelial growth factor receptor 2 (VEGFR-2), CD33, CD20, CA125 (cancer antigen 125) or epidermal growth factor receptor.

[0046] In another embodiment, the antibody of the metallofullerene-antibody conjugate is selected from HER2 (Trastuzumab) - a therapeutic monoclonal antibody specific for human epidermal growth factor receptor type 2 (HER2), a cell surface tyrosine kinase receptor overexpressed by 25% to 30% of breast cancers. In another embodiment, the antibody of the metallofullerene-antibody conjugate is selected from monoclonal antibodies specific for EGFR (Depatuxizumab) - overexpression has been associated with many cancers, and overexpression (also known as upregulation or amplification) has been associated with many cancers, including adenocarcinoma of the lung (40% of cases), anal carcinoma, glioblastoma (50%), and epithelial tumors of the head and neck. In another embodiment, the antibody of the metallofullerene-antibody conjugate is selected from VEGF (Valisacumab) - an antibody specific for vascular endothelial growth factor, overexpression of which may be an early step in the process of metastasis.

[0047] In another embodiment, M represents a rare earth element in the periodic table of the elements, preferably selected from holmium, lutetium, scandium, gadolinium, and combinations thereof.

[0048] In one embodiment, the antibody is, for example, Ab[CM n @F 2v ,CM n @ 2v The complex may contain multiple, such as two or three, endohedral metallofullerenes to provide a complex shown as:

[0049] In another embodiment, m is 0 (zero), in which case the Ab and M components are directly linked.

[0050] The endohedral metallofullerenes of formula (I) and their respective antibody conjugates may be prepared according to known procedures, see, for example, Bolskar, Chen, Shultz and Yang.

[0051] In yet another aspect, the present invention provides a pharmaceutical composition comprising an effective amount of a metallofullerene-antibody conjugate of formula (I) for use in the treatment of a proliferative disease in combination with low levels of ionizing radiation and a pharma- ceutically acceptable carrier or vehicle. Preferably, for use in the treatment of a proliferative disease in combination with low levels of ionizing radiation, the metallofullerene-antibody conjugate is therapeutically activated by ionizing radiation.

[0052] In yet another aspect, the present invention provides the use of an endohedral metallofullerene-antibody conjugate for the treatment of a proliferation-related disorder or disease. The present invention also provides the use of a metallofullerene-antibody conjugate in combination with low levels of ionizing radiation for the treatment of a proliferation-related disorder or disease. In yet another aspect, the present invention provides the use of a metallofullerene-antibody conjugate that is therapeutically activated through ionizing radiation, such as X-rays, for the treatment of a proliferation-related disorder or disease.

[0053] In another aspect, the present invention includes a method for treating cancer, comprising administering to a human having a hyperproliferative disorder a composition comprising a metallofullerene-antibody conjugate and a pharma- ceutically acceptable diluent, carrier or excipient. In another aspect, the present invention provides a method for preventing and / or treating tumor or cancer cell proliferation, comprising administering to a mammal, such as a patient having a hyperproliferative disorder, an effective amount of a metallofullerene-antibody conjugate in combination with low levels of ionizing radiation for the treatment of a proliferation-related disorder or disease, preferably cancer.

[0054] In another aspect, the present invention includes a pharmaceutical composition comprising an effective amount of an endohedral metallofullerene-antibody conjugate of formula (I) or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable diluent, carrier, or excipient. The composition may further comprise a therapeutically effective amount of a chemotherapeutic agent, provided in combination with low levels of ionizing radiation for the treatment of a proliferation-related disorder or disease.

[0055] The pharmaceutical composition is in a form suitable for administration orally, intravenously, topically, by inhalation or via suppository.

[0056] The method according to the invention may comprise the steps of providing to the tumor area an endohedral metallofullerene-antibody conjugate of formula (I) or a composition comprising the conjugate and exposing the treated tumor area to ionizing radiation as defined herein.

