MOF with β-emitters for radiation therapy

Abstract

The present invention relates to MOFs for use in radiotherapy. More specifically, the present invention provides particles comprising a MOF, optionally at least one targeting moiety, and at least one β-ray-emitting radionuclide; compositions comprising the particles; the particles or compositions for use as pharmaceuticals; the particles or compositions for use in methods for treating proliferative diseases; and kits comprising particles comprising a MOF, optionally a targeting moiety, and a β-ray-emitting radionuclide.

Description

[Technical Field]

[0001] The present invention relates to MOFs for use in radiotherapy. More particularly, the present invention provides particles comprising a MOF and at least one β-emitting radionuclide; compositions comprising the particles; the particles or compositions for use as a medicament; the particles or compositions for use in a method for treatment; and kits comprising particles comprising a MOF and optionally a targeting moiety, and a β-emitting radionuclide. [Background technology]

[0002] Common methods of cancer treatment include surgery, chemotherapy, and external beam radiation therapy, but there is a significant unmet medical need for new and more effective cancer treatment options.

[0003] One treatment option that promises high specificity and efficacy is radioimmunotherapy (RIT). RIT agents are based on a tissue-targeting moiety coupled to a carrier capable of binding and retaining a radioisotope. This tissue-targeting moiety, often an antibody, is used to deliver the radioisotope to the vicinity of unwanted cell types, such as cancer cells, resulting in the localized delivery of high-energy, cytotoxic radiation, killing the unwanted cells and sparing healthy surrounding tissue. Such specific cell killing may be essential for the treatment of cancerous diseases such as sarcomas and carcinomas, as well as hyperplastic, neoplastic, and chronic inflammatory diseases. Furthermore, systemic treatment with RIT is highly effective in patients with metastatic tumors, which would otherwise be difficult to achieve without adverse side effects.

[0004] Successful delivery of radiation to undesired cells depends on a stable interaction between the radioisotope and the carrier to prevent unintended side effects due to leakage of the radioisotope. However, currently used carriers (often small molecule ligands or chelators) do not meet these requirements for a wide range of potent radioisotopes. As a result, some of the most potent radioisotopes cannot be used in RIT. WO 2020 / 228558 describes radioactive particles produced by precipitation of radioactive materials, such as beta-emitters, within hydrophilic porous solid materials or carriers, particularly by adsorption onto fine particles of activated carbon and diatomaceous earth. One type of novel radioisotope carrier being investigated is crystalline porous coordination polymers (PCPs), also known as metal-organic frameworks (MOFs), as described in the applicant's WO 2023 / 282769. These are constructed by substituting inorganic and organic monomers for pores in a three-dimensional network. The interior pores of the MOF particles may have strong binding sites to which radioisotopes may be strongly bound, for example by chelation.

[0005] Solid tumors are known to be difficult to treat because they do not contain liquid regions. These non-vascularized tumors hinder immune cells and drug compounds from penetrating into the tissue, posing one of the major challenges in developing new therapies. Therefore, improved agents and methods for radiation therapy, particularly for the treatment of cancerous diseases, are needed. Summary of the Invention

[0006] The present inventors have discovered that certain metal-organic framework (MOF) structures containing specific free functional groups can adsorb and act as stable carriers of radionuclides in clinically relevant environments. It has been newly discovered that applicant's MOF structures containing specific free functional groups also function as stable carriers for beta-emitting radionuclides, and that these MOFs in particulate form are useful for the treatment of proliferative diseases. Furthermore, these MOFs, in particulate form, can be linked to targeting moieties, allowing for targeted delivery of the particles, and thus the beta-emitting radionuclides, in vivo.

[0007] Thus, the particles of the present invention containing beta-emitting radionuclides are promising drug candidates.

[0008] The particles of the invention comprising MOFs as carriers of beta-particle-emitting radionuclides can be used, for example, in the treatment of proliferative diseases, in particular non-vascular tumors.

[0009] In one aspect, the present invention relates to a particle, said particle comprising: a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, said functional group being selected from the group comprising carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs containing; at least one beta-emitting radionuclide; Including, wherein said at least one radionuclide is located in at least one of the pores.

[0010] The present invention also relates to targeted particles, said particles comprising at least one targeting moiety attached to the particle on the exterior surface of the particle.

[0011] In a further aspect, the present invention relates to a composition as further disclosed, said composition comprising at least one particle comprising at least one pharmaceutically acceptable carrier, diluent, and / or excipient, as defined.

[0012] In yet another aspect, the present invention relates to said particle or said composition for use as a medicament or for use in therapy, as further disclosed herein.

[0013] In a further aspect, The present invention relates to a kit, said kit comprising in a first container: a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOF containing including particles comprising; In the second container, Beta-emitting radionuclides Includes:

[0014] In one embodiment, the particle further comprises at least one targeting moiety linked to the particle on the exterior surface of the particle. [Brief explanation of the drawings]

[0015] [Figure 1] FIG. 1 is a schematic diagram illustrating one embodiment of the present invention. [Figure 2] Figure 2 is a schematic diagram of a network structure with an fcu topology. [Figure 3] FIG. 3 is a diagram showing the pore of fcuMOF. [Figure 4] FIG. 4 shows examples of coordination sites at the corners of a tetrahedral cage. [Figure 5]FIG. 5 shows examples of coordination sites at the corners of an octahedral pore. [Figure 6] FIG. 6 shows examples of coordination sites at the corners of an octahedral pore. [Figure 7] FIG. 7 shows examples of coordination sites at the corners of an octahedral cage. [Figure 8] FIG. 8 shows an example of a pore with multiple free functional groups extending into the pore. [Figure 9] FIG. 9 shows the MOF UiO-66. [Figure 10] FIG. 10 shows the MOF MIL-53. [Figure 11] FIG. 11 shows a MOF with a zeolitic imidazolate framework and the so-called sod topology. [Figure 12] FIG. 12 shows the BCA assay of antibody (IgG) conjugated UIO-66(COOH)2. [Figure 13] FIG. 13 shows that anti-CD37 conjugated MOF (NP1) binds to CD37-positive cells (Duadi cells). [Figure 14] FIG. 14 shows that anti-EpCAM or anti-HER-labeled UIO-66(COOH)2 binds to colon cancer cells (HTC116 and HT29) that express both antigens. [Figure 15] FIG. 15 shows that anti-HER2 binding UIO-66(COOH) 2 binds to HER2-positive cancer cells (JIMT1 cells). [Figure 16A] FIG. 16A shows the body weights of mice injected with radium-223 (Ra223)-loaded UIO-66(COOH)2 and that developed side effects at doses of 937 kBq per kg or higher. [Figure 16B] FIG. 16B shows the body weights of mice injected with radium-223 (Ra223)-loaded UIO-66(COOH)2 and that developed side effects at doses of 937 kBq per kg or higher. [Figure 17]FIG. 17 shows the biodistribution of free Lu177 and Lu177-labeled anti-EpCAM conjugated UiO-66-(COOH)2, demonstrating the in vivo retention of Lu177 in the MOF structure. [Figure 18] FIG. 18 shows the binding of Lu177-labeled anti-EpCAM conjugate UiO-66-(COOH)2 to cancer cells (HCT116) in vitro. DETAILED DESCRIPTION OF THE INVENTION

[0016] Unless otherwise defined, all terms, notations, and other scientific or technical terms used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this invention belongs. In some cases, terms having a commonly understood meaning are defined herein for clarity and / or ready reference, and the inclusion of such definitions herein should not necessarily be construed as representing a substantial departure from what is commonly understood in the art.

[0017] Metal-organic frameworks (MOFs), also known as multiporous coordination polymers (PCPs), are solid compounds, usually in powder form, belonging to the coordination polymer class. Chemically, MOFs comprise a repeating network of polytopic inorganic monomers, in the form of metal ions or clusters, and organic monomers, also called bridging ligands or linkers. The inorganic and organic monomers are linked by coordinate covalent or ionic bonds between Lewis acidic metal cations and Lewis basic organic functional groups (e.g., carboxylates, amines, imines, etc.). The structure of a MOF can be well tunable through the selection of organic and inorganic monomers. The organic monomers of a MOF can all be identical, or the MOF can be composed of two or more different organic monomers with nearly identical framework coordination functional group configurations. The latter are called "mixed-linker MOFs." Similarly, the inorganic monomers of a MOF can also be identical, or they can be different, in which case they are called "mixed-metal MOFs."

[0018] MOFs can be crystalline, amorphous, or have a deformable structure. MOFs made from structurally rigid monomers tend to form repeating three-dimensional networks characterized by permanent porosity, i.e., they are crystalline. Such crystalline MOFs are characterized by numerous pores. As used herein, the term "pore" refers to any type of opening, cavity, channel, or hole within the MOF network. MOFs can contain pores of various shapes in different proportions based on the MOF's crystalline structure. However, the pores of a particular MOF are generally very similar to each other in terms of size and chemical environment.

[0019] The organic monomers of MOFs may contain functional groups that do not directly interact with the inorganic monomers to form the MOF network. In this disclosure, such functional groups are referred to as "free" functional groups. When such functional groups are present in a crystalline MOF, they can be introduced into the pores of the MOF to form sites for adsorption, coordination, and / or chelation of guest ions or molecules. The exact nature of such sites depends on the network topology of the MOF and the type and number of functional groups introduced into the pores. By selecting appropriate MOFs in terms of free functional groups, specific chelating sites, such as within the pores, can be constructed for strong and selective adsorption, coordination, and / or chelation of specific compounds. As used herein, the terms "chelating" and "chelation" refer to the coordination, complexation, and / or binding of a metal, e.g., a radionuclide cation, by at least two ligands. The term "ligand" refers to a moiety, eg, a molecule, a portion of a molecule, a functional group, etc., that is capable of coordinating, complexing and / or bonding to a metal.

[0020] Various functional groups can be used to modify the shape and charge balance of the chelating moieties. Because MOFs are constructed from coordinate bonds between organic and inorganic monomers, adding functional groups that can act as coordinating groups may produce undesired and / or unexpected products during MOF preparation. Therefore, the introduction of free functional groups into MOFs requires careful design.

[0021] The paper “Sr. 2+ "Carboxyl-functionalized UIO-66(COOH)2 for selective adsorption of radioactive contaminants" (Gao, Y. et al, Molecules 2022, 2022, 27(4), 1208) demonstrates that the functionalized MOF UIO-66(COOH)2 can be used for selective adsorption of radioactive contaminants. 90reported that UIO-66(COOH)2 can be used for the selective removal of Sr. This study shows that the framework of UIO-66(COOH)2 is characterized by non-bonded carboxyl groups, high stability, and porosity, which enable selective and effective adsorption of Sr ions. Gao et al. reported that UIO-66(COOH)2 not only has high adsorption capacity, but also has a similar NA radius. + YaK + in the presence of Sr ions 2+ This MOF also enables selectivity for radioactive Sr in contaminated water. 2+ They conclude that UIO-66(COOH)2 may be useful in the removal of strontium. Gao et al. make no mention of the potential usefulness of UIO-66(COOH)2 in medical therapy or of other radionuclides suitable for use in radiotherapy in medical therapy. Sr-90 (used by Gao et al.) has a half-life of 29 years and is therefore not relevant for use as a therapeutic radioisotope.

[0022] In the present invention, particles of a specific type of MOF are used as carriers for β-ray-emitting radionuclides suitable for use in therapy. The MOFs used in the present invention are crystalline MOFs containing free functional groups that can be metal-coordinating and that extend into the pores of the MOF in such a manner that the free functional groups can coordinate and / or bind to β-ray-emitting radionuclides, such as the cations of the radionuclides. Thus, each pore of these MOFs has the ability to function as a chelating macroligand for the radionuclide. This MOF particle is a class of carriers that can contain potent β-ray-emitting radioisotopes with little or no leakage, thereby improving the efficiency of RIT and providing new options for the treatment of proliferative diseases such as cancer.

[0023] Solid tumors are known to be difficult to treat because they do not contain liquid regions. These non-vascularized tumors hinder immune cells and pharmaceutical compounds from penetrating into the tissue, posing one of the major challenges in developing new therapeutics. Targeted therapy using β-emitters has the potential to bind to the non-vascular tumor surface and irradiate multiple layers within the tumor tissue that would otherwise be hidden. Applicant has discovered that the MOF particles of the present invention are suitable for delivering β-emitters to such tumors and tissues, enabling their treatment.

[0024] Thus, in one aspect, the present invention relates to a particle comprising a MOF, wherein the MOF comprises a repeating three-dimensional network of inorganic and organic monomers forming pores, the pores being designed for chelating a radionuclide, and the particle comprises a β-emitting radionuclide located within at least one of the pores. Optionally, the particle comprises at least one targeting moiety. Thus, in one embodiment, the particle comprises at least one targeting moiety attached to the particle on the outer surface of the particle.

