Formulation of Hydroxypyridonate Actinide / Lanthanide In Vitro Removal Agent

Abstract

To provide pharmaceutical formulations comprising a 1,2-HOPO chelating agent and / or 3,2-HOPO chelating agent.SOLUTION: Exposure to radionuclides accidentally or deliberately scattered by a radiological dispersion device or deposited from a nuclear power plant accident or nuclear device detonation could result in the contamination of a large population. As internalized radionuclides are highly toxic and may cause both acute and chronic radiation injury, such contamination event would have dramatic public health consequences. The present invention can solve this problem.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 384,087, filed September 6, 2016, which is hereby incorporated by reference in its entirety.

[0002] Statement of Government Support This invention was made possible under contract by the National Institute of Allergy and Infectious Diseases under Contract No. HHSN272201000046C and the Biomedical Advanced Research and Development Authority. This invention was made with Government support under U.S. Department of Energy Contract No. DE-AC02-05CH11231 under U.S. Department of Energy Contract No. IPIAA12OS99609. The Government has certain rights in this invention.

[0003] background FIELD OF THEINVENTION The present invention relates generally to formulations for the treatment of metal poisoning. [Background technology]

[0004] 2. Description of Related Art Exposure to radionuclides, either accidentally or intentionally dispersed by radioactive material dispersal devices or deposited by nuclear power plant accidents or nuclear device explosions, can result in contamination of large populations. Such contamination events could have dramatic public health consequences, since internalized radionuclides are highly toxic and can cause both acute and chronic radiation injury.

[0005] Extracorporeal removal by chelating agents is the only way to reduce exposure to certain incorporated isotopes, and diethylenetriaminepentaacetic acid (DTPA) has been the standard treatment for actinide / lanthanide extracorporeal removal since its development and use by the U.S. Atomic Energy Commission in the 1950s. Summary of the Invention Means for Solving the Problems

[0006] Gist of the Preferred Embodiment Embodiments herein provide a pharmaceutical composition comprising a 1,2-HOPO chelating agent in an amount of about 300 to about 1500 mg, and sodium oleate. In some embodiments, the 1,2-HOPO chelating agent is 3,4,3-LI-1,2-HOPO. In some embodiments, sodium oleate is present in an amount of about 70 to about 130 mg. In some embodiments, sodium oleate is present at 8 to 12% of the total weight of the composition. In some embodiments, sodium oleate is about 11% of the total weight of the composition. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 1500 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 400 to 1200 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 300 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 600 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 1500 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 400 to 1200 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 300 mg. In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 600 mg. In some embodiments, the pharmaceutical composition is packaged as a tablet. In some embodiments, the pharmaceutical composition is contained in a capsule. In some embodiments, the pharmaceutical composition is contained in one or more granules. In some embodiments, the pharmaceutical composition is packaged as a tablet. In some embodiments, the pharmaceutical composition is contained in a capsule. In some embodiments, the pharmaceutical composition is contained in one or more granules.

[0007] This is particularly useful when administered to a subject that has been exposed to, is in contact with, or is contaminated with one or more known or unknown actinides and / or lanthanides or mixtures thereof.

[0008] The foregoing aspects and other aspects can be easily understood by those skilled in the art from the following description of exemplary embodiments when read in conjunction with the accompanying drawings. In embodiments of the present invention, for example, the following items are provided. (Item 1) A pharmaceutical composition comprising a 1,2-HOPO chelating agent in an amount of about 300 to about 1500 mg and sodium oleate. (Item 2) The pharmaceutical composition according to Item 1, wherein the 1,2-HOPO chelating agent is 3,4,3-LI-1,2-HOPO. (Item 3) The pharmaceutical composition according to Item 2, wherein sodium oleate is present in an amount of about 70 to about 130 mg. (Item 4) The pharmaceutical composition according to Item 2, wherein sodium oleate is present in an amount of 8 to 12% of the total weight of the composition. (Item 5) The pharmaceutical composition according to Item 4, wherein sodium oleate is about 11% of the total weight of the composition. (Item 6) The pharmaceutical composition according to Item 4, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 1500 mg. (Item 7) The pharmaceutical composition according to Item 5, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 400 to 1200 mg. (Item 8) The pharmaceutical composition according to Item 5, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 300 mg. (Item 9) The pharmaceutical composition according to Item 5, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 600 mg. (Item 10) The pharmaceutical composition according to Item 2, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 1500 mg. (Item 11) The pharmaceutical composition according to item 2, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 400 to 1200 mg. (Item 12) The pharmaceutical composition according to item 2, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 100 to 300 mg. (Item 13) The pharmaceutical composition according to item 2, wherein the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of 600 mg. (Item 14) The pharmaceutical composition according to item 2, which is packaged as a tablet. (Item 15) The pharmaceutical composition according to item 2, which is contained in a capsule. (Item 16) The pharmaceutical composition according to item 2, which is contained in one or more granules. (Item 17) The pharmaceutical composition according to item 8 or 9, which is packaged as a tablet. (Item 18) The pharmaceutical composition according to item 8 or 9, which is contained in a capsule. (Item 19) The pharmaceutical composition according to item 8 or 9, which is contained in one or more granules.

Brief Description of the Drawings

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[0040] Figure 19A shows data related to the retention of radioactivity from 14 C]-3,4,3-LI(1,2-HOPO) in the liver and kidneys of male and female mice after intravenous administration.

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[0047] Figure 20A shows data related to the retention of radioactivity from 14 C]-3,4,3-LI(1,2-HOPO) in male and female rats after intravenous administration.

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[0052] Figure 21A shows the clearance of 3,4,3-LI(1,2-HOPO) in the plasma of male dogs.

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Mode for Carrying Out the Invention

[0055] Detailed Description of Various Embodiments Possible consequences of a large-scale radiological event include not only large-scale external radiation exposure of a population, but also uncontrolled dispersion of radionuclides and resulting internal contamination. When planning an emergency response to radiological and nuclear incidents, the need to treat contaminated individuals must be considered. In addition to meeting the desired criteria for post-exposure treatment, such as safety, ease of administration, and broad effectiveness against multiple radionuclides and levels of concern, an ideal countermeasure may include immediacy, induce minimal to no side effects that degrade performance, be compatible with current military countermeasures against chemical, biological, radiological, nuclear, and explosive agents, and require minimal logistical burden. Hydroxypyridinone-based actinide decorporation agents have been shown to be the most promising as decorporation strategies for various radionuclides of concern, including actinides, plutonium, and americium.

[0056] Various formulations for decorporation agents are provided herein.

[0057] The following disclosure provides a series of brief definitions, then provides further details regarding the various formulations of the chelators provided herein, and then provides a series of examples regarding various embodiments.

[0058] Definitions The term "emergency" includes the following: (a) An accidental release event of radioisotopes into the environment resulting from any nuclear accident. (b) Any accidental release of harmful nuclides into the environment. (c) Fallout, including that which occurs during the course of normal experimental, diagnostic, or therapeutic purposes. (d) Any kind of accidental uptake and retention of radionuclides by human or animal subjects. (e) Any other kind of exposure to volatile radionuclides. (f) Any kind of radiological accident.

[0059] The term "pharmaceutically acceptable salt", as used herein and in particular when referring to pharmaceutically acceptable salts of compounds comprising 3,4,3-LI(1,2-HOPO), refers to any pharmaceutically acceptable salt of the compound, preferably an acid addition salt of the compound.

[0060] The terms "pure", "purified", "substantially purified", and "isolated", as used herein, refer to a compound of an embodiment that, when found in its natural state, does not contain other non-similar compounds that would be expected to associate in its natural state. In certain embodiments described herein as "pure", "purified", "substantially purified", or "isolated", the compound can constitute at least 0.5% - 1%, 1% - 5%, 5% - 10%, 10% - 20%, 20% - 50%, 50% - 70%, 70% - 90%, 90% - 95%, 95% - 99%, and 99% - 100%. In some embodiments, the amount of the compound is at least 50% or 75% of the mass of a given sample by weight. "Functional purity" is a measure of the amount of a particular compound in a sample or product relative to other compounds in the sample that can adversely affect the function of that compound. Thus, other components in the sample (e.g., water) that do not interfere with the activity of the compound are not used in determining the purity of the sample or product.

[0061] The terms "derivative", "variant", or other similar terms refer to a compound that is an analog of another compound.

[0062] The term "and / or" indicates both the option of "and" as well as the option of "or" in that particular context. However, unless otherwise indicated herein, the use of the terms "or" or "and" encompasses the description of both options equally. Thus, the use of the term "or" is not taken to exclude the option of "and" unless additional context indicates otherwise (this definition does not apply to the language of the claims). The use of terms in the singular or plural encompasses both options (singular or plural), as well as combinations of both options (singular and plural), unless otherwise indicated.

[0063] As used herein, the term "inhibit" refers to any statistically significant decrease in the deleterious effects of a metal, including complete blockade of activity. For example, "inhibit" can refer to a decrease of about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% in the deleterious effects of a metal.

[0064] The term "patient" includes human and other mammalian subjects that receive either a prophylactic or therapeutic treatment.

[0065] The terms "treat" or "prevent" do not require complete treatment or complete prevention under all circumstances. Delaying the onset of a disorder or its symptoms, or a decrease in the number of symptoms, can be an appropriate "prevention" in some embodiments. Similarly, a decrease in the severity of the symptoms of a disorder can also be an effective treatment of the disorder. "Prophylactic treatment" refers to administration of a compound before exposure to a deleterious compound (e.g., a metal, such as plutonium or an MRI contrast agent). Treatment can also be in response to exposure, e.g., as a response therapy. Treatment also encompasses amelioration, extracorporeal removal, and / or decontamination.