[0057] The method according to the present invention may also include the steps of treating the tumor area with a metallofullerene-antibody conjugate of formula (I), concentrating the conjugate or a component thereof in or around the tumor area, and exposing the treated tumor area to ionizing radiation as defined herein.

[0058] Administration of a metallofullerene-antibody conjugate of formula (I) and ionizing radiation, preferably low intensity radiation, has been shown to produce a synergistic effect, such as an antitumor effect, that is greater than the combined effect achieved by either the metallofullerene-antibody conjugate of formula (I), its components, and ionizing radiation individually.

[0059] In a further aspect, the present invention discloses the use of an endohedral metallofullerene-antibody conjugate of formula (I) for use in the preparation of a CT imaging agent.

[0060] In the present invention, the term "ionizing radiation" refers to radiation that contains particles or photons with sufficient energy or generates sufficient energy through interaction with atomic nuclei to cause ionization (gain or loss of electrons). An exemplary and preferred ionizing radiation is X-ray radiation, which is preferably low intensity radiation. Means for delivering X-ray radiation to target tissues or cells are well known in the art. The amount of ionizing radiation required in a given cell generally depends on the nature of the cell, but is generally lower than that required by traditional radiation therapy. As used herein, the term "effective dose" of ionizing radiation refers to the dose of ionizing radiation that causes an increase in cell damage or death when given simultaneously with the endohedral fullerene complex of the present invention and optionally further combined with a chemotherapeutic agent. Effective doses include from about 0.02 mGy to about 60 Gy, as well as, for example, 0.04 mGy, 0.06 mGy, 0.08 mGy, 0.1 mGy, 0.2 mGy, 0.4 mGy, 0.6 mGy, 0.8 mGy, 1 mGy, 2 mGy, 4 mGy, 6 mGy, 8 mGy, 10 mGy, 20 mGy, 40 mGy, 60 mGy, 80 mGy, 100 mGy, 200 mGy, 400 mGy, 600 mGy, 800 mGy, 1000 mGy (or may include, but is not limited to, doses of 1 Gy), 2 Gy, 4 Gy, 6 Gy, 8 Gy, 10 Gy, 12 Gy, 14 Gy, 16 Gy, 18 Gy, 20 Gy, 22 Gy, 24 Gy, 26 Gy, 28 Gy, 30 Gy, 32 Gy, 34 Gy, 36 Gy, 38 Gy, 40 Gy, 42 Gy, 44 Gy, 46 Gy, 48 Gy, 50 Gy, 52 Gy, 54 Gy, 56 Gy, and 58 Gy, and all values ​​therebetween.

[0061] The phrase "pharmacologically acceptable" indicates that the substance or composition is chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal being treated therewith.

[0062] The phrase "therapeutically effective amount" or "effective amount" means an amount of a compound described herein that, when administered to a mammal in need of such treatment, is sufficient to (i) treat or prevent a particular disease, condition or disorder, (ii) attenuate, ameliorate or eliminate one or more symptoms of a particular disease, condition or disorder, or (iii) prevent or delay the onset of one or more symptoms of a particular disease, condition or disorder described herein. The amount of compound corresponding to such an amount will vary depending on factors such as the particular compound, the disease state and its severity, the personality (e.g., weight) of the person in need of treatment, but can nevertheless be routinely determined by one of skill in the art.

[0063] The term "treat" or "treatment" refers to therapeutic, prophylactic, symptomatic or preventative measures. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of the extent of disease, a stable (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation and remission (whether partial or total) of a disease state, whether detectable or undetectable. "Treatment" can also mean prolonging survival as compared to expected survival in the absence of treatment. Those in need of treatment include those already with a condition or disorder as well as those prone to have the condition or disorder or those in which the condition or disorder is to be prevented.