[0025] In some embodiments, the present invention relates to a particle, the particle comprising: a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs containing; Optionally, at least one targeting moiety linked to the particle on the outer surface of the particle; and at least one beta-emitting radionuclide; Including, wherein said at least one radionuclide is located in at least one of the pores.

[0026] In these embodiments, at least one targeting moiety may be linked to the particle on the exterior surface of the particle.

[0027] FIG. 1 shows a schematic representation of a non-limiting embodiment of a particle 1 of the present invention comprising a MOF 2 that forms a pore 3 and has a functional group 4 extending into the pore 3, where the particle 1 comprises a targeting moiety 5, shown here as an antibody, and a radionuclide 6 is located within the pore 3.

[0028] The MOFs of the present invention comprise repeating three-dimensional structures of inorganic and organic monomers, i.e., they are crystalline. The organization of the inorganic and organic monomers in a repeating three-dimensional network results in the formation of pores. That is, the rigidity and set geometry of the inorganic and organic monomers result in a set spatial separation between specific monomers, resulting in the formation of pores. MOFs with free functional groups can be understood to have somewhat dynamic pore sizes due to the freedom of orientation of the free functional groups. Pore size can be measured, for example, by the "inclusion sphere," i.e., the largest sphere that can be placed within the pore without contacting the van der Waals surface of the MOF. The size of the pore opening, also referred to as the "window size," can be measured by the "diffusion sphere," i.e., the smallest sphere that can move within the structure. MOFs for use in the present invention advantageously have inclusion sphere pore sizes of 0.3 to 3 nm, preferably 0.5 to 1.5 nm. Furthermore, the MOFs of the present invention have pore windows of a certain size, preferably diffusion spheres with a diameter of at least 3 Å, that allow the adsorption of radionuclides. The pore size can be controlled by the size, structure, and bonding of the inorganic and organic monomers. The pores can be of any shape, including, but not limited to, tetrahedrons, octahedrons, and hexahedrons.

[0029] Of the approximately 100,000 or more MOF structures reported, only a limited number can be utilized in the present invention. Useful MOFs are essentially limited by their pore topology and their ability to accommodate functional groups on linkers that point into the pore instead of being attached to inorganic monomers, or by post-synthetic incorporation onto either the linker or the inorganic cluster. The pore topology of the MOF must allow diffusion of radioactive ions from the exterior into the pore, and the pore must be large enough to accommodate functional groups and adsorbed radioactive cations. Examples of MOF structures that fit this description include, for example, the list of MOFs provided herein. Suitable pore sizes and pore window sizes are described above.

[0030] Because radioactive cations are adsorbed into negatively charged pockets in MOFs, the aforementioned functional groups should have a negative or partially negative polarity in an aqueous environment at physiological pH. A list of such functional groups is provided herein. In some cases, such functional groups are directly accessible following MOF synthesis, such as UiO-66(COOH)2 and UiO-66-NH2, with the carboxylate and amine groups pointing into the pore, respectively. In some cases, the functional groups are accessible through post-synthetic modification; for example, ZIF-90, which contains an aldehyde group pointing into the appropriate pore, can be converted to ZIF-8-COOH by reaction with hydrogen peroxide. In other cases, the functional groups are incorporated onto inorganic monomers, such as the introduction of a free imidazolate group into MOF-808 by reaction with histidine.

[0031] MOFs based on Zr(IV), Hf(IV), Ce(IV), Fe(III), Al(III), Ti(IV), Cr(III) with carboxylate organic monomers and Zn(II) imidazolate are known to be stable in aqueous media and therefore may be particularly useful in the present invention.

[0032] An example of a preferred MOF is terephthalate (C6H4(COO)2 2- ) Cationic hexanionic zirconium cluster Zr6O4(OH)4 as organic and inorganic monomers 12+

[0023] Other preferred examples of MOFs are the zirconium-based UiO-66, in which each carboxylic acid group of the terephthalate is coordinated to two cations of an inorganic monomer.

[0024] Other preferred examples of MOFs are the corresponding hafnium and cerium derivatives of UiO-66, Hf-UiO-66 and Ce-UiO-66 (Hf6O4(OH)4, respectively). 12+ and Ce6O4(OH)4 12+ Another example of a zirconium-based MOF is MOF-808, C24H16O32Zr6, which contains a benzene-1,3,5-tricarboxylic acid linker. UiO-66, Hf-UiO-66, Ce-UiO-66, and other MOFs with the same topology (network shape), known as the fcu network topology, contain tetrahedral and octahedral pores, with a dynamic ratio of 2:1 between the former and the latter. Each tetrahedral pore contains four corners where three organic linkers "meet," i.e., extend toward the same spatial volume, and each octahedral pore contains six corners where four organic linkers "meet." In their most stable conformation, the free functional groups are oriented toward the corners of the octahedral pore, but dynamic rotation around the linker axis and steric repulsion between adjacent carboxylate groups can often direct the free functional groups into the tetrahedral pore as well.

[0033] Figure 2 shows a schematic diagram of a network structure with the fcu topology, where inorganic monomers are shown as spheres with a cuboctahedral coordination geometry and organic monomers as bridging rods. Figure 3 shows the octahedral (O) and tetrahedral (T) pores of the fcuMOF. The large transparent spheres indicate the size of each pore.

[0034] In one embodiment, the inorganic monomer is metal-based in the M(IV) oxidation state. M(IV)-based MOFs where M = Zr and / or Hf, i.e., MOFs in which the inorganic monomer is based on the metal Zr and / or Hf in the +4 (M(IV)) oxidation state, and which have carboxylate-based organic monomers, such as UiO-66, Hf-UiO-66, and their derivatives, are considered particularly suitable for in vivo applications for several reasons: (1) the monomers may have advantageously low toxicity in vivo; (2) the MOFs may exhibit excellent stability in aqueous solutions, such as human serum, due to the stability of the M(IV)-carboxylate bond against hydrolysis, as shown in Examples 2 and 3; and (3) the M(IV) oxocluster inorganic monomers may have high connectivity, i.e., a large number of organic monomers may be linked together such that the free functional groups of several organic monomers are in close spatial proximity (which may allow the formation of highly coordinated chelating sites adjacent to the cluster). As a result, the MOF particles of the present invention are particularly stable and efficient carriers for radionuclides, which may have very high adsorption capacities, allowing for specific treatment with negligible leakage to other organs.

[0035] Mixed-metal and / or mixed-linker derivatives of UiO-66 are also expected to exhibit advantageous properties similar to those of UiO-66. Related mixed-metal derivatives include derivatives containing two or more metals from the group including or consisting of Zr(IV) and Hf(IV) in inorganic monomers. Related mixed-linker derivatives include derivatives containing monomers known to those skilled in the art to have coordination geometries similar to terephthalates, such as monomers selected from the group including or consisting of aminoterephthalate, hydroxyterephthalate, mellitate, pyromellitate, sulfoterephthalate, muconate, and combinations thereof. Suitable mixed-linker MOFs can be obtained by multiple linker synthesis or by linker exchange, both of which are well known to those skilled in the art.

[0036] Additionally, MOFs with inorganic monomers based on Fe(III), Al(III), Ti(IV), and / or Cr(III) may also be particularly useful in the present invention. The organic monomers may be selected from the group including or consisting of, for example, terephthalate, aminoterephthalate, hydroxyterephthalate, mellitate, pyromellitate, sulfoterephthalate, muconate, and combinations thereof. While the MOFs may not form the same topology as UiO-66, they may form other interesting topologies that may function in the same manner.

[0037] The MOFs of the present invention comprise at least one free functional group extending into the pores. Preferably, the MOFs comprise a repeating three-dimensional network of inorganic and organic monomers that form pores, with at least one free functional group extending into each pore (i.e., each pore has at least one free functional group extending therein). In some embodiments, only a certain amount of the pores, such as 50% of the pores, e.g., 80% of the pores, e.g., 95% of the pores, e.g., 100% of the pores, have at least one free functional group extending therein.

[0038] As used herein, the term "extending into a pore" means that the functional group is directed, oriented, and / or present in one of the pores formed by the repeating three-dimensional network of inorganic and organic monomers. Thus, the free functional group is available within the pore to coordinate, bind, and / or react with compounds, ions, and the like. In particular, the functional group is available to coordinate, complex, and / or bind to a radionuclide, e.g., a radionuclide cation.

[0039] At least one free functional group is attached to an inorganic monomer and / or an organic monomer. When at least one free functional group is attached to an organic monomer, it is preferably covalently attached to the organic monomer. In some embodiments, each organic monomer of the MOF has at least one free functional group attached thereto. In other embodiments, a predetermined amount of organic monomers each have at least one free functional group attached thereto. In one embodiment, at least one free functional group is attached to an organic monomer. Furthermore, preferably, the MOF comprises at least one free functional group that results in a net negative charge in the pore. In some embodiments, each inorganic monomer of the MOF has at least one free functional group attached thereto. In other embodiments, a predetermined amount of inorganic monomers each have at least one free functional group attached thereto.

[0040] The free functional groups are preferably Lewis basic and are selected from the group including or consisting of carboxylic acids, carboxylates, hydroxyls, sulfonic acids, sulfonates, sulfhydryls, primary amines, secondary amines, and combinations thereof. Secondary amines are selected from the group including or consisting of amines of the general structural formula -NHR, where R is selected from the group including or consisting of C1-C8 alkyls, alkenyls, and alkynyls having at least one Lewis basic functional group (e.g., carboxylic acids, carboxylates, hydroxyls, sulfhydryls, sulfonic acids, sulfonates, primary amines, secondary amines, and combinations thereof), such as C1-C4 linear and branched alkyls, alkenyls, and alkynyls having at least one Lewis basic functional group (e.g., carboxylic acids, carboxylates, hydroxyls, sulfonic acids, sulfonates, sulfhydryls, primary amines, secondary amines, and combinations thereof). Such functionality can be obtained, for example, by performing a peptide condensation between an N-protected natural amino acid and the free primary amino group of the organic monomer of the MOF.

[0041] The free functional groups may be advantageously selected based on the choice of radionuclide. For example, if the radionuclide is known to those skilled in the art to be acidophilic, or "hard" according to the HSAB theory (hard and soft (Lewis) acids and bases), the pore may advantageously comprise at least one oxygen-containing functional group, i.e., carboxylic acid, carboxylate, hydroxyl, sulfhydryl, sulfonic acid and / or sulfonate. Correspondingly, if the radionuclide is "soft" according to the HSAB theory, the pore may advantageously comprise at least one amine group.

[0042] Thus, in some embodiments, the MOF comprises at least one free functional group selected from the group comprising carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof. In other embodiments, the MOF comprises at least one free functional group selected from the group comprising or consisting of carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfhydryl, sulfonate, and combinations thereof. In still other embodiments, the MOF comprises at least one free functional group selected from the group comprising or consisting of primary amine and secondary amine, and combinations thereof. In some embodiments, the MOF comprises at least one free functional group selected from the group comprising carboxylic acid, carboxylate, primary amine, and secondary amine.

[0043] Preferably, the MOF comprises 1 to 4 free functional groups, for example 1 to 2 functional groups, for example at least 3, for example at least 4 free functional groups. In some embodiments, the MOF comprises at least 2 free functional groups, and the free functional groups are the same, or at least 2, for example at least 3, for example all, of the free functional groups are different from each other. In some embodiments, the MOF comprises at least 3 free functional groups, and the free functional groups are the same, or at least 2, for example at least 3, for example all, of the free functional groups are different from each other. In some embodiments, the MOF comprises at least 4 free functional groups, and the free functional groups are the same, or at least 2, for example at least 3, for example all, of the free functional groups are different from each other.

[0044] When the MOF comprises at least two free functional groups as disclosed above, the structure of the MOF can be selected to allow some or all of the at least two free functional groups to point toward a common spatial volume, such as a common point or volume within a pore. When the MOF comprises at least two free functional groups as disclosed above, the structure of the MOF can be selected to allow some or all of the at least two free functional groups to orient in a manner that allows for favorable interactions with the radionuclide cation in the presence of the radionuclide cation, such as by aligning with the outer orbital of the radionuclide cation. In some embodiments, each free functional group has a distance of 4 to 15 Å, e.g., 4 to 12 Å, e.g., 5 to 10 Å, from at least one, e.g., all, of the other free functional groups extending into the same pore.

