Size-controlled radioactive labeled particles

Abstract

This disclosure relates to particles comprising degradable compounds, radionuclides, and phosphorus-containing additives. Phosphorus-containing additives, such as phosphonates, have a unique ability to control the size of medical particles. These applications allow the particles to be used as pharmaceuticals, and particularly for imaging in the field of cancer.

Description

Technical Field

[0001] This disclosure relates to particles comprising degradable compounds, radionuclides, and phosphorus-containing additives. Phosphorus-containing additives, such as phosphonates and phosphates, have a unique ability to control the size of medical particles. These applications allow the particles to be used as pharmaceuticals, and particularly for imaging in the field of cancer. Background Technology

[0002] Radiolabeled microparticles have shown promise in cancer radiotherapy and pain management, such as radioactive synovectomy for arthritis (which involves injecting small radioactive particles into the joint to treat synovitis).

[0003] Slowly degradable nanoparticles and microparticles are preferred, and for this purpose, non-toxic salt crystalline particles are promising. The morphology and size of these crystalline particles may become unstable over time, especially when the particles are in suspension. Importantly, the particles can be radiolabeled in good yield, and the size distribution and morphology of the crystalline particles remain stable over time when injected into the patient.

[0004] In some applications, understanding the biodistribution of particles can be useful before administering a radiotherapy particle suspension. Particles made from stable particles radiolabeled with radionuclides suitable for imaging can be used for this purpose because it allows imaging of particle distribution within the body. Particles with defined size distributions can be used for imaging purposes to assess fluid distribution in different body compartments, and they can also be applied to specific treatments for certain types of cancer and can be designed specifically for certain types of cancer.

[0005] Requirements for such particles used in medical applications involve controlling particle size and designing them in a manner best suited to their intended use. The particles can be non-degradable or degradable. One advantage of using non-degradable particles is the reduced risk of systemic toxicity. Disadvantages include potentially more uneven radiation dose distribution and some localized toxicity risks from “hot spots.” Stable radiotherapy particles have been used for radioembolization by using stable markers on non-degradable glass spheres (TheraSphere). TM ) or resin-based spheres (SIR-Spheres) TM High-energy beta emitter on ) 90 Y is used to treat primary tumors and liver metastases. In this case, the liver tissue will shield the intestines and other organs from toxic radiation.

[0006] Potential advantages of using degradable particles that slowly release some radionuclides include a more uniform radiation dose distribution due to the diffusion of the parent nuclide and / or short-lived daughter nuclides, and the likelihood of "hot spots" causing local toxicity. Potential disadvantages include the possibility of systemic toxicity due to the potential transport of released radionuclides into the bloodstream and further redistribution. Degradable particles are primarily used with other cytotoxic compounds, such as chemotherapeutic agents, and are not currently used with radionuclides.

[0007] A major problem with radiolabeled degradable crystalline particles is the recrystallization and instability of particle size in suspensions over time / during storage, the rate of which can be increased, for example, by energy input, such as autoclaving. Particles change size distribution and morphology over time in various suspensions, and also when autoclaved in all common solutions tested (including saline, PBS, and TRIS).

[0008] Therefore, improved radionuclide delivery systems based on particles made from non-toxic salts are of great significance for medical applications, and further stabilization of degradable particle size patterns is assured. Summary of the Invention

[0009] In a broad sense, this invention relates to particles containing degradable compounds, radionuclides, and phosphorus-containing additives.

[0010] The degradable compound can be selected from CaCO3, MgCO3, SrCO3, BaCO3, calcium phosphate including hydroxyapatite Ca5(PO4)3(OH) and fluorapatite, and complexes with any of these as the main component. The degradable compound is CaCO3.

[0011] Phosphorus-containing additives can be phosphates, which are selected from orthophosphates, linear oligophosphates, polyphosphates, and cyclic polyphosphates.

[0012] Polyphosphates can be selected from pyrophosphates, tripolyphosphates, and triphosphonic phosphates. Phosphorus-containing additives can be cyclic polyphosphates, such as sodium hexametaphosphate (SHMP). Phosphorus-containing additives can be phosphonates. Phosphonates can be bisphosphonates. Bisphosphonates can be selected from etidronate, clodronate, tiludronate, pamidronate, neridronate, opaldronate, alendronate, ibandronate, risedronate, and zoledronic acid. Phosphonates can be polyphosphonates. Polyphosphonates can be selected from EDTMP-ethylenediaminetetra(methylenephosphonic acid), DOTMP-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacyl-tetra(methylphosphonic acid), and DTPMP-diethylenetriaminepenta(methylenephosphonic acid).

[0013] Radionuclides can be selected from225 Ra、 224 Ra、 223 Ra、 225 Ac、 227 Th、 211 At、 213 Bi、 64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y、 220 Rn、 216 Po、 212 Pb, 212 Bi、 212 Po、 208 Tl、 18 F, 67 Ga、 99m Tc, 111 In、 203 Pb, 64 Cu、 152 Tb and 155 Tb. The radionuclide can be selected from alpha-radioactive nuclides suitable for treatment, which are derived from... 225 Ac、 211 At、 213 Bi、 212 Bi、 225 Ra、 224 Ra、 223 Ra and 227 Th is the composition. Radionuclides can be selected from suitable β-radionuclides for treatment, which are composed of... 64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y composition. Radionuclides can be β emitters with α-generations suitable for treatment, i.e. 212 Pb. Radionuclides can be selected from those emitting alpha. 224 Ra, which has daughter radionuclides 220 Rn、 216 Po、 212 Pb, 212Bi、 212 Po and 208 Tl. Radionuclides can be selected from those suitable for imaging, which are derived from... 18 F, 67 Ga、 99m Tc, 111 In、 203 Pb, 64 Cu、 152 Tb and 155 Tb composition.

[0014] The particle size can range from 1 nm to 500 μm.

[0015] The degradable compounds can be selected from PEG-modified CaCO3, protein-modified CaCO3 including mAb and Fab, carbohydrate-modified CaCO3, lipid-modified CaCO3, vitamin-modified CaCO3, organic compound-modified CaCO3, polymer-modified CaCO3 and / or inorganic crystal-modified CaCO3.

[0016] One aspect of the present invention relates to compositions comprising one or more particles of the present invention.

[0017] The composition may be a pharmaceutical composition comprising one or more particles of the present invention, as well as a diluent, a carrier, a surfactant, and / or an excipient.

[0018] The compositions of the present invention can be prepared with a dose of radionuclides ranging from 1 kBq to 10 GBq per dose or with a dose of radionuclides ranging from 50 MBq to 100 GBq suitable for multi-dose industrial-scale production.

[0019] The composition can be a particle suspension containing monodisperse or polydisperse particles of the present invention.

[0020] The compositions of the present invention are suitable for parenteral use, such as intravenous, intracavitary and / or intratumoral injection.

[0021] One aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention used as pharmaceuticals.

[0022] One aspect of the present invention relates to the use of the particles, compositions or pharmaceutical compositions of the present invention for endovascular treatment, radioembolization or radiotherapy for synovectomy.

[0023] One aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention for the treatment of cancer.

[0024] One aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention for treating cancer, wherein the cancer is selected from intraperitoneal carcinoma, intracranial carcinoma, pleural carcinoma, bladder cancer, gastric cardia cancer, subarachnoid carcinoma, non-cavitary targets such as melanoma, and non-small cell lung cancer.

[0025] One aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention for imaging.

[0026] One aspect of the invention relates to the particles, compositions, or pharmaceutical compositions of the invention used according to the invention in combination with other cancer therapies, such as chemotherapy such as taxanes (e.g., paclitaxel, docetaxel), platinum-based therapies (e.g., carboplatin, cisplatin, doxorubicin, mitomycin), DNA repair inhibitors such as PARP inhibitors (e.g., olaparib, rucaparib, niraparib, talaparib, veliparib, pamiparib, CEP 9722, E7016, and 3-aminobenzamide), and radioimmunotherapy.

[0027] The concentration of phosphonates and / or phosphate compounds is from 1 microgram to 1,000 milligrams per milliliter, for example, from 0.1 milligrams to 10 milligrams per milliliter of final solution, or from 1 microgram to 1,000 milligrams per gram of particles in the final solution.

[0028] One aspect of the invention relates to a method for preparing particles according to the invention, the method comprising contacting a degradable compound, a radionuclide, and a phosphorus-containing additive with or without a radionuclide carrier. The degradable compound and the radionuclide may form particles in an initial step, followed by control of their size using the phosphorus-containing additive. Detailed Implementation

[0029] The inventors were surprised to discover that applying phosphorus-containing additives can enhance several beneficial effects in the preparation and maintenance of particles made from biodegradable compounds and containing radioactive nuclides. Such compounds are typically non-toxic salts, such as crystalline particles of non-toxic salts. These particles can be used for a variety of medical applications, including cancer treatment and imaging.

[0030] Some beneficial effects include stability over time and at higher temperatures (e.g., during autoclaving). Enhanced stability maintains size distribution, morphology, and / or does not lead to the release of particle-associated radionuclides. These parameters are critical in the production, transport, and storage of products for clinical use. It is also important to recognize that particles can be designed according to their intended use. Furthermore, large particles can be designed for, for example, radioembolization, in which large particles can become trapped in blood vessels surrounding a tumor, while microparticles can be suitable for cavitary therapies where it is ideal to prevent rapid passage of particles through membranes (e.g., the peritoneum), whereas nanoparticles are used for intravenous administration. Phosphorus-containing additives, such as bisphosphonates, polyphosphonates, and polyphosphates, have been found to act as particle stabilizers, thereby maintaining size distribution and preventing the release of particle-associated radionuclides. After autoclaving at 120°C and storage at room temperature for several days, the microparticles retain their size and radiolabeling, enabling aseptic production, allowing time for transport, and time for processing the radiolabeled particles for clinical use (see, for example, Examples 6, 7, and 11).

[0031] Therefore, the particles and compositions of the present invention have the beneficial technical advantages of enhanced stability and prevention of particle recrystallization over time in suspensions and / or during storage. The rate of recrystallization and size change can be increased, for example, by energy input, such as by autoclaving. Particles change their size distribution and morphology over time in various suspensions, and also when autoclaved in all common solutions tested (including saline, PBS, TRIS). Therefore, phosphorus-containing additives can be used to increase the stability of particles in compositions or suspensions. Phosphorus-containing additives can thus be used to prevent particle recrystallization, for example, recrystallization that occurs over time and / or with increased energy input, such as autoclaving.

[0032] Therefore, the present invention relates to particles comprising a degradable compound, a radionuclide, and a phosphorus-containing additive. The phosphorus-containing additive may be incorporated into the particles, associated with the particle surface, or present around the particles, i.e., present in the composition or suspension to which the particles belong. Therefore, one aspect of the invention relates to a composition or suspension comprising particles, wherein the particles comprise a degradable compound, a radionuclide, and a phosphorus-containing additive, and wherein the phosphorus-containing additive is associated with the particles by being present in the composition or suspension. It may be part of the particles. It may be located on the surface of the particles. It may be located in a dispersion of the particles. It may be part of the composition or suspension and / or dispersion of the particles. It may also be part of the particles and part of the composition or suspension of the particles. These individual components may be combined to form different types of particles, which have different properties depending on the intended use of the particles.

[0033] This means that phosphorus-containing additives, such as EDTMP and / or pamidronate, are typically present, at least in trace amounts, on or bound to individual particles. Therefore, if the phosphorus-containing additive content of particles in a composition (e.g., a solution) is tested, the phosphorus-containing additive, such as EDTMP and / or pamidronate, is typically found, at least partially, on or within the particles. The total amount of phosphorus-containing additives (such as EDTMP and / or pamidronate) in the composition (e.g., a solution) will vary depending on the particle design, but generally means that at least 0.01–80% of the total phosphorus-containing additive in the composition will be in or on the particles, with the remainder in the composition. The at least 0.01–80% of the total phosphorus-containing additive in or on the particles can be 0.1–50%, for example 10–50%, for example 20–80%, for example 10–80%.

[0034] Degradable compounds

[0035] The degradable compounds of this invention can be any degradable compound. Degradation can be accomplished through any pathway selected from high pH, ​​low pH, temperature, proteases, enzymes, nucleases, and / or through cellular processes such as endocytosis, where endocytosis also includes phagocytosis. Therefore, the degradable compounds can be non-toxic salts or non-toxic salt crystals.

[0036] In one or more embodiments of the invention, the degradable compound may be selected from CaCO3, MgCO3, SrCO3, BaCO3, calcium phosphate including hydroxyapatite Ca5(PO4)3(OH) and fluorapatite, and complexes with any of these as the main component. The main component is defined as at least 20% of the total molecular weight of the particles, for example, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99%.

[0037] The degradable compound can be CaCO3, selected from PEG-modified CaCO3, protein-modified CaCO3 including mAb and Fab, carbohydrate-modified CaCO3, lipid-modified CaCO3, vitamin-modified CaCO3, organic compound-modified CaCO3, polymer-modified CaCO3, and / or inorganic crystal-modified CaCO3.

[0038] The degradable compound can be MgCO3, selected from PEG-modified MgCO3, protein-modified MgCO3 including mAb and Fab, carbohydrate-modified MgCO3, lipid-modified MgCO3, vitamin-modified MgCO3, organic compound-modified MgCO3, polymer-modified MgCO3, and / or inorganic crystal-modified MgCO3.

[0039] The degradable compound can be SrCO3, selected from PEG-modified SrCO3, protein-modified SrCO3, including mAb and Fab, carbohydrate-modified SrCO3, lipid-modified SrCO3, vitamin-modified SrCO3, organic compound-modified SrCO3, polymer-modified SrCO3 and / or inorganic crystal-modified SrCO3.