[0064] The amount of endohedral metallofullerene-antibody conjugate for use in a given treatment (e.g., treatment of a proliferative disorder (e.g., cancer) or CT scanning) may be determined. For example, HERCEPTIN® (Trastuzumab) for injection is generally prescribed for the treatment of adjuvant, metastatic and metastatic breast cancer according to a specific dosage and schedule. HERCEPTIN® PI. The amount of endohedral metallofullerene-antibody conjugate disclosed herein may be dosed in the same amount as for HERCEPTIN®, e.g., 2 mg / kg, 4 mg / kg, 6 mg / kg and / or 8 mg / kg. The dosing schedule may be associated with the administration of low levels of ionizing radiation as disclosed herein to minimize tumor growth and / or size. With the foregoing in mind, an effective amount of endohedral metallofullerene-antibody conjugate ranges from about 0.001 mg / kg to about 100 mg / kg and all values ​​therebetween.

[0065] The term "about" is used herein to mean approximately, in the region of, roughly, or approximately. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the limits above and below the numerical values ​​set forth. In general, the term "about" is used herein to modify numerical values ​​above and below the stated value by a variance of 20%.

[0066] Aspects Embodiment 1. A compound of formula (I) for use in the treatment of a proliferative disease in combination with low levels of ionizing radiation: AbC m M n @F 2v (I) (Ab indicates cell-binding agent; M represents a rare earth element in the Periodic Table of the Elements selected from the group including scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F with Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer of about 20 to about 60. The endohedral metallofullerene-antibody complex is The metallofullerene-antibody conjugate is optionally an endohedral metallofullerene-antibody conjugate that is therapeutically activated by ionizing radiation.

[0067] Aspect 2. The metallofullerene-antibody conjugate of aspect 1, wherein the Ab is a cell binding agent selected from the group comprising chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably, the Ab specifically binds to a tumor-associated antigen or cell surface receptor.

[0068] Embodiment 3. Abs include: HER2, HER2, HER3, HER4 CD52, VEGF (vascular endothelial growth factor), EGFR (epidermal growth factor receptor), CD11a, CCR4 (chemokine CC receptor 4), CD105, CD123, CD137, CD19, CD22, CD23, CD3, CD30, CD38, CD4, CD40, CD55SC-1, CD56, CD6, CD74, CD80, CS1 (cell surface glycoprotein 1), CTLA4 (cytotoxic T lymphocyte antigen 4, also known as CD152), DR5 (death receptor 5), Ep -CAM (epithelial cell adhesion molecule), folate receptor alpha, ganglioside GD2, ganglioside GD3, GPNMB, glycoprotein NMB, HGF / SF (hepatocyte growth factor / scatter factor), IGF-1 (insulin-like growth factor), IGF1-receptor (insulin-like growth factor-1 receptor), IL13 (interleukin-13), IL6 (interleukin-6), IL-6R (interleukin-6 receptor), immunodominant fungal antigens, heat shock proteins 3. The metallofullerene-antibody conjugate of any one of embodiments 1-2, wherein the metallofullerene-antibody conjugate is selected from the group of antibodies or antibody fragments used in oncology targeting hsp90, integrin alpha 5 beta 3, MHC (major histocompatibility complex) class II, MN-antigen (also known as G250 antigen), MUC1, PD-1 (programmed death 1), PIGF (placental growth factor), PDGFRa (platelet-derived growth factor receptor alpha), prostate-specific membrane antigen (PSMA), PTHrP (parathyroid hormone-related protein), CD200 receptor, receptor activator of nuclear factor kappa B ligand (RANKL), sphingosine-1-phosphate (SIP), TGF beta (transforming growth factor beta), TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) receptor 1, tumor necrosis factor receptor 2, vascular endothelial growth factor receptor 2 (VEGFR-2), CD33, CD20, CA125 (cancer antigen 125) or epidermal growth factor receptor.

[0069] Embodiment 4. The metallofullerene-antibody conjugate of embodiment 2, wherein the antibody is selected from one or more of EGFR, HER2, HER3 and HER4 binding antibodies.

[0070] Embodiment 5. The metallofullerene-antibody conjugate of embodiment 2, wherein the antibody is selected from one or more of depatuxizumab, valisacumab, and trastuzumab.