[0066] "Therapeutically effective amount" means the amount of a chelating agent, such as 3,4,3-LI(1,2-HOPO), 5-LIO(Me-3,2-HOPO), and / or DTPA, that induces a biological or pharmaceutical response in a tissue system, animal, or human sought by a researcher, veterinarian, physician, or other clinician, and this response includes alleviation of the symptoms of the disease or disorder being treated. The specific amount of chelating agent required to induce a biological or pharmaceutical response is expected to depend on a number of factors including, but not limited to, the disease or disorder being treated, the chelating agent being administered, the method of administration, and the condition of the patient.

[0067] "Mammal" as used herein refers to any animal considered to be a mammal. Preferably, the mammal is a human.

[0068] The term "pharmaceutical agent or drug" as used herein refers to a chemical compound or composition that can induce a desired therapeutic effect when appropriately administered to a patient. Other chemical terms herein are used according to their conventional usage in the art as exemplified by The McGraw-Hill Dictionary of Chemical Terms (Parker, S., ed., McGraw-Hill, San Francisco (1985)), which is incorporated herein by reference.

[0069] The term "heavy metal" refers to one or more of the transition metals, metalloids, metallic elements of groups 13, 14, and 15 of the periodic table, actinides, or lanthanides. Examples of heavy metals include, for example, gadolinium, lead, tin, cadmium, yttrium, scandium, and plutonium. Pharmaceutical formulation

[0070] In some embodiments, the pharmaceutical composition formulation comprises a chelating agent and one or more additional components. In some embodiments, the chelating agent is a 1,2-HOPO chelating agent. In some embodiments, the chelating agent is 3,4,3-LI-1,2-HOPO.

[0071] In some embodiments, the pharmaceutical composition comprises a 1,2-HOPO chelating agent in an amount of about 300 to about 1500 mg, and sodium oleate. In some embodiments, the 1,2-HOPO chelating agent of the pharmaceutical composition is 3,4,3-LI-1,2-HOPO.

[0072] In some embodiments, any amount of sodium oleate appropriate for the intended use can be used. In some embodiments, the amount of sodium oleate present is about 50 to about 150 mg, such as about 70 to about 130 mg. In some embodiments, sodium oleate is present in an amount of about 5 to about 20% of the total weight of the composition, such as about 8 to 12% of the total weight of the composition or about 11% of the total weight of the composition. Other amounts described herein are also applicable to various uses.

[0073] In some embodiments, the 3,4,3-LI-1,2-HOPO chelating agent is present in an amount of about 50 to about 2000 mg, such as about 100 to 1500 mg, about 400 to 1200 mg, about 100 to 300 mg, or about 600 mg. Other amounts described herein are also applicable to various uses.

[0074] The amount of chelating agent that can be combined with a pharmaceutically acceptable carrier to produce a single dosage form can vary depending on the subject being treated and the particular mode of administration. Suitable dosage levels of the chelating agent include from about 1 mg to about 500 mg per kg of body weight per day. In some embodiments, suitable dosage levels are from about 20 mg to about 100 mg per kg of body weight per day. In some embodiments, suitable dosage levels for 3,4,3-LI-1,2-HOPO are from about 10 μmol to about 100 μmol per kg of body weight. In some embodiments, suitable dosage levels for 5-LIO-Me-3,2-HOPO are from about 30 μmol to about 200 μmol per kg of body weight. Dosage unit forms are typically expected to contain from about 20 mg to about 100 mg of the chelating agent. Additionally, the pharmaceutical composition can be administered intermittently, i.e., at intervals of daily, twice a week, or once a week. However, it is expected that the specific dosage level for a particular subject will depend on a variety of factors. These factors include the activity of the specific compound used, the age, weight, general health, sex, and diet of the subject, the time and route of administration of the chelating agent and the rate of excretion, the combination of chelating agents used in the treatment, and the severity of the particular disease or condition for which treatment is sought.

[0075] In some embodiments, the pharmaceutical composition is packaged as tablets, in capsules, and / or in one or more granules.

[0076] Suitable modes of administration of the pharmaceutical composition include, but are not limited to, oral, topical, aerosol, inhalation by spray, parenteral, subcutaneous, intravenous, intramuscular, intraperitoneal, rectal, and vaginal administration. As used herein, the term parenteral includes subcutaneous injection, as well as intravenous, intrathecal, intramuscular, and intracardiac injection or infusion techniques. The particular mode of administration is one that delivers the compounds of the invention to the actual or potential site of radionuclide contamination in the subject. The pharmaceutical composition can be in solid, semi-solid, and / or liquid form. In some embodiments, any of the above-described formulations can be used with any of the metals provided herein.

[0077] In some embodiments, the formulation can include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers described herein, such as vehicles, adjuvants, excipients, and diluents, are well known to those skilled in the art and are readily available. In some embodiments, the carrier is chemically inert to the compounds of the invention and has no adverse side effects or toxicity under the conditions of use. In some embodiments, the pharmaceutically acceptable carrier is pyrogen-free. Pharmaceutically acceptable carriers that can be used include, but are not limited to, water, glucose, lactose, acacia gum, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, and urea.

[0078] Suitable pharmaceutical compositions for oral administration include, but are not limited to, (a) liquid preparations, (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient as a solid or granule, (c) powders, (d) suspensions, and (e) suitable emulsions. Liquid preparations may contain diluents, such as water and alcohol, and optionally may contain pharmaceutically acceptable surfactants. Capsule forms can be of the gelatin type with a normal hard or soft shell, containing, for example, surfactants, lubricants, and inert fillers. Tablet forms may contain one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, etc. Tablets may further contain one or more coloring agents, diluents, buffering agents, disintegrants, wetting agents, preservatives, or flavoring agents.

[0079] The pharmaceutical composition can be made into an aerosol preparation for administration through inhalation, either alone or in combination with other suitable components. These aerosol preparations can be placed in a pressurized acceptable propellant (such as dichlorodifluoromethane, propane, nitrogen, etc.) or a non-pressurized preparation (such as a nebulizer or atomizer). When the infected site of the subject is the lung, the preferred mode of administration is inhalation of the aerosol preparation either orally or nasally. In particular, the aerosol preparation can contain respirable-sized particles having an average particle size of 5 μm to 500 μm, but is not limited thereto.

[0080] The pharmaceutical composition may be an injectable preparation. The requirements for an effective carrier in an injectable composition are well-known to those skilled in the art (see, for example, Pharmaceutics and Pharmacy Practice, J. B. Lippincott Company, Philadelphia, Pa., edited by Banker and Chalmers, pages 238-250 (1982) and ASHP Handbook on Injectable Drugs, Toissel, 4th edition, pages 622-630 (1986)). In certain embodiments, the injectable composition is administered intravenously. Formulations suitable for parenteral administration may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous isotonic sterile injectable solutions, and aqueous and non-aqueous sterile suspensions that may contain suspending agents, solubilizing agents, thickening agents, stabilizers, and preservatives.

[0081] The pharmaceutical composition may further comprise an excipient. Excipients that may be used include one or more carriers, surfactants, thickening or emulsifying agents, solid binders, dispersion and suspension aids, solubilizing agents, coloring agents, flavoring agents, coating agents, disintegrating agents, lubricants, sweetening agents, preservatives, isotonic agents, and combinations thereof. The selection and use of suitable excipients are taught in Gennaro, Remington: The Science and Practice of Pharmacy, 20th edition (Lippincott Williams & Wilkins, 2003), the disclosure of which is incorporated herein by reference.

[0082] In some embodiments, the pharmaceutical composition may comprise one or more of the formulations of Table 1 and / or Table 2 below.

Table 1A

Table 1B

Table 2A

Table 2B

[0083] In some embodiments, the amount of the active ingredient can be maintained at any ratio provided in any formulation while increasing or decreasing.

[0084] In some embodiments, the formulation can comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more excipients. In some embodiments, pharmaceutically suitable excipients include, but are not limited to, mannitol, lactose monohydrate, compressed sugar, microcrystalline cellulose, hypromellose, povidone, pregelatinized starch, croscarmellose sodium, sodium starch glycolate, crospovidone, colloidal silicon dioxide, magnesium stearate, hydrogenated vegetable oil (type 1), and polysorbate 80.

[0085] In some embodiments, various forms of formulations can be used, including (i) powders, (ii) orally dispersible / soluble granules, (iii) chewable tablets, and / or (iv) conventional immediate-release tablets. Based on the studies conducted in the examples, nine formulation prototypes were shown to exhibit immediate drug release behavior and desired physical properties, and these were selected for API verification, gastric dissolution, and testing for related substances by a specified liquid chromatography method. Of these selected compositions, two are powder formulations in bottles, two are granule formulations, three are chewable tablet formulations, and two are conventional tablet formulations. The respective compositions of these formulations are summarized in the examples and Tables 1 and 2 above. All assays confirmed that these prototypes are suitable for further development.

[0086] In some embodiments, the powder formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, and optionally a mixture of microcrystalline cellulose and carboxymethyl cellulose. In some embodiments, the composition may comprise 1 g of 3,4,3-LI(1,2-HOPO) and 0.092 g of sodium oleate. In some embodiments, the composition may comprise 1 g of 3,4,3-LI(1,2-HOPO), 0.092 g of sodium oleate, and 1 g of a mixture of microcrystalline cellulose and carboxymethyl cellulose. In some embodiments, the composition may comprise 1 g of 3,4,3-LI(1,2-HOPO), 0.092 g of sodium oleate, and from 0 g to 1 g of a mixture of microcrystalline cellulose and carboxymethyl cellulose. Weights are listed in grams per unit of formulation.

[0087] In some embodiments, the formulation can be an orally dispersible / soluble granule formulation. This can include 1 g of 3,4,3-LI(1,2-HOPO), 0.092 g of sodium oleate, 0.075 g of croscarmellose sodium, and 1.833 g of microcrystalline cellulose and guar gum (by weight of the components per unit of the formulation). In some embodiments, the formulation can include 1 g of 3,4,3-LI(1,2-HOPO), 0.092 g of sodium oleate, 0.075 g of croscarmellose sodium, 1.533 mg / ml of lactose monohydrate, and 0.3 g of hypromellose.