[0045] Furthermore, the selection and placement of the at least one free functional group can be used to alter the geometry of the coordination, e.g., binding, e.g., chelation, etc., site within the pore. Such sites can be formed by the presence of one or more free functional groups, but can also result from the size and / or shape of the pore. In some embodiments, such sites are defined as including all free functional groups extending into the same pore. In some embodiments, such sites are defined as including free functional groups extending into the same pore and having a maximum distance of no more than 15 Å, e.g., 10 Å, from at least one other free functional group within the same pore. In some embodiments, such sites are present at the corners of the pore. This placement may entail that the site is only accessible from one direction, thus limiting competitive adsorption. Furthermore, if such sites are present at the corners of the pore or otherwise within the interior of the pore opposite the pore opening, other compounds, e.g., other cations, e.g., competing cations, may be present within the pore and sterically block the radionuclide from leaving the pore. Such corner sites can, in principle, be constructed in all MOFs capable of containing free functional groups, including Cr-MIL-100, Fe-MIL-100, and Al-MIL-100, Cr-MIL-53, Fe-MIL-53, and Al-MIL-53, Zr-MIL-140, Hf-MIL-140, Zr-MOF-711, Hf-MOF-711, ZIF-8, and Ti-MIL-125.

[0046] Examples of coordination, binding, and / or chelation sites are shown in Figures 4-7. Figure 4 shows an example of a coordination, binding, and / or chelation site at the corner of a tetrahedral cage with three free carboxylate groups pointing into the pore. Figure 5 shows an example of such a site at the corner of an octahedral pore with four free carboxylate groups pointing into the pore. Figure 6 shows such a site at the corner of an octahedral pore with two free carboxylate groups and two free amide groups pointing into the pore. The latter can be obtained, for example, by post-synthetic peptide condensation on free primary amino groups. Post-synthetic modifications are discussed below. Figure 7 shows such a site at the corner of an octahedral cage with two free carboxylate groups and two free amide groups pointing into the pore, one of which carries a DOTA chelator. Figure 8 shows a pore with multiple free functional groups extending into the pore.

[0047] Furthermore, at least one functional group directed into the pore can be used to alter the charge balance of the pore and / or the charge balance of sites for coordination, e.g., binding, e.g., chelation, etc., within the pore. Advantageously, the MOFs of the present invention contain a free functional group, or combination of functional groups, that results in a net negative charge within the pore. This net negative charge may promote strong, selective adsorption of cations.

[0048] Introduction of substituents onto the phenyl ring of the terephthalic acid monomer of UiO-66 and UiO-66-type MOFs provides easy access to crystalline MOFs containing at least one free functional group extending into the pore, where the at least one free functional group is selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof. For example, UiO-66-COOH, UiO-66(COOH)2, and UiO-66-NH2 are all commercially available. UiO-66-COOH, UIO-66(COOH)2, Hf-UiO-66-COOH, and Hf-UIO-66(COOH)2 represent preferred MOFs with 6–12 free carboxylic acid functional groups extending into their pores, of which 3 or 4 free functional groups form coordination sites within the pore and thus can chelate radionuclide cations in an efficient and / or stable manner.

[0049] There are two main approaches to so-called pore engineering, i.e., the introduction of free functional groups extending into the pores. These include the synthesis of MOFs using "preintegrated organic monomers," which contain two different coordination sites for framework construction and other applications, and "postsynthetic modification," in which additional functional groups are sequentially attached to preassembled MOFs. Free functional groups in MOFs can be modified by chemical reactions commonly referred to as "postsynthetic modification." In principle, any chemical reaction that can be performed by standard wet chemistry techniques can be performed on the same functional group in a MOF. For example, amino acids can be reacted with free primary amine groups in a MOF by peptide condensation reactions. Using this method, molecules with specific properties can be grafted onto MOFs, such as into their pores. Therefore, MOFs with specific structures and specific free functional groups can be obtained by both designing and synthesizing new MOFs and modifying existing ones. Thus, various derivatives of MOFs having the UiO-66 structure, as well as MOFs having other structures and containing various free functional groups, are readily available. In some embodiments, at least one radionuclide chelator known to those skilled in the art, such as EDTA, e.g., DOTA, is grafted onto the group extending into the pore, such that at least one free functional group is a known chelator.

[0050] Furthermore, most MOFs can tolerate a percentage of linker-deficient defects, i.e., sites where an organic linker is missing from the structure, leaving a coordinatively unsaturated site on the adjacent inorganic monomer. These unsaturated sites can be modified with coordinating molecules, such as molecules with at least one functional group that can coordinate inorganic monomers and contribute to the functional properties of the MOF, such as amino acids, that have a free functional group for extending into the pore.

[0051] Preferably, the MOFs of the present invention consist of a repeating three-dimensional network of inorganic and organic monomers that form pores, and at least one free carboxylic acid or carboxylate functional group extending into each pore.

[0052] 9, 10 and 11 show schematic diagrams of preferred MOFs.

[0053] Figures 9 and 10 show partial crystal structures of M(IV)- and M(III)-based terephthalic acid MOFs. Figure 9 shows UiO-66 (organic monomers are shown as rods with C / O atoms at the corners, and inorganic monomers are shown as polyhedra). Figure 10 shows MIL-53, where the available pore space is shown as large spheres in addition to lines and polyhedra.

[0054] Preferred MOFs shown in Figures 9 and 10 may have an M(IV) inorganic monomer. Suitable organic monomers are shown below (structures I, II, III, IV, V, and VI): [ka]

[0055] Structure I represents terephthalic acid / terephthalate organic monomers, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds. 11 , R 12 , R 13 , and R 14 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0056] Structure II represents the biphenyl-4,4'-dicarboxylic acid organic monomer, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds.21 , R 22 , R 23 , R 24 , R 25 , R 26 , R 27 and R 28 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0057] Structure III represents trimesic acid (1,3,5-benzenetricarboxylic acid) organic monomer, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds. 31 , R 32 and R 33 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0058] Structure IV represents an adipic acid organic monomer, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds. 41 , R 42 , R 43 and R 44 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0059] Structure V represents a 1,4-cyclohexyldicarboxylic acid organic monomer, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds. 61 , R 62 , R63 , R 64 , R 65 , R 66 , R 67 , R 68 , R 69 and R 70 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0060] Structure VI represents a naphthyl organic monomer, which can form MOFs with M(III) and M(IV)-based inorganic monomers via metal-carboxylate bonds. 71 , R 72 , R 73 , R 74 , R 75 and R 76 may each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each of the R groups may be hydrogen or a free functional group.

[0061] In each of structures I-VI, the secondary amine can be selected from the group including or consisting of amines of the general structural formula -NHR, where R is selected from the group including or consisting of C1-C8 alkyl, alkenyl, and alkynyl having at least one Lewis basic functional group (e.g., carboxylic acid, carboxylate, hydroxyl, sulfhydryl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof), such as C1-C4 straight-chain and branched alkyl, alkenyl, and alkynyl having at least one Lewis basic functional group (e.g., carboxylic acid, carboxylate, hydroxyl, sulfhydryl, sulfonic acid, sulfhydryl, primary amine, secondary amine, and combinations thereof).

[0062] Figure 11 shows a MOF with a zeolitic imidazolate framework and the so-called sod topology, where the organic monomers are shown as balls and sticks representing atoms and bonds, and the Zn atoms are shown as tetrahedra.

[0063] A preferred MOF shown in Figure 11 may have an M(II) inorganic monomer. A suitable organic monomer is shown below (Structure VII): [ka]

[0064] Structure VII represents an imidazole organic monomer, which can form MOFs (also known as ZIFs) with M(II)-based inorganic monomers via metal-imide bonds. 51 , R 52 and R 53 can each independently be selected from the group including or consisting of hydrogen, carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, and secondary amine, i.e., each R group can be hydrogen or a free functional group. Secondary amines can be selected from the group including or consisting of amines of the general structural formula -NHR, where R is selected from the group including or consisting of C1-C8 alkyl, alkenyl, and alkynyl having at least one Lewis basic functional group (e.g., carboxylic acid, carboxylate, hydroxyl, sulfhydryl, sulfonic acid, sulfonate, primary amine, secondary amine, and combinations thereof), such as C1-C4 linear and branched alkyl, alkenyl, and alkynyl having at least one Lewis basic functional group (e.g., carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof).

[0065] In the inventions disclosed herein, MOFs exist in the form of particles, such as microparticles or nanoparticles. Preferably, the particles are nanoparticles. As used herein, the term "nanoparticle" refers to any particle having a diameter less than 1000 nm, e.g., 1-1000 nm. Similarly, the term "nanoparticle" refers to a plurality of particles having an average diameter of about 1-1000 nm. References to the "size" of nanoparticles refer to the length of the nanoparticle's largest linear dimension. For example, the size of a perfectly spherical nanoparticle is its diameter. Size may refer to the hydrodynamic radius of the particle, as characterized by, for example, dynamic light scattering. In some embodiments, the nanoparticles of the present invention have a particle diameter of 1-200 nm. Correspondingly, the term "microparticle" refers to a particle having a diameter less than 1000 μm, e.g., 1-1000 μm.

[0066] Microparticles and nanoparticles have also been used in radiation therapy. A key factor when using such particles is their stability, both in the sense that the particle itself should be completely stable or slowly degrade to reduce the risk of system toxicity, and in the sense that the radionuclide should be tightly bound to the particle to avoid leakage. The MOFs of the present invention are highly stable, as discovered by the inventors and shown in Examples 2 and 3. Furthermore, the presence of free functional groups, along with the confinement effect of the pores themselves, ensures strong chelation of the radionuclide to the particle. As shown in Example 7, the β-emitter lutetium-177 (Lu-177) chelated to UIO-66(COOH)2 showed high radionuclide adsorption to MOF particles and retention of Lu-177 in MOFs in serum. The choice of free functional groups can result in high levels of selectivity, e.g., preferential chelation of cations over anions and / or hard ions over soft ions. Thus, MOFs suitable for carrying useful radioisotopes, typically alpha emitters, for use in therapy as previously discovered by applicants have now been found to also be useful as carriers of beta-emitting radionuclides. Thus, the present invention provides particularly stable carriers for beta-emitting radionuclides.

[0067] Throughout the MOF particles of the present invention, there are 10^3 to 10^5 pores that have the potential to capture radionuclide cations in a portion of them, i.e., the portion accessible from the outside of the particle. If a radionuclide cation is released from one pore, it can be captured by an adjacent pore, preventing leakage from the particle. This ability to "readsorb" desorbed radionuclide cations indicates that the MOF particles of the present invention can more efficiently retain radionuclide cations compared to conventional chelators. Furthermore, the number of cations per particle can be adjusted, thereby allowing the level of radioactivity of the particle to be tuned.

[0068] Due to the presence of pores with suitable diffusion spheres and free functional groups extending into the pores, when the particles according to the invention are exposed to a solution containing radionuclide cations, these cations are easily adsorbed and chelated within the pores. A further advantage of using MOF particles according to the invention is that any competing species, such as other cations present in the solution, can occupy the missing pores rather than displacing the radionuclide cations.

[0069] The MOFs and particles thereof of the present invention can be obtained by any method known to those skilled in the art, such as by synthesis in a commercially available manner, for example using any synthesis protocol available to those skilled in the art.

[0070] In some embodiments, the particle of the present invention comprises at least one target site connected to the particle on the outer surface of the particle.The term "external surface" used herein in relation to particle refers to its outer surface that corresponds to the surface of the pore in the particle.When the pore penetrates the particle and is visible in the particle, the outer surface is defined to include all surfaces of the outermost surface of the particle, but does not include the surface that defines the pore.

[0071] As used herein, the term "targeting moiety" refers to a moiety, such as a molecule, e.g., a portion of a molecule, that is "tissue-targeting," i.e., that serves to preferentially localize itself—and any moiety, such as a particle to which it is linked—to at least one tissue site where it is present, e.g., for the delivery of a beta-emitting radioactive particle. The term "targeting moiety" can also refer to a functional group that serves to target or direct a particle to a specific location, cell type, diseased tissue, or association. A targeting moiety can be, for example, a moiety known to those skilled in the art to bind or complex to a biomarker, such as a cell surface marker, e.g., a receptor, transport protein, or cell adhesion molecule, present on diseased cells or cells in the vicinity of such cells. Such cell surface markers include, but are not limited to, proteins that are more highly expressed on diseased cell surfaces than on healthy cell surfaces, or proteins that are more highly expressed on cell surfaces during cell growth or replication than during quiescence. Moieties present in the vicinity of or associated with target cells or tissues can also be utilized in therapeutic targeting according to any embodiment of the present invention. For example, components present in or released from the matrix surrounding the target cells or tissue can be used for targeting if their presence, morphology, or concentration allows the region to be distinguished from healthy tissue.