[0071] Aspect 6. The metallofullerene-antibody conjugate of any one of Aspects 1-5, wherein M is selected from one or more of holmium, lutetium, and scandium.

[0072] Embodiment 7. The metallofullerene-antibody conjugate of any one of Embodiments 1-6, wherein the proliferative disease is cancer.

[0073] Embodiment 8. The metallofullerene-antibody conjugate of embodiment 7, wherein the type of cancer is selected from one or more of lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, colon / colorectal cancer, bladder cancer, thyroid cancer, breast cancer cells, liver cancer, ovarian cancer, endometrial cancer, cervical cancer cells, kidney cancer, brain cancer, and melanoma.

[0074] Embodiment 9. The metallofullerene-antibody conjugate of any one of Embodiments 1-8, wherein the low level ionizing radiation ranges from about 0.02 mGy to about 60 Gy.

[0075] Aspect 10. A pharmaceutical composition comprising the metallofullerene-antibody conjugate of any one of Aspects 1-8 and a pharma- ceutically acceptable excipient, diluent, or vehicle.

[0076] Aspect 11. The composition of aspect 10 in a form suitable for oral administration, intravenous administration, topical administration, administration by inhalation, or administration via a suppository.

[0077] Embodiment 12. The metallofullerene-antibody conjugate of any one of embodiments 1-9 or the pharmaceutical composition of any one of embodiments 10-11, further comprising an additional therapeutic agent.

[0078] Embodiment 13. A method for treating a cell proliferation associated disorder or disease, comprising: Formula (I): AbC m M n @F 2v (I) (Ab indicates cell-binding agent; M represents a rare earth element in the periodic table of the elements selected from scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F with Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; v represents an integer of about 20 to about 60. administering to the patient an effective amount of a metallofullerene-antibody complex; and A method comprising administering a low level of ionizing radiation, wherein the metallofullerene-antibody complex is preferably therapeutically activated by the ionizing radiation.

[0079] Embodiment 14. The method of embodiment 13, wherein the Ab is a cell binding agent selected from the group comprising chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably the Ab specifically binds to a tumor-associated antigen or a cell surface receptor.

[0080] Aspect 15. Abs include: HER2, HER2, HER3, HER4 CD52, VEGF (vascular endothelial growth factor), EGFR (epidermal growth factor receptor), CD11a, CCR4 (chemokine CC receptor 4), CD105, CD123, CD137, CD19, CD22, CD23, CD3, CD30, CD38, CD4, CD40, CD55SC-1, CD56, CD6, CD74, CD80, CS1 (cell surface glycoprotein 1), CTLA4 (cytotoxic T lymphocyte antigen 4, also known as CD152), DR5 (death receptor 5) , Ep-CAM (epithelial cell adhesion molecule), folate receptor alpha, ganglioside GD2, ganglioside GD3, GPNMB, glycoprotein NMB, HGF / SF (hepatocyte growth factor / scatter factor), IGF-1 (insulin-like growth factor), IGF1-receptor (insulin-like growth factor-1 receptor), IL13 (interleukin-13), IL6 (interleukin-6), IL-6R (interleukin-6 receptor), immunodominant fungal antigens, heat shock 15. The method of any one of aspects 13-14, wherein the antibody or antibody fragment used in oncology is selected from the group of antibodies or antibody fragments used in oncology targeting protein 90 (hsp90), integrin alpha 5 beta 3, MHC (major histocompatibility complex) class II, MN-antigen (also known as G250 antigen), MUC1, PD-1 (programmed death 1), PIGF (placental growth factor), PDGFRa (platelet-derived growth factor receptor alpha), prostate-specific membrane antigen (PSMA), PTHrP (parathyroid hormone-related protein), CD200 receptor, receptor activator of nuclear factor kappa B ligand (RANKL), sphingosine-1-phosphate (SIP), TGF beta (transforming growth factor beta), TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) receptor 1, tumor necrosis factor receptor 2, vascular endothelial growth factor receptor 2 (VEGFR-2), CD33, CD20, CA125 (cancer antigen 125) or epidermal growth factor receptor.