[0088] In some embodiments, the orally dispersible / soluble granule formulation can include a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, croscarmellose sodium, a mixture of microcrystalline cellulose and guar gum, lactose monohydrate, and hypromellose. In some embodiments, the composition can include 1 g of 3,4,3-LI(1,2-HOPO), 0.092 g of sodium oleate, 0.075 g of croscarmellose sodium, 0 - 1.833 g of microcrystalline cellulose and guar gum, 0 - 1.533 mg / ml of lactose monohydrate, and 0 - 0.3 g of hypromellose. All weights are listed in grams per unit of the formulation unless otherwise indicated.

[0089] In some embodiments, the formulation can be a chewable tablet formulation. In some embodiments, the chewable tablet formulation can include a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, croscarmellose sodium, a mixture of microcrystalline cellulose and guar gum, magnesium stearate, lactose monohydrate, povidone, and crospovidone, and mannitol. In some embodiments, the composition can include 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0.075 g of croscarmellose sodium, 1.854 g of microcrystalline cellulose and guar gum, and 0.025 g of magnesium stearate. In some embodiments, the composition can include 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 1.929 g of lactose monohydrate, povidone, and crospovidone, and 0.025 g of magnesium stearate. In some embodiments, the composition can include 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0.075 g of croscarmellose sodium, 0.927 g of microcrystalline cellulose and guar gum, 0.9227 g of mannitol, and 0.025 g of magnesium stearate. In some embodiments, the composition can include 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0 to 0.075 g of croscarmellose sodium, 0 to 1.854 g of microcrystalline cellulose and guar gum, 0.025 g of magnesium stearate, 0 to 1.929 g of lactose monohydrate, povidone, and crospovidone, and 0 to 0.9227 g of mannitol. All weights are listed in grams per unit of the formulation.

[0090] In some embodiments, the immediate-release tablet formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, croscarmellose sodium, microcrystalline cellulose, colloidal silicon dioxide, and magnesium stearate. In some embodiments, the composition may comprise 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0.084 g of croscarmellose sodium, 0.41 g of microcrystalline cellulose, 0.005 g of colloidal silicon dioxide, and 0.005 g of magnesium stearate. In some embodiments, the composition may comprise 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0.092 g of croscarmellose sodium, 0.501 g of microcrystalline cellulose, 0.006 g of colloidal silicon dioxide, and 0.006 g of magnesium stearate. In some embodiments, the composition may comprise 0.5 g of 3,4,3-LI(1,2-HOPO), 0.046 g of sodium oleate, 0.084 - 0.092 g of croscarmellose sodium, 0.41 g - 0.501 g of microcrystalline cellulose, 0.005 g - 0.006 g of colloidal silicon dioxide, and 0.005 g - 0.006 g of magnesium stearate. All weights are listed in grams per unit of the formulation.

[0091] In some embodiments, the formulation may be stable for 6 months or longer under storage conditions of 25°C / 60% relative humidity and 40°C / 75% relative humidity. Such stable formulations can be powder formulations, chewable tablet formulations, immediate-release tablet formulations, 500 mg capsule formulations, and 100 mg capsule formulations. The respective compositions of these prototype formulations are summarized below.

[0092] In some embodiments, the powder formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, and a mixture of microcrystalline cellulose and carboxymethyl cellulose. In some embodiments, the composition may comprise 500 mg of 3,4,3-LI(1,2-HOPO), 46 mg of sodium oleate, and 500 mg of microcrystalline cellulose and carboxymethyl cellulose. All weights are listed in milligrams per unit of the formulation.

[0093] In some embodiments, the chewable tablet formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, croscarmellose sodium, a mixture of microcrystalline cellulose and guar gum, mannitol, and magnesium stearate. In some embodiments, the composition may comprise 500 mg of 3,4,3-LI(1,2-HOPO), 46 mg of sodium oleate, 75 mg of croscarmellose sodium, 927 mg of microcrystalline cellulose and guar gum, 927 mg of mannitol, and 25 mg of magnesium stearate. All weights are listed in milligrams per unit of the formulation.

[0094] In some embodiments, the immediate release tablet formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO), sodium oleate, croscarmellose sodium, microcrystalline cellulose, colloidal silicon dioxide, and magnesium stearate. In some embodiments, the composition may comprise 500 mg of 3,4,3-LI(1,2-HOPO), 46 mg of sodium oleate, 92 mg of croscarmellose sodium, 501 mg of microcrystalline cellulose, 6 mg of colloidal silicon dioxide, and 6 mg of magnesium stearate. The weights are listed in milligrams per unit of the formulation.

[0095] In some embodiments, a 500 mg capsule formulation comprising a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and sodium oleate can be provided. In some embodiments, the composition can comprise 500 mg of 3,4,3-LI(1,2-HOPO) and 55.6 mg of sodium oleate.

[0096] In some embodiments, a 100 mg capsule formulation comprising a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and sodium oleate can be provided. In some embodiments, the composition can comprise 100 mg of 3,4,3-LI(1,2-HOPO) and 11.1 mg of sodium oleate.

[0097] The formulations can be configured for administration by injection (ip) or orally (po) via continuous injection or gavage at various doses, including but not limited to human doses of 2.5, 12.5, 25, and 50 μmol / kg, using an acceptable conversion system from mouse doses based on body surface area to human equivalent doses (HED). Parenteral formulations can be pure 3,4,3-LI(1,2-HOPO), while oral formulations can comprise 3,4,3-LI(1,2-HOPO) and sodium oleate in a 90:10 weight ratio.

[0098] In some embodiments, a formulation for intraperitoneal injection can comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO). In some embodiments, the composition can comprise 3,4,3-LI(1,2-HOPO) at a concentration of 30 μmol / kg intraperitoneally (approximately equivalent to a human dose of 2.5 μmol / kg).

[0099] In some embodiments, the oral formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and sodium oleate. In some embodiments, the composition may comprise 3,4,3-LI(1,2-HOPO) and sodium oleate in a weight ratio of 90:10. In some embodiments, the composition may comprise 3,4,3-LI(1,2-HOPO) at an oral concentration of 150 μmol / kg. In some embodiments, the composition may comprise 3,4,3-LI(1,2-HOPO) at an oral concentration of 300 μmol / kg. In some embodiments, the composition may comprise 3,4,3-LI(1,2-HOPO) at an oral concentration of 300 μmol / kg. In some embodiments, the composition may comprise 3,4,3-LI(1,2-HOPO) at an oral concentration of 150 μmol / kg to 600 μmol / kg.

[0100] In some embodiments, the formulation for intraperitoneal injection, oral, or intravenous injection may comprise a pharmaceutically suitable composition of [14C]-3,4,3-LI(1,2-HOPO) and sodium oleate. In some embodiments, the composition may comprise [14C]-3,4,3-LI(1,2-HOPO) at a dosage of 1 μmol / kg and 0% sodium oleate. In some embodiments, the composition may comprise [14C]-3,4,3-LI(1,2-HOPO) at a dosage of 1 μmol / kg and 10% sodium oleate. In some embodiments, the composition may comprise [14C]-3,4,3-LI(1,2-HOPO) at a dosage of 1800 μmol / kg and 0% sodium oleate. In some embodiments, the composition may comprise [14C]-3,4,3-LI(1,2-HOPO) at a dosage of 1800 μmol / kg and 10% sodium oleate. In some embodiments, the composition may comprise [14C]-3,4,3-LI(1,2-HOPO) at a dosage of 1 μmol / kg to 1800 μmol / kg and 0 to 10% sodium oleate.

[0101] In some embodiments, an orally administered capsule formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO). In some embodiments, the composition may comprise 50 μmol / kg of 3,4,3-LI(1,2-HOPO). In some embodiments, the composition may comprise 100 μmol / kg of 3,4,3-LI(1,2-HOPO). In some embodiments, the composition may comprise 200 μmol / kg of 3,4,3-LI(1,2-HOPO). In some embodiments, the composition may comprise 50 μmol / kg to 200 μmol / kg of 3,4,3-LI(1,2-HOPO).

[0102] As outlined in the Examples (Examples 10-11), the enhancement of the permeability of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO) using an oral permeation enhancer was evaluated. Fifteen different permeation enhancers were evaluated for their ability to increase the permeability of 3,4,3-LI(1,2-HOPO) using an in vitro PAMPA assay with an artificial GIT lipid membrane. A significant increase in permeability was observed for one formulation containing 10 mg / mL of polysorbate 80 and 1 mg / mL of API. All other tested formulations showed no or little improvement in permeability.

[0103] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more excipients. In some embodiments, excipients include sodium lauryl sulfate, caprolactam, polysorbate 80, sodium deoxycholate, isopropyl myristate, 1-phenylpiperazine, piperine, menthone, Labrafil lipophilic WL 1349, Gelucire 44 / 14, Labrafil M2130 Examples include, but are not limited to, CS, Lubrafill M2125 CS, Mycetin 35-1, Pesiol, Labrasol, Sodium decyl sulfate, Sodium octyl sulfate, Decyl trimethyl ammonium bromide, Span-80 (sorbitan monooleate), Triton X-100, Sodium glycolate hydrate, Cholic acid, Heptanoic acid, Isopropyl palmitate, Methyl laurate, Sodium oleate, Urea, 1-octyl-2-pyrrolidone, 1-methylpiperazine, 1-methyl-2-pyrrolidinone, n-caproic acid, Sodium salicylate, (±)-limonene, L-fenchone, Cineole, Pinene oxide, 2-octyl-1-dodecanol, Cumin seed oil, Caproyl PGMC, Caproyl 90 (propylene glycol dicaprylate), Lauroglycol FCC, Lauroglycol 90, Labrafac PG, Transcutol, Gelsiol 50 / 13, and Lubrafill M1944 CS.