[0072] Non-limiting examples of target sites include: carbohydrates; monosaccharides, such as glucose, mannose, galactose; urea derivatives; lipids; streptavidin; albumin; biotin; small molecule targeting moieties, such as steroid and non-steroid hormones; Aptamers, i.e., single-stranded oligonucleotides that recognize specific binding domains of receptors; Polymers such as biopolymers, e.g., antibodies; peptides, e.g., biomimetic peptides, e.g., phage-displayed peptides; proteins; nucleic acids; and cells, such as naturally occurring cells, e.g., genetically engineered cells Examples include:

[0073] Preferred targeting moieties include antibodies, antibody fragments, antibody constructs, constructs of antibody fragments, minibodies, nanobodies, intrabodies, unibodies, affibodies, and diabodies. As used herein, the term "antibody" refers to immunoglobulins, derivatives thereof that retain specific binding ability, and proteins having binding domains that are homologous or largely homologous to immunoglobulin binding domains. These proteins may be naturally occurring or partially or wholly synthetically produced. The term "antibody" encompasses, for example, monoclonal antibodies, polyclonal antibodies, recombinant antibodies, humanized antibodies, and / or chimeric antibodies. An antibody may be a member of any immunoglobulin class, including any of the human classes: IgG, IgM, IgA, IgD, and IgE. Examples of useful antibodies include conventional full-length antibodies (e.g., IgG) and camelid heavy chain antibodies (VHH).

[0074] As used herein, the term "antibody fragment" refers to any derivative of an antibody that is less than full-length. In exemplary embodiments, an antibody fragment retains at least a substantial portion of the full-length antibody's specific binding ability. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, scFv, Fv, dsFv diabody, and Fd fragments.

[0075] Cell-based targeting is in its early stages of development but may offer advantages such as high specificity and versatility. Therapeutic agents can be bound to the cell surface or encapsulated within the cell. Many cell types, including red blood cells, white blood cells, stem cells, platelets, dendritic cells, and even bacteria, have been exploited for tissue-specific delivery of small molecules, macromolecules, and even nanoparticles. When cells are used as targeting moieties, the treatment of the present invention can be considered cell therapy. The use of the particles of the present invention containing cells, such as CAR / TCR T cells, CAR / TCR NK cells, and NK-92, as targeting moieties in allogeneic or autologous cell therapy can provide both potent tumor-killing effects and immunomodulatory effects that prevent adverse side effects of cell therapy (e.g., cytokine release syndrome, graft-versus-host disease).

[0076] In some embodiments, the targeting moiety is one of two or more components that collectively have the effect of targeting the particles of the invention to a desired tissue. This is the case, for example, when one component binds to a specific tissue, tumor, or cell type (tissue-binding agent), and a second or additional component, the targeting moiety, binds to the tissue-binding agent. Suitable specific binding pairs for conferring mutual affinity between the tissue-binding agent and the targeting moiety are known in the art (e.g., biotin and avidin or streptavidin, etc.).

[0077] Targeting moieties can be obtained by any method known to those of skill in the art, e.g., commercially available, synthesized using any synthesis protocol available to those of skill in the art, e.g., produced enzymatically, synthetically and / or chemically.

[0078] In some embodiments, the particle contains only one targeting moiety. In other embodiments, the particle contains two or more targeting moieties, for example, five, for example, ten targeting moieties. The number of targeting moieties can be selected based on the size of the particle. The targeting moieties can be the same or different. The mass ratio of targeting moieties to particles can depend on the molecular weight of the targeting moieties and the diameter of the particles.

[0079] The use of two or more different targeting moieties has the advantage that more than one antigen can be targeted, resulting in improved binding to tissues or cells expressing the antigen profile associated with the target (e.g., M2 tumor-associated macrophages expressing CD163 and CD206). Another advantage would be the ability to target not only tumor-specific antigens but also blood vessels that accumulate at tumor sites (e.g., VCAM1).

[0080] At least one targeting moiety can be directly linked to the particle, for example, via a covalent bond, or at least one targeting moiety can be linked to the particle via a linking group. The targeting moiety can also be linked to the particle covalently conjugated with streptavidin, which strongly binds to biotin / biotin derivatives. Streptavidin can not only function as a linker but also as a binding site for the biotinylated moiety. Thus, streptavidin-conjugated particles can be bound to one or more biotinylated targeting moieties. Methods for linking targeting moieties to surfaces are well known in the art; for example, standard organic and / or inorganic chemistry, such as carbodiimide coupling, can be used. Suitable linking groups can be easily determined by those skilled in the art; for example, a range of linking groups is known from the field of antibody-drug conjugates. Non-limiting examples of linking groups include poly(ethylene glycol) (PEG), 2-(maleimidomethyl)-1,3-dioxane (MD), and maleimidocaproylsuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC). The exterior surface of MOF particles may be composed of the same organic functional groups as the interior of the pores and may undergo the same reactions. However, by using reagents that are sterically prohibited from entering the MOF pores, size discrimination can be used to perform reactions on the exterior surface rather than within the pores, thereby allowing for selective attachment of target moieties to the exterior surface of the particle.

[0081] The at least one targeting moiety may be a targeting moiety for targeting cells suffering from a proliferative disease, such as tumor cells, cancer cells, cells suffering from a hyperplastic disease, or cells suffering from a neoplastic disease. In a preferred embodiment, the targeting moiety is a targeting moiety for targeting cancer cells. In one embodiment, the targeting moiety targets cells of a primary tumor, such as a sarcoma. As used herein, the terms "cancer cell" and "tumor cell" refer to cells that divide at an abnormally increased rate. Cancer cells include, but are not limited to, carcinomas such as squamous cell carcinoma, non-small cell carcinoma (e.g., non-small cell lung cancer), small cell carcinoma (e.g., small cell lung cancer), basal cell carcinoma, sweat gland carcinoma, sebaceous gland carcinoma, adenocarcinoma, papillary carcinoma, papillary adenocarcinoma, bladder adenocarcinoma, medullary carcinoma, undifferentiated carcinoma, bronchial carcinoma, melanoma, renal cell carcinoma, hepatocellular carcinoma, cholangiocarcinoma, cholangiocarcinoma, papillary carcinoma, transitional cell carcinoma, choriocarcinoma, seminal carcinoma, embryonal carcinoma, breast adenocarcinoma, gastrointestinal cancer, colon cancer, bladder cancer, prostate cancer, and squamous cell carcinoma of the neck and head region; e.g., fibrosarcoma, myxosarcoma, sarcoma ... These include sarcomas such as liposarcoma, chondrosarcoma, osteogenic sarcoma, chordal sarcoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, synovial sarcoma, and mesothelial sarcoma; blood cancers such as myeloma, leukemia (e.g., acute myeloid leukemia, chronic lymphocytic leukemia, granulocytic leukemia, monocytic leukemia, lymphocytic leukemia, etc.), lymphoma (e.g., follicular lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma, malignant lymphoma, plasmacytoma, reticulum cell sarcoma, or Hodgkin's disease), and tumors of the nervous system, including glioma, glioblastoma multiforme, meningioma, medulloblastoma, schwannoma, and epididymoma. In one embodiment, the particles of the present invention are adapted for delivery to cancers present as tumors, for example, primary tumors, solid tumors, non-vascularized tumors, and / or tumors of a particular size. Those skilled in the art are knowledgeable as to targeting moieties that can be used to target specific types of cells affected by proliferative disorders, particularly cancer cells.

[0082] In another embodiment, the particles of the present invention do not contain targeting moieties. Whether drug substances are administered to the human body orally, intravenously, or parenterally, they are metabolized by metabolic enzymes in the hepatic portal system (liver), the site of most metabolism, and decompose the delivered substance so that it can be easily removed from the body. Therefore, the particles of the present invention that do not contain targeting moieties can be transported to the liver after being administered to a subject and are well suited for liver treatment. Furthermore, the natural biological distribution of particles can be used to obtain beta-emitting radioisotopes at a given site and irradiate tissue in situ.

[0083] The MOF particles of the present invention contain at least one beta-ray emitting radionuclide. As used herein, the term "radionuclide," which may also be referred to as a radionuclide, radioisotope, or radioactive isotope, is an unstable atom with excess nuclear energy. For beta-ray emitting radionuclides, this excess energy can be utilized in one of three ways: to generate and release new particles (beta particles, beta rays, beta radiation, beta radiation) from the atomic nucleus; or to generate and release new particles (beta particles, beta rays, beta radiation, beta radiation) from the atomic nucleus. During these processes, the radionuclide is said to undergo radioactive decay.

[0084] At least one β-emitting radionuclide is located within the pore of the MOF, e.g., by being present within, e.g., surrounded by, the pore, and is preferably coordinated to and / or chelated by at least one free functional group extending into said pore. Preferably, the β-emitting radionuclide is a β-emitting radionuclide cation.

[0085] Beta particles are high-energy, high-velocity electrons or positrons emitted by the radioactive decay of atomic nuclei during the beta decay process. Beta decay can occur in two forms: beta-decay and beta+ decay, which produce electrons and positrons, respectively. Beta particles with an energy of 0.5 MeV have a range of approximately 1 meter in air. This range depends on the particle energy. Beta particles are a type of ionizing radiation and are considered to have a stronger ionizing effect than gamma rays (useful for imaging) but a weaker effect than alpha particles. Higher ionizing effect results in greater damage to biological tissue, but also results in lower penetrating power of the radiation. The beta-emitting radionuclides used in the present invention have the advantage of having energy levels that are more suitable for deeper tissue penetration than alpha-emitting particles. In one embodiment, the particles of the present invention comprise beta-emitting radionuclides with a tissue penetration depth of approximately 5 mm. Therefore, the beta-emitting particles of the present invention enable new treatments for previously untreatable tumors, such as non-vascular tumors, solid tumors, primary tumors, and / or tumors of certain sizes.

[0086] Beta-emitting radionuclides exert distinct effects on target cells through interference with the cell cycle and specific signal transduction, which can result in tumor regression while minimizing off-target effects on surrounding tissues. Localized delivery of high-energy and highly cytotoxic radiation results in the death of unwanted cells. Radionuclides play a prominent role in inducing apoptosis and cell cycle arrest, as well as improving other characteristics of cancer cells. The application of beta-emitting radionuclides to cancer treatment is emerging as a promising therapeutic approach. Different outcomes can be achieved in cancer cells depending on the radiation dose, exposure time, and type of beta-emitting element. The physical properties of the beta-emitting element (half-life, tissue penetration range, and maximum energy) and the treatment protocol determine the extent to which tumor cells undergo cell cycle arrest, apoptosis, or both.

[0087] In one embodiment, the beta-emitting radionuclide is selected from the group comprising beta-emitting isotopes having beta-excitation energies in the range of 0.3 MeV to 3.5 MeV, or more preferably in the range of 0.5 MeV to 2.3 MeV.

[0088] In one embodiment, the beta-emitting radionuclide has a half-life in the range of 2 hours to 150 days, or more preferably in the range of 24 hours to 51 days.

[0089] In one embodiment, the radionuclide is not terbium 149 or terbium 161. Thus, in one embodiment, terbium 149 and terbium 161, as disclosed in WO 2023 / 282769, are excluded from the group of beta emitters included in the particles of the present invention.

[0090] More preferably, the beta-ray emitting radionuclide of the particles of the invention is selected from the group comprising or consisting of sodium-24, phosphorus-32, potassium-42, calcium-47, scandium-47, iron-59, copper-64, copper-67, strontium-89, yttrium-90, molybdenum-99, ruthenium-103, indium-132, cerium-141, samarium-153, dysprosium-165, holmium-166, erbium-169, thulium-170, lutetium-177, tantalum-182, rhenium-186, rhenium-188, iridium-192, gold-198.

[0091] In some embodiments, the radionuclide is a radionuclide cation, wherein the radionuclide is selected from the group including calcium-47, scandium-47, copper-67, strontium-89, yttrium-90, samarium-153, holmium-166, and lutetium-177, and more preferably, the radionuclide is scandium-47, yttrium-90, or lutetium-177.

[0092] The radionuclides described above have half-lives compatible with tumor killing and clinical administration, and therefore have great therapeutic potential, as set forth below for each isotope in Table 1. These beta-emitting cationic radionuclides described above can bind strongly to the negatively charged chelating groups of MOFs, thereby limiting leakage of the radioisotopes that would otherwise occur in vivo due to competition with salts and other components.

[0093] Table 1 further provides the primary energy levels for each beta emitter.

[0094] [Table 1] TIFF2026501762000005.tif235112 TIFF2026501762000006.tif206112

[0095] In some embodiments, the particles comprise one type of beta-emitting radionuclide.

[0096] In other embodiments, the particles comprise more than one radionuclide, e.g., two, three, e.g., five, where at least one is independently selected from the group provided above. In one embodiment, when the particle population comprises more than one radionuclide, at least one radionuclide is a disclosed beta-emitting radionuclide, and one or more other radionuclides are selected from the group of alpha-emitting radionuclides. Such alpha-emitting radionuclides may be selected from the group including radium-223, radium-224, radium-225, bismuth-212, bismuth-213, lead-212, actinium-225, and thorium-227. Particles of the present invention comprising both beta-emitting and alpha-emitting radionuclides have great potential for enabling treatment of cancerous tissue at different depths, emitting highly toxic alpha particles at the outer layers of tumors while allowing deeper penetration (beta particles) into the cancerous tissue.