[0040] The degradable compound can be BaCO3, selected from PEG-modified BaCO3, protein-modified BaCO3 including mAb and Fab, carbohydrate-modified BaCO3, lipid-modified BaCO3, vitamin-modified BaCO3, organic compound-modified BaCO3, polymer-modified BaCO3 and / or inorganic crystal-modified BaCO3.

[0041] The degradable compound can be Ca5(PO4)3(OH), selected from PEG-modified Ca5(PO4)3(OH), protein-modified Ca5(PO4)3(OH), including mAb and Fab, carbohydrate-modified Ca5(PO4)3(OH), lipid-modified Ca5(PO4)3(OH), vitamin-modified Ca5(PO4)3(OH), organic compound-modified Ca5(PO4)3(OH), polymer-modified Ca5(PO4)3(OH), and / or inorganic crystal-modified Ca5(PO4)3(OH).

[0042] The degradable compound can be fluorapatite, selected from PEG-modified fluorapatite, protein-modified fluorapatite including mAb and Fab, carbohydrate-modified fluorapatite, lipid-modified fluorapatite, vitamin-modified fluorapatite, organic compound-modified fluorapatite, polymer-modified fluorapatite, and / or inorganic crystal-modified fluorapatite.

[0043] Composite particles may contain two or more of these degradable compounds, which combine to form the main components, as defined above.

[0044] Degradable compounds can be used as complexes with other salts or proteins or peptides and can be surface-modified by surfactants such as oleates.

[0045] In one specific embodiment, the degradable compound is used in conjunction with a compound selected from polyethylene glycol (PEG) modified degradable compound particles or inorganic crystal modified degradable compounds.

[0046] In one specific embodiment, the degradable compound is modified with functional receptor and / or antigen-binding groups, including monoclonal antibodies and derivatives, as well as vitamins and derivatives, allowing the receptor or antigen particles to bind to individual target cells and diseased tissues. This means that particle modification involves adding other compounds to the degradable compound. This can be accomplished in a variety of ways, such as through dipole-dipole interactions, ion-dipole and ion-induced dipole forces, hydrogen bonds, van der Waals forces, and the relative strength of the forces.

[0047] Chelating agents can be used, which preferentially conjugate to target affinity molecules, such as monoclonal or polyclonal antibodies or antibody derivatives, vitamins or vitamin derivatives.

[0048] Monoclonal antibodies (mAb), polyclonal antibodies (pAb), antigen-binding fragments (Fab), and other types of peptides and proteins can be used to contain specific targets in particles, meaning that by adding specific target molecules, the particles will be able to enhance their affinity for certain target cells in the body.

[0049] Phosphorus additives

[0050] Phosphorus-containing additives can be phosphates, thus becoming phosphate-containing additives. Phosphorus-containing additives can also be phosphonates, thus becoming phosphonate-containing additives.

[0051] Phosphonates and phosphonic acids are organophosphorus compounds containing C-PO(OH)2 or C-PO(OR)2 groups (where R = alkyl or aryl). Phosphonic acids, usually treated as salts, are generally non-volatile solids that are poorly soluble in organic solvents but soluble in water and common alcohols. Therefore, various salts and acids of phosphonates are also considered part of the definition of phosphonates.

[0052] Phosphoric acid in the general sense is an oxyphosphate, in which each phosphorus atom is in the +5 oxidation state and is bonded to four oxygen atoms, one of which is arranged in a tetrahedral corner through a double bond. (The text then abruptly shifts to a seemingly unrelated topic: removing the H atom as a proton.) + The hydrogen atoms convert phosphoric acid into phosphate anions. Partial removal produces various hydrogen phosphate anions.

[0053] Phosphorus-containing additives can be phosphonates. Phosphonates can be bisphosphonates. Bisphosphonates can be selected from etidronate, clodronate, tiludronate, pamidronate, neridronate, opaldronate, alendronate, ibandronate, risedronate, and zoledronic acid. In one or more embodiments of the present invention, the bisphosphonate is etidronate. In one or more embodiments of the present invention, the bisphosphonate is clodronate. In one or more embodiments of the present invention, the bisphosphonate is tiludronate. In one or more embodiments of the present invention, the bisphosphonate is pamidronate. In one or more embodiments of the present invention, the bisphosphonate is neridronate. In one or more embodiments of the present invention, the bisphosphonate is opaldronate. In one or more embodiments of the present invention, the bisphosphonate is alendronate. In one or more embodiments of the present invention, the bisphosphonate is ibandronate. In one or more embodiments of the present invention, the bisphosphonate is risedronate. In one or more embodiments of the present invention, the bisphosphonate is zoledronic acid.

[0054] Phosphonates can be polyphosphonates. Polyphosphonates can be selected from EDTMP-ethylenediaminetetra(methylenephosphonic acid), DOTMP-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacyl-tetra(methylphosphonic acid), and DTPMP-diethylenetriaminepenta(methylenephosphonic acid). In one or more embodiments of the present invention, the phosphonate is EDTMP-ethylenediaminetetra(methylenephosphonic acid). In one or more embodiments of the present invention, the phosphonate is DOTMP-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacyl-tetra(methylphosphonic acid). In one or more embodiments of the present invention, the phosphonate is DTPMP-diethylenetriaminepenta(methylenephosphonic acid).

[0055] In one embodiment, the particles of the present invention comprise CaCO3 as a degradable compound, EDTMP-ethylenediaminetetra(methylenephosphonic acid) as a phosphorus-containing additive, and a radionuclide. 224 Ra.

[0056] In one embodiment, the particles of the present invention comprise CaCO3 as a degradable compound, pamidronate as a phosphorus-containing additive, and a radionuclide. 212 Pb.

[0057] Phosphate-containing additives can be selected from orthophosphates, linear oligophosphates, polyphosphates, and cyclic polyphosphates. Polyphosphates can be selected from pyrophosphates, tripolyphosphates, and triphosphonic phosphates. Phosphate-containing additives can be cyclic polyphosphates, such as sodium hexametaphosphate (SHMP).

[0058] The concentration of phosphonates and / or phosphate compounds ranges from 1 μg to 1000 mg per mL, for example, 0.1 mg to 10 mg per mL of final solution, or 1 μg to 1000 mg per gram of particles in the final solution. Using the particles and methods used in Example 6, where the range of 0.001–0.046 g EDTM per gram of CaCO3 was tested, at least 0.013 g EDTMP per gram of CaCO3 particles was required for size control. For SHMP and pamidronate, all tested concentrations of 0.015–0.200 g SHMP / per gram of CaCO3 and 0.008–0.016 g pamidronate / per gram of CaCO3 resulted in size stability of the polydisperse suspension. Example 12 ( Figure 5 This indicates that adding at least 1% EDTMP can control the size of the particles.

[0059] Therefore, for phosphate-containing additives, such as EDTMP, the patient dosage range can be from 1 microgram to 1 gram per milliliter or from 1 microgram to 1 gram per gram of particles. For example, the patient dosage can be 2.5 mg / mL or 25 mg / mL of a degradable compound, such as CaCO3. The amount of phosphate-containing additives, such as EDTMP, added can be 0.1-10%. The addition amount can also be 0.5-5% or 0.5-2%. The addition amount can be at least 1%.

[0060] For 10 ml of product, the concentration of phosphate-containing additives (such as EDTMP) can be 24.4 mg, for example, a range of 20 to 30 mg per 10 ml of product.

[0061] Phosphate-containing additives, such as EDTMP, can be used as excipients at a final concentration of 5.6 mM (2.44 mg / mL), ranging from 4.6 to 6.6 mM.

[0062] Phosphate-containing additives, such as EDTMP, can be used in the range of 24.4 mg per gram of calcium carbonate or 10 mg–50 mg per gram.

[0063] Phosphate-containing additives, such as EDTMP, can be used at a concentration of 2.4% w / w, ranging from 1 to 20% w / w.

[0064] Phosphate-containing additives, such as EDTMP, can be used at a dose of 24.4 mg per 10 ml of product.

[0065] Phosphate-containing additives, such as EDTMP or EDTMPA (acid), can be used as excipients at a concentration of 5.6 mM (2.44 mg / mL).

[0066] The concentration of phosphate-containing additives, such as EDTMP or EDTMPA (acid), can be 5.6 mM ± 15%.

[0067] For additives containing phosphonates, such as pamidronate, the patient dosage range can be 1 μg to 1 g per milliliter or 1 μg to 1 g per gram of particles. For example, the patient dosage can be 2.5 mg / mL or 25 mg / g of a degradable compound, such as CaCO3. The amount of phosphate-containing additives, such as pamidronate, can be 0.1-10%. The addition amount can also be 0.5-5% or 0.5-2%. The addition amount can be at least 1%.

[0068] For 10 ml of product, phosphate-containing additives, such as pamidronate, can be used at a concentration of 10 mg, for example, 5 to 50 mg per 10 ml of product.

[0069] Phosphate-containing additives, such as pamidronate, can be used as excipients at a final concentration of 4 mM (1 mg / mL), ranging from 0.1 to 10 mM.

[0070] Phosphate-containing additives, such as pamidronate, can be used in the range of 0.01 g / g calcium carbonate or 1 mg to 50 mg / g calcium carbonate.

[0071] Phosphate-containing additives, such as pamidronate, can be used at a concentration of 1% w / w, ranging from 0.1% to 5% w / w.

[0072] Phosphate-containing additives, such as pamidronate, can be used in 10ml of product at a dosage of 10mg.

[0073] Phosphate-containing additives, such as pamidronate or pamidronic acid, can be used as excipients at a concentration of 4 mM (1 mg / mL).

[0074] Phosphate-containing additives, such as pamidronate or pamidronic acid, can be present at a concentration of 4 mM ± 25%.

[0075] Example 13 shows that the sedimentation rate decreases with decreasing particle size. Several improvements in suspension treatment offer advantages for clinical product administration. Therefore, the addition of phosphate-containing compounds (such as EDTMP) can significantly influence sedimentation rate by reducing particle size. Example 14 shows... 212 Pb and 224 A comparison of Ra retention on CaCO3 particles with and without a layer of encapsulation shows how adding this layer can improve, particularly... 212 Pb retention. Example 15 shows a comparison of the biodistribution of radium-224 and lead-212 between different amounts of layered encapsulated microparticles and free radium-224. Bone resorption levels decreased with increasing microparticle amounts. Example 16 shows a chemically equivalent stable daughter nuclide adsorbed on MP. 208The cumulative amount of Pb increases over time. Example 17 shows EDTMP and... 212 The known complexing properties of Pb and calcium indicate that 212 The Pb-EDTMP complex may also associate with MP.

[0076] Radionuclides

[0077] Therapeutic applications are considered for radionuclides exhibiting α, β, and Auger electron emission. Due to variations in the mass and energy of the emitted particles, their extent within tissues and their linear energy transfer (LET) (defined as the energy transferred to matter per unit length) differ significantly. Generally, low-LET β-emitters are considered more suitable for treating larger tumors than high-LET α and Auger emitters, while high-LET α and Auger emitters are preferred for treating micrometastatic and single-cell diseases. Therefore, the radionuclides in the particles of this invention can be tailored to the intended use.

[0078] The main medical advantage of compounds that emit alpha particles in local treatments, such as intraperitoneal therapy, is their shorter range; alpha particles typically have a range of less than 0.1 mm, while the range of beta particles in medical beta emitters is from millimeters to centimeters.

[0079] In intraperitoneal (ip) use, the use of alpha emitters in an intracavitary environment reduces the risk of toxicity from irradiating deeper areas of the viscera, such as radiation-sensitive intestinal crypt cells. The high linear energy transfer of the emitted alpha particles is also advantageous because few alpha hits are required to kill cells and cellular resistance mechanisms, such as the high repair capacity for DNA strand breaks, is not a significant problem due to the high likelihood of generating irreparable double-strand breaks.

[0080] The high efficiency of each decay means less radioactivity is needed, reducing the need for shielding hospital staff and their families, since most alpha and beta emitters also emit some X-rays and gamma rays that require shielding.

[0081] In cases where cancer is characterized as a large-volume disease, beta particles, with their longer range compared to alpha particles, may be advantageous. The longer path length of beta particles leads to the so-called crossfire effect, where significantly higher radiation is emitted from adjacent and distant cells, thus damaging cells farther from the radiolabeled particles.

[0082] In this paper, progeny are understood as radionuclides, which are the result of the decay of the parent radionuclide. Therefore, when 224 When Ra is a parent radionuclide, it produces 220 Rn (daughter radionuclides) 216 Po (a radioactive nuclide) and 212 Pb (a radionuclide found in the great-grandson body). Therefore, it is believed that...220 Rn 、216 Po and 212 Pb is 224 Progeny radionuclides of Ra.

[0083] Therefore, in one implementation, it is a radionuclide that emits alpha. 224 Ra and daughter radionuclides 220 Rn, radioactive nuclide 216 Po and great-grandson radionuclides 212 Pb. For the particles of this invention, when 224 When Ra is a radioactive nuclide, all of these radioactive nuclides will be contained within the particle.