[0104] In some embodiments, the composition can include 3,4,3-LI(1,2-HOPO) at a concentration of 1 mg / ml and polysorbate 80 at a concentration of 10 mg / ml. In some embodiments, the composition can include 3,4,3-LI(1,2-HOPO) at a concentration of 1 mg / ml and 2-octyl-1-dodecanol at a concentration of 2.5 mg / ml. In some embodiments, the composition can include 3,4,3-LI(1,2-HOPO) at a concentration of 1 mg / ml and sodium oleate at a concentration of 2.5 mg / ml.

[0105] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more excipients. In some embodiments, pharmaceutically suitable excipients include sodium oleate, sodium lauryl sulfate, caprolactam, polysorbate 80, sodium deoxycholate, isopropyl myristate, 1-phenylpiperazine, piperine, menthone, Labrafac lipophile, Gelsiol 44 / 14, Labrafil M2130 CS, Labrafil M2125 CS, Maisine 35-1, Peceol, Labrasol, sodium decyl sulfate, sodium octyl sulfate, decyltrimethylammonium bromide, Span-80 (sorbitan monooleate), triton X-100, sodium glycolate hydrate, cholic acid, heptanoic acid, isopropyl palmitate, methyl laurate, sodium oleate, urea, 1-octyl-2-pyrrolidone, 1-methylpiperazine, 1-methyl-2-pyrrolidinone, n-caproic acid, sodium salicylate, (±)-limonene, L-fenchone, cineole, pinene oxide, 2-octyl-1-dodecanol, cumin seed oil, caproyl PGMC, caproyl 90 (propylene glycol dicaprylate), Lauroglycol FCC, Lauroglycol 90, Labrafac PG, Transcutol, Gelsiol 50 / 13, Labrafil M1944 CS, mannitol, compressible sugar, coprocessed microcrystalline cellulose of microcrystalline cellulose and guar gum and guar gum), coprocessed lactose monohydrate and povidone, microcrystalline cellulose, lactose monohydrate, povidone, HPMC, hypromellose, pregelatinized starch, croscarmellose sodium, sodium starch glycolate, crospovidone, colloidal silicon dioxide, magnesium stearate, microcrystalline cellulose and carboxymethyl cellulose, hydrogenated vegetable oil type 1, lactose monohydrate, povidone, and co-processed lactose monohydrate, povidone and crospovidone, co-processed microcrystalline cellulose and carboxymethyl cellulose, maltodextrin, sodium citrate, and / or sodium chloride, but are not limited thereto.

[0106] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more penetration enhancers. In some embodiments, penetration enhancers include, but are not limited to, sodium oleate, sodium lauryl sulfate, caprolactam, polysorbate 80, sodium deoxycholate, isopropyl myristate, 1-phenylpiperazine, piperine, menthone, Labrafac lipophile, Gelsiol 44 / 14, Labrafil M2130 CS, Labrafil M2125 CS, Maisine 35-1, Peceol, Labrasol, sodium decyl sulfate, sodium octyl sulfate, decyltrimethylammonium bromide, Span-80 (sorbitan monooleate), triton X-100, sodium glycolate hydrate, cholate, heptanoic acid, isopropyl palmitate, methyl laurate, sodium oleate, urea, 1-octyl-2-pyrrolidone, 1-methylpiperazine, 1-methyl-2-pyrrolidinone, n-caproic acid, sodium salicylate, (±)-limonene, L-fenchone, cineole, pinene oxide, 2-octyl-1-dodecanol, cumin seed oil, caproyl PGMC, caproyl 90 (propylene glycol dicaprylate), lauroglycol FCC, lauroglycol 90, Labrafac PG, Transcutol, Gelsiol 50 / 13, and / or Labrafil M1944 CS.

[0107] In some embodiments, the composition may comprise 1 wt% to 10 wt% of sodium oleate. In some embodiments, the composition may comprise 5 mg to 100 mg of sodium oleate.

[0108] In some embodiments, the composition may comprise sodium lauryl sulfate at a concentration of 0.1 mg / ml. In some embodiments, the composition may comprise caprolactam at a concentration of 2.5 mg / ml.

[0109] In some embodiments, the composition may contain polysorbate 80 at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain polysorbate 80 at a concentration of 2 mg / ml. In some embodiments, the composition may contain polysorbate 80 at a concentration of 10 mg / ml. In some embodiments, the composition may contain polysorbate 80 at a concentration of 2 mg / ml to 10 mg / ml.

[0110] In some embodiments, the composition may contain sodium deoxycholate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain sodium deoxycholate at a concentration of 10 mg / ml. In some embodiments, the composition may contain sodium deoxycholate at a concentration of 2.5 mg / ml to 10 mg / ml.

[0111] In some embodiments, the composition may contain isopropyl myristate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain isopropyl myristate at a concentration of 10 mg / ml. In some embodiments, the composition may contain isopropyl myristate at a concentration of 2.5 mg / ml to 10 mg / ml.

[0112] In some embodiments, the composition may contain 1-phenylpiperazine at a concentration of 2.5 mg / ml.

[0113] In some embodiments, the composition may contain piperine at a concentration of 2.5 mg / ml.

[0114] In some embodiments, the composition may contain menthone at a concentration of 2.5 mg / ml.

[0115] In some embodiments, the composition may contain Lubrafac lipophilic WL 1349 at a concentration of 5 mg / ml.

[0116] In some embodiments, the composition may contain Gelsia 44 / 14 at a concentration of 5 mg / ml. In some embodiments, the composition may contain Gelsia 44 / 14 at a concentration of 20 mg / ml. In some embodiments, the composition may contain Gelsia 44 / 14 at a concentration of 1 mg / ml to 40 mg / ml.

[0117] In some embodiments, the composition may contain Lubrafill M2130 CS at a concentration of 5 mg / ml. In some embodiments, the composition may contain Lubrafill M2125 CS at a concentration of 5 mg / ml.

[0118] In some embodiments, the composition may contain Mycicin 35-1 at a concentration of 5 mg / ml. In some embodiments, the composition may contain Mycicin 35-1 at a concentration of 20 mg / ml. In some embodiments, the composition may contain Mycicin 35-1 at a concentration of 1 mg / ml to 40 mg / ml.

[0119] In some embodiments, the composition may contain Peseol 35-1 at a concentration of 5 mg / ml. In some embodiments, the composition may contain Peseol 35-1 at a concentration of 20 mg / ml. In some embodiments, the composition may contain Peseol 35-1 at a concentration of 1 mg / ml to 40 mg / ml.

[0120] In some embodiments, the composition may contain lovastatin at a concentration of 5 mg / ml. In some embodiments, the composition may contain sodium decyl sulfate at a concentration of 0.2 mg / ml. In some embodiments, the composition may contain sodium octyl sulfate at a concentration of 0.2 mg / ml. In some embodiments, the composition may contain decyltrimethylammonium bromide at a concentration of 1 mg / ml. In some embodiments, the composition may contain span-80 (sorbitan monooleate) at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain triton X-100 at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain sodium glycolate hydrate at a concentration of 1.0 mg / ml. In some embodiments, the composition may contain cholic acid at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain heptanoic acid at a concentration of 2.5 mg / ml.

[0121] In some embodiments, the composition may contain isopropyl palmitate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain methyl laurate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain sodium oleate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain urea at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain 1-octyl-2-pyrrolidone at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain 1-methylpiperazine at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain 1-methyl-2-pyrrolidinone at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain n-caproic acid at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain sodium salicylate at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain (±)-limonene at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain L-fenchone at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain cineole at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain pinene oxide at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain 2-octyl-1-dodecanol at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain cumin seed oil at a concentration of 2.5 mg / ml. In some embodiments, the composition may contain caproyl PGMC at a concentration of 5 mg / ml. In some embodiments, the composition may contain caproyl 90 (propylene glycol dicaprylate) at a concentration of 5 mg / ml.

[0122] In some embodiments, the composition may contain laureth glycol FCC at a concentration of 5 mg / ml. In some embodiments, the composition may contain laureth glycol 90 at a concentration of 5 mg / ml. In some embodiments, the composition may contain Labrafac PG at a concentration of 5 mg / ml. In some embodiments, the composition may contain transcutol at a concentration of 5 mg / ml. In some embodiments, the composition may contain Gelucire 50 / 13 at a concentration of 5 mg / ml. In some embodiments, the composition may contain Labrafil M1944 CS at a concentration of 5 mg / ml.

[0123] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more diluents. In some embodiments, diluents include, but are not limited to, mannitol, compressible sugar, co-processed microcrystalline cellulose and guar gum, co-processed lactose monohydrate and povidone, microcrystalline cellulose, and lactose monohydrate. In some embodiments, the composition may contain 10 wt% diluent. In some embodiments, the composition may contain 70 wt% diluent. In some embodiments, the composition may contain 10 wt% to 70 wt% diluent. In some embodiments, the composition may contain mannitol at a concentration of 2 mg / ml. In some embodiments, the composition may contain compressible sugar at a concentration of 2 mg / ml.

[0124] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more binders. In some embodiments, binders include, but are not limited to, povidone, HPMC, hypromellose, and pregelatinized starch. In some embodiments, the composition may contain 10 wt% binder. In some embodiments, the composition may contain 70 wt% binder. In some embodiments, the composition may contain 10 wt% to 70 wt% binder.

[0125] In some embodiments, the composition may contain povidone at a concentration of 2 mg / ml. In some embodiments, the composition may contain hypromellose at a concentration of 2 mg / ml. In some embodiments, the composition may contain pregelatinized starch at a concentration of 2 mg / ml.

[0126] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more disintegrants. In some embodiments, disintegrants include, but are not limited to, croscarmellose sodium, sodium starch glycolate, and crospovidone. In some embodiments, the composition may contain 2 wt% of the disintegrant. In some embodiments, the composition may contain 8 wt% of the disintegrant. In some embodiments, the composition may contain 2 wt% - 8 wt% of the disintegrant.