[0097] When a particle contains more than one radionuclide, the radionuclides are typically located in separate pores, although the presence of two radionuclides in the same pore may also be possible. It should be noted that in all embodiments, perfect control of the number of radionuclides per particle cannot be expected. During the chelation process, in which the particles are contacted with a solution containing radionuclides, pores of radionuclides per particle will be generated.

[0098] Furthermore, MOFs of the present invention that are suitable for carrying powerful radioisotopes, i.e., beta-emitters optionally combined with alpha-emitters, for use in therapy may also be useful as carriers for short-lived radionuclides for use in imaging. Using the same MOF carrier for different radioisotopes for imaging and therapy ensures nearly identical biodistribution of the MOF carrier containing the imaging agent (e.g., a gamma-emitter) and the therapeutic agent, and this approach allows for pretreatment patient screening to predict response to therapy and avoid unnecessary treatment of non-responders. This universal loading concept and process allows for the use of a single particle platform for both imaging and therapy, obviating the need for separate particle development for each purpose. Thus, the MOFs disclosed herein have been found to be useful as carriers of radionuclides for either therapy or imaging. Thus, the disclosed particles or compositions comprising same can be used in methods of therapeutic procedures comprising the administration of such therapeutic particles, where such treatment is followed by an imaging procedure comprising the use of an imaging agent comprising the same MOF as in the therapeutic particles, but carrying a gamma-ray emitter.

[0099] The particles described herein may have at least the following advantages over the prior art: (a) In contrast to existing small molecule chelators, MOF particles contain a large number of chelating sites, which may enable dose control by loading the particles with a predetermined number of radionuclides. (b) The network structure of MOFs can positively influence the ability of the particles to contain radiation, for example, by the following mechanisms: (1) In contrast to state-of-the-art molecular chelators, MOFs can contain, by physical adsorption, daughter isotopes that emerge through the decay chain of radionuclides and thus also retain the daughter isotopes in the vicinity of tumor cells. (2) In contrast to state-of-the-art molecular chelators, radionuclides adsorbed on MOFs are less susceptible to competitive adsorption by other ions because they can be adsorbed by the defective pores. (3) The desorbed radionuclide can be readsorbed into adjacent deletion pores.

[0100] In summary, the present invention may therefore provide improved methods of radiation therapy using beta-emitters.

[0101] Regarding (b)(2), competitive adsorption may be relevant not only during particle preparation and storage, and during adsorption of radionuclide cations, but also when the particles are used as pharmaceuticals. For example, Sr 2+ is generally Mg 2+ and Ca 2+ However, the use of MOFs is less susceptible to competition from Sr than the use of low-molecular-weight chelating agents. 2+ See the results of Gao et al. and Example 7, which show that PEG-1000 absorbed and retained Lu very well for several days.

[0102] The particles of the present invention may further comprise one or more molecules for modifying the particle's exterior surface. Such one or more molecules may be linked to the particle, e.g., coordinated, e.g., covalently bonded, etc., at the particle's exterior surface. Again, size discrimination may be used to conduct reactions on the exterior surface rather than within the pores. For example, the exterior surface of a MOF containing free carboxyl or amino groups may be functionalized with one or more compounds selected from the group consisting of PEG derivatives, N-hydroxysuccinimide (NHS), N-hydroxysulfosuccinimide (sulfo-NHS), MD linkers, Mal-PAB, and albumin to facilitate transport, prevent aggregation, and / or provide targeting functionality by increasing targeting flexibility and spatial distance between the MOF and its target. Albumin may be used to prevent aggregation and increase the particle's blood half-life.

[0103] The particles of the invention may further comprise one or more additional compounds, such as molecules, e.g., ions, located in at least one of the pores. Non-limiting examples include buffers and / or specific ions, e.g., ions to limit leakage of radionuclides due to steric hindrance.

[0104] MOF particles are typically produced by aqueous synthesis at 20-100°C, where solutions of appropriate inorganic precursors for inorganic monomers and organic monomers are mixed. In some cases, the organic monomers are dissolved directly in the inorganic precursor solution, or vice versa. In some cases, growth control factors (also called growth modulators) are added to influence the particle growth rate. MOF particles precipitate from solution as the structure forms. Published procedures for MOF production can be followed.

[0105] Targeting moieties can be attached, linked, attached, commonly referred to as conjugation, to the MOF particle directly or via a linker. For example: carboxyl MOFs can be conjugated to amine groups (present on the targeting unit) by carbodiimide (EDC, NHS / sulfo-NHS)-based crosslinking.

[0106] Radiolabeling can be carried out by uniformly mixing a solution or suspension of radionuclide cations with a suspension of unlabeled particles for more than 1 minute, and then separating the remaining unbound radionuclide cations from the labeled particles, for example, by centrifugation or column purification. If the targeting site is present on the particle before radiolabeling is performed, the radiolabeling procedure can be more convenient, for example, in terms of time used, purification of the product, etc.

[0107] Thus, the particles of the present invention can be prepared by a process comprising providing a particle, e.g., a nanoparticle, comprising a MOF, the process comprising: where MOF is a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; wherein the particles optionally comprise At least one targeting moiety attached to the particle on the outer surface of the particle. and contacting the particles with a beta-ray emitting radionuclide, e.g., a radionuclide cation, wherein the radionuclide may be selected from the group including sodium-24, phosphorus-32, potassium-42, calcium-47, scandium-47, iron-59, copper-64, copper-67, strontium-89, yttrium-90, molybdenum-99, ruthenium-103, indium-132, cerium-141, samarium-153, dysprosium-165, holmium-166, erbium-169, thulium-170, lutetium-177, tantalum-182, rhenium-186, rhenium-188, iridium-192, and gold-198; It can be prepared by

[0108] The radionuclide may be present in a composition that includes, for example, a liquid.

[0109] One aspect of the invention is a method for preparing particles by homogeneously mixing a solution or suspension of a β-emitting radionuclide with a suspension of particles comprising a MOF, comprising: The MOF is a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the free functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; and thereafter separating any remaining unbound beta-emitting radionuclides from the labeled particles. The present invention provides a method comprising:

[0110] In another aspect, the present invention relates to a kit, said kit comprising: In the first container, a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs containing; Optionally, at least one targeting moiety linked to the particle on the outer surface of the particle; and In the second container, Beta-emitting radionuclides Includes:

[0111] The beta-emitting radionuclide may be selected as disclosed in the first alternative, and thus may be selected from the group of sodium-24, phosphorus-32, potassium-42, calcium-47, scandium-47, iron-59, copper-64, copper-67, strontium-89, yttrium-90, molybdenum-99, ruthenium-103, indium-132, cerium-141, samarium-153, dysprosium-165, holmium-166, erbium-169, thulium-170, lutetium-177, tantalum-182, rhenium-186, rhenium-188, iridium-192, and gold-198.

[0112] Those skilled in the art will understand that the first container and the second container may further comprise a liquid, such as a solvent, for dissolving or suspending any components, as well as additional components, such as at least one carrier, diluent, and / or excipient.

[0113] The particles disclosed herein, i.e., at least one particle, and preferably a plurality of particles, can be present as an active ingredient in a desired dosage unit formulation, such as a pharmaceutically acceptable composition containing a conventional pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable" means that the compound must be physiologically tolerable to the recipient and, when part of a composition, compatible with the other components of the composition. The term "composition" refers to a mixture of one or more compounds according to the present invention with one or more additional chemical components in any formulation.

[0114] Thus, in another aspect, the present invention relates to a composition comprising at least one particle as disclosed in the first aspect, together with at least one pharmaceutically acceptable carrier, diluent and / or excipient.

[0115] The particle is defined as disclosed above. It may be a microparticle or a nanoparticle. Preferably, the particle is a nanoparticle.

[0116] The composition may be considered a pharmaceutical composition because it comprises an active agent, i.e., at least one particle, in combination with at least one pharmaceutically acceptable carrier, diluent and / or excipient, making the composition particularly suitable for therapeutic use.

[0117] The composition preferably comprises a plurality of particles of the present invention. The particles may be the same or different, i.e., with respect to the type and number of radionuclides and / or targeting moieties. In some embodiments, the composition is a particle suspension comprising monodisperse or polydisperse particles labeled with radionuclide cations.

[0118] The compositions may contain one or more of any conventional, pharmaceutically acceptable excipients and / or carriers, such as solvents, fillers, diluents, binders, lubricants, glidants, viscosity adjusters, surfactants, dispersants, disintegrants, emulsifiers, wetting agents, suspending agents, thickening agents, buffers, pH adjusters, absorption delaying agents, stabilizers, antioxidants, preservatives, antibacterial agents, antifungal agents, chelating agents, adjuvants, sweeteners, flavoring agents, and coloring agents, etc. Conventional formulation techniques known in the art, such as conventional mixing, dissolving, suspending, granulating, dragee-making, extruding, emulsifying, encapsulating, entrapping, or compressing processes, may be used to formulate the compositions.

[0119] In some embodiments, the composition is formulated for a particular method of administration to a subject.

[0120] The amount of particles according to the present invention present in the composition can vary. In some embodiments, the amount of particles according to the present invention present in the composition is 0.1 to 50% by weight, for example, 1 to 30% by weight, for example, 20 to 50% by weight. In other embodiments, the amount of particles according to the present invention present in the composition is 30 to 70% by weight, for example, 40 to 60% by weight. In still other embodiments, the amount of particles according to the present invention present in the composition is 50 to 100% by weight, for example, 50 to 70% by weight, for example, 50 to 80% by weight, for example, 60 to 98% by weight, for example, 70 to 95% by weight.

[0121] The composition may also include MOF particles that do not contain radionuclide cations, such as particles containing MOFs, where the MOFs comprise a repeating three-dimensional network of inorganic and organic monomers that form pores and at least one free functional group extending into the pores, where the functional group is selected from the group consisting of carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof, and the particles comprise at least one targeting site linked to the particle on the particle's exterior surface. Such particles may be the same as or different from the particles of the present invention except for the absence of radionuclide cations. The ratio between particles containing radionuclide cations and particles not containing radionuclide cations may vary. In a preferred embodiment, at least 90% of the particles contain radionuclide cations. The activity per mg of particle is typically in the range of 10 kBq to 570,000 MBq, e.g., 100 kBq to 570,000 MBq, or 100 kBq to 1000 MBq.

[0122] Furthermore, the composition is substantially free of contaminants or impurities. In some embodiments, the level of contaminants or impurities other than residual solvent in the composition is less than about 5% by weight based on the total weight of the particles according to the present invention and other intended components. In certain embodiments, the level of contaminants or impurities other than residual solvent in the composition is less than about 2% or 1% by weight based on the total weight of the particles according to the present invention and other intended components.

[0123] In certain embodiments, the particles or compositions according to the invention are sterile. Sterilization can be achieved by any suitable method, including but not limited to, application of heat, chemicals, irradiation, high pressure, filtration, or a combination thereof.

[0124] The particles of the present invention can be included in a composition and used as a pharmaceutical. Thus, the present invention provides the particles disclosed in the first aspect for use as a pharmaceutical. In another aspect, the present invention provides particles for use as a pharmaceutical, comprising: a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, the free functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs containing; Optionally, at least one targeting moiety linked to the particle on the outer surface of the particle; and at least one beta-emitting radionuclide; Including, wherein said at least one radionuclide is located in at least one of the pores of said particle, Or a composition comprising at least one of said particles together with at least one pharmaceutically acceptable carrier, diluent and / or excipient.

[0125] The particles of the present invention and compositions comprising the particles can be used therapeutically, such as for targeted delivery of radioactive decay to one or more specific sites in vivo, such as specific cells, tissues, or organs. The targeting moiety targets the particle to the specific site where radioactive decay, such as beta particle emission, is desired, and the MOF functions as a chelating agent with the advantageous properties described above. If the particle does not contain a targeting moiety, the particle will be transported to other organs, such as the liver, where the radioactivity will decay, depending on the biodistribution properties of the nanoparticles. Thus, in one embodiment, the particles can be used therapeutically for indications where nanoparticles are naturally transported, particularly for the treatment of liver cancer.

[0126] For the various aspects of the invention described herein relating to use as a pharmaceutical and / or for the treatment or diagnosis of disease, particularly the selective targeting of diseased tissue, the diseased tissue may in all embodiments be present at a single site within the body, for example in the case of a localized solid tumor, or may be present at multiple sites, for example in the case of a distributed or metastatic cancerous disease.

[0127] Due to the cytotoxic effect of radioactive decay, the particles and compositions of the present invention may be particularly useful for proliferative diseases.Accordingly, in a further aspect, the present invention relates to the disclosed particles or compositions comprising said particles together with at least one pharmaceutically acceptable carrier, diluent and / or excipient for use in a method for treating proliferative diseases.