[0084] The radionuclides in the particles of this invention can be selected from... 225 Ra、 224 Ra、 223 Ra、 225 Ac、 212 Bi、 227 Th、 211 At、 213 Bi、 212 Pb, 64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y、 220 Rn、 216 Po、 212 Po、 208 Tl、 18 F, 67 Ga、 86 Y、 99m Tc, 111 In、 203 Pb, 83 Sr、 152 Tb and 155 Tb. The radionuclide can be selected from alpha-radioactive nuclides suitable for treatment, which are derived from... 225 Ac、 211 At、 213 Bi、 212 Bi、 225 Ra、 224 Ra、 223 Ra and 227 Th is the composition. Radionuclides can be selected from suitable β-radionuclides for treatment, which are composed of...64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y composition. Radionuclides can be β emitters with α-progeny suitable for treatment, i.e., progeny... 212 Pb. Radionuclides can be selected from alpha emission. 224 Ra and progeny radionuclides 220 Rn、 216 Po、 212 Pb, 212 Bi、 212 Po and 208 Tl. Radionuclides can be selected from those suitable for imaging, which are derived from... 18 F, 67 Ga、 86 Y、 99m Tc, 111 In、 203 Pb, 64 Cu、 83 Sr、 152 Tb and 155 Tb composition. In one or more embodiments of the present invention, the radionuclide is... 225 Ra. In one or more embodiments of the invention, the radionuclide is 224 Ra. In one or more embodiments of the invention, the radionuclide is 223 Ra. In one or more embodiments of the invention, the radionuclide is 225 Ac. In one or more embodiments of the present invention, the radionuclide is 212 Bi. In one or more embodiments of the present invention, the radionuclide is 227 Th. In one or more embodiments of the present invention, the radionuclide is 211 At. In one or more embodiments of the invention, the radionuclide is 213 Bi. In one or more embodiments of the present invention, the radionuclide is 212 Pb. In one or more embodiments of the present invention, the radionuclide is 64 Cu. In one or more embodiments of the invention, the radionuclide is 67 Cu. In one or more embodiments of the invention, the radionuclide is 166Ho. In one or more embodiments of the invention, the radionuclide formula 177 Lu. In one or more embodiments of the present invention, the radionuclide is 32 P. In one or more embodiments of the invention, the radionuclide formula 186 Re. In one or more embodiments of the invention, the radionuclide is 153 Sm. In one or more embodiments of the present invention, the radionuclide is 89 Sr. In one or more embodiments of the invention, the radionuclide is 161 Tb. In one or more embodiments of the invention, the radionuclide is 90 Y. In one or more embodiments of the present invention, the radionuclide is 220 Rn. In one or more embodiments of the present invention, the radionuclide is 216 Po. In one or more embodiments of the invention, the radionuclide is 212 Bi. In one or more embodiments of the present invention, the radionuclide is 212 Po. In one or more embodiments of the invention, the radionuclide is 208 Tl. In one or more embodiments of the present invention, the radionuclide is 18 F. In one or more embodiments of the invention, the radionuclide is 67 Ga. In one or more embodiments of the present invention, the radionuclide is 99m Tc. In one or more embodiments of the present invention, the radionuclide is 111 In one or more embodiments of the present invention, the radionuclide is... 203 Pb. In one or more embodiments of the present invention, the radionuclide is 152 Tb. In one or more embodiments of the invention, the radionuclide is 155 Tb. In one or more embodiments of the invention, the radionuclide is 83 Sr. In one or more embodiments of the present invention, the radionuclide is 86 Y.

[0085] These radionuclides can be incorporated into the particles of the present invention, thus the particles contain one, two, or more of the aforementioned radionuclides. This may occur naturally due to the decay of the radionuclides and their becoming their natural progeny. For example, when 224 Ra is a parent radionuclide, produced by... 220 Rn (daughter radionuclides) 216 Po (a radioactive nuclide) and 212This occurs when Pb (a radioactive isotope) is present. Therefore, 220 Rn、 216 Po and 212 Pb is considered to be 224 The progeny radionuclides of Ra, and due to 224 Ra decays naturally, and they automatically exist in particles in a certain amount.

[0086] The particle may also contain two or more radioactive nuclides, as a higher quantity than natural decay may be beneficial for the particle's intended use. For example, if mixed... 224 Ra and 212 This occurs when Pb forms particles. In this case, compared to using purified Pb... 224 Compared to Ra-prepared particles, the particles contain 212 The Pb content is higher.

[0087] The amount of radionuclide used per patient dose can range from 1 kBq to 10 GBq, more preferably from 100 kBq to 100 MBq, and even more preferably from 0.5 MBq to 25 MBq. A range dose can be from 10 MBq to 10 GBq per patient. A range dose can be from 10 MBq to 5 GBq per patient. This range can be used for beta emitters, alpha emitters, or combinations thereof. The range can be used for treatment or imaging. The dose will be determined based on the type of cancer, such as the aggressiveness of the disease. In one embodiment, the dose is 10-100 kBq / kg, for example, 20-50 kBq / kg. In another embodiment, the dose is 10-1000 kBq / kg, for example, 25-300 kBq / kg. In yet another embodiment, the dose is 100-500 kBq / kg, for example, 150-300 kBq / kg. In one embodiment, the dose is 1-100 MBq / kg, for example, 5-20 MBq / kg. In another embodiment, the dosage is 1-1000 MBq / kg, for example 10-50 MBq / kg. In another embodiment, the dosage is 100-500 MBq / kg, for example 150-300 MBq / kg.

[0088] In one embodiment of the invention, the pharmaceutical composition is prepared with a radionuclide amount of 1 kBq to 10 GBq per dose. For example, if 100 patient doses are produced in a batch per day, this may consist of a total of 1-10 GBq, divided into 100 single-dose vials or ready-to-use syringes.

[0089] In another embodiment of the invention, the pharmaceutical composition is prepared with a radionuclide amount suitable for multi-dose industrial-scale production, for example, 50 MBq to 100 GBq.

[0090] Therefore, the compositions of the present invention can be prepared with a radionuclide amount of 1 kBq to 10 GBq per dose or with a radionuclide amount of 50 MBq to 100 GBq suitable for multi-dose industrial-scale production.

[0091] Form and Composition

[0092] Particles can have a variety of properties, and their size can vary depending on the intended use and application.

[0093] The crystal type can be any known form of degradable compound, and sizes ranging from 1 nm to 500 μm can be used. Sizes can range from 100 nm to 50 μm, more preferably from 1 to 10 μm. In one preferred embodiment, the size is 1 to 10 μm. In another preferred embodiment, the size is 100 nm to 5 μm, and in yet another preferred embodiment, the size is 10 to 100 nm. In yet another preferred embodiment, the size is 1 to 20 μm, and in yet another preferred embodiment, the size is 2 to 10 μm.

[0094] One aspect relates to compositions comprising one or more particles according to the invention. The composition may be a particle suspension comprising monodisperse or polydisperse particles containing degradable compounds, radionuclides, and phosphorus additives.

[0095] One or more embodiments of the present invention relate to the use of the particles of the present invention, wherein the radionuclide is a surface labeled with a radionuclide, an inclusion labeled as part of the particle volume, or a labeled particle on a surface whose radionuclide label is covered by a layer of material to protect the radionuclide-labeled surface and prevent the release of the radionuclide. The particles of the present invention can then be converted into radionuclide-labeled particles, thereby adding a layer of material to cover the original surface to encapsulate the radionuclide. Example 5 describes the preparation of these different types of particles.

[0096] Surface labeling can occur through the adsorption of radionuclides onto crystal particles driven by elemental affinity, or it can be a co-precipitation process where the addition of inorganic compounds facilitates the precipitation. Chelating agents can be used in this process, either incorporated into the particles or on their surface.

[0097] One aspect of the invention relates to a composition comprising particles, said particles comprising a degradable compound and a radionuclide, wherein a phosphorus-containing additive is included in the composition. The composition may be a suspension of the particles. The phosphorus-containing additive may be incorporated into the particles. The phosphorus-containing additive may associate with the particle surface or be present around the particles, i.e., present in the composition or suspension to which the particles belong. Therefore, one aspect of the invention relates to a composition or suspension comprising particles, wherein the particles comprise a degradable compound, a radionuclide, and a phosphorus-containing additive, and wherein the phosphorus-containing additive is associated with the particles by being present in the composition or suspension. The phosphorus-containing additive may be part of the particles. It may be on the surface of the particles. It may be part of the particle composition or suspension. It may also be part of both the particles and the particle composition or suspension.

[0098] One or more embodiments of the present invention relate to a particle suspension, which is a mixture of a solid phase and a liquid phase. The phosphorus-containing additive may be in the liquid phase. The phosphorus-containing additive may be in the solid phase. The phosphorus-containing additive may be in both solid and liquid phases. In the solid phase, the phosphorus-containing additive may be on the surface or embedded in the particles, or both on the surface or embedded in the solid phase. The solid phase may be made of nanoparticles, microparticles, or a combination of both. Radionuclides may be bound to the surface of the particles and / or embedded in the volume or bulk of the particles. Thus, the solid phase may contain particles containing degradable compounds and radionuclides, with or without phosphorus-containing additives, but if the phosphorus-containing additive is not part of the solid phase, it is always a liquid phase. The degradable compounds, radionuclides, and phosphorus-containing additives may be any of those substances disclosed herein.

[0099] Phosphorus-containing compounds may or may not be combined with radionuclides.

[0100] The compositions of the present invention are preferably aqueous compositions. Therefore, in this embodiment, the liquid phase is an aqueous phase. The composition can be a brine composition. The composition can also be an alcohol composition. The composition can be a gel matrix composition. The compositions of the present invention can be suspensions of the particles of the present invention.

[0101] Therefore, another aspect of the present invention relates to compositions or pharmaceutical compositions comprising one or more particles and diluents, carriers, surfactants, antiflocculators and / or excipients according to the present invention.

[0102] Acceptable carriers and drug carriers include, but are not limited to, non-toxic buffers, fillers, isotonic solutions, solvents and co-solvents, antimicrobial preservatives, antioxidants, wetting agents, defoamers, and thickeners. More specifically, drug carriers can be, but are not limited to, physiological saline (0.9%), semi-physiological saline, lactated Ringer's solution, dissolved sucrose, and glucose, such as 3.3% glucose / 0.3% saline. Physiologically acceptable carriers may contain radiation decomposition stabilizers, such as ascorbic acid and human serum albumin, which protect the integrity of the radiopharmaceutical during storage and transportation.

[0103] The particles can be dispersed in a variety of buffer solutions compatible with medical injectables, such as dissolved salts and / or proteins and / or lipids and / or sugars.

[0104] Pharmaceutical compositions may contain a large number of particles. These particles may be the same or different.

[0105] Medical applications

[0106] The particles and compositions of the present invention can be used as radiotherapy compounds and / or radiotherapy mixtures (compositions and solutions).

[0107] One aspect of the invention relates to the particles, compositions, or pharmaceutical compositions of the invention used as medicines. Another aspect of the invention relates to the particles, compositions, or pharmaceutical compositions of the invention for treating cancer.

[0108] Parenteral administration is a term that includes at least intravenous (IV), intramuscular (IM), subcutaneous (SC), and intradermal (ID) administration. Therefore, one or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including parenteral administration. One or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including intravenous (IV) administration. One or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including intramuscular (IM) administration. One or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including subcutaneous (SC) administration. One or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including intradermal (ID) administration. One or more embodiments of the present invention relate to the use of the particles, compositions, or pharmaceutical compositions of the present invention in administration including intratumoral administration.

[0109] The medical uses of the particles of the present invention include human or veterinary uses in the following aspects: (1) endovascular treatment (2) radioembolization (3) radioactive synovectomy (4) imaging (5) as a medical device.

[0110] Endocavitary therapy may include treating cancers such as intraperitoneal cancer, intracranial cancer, pleural cancer, bladder cancer, gastric cardia cancer, and subarachnoid cancer. Examples of cavities in which particles can be used are the cranial cavity, pleural cavity, lung cavity, spinal cavity, pelvic cavity, pericardium, pleural cavity, bladder cavity, or combinations thereof, including cancers that have spread in the peritoneum or meninges and organs within any of these cavities. In one embodiment of the invention, the cancer is selected from intraperitoneal cancer, intracranial cancer, pleural cancer, bladder cancer, gastric cardia cancer, and subarachnoid cancer. In one embodiment of the invention, the cancer is selected from metastatic cancer, lung cancer, ovarian cancer, colorectal cancer, gastric cancer, pancreatic cancer, breast cancer, neoplastic meningitis, peritoneal cancer, pleural effusion, malignant mesothelioma, breast cancer, sarcoma, glioblastoma, and astrocytoma, as well as brain cancer, prostate cancer, bladder cancer, and liver cancer. One or more embodiments of the invention relate to the use according to the invention, wherein the cancer is metastatic cancer. One or more embodiments of the invention relate to the use according to the invention, wherein the cancer is lung cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is ovarian cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is colorectal cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is gastric cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is pancreatic cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is breast cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is neoplastic meningitis. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is peritoneal cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is pleural effusion. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is pleural effusion. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is malignant mesothelioma. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is breast cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is sarcoma. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is brain cancer, such as glioblastoma and astrocytoma. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is bladder cancer. One or more embodiments of the present invention relate to the use according to the invention, wherein the cancer is liver cancer.

[0111] One aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention for treating cancer, wherein the cancer is selected from intraperitoneal carcinoma, intracranial carcinoma, pleural carcinoma, bladder cancer, gastric cardia cancer, subarachnoid carcinoma, non-cavitary targets such as melanoma, and non-small cell lung cancer.

[0112] In one specific embodiment of the use of the particles of the present invention, an infection or inflammation, but not cancer or a disease combined with cancer, is treated or improved. Inflammation can be, for example, arthritis.

[0113] In one embodiment of the invention, the infection is selected from bacterial infection and viral infection.

[0114] Radioembolization can include treating primary or metastatic cancer in an organ, such as the liver, by administering the particles of the present invention to blood vessels leading to a tumor in the liver or another solid organ infiltrated by tumor tissue.

[0115] Radiation-assisted synovectomy for joint diseases, including chronic inflammation, involves targeted radiation therapy using radioactive materials to treat painful joint conditions. Its applications include the treatment of hemophilic arthritis.

[0116] Today, it is based on compounds that emit beta particles for use in inflammatory or rheumatoid arthritis or synovial arthritis of various joints (particularly the knee, hand, and ankle). The degradable particles described in this article may be very useful in radiation synovectomy.

[0117] The particles are preferably administered via local injection, such as intracavitary injection. In one specific embodiment, the particles are injected directly into the tumor.

[0118] Another aspect of the invention relates to a method of treatment or improvement, including applying the particles or pharmaceutical compositions of the invention to an individual in need.