[0081] Embodiment 16 The method of embodiment 14, wherein the antibody is selected from one or more of EGFR, HER2, HER3, and HER4 binding antibodies.

[0082] Embodiment 17 The method of embodiment 14, wherein the antibody is selected from one or more of depatuxizumab, valisacumab, and trastuzumab.

[0083] Embodiment 18. The method of any one of embodiments 13-17, wherein M is selected from one or more of holmium, lutetium, and scandium.

[0084] Embodiment 19. The method of any one of embodiments 13 to 18, wherein the proliferative disease is cancer.

[0085] Embodiment 20. The method of embodiment 19, wherein the type of cancer is selected from one or more of lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, colon / colorectal cancer, bladder cancer, thyroid cancer, breast cancer cells, liver cancer, ovarian cancer, endometrial cancer, cervical cancer cells, kidney cancer, brain cancer, and melanoma.

[0086] Embodiment 21. The method of any one of embodiments 13 to 20, wherein the low level ionizing radiation ranges from about 0.02 mGy to about 60 Gy.

[0087] Embodiment 22 The method of any one of embodiments 13 to 21, wherein the complex is comprised in a pharma- ceutically acceptable composition comprising an excipient, diluent or vehicle.

[0088] Embodiment 23. The method of embodiment 22, wherein the pharmaceutical composition is in a form suitable for administration orally, intravenously, topically, by inhalation, or via a suppository.

[0089] Embodiment 24. The method of any one of embodiments 13-23, further comprising administering one or more additional therapeutic agents.

[0090] Embodiment 25 The method of any one of embodiments 13 to 24, wherein the metallofullerene-antibody complex is administered in the form of an intravenous infusion.

[0091] Embodiment 26 The method of any one of embodiments 13 to 25, wherein the conjugate is administered to the patient as a first line therapy.

[0092] Embodiment 27 The method of any one of embodiments 13 to 25, wherein said conjugate is administered to the patient as a second, third, fourth, fifth or sixth line treatment.

[0093] Embodiment 28 The method of any one of embodiments 13 to 27, wherein the conjugate is administered to the patient after at least one treatment with an anti-cancer therapy.

[0094] Embodiment 29 The method of any one of embodiments 13 to 28, wherein said cell proliferation-related disorder or disease is resistant to at least one anti-cancer agent.

[0095] Aspect 30. The method of any one of aspects 13 to 29, wherein said method for treating a cell proliferation associated disorder or disease inhibits metastasis in said patient.

[0096] Embodiment 31 The method of any one of embodiments 13 to 30, wherein the conjugate is administered to a patient, and once the conjugate has concentrated in target cells, a low level of ionizing radiation is subsequently administered.

[0097] Embodiment 32. The method of any one of embodiments 13 to 31, wherein the low level ionizing radiation is administered to the patient within 24 hours, preferably within 12 hours, and more preferably within 6 hours of administration of the conjugate.

[0098] Aspect 33. Formula (I): AbC m M n @F 2v (I) (Ab indicates cell-binding agent; M represents a rare earth element in the periodic table of the elements selected from scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F with Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer of about 20 to about 60. US20100233633A1 - Endohedral metallofullerene-antibody conjugates for use in the preparation of computed tomography contrast agents.

[0099] Aspects 2 to 12 are equally applicable to aspect 33. For example, aspect 2 specifies that the Ab is a cell binding agent selected from the group comprising chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably the Ab specifically binds to a tumor-associated antigen or a cell surface receptor. Aspect 2 is equally applicable to aspect 33 and any one of aspects 3 to 12.

[0100] Embodiment 34. A method for imaging a tumor in a patient, comprising: administering to the patient an effective amount of the endohedral metallofullerene-antibody conjugate of embodiment 33; and The method includes exposing tissue of a patient to low levels of ionizing radiation.