[0128] Due to the ability of beta-emitting radionuclides to penetrate deeper into tissue than, for example, alpha-emitting radionuclides, the particles of the present invention or compositions comprising them are particularly suitable for use in the treatment of primary tumors, solid tumors, non-vascular tumors and / or tumors of a certain size, e.g., 0.1 mm or less depending on shape and accessibility. 3 Tumors of this volume and above can be treated according to the present invention.

[0129] Particles are defined as disclosed herein and may be nanoparticles or microparticles, preferably nanoparticles.

[0130] The terms "treating," "treatment," and "therapy" (and grammatical variations thereof) are used interchangeably herein and refer to: 1) inhibiting a disease; for example, inhibiting a disease, condition, or disorder in a subject pathologically or symptomatically experiencing or manifesting the disease, condition, or disorder, including preventing disease (i.e., prophylactic treatment, preventing further progression of the disease and / or symptoms); or 2) alleviating the symptoms of a disease; or 3) ameliorating a disease; for example, ameliorating a disease, condition, or disorder in a subject pathologically or symptomatically experiencing or manifesting the disease, condition, or disorder (i.e., reversing the pathology and / or symptoms). Preferably, treatment refers to the amelioration of a disease, for example, by killing tumor cells, inducing apoptosis, and / or inhibiting cancer cell proliferation. These terms may relate to the use and / or administration of pharmaceuticals, active pharmaceutical ingredients (APIs), and / or pharmaceutical-grade supplements.

[0131] As used herein, the terms "administer," "administration," and "administering" refer to (1) providing, giving, dispensing, and / or prescribing a formulation, preparation, or composition according to the present disclosure, either by or under the direction of a healthcare professional or their authorized representative, or by self-administration, and (2) the subject administering, ingesting, or consuming a formulation, preparation, or composition according to the present disclosure.

[0132] As used herein, "subject" means a human or non-human animal selected for treatment or therapy, and includes, and may be limited to, a "patient." Neither term should be construed as requiring the supervision (full or otherwise) of a medical professional (e.g., physician, nurse, nurse practitioner, physician assistant, orderly, clinical research associate, etc.) or a scientific researcher.

[0133] The subject is preferably a human subject. The subject may be male or female. In some embodiments, the subject is an adult (i.e., 18 years of age or older). In certain embodiments, the subject is elderly. In certain embodiments, the subject is not elderly. The subject is preferably a subject diagnosed with a proliferative disease, such as cancer.

[0134] The targeted diseased tissue may be a soft tissue site, a calcified tissue site, or multiple sites that are all soft tissue, multiple sites that are all calcified tissue, or may include at least one soft tissue site and / or at least one calcified tissue site. In one embodiment, at least one soft tissue site is targeted. The targeted site and the site of origin of the disease may be the same or different. When multiple sites are involved, this may include the site of origin or multiple secondary sites. The term "soft tissue" is used herein to refer to tissue that does not contain a "hard" mineralized matrix. In particular, "soft tissue" as used herein may be any tissue that is not skeletal tissue. Correspondingly, "soft tissue disease" as used herein refers to a disease that occurs in "soft tissue" as used herein. The present invention is particularly suited to the treatment of cancer, and thus "soft tissue disease" encompasses carcinomas, sarcomas, myelomas, leukemias, lymphomas, and mixed cancers arising in any "soft" (i.e., non-mineralized) tissue, as well as other non-cancerous diseases of such tissue. Cancerous "soft tissue disease" includes solid tumors arising in soft tissue, as well as metastatic tumors and micrometastatic tumors, and the present invention is particularly suited for the treatment of solid tumors. Indeed, a soft tissue disease may consist of a primary solid tumor of soft tissue and at least one metastatic tumor of soft tissue in the same patient. Alternatively, a "soft tissue disease" may consist solely of a solid tumor whose primary tumor is a skeletal disease, or solely of metastatic tumors.

[0135] When the particles or compositions of the present invention are used as pharmaceuticals and / or in therapeutic methods according to the present invention, target sites may be selected based on specific biomarkers, such as antigens, expressed by or in the vicinity of tissues, cells or organs affected by a proliferative disease.

[0136] In some embodiments, the proliferative disorder is a cancer, a non-cancerous tumor, a neoplastic disorder, or a hyperplastic disorder.

[0137] In some preferred embodiments, the proliferative disease is cancer. As used herein, the terms "cancer" and "tumor" refer to any neoplastic growth in a subject, including primary tumors and metastases. Cancer can be of liquid or solid tumor type. Liquid tumors include tumors of hematological origin, including, for example, myeloma (e.g., multiple myeloma), leukemia (e.g., Waldenstrom syndrome, chronic lymphocytic leukemia, other leukemias), and lymphomas (e.g., B-cell lymphoma, non-Hodgkin's lymphoma). Solid tumors can occur in organs, including, but not limited to, cancer of the lung, brain, breast, prostate, ovary, colon, kidney, and liver.

[0138] In some embodiments, the cancer is selected from the list comprising or consisting of lung cancer, pancreatic cancer, colorectal cancer; liver cancer, glioma, renal cancer, non-Hodgkin's lymphoma, neuroblastoma, CNS metastasis, peritoneal cancer, follicular lymphoma, colorectal cancer, small cell lung cancer, carcinoma, sarcoma, myeloma, leukemia, lymphoma, prostate cancer or mixed cancer.

[0139] In certain embodiments, the cancer comprises a solid, non-vascularized tumor, which can be difficult to treat using conventional anti-cancer drug therapies, and the targeted MOF vehicles of the present invention loaded with beta-emitters represent a promising line of treatment for such cancers.

[0140] The targeting moiety of the particle will be selected based on the specific disease to be treated. For example, CD37 is highly expressed on B cells and the majority of B cell lymphomas, is absent on normal stem cells, and disappears again upon differentiation into plasma cells. Due to its high prevalence on the surface of B lymphomas, CD37 has become the target of several different drugs in clinical development. Therefore, anti-CD37 may be a useful targeting moiety for such cancers.

[0141] Non-Hodgkin's lymphoma spreads mainly through the lymphatic system, making it difficult to treat with conventional therapies (surgery, radiation). Furthermore, non-Hodgkin's lymphoma can metastasize to other tissue types, which is another reason why the present invention may be particularly effective against non-Hodgkin's lymphoma.

[0142] The particles or compositions for use in pharmaceuticals and / or therapeutic methods according to the present invention are administered to a subject at a therapeutically effective dose. As used herein, the term "therapeutically effective dose" refers to an amount of particles according to the present invention that is effective to produce a desired therapeutic effect in a subject at a reasonable benefit / risk ratio applicable to any treatment. The therapeutically effective dose may vary depending on the route of administration and dosage form. Furthermore, the dosage may depend on the particles used, the stage of symptoms, the age and weight of the subject, etc., and can be routinely determined by those skilled in the art according to principles well known in the art.

[0143] In some embodiments, the amount of radionuclide cation used per patient dose may be in the range of 1 MBq to 100 GBq, preferably 1 MBq to 50 MBq, more preferably 10 MBq to 25 GBq, and even more preferably 100 MBq to 10 GBq. Dosages and maximum doses can be determined by those skilled in the art based on general knowledge of appropriate dosages and maximum doses. It is recognized in the art that realistic and conservative estimates of toxic side effects of daughter isotopes must be employed.

[0144] In some embodiments, the particles are administered at a dose of 10 kBq to 1 GBq / kg body weight, e.g., 10 kBq to 500 MBq / kg body weight, e.g., 100 kBq to 250 MBq / kg body weight, preferably 1 MBq to 100 MBq / kg, e.g., 10 MBq to 100 MBq / kg, more preferably 50 MBq to 100 MBq / kg, and particularly 50 MBq to 80 MBq / kg. Correspondingly, a single dosage unit may contain anywhere in this range (e.g., a range of 1 MBq to 100 GBq per dosage) multiplied by the appropriate body weight, e.g., 30 to 150 kg, preferably 40 to 100 kg. The dosage, particles and route of administration may be such that the dose of progeny nuclides produced in vivo is less than 300 kBq / kg, for example less than 200 kBq / kg, preferably less than 150 kBq / kg, for example less than 100 kBq / kg.

[0145] A therapeutically effective amount of the particles or compositions of the present invention can be administered in a single dose or in divided doses. The particles or compositions of the present invention can be administered once a day, twice or more times a day, every other day, every three days, twice a week, or once a week, or as deemed appropriate by a medical professional. In certain embodiments, the particles or compositions of the present invention are administered once a day. In other embodiments, the particles or compositions of the present invention are administered twice a day. In some embodiments, the administration regimen is predetermined and is the same for the entire patient population. In other embodiments, the dosage and frequency of treatment with the particles or compositions of the present invention will be determined by a medical professional based on factors including, but not limited to, the stage of the disease, the severity of symptoms, the route of administration, the subject's age, weight, general health, sex, and / or diet, and / or the subject's response to treatment.

[0146] In some embodiments, the therapeutically effective dose is administered at regular intervals. In other embodiments, the dose is administered as needed or sporadically. The particles or compositions according to the present invention can be administered by a medical professional. The particles or compositions according to the present invention can be administered with or without food, depending on factors such as the formulation and the route of administration. In some embodiments, the particles or compositions according to the present invention are administered at a specific time of day.

[0147] The particle or composition for use in the pharmaceutical and / or therapeutic method according to the present invention can be administered locally or systemically.The particle or composition according to the present invention can be administered by any administration route, including but not limited to pulmonary administration, oral administration, intraperitoneal administration, intravenous administration, intramuscular administration, intratumoral administration, sublingual administration, subcutaneous administration, intrahepatic administration, buccal administration, rectal administration, intravaginal administration, occipital administration, intranasal administration, transdermal administration and intradermal administration.In one embodiment, administration is selected from the group of intraperitoneal administration, intravenous administration, intramuscular administration, intratumoral administration and subcutaneous administration, and preferably intravenous administration.

[0148] In some embodiments, the particles or compositions according to the present invention are administered intravenously. In these embodiments, water is a particularly useful excipient. Saline solutions and aqueous solutions of dextrose and glycerol can also be employed as liquid excipients, particularly for injectable solutions.

[0149] A preferred dosage formulation comprises the therapeutically effective dose or a suitable fraction thereof of the particles according to the present invention as described hereinabove.The composition of the present invention can be presented in a unit dosage form as a single dose, in which all active and inactive ingredients are combined in a suitable system and do not need to be mixed before administration.Alternatively, the composition can be presented as a kit as disclosed above, and can include instructions for storing, preparing, administering and / or using the composition.

[0150] In some embodiments, the duration of use of a particle or composition for use as a pharmaceutical and / or in a method of treatment according to the present invention is determined by the observed therapeutic effect, e.g., reduction and / or amount of target antigen expression. In some embodiments, treatment is continued until no further improvement is expected. In certain embodiments, the duration of treatment with a particle or composition according to the present invention is at least 2 weeks, at least 1 month, or at least 3 months, e.g., 3 months, 6 months, 9 months, 1 year, 3 years, or 5 years. In other embodiments, the duration is determined by a medical professional based on factors including, but not limited to, the nature and severity of the symptoms, the route of administration, the subject's age, weight, general health, sex, and / or diet, and / or the subject's response to treatment.

[0151] In certain embodiments, the particles or compositions of the present invention are administered alone. In other embodiments, the particles or compositions of the present invention are administered in combination with one or more other therapeutic agents. The one or more other therapeutic agents may be known to have an effect on a proliferative disease, such as cancer, and / or may have an additive or synergistic mechanism of action with the particles or compositions of the present invention in treating the proliferative disease, such as cancer. In some embodiments, the particles or compositions of the present invention are administered as part of a combination therapy. A combination therapy including the particles or compositions of the present invention can refer to a composition comprising the particles or compositions of the present invention in combination with one or more therapeutic agents, and / or a composition in which the particles or compositions of the present invention and one or more therapeutic agents are co-administered, where the particles or compositions of the present invention and the other therapeutic agents or drugs are not formulated in the same composition. When separate formulations are used, the particles or compositions of the present invention can be administered simultaneously, intermittently, staggered, before, after, or a combination thereof with the administration of another therapeutic agent.

[0152] Embodiments and features described in the context of one aspect, e.g., directed to particles or compositions, also apply to other aspects of the invention, e.g., their use as medicaments or in methods of treatment.

[0153] In a further aspect, the present invention provides a method of treatment, the method comprising administering to a subject in need thereof an effective amount of a particle or composition of the present invention.

[0154] In a further aspect, the present invention provides a method of treating a proliferative disease, the method comprising administering to a subject in need thereof an effective amount of a particle or composition of the present invention.