[0119] The compositions of the present invention are suitable for parenteral use, such as intravenous, intracavitary and / or intratumoral injection.

[0120] In one aspect of the invention, the particle according to the invention is a medical device or is contained in a medical device.

[0121] Medical devices are any instruments, equipment, appliances, software, materials, or other articles, whether used alone or in combination, including software that the manufacturer intends specifically for diagnostic and / or therapeutic purposes and which is necessary for its proper application, and software that the manufacturer intends for use in humans to achieve: the diagnosis, prevention, monitoring, treatment, or mitigation of disease; the diagnosis, monitoring, treatment, mitigation, or compensation for injury or disability; the investigation, replacement, or modification of anatomical structures or physiological processes; the control of conception; and which do not achieve their primary intended effect in or on the human body through pharmacological, immunological, or metabolic means, but may assist their function through such means.

[0122] Medical devices vary depending on their intended use and indications. Examples range from simple devices such as tongue depressors, medical thermometers, and disposable gloves to advanced devices such as computers, implants, and prostheses that assist in medical testing.

[0123] According to the FDA's definition, a medical device is "an instrument, apparatus, tool, machine, device, implant, in vitro reagent or other similar or related article, including components or accessories: or recognized by the United States Pharmacopeia or any supplement thereof, intended for the diagnosis of a disease or other condition in a person or animal, or for the cure, relief, treatment or prevention of a disease in a person or animal, or intended to affect the structure or any function of the body of a person or other animal, and not to achieve any of its primary intended purpose through chemical action within or on the body of a person or other animal, and not in dependence on metabolism to achieve any of its primary intended purpose."

[0124] The current particles are not metabolized, nor do they have significant chemical effects in the body. These particles are radioactive carriers designed not to be metabolized or produce any chemical effects in the body, which makes the side effects (such as toxicity) of radiation therapy very limited.

[0125] Therefore, in one implementation, the term "medical device" should be understood to have the above FDA definition.

[0126] Imaging

[0127] Cancer imaging is a general term encompassing many methods used to study and diagnose cancer. Originally used for diagnosing and staging disease, cancer imaging is now also used to assist surgery and radiation therapy to look for early responses to cancer treatment and to identify patients who do not respond to treatment. One element is nuclear imaging, which involves the application of radioactive materials in disease diagnosis and monitoring. In a sense, nuclear medicine imaging is “radiology done from the inside out” or “internal radiology” because it records radiation emanating from within the body, rather than radiation produced by external sources such as X-rays. Furthermore, nuclear medicine scans differ from radiology because the focus is not on imaging anatomy but on function. Therefore, it is called a physiological imaging modality. Single-photon emission computed tomography (SPECT) and positron emission tomography (PET) scans are two of the most common imaging modalities in nuclear medicine.

[0128] In nuclear medicine, the approach that combines diagnostic imaging and treatment is called therapeutic diagnostics. Visualization of the distribution of radiolabeled particles helps predict whether a patient will benefit from treatment and / or determine the optimal dosage. While some therapeutic radionuclides possess decay properties that allow them to be imaged, only a few have decay properties that allow for specific and precise determination of the radioactive distribution within the body. Therefore, the idea arose to radiolabel reagents with alternatives to therapeutic radionuclides. For in vivo diagnostic imaging, short-lived gamma or positron emitters are typically used, such as… 99m Tc, 18 F and 67Ga. Despite possessing optimal imaging properties, the use of these radionuclides is not feasible in all cases due to their different chemical properties compared to therapeutic radionuclides. Furthermore, the significantly different half-lives between diagnostic and therapeutic radionuclides complicate the interpretation of biodistribution. Therefore, for example, 111 In has been used as 90 Alternatives to Y are available because they are both trivalent metals with similar half-lives (2.8 days and 2.7 days). The idea of ​​using a pair of radionuclides of the same element has also emerged, where one of the radionuclides has decay properties suitable for therapy, and the other is suitable for imaging—a so-called therapeutic diagnostic radionuclide pair. For example... 86 Y / 90 Y、 64 Cu / 67 Cu、 83 Sr / 89 Sr、 152 Tb / 161 Tb and 203 Pb / 212 Pb.

[0129] Therefore, one aspect of the present invention relates to the particles, compositions, or pharmaceutical compositions of the present invention for imaging. In one or more embodiments of the present invention, the imaging is nuclear medicine imaging.

[0130] combination therapy

[0131] Degradable particles can contain many different additional compounds. These compounds can be used for a variety of purposes, including targeting, stability, solubility, and degradation rate.

[0132] In one embodiment of the invention, the particle comprises one or more compounds selected from: monoclonal antibodies, polyclonal antibodies, radioimmunoconjugates, immunoconjugates, chelating antibody conjugates, vitamins including folic acid and folic acid derivatives, peptides, microantibodies, and affinity molecules.

[0133] In one embodiment, the particle comprises an antibody, antibody fragment, or protein, peptide, or vitamin derivative (targeting conjugate) that has affinity for receptors, including antigens on tumor cells.

[0134] In another embodiment, the particles contain radiolabeled antibodies, antibody fragments, or proteins, peptides, or vitamin derivatives (targeting conjugates) that have affinity for receptors, including antigens on tumor cells. The labeled particles thus generate a general particle radiation field on the surface, and the labeled antibodies or analogs deliver a specific alpha particle dose to the tumor cells via receptor or antigen binding.

[0135] The radionuclides in this invention can be conjugated to target molecules via bifunctional chelating agents.

[0136] These chelating agents can be cyclic, linear, or branched. Polyaminopolyacid chelating agents, which contain a linear, cyclic, or branched polyazine backbone with an acidic (e.g., carboxyl) group attached to the nitrogen atom of the backbone, may be mentioned in particular.

[0137] Examples of suitable chelating agents include DOTA derivatives, such as benzyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (p-SCN-Bz-DOTA) and tetra-primary amide variants of this DOTA compound, referred to as TCMC, and DTPA derivatives, such as p-benzyl-diethyltriaminepentaacetic acid (p-SCN-Bz-DTPA), the former being a cyclic chelating agent and the latter a linear chelating agent.

[0138] Metallization of the complexing portion can be performed before or after the complexing portion is joined with the target portion.

[0139] If the chelating agent is conjugated with the antibody before radiolabeling occurs, the radiolabeling procedure is generally more convenient in terms of timing and other options.

[0140] One aspect of the present invention relates to the particles, compositions or pharmaceutical compositions of the present invention used in accordance with the present invention, in combination with other cancer therapies.

[0141] Examples of treatments include chemotherapy, such as taxanes (e.g., paclitaxel, docetaxel), platinum-based drugs (e.g., carboplatin, cisplatin), doxorubicin, and mitomycin. Other examples are DNA repair inhibitors, such as PARP inhibitors (e.g., olaparib, rucaparib, niraparib, talaparib, veliparib, pamiparib, CEP 9722, E7016, and 3-aminobenzamide). A further example is radioimmunotherapy.

[0142] DNA repair inhibitors include ADP-ribose polymerase inhibitors (PARPi), MGMT inhibitors, DNA-dependent protein kinase inhibitors (DNA-PK inhibitors), ataxia-telangiectasia and Rad3-associated (ATR) kinase inhibitors, ataxia-telangiectasia mutation (ATM) kinase inhibitors, Wee1 kinase inhibitors, and checkpoint kinase 1 and 2 (CHK1 / 2) inhibitors.

[0143] In one or more embodiments of the present invention, PARPi is selected from olaparib, rucaparib, niraparib, talaparib, veliparib, pamiparib, CEP 9722, E7016 and 3-aminobenzamide.

[0144] PARPi can be olaparib. PARPi can be rucapranib. PARPi can be nirapanib. PARPi can be tarazobal. PARPi can be velipanib. PARPi can be pamiparib. PARPi can be CEP 9722. PARPi can be E7016. PARPi can be 3-aminobenzamide.

[0145] Particle preparation methods

[0146] One aspect of the invention relates to a method for preparing particles according to the invention, the method comprising contacting a degradable compound, a radionuclide, and a phosphorus-containing additive with or without a radionuclide carrier. The degradable compound and the radionuclide may form a first particle in an initial step, followed by an additional step of adding the degradable compound to the already radiolabeled particles to obtain layered particles. The phosphorus-containing additive is then added to stabilize the particles. The phosphorus-containing additive may become part of the particles in this process and remain in the composition containing the particles. Alternatively, the phosphorus-containing additive may be both part of the particles and remain in the composition containing the particles. Examples of different particle formulations can be seen in Examples 1 and 5. Another example of a particle formulation can be seen in Example 18.

[0147] Phosphorus-containing compounds such as phosphonates and phosphates can be used as additives to stabilize crystalline particles; these are the phosphorus-containing additives described herein. They can be added to particles, such as crystalline particles, during particle formation, after particle formation, after labeling, or to the final formulation to achieve size control. The product can be sterilized by autoclaving, and the additives can be added before or after this process. Size control additives can also be used with kit components used to prepare the final product.

[0148] Example 18 describes the production of smaller particles (CaCO3 SMP) by adding glycerol to a spontaneous precipitation reaction to generate calcium carbonate microparticles.

[0149] Therefore, in the embodiments, additional size control additives are added during particle preparation. In the embodiments, the additional size control additives are alcohols, such as, but not limited to, glycols or triols. In the embodiments, the additional size control additives are selected from ethylene glycol (EG), PEG, glycerol and dextran, and ethanol. In an example, the additional size control additive is glycerol.

[0150] Phosphorus-containing compounds can stabilize monodisperse and polydisperse crystalline particles. These particles are usually polydisperse because they are made in solution, but they can share characteristics, such as having sizes within a given similar range.

[0151] Therefore, one aspect of the present invention relates to the use of phosphorus-containing additives for controlling the size of particles containing degradable compounds. Another aspect of the present invention relates to the use of phosphorus-containing additives for stabilizing particles containing degradable compounds. These particles may be the particles of the present invention before or after the addition of a radionuclide. Therefore, the particles may be the degradable compound itself. In one embodiment of the present invention, the degradable compound is a grain.

[0152] Solutions or compositions containing radioactive nuclides, for example 224 Ra solution or composition with offspring 212 Mixtures of Pb can be treated with complexes prior to particle labeling. 212 Pb chelating antibody conjugate pretreatment to produce a two-component therapeutic system containing [amount of ingredients for use in the treatment of Pb]. 212 Radioimmunoconjugates and alpha emitters for Pb antigen-specific therapy, such as 224 Ra-labeled particles are used for general cavity therapy. Biological compounds, such as antibodies, can also be part of these particles. These compounds can then be mixed with phosphorus-containing additives to obtain particles with controllable size. The particles of the composition can also be... 212 In the composition specifically treated with Pb antigen 212 Pb-labeled particles.

[0153] One implementation involves a three-component system or kit containing a radionuclide, such as a radioimmunoconjugate for antigen-specific therapy, a degradable compound, and a phosphorus-containing additive.

[0154] The preferred method of using it is to use a kit containing a vial A containing a chelating conjugate antibody and a vial containing a radionuclide, such as one equilibrated with a daughter nuclide. 224 A vial B containing Ra and a vial C containing microparticles are used to add the contents of A to vial B, or vice versa. The mixture is incubated for a few minutes to a few hours, then transferred to vial C for further incubation for a few minutes to a few hours. The mixture is then mixed with a phosphorus-containing additive, transferred to a syringe, and injected into the patient.

[0155] This principle can significantly reduce the amount of treatment required, for example... 212 The content of Pb-radioimmunoconjugate, because it is expected 224 Ra-particles contribute significantly to the antitumor activity of such systems.

[0156] Another aspect of the invention relates to a kit comprising nanoparticles or microparticles according to the invention, and instructions for using the kit optionally.

[0157] In one embodiment of the invention, the kit comprises a chelating agent conjugate molecule, including a monoclonal antibody.

[0158] Because radionuclides have a half-life of several days, current methods and products allow for centralized production and delivery to end users. Another aspect of the invention is the use of biodegradable particles that slowly dissolve into calcium and carbonates, thereby producing small amounts of products already present in large quantities in the body. It is also noteworthy that the following feature is applicable when radionuclides, such as… 224 When Ra is absorbed by biodegradable compounds, such as calcium carbonate, it releases large amounts of short-lived compounds. 220 Rn(t 1 / 2 =56s), which will be associated with an ultra-short lifespan. 216 Po(t 1 / 2 =0.16s) decays into a longer-lived β emitter. 212 Pb(t 1 / 2 Two α particles are produced before (=10.6h). Lead, for example, has a very high precipitation ability in calcium carbonate, therefore in the ip fluid... 212 Pb will tend to recombine with particles, thereby reducing 212 Pb leaks into the systemic circulation.

[0159] Therefore, this is a very special technical feature, namely 224 Ra decays into a gas that can diffuse out of the particles and then further decay into... 212 Pb precipitates along with calcium carbonate.

[0160] Pre-formed particles and subsequent surface deposition or co-deposition of radionuclides to contain the radionuclides more deeply are two methods that can be used to produce therapeutic products. Once the radionuclides and degradable compounds form the first particles, phosphorus-containing additives can then be used to control the size of the first particles and form the particles of the present invention.

[0161] The particle sizes can range from nanometers to tens of micrometers, and radiolabeling is performed with high labeling yields. Furthermore, the particles can be stored for several days, which is important because it allows for the centralized production of ready-to-use particle suspensions and their delivery to hospitals. This can be done without compromising particle integrity, thus ensuring that the particles remain intact and of the size required for their intended use.

[0162] One aspect of the invention relates to particles produced by any of the methods described herein.

[0163] General Rules

[0164] It should be understood that any features and / or aspects discussed above with respect to compounds according to the invention are applicable by analogy to the methods described herein.