[0127] In some embodiments, the composition may contain croscarmellose sodium at a concentration of 2 mg / ml. In some embodiments, the composition may contain sodium starch glycolate at a concentration of 2 mg / ml. In some embodiments, the composition may contain crospovidone at a concentration of 2 mg / ml.

[0128] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more lubricants and glidants. In some embodiments, lubricants and glidants include, but are not limited to, colloidal silicon dioxide and magnesium stearate. In some embodiments, the composition may contain 0.2 wt% of the lubricant and glidant. In some embodiments, the composition may contain 20 wt% of the lubricant and glidant. In some embodiments, the composition may contain 0.2 wt% - 20 wt% of the lubricant and glidant.

[0129] In some embodiments, the composition may contain colloidal silicon dioxide at a concentration of 2 mg / ml. In some embodiments, the composition may contain magnesium stearate at a concentration of 2 mg / ml.

[0130] In some embodiments, the formulation may comprise a pharmaceutically suitable composition of 3,4,3-LI(1,2-HOPO) and one or more other excipients. In some embodiments, other excipients include, but are not limited to, microcrystalline cellulose and carboxymethyl cellulose, microcrystalline cellulose and guar gum, hydrogenated vegetable oil type 1, lactose monohydrate, povidone, and co-processed cross-povidone, co-processed microcrystalline cellulose and carboxymethyl cellulose, maltodextrin, sodium citrate, and sodium chloride.

[0131] In some embodiments, the composition may comprise hydrogenated vegetable oil type 1 at a concentration of 2 mg / ml. In some embodiments, the composition may comprise sodium citrate at a concentration of 0.008 M. In some embodiments, the composition may comprise sodium chloride at a concentration of 0.14 M.

[0132] Suitable 1,2-HOPO and 3,2-HOPO chelating agents for use in the present invention are taught in U.S. Patent Nos. 4,698,431 ("Hydroxypyridonate Chelating Agents"), 5,634,901 ("3-Hydroxy-2(1H)-pyridonate Chelating Agents"), and 5,892,029 ("3-Hydroxy-2(1H)-pyridonate Chelating Agents"), all of which are incorporated herein by reference.

[0133] Suitable 1,2-HOPO chelating agents include, but are not limited to, molecules defined by the following structure

Chemical formula

Chemical formula

[0134] Suitable 1,2-HOPO chelating agents include

Chemical formula

Chemical formula

[0135] Suitable 1,2-HOPO and 3,2-HOPO chelating agents include chelating agents composed of a plurality of chelating functional units linked by one or more linking members, including but not limited to those in which the chelating functional units are independently selected from the group consisting of

Chemical formula

Chemical formula

[0136] Suitable 3,2-HOPO chelating agents include the structure:

Chemical formula

[0137] Suitable 1,2-HOPO and suitable 3,2-HOPO are shown in Figure 1.

[0138] Methods for synthesizing 1,2-HOPO and 3,2-HOPO chelating agents are taught in U.S. Patent Nos. 4,698,431, 5,634,901, and 5,892,029, all of which are incorporated herein by reference.

[0139] The chelating agent can bind to or chelate cations of actinides and / or lanthanides, such as Eu, Pu, Np, Th, Am, and / or Cf, such as 152 Eu(III), 241 Am(III), 238 Pu(IV), 237 Np(IV), 237 Np(V), and 233 U(VI), or form a stable complex with it.

[0140] The embodiments provided herein include prodrugs of chelators. Such prodrugs are generally functional derivatives of compounds that are readily convertible in vivo to the required compound. Thus, in the present method, the term "administering" refers to a compound that, although not specifically disclosed, may be converted in vivo to the compound shown after administration to a subject in need thereof, and is used in the treatment of the various disorders described. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in Wermuth, "Designing Prodrugs and Bioprecursors" (Academic Press, 2003), pp. 561-586 in Wermuth, The Practice of Medicinal Chemistry, 2nd Edition. Prodrugs include esters that are hydrolyzed in vivo (e.g., in the human body) to produce the compounds of the present invention or salts thereof. Suitable ester groups include, without limitation, pharmaceutically acceptable aliphatic carboxylic acids, particularly those derived from alkanoic acids, alkenoic acids, cycloalkanoic acids, and alkanedioic acids, each having an alkyl or alkenyl moiety preferably having up to 6 carbon atoms. Exemplary esters include formate esters, acetate esters, propionate esters, butyrate esters, acrylate esters, citrate esters, succinate esters, and ethyl succinate esters.

[0141] Method of Use In some embodiments, a method for treating a subject for heavy metal exposure is provided. The method includes administering to a subject having an excess amount of one or more of heavy metals, actinides, and / or lanthanides, or mixtures thereof, a pharmaceutical formulation comprising a therapeutically effective amount of a 1,2-HOPO chelating agent. Further options for treatment are also provided in U.S. Patent Publication No. 20120214843, which is hereby incorporated by reference in its entirety. The method of treatment can include treating a subject in need thereof by administering to the subject in need thereof a therapeutically effective amount of one or more pharmaceutical compositions comprising a chelating agent (such as provided herein). In some embodiments, the subject has been exposed to, is in contact with, or is contaminated by one or more known or unknown actinides and / or lanthanides, or mixtures thereof.

[0142] It is to be understood that the invention is not limited to the specific embodiments described, and thus can, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting, as the scope of the invention is defined only by the appended claims.

[0143] When a range of values is provided, unless otherwise clearly indicated by the context, each value intermediate between the upper and lower limits of that range, down to one-tenth of the unit of the lower limit, is also to be understood as specifically disclosed. Each smaller range between any of the recited values or the values intermediate between any recited value and any other recited value is also included within the invention. The upper and lower limits of these smaller ranges may independently be included in or excluded from the range, and where either the upper or lower limit of the smaller range is included, where neither is included, or where both are included, each such range is also included within the invention, and there may be cases where any upper or lower limit in the recited range is specifically excluded. When the recited range includes one or both of the upper and lower limits, ranges excluding either or both of these included upper and lower limits are also included within the invention.

[0144] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described herein. All publications mentioned herein are incorporated by reference into this disclosure, disclosing and describing the methods and / or materials related to those cited in the publications.

[0145] It should be noted that as used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a chelating agent" includes a plurality of such chelating agents and the like.

[0146] Although the present invention has been described, the following examples are provided by way of illustration and not by way of limitation to exemplify the invention of the subject matter.

Example

[0147] (Example 1) Study on the Compatibility of 3,4,3-LI(1,2-HOPO) with Excipients Overview The goal of the analytical study described in this report was to evaluate the interaction between 3,4,3-LI(1,2-HOPO) and selected pharmaceutical excipients over an 8-week period under the conditions of 25 °C / 60% relative humidity and 40 °C / 75% relative humidity. The physical appearance and potency of various samples were evaluated by visual observation and high performance liquid chromatography (HPLC) analysis at T = 0, 2, 4, and 8 weeks.

[0148] HPLC system suitability and linearity were verified at each time point and included in the protocol requirements. Among the 14 compounds tested, 4 excipients (alpha starch, compressed sugar, providone, and hydrogenated vegetable oil) resulted in a decrease in the purity of 3,4,3-LI(1,2- HOPO) or an increase in the content of specific impurities. These results were taken into account for further investigation regarding the enhancement of the bioavailability of 3,4,3-LI(1,2-HOPO).

[0149] 1. Purpose of the Study The purpose of this study was to obtain data that could be used to support the results of the study. This was not conducted in accordance with the regulations of the U.S. Food and Drug Administration (FDA), "Good Laboratory Practice for Nonclinical Laboratory Studies" (GLP) as described in 21 CFR Part 58. However, this study was planned, executed, recorded, and reported in accordance with standard practices to ensure the quality and completeness of the data.

[0150] 2. Goals of the Study The goal of this study was to evaluate the interaction between 3,4,3-LI(1,2-HOPO) and selected pharmaceutical excipients over an 8-week period under the conditions of 25 °C / 60% relative humidity and 40 °C / 75% relative humidity, and included the following. a. Mannitol b. Lactose monohydrate c. Compressed sugar d. Microcrystalline cellulose e. Hypromellose f. Povidone g. Pre-gelatinized starch h. Croscarmellose sodium i. Sodium starch glycolate j. Crospovidone k. Colloidal silicon dioxide l. Magnesium stearate m. Hydrogenated vegetable oil type 1 n. Polysorbate 80 (PS), NF (Spectrum Chemicals, catalog number PO138)

[0151] 3. Experimental design The stability of 3,4,3-LI(1,2-HOPO) was tested according to the conditions listed in Table 1.1 below. All test samples were placed in 40 mL USP Type 1 clear glass vials (outer diameter 28 mm × height 95 mm, 24 mm screw cap) and stored wrapped in aluminum foil during the study.

Table 1-1

[0152] Table 1.2 shows a list of the excipients used in this study and the various drug-excipient ratios.

Table 1-2

Table 1-4

[0153] 5. Results a. System suitability The results of system suitability and linearity are summarized in Tables 1.5 - 1.8 for all time points (T = 0, 2, 4, and 8 weeks). All results of system suitability and linearity were included in the protocol requirements. The prepared calibration standard curve was found to be linear, and the correlation coefficient is included in the table along with the calibration curve.

Table 1-5

Table 1-6

Table 1-7

Table 1-8

[0154] b. Stability determination The results of the compatibility study are summarized in Tables 1.9 to 1.23. Test articles 3,4,3-LI(1,2-HOPO) were stable over 8 weeks under the conditions described (25 °C / 60% relative humidity and 40 °C / 75% relative humidity) in the control samples. Most of the excipient-API mixtures exhibited similar stability, except for the mixtures containing pre-gelatinized starch (Table 1.15) and hydrogenated vegetable oil (Table 1.21), which resulted in an apparent decrease in the HPLC purity of 3,4,3-LI(1,2-HOPO). In addition, specific increases in purity were observed for excipient-API mixtures containing pre-gelatinized starch (Table 1.15), compressed sugar (Table 1.11), povidone (Table 1.14), and hydrogenated vegetable oil (Table 1.21).