[0101] Aspects 2 to 12 are equally applicable to aspect 34. For example, aspect 2 specifies that the Ab is a cell binding agent selected from the group comprising chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably the Ab specifically binds to a tumor-associated antigen or a cell surface receptor. Aspect 2 is equally applicable to aspect 34 and any one of aspects 3 to 12.

[0102] Disclosed Information Chen et al., Applications of functionalized fullerenes in tumor theranostics, Theranostics (2012) 2:238-250 (“Chen”).

[0103] HERCEPTIN® (trastuzumab) for injection prescribing information as of November 29, 2018 ("HERCEPTIN® PI").

[0104] Shultz et al.,Encapsulation of a radiolabeled cluster inside a fullerene cage 177 Lu x Lu (3-x) N@C 80 :an Interleukin-13-conjugated radiolabeled metallofullerene platform, J.Am.Chem.Soc.(2010)132(14):4980-4981.

[0105] No. 7,208,132 B2, issued April 24, 2007 to Bolskar et al., to TDA Research, Inc., Purification of endohedral and other fullerenes by chemical methods, and U.S. Patent No. 7,671,230 B2, issued March 2, 2010 to Bolskar et al., to TDA Research, Inc., Bolskar et al., Derivation and solubilization of insoluble classes of fullerenes (collectively, "Bolskar").

[0106] Yang et al., Fullerene-biomolecule conjugates and their biomedical applications, International Journal of Nanomedicine (2014) 9:77-92 (“Yang”).

[0107] The subject matter of U.S. Provisional Patent Application No. 63 / 300,698, filed January 19, 2022, is incorporated by reference. To the extent necessary, the information disclosed herein is hereby incorporated by reference. In the event that the incorporated subject matter conflicts with the subject matter disclosed herein, the subject matter disclosed herein controls.

Claims

1. 4. A compound of formula (I): AbC ｍ M ｎ @F ２ｖ (I) (Ab indicates cell binding agent; M represents a rare earth element in the Periodic Table of the Elements selected from the group including scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F to said Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer from about 20 to about 60. The endohedral metallofullerene-antibody complex is An endohedral metallofullerene-antibody conjugate, wherein said metallofullerene-antibody conjugate is optionally therapeutically activated by said ionizing radiation.

2. The metallofullerene-antibody complex of claim 1, wherein the Ab is a cell binding agent selected from the group including chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably, the Ab specifically binds to a tumor-associated antigen or a cell surface receptor.

3. Abs include: HER2, HER2, HER3, HER4 CD52, VEGF (vascular endothelial growth factor), EGFR (epidermal growth factor receptor), CD11a, CCR4 (chemokine C-C receptor 4), CD105, CD123, CD137, CD19, CD22, CD23, CD3, CD30, CD38, CD4, CD40, CD55SC-1, CD56, CD6, CD74, CD80, CS1 (cell surface glycoprotein 1), CTLA4 (cytotoxic T lymphocyte antigen 4, also known as CD152), DR5 (death receptor 5), Ep-CAM (epithelial cell adhesion molecule), folate receptor alpha, ganglioside GD2, ganglioside GD3, GPNMB, glycoprotein NMB, HGF / SF (hepatocyte growth factor / scatter factor), IGF-1 (insulin-like growth factor), IGF1-receptor (insulin-like growth factor-1 receptor), IL13 (interleukin-13), IL6 (interleukin-6), IL-6R (interleukin-6 receptor), immunodominant fungal antigens, heat shock protein 2. The metal fullerene-antibody complex of claim 1, which is selected from the group of antibodies or antibody fragments used in oncology targeting protein 90 (hsp90), integrin alpha5beta3, MHC (major histocompatibility complex) class II, MN-antigen (also known as G250 antigen), MUC1, PD-1 (programmed death 1), PIGF (placental growth factor), PDGFRa (platelet derived growth factor receptor alpha), prostate specific membrane antigen (PSMA), PTHrP (parathyroid hormone related protein), CD200 receptor, receptor activator of nuclear factor kappa B ligand (RANKL), sphingosine-1-phosphate (SIP), TGF-beta (transforming growth factor beta), TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) receptor 1, tumor necrosis factor receptor 2, vascular endothelial growth factor receptor 2 (VEGFR-2), CD33, CD20, CA125 (cancer antigen 125) or epidermal growth factor receptor.