[0165] The following embodiments are provided to illustrate the present invention. These embodiments are intended to be illustrative and should not be construed as limiting in any way. Attached Figure Description

[0166] Figure 1 Intraperitoneal administration 224 Different variants of Ra-CaCO3 particles 1 day later 224 Ex vivo tissue distribution of Ra: 1 and 5 mg surface-labeled additive-free (A), 1 mg, 2 mg, 6 mg and 12 mg surface-labeled and EDTMP layer protection. 224 Ra-CaCO3- particles (B), surface-labeled as 224 Ra-CaCO3- particles were washed with different amounts of SHMP (C), and the inclusions in EDTMP were labeled as follows. 224 Ra-CaCO3-particles (D). In addition to the symbols representing the individual data points for each mouse, the data are displayed as bar graphs representing the median percentage injection dose per gram of tissue collected from organs and tissues.

[0167] Figure 2 Intraperitoneal administration of cationic drugs 224 Distribution of radioactive material in ex vivo tissues 1 day after Ra. Apart from the symbols representing individual data points for each mouse, the data are displayed as bar graphs representing the median percentage of injected dose per gram of tissue collected from organs and tissues.

[0168] Figure 3 Kaplan-Meier survival diagram of athymic Foxnu nude mice, with 1×10⁶ mice injected intraperitoneally. 6 One ES-2 cell was injected intraperitoneally with 0.9% NaCl or 1,000 ES-2 cells one day later. 224 Five different variants of Ra-labeled CaCO3 particles. N = 5-6 animals per group.

[0169] Figure 4 Kaplan-Meier survival plot of BALB / c mice, with mice intraperitoneally inoculated with 5×10⁶ mice. 4 One CT26.WT cell was administered via intraperitoneal injection one day later with 0.9% NaCl or 224 Four different variants of Ra-labeled CaCO3 particles. N = 7-8 animals per group.

[0170] Figure 5 Size distribution of CaCO3 particles. (a) Unlabeled, non-radioactive CaCO3-MP (MP: particles), suspended in brine and autoclaved on day 0, showed a different EDTMP% (w / w) compared to unautoclaved CaCO3-MP used as raw material. (b) Surface area of ​​unautoclaved CaCO3-MP compared to unautoclaved CaCO3-MP used as raw material.224 Ra-labeled CaCO3-MP has different EDTMP% (w / w). (c) Compared with unautoclaved CaCO3-MP used as raw material, autoclaved and layered CaCO3-MP has higher EDTMP% (w / w). 224 Ra surface-marked CaCO3-MP have different EDTMP% (w / w). (d) Comparison of surface-marked and layer-encapsulated surface-marked M, excerpted from (b) and (c) Dv: volume diameter, Dv10 is the 10th percentile, etc.

[0171] Figure 6 Sedimentation rates of non-radioactive autoclaved CaCO3 particles of different sizes. (a) Comparison of simulated surface-labeled MP suspended in 0.9% NaCl containing and without EDTMP after the suspensions were allowed to stand for 30 seconds and 4 minutes, respectively. (b) Turbidity assessment of suspended CaCO3-MP of different sizes, all simulated surfaces were labeled; layered MP is indicated by lines marked with "x". Dv: volume diameter, Dv50 is the 50th percentile.

[0172] Figure 7 CaCO3 particles (MP) adsorbed with different EDTMP concentrations 212 Pb and 224 Radiochemical analysis of Ra. Symbols represent independent samples, and EDTMP concentration is correlated with CaCO3. (a) Adsorption on MP at different days after labeling. 212 (a) Percentage of Pb (%RCP), MP surface-marked versus MP surface-marked in layered encapsulation. (b) Adsorption on MP at different days after marking. 224 Ra fraction (%RCP), MP of surface marking versus MP of surface marking in layered encapsulation. (c) Subtract 0.9% NaCl from the ITLC settings at different days after marking. 212 After the non-specific migration of Pb2+, in the liquid phase 212 The percentage of Pb-EDTMP, the MP marked on the surface and the MP marked on the layered package surface.

[0173] Figure 8 One day after IP injection, layer encapsulation. 224 Biodistribution of Ra-labeled CaCO3 particles. Bars represent medians, and symbols represent individual animals. Animals treated with equal or near-equal mass doses were grouped together. (a) 224 Ra is the percentage of the injection dose per gram of tissue. (b) From the same sample as in (a). 212 Pb percentage of the injected dose per gram of tissue. 224 A skull from the RaCl2 group 224 Ra and 212Pb data is missing; one blood sample from the 12mg group... 212 Pb data is missing.

[0174] Figure 9 In use 224 After Ra surface marking 208 Pb and 212 Distribution of Pb over days on layer-encapsulated microparticles containing 2.5% (w / w) EDTMP. Assume... 224 Ra and its daughter bodies are in equilibrium on day 0. 224 Ra、 212 Pb and 208 The total amount of Pb is 200 atoms (or au). Assume it is adsorbed on MP(RCP). 208 Pb and 212 The ratio of Pb to that in the solution was equal, with 94% RCP on days 0-3, 81% RCP on days 4-6, and 79% RCP on day 7. 224 Ra decay produces a 208 Pb atoms, and 212 The total number of Pb atoms was determined using the Bateman equation, taking into account... 224 Ra and 212 Pb decay.

[0175] Figure 10 . 212 Pb2+ and 212 The percentage of adsorption of Pb-EDTMP for non-radioactive simulated labeled CaCO3-MP, surface-labeled MP, and layer-encapsulated surface-labeled MP. EDTMP concentration represents the relative concentration of MP in the suspension relative to g / CaCO3.

[0176] Figure 11 High-pressure sterilization and 212 Radiochemical properties of various CaCO3 particles in the Pb-labeled suspension. Median diameter based on laser diffraction measurements. (a) Addition 212 The percentage of radiolabeled yield over time after Pb (same as the amount of RCP). (b, c) The percentage of RCP varies with particle size and / or phosphonate type on the same day as labeling (b) or at least 21 hours after labeling (c). (d, e) After incubation in an isotonic solution containing human serum albumin, 212 Pb is preserved in the particle type in (b,c).

[0177] Figure 12 . 212 Biodistribution of Pb was assessed as the percentage of the injected dose per gram of tissue at a specified time following intraperitoneal injection. Bars represent medians, and symbols represent individual animals. (a, b, c) Pamidronate in212 Pb-CaCO3 particles and 212 Free PbCl2 after administration 212 Pb 2+ Compared to (a) the percentage of injected dose per gram of tissue after 2 hours, (b) 6 hours, and (c) 24 hours. (d) pamidronate 212 The biodistribution of Pb-CaCO3 particles was compared with that of smaller particles (SMPs) in pamidronate produced in the presence of glycerol. No data on urine and / or bladder were reported in (a, c, d) due to missing data for two-thirds of the mice in one group, while one urine sample was missing in (b, d).

[0178] Figure 13 Survival figures of athymic Foxnu nude mice. The mice were inoculated intraperitoneally with ES-2 cells and injected intraperitoneally one day later with 0.9% NaCl or different active doses of pamidronate. 212 Treatment with Pb-CaCO3 MP. Detailed Implementation

[0180] Example 1 – Preparation of Particles

[0181] Subsequently used 224 Ra-labeled crystalline CaCO3 particles were prepared by spontaneous precipitation according to the scheme described by Volodkin et al. 2004. Equal volumes of 0.33 M Na2CO3 (Merck, Darmstadt, Germany) and 0.33 M CaCl2 (Merck) were used to precipitate the particles using a top-mounted stirrer (Eurostar 20) before centrifugation. (Werke GmbH & Co. KG, 120 Staufen, Germany) Vigorously mixed. The precipitate was washed in PH. Eur water and dried in a heated vacuum oven. The particles were predominantly spherical in shape, with a median diameter of 4 to 7 μm when measured using a laser diffraction particle size analyzer (Mastersizer 3000, Malvern Instruments Ltd, Worcestershire, UK).

[0182] As further described in Example 5, a stock solution of Na₂CO₃ and CaCl₂ up to 1 M was also prepared by mixing it with the existing... 224 Ra is mixed to prepare microparticles to produce inclusion body-labeled microparticles. Example 5 also describes the operation in some applications; a layer of CaCO3 is deposited on the surface of the labeled microparticles to form a thin encapsulation layer that protects the surface.

[0183] Example 2 - Controlling Size Using Phosphorus-Containing Additives

[0184] Phosphorus-containing compounds such as phosphonates and phosphates can be used as additives to stabilize crystal particles. They can be added to crystal particles during, after, or after labeling of crystals, or to the final formulation to achieve size control. Products can be sterilized by autoclaving, and additives can be added before or after this process. Size control additives can also be used as components of kits for preparing the final product.

[0185] Phosphonates and phosphonic acids are organophosphorus compounds containing C-PO(OH)2 or C-PO(OR)2 groups (where R = alkyl or aryl). Phosphonic acids, typically treated as salts, are generally non-volatile solids that are poorly soluble in organic solvents but soluble in water and common alcohols. Bisphosphonates include etidronate (HEDP), chlorophosphonate, tiludronate, pamidronate, neridronate, opaldronate, alendronate, ibandronate, risedronate, and zoledronic acid. Polyphosphonates include EDTMP-ethylenediaminetetra(methylenephosphonic acid), DOTMP-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacyl-tetra(methylphosphonic acid), and DTPMP-diethylenetriaminepenta(methylenephosphonic acid).

[0186] Phosphoric acid in the general sense is an oxyphosphate, in which each phosphorus atom is in the +5 oxidation state and is bonded to four oxygen atoms, one of which is arranged in a tetrahedral corner through a double bond. (The text then abruptly shifts to a seemingly unrelated topic: removing the H atom as a proton.) + The hydrogen atoms in the phosphate group convert phosphoric acid into phosphate anions. Partial removal produces various hydrogen phosphate anions. Phosphates include orthophosphates, linear oligophosphates, and polyphosphates, such as pyrophosphates, tripolyphosphates, and triphosphonic phosphates, as well as cyclic polyphosphates such as sodium hexametaphosphate (SHMP).

[0187] Example 3- 224 Ra and 212 Pb production

[0188] By 228 The Th source is mixed with actinide resin and loaded onto a column to prepare... 224 Ra generator. In 1MHNO3 228 The source was purchased from Eckert & Ziegler (Bruenk, Germany) or Oak Ridge National Laboratory (Tennessee, USA), based on The actinide resin used as the extraction solvent was purchased from Eichrom Technologies LLC (Lyre, Illinois) in 2 mL pre-packaged cartridges. Material was extracted from the actinide resin column, and the resin was pretreated with 1 M HCl (Sigma-Aldrich). Approximately 0.25 mL of actinide resin, 0.25 mL of 1 M HCl, and 0.1 mL of [unspecified solvent] were prepared in vials (4 mL vials, EC samples, Wheaton, Millville, NJ). 228 The slurry was prepared in 1M HNO3 and incubated with gentle stirring to fix it at room temperature. 228 Then, let it stand for 4 hours, and then let it stand for several days. The generator column was prepared in a 1 mL filter column (Isolate SPE, Biotage AB, Uppsala, Sweden), first using 0.2 mL of inactive actinide resin, and then containing... 228 The Th portion is loaded at the top. If released during generator operation... 228 Th, then an inert resin is introduced to the bottom of the column to act as a trapping layer. Afterwards, the generator capacity is increased. As described above, a mixture of 0.4 mL actinide resin and 0.5 mL... 228 A slurry consisting of 1 M HNO3 solution and 0.5 mL 1 M HCl was then loaded onto the generator column.

[0189] Radium-224 was periodically eluted from the generator column in 1-2 mL of 1M HCl. For further purification, the crude eluent from the generator column was loaded directly onto a second actinide resin column. The second column was washed with 1M HCl. This eluent was evaporated to dryness in a closed system. The vial was placed in a heater block and flushed with N2 gas through the Teflon tube inlet and outlet in the rubber / Teflon diaphragm of the vial. Acid vapor was introduced into a saturated NaOH beaker via a flow of N2 gas. The remaining radioactive residue after evaporation was dissolved in 0.2 mL or larger of 0.1M HCl. A radioisotope calibrator (CRC-25R, Capintec Inc., Ramsey, NJ, USA) was used to measure the total activity extracted during the process.

[0190] Lead-212 is made from 224 Ra passed 220 Rn radioactivity was generated using a novel, simplified single-chamber diffusion system, an improvement upon the method described by Hassfjell S., 2001. Approximately 2-20 μL of the solution prepared as described above was used. 224RaCl2 solution (~250 kBq) was distributed on the surface of a small paper strip (15 × 5 mm) attached to the tip of a syringe, which had previously been inserted through the silicone diaphragm of a 3 mL microreactor glass V-shaped vial (Supelco, Darmstadt, Germany) with a screw cap. The screw cap was carefully attached to the V-shaped vial, avoiding contact between the paper strip and the inner surface of the vial. The sealed V-shaped vial was left to stand overnight in a fume hood. Next, 220 Rn from 224 The Ra source is released through the air inside the vial, via its short-lived daughter particles. 216 Po diffuses and decays into 212 Pb was deposited in the container. After 20 to 28 hours, carefully remove the screw cap with the paper strip attached, avoiding... 224 Ra contaminated vials. Then, 1M HCl was used to remove the contaminant. 212 Pb was washed off the glass wall and transferred to a new container.

[0191] Example 4 - Method for measuring radioactivity

[0192] Radioactive samples up to approximately 30 kBq were measured using an automated NaI gamma counter (Hidex, Turku, Finland) in the energy range of 60–110 keV. 212 Pb, and measurements in the energy range of 65-345 keV. 224 Ra. As shown in Table 1, the most abundant x and γ radiation in these energy ranges comes from 212 Pb. Assume the counts in these windows primarily come from... 212 Pb, with other nuclides in this series contributing very little. Radium-224 activity was determined indirectly based on counts within a 65–345 keV window. This was performed by re-measuring the sample 2–4 days after the initial measurement, at which point the initial activity present in the sample was considered. 212 Pb has decayed and a transient equilibrium has been established. According to 224 Ra and 212 The half-life of Pb, pure 224 The Ra source reached equilibrium after about 2 days.