Table 1-9A

Table 1-9B

Table 1-10A

Table 1-10B

Table 1-10C

Table 1-11A

Table 1-11B

Table 1-12A

Table 1-12B

Table 1-12C

Table 1-13A

Table 1-13B

Table 1-14A

Table 1-14B

Table 1-15A

Table 1-15B

Table 1-15C

Table 1-16A

Table 1-16B

Table 1-17A

Table 1-17B

Table 1-18A

Table 1-18B

Table 1-19A

Table 1-19B

Table 1-19C

Table 1-20A

Table 1-20B

Table 1-21A

Table 1-21B

Table 1-22A

Table 1-22B

Table 1-23A

Table 1-23B

Table 1-23C

[0155] 6. Conclusion A series of commonly used pharmaceutical excipients were tested for their interaction and compatibility with 3,4,3-LI(1,2-HOPO). Among the 14 compounds tested, 4 excipients (alpha starch, compressed sugar, povidone, and hydrogenated vegetable oil) resulted in a decrease in the purity of 3,4,3-LI(1,2-HOPO) or an increase in the content of specific impurities. These 4 excipients should be avoided in future formulations of 3,4,3-LI(1,2-HOPO). (Example 2) Feasibility of Developing an Oral Formulation of 3,4,3-LI(1,2-HOPO)

[0156] Overview The feasibility of developing an oral formulation of 3,4,3-LI(1,2-HOPO) was evaluated. Four oral dosage forms were investigated: (i) powder in a bottle, (ii) dispersible / soluble granules, (iii) chewable tablets, and (iv) conventional immediate-release tablets. Based on the studies conducted, nine formulation prototypes were identified that exhibited immediate drug release behavior and the required physical properties and were selected for API verification, gastric dissolution, and testing of related substances by a specified liquid chromatography method. Of these selected compositions, two were powder formulations in a bottle, two were granule formulations, three were chewable tablet formulations, and two were conventional tablet formulations. The composition of each of these prototype formulations is summarized and presented in Table 2.1. All assays confirmed that these prototypes were suitable for further development. The stability of these formulations was evaluated prior to the first human clinical trial of 3,4,3-LI(1,2-HOPO). These stability studies also included capsules containing the powder formulation A2 in a bottle, which may be the optimal dosage form for adjusting the dosage level in a clinical setting.

Table 2-1

[0157] 1. Objectives of the study The objective of this study was to obtain data that could be used to support the results of the study. This was not conducted in accordance with the regulations of the U.S. Food and Drug Administration (FDA), "Good Laboratory Practice for Nonclinical Laboratory Studies" (GLP) as described in 21 CFR Part 58. However, this study was planned, conducted, recorded, and reported in accordance with standard practices to ensure the quality and completeness of the data.

[0158] 2. Goals of the study The objective of this study was to develop a prototype oral formulation of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO). The clinical dose of 3,4,3-LI(1,2-HOPO) is predicted to be in the range of 1 to 2 grams per unit. To maintain the flexibility to dose at lower and higher dose strengths in clinical evaluation, several oral formulations were included in the development study, including the following. - Powder in bottle (PIB) - Orally dispersible / soluble granules - Chewable tablets - Conventional oral tablets

[0159] 3. Design of experiments Suitable excipients were selected based on the results of drug-excipient compatibility studies (compatibility studies of 3,4,3-LI(1,2-HOPO)-excipients, Example 1) and evaluated for the feasibility of developing the selected formulations. All test formulations contained sodium oleate as a permeation enhancer based on the pharmacokinetic results constructed in parallel. In addition to 3,4,3-LI(1,2-HOPO), diluents, and permeation enhancers, other formulation components were also explored for each prototype formulation. The matrix of a typical formulation is shown in Table 2.2.

Table 2-2

[0160] Suitable formulation techniques, such as direct tableting, dry compaction, and / or the process of wet granulation, were evaluated. The selected prototype compositions were tested for various physicochemical properties as shown in Table 2.3.

Table 2-3

Table 2-4A

Table 2-4B

Table 2-4C

Table 2-4D

Table 2-4E

[0161] Analytical method I for the assay of related substances (Table 2.5): This method has already been established and validated (see 3,4,3-LI(1,2-HOPO)-excipient compatibility study, Example 1). Therefore, the suitability was not re-evaluated as part of this study. Equipment: Waters Alliance 2695 liquid chromatography system Detector: 2487 Waters Dual Wavelength detector Column: Agilent, Eclipse XDB-C18, 4.6×150mm, 5μm. Mobile phase A: 0.05% formic acid in 95% H2O:5% ACN Mobile phase B: 0.05% formic acid in acetonitrile (ACN) Column temperature: 25°C Flow rate: 1.0 mL / min Injection volume: 20 μL Detection: 250 nm Elution time: 50 minutes Diluent: 9:1 H2O:ACN

Table 2-5

[0162] Analytical method I for the assay of related substances (Table 2.6): This method had been previously established and validated (see 3,4,3-LI(1,2-HOPO)-excipient compatibility study, Example 1). The suitability was therefore not re-evaluated as part of this study. Instrument: Waters Alliance 2695 Liquid Chromatography System Detector: 2487 Waters Dual Wavelength Detector Column: Agilent, Eclipse XDB-C18, 4.6×150mm, 5μm. Mobile Phase A: 0.05% formic acid in 95% H2O:5% ACN Mobile Phase B: 0.05% formic acid in acetonitrile (ACN) Column Temperature: 25°C Flow Rate: 1.0 mL / min Injection Volume: 20 μL Detection: 250 nm Retention Time: 50 min Diluent: 9:1 H2O:ACN

Table 2-6

Table 2-7

Table 2-8

[0163] 5. Results a. Dosage form of powder in bottle (PIB) Powder in bottle (PIB) is one of the simplest dosage forms used in initial clinical development due to its ease of use. Compared with capsules, PIB has a large dosage and filling weight that can be held. Table 2.13 shows the composition of the evaluated PIB. The reasons for evaluating each composition are also described. The goal was to identify a suitable composition that could form a uniform dispersion when diluted with water and exhibit immediate drug release characteristics. Table 2.14 describes the corresponding characteristics of the evaluated formulations.

Table 2-13

Table 2-14

[0164] API 3,4,3-LI(1,2-HOPO), and all other compositions except A9 - A11 formed viscous masses when reconstituted in water. This behavior was significantly reduced after incorporating Avicel RC-591 (a co-processed excipient of microcrystalline cellulose and carboxymethylcellulose). Composition A11 prepared using a 1:1 drug:excipient ratio formed a uniform dispersion and was therefore evaluated using an API content verification assay and a dissolution assay in simulated gastric fluid without enzymes. Both assays were performed using Composition A2 (a blend of API + the permeation enhancer sodium oleate) for comparison, as further described in this report. b. Orally dispersible / soluble granules

[0165] The oral dispersible / soluble granule is similar to the commercially available "Sprinkles", where the granules of 3,4,3-LI(1,2-HOPO) are directly carried into the mouth from individually packaged pouches / sachets and can be swallowed with or without water. Table 2.15 shows the various compositions of the evaluated oral dispersible / soluble granules. The reasons for evaluating each composition will also be described.

Table 2-15A

Table 2-15B

[0166] The goal was to identify a suitable composition that could provide a smooth feel in the mouth and exhibit immediate drug release characteristics. Table 2.16 describes the corresponding properties of the evaluated formulations.

Table 2-16

[0167] Among the various compositions evaluated, G11 formulated using a co-processed product of microcrystalline cellulose (Avicel CE-15) showed a smooth feel when wetted with water. Composition G12 formulated using lactose monohydrate (Pharmatose 300M) also showed a smooth feel after several minutes of wetting. Based on these observations, compositions G11 and G12 were further tested for API verification assays and dissolution assays in simulated gastric fluid without enzymes. The results are further presented in this report. c. Chewable tablets

[0168] Chewable tablets are formulated for oral use. They are usually uncoated and formulated to provide release and absorption of the active ingredient in the mouth / buccal side or from the stomach. Table 2.17 shows the compositions of the various chewable tablets evaluated. The goal was to identify a suitable composition that could be formulated by a direct tableting process and also exhibit immediate drug release characteristics. Table 2.18 describes the corresponding properties of the formulations evaluated.

[0169] Compositions C13 and C21 showed satisfactory physical properties (absence of segregation, friability, and disintegration). These compositions, along with composition C11, were further tested for API verification assay and dissolution assay in simulated gastric fluid without enzymes. The results are further presented in this report.

Table 2-17A

Table 2-17B

Table 2-17C

Table 2-18A

Table 2-18B

[0170] Tablet compositions T44, T45, T50, and T51 exhibited the ideal tablet characteristics (tabletability, friability, hardness, and disintegration). Compositions T44 and T45 were prepared by the wet granulation process, and compositions 50 and 51 were prepared by direct tableting. Generally, based on the aspects of drug stability, manufacturing time, and cost, direct tableting is a preferred process. Therefore, compositions T50 and T51 were considered ideal, and their validation assays and dissolution in simulated gastric fluid without enzymes were tested. The results are further presented in this report.