[0155] In a further aspect, the present invention provides a method for treating a chronic inflammatory disease, the method comprising administering to a subject in need thereof an effective amount of a particle or composition of the present invention.

[0156] In a further aspect, the present invention provides the use of a particle or composition of the invention as a medicament.

[0157] In a further aspect, the present invention provides the use of a particle or composition of the invention for the treatment of a proliferative disorder.

[0158] In a further aspect, the present invention provides the use of a particle or composition of the invention for the treatment of a chronic inflammatory disease.

[0159] The present invention is not limited to the illustrated embodiments and examples. While various embodiments of the present disclosure have been described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Numerous modifications and variations, as well as variations and substitutions to the embodiments described herein, will be apparent to those skilled in the art without departing from the present disclosure. It will be understood that various alternatives to the embodiments described herein may be employed in practicing the present disclosure.

[0160] It is to be understood that all embodiments of the present disclosure may be combined in any manner with any one or more of the other embodiments described herein.

[0161] It is understood that each component, compound, particle, or parameter disclosed herein is intended to be disclosed for use alone or in combination with one or more of the other components, compounds, or parameters disclosed herein. Furthermore, it is understood that each amount / value or amount / value range of each component, compound, or parameter disclosed herein is intended to be disclosed in combination with each amount / value or amount / value range disclosed for any other component, and any combination of amount / values ​​or amount / value ranges for two or more components, compounds, or parameters disclosed herein is therefore intended to be disclosed in combination with each other for purposes of this specification. Any and all features described herein, and combinations of such features, are included within the scope of the present invention, provided that the features are not mutually inconsistent.

[0162] It should be understood that each lower limit of each range disclosed herein is interpreted as a disclosure in combination with each upper limit of each range disclosed herein for the same component, compound, or parameter. Thus, a disclosure of two ranges is interpreted as a disclosure of four ranges derived by combining each lower limit with each upper limit of each range. A disclosure of three ranges is interpreted as a disclosure of nine ranges derived by combining each lower limit with each upper limit of each range. Furthermore, a specific amount / value of a component, compound, or parameter disclosed in the specification or examples is interpreted as a disclosure of either the lower or upper limit of a range, and therefore may be combined with other lower or upper limits, or ranges or specific amounts / values, of the same component, compound, or parameter disclosed elsewhere in this application to form a range for that component, compound, or parameter. [Example]

[0163] Example 1: Conjugation of anti-CD37 IgG to UIO-66-(COOH) UIO-66-(COOH)2 MOF was prepared by a method based on that of Zhiijie Chen et al., CrystEngComm 2019, 14, 2409-2415, except that the oxychloride salt was used as the precursor for the MOF instead of the oxynitrite salt. UIO-66-(COOH)2 antibody conjugation was evaluated by analyzing UIO-66-(COOH)2 antibody-conjugated particles using the bicinchoninic acid assay (BCA). The BCA protein assay measures Cu by protein in alkaline medium. 2+ Cu 1+ The well-known reduction of copper cations (Cu) to 1+ The first step is the chelation of copper with proteins in an alkaline environment, forming a pale blue complex. In this reaction, known as the biuret reaction, peptides containing three or more amino acid residues form a colored chelate complex with copper ions in an alkaline environment containing sodium potassium tartrate.

[0164] In the second step of the color reaction, BCA reacts with the reduced cation (cuprous ion) formed in the first step. A deep purple reaction product results from the chelation of two molecules of BCA with one cuprous ion.

[0165] procedure: Five mg of UIO-66-(COOH)2 was suspended in 0.05 M 2-morpholinoethanesulfonic acid (MES) buffer, pH 6.3, and treated with 1.2 mg of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and 1.2 mg of sodium N-hydroxysulfosuccinimide (sulfo-NHS) for 15 minutes. The particles were then washed twice with 0.05 M MES buffer, pH 6.3. The particles were then resuspended in 0.05 M MES buffer, pH 6.3, containing 100 μg of anti-CD37 IgG. The reaction mixture was incubated for 3 hours with continuous rocking. The MOF-antibody particles were washed with 0.05 M MES buffer, pH 6.3, and resuspended in 100 mM Tris buffer. The particles were then washed and stored in 0.05 M MES buffer, pH 6.3.

[0166] Antibody (IgG)-conjugated UIO-66-(COOH)2 and albumin standards were evaluated by the BCA assay shown in Figure 12. The albumin trend line equation was used to calculate the protein concentration of UIO-66-(COOH)2-IgG particles per mL: absorbance-blank: 0.096, equivalent to 77 μg / mL (77% of the antibody input amount).

[0167] Other MOFs suitable for use in the present invention, such as UIO-66-(COOH), MOF-808, or UIO-66-(COOH)2 types, such as Hf-UiO-66, Ce-UiO-66, and MOF-808, can be prepared by similar procedures and can be further tested for stability, adsorption and retention capacity, binding to target molecules, and therapeutic uses, as shown in the examples below.

[0168] Example 2: Structural stability and barium retention capacity of MOFs in human serum In this experiment, barium(II) was used to demonstrate the structural stability and retention ability of the labeled MOF. Ba(II) and Sr(II) have similar ionic radii (Sr 2+ 132pm, Ba2+ They have similar orbital structures (both are members of the alkaline earth metal group and have the electron configuration of a noble gas in an adjacent period), and are expected to behave similarly in chemical and biological systems.

[0169] procedure: 200 mg of UIO-66-(COOH)2 was washed once with 0.05 M MES buffer (pH 6.3) and suspended in 40 mL of aqueous barium(II) acetate (14.7 μg Ba / mL). The suspension was stirred at room temperature for 60 min. The MOF was recovered, washed once with 5 mL of 0.05 mol / L MES buffer (pH 6.3), and then suspended in human serum. The MOF / serum suspension was stirred at room temperature for 11 days. Samples were taken after 15 min and after 1, 2, 4, and 7 days. Elemental analysis was performed using MP-AES after digestion of the serum sample with a 1:1 volume mixture of HNO3 (65%) aq. and H2O2 (33%) aq. at 70 °C for 1 h.

[0170] Elemental analysis showed that the MOF adsorbed 97.2% of the Ba2+ in solution, with a Ba2+ concentration of 2.87 μg / mg (0.287% wt.) in the MOF. Analysis of serum samples taken after 15 min and 1, 2, 4, and 7 days indicated a total Ba2+ leakage into the serum of 6.6%, 8.4%, 9.4%, 8.3%, and 5.5%, respectively. This indicates that after the initial leakage, the MOF contained all of the Ba2+.

[0171] The MOF was isolated from the serum after 11 days and analyzed by powder X-ray diffraction and energy dispersive X-ray spectroscopy. The powder X-ray diffraction patterns of UIO-66-(COOH)2 as received and after the 11-day stability test clearly show that the crystalline structure of the MOF remains intact. EDS indicates that the Ba / Zr mass ratio in the MOF is 1.10%, which is comparable to the theoretical mass fraction of Zr in the MOF (25.1% from the formula Zr6O4(OH)4(COO)12(C8H4O4)6, which is 1.14%, and is in close agreement with the value measured by MP-AES (0.287% wt.), within experimental error.

[0172] [Table 2]

[0173] Therefore, this experiment demonstrates that the MOF particles themselves are stable in human serum for at least 11 days, and that the chelation of barium cations in the MOF particles is stable for at least 11 days. This experiment also demonstrates the ability of MOFs to adsorb significant amounts of Ca, K, and Na without leaking Ba.

[0174] Example 3: Structural stability and barium retention capacity of MOFs in human serum This example confirms the results of Example 2 using a different Ba2+ source and two different concentrations.

[0175] procedure: Two separate portions of 120 mg of UIO-66-(COOH)2 were washed three times with 5 mL of 0.05 mol / L MES buffer (pH 6.3) and then suspended in two separate 5 mL aqueous solutions of barium(II) nitrate (1.165 mg / mL Ba and 0.109 mg / mL Ba, respectively). The suspensions were stirred at room temperature for 90 min. The MOFs were recovered, washed once with 5 mL of 0.05 mol / L MES buffer (pH 6.3), and then suspended in human serum. Samples were taken after 24 and 48 h. The MOF / serum suspension was continued stirring at room temperature for 2 days. Samples were taken after 24 and 48 h. Elemental analysis was performed using MP-AES after digestion of the serum samples with a 1:1 volume mixture of HNO3 (65%) aq. and H2O2 (33%) aq. at 70 °C for 1 h.

[0176] From a 1.165 mg / mL Ba solution: Elemental analysis showed that the MOF adsorbed 98.8% of the Ba2+ in solution, resulting in a Ba2+ concentration of 48.0 μg / mg (4.8% wt.) in the MOF. Analysis of serum samples taken after 24 and 48 h showed a total Ba2+ leakage into the serum of 8.8% and 9.2%, respectively. This indicates that there was almost negligible Ba2+ leakage between 24 and 48 h. EDS showed that the Ba / Zr mass ratio in the MOF was 19.4%, which is comparable to the theoretical mass fraction of Zr in the MOF (25.1% from the formula Zr6O4(OH)4(COO)12(C8H4O4)6), and is in close agreement, within experimental error, with the value measured by MP-AES (4.8% wt.).

[0177] From a 0.109 mg / mL Ba solution: Elemental analysis showed that the MOF adsorbed 92.0% of the Ba2+ in solution, resulting in a Ba2+ concentration of 4.19 μg / mg (0.42% wt.) in the MOF. Analysis of serum samples taken after 24 and 48 hours showed a total Ba2+ leakage into the serum of 6.4% and 6.0%, respectively. This indicates that there was no Ba2+ leakage between 24 and 48 hours. EDS showed that the Ba / Zr mass ratio in the MOF was 1.43%, compared to the theoretical mass fraction of Zr in the MOF (25.1% from the formula Zr6O4(OH)4(COO)12(C8H4O4)6 of 1.67%, which is in close agreement, within experimental error, with the value measured by MP-AES (0.42% wt.).

[0178] [Table 3]

[0179] Example 4: MOF-IgG cell interactions This example demonstrates that anti-CD37 conjugated MOF (NP1) binds to CD37-positive cells (Daudi cells). The Daudi cell line consists of B lymphoblasts isolated from the peripheral blood of a patient with Burkitt's lymphoma, demonstrating that anti-CD37 conjugated MOF can identify blood-borne cancer cells.

[0180] procedure: Sixty-six thousand Daudi cells were transferred to flow tubes containing phosphate-buffered saline (PBS), an isotype control, a UIO-66-(COOH)2 anti-CD37 stability test sample (incubated at 20°C for 6 months), or a freshly conjugated sample of UIO-66-(COOH)2 anti-CD37. The samples were incubated for 30 minutes with continuous mixing, after which 400 μL of 10% fetal bovine serum (FBS) in PBS was added and centrifuged at 350×g for 8 minutes. The supernatant was discarded, and then 100 μL of anti-CD37-PE (10 μL of anti-CD37-PE mixed with 90 μL of PBS) was added to the PBS samples, or 100 μL of anti-mouse-PE (10 μL of anti-mouse-PE mixed with 90 μL of PBS) was added to the isotype control and UIO-66-(COOH)2a CD37 samples. The samples were incubated for 30 minutes with continuous mixing, after which 400 μL of 10% FBS in PBS was added and centrifuged at 350 g for 8 minutes, the supernatant was discarded, and then 400 μL of 10% FBS in PBS was added and flow cytometry analysis was performed.

[0181] The stability control sample used in this experiment was prepared by storing UIO-66-(COOH)2 anti-CD37 at 20°C for 6 months.

[0182] FIG. 13 provides results showing that anti-CD37 conjugated MOF (NP1) binds to CD37-positive cells and is stable at room temperature for at least 6 months.

[0183] Example 5: Cellular binding of MOF-anti-EpCAM and MOF-anti-HER2 to colorectal cell lines This example demonstrates that UIO-66-(COOH)2 conjugated to either anti-EpCAM or anti-HER binds to colon cancer cells (HTC116 and HT29) expressing both antigens. HTC116 cells were derived from the colon of an adult male patient with colon cancer. HT29 cell line was derived from the primary tumor of a female patient with colon adenocarcinoma. Therefore, we demonstrate that UIO-66-(COOH)2 conjugated to both anti-EpCAM and anti-HER antibodies (IgG) can identify cancer cells derived from the colon.