[0193] Table 1 224 Overview of X-rays and / or gamma rays with an abundance of 1% or higher in the Ra series. X-rays and gamma rays are divided into two columns: one for energies between 65 and 345 keV, and the other for energies above 345 keV. Radionuclides 60-110 keV > 110 keV

[0194] (Abundance, %) (Abundance, %)

[0195]

[0196]

[0197] The radioisotope calibrator (CRC-25R or CRC-55tR Capintec Inc., Ramsey, USA) is used to measure samples above 50 kBq.

[0198] To determine the sample 224 Ra and 212 Real-time Pb distribution was determined using a high-purity germanium detector (HPGe) (BroadEnergy germanium detector BE383OP or standard electrode coaxial germanium detector GC3518, Mirion-Canberra, USA). The spectral analysis software package used with the instrument was based on the Genie algorithm.

[0199] Example 5 - Using 224 Ra labeling: Ra-224 labeled CaCO3 particles were prepared through three different procedures: surface labeling, inclusion body labeling, and layer protection.

[0200] 1. By 224 Ra is adsorbed onto the surface of pre-prepared CaCO3 particles for surface labeling.

[0201] 2. By using CaCO3 particles during their formation process... 224 Ra is incorporated into CaCO3 particles for inclusion body labeling.

[0202] 3. Surface-labeled CaCO3 particles: After radioactive labeling, a layer of CaCO3 precipitates onto the original surface to encapsulate the radionuclide.

[0203] Regarding surface labeling, CaCO3 particles were washed three times with water and twice with 0.1M Na2SO4 (Merck) immediately before radiolabeling. The CaCO3 particles had a Ba content of 0.004 w / w% relative to CaCO3. 2+ and 0.6 w / w% SO4 2- In the presence of Merck, a radium-224 solution (in 0.1 M HCl and 0.035–0.5 M NH4OAc) was added to the particles for use in... 224 Ra2+ co-precipitated on the surface of the particles. The radiolabeling process was carried out in a 0.9% saline (Merck) solution with orbital rotation for 1.5 hours (HulaMixer, Invitrogen, Thermo Fisher Scientific, Massachusetts, USA).

[0204] Inclusion body-labeled CaCO3 particles were prepared by: using a mixture containing 0.004 to 0.3 w / w% Ba... 2+(relative to CaCO3) and the required amount 224 Quickly pour the required volume of 0.33M or 1M CaCl2 solution into an equal volume of 0.33M or 1M CaCl2 solution containing 0.6 to 0.7 w / w% SO42-. 2- In a Na2CO3 solution (relative to CaCO3), mix using a magnetic stirrer (BioSan MS3000, Riga, Latvia) or by hand shaking and vortexing. In some applications, 1 mg / ml gelatin (Sigma-Aldrich) is added to the CaCO3 crystalline phase to slow crystallization and the mixture is stored overnight in a refrigerator.

[0205] use 224 Ra was used to surface-label the CaCO3 particles protected by the coating, and excess labeling solution was removed. Subsequently, the particles were dispersed in CO3 containing a concentration of 0.33–0.66 M under vigorous stirring. 2- and Ca 2+ In the solution of ions, a thin layer of CaCO3 is precipitated to cover the original surface.

[0206] In all cases, excess layering or labeling solution was removed before the CaCO3 particles were washed once or twice with 0.9% NaCl, and then dispersed in NaCl or NaCl supplemented with a phosphorus compound. In some cases, when two washing steps were performed, a phosphorus compound was added after the first washing step to provide an additional wash for the particles. Finally, the radiolabeled particle suspension was autoclaved.

[0207] Example 6 - Controlling the size of CaCO3 particles using EDTMP, pamidronate, and SHMP

[0208] Calcium carbonate microparticles were washed three times with water for injection (WFI), and in some cases twice additionally with 0.1 M sodium sulfate, and then suspended in solutions of different concentrations of additives (EDTMP, pamidronate, or SHMP) containing 0.9% NaCl. For comparison, samples with no additives were suspended only in 0.9% NaCl. Particle concentrations ranged from 30 to 250 mg / ml. The suspensions were autoclaved at 121°C for 21 minutes. In some cases, as described in Example 5, samples with surface markings were... 224 The CaCO3 particles of Ra were measured.

[0209] The particle size of the autoclaved suspension was measured using a laser diffraction particle size analyzer (Mastersizer 3000, Malvern Instruments Ltd, Worcestershire, UK) until the suspension was prepared and stored at room temperature for 8 days.

[0210] Table 2 summarizes the additives and their concentrations, particle size distribution, and measurement dates. For comparison, the median volume diameter of the untreated CaCO3 particle feedstock suspended in water is also given. Without additives, sample preparation immediately resulted in a 3-fold increase in median particle size, and the particles were even larger after 5 days due to continued growth in the aqueous medium. With the addition of additives EDTMP, SHMP, or pamidronate, the particle size was stabilized relative to the untreated feedstock. The additive concentration had the most significant effect on EDTMP; while a concentration of 0.001 g / g relative to CaCO3 did not stabilize the particle size 5 days after sample preparation, at EDTMP concentrations >0.013 g / g, the size remained stable for at least 8 days.

[0211] Table 2. Effects of different phosphorus additives on the particle size distribution of CaCO3 after autoclaving.

[0212]

[0213]

[0214] (a) Surface-marked particles, *free of 224 The simulated labeling process of Ra.

[0215] (b) Includes an additional washing step with WFI after the addition of the additive to remove any unbound additive, which may affect the final additive concentration.

[0216] (c) Measured on day 13.

[0217] Example 7 - In vitro retention of radioactivity on CaCO3 particles: The effect of phosphorus compounds as additives

[0218] Radiolabeling of CaCO3 particles was performed as described in Example 5, including surface labeling, inclusion body labeling, or layer-protected radiolabeling. The particles were suspended at a concentration of 12.5-250 mg / ml in 0.9% NaCl with relevant additives (EDTMP, pamidronate, or SHMP) relative to CaCO3, with a concentration range of 0-0.100 g / g. The suspension was autoclaved and left at room temperature. To determine... 224 Ra and 212 The retention of Pb on the microparticles was assessed by taking samples at different time points after autoclaving and dividing them into equal portions. The microparticles were then separated from the liquid phase by centrifugation, and the radioactivity in the precipitate and supernatant fractions was measured using an HPGe detector or a Hidex automated gamma counter (see Example 4 for details). The percentage of radioactivity retained on the microparticles was defined as the ratio of the activity in the precipitate fraction before separation to the activity in the entire sample.

[0219] Tables 3 and 4 show the surface-marked and layer-protected CaCO3 particles, respectively. 212 Pb and 224 Overview of Ra's retained radioactivity. In the first week after sample preparation, in addition to... 212 Aside from the trend of lower Pb retention and higher EDTMP levels, the addition of phosphorus compounds did not affect the percentage of radioactivity retained on the particles. The addition amounts tested were able to control the particle size (see Example 6).

[0220] Table 3. Effects of different phosphorus additives on the retention of radioactivity on CaCO3 particles labeled on the in vitro surface.

[0221]

[0222]

[0223] (a) Includes additional centrifugation after the addition of the additive to remove any excess that may affect the final additive concentration.

[0224] Table 4 shows the effect of EDTMP on radioactive in vitro retention on CaCO3 particles protected by a layer.

[0225]

[0226] Example 8 - Intraperitoneal administration 224 Biodistribution of Ra-labeled CaCO3 particles in mice

[0227] Healthy female athymic Foxn raised in a facility nu Nude mice. Intraperitoneal administration (IP) to mice. 224 Ra-CaCO3- particles or cations 224 Different variants of Ra (see Table 5 for details) were administered, and blood was collected via cardiac puncture approximately one day after injection while the animal was anesthetized. The animal was then immediately euthanized before the selected organs and tissues were collected, weighed, and the radioactivity measured using a Hidex gamma counter. The estimated radioactivity per gram of tissue was... 224 Ra and has the longest half-life 212 The percentage of Pb (10.6 h) daughter nuclide injected. Samples were measured as soon as possible after sacrifice for estimation. 212 The amount of Pb, and a remeasurement at least 2 days after sacrifice, is used to determine 224 The amount of Ra. The measured radioactivity is directly compared with the radioactivity in a standard injectable sample measured together with the same sample.

[0228] Table 5 uses cations 224 Ra and 224An overview of biodistribution experiments on different variants of Ra-labeled CaCO3 particles.

[0229]

[0230]

[0231] MP: microparticles

[0232] In vitro biological distribution data indicate that tissue distribution is used in ( Figure 1 (B, C, and D) and not using ( Figure 1 A) Size control additives 224 Ra-CaCO3 particles are fundamentally similar. (The text abruptly shifts to a seemingly unrelated topic: "and the cations administered via IP...") 224 Compared to Ra, the use of phosphorus-containing additives 224 All variants of Ra-CaCO3 particles showed increased IP radioactivity retention, which was reflected in reduced absorption in the femur and skull. Figure 2 For variants without additives () Figure 1 A) After injecting 1mg, bone density... 224 Ra absorption and IP injection of cations 224 The absorption after Ra is quite ( Figure 2 ),and 224 Ra significantly improved IP retention in the 1 mg protective layer. Figure 1 B).

[0233] Example 9 - Therapeutic effect of radiolabeled microparticles dispersed in phosphorus compounds in a mouse ovarian cancer xenograft model

[0234] Human ovarian cancer often causes ascites, so the therapeutic effect of radiolabeled microparticles dispersed in phosphorus compounds was examined in an ovarian cancer xenograft model that induces invasive tumor cell growth and ascites formation in immunodeficient mice.

[0235] Xenografts are produced by feeding 4-5 week old female athymic Foxn cells in an institution. nu A single intraperitoneal injection of ES-2 cell suspension (1×10⁶ cells in 0.2 ml RPMI) into nude mice 6 Produced by (a few cells). Approximately one day later, it is administered via intraperitoneal injection. 224Mice were treated with different variants of Ra-labeled CaCO3 microparticles (Table 5). Mice in the control group were administered 0.9% NaCl via intraperitoneal injection (IP). Efficacy was assessed by survival time. Mice were monitored for changes in body weight, behavior, posture, and appearance at least twice weekly, with more frequent monitoring when they showed signs of disease progression. Mice were euthanized by cervical dislocation when they reached predetermined endpoints, including rapid weight loss (>10% within one week), severe mobility impairment due to ascites accumulation, and / or cachexia.

[0236] Survival time is recorded as the number of days after tumor cell inoculation. The Kaplan-Meier survival curve is shown below. Figure 3 As shown, the median survival time is as follows Figure 6 As shown. Comparison of survival curves showed that all experimental groups were significantly different from the control group (paired log-rank test, p≤0.0014, familial significance level 0.05). On day 19 after tumor cell inoculation, all mice in the saline control group were euthanized, while... 224 None of the mice in the Ra-labeled CaCO3 microparticle treatment group reached the study endpoint. Compared to the saline control, all... 224 Ra treatments all showed similar survival extensions, regardless of whether phosphorus additives were used or which phosphorus additives were used (EDTMP vs. SHMP) or whether the particles were surface-labeled or inclusion-labeled.

[0237] Table 6 summarizes the selected study details and results: 224 The efficacy of Ra-CaCO3- microparticles in the ES-2 human ovarian xenograft model in immunodeficient nude mice.

[0238]

[0239] 1 Radioactivity administered to each mouse was based on measurements taken five days post-injection using a Hidex automated gamma counter on three standard samples from each group. Measurements for each standard were attenuated to the injection time, and these means and corresponding standard deviations are reported. *Statistically significant compared to the saline control group, with a family significance level of 0.05. Survival curves were compared pairwise using a log-rank test and adjusted for multiple comparisons using the Bonferroni method.

[0240] Example 10 - Antitumor efficacy of radiolabeled microparticles dispersed in phosphorus compounds in a mouse model of homologous colon cancer.

[0241] Colorectal cancer frequently leads to peritoneal metastasis; therefore, the therapeutic effects of radiolabeled microparticles dispersed in phosphorus compounds were examined in an immunocompetent mouse model of IP colon cancer.

[0242] A single intraperitoneal injection (IP) of the mouse colorectal cancer cell line CT26.WT (5 × 10⁻⁶) into approximately 6-week-old female BALB / cAnNRj mice (Janvier Labs, France) was performed. 4 Tumors were established using 100 cells in 0.2 ml PBS. One day later, an intraperitoneal injection was administered. 224 Mice were treated with different variants of Ra-labeled CaCO3 particles (Table 8). Mice in the control group were administered 0.9% NaCl via intraperitoneal injection (IP). Efficacy, as a measure of survival, was based on rapid changes in body weight, development of ascites, body appearance scores, and the development of palpable abdominal tumors. Mice were euthanized upon reaching these predetermined endpoints due to cervical dislocation.

[0243] Survival time is recorded as the number of days after tumor cell inoculation, and the Kaplan-Meier survival curve shows... Figure 4 The median survival time is shown in Table 7. The median survival time increased from 18 days in the control group to different... 224 Ra-CaCO3- particles for 27 to 33 days. Compared with the saline control, all 224 Ra treatments all exhibited similar survival extensions, regardless of whether a phosphorus compound was used or which phosphorus compound was used (EDTMP vs. SHMP) or whether the particles were surface-labeled or inclusion-labeled.

[0244] Table 7 summarizes the details and results of the selected studies: intraperitoneal administration in a BALB / c mouse homologous CT26.WT colon cancer tumor model. 224 The efficacy of Ra-labeled CaCO3 particles.

[0245]

[0246] 1 Radioactivity levels in each mouse were determined based on measurements taken three days post-injection using a Wizard 2 gamma counter on three standard samples from each group. Measurements for each standard were decay-corrected to the injection time, and these averages and corresponding standard deviations were reported.