Table 2-19A

Table 2-19B

Table 2-19C

Table 2-20A

Table 2-20B

Table 2-20C

Table 2-21A

Table 2-21B

Table 2-21C

Table 2-22A

Table 2-22B

Table 2-22C

[0171] e. Selected prototype: Appearance and API verification assay Based on the development studies conducted, nine formulation prototypes that exhibited immediate drug release behavior and the required physical properties were identified and selected for further testing: powder compositions A2 and A11 in bottles, granule compositions G11 and G12, chewable tablet compositions C11, C13, and C21, and immediate release tablet compositions T50 and T51. The appearance and packaging of the selected powder formulation prototypes A2 and A11 in bottles are shown in Figures 7 (left) and 7 (right), respectively. The appearance and packaging of the selected granule formulation prototypes G11 and G12 are shown in Figures 8 (left) and 8 (right), respectively. The appearance and packaging of the selected chewable tablet formulation prototypes C11, C13, and C21 are shown in Figures 9 (left), 9 (center), and 9 (right), respectively. The appearance and packaging of the selected tablet formulation prototypes T50 and T51 are shown in Figures 10 (left) and 10 (right), respectively.

[0172] These selected prototype formulations were assayed for their 3,4,3-LI(1,2-HOPO) content according to the method outlined for the content verification assay. Generally, all of the formulations tested were found to contain 90 - 110% of the labeled 3,4,3-LI(1,2-HOPO). Table 2.23 lists the values obtained for the verification assay for each of them.

Table 2-23

[0173] In the preparation of samples from chewable tablets, the matrix material of the tablets gels upon exposure to the diluent, thereby preventing complete extraction into the assay medium by either shaking (using a wrist action shaker) or sonication. For this reason, the tablets were crushed using a mortar and pestle. Crushing of the tablet matrix assisted in the complete extraction of the analyte. Extraction from intact dosage forms in other prototype formulations was complete, as observed from the values in Table 2.23. f. Selected prototype: dissolution assay

[0174] The in vitro dissolution tests of the prototype formulations were carried out as detailed in the methods section. Generally, more than 80% of the API was released within 45 minutes of the dissolution test for all formulations tested. The results of the study are listed in Tables 2.24 to 2.27.

Table 2-24

Table 2-25

Table 2-26

Table 2-27

[0175] g. Selected prototype: assay for related substances The related substances predicted as area (%) obtained from the chromatogram and the chromatographic purity of 3,4,3-LI(1,2-HOPO) in the selected prototype formulation are listed in Table 2.28. For all the compositions tested, the amount of related substances found in the prototype formulation is equivalent to those present in the drug substance used as a control.

Table 2-28A

Table 2-28B

[0176] 6. Conclusion The feasibility of developing an oral formulation of 3,4,3-LI(1,2-HOPO) was evaluated. Four oral dosage forms were investigated: (i) powder in bottle, (ii) dispersible / soluble granules, (iii) chewable tablets, and (iv) conventional immediate-release tablets. Based on the studies conducted, nine formulation prototypes showing immediate drug release behavior and the required physical properties were identified and selected for API verification, gastric dissolution, and testing of related substances by a specified liquid chromatography method. Of these selected compositions, two are powder formulations in bottle, two are granule formulations, three are chewable tablet formulations, and two are conventional tablet formulations. All assays confirmed that these prototypes are suitable for further development. The stability of these formulations will be evaluated prior to the first human clinical trial of 3,4,3-LI(1,2-HOPO). These stability studies also include capsules containing the powder composition A2 in bottle, which may be the optimal dosage form for adjusting the dosage level in a clinical setting. (Example 3) Evaluation of the Stability of Prototype Oral Formulations of the Active Pharmaceutical Ingredient 3,4,3-LI(1,2-HOPO)

[0177] Overview The objective of this study was to evaluate the stability of a prototype oral formulation of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO) over a six-month period under storage conditions of 25°C / 60% relative humidity and 40°C / 75% relative humidity. The clinical dose of 3,4,3-LI(1,2-HOPO) is predicted to be in the range of 1 to 2 grams per unit. To maintain the flexibility to dose at lower or higher dose strengths in clinical evaluations, several oral formulations were included in this study. - Reconstitution powder (500 mg) - Immediate release tablets (500 mg) - Chewable tablets (500 mg) - Capsules (500 mg) - Capsules (100 mg) - Placebo capsules (size 00) - Placebo capsules (size 4)

[0178] The following is a summary of the observations and trends observed in the stability study of the prototype formulations of 3,4,3-LI(1,2-HOPO). · When stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity for six months, there were no changes in the physical appearance of the prototype formulations and placebo. · In the reconstitution powder prototype formulation stored at 40°C / 75% relative humidity, the water content increased slightly. In all other formulations, including the placebo, the water content was equivalent to the value observed in the T0 sample. · The hardness of the chewable tablets and immediate release tablets decreased slightly during storage compared to the values at T0. · When stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity, the occurrence of rancid odor was observed in all prototype formulations except the reconstitution powder. The occurrence of rancid odor was also observed in the placebo of the 3,4,3-LI(1,2-HOPO) capsules when stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity. · In the stability study samples, no significant changes were observed in the dissolution profiles of the prototype formulations. The dissolution of 3,4,3-LI(1,2-HOPO) from the chewable tablets and immediate-release tablets was found to be slightly faster in the stability study samples than that observed in the T0 samples. All the samples tested released more than 85% of the labeled 3,4,3-LI(1,2-HOPO) within 45 minutes. · In the assay of the labeled % of 3,4,3-LI(1,2-HOPO) in the prototype dosage forms, it was found to be 90 - 110% in all the stability study samples analyzed, as well as in the T0 samples. · The chromatographic purity measured for the prototype formulations in the stability study changed slightly in the samples stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity compared to that observed at T0.

[0179] 1. Purpose of the study The purpose of this study was to obtain data that could be used to support the results of the study. This was not conducted in accordance with the regulations of the U.S. Food and Drug Administration (FDA), "Good Laboratory Practice for Nonclinical Laboratory Studies" (GLP) as described in 21 CFR Part 58. However, this study was planned, conducted, recorded, and reported in accordance with standard practices to ensure the quality and completeness of the data.

[0180] 2. Objectives of the study The objective of this study was to evaluate the stability of a prototype oral formulation of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO) over a 6-month period under storage conditions of 25°C / 60% relative humidity and 40°C / 75% relative humidity. The clinical dose of 3,4,3-LI(1,2-HOPO) is predicted to be in the range of 1 to 2 grams per unit. To maintain the flexibility to dose at lower or higher dose strengths in clinical evaluations, several oral formulations were included in this study. - Powder for reconstitution (500 mg) - Immediate-release tablets (500 mg) - Chewable tablets (500 mg) - Capsules (500 mg) - Capsules (100 mg) - Placebo capsules (size 00) - Placebo capsules (size 4)

[0181] 3. Experimental design Prototype formulations were selected based on the results of the aforementioned formulation development study (3,4,3-LI(1,2-HOPO)-formulation development, Example 2) and are presented in Tables 3.1 and 3.2 below.

[0182] All prototype formulations and placebo capsules were sampled at 1 month, 3 months, 6 months, and T0 (initial) over a 6-month period under storage conditions of 25 ± 2°C / 60 ± 5% relative humidity or 40 ± 2°C / 75 ± 5% relative humidity and staged. Table 3.3 shows the packaging configurations of all prototype formulations.

[0183] Various tests were applied to each sample at each sampling time point as detailed in Table 3.4.

Table 3-1A

Table 3-1B

Table 3-2

Table 3-3

Table 3-4

Table 3-5

Table 3-6

Table 3-7

[0184] 5. Results The results of all tests conducted on the prototype formulation are summarized in Tables 3.8 to 3.12, and each table shows the results of a specific assay.

Table 3-8A

Table 3-8B

Table 3-8C

Table 3-8D

Table 3-9A

Table 3-9B

Table 3-9C

Table 3-9D

Table 3-9E

Table 3-10A

Table 3-10B

Table 3-11A

Table 3-11B

Table 3-12A

Table 3-12B

[0185] 6. Conclusion The following is an overview of the observations and trends observed in the stability study of the prototype formulation of 3,4,3-LI(1,2-HOPO). · When stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity for 6 months, there were no changes in the physical appearance of the prototype formulation and the placebo. · In the reconstituted powder prototype formulation stored at 40°C / 75% relative humidity, the water content increased slightly. In all other formulations including the placebo, the water content was equivalent to the value observed in the T0 sample. · The hardness of the chewable tablets and immediate release tablets decreased slightly during storage compared to the values at T0. · When stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity, the generation of rancid odor was observed in all prototype formulations except the reconstituted powder. The generation of rancid odor was also observed in the placebo of the 3,4,3-LI(1,2-HOPO) capsules when stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity. · In the stability study samples, no significant changes were observed in the dissolution profiles of the prototype formulations. The dissolution of 3,4,3-LI(1,2-HOPO) from the chewable tablets and immediate release tablets appears to be slightly faster in the stability study samples than that observed in the T0 samples. All the samples tested released more than 85% of the labeled 3,4,3-LI(1,2-HOPO) within 45 minutes. · In the assay of the labeled % of 3,4,3-LI(1,2-HOPO) in the prototype dosage form, it was found to be 90 - 110% in all the stability study samples analyzed, as well as in the T0 samples. · The chromatographic purity measured for the prototype formulation in the stability study changed slightly compared to that observed at T0 for the samples stored at 25°C / 60% relative humidity and 40°C / 75% relative humidity. (Example 4) From the bodies of female and male Swiss Webster mice, 238 Effectiveness of repeated 3,4,3-LI(1,2-HOPO) treatment for removing an intravenous dose of Pu

[0186] Summary The goal of this study was to 238 characterize the effectiveness of 3,4,3-LI(1,2-HOPO) in enhancing the excretion from internal plutonium burden in female and male Swiss Webster mice that received a soluble citrate complex of Pu and underwent multiple treatments starting 24 hours after exposure. The tissue content of Pu and the excretion via urine and feces in the treated animals were compared to those in the untreated animals to evaluate the effectiveness. 238

[0187] The medical treatment 3,4,3-LI(1,2-HOPO) was administered either parenterally (intraperitoneally) or orally (orally) via continuous injection or tube feeding (once daily for 6 times or twice daily for 12 times) starting 24 hours after contamination at the following four selected doses: 30 μmol / kg intraperitoneally, 150 μmol / kg orally, 300 μmol / kg orally, and 600 μmol / kg orally (corresponding to approximate human dose ranges of 2.5, 12.5, 25, and 50 μmol / kg, respectively, using an acceptable conversion system to human equivalent dose, HED, based on body surface area and mouse doses). The oral doses also included a permeation enhancer selected during formulation development studies.