4. 3. The metallofullerene-antibody complex of claim 2, wherein said antibody is selected from one or more of EGFR, HER2, HER3 and HER4 binding antibodies.

5. The metallofullerene-antibody complex of claim 2, wherein the antibody is selected from one or more of depatuxizumab, valisacumab, and trastuzumab.

6. 2. The metallofullerene-antibody complex of claim 1, wherein M is selected from one or more of holmium, lutetium, and scandium.

7. The metallofullerene-antibody complex of claim 1, wherein the proliferative disease is cancer.

8. 8. The metallofullerene-antibody complex of claim 7, wherein the type of cancer is selected from one or more of the group consisting of lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, colon / colorectal cancer, bladder cancer, thyroid cancer, breast cancer cells, liver cancer, ovarian cancer, endometrial cancer, cervical cancer cells, kidney cancer, brain cancer, or melanoma.

9. 2. The metallofullerene-antibody complex of claim 1, wherein said low level ionizing radiation ranges from about 0.02 mGy to about 60 Gy.

10. A pharmaceutical composition comprising the metallofullerene-antibody complex of claim 1 and a pharma- ceutically acceptable excipient, diluent or vehicle.

11. 11. The pharmaceutical composition of claim 10 in a form suitable for administration orally, intravenously, topically, by inhalation or via a suppository.

12. The metallofullerene-antibody conjugate of claim 1, further comprising an additional therapeutic agent.

13. A method for treating a cell proliferation associated disorder or disease, comprising administering to a patient of formula (I): AbC ｍ M ｎ @F ２ｖ (I) (Ab indicates cell binding agent; M represents a rare earth element in the periodic table of the elements selected from scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F to said Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer from about 20 to about 60. administering to the patient an effective amount of a metallofullerene-antibody complex of A method comprising administering a low level of ionizing radiation, wherein said metallofullerene-antibody complex is optionally therapeutically activated by said ionizing radiation.

14. The method of claim 13, wherein the Ab is a cell binding agent selected from the group comprising chimeric, CDR-grafted, humanized or recombinant human antibodies, more preferably, the Ab specifically binds to a tumor-associated antigen or cell surface receptor.

15. The Abs are: HER2, HER2, HER3, HER4 CD52, VEGF (vascular endothelial growth factor), EGFR (epidermal growth factor receptor), CD11a, CCR4 (chemokine C-C receptor 4), CD105, CD123, CD137, CD19, CD22, CD23, CD3, CD30, CD38, CD4, CD40, CD55SC-1, CD56, CD6, CD74, CD80, CS1 (cell surface glycoprotein 1), CTLA4 (cytotoxic T lymphocyte antigen 4, also known as CD152), DR5 (death receptor 5), Ep-CAM (epithelial cell adhesion molecule), folate receptor alpha, ganglioside GD2, ganglioside GD3, GPNMB, glycoprotein NMB, HGF / SF (hepatocyte growth factor / scatter factor), IGF-1 (insulin-like growth factor), IGF1-receptor (insulin-like growth factor-1 receptor), IL13 (interleukin-13), IL6 (interleukin-6), IL-6R (interleukin-6 receptor), immunodominant fungal antigens, fever syndrome 15. The method of claim 14, wherein the antibody or antibody fragment is selected from the group of antibodies or antibody fragments used in oncology targeting shock protein 90 (hsp90), integrin alpha5beta3, MHC (major histocompatibility complex) class II, MN-antigen (also known as G250 antigen), MUC1, PD-1 (programmed death 1), PIGF (placental growth factor), PDGFRa (platelet derived growth factor receptor alpha), prostate specific membrane antigen (PSMA), PTHrP (parathyroid hormone related protein), CD200 receptor, receptor activator of nuclear factor kappa B ligand (RANKL), sphingosine-1-phosphate (SIP), TGFbeta (transforming growth factor beta), TRAIL (tumor necrosis factor (TNF)-related apoptosis-inducing ligand) receptor 1, tumor necrosis factor receptor 2, vascular endothelial growth factor receptor 2 (VEGFR-2), CD33, CD20, CA125 (cancer antigen 125) or epidermal growth factor receptor.