[0247] Example 11 - Lead-212 labeled calcium carbonate microparticles in pamidronate

[0248] Calcium carbonate particles and 212The Pb solution was prepared as described in Examples 1 and 3 above. Dry CaCO3 particles were suspended in WFI, sonicated, and washed a total of 3 times with WFI. Finally, the particles were suspended in 0.9% NaCl at a concentration of 25-50 mg / ml, and disodium pamidronate was added to the suspension to achieve a pamidronate / CaCO3 relative concentration of 0.01 g / g. The suspension was autoclaved at 121°C for 20 minutes in sealed sample vials and then cooled to room temperature. The pH of the lead-212 solution was neutralized by adding 5M ammonium acetate and 1M sodium hydroxide (10 v / v each). During radiolabeling, approximately 50% of the volume of the particle suspension was injected using a syringe. 212 Pb solution was added to a sealed container containing the particulate suspension, and the suspension was then vortexed and placed on a horizontal shaker for 3 to 60 hours. The activity per particulate mass varied between 3–99 kBq / mg. In some cases, the particulate suspension was further diluted with 0.9% NaCl after the labeling procedure.

[0249] The yield of radiolabeled particles was assessed by separating the liquid phase from small suspension samples (supernatant S and particulate precipitate P). Radioactivity in both portions was measured using an HPGe detector or a Hidex automated gamma counter. The yield of radiolabeled particles was assessed as CPM(P) / CPM(S) + CPM(P), where CPM represents the counts per minute.

[0250] Table 8 provides an overview of the radiolabeling yield. Horizontal shaking incubation time does not affect the radiolabeling yield, but the higher the activity concentration, the more pronounced the yield increase.

[0251] Table 8. CaCO3 particles with added pamidronate for size control 212 Yield of Pb labeling.

[0252]

[0253] (1) The average of two parallel radiolabeled samples.

[0254] Example 12 - Size control of radiolabeled calcium carbonate microparticles with and without calcium carbonate layer by adding EDTMP.

[0255] The size of unlabeled, analog-labeled, and radiolabeled CaCO3-MP (MP: microparticles) in suspensions with different concentrations of EDTMP was measured using laser diffraction (Mastersizer 3000, Malvern Instruments Ltd., Worcestershire, UK). Unautoclaved CaCO3-MP, used as the raw material for radiolabeling and analog labeling, was used as a reference by dispersing a small amount of dried CaCO3-MP in water and then sonicating it. The dimensional stability of radiolabeled CaCO3-MP over time was evaluated by measurements taken after 7 days of storage at room temperature; surface-labeled MP was compared with layer-encapsulated MP.

[0256] The ability of EDTMP to control the size of calcium carbonate particles was tested. The calcium carbonate particles were radiolabeled with radium-224 and encapsulated by adding a layer of calcium carbonate before the suspension was sterilized by autoclaving. Figure 5 Adding at least 1% EDTMP can control particle size.

[0257] This example demonstrates that the dimensional control achieved through EDTMP is also applicable to layered encapsulated products with radioactive markings.

[0258] Example 13 – Controlling the size of calcium carbonate particles by adding EDTMP to slow down the settling rate of particles in suspension.

[0259] The ability of MP to remain suspended in solution was assessed by sedimentation rate, which was studied through visual inspection of the samples and evaluation of the turbidity of different suspensions of non-radioactively labeled CaCO3-MP (with and without EDTMP). Turbidity was assessed by diluting the CaCO3-MP suspension with water (water for injection) and then measuring the change in optical density at 800 nm wavelength over 30 minutes using a spectrophotometer (Hitachi U-1900, Hitachi High-Tech, Tokyo, Japan). The 800 nm wavelength was chosen to reduce the potential light absorption of CaCO3 and improve light scattering by the particles. Therefore, the decrease in optical density over time was directly correlated with the decrease in light scattering of MP, and thus directly correlated with the decrease in turbidity of the sample due to sedimentation. See Figure 6 .

[0260] This example demonstrates that the sedimentation rate decreases as the particle size decreases. There are several improvements to the treatment of suspensions for clinical administration of the product.

[0261] Example 14 - Radiochemical properties dependent on EDTMP and layer encapsulation

[0262] Radiochemical purity is defined as the percentage of radionuclides retained on the phosphorus (MP) fraction after a certain period of time. A small suspension is separated into the MP fraction (P) and the supernatant fraction (S) by centrifugation. Percentage radiochemical purity, %RCP, is defined as the proportion of radioactivity in the P fraction:

[0263] CPM(P) / CPM(P+S), where CPM represents the count per minute. Radioactivity in both sections was measured using a Hidex automatic gamma counter (Hidex Oy, Turku, Finland). 212 The radioactivity of Pb is quantified by counting within a 60-110 keV window. For 224 Ra is indirectly determined by the following method: assuming that at least two days have elapsed between the two partial decays. 224 Ra and offspring 212 A transient equilibrium is reached between Pb and Pb, and then measurements are taken in the 65-345 keV window. 212 Pb activity, from which gamma energy and X-rays primarily originate, was assessed by repeated sampling and measurements after storage at room temperature for up to 7 days. 212 Pb and 224 Ra%RCP stability over time.

[0264] exist 224 Different variants of Ra-CaCO3-MP were evaluated in the liquid phase for the release of MP. 212 Complexation between Pb and EDTMP in solution ( Figure 7 First, the liquid fraction was separated from MP by centrifugation. Then, the concentration of MP in the obtained supernatant was measured using instantaneous thin-layer chromatography (ITLC) strips (Tec-Control Chromatography Systems #150-772, Biodex Medical Systems, Inc., New York, USA). 212 The degree of Pb-EDTMP complexation. Chelated 212 Pb will migrate with the mobile phase in the system, while most (>90%) will remain unbound. 212 Pb2+ will remain on the original line, thus allowing for evaluation. 212 Pb-EDTMP complexation. Water (pharmaceutical grade) or 0.9% NaCl was used as the mobile phase, and the strip was cut in half after reaching the top line at the solvent front. As previously described, the concentrations in both halves were measured using a gamma counter. 212 The radioactivity of Pb. The degree of chelation is determined by migration. 212 Pb in 224 The proportion of Ra-CaCO3-MP in the liquid portion is defined by subtracting the free EDTMP in a 0.9% NaCl solution. 212 Pb 2+The non-specific migration is quantified. Equation 1 describes the chelation percentage; A 212 Pb-EDTM indicates 224 The measured activity of Ra-CaCO3-MP in the supernatant was A. 212 Pb represents free phosphorus in a 0.9% NaCl solution. 212 Pb 2+ The measured activity is given, where m and o represent the two parts of the ITLC band, respectively; m represents migration with the mobile phase, and o represents the original band.

[0265]

[0266] This embodiment demonstrates 212 Pb and 224 A comparison of Ra retention on CaCO3 particles with and without layer encapsulation shows how adding a layer to the product in EDTMP can improve, particularly... 212 Pb is retained.

[0267] Example 15 - Encapsulation of EDTMP in IP Injection 224 Biodistribution of radium-224 and lead-212 after Ra-CaCO3-MP

[0268] The effect of EDTMP-added layer encapsulation was evaluated in institutionally housed female athymic nude mice (Hsd: Athymic Nude-Foxn1nu). 224 Biodistribution of Ra-CaCO3-MP. As previously described, calcium carbonate microparticles were labeled and autoclaved. The effect of mass dose (mg dose) was considered by testing dose ranges of 1–12 mg CaCO3 and 6–18 kBq using isotonic infusion solution (Plasmalyte, Baxter International Inc., Illinois, USA). One day after a single intraperitoneal administration, mice were euthanized by cervical dislocation and tissue samples were obtained for radioactivity measurement. Three standard samples corresponding to 25–50% of the administered dose for each treatment were used to determine the injected radioactive dose. The radioactivity of tissues and standard samples was measured using a gamma counter as described above. 212 Pb and 224 The radioactivity of Ra was used to calculate the percentage of injected dose per gram of tissue (%ID / g). For two reasons, the radioactivity of Ra was not considered when calculating %ID / g. 224 Ra and 212 Correction was made for Pb decay and / or inward growth. First, the counting time interval between standard samples and tissue samples was less than 2-3 hours (i.e., ...). 224 (3% of the Ra half-life), and secondly, when the measured activity is close to or below the device's quantitation limit, it can avoid [the risk of adverse reactions due to]... 224Error propagation caused by the uncertainty of Ra measurement values. As free tissue one day after IP injection. 224 Reference for Ra bone accumulation, a group of mice received approximately 30 kBq 224 RaCl2, as follows The preparation was carried out as previously described. See [previous description]. Figure 8 .

[0269] This example demonstrates a comparison of the biodistribution of radium-224 and lead-212 between different numbers of layered encapsulated microparticles and free radium-224. Bone resorption levels decreased with increasing microparticle count.

[0270] Example 16- 208 Pb and 212 Distribution of Pb on layer-encapsulated microparticles with 2.5% (w / w) EDTMP.

[0271] Chemothermic stable daughter nuclides adsorbed on MP 208 The accumulation of Pb increases over time, although this seems to have an effect on... 212 The adsorption of Pb has little effect. For the surface-labeled variant at this EDTMP concentration, 212 The %RCP of Pb was only 56% on day 0, increasing to 70% on day 4.

[0272] Example 17- 212 Pb 2+ and 212 Pb-EDTMP adsorbs onto calcium carbonate particles.

[0273] EDTMP and 212 The known complexing properties of Pb and calcium indicate that 212 The Pb-EDTMP complex may also associate with MP. To verify this hypothesis, in the addition of 212 After Pb-EDTMP solution, the following was evaluated. 212 The adsorption of Pb on non-radioactive simulated labeled CaCO3-MP with and without encapsulation was observed. The results showed that 17-20% of the adsorption was... 212 Pb-EDTMP adsorbs onto MP; when unbound MP is added... 212 Pb 2+ ( 212 When PbCl2 is used, the adsorption capacity increases to 96%. 212 The reduced adsorption of Pb-EDTMP is consistent with general observations, namely, at higher EDTMP concentrations in MP suspensions, the adsorption of surface-labeled MP and layer-encapsulated MP decreases. 212 The %RCP of Pb decreased.

[0274] Example 18 - Preparation of Smaller Particles (CaCO3 SMP)

[0275] Calcium carbonate microparticles were produced using two different procedures to generate particles with two different size groups. One procedure for producing CaCO3 microparticles is detailed in Example 1, while the other procedure is similar to the first, however, glycerol is added during the spontaneous precipitation reaction in an attempt to produce smaller microparticles (CaCO3SMP). Before mixing 1M CaCl2 and Na2CO3 (Merck) solutions, glycerol (Sigma-Aldrich) was added to each solution to a concentration of 50% (v / v), diluting the solutions to 0.5M. The two solutions were then combined by vigorous stirring for 30 minutes using an overhead stirrer running at 6000 RPM. The resulting CaCO3 SMP precipitate was washed three times with water for injection (WFI) before drying at 180°C. The particle size distribution was measured using a laser diffraction particle size analyzer (Mastersizer 3000, Malvern Instruments Ltd, Worcestershire, UK). The CaCO3 MP and CaCO3 SMP used in Examples 19, 20, and 21 have volume-based median diameters of approximately 5 μm and 2 μm, respectively. The process was carried out as described in Example 11. 212 Pb marker.

[0276] Example 19 - Autoclaved and used 212 Radiochemical properties of various CaCO3 particles in Pb-labeled suspension

[0277] 212 Measurements of Pb radiolabeled particles (generated as described in Example 1 or Example 18 and labeled as described in Example 11) included radioactivity concentration, radiochemical yield / purity, stability over time, and parent nuclide. 224 Determination of potential breakthroughs in Ra. Measurement of suspension using an ionization chamber dosimeter (Capintec Inc., New Jersey, USA). 212 The radioactivity levels of Pb solution and vials of labeled CaCO3 particles were measured. A small portion of the suspension was separated into a particle fraction (P) and a supernatant fraction (S) to measure the adsorption on the particles. 212 The Pb fraction was used to determine the percentage radiolabeled yield, %RCY. Both fractions were measured separately on an automatic gamma counter (Hidex Automatic Gamma Counter, Hidex Oy, Turku, Finland). %RCP was calculated as the ratio of CPM(P) / CPM(P+S), where CPM represents the count per minute in each fraction to indicate the amount adsorbed on the particles. 212 The Pb portion.

[0278] In vitro stability is defined as the retention on microparticles in an in vitro setting. 212The percentage of Pb, in this in vitro setup, included incubation at 37°C in isotonic infusion solution (Plasmalyte, Baxter, Illinois, USA). 212 Pb-CaCO3 MP / SMP incubation was performed for 1.5 to 21 hours, with the solution pH approximately 7 and supplemented with 10 g / L human serum albumin (Sigma-Aldrich, Missouri, USA). The CaCO3 concentration was then reduced from the initial 25 mg / ml to 2 mg / ml or 6 mg / ml. The diluted particles were separated from the incubation solution, and the radioactivity of both fractions was measured using the method described above for RCP determination.

[0279] In these experiments, all variants 212 Pb%RCP>90%, diluted at 2-6 mg / ml and incubated resulted in 212 The release of Pb into the incubation solution appears to be related to particle type and concentration. For the largest recrystallized particles and the lowest concentration, 212 Pb has the lowest retention rate on particles, and its contribution to the extended duration from 90 minutes to 21 hours is negligible. 212 Pb has a high retention rate on aragonite particles, and SMP may have higher stability compared to MP.

[0280] The results are as follows Figure 11 As shown.