[0184] procedure: Cells were seeded at 50,000 cells per well in a 96-well plate and cultured overnight in Roswell Park Memorial Institute (RPMI) complete cell culture medium supplemented with 10% fetal bovine serum (FBS). The next day, the medium was discarded from each well, and 2 μg of anti-EpCAM, 2 μg of isotype control, or 15 μL of UIO-66-(COOH)2 (NP1) diluted in prewarmed cell culture medium was added to a total volume of 200 μL per well. Samples were incubated at 37°C for 30 minutes, after which the medium was discarded and 200 μL of phosphate-buffered saline (PBS) was gently added twice. 100 μL of PBS and 100 μL of anti-human IgG-HRP solution (2.5 μL anti-human IgG-HRP solution / mL) were added per sample and incubated at room temperature for 30 minutes. After secondary staining, cells were gently washed twice with PBS, and then 100 μL of TMB (3,3',5,5'-tetramethylbenzidine) HRP (horseradish peroxidase) substrate solution was added to each well and incubated for 15 minutes. 100 μL of stop solution was added per well to stop the reaction before measuring the absorbance of each well at 450 nm. The absorbance is displayed on the graph as "HRP signal."

[0185] FIG. 14 shows that anti-EpCAM and anti-HER2 UIO-66-(COOH) 2 bind colon cancer cells (HTC116 and HT29).

[0186] Example 6: MOF-anti-HER2 breast cancer cell interactions This experiment demonstrates that anti-HER2 conjugate UIO-66-(COOH)2 binds HER2-positive cancer cells (JIMT1 cells). JIMT1 cells are epithelial cells derived from a female patient with breast ductal adenocarcinoma. Therefore, the interaction with JIMT1 cells demonstrates how anti-HER2 conjugate UIO-66-(COOH)2 can identify cancer cells derived from the breast.

[0187] Procedure: The same procedure as for colon cancer cell interaction was used for JIMT1 cell interaction, see Example 5 (Figure 14).

[0188] FIG. 15 shows that anti-HER2 conjugate UIO-66-(COOH)2 binds to breast cancer JIMT1 cells.

[0189] Example 7: Adsorption and retention capacity of MOFs for β-emitters in human serum The adsorption and retention capacity of β-emitters onto MOFs, namely lutetium-177 (Lu177) onto UIO-66(COOH)2, was evaluated. Samples were incubated in serum, which represents a suitable in vitro environment for assessing isotope binding.

[0190] Procedure: Radiolabeling is performed using 60 kBq [ 177 The assay was carried out by adding a solution of [Lu]LuCl3 to a 400 μL suspension of UIO-66(COOH)2 (0.6 mg) in 0.005 mol / L MES buffer (pH 6). 177A control sample consisting of [Lu]LuCl3 was prepared in parallel. The samples were kept at 37°C with stirring for 24 hours. 35 μL of each sample was dispersed in 400 μL of fetal bovine serum. After 30 minutes, both the radiolabeled sample suspension and the serum sample suspension were subjected to instantaneous thin-layer chromatography (iTLC) under the following conditions: a 5 μL drop was placed on a glass fiber iTLC strip, which was then placed in the TLC chamber using 20 mM sodium citrate (pH 6.5) as the mobile phase. Upon completion, the application point and the TLC peak were measured separately using a Hidex gamma counter. This experiment demonstrated radiolabeling with a radiochemical purity (RCP) of 55% and a 48% retention rate of Lu in the MOF in serum.

[0191] [Table 4]

[0192] Table 3 shows the radiolabel (RCP) and serum retention of Lu-177 labeled UiO-66(COOH)2 (UiO-66-Lu) and the control sample LuCl3.

[0193] Example 8: Biodistribution of β-emitter-loaded UIO-66-(COOH)2 in healthy mice The biodistribution of Lu177-loaded UIO-66-(COOH)2 was evaluated in healthy mice. Whole-body PET / CT scans were performed from 0 to 4 days after injection. Mice were sacrificed on day 4 after injection to analyze isotope activity in all major organs.

[0194] procedure: Lu177-loaded anti-EpCAM conjugate UIO-66-(COOH)2 was resuspended in 5% dextrose solution before administration to healthy mice. Free Lu177 dissociated in 5% dextrose served as a control. Mice were scanned by PET / CT on days 0–4 post-administration and finally sacrificed to collect liver, kidney, spleen, femur, skin, blood, small intestine, large intestine, lung, and heart. Activity was measured by gamma-ray emission analysis using HIDEX.

[0195] The radiolabeling dose was 10 MBq in a 200 μL suspension of PEGylated UIO-66-(COOH)2 (0.8 mg) in 0.015 mol / L choline phosphate buffer (pH 6.9) and 5 wt% dextrose. 177 The reaction was carried out by adding 10 MBq of [Lu]LuCl3 solution to 200 μL of 0.015 mol / L choline phosphate buffer (pH 6.9) and 5 wt% dextrose. 177 A control sample consisting of [Lu]LuCl3 was prepared in parallel. The Lu samples were kept at 37°C with stirring for 24 hours. 100 μL of each dose was injected into healthy female nude mice (6 weeks old).

[0196] FIG. 17 shows the biodistribution of free Lu177 and Lu177-labeled anti-EpCAM-conjugated UIO-66-(COOH)2 (NNP-Lu177) in mouse tissues, and the in vivo retention of Lu177 in the MOF scaffold.

[0197] Example 9: Lu177 radiolabeled MOF-anti-EpCAM binds to colorectal cancer cells This experiment demonstrates that Lu177 radiolabeled anti-EpCAM conjugate UIO-66-(COOH)2 binds to EpCAM-positive colon cancer cells (HCT116).

[0198] Procedure: MOF-anti-EpCAM was radiolabeled (same procedure as in Example 7). The complex was incubated with HCT116 cells and 5x10 6 cells / reaction and 1 x 106 The cells / reactions were mixed and incubated for 60 minutes. The supernatant and cell fraction were separated by centrifugation (300 g for 5 minutes) prior to measurement of gamma-ray emission using HIDEX.

[0199] FIG. 18 shows that anti-EpCAM conjugate UIO-66-(COOH) 2 radiolabeled with Lu177 binds to colorectal cancer cells HCT116.

[0200] Example 10: Designed Experiment; In Vivo Safety Threshold of Beta-Emitter-Loaded UIO-66-(COOH)2 with and without Ra223 A maximum tolerated dose (MTD) study will be conducted to assess the maximum safe dose for further in vivo studies. Weight loss and general behavior will be assessed. Mice will be sacrificed if severe symptoms appear.

[0201] procedure: The MTD is defined as the maximum tolerated dose at which survival is possible without serious side effects. Six doses are used, starting at 55 MBq / kg and increasing in two-fold increments up to 1870 MBq / kg per prototype (UIO-66-(COOH)2 anti-HER2, UIO-66-(COOH)2 anti-EpCAM, UIO-66-(COOH)2 anti-MUC5AC, or UIO-66-(COOH)2PEG), with two mice per dose level.

[0202] Example 11: Viability studies of PEG-conjugated UIO-66-(COOH)2 (NPs) showed no toxicity in vitro Exposure of HT29 and HCT116 to increasing doses of NP for 2 days before analyzing cell viability indicates that NP does not induce cell death under these conditions.

[0203] procedure: HT29 or HCT116 cells were exposed to 0.001–500 μg of UIO-66-(COOH)2 (NPs) per well and cultured at 37°C and 5% CO2 in a humidified incubator for 24 and 48 hours. Afterward, the supernatant was discarded, and 100 μL of fresh cell culture medium (10% fetal bovine serum (FBS) from Roswell Park Memorial Institute (RPMI)) and 20 μL of CellTiter 96® AQueous One Solution Reagent (Promega) containing the tetrazolium compound [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt; MTS] and electron coupling reagent (phenazine ethosulfate; PES) were added. Each plate was incubated in a humidified incubator at 37°C and 5% CO2 for 1.5 hours, after which the absorbance at 490 nm was measured using a plate reader.

[0204] See Figures 16A and B), absorbance did not change across all conditions, thus suggesting that cell content per well did not change over the 48 hour period.

[0205] Example 12: Designed Experiment: Biodistribution of UIO-66-(COOH)2 (NPs) conjugated to anti-HER2 antibody, anti-EpCAM antibody, anti-MUC5AC, and polyethylene glycol (PEG) to investigate tumor targeting of NPs with and without antibodies

[0206] procedure: The biodistribution of UIO-66-(COOH)2 targeting EpCAM, HER2, or anti-MUC5AC, along with their tumor localization ability, will be investigated using tumor models representing breast, colorectal, pancreatic, and / or liver cancer. To investigate the specificity of nanoparticles (NPs), NPs with and without antibodies conjugated to the particles in addition to radioactivity will be used. To examine the biodistribution of the drug over time and whether its biodistribution is specific to the tumor, mice will be sacrificed between days 0 and 30 after treatment. Mice will be sacrificed at each time point with each drug. For the two drugs targeting EpCAM, this will be investigated in two or more models (e.g., HCT116 and HT29), using tumors growing in the liver and the peritoneal cavity. For tumors growing in the peritoneal cavity, the agent will be injected either intravenously (i.v.) or intraperitoneally (i.p.) to determine which injection form has the best biodistribution for tumors growing in the peritoneal cavity. Two HER2-targeted agents will be examined in at least two models, such as SKBR3 (cell line) and BBRC160 (PDX), which produce breast, liver, and peritoneal tumors.

[0207] Example 13: Designed Experiment: Efficacy Study of UIO-66-(COOH)2 (NPs) Conjugated to Anti-HER2 Antibody, Anti-EpCAM Antibody, Anti-MUC5AC, and Polyethylene Glycol (PEG) to Examine the Tumoricidal Properties of NPs with and without Antibodies

[0208] procedure: The ability of UIO-66-(COOH)2 carrying the beta-emitters shown in Table 1 to inhibit tumor growth is tested in tumor models representing breast cancer (e.g., SKBR3 and BB6RC160), colorectal cancer (e.g., HCT116, HT29, and SW620), liver cancer models (patient xenograft-derived), and pancreatic cancer models (Panc-1, Capan-1, or Capan-2). Because colorectal, breast, and pancreatic cancers frequently metastasize to the liver and peritoneal cavity, it is appropriate to examine the efficacy of UIO-66-(COOH)2 in these settings, which have a microenvironment more similar to that of tumors in patients.

Claims

1. A particle, a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, said functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs comprising: Optionally, at least one targeting moiety linked to the particle on the outer surface of the particle; and at least one beta-emitting radionuclide; Including, wherein said at least one beta-emitting radionuclide is located in at least one of said pores.

2. 2. The particle of claim 1, wherein the MOF is a Zr(IV), Hf(IV), Ce(IV), Fe(III), Al(III), Ti(IV), or Cr(III)-based MOF with a carboxylate organic monomer, or a Zn(II) imidazolate-based MOF.

3. The MOF is UiO-66-COOH, MOF-808, or UIO-66(COOH). 2 3. The particle according to claim 1 or 2, wherein

4. 4. The particle according to any one of claims 1 to 3, wherein the beta-emitting radionuclide is selected from the group comprising beta-emitting isotopes having beta-excitation energies in the range of 0.3 MeV to 3.5 MeV, and / or the beta-emitting radionuclide has a half-life in the range of 2 hours to 150 days.

5. 5. The particle of any one of claims 1 to 4, wherein the beta-ray emitting radionuclide comprises or is selected from the group consisting of sodium-24, phosphorus-32, potassium-42, calcium-47, scandium-47, iron-59, copper-64, copper-67, strontium-89, yttrium-90, molybdenum-99, ruthenium-103, indium-132, cerium-141, samarium-153, dysprosium-165, holmium-166, erbium-169, thulium-170, lutetium-177, tantalum-182, rhenium-186, rhenium-188, iridium-192, and gold-198.

6. The particle according to any one of claims 1 to 5, wherein the particle is a nanoparticle.

7. A particle according to any one of claims 1 to 6, wherein said at least one targeting moiety is present and is preferably an antibody.

8. 8. The particle of any one of claims 1 to 7, wherein the particle comprises more than one radionuclide, at least one radionuclide being a beta-emitting radionuclide and one or more other radionuclides being selected from the group of alpha-emitting radionuclides.

9. In the first container: A particle, a repeating three-dimensional network of inorganic and organic monomers that forms pores; and at least one free functional group extending into the pore, said functional group being selected from the group including carboxylic acid, carboxylate, hydroxyl, sulfonic acid, sulfonate, sulfhydryl, primary amine, secondary amine, and combinations thereof; MOFs comprising: Optionally, at least one targeting moiety attached to said particle on the outer surface of said particle; and In the second container, beta-emitting radionuclides Kit including:

10. A composition comprising at least one particle according to any one of claims 1 to 8 together with at least one pharmaceutically acceptable carrier, diluent and / or excipient.

11. A particle according to any one of claims 1 to 8 or a composition according to claim 10 for use in a method for the treatment of a proliferative disease.

12. The particles of any one of claims 1 to 8, wherein the proliferative disease is cancer and the treatment is for a solid tumor or a non-vascular tumor, or the composition of claim 10 for use as claimed in claim 11.

13. A particle according to any one of claims 1 to 8 or a composition according to claim 10 for use in a method for the treatment of liver cancer, wherein the particle does not comprise a targeting moiety.

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