16. 15. The method of claim 14, wherein the antibody is selected from one or more of EGFR, HER2, HER3 and HER4 binding antibodies.

17. 14. The method of claim 13, wherein the antibody is selected from one or more of depatuxizumab, valisacumab, and trastuzumab.

18. 14. The method of claim 13, wherein M is selected from one or more of holmium, lutetium, and scandium.

19. The method of claim 13, wherein the proliferative disease is cancer.

20. 20. The method of claim 19, wherein the type of cancer is selected from one or more of lung cancer, prostate cancer, head and neck cancer, pancreatic cancer, colon / colorectal cancer, bladder cancer, thyroid cancer, breast cancer cells, liver cancer, ovarian cancer, endometrial cancer, cervical cancer cells, kidney cancer, brain cancer, and melanoma.

21. 14. The method of claim 13, wherein the low level ionizing radiation ranges from about 0.02 mGy to about 60 Gy.

22. The method of claim 13, wherein the complex is contained in a pharma- ceutically acceptable composition comprising an excipient, diluent or vehicle.

23. 23. The method of claim 22, wherein the pharmaceutical composition is in a form suitable for administration orally, intravenously, topically, by inhalation, or via a suppository.

24. 14. The method of claim 13, further comprising administering one or more additional therapeutic agents.

25. The method of claim 13, wherein the metallofullerene-antibody complex is administered in an intravenous infusion dose.

26. The method of claim 13, wherein the conjugate is administered to the patient as a first line therapy.

27. 14. The method of claim 13, wherein the conjugate is administered to the patient as a second, third, fourth, fifth or sixth line treatment.

28. The method of claim 13, wherein the conjugate is administered to the patient after at least one treatment with an anti-cancer therapy.

29. 14. The method of claim 13, wherein the cell proliferation-related disorder or disease is resistant to at least one anti-cancer agent.

30. 14. The method of claim 13, wherein the method for treating a cell proliferation-associated disorder or disease inhibits metastasis in the patient.

31. 14. The method of claim 13, wherein the complex is administered to the patient and, once the complex has concentrated in the target cells, is followed by administration of a low level of ionizing radiation.

32. 32. The method of claim 31 , wherein the low level ionizing radiation is administered to the patient within 24 hours, preferably within 12 hours, more preferably within 6 hours of administration of the conjugate.

33. Formula (I): AbC ｍ M ｎ @F ２ｖ (I) (Ab indicates cell binding agent; M represents a rare earth element in the periodic table of the elements selected from scandium, yttrium, lanthanum, cerium, praseodymium, neodymium, promethium, samarium, europium, gadolinium, terbium, dysprosium, holmium, erbium, thulium, ytterbium, and lutetium, or combinations thereof; F represents a fullerene molecule having v carbon atoms; C represents a linker connecting fullerene F to said Ab; n represents an integer of 1 to 3; m represents an integer of 0 to 3; and v represents an integer from about 20 to about 60. US20100232633A1 - Encapsulated metallofullerene-antibody conjugates for use in the preparation of computed tomography contrast agents.

34. 1. A method for imaging a tumor in a patient, comprising: administering to said patient an effective amount of the endohedral metallofullerene-antibody complex of claim 33; and The method comprises exposing tissue of the patient to low levels of ionizing radiation.

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