[0281] Example 20 - Intraperitoneal administration 212 Biodistribution of Pb-labeled CaCO3 particles in mice

[0282] 212 Pb-CaCO3 MP and 212 Pb-CaCO3 SMP is labeled as in Example 11. It is administered intraperitoneally with 0.9% NaCl ( 212 Free PbCl2) 212 Pb 2+ In contrast, intraperitoneal administration in tumor-free mice 212 Following Pb-CaCO3 MP, the biodistribution of lead-212 was assessed. The biodistribution of 5 mg of lead-212 with a median particle size of 5 μm was investigated based on volume. 212 Pb-CaCO3 MP dose. Mice were sacrificed by cervical dislocation at 2, 6, and 24 hours post-treatment, and tissue samples were collected for radiometric measurements and calculation of the percentage of injected dose per gram of tissue (%ID / g). In a second experiment, 5 mg of Pb-CaCO3 was compared based on volume with a median diameter less than 3 μm. 212 Pb-CaCO3 MP and 212Biodistribution of Pb-CaCO3 SMP. Radioactivity measurements were performed using a Hidex automated gamma counter with appropriate calibration factors to obtain Bq data. 212 The 10.6-hour half-life of Pb was used to correct for decay in the measurements. Kidney and liver samples were remeasured after >24 hours of decay to ensure accuracy. 212 Instantaneous equilibrium between Pb and daughter products. During injection 212 Two hours later, high levels of PbCl2 were detected in the kidneys. 212 Pb was detected in large quantities in blood, liver, and bones. 212 Pb. For those bound to MP 212 Pb, %ID / g in these tissues and the spleen, was significantly reduced. Six hours later, in the kidneys of both variants... 212 Pb levels decreased, indicating clearance, but remained stable in bone and associated with these tissues. 212 Compared to PbCl2, it binds MP. 212 The %ID / g of Pb was significantly higher. After 24 hours, that is, exceeding... 212 Pb has two physical half-lives, a decrease in %ID / g in most soft tissues, and, except for the detection of no statistically significant difference between MP-bound and free variants in the skull (p<0.020), accumulation in bone... 212 Pb increases relative to earlier time points of MP.

[0283] The results are as follows Figure 12 As shown.

[0284] Example 21 - Intraperitoneal administration 212 Therapeutic effects of Pb-labeled CaCO3 microparticles on mice

[0285] In a mouse model of ovarian cancer, iP xenograft was performed. 212 Efficacy study of Pb-CaCO3 MP 1% pamidronate in the treatment of cavitary carcinoma. Nude mice were intraperitoneally injected with 300,000 ES-2 cells (ATCC, Wesel, Germany) and then administered a single intraperitoneal dose of 2-5 mg and 63-430 kBq the following day. 212 Mice were treated with Pb-CaCO3 MP. All radioactive doses were retrospectively measured and calculated from standard samples as described above. Control animals were given saline or unlabeled CaCO3 MP suspended in saline and 1% (w / w) pamidronate. Compared to both the saline control containing pamidronate and the unlabeled CaCO3 MP, all tested doses in pamidronate were observed to be significantly reduced. 212 Pb-CaCO3 MP showed significant therapeutic effects. For labeled MP, the effect appeared to be dose-dependent.

[0286] The results are as follows Figure 13 As shown.

[0287] item

[0288] 1. A particle comprising a degradable compound, a radioactive nuclide, and a phosphorus-containing additive.

[0289] 2. The particles according to claim 1, wherein the degradable compound is selected from CaCO3, MgCO3, SrCO3, BaCO3, calcium phosphate including hydroxyapatite Ca5(PO4)3(OH) and fluorapatite, and complexes having any of these as the main component.

[0290] 3. The particles according to items 1-2, wherein the degradable compound is CaCO3.

[0291] 4. The particles according to items 1-3, wherein the phosphorus-containing additive is a phosphate selected from orthophosphates, linear oligophosphates, polyphosphates, and cyclic polyphosphates.

[0292] 5. The particles according to items 1-3, wherein the phosphorus-containing additive is a polyphosphate selected from pyrophosphate, tripolyphosphate and triphosphonic phosphate.

[0293] 6. The particles according to items 1-3, wherein the phosphorus-containing additive is a cyclic polyphosphate, which is sodium hexametaphosphate (SHMP).

[0294] 7. The particles according to items 1-3, wherein the phosphorus-containing additive is a phosphonate.

[0295] 8. The particle according to item 7, wherein the phosphonate is a diphosphonate.

[0296] 9. The particle according to item 8, wherein the bisphosphonate is selected from etidronate, clodronate, tiludronate, pamidronate, neridronate, opaldronate, alendronate, ibandronate, risedronate, and zoledronic acid.

[0297] 10. The particle according to item 7, wherein the phosphonate is a polyphosphonate.

[0298] 11. The particles according to claim 10, wherein the polyphosphonate is selected from EDTMP-ethylenediaminetetra(methylenephosphonic acid), DOTMP-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacyl-tetra(methylphosphonic acid) and DTPMP-diethylenetriaminepenta(methylenephosphonic acid).

[0299] 12. The particle according to items 1-11, wherein the radioactive nuclide is selected from... 225 Ra、 224 Ra、 223 Ra、225 Ac、 227 Th、 211 At、 213 Bi、 212 Pb, 64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y、 220 Rn、 216 Po、 212 Po、 208 Tl、 18 F, 67 Ga、 86 Y、 99m Tc, 111 In、 203 Pb, 83 Sr、 152 Tb and 155 Tb.

[0300] 13. The particle according to items 1-12, wherein the radionuclide is selected from alpha-radionuclides suitable for treatment, which are derived from... 225 Ac、 211 At、 213 Bi、 212 Bi、 225 Ra、 224 Ra、 223 Ra and 227 Th is composed of.

[0301] 14. The particle according to items 1-12, wherein the radionuclide is selected from β-radionuclides suitable for treatment, which are derived from... 64 Cu、 67 Cu、 166 Ho、 177 Lu、 32 P, 186 Re、 188 Re、 153 Sm、 89 Sr、 161 Tb, 90 Y is composed of.

[0302] 15. The particle according to items 1-12, wherein the radionuclide is a β emitter with α-generation suitable for treatment, which is 212 Pb.

[0303] 16. The particle according to items 1-12, wherein the radioactive nuclide is selected from those emitting alpha. 224 Ra and progeny radionuclides 220 Rn、 216 Po、 212 Pb, 212 Bi、 212 Po and 208 Tl.

[0304] 17. The particle according to items 1-12, wherein the radionuclide is selected from radionuclides suitable for imaging, which are composed of 18 F, 67 Ga、 86 Y、 99m Tc, 111 In、 203 Pb, 83 Sr、 152 Tb and 155 Tb composition.

[0305] 18. The particle according to any one of items 1-17, wherein the particle size is from 1 nm to 500 μm.

[0306] 19. The particles according to any one of claims 1-18, wherein the degradable compound is selected from PEG-modified CaCO3, protein-modified CaCO3 including mAb and Fab, carbohydrate-modified CaCO3, lipid-modified CaCO3, vitamin-modified CaCO3, organic compound-modified CaCO3, polymer-modified CaCO3 and / or inorganic crystal-modified CaCO3.

[0307] 20. A composition comprising one or more particles as described in any one of items 1-19.

[0308] 21. A composition comprising one or more particles as described in any one of items 1-19 and a diluent, a carrier, a surfactant, and / or an excipient.

[0309] 22. The pharmaceutical composition according to claim 21 or the composition according to claim 15, which is prepared with a radionuclide amount of 1 kBq to 10 GBq per dose or with a radionuclide amount of 50 MBq to 100 GBq suitable for multi-dose industrial-scale production.

[0310] 23. The composition according to claim 20, or the pharmaceutical composition according to any one of claims 21-22, wherein the composition is a particle suspension comprising monodisperse or polydisperse particles as described in claims 1-19.

[0311] 24. The composition or pharmaceutical composition according to any one of items 20-23, which is suitable for parenteral use, such as for intravenous, intracavitary and / or intratumoral injection.

[0312] 25. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, used as a medicine.

[0313] 26. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, for use in endovascular treatment, radioembolization or radiotherapy for synovectomy.

[0314] 27. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, for the treatment of cancer.

[0315] 28. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, for use according to items 26-27, wherein the cancer is selected from intraperitoneal carcinoma, intracranial carcinoma, pleural carcinoma, bladder cancer, gastric cardia cancer, subarachnoid carcinoma, non-cavitary targets such as melanoma, and non-small cell lung cancer.

[0316] 29. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, for imaging.

[0317] 30. The particles according to any one of items 1-19 or the compositions or pharmaceutical compositions according to any one of items 20-24, for use as described in items 26-27, in combination with other cancer therapies, such as chemotherapy such as taxanes (e.g., paclitaxel, docetaxel), platinum (e.g., carboplatin, cisplatin), doxorubicin, mitomycin), DNA repair inhibitors such as PARP inhibitors (e.g., olaparib, rucaparib, niraparib, talapazobal, veliparib, pamiparib, CEP 9722, E7016 and 3-aminobenzamide), and radioimmunotherapy.

[0318] 31. The particles according to any one of items 1-19 or the composition or pharmaceutical composition according to any one of items 20-24, which is a medical device or is contained in a medical device.

[0319] 32. The composition or pharmaceutical composition according to items 20-24, wherein the concentration of the phosphonate and / or phosphate compound is from 1 microgram to 1000 milligrams per milliliter, for example from 0.1 milligrams to 10 milligrams per milliliter of final solution, or from 1 microgram to 1000 milligrams per gram of particles in the final solution.

[0320] 33. A method for preparing particles as described in any one of items 1-19, the method comprising contacting a degradable compound, a radionuclide, and a phosphorus-containing additive with or without the use of a radionuclide carrier.

[0321] 34. A method for preparing particles as described in item 33, wherein a degradable compound and a radionuclide form particles in an initial step, and the particles are subsequently coated with a phosphorus-containing additive.

[0322] 35. A composition or suspension comprising particles, wherein the particles comprise a degradable compound, a radionuclide, and a phosphorus-containing additive, and wherein the phosphorus-containing additive is associated with the particles by being present in the composition or suspension.

[0323] 36. The composition or suspension according to claim 35, wherein the phosphorus-containing additive is part of the particles.

[0324] 37. The composition or suspension according to items 35-36, wherein the phosphorus-containing additive is part of the composition or suspension of particles.

[0325] 38. The composition or suspension according to items 35-37, wherein the phosphorus-containing additive is part of the particles and also part of the composition or suspension of the particles.

[0326] 39. The composition or suspension according to items 35-38, wherein the particle suspension is a mixture of a solid phase and a liquid phase.

[0327] 40. The composition or suspension according to items 35-39, wherein the phosphorus-containing additive is in the liquid phase.

[0328] 41. The composition or suspension according to items 35-40, wherein the phosphorus-containing additive is a solid phase.

[0329] 42. The composition or suspension according to items 35-41, wherein the phosphorus-containing additive is in both a solid and a liquid phase.

[0330] 43. The composition or suspension according to items 35-42, wherein the phosphorus-containing additive is on the particle surface or embedded in the particles, or both are on the surface or embedded in the solid phase.

Claims

1. A particle comprising a degradable compound, a radionuclide and a phosphorous containing additive, characterized in that, - the degradable compound comprises CaC03and - the radionuclide is 224 Ra, and the phosphorus-containing additive is EDTMP-ethylenediaminetetra(methylene phosphonic acid), or the radionuclide is 212 Pb, and the phosphorus-containing additive is pamidronate.

2. The particle of claim 1, wherein the size of the particle is from 1 nm to 500 pm.

3. A composition comprising one or more particles according to any one of claims 1-2.

4. A pharmaceutical composition comprising the composition according to claim 3, a carrier, a surfactant and / or an excipient.

5. The composition according to claim 3 or the pharmaceutical composition according to claim 4, characterized in that, the phosphorous containing additive is associated with the particle by being present in the composition or suspension, by being part of the particle, on the surface of the particle, in the dispersion of the particle, as part of the particle composition or suspension and / or dispersion, or as part of the particle and as part of the particle composition or suspension.

6. The composition or pharmaceutical composition according to claim 5, characterized in that, the phosphorous containing additive is associated with the particle by 0.01-80% in or on the particle.

7. The composition according to claim 3 or the pharmaceutical composition according to claim 4, characterized in that, it is prepared with an amount of radionuclide per dose of 1 kBq to 10 GBq or with an amount of radionuclide of 50 MBq to 100 GBq suitable for multi-dose industrial scale production.

8. Use of a particle according to any one of claims 1-2 or a composition or pharmaceutical composition according to any one of claims 3-7 for the manufacture of a medicament for the treatment of cancer, wherein the cancer is selected from the group consisting of intraperitoneal cancer, intracranial cancer, pleural cancer, bladder cancer, cardia cancer, melanoma, non-small cell lung cancer.

9. The use according to claim 8, in combination with other cancer therapies, including: chemotherapy, DNA repair inhibitors, and radioimmunotherapy; the chemotherapy comprises taxanes, platinums, doxorubicin, mitomycin; the taxanes comprise paclitaxel, docetaxel; the platinums comprise carboplatin, cisplatin; the DNA repair inhibitors comprise PARP inhibitors; the PARP inhibitors comprise olaparib, rucaparib, niraparib, talazoparib, veliparib, pamiparib, CEP 9722, E7016 and 3-aminobenzamide.

10. A method of preparing the particle of any one of claims 1-2, characterized in that, the method comprises bringing the degradable compound, the radionuclide and the phosphorous containing additive in contact with each other with or without the use of a carrier for the radionuclide.

11. The method of claim 10, wherein, the degradable compound and the radionuclide form a particle in an initial step, followed by coating the particle with the phosphorous containing additive or at least partially associating the phosphorous containing additive with the particle.

12. The method of claim 11, wherein, the degradable compound and the radionuclide form a particle in an initial step, followed by incorporating the phosphorous containing additive in the particle. the degradable compound and the radionuclide form a particle in an initial step, followed by incorporating the phosphorous containing additive in the particle.

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