[0188] Repeated parenteral and oral treatments with 3,4,3-LI(1,2-HOPO) resulted in enhanced excretion rates and reduced total body burdens and different tissue contents even when the first treatment dosing was delayed until 24 hours after contamination. In the first cohort euthanized at day 7, obtained with a twice-daily dosing scheme 238 Pu excretion was not as good as the corresponding once-daily dosing scheme with an equivalent total daily amount of API when compared to the saline control (i.e., once-daily dosing at 300 and 600 μmol / kg was better than twice-daily dosing at 150 and 300 μmol / kg). By extending the dosing regimen from single-dose to up to 6 times once-daily dosing, a more sustained excretion rate was possible in the group treated with 3,4,3-LI(1,2-HOPO) compared to the saline-administered control. At 11 days after contamination, the maximum in-vivo removal effect was observed after 6 times once-daily parenteral dosing of 3,4,3-LI(1,2-HOPO). After multiple oral treatments 238 The enhancement of Pu excretion remained dose-dependent since the reduction of in-vivo and tissue contents was slightly greater after dosing at 600 μmol / kg 6 times once-daily than after the corresponding dosing regimen at 300 μmol / kg. In any case, oral treatment at 300 μmol / kg was significant compared to the saline-treated control 238It resulted in a reduction in the whole body and tissue content of Pu, and the in vitro removal effect was equivalent to that of parenteral treatment with DTPA. Finally, differences in the excretion pathways were observed, 238 Pu excretion occurred mainly through feces in mice treated with 3,4,3-LI(1,2-HOPO) and through urine in mice treated with DTPA. In females, the 238 Ratio of fecal Pu was lower compared to males.

[0189] Based on the results of this study, the effective dose level of oral treatment administration was confirmed. When formulated with sodium oleate and orally administered once a day for 6 consecutive days, 3,4,3-LI(1,2-HOPO) at a dose level of 300 - 600 μmol / kg resulted in a significant 238 in vitro removal effect of soluble Pu in mice.

[0190] Objectives of the study The objective of this study was to 238 characterize the effectiveness of 3,4,3-LI(1,2-HOPO) in enhancing the excretion from internal plutonium burden in female and male Swiss Webster mice that received a soluble citrate complex of Pu and underwent multiple treatments starting 24 hours after exposure. The effectiveness was evaluated by comparing the 238 tissue content of Pu and its excretion by urine and feces in treated animals with those in untreated animals.

[0191] In this regimen optimization study, the medical agent 3,4,3-LI(1,2-HOPO) was administered either intraperitoneally (ip) or orally (po) via continuous injection or intragastric administration (once daily for 6 times or twice daily for 12 times) starting 24 hours after contamination at the following four selected doses: 30 μmol / kg intraperitoneally, 150 μmol / kg orally, 300 μmol / kg orally, and 600 μmol / kg orally (corresponding to approximate human dose ranges of 2.5, 12.5, 25, and 50 μmol / kg, respectively, using an acceptable conversion system from mouse doses to human equivalent doses [HED] based on body surface area). Currently, the “clinical oral dose” of this product obtained from preliminary studies is in the range of 10 - 150 μmol / kg when administered orally once at 24 hours after isotope internalization. The selected doses correspond to the lowest doses that result in approximately maximal in vitro removal effects when administered once at 24 hours after exposure. These doses have not caused any obvious toxicity in previous experiments. Experimental Design Test Isotope: 238 Pu Test Dose: 25 nCi (i.e., approximately 0.8 μCi / kg) Route of Contamination: Intravenous (iv), tail vein Route of Treatment: Intraperitoneal injection (ip) or oral (po) Frequency of Treatment: Starting 24 hours after contamination, multiple (once daily or twice daily for 6 days) dosing. Calculation of treatment dose: The dose of the in vitro removal agent (mg / kg or μmol / kg) was based on the body weight of the individual measured after contamination. Duration of Study: 11 days, survival status

Table 4-1

[0192] f. Control of bias During the evaluation of the animals' responses and the execution of the analysis, the technical staff was aware of the treatment history of each animal and sample. However, based on the relatively objective endpoints to be tested, bias was not expected to affect the results of the study.

[0193] 6. Results The in-life portion of the study was successfully achieved. The mean radiochemical recovery rates in the female arm were 95.6% and 88.5% for the necropsy cohorts on days 7 and 11, respectively. The mean radiochemical recovery rates in the male arm were 85.7% and 86.5% for the necropsy cohorts on days 7 and 11, respectively.

[0194] No adverse events were observed in animal groups treated with parenteral or oral 3,4,3-LI(1,2-HOPO) at all dose levels. All female and male animals in all dose groups appeared healthy and survived until their respective scheduled necropsies, except for one female mouse that was found dead in her cage 4 hours after the first treatment (parenteral 3,4,3-LI(1,2-HOPO) treated group at 30 μmol / kg once daily, necropsy scheduled for day 11). During necropsy, a large blood clot was found in the left abdominal wall, and internal bleeding due to a failed dose administration was the presumed cause of death. This mouse was not included in the mean dose calculations for the corresponding group. Several anomalies were noted during sample collection and processing, but did not significantly affect the outcome of the study: urine samples were spilled for two female mice on a single day of collection (on day 8 for one female mouse in the 300 μmol / kg oral treatment group scheduled for necropsy on day 11, and on day 7 for one female mouse in the 600 μmol / kg treatment group scheduled for necropsy on day 11).

[0195] A. Female fecal data analysis For all treatment groups with necropsy scheduled 7 days after contamination, Figure 11A shows daily fecal 238 Figure 11B shows the daily urinary excretion of Pu. 238 For all treatment groups scheduled for necropsy 11 days after contamination, Figure 12A shows the daily fecal Pu excretion. 238 Figure 12B shows the excretion of Pu by daily urine. 238 Pu excretion by cumulative urine and feces at different scheduled necropsy time points (7 and 11 days after contamination, respectively). 238 Pu excretion is shown graphically in Figures 11C and 12C and numerically in Table 4.2A for all experimental groups. All results are based on total recovery. 238 Expressed as a percentage of Pu dose.

[0196] Statistical analysis was not performed on a daily basis but was performed on cumulative fecal, urinary, and combined excretion data at the two scheduled necropsy time points. Oral treatment with 3,4,3-LI(1,2-HOPO) at 150 and 300 μmol / kg twice daily, or 300 and 600 μmol / kg once daily, resulted in a significant 238 enhancement of Pu excretion at the time point 7 days after contamination, and the groups treated at the higher 3,4,3-LI(1,2-HOPO) dose levels of 300 μmol / kg twice daily and 300 and 600 μmol / kg once daily also showed a significant enhancement of fecal excretion at the 7-day time point. At the necropsy time point 11 days later, all treatment regimens showed a significant enhancement of combined excretion. However, only the i.p. treatment at 30 μmol / kg and the oral treatment at 600 μmol / kg resulted in a significant enhancement of excretion through both urine and feces. The once-daily treatment was seen to enhance urinary excretion compared to a single treatment with 3,4,3-LI(1,2-HOPO) (see previous reports).

Table 4-2A

Table 4-2B

[0197] Figures 13A and 13B show the cumulative urinary, fecal, and combined Pu excretion of all treatment groups compared to the control at days 7 and 11, respectively, and Table 4.2B summarizes the percentage enhancement of excretion through urine, feces, and combined compared to the untreated control group. The total excretion rate was better after oral treatment with 600 μmol / kg of 3,4,3-LI(1,2-HOPO) six times once daily than after i.p. DTPA, and at the time point 11 days after contamination, it resulted in a maximum 143% increase in total excretion compared to the control group. Finally, the twice-daily dosing at 150 and 300 μmol / kg was 238 less effective than the corresponding once-daily dose levels of 300 and 600 μmol / kg. 238Dose splitting appeared to reduce efficacy as it resulted in lower enhancement of Pu excretion.

[0198] b. Female tissue data analysis At autopsy time points of day 7 (FIG. 14A) and day 11 (FIG. 14B), 238 Whole body, skeletal, and liver contents of Pu are shown graphically and numerically in Table 4.3A for all experimental groups. All results are based on total recovery. 238 All tissues analyzed showed significantly greater increases in schizophrenia compared to the corresponding saline control groups for groups treated with DTPA or 3,4,3-LI(1,2-HOPO) at all dosing regimens. 238 All groups treated with 3,4,3-LI(1,2-HOPO) showed a significant reduction in tissue Pu content compared to the saline control group. 238 Groups treated with 300 and 600 μmol / kg 3,4,3-LI(1,2-HOPO) once daily showed significant reductions in Pu total body content, and those treated with 300 and 600 μmol / kg 3,4,3-LI(1,2-HOPO) once daily showed significant reductions in Pu content in the liver, kidney, gastrointestinal tract, soft tissue, and skeleton. Finally, oral treatment with 300 μmol / kg 3,4,3-LI(1,2-HOPO) once daily produced an ex vivo removal effect equivalent to that of parenteral treatment with DTPA.

[0199] Table 4.3B shows the percentage of tissue content reduction (for significant reduction) compared to the corresponding untreated control group. All treatment groups showed significant reduction, with up to a 45% reduction in body burden compared to the control group after once-daily oral treatment with 600 μmol / kg 3,4,3-LI(1,2-HOPO) at 11 days post-contamination. Data relating to percent recovered dose from the female arm are shown in Table 4.6. [Table 4-3A-1] [Table 4-3A-2] [Table 4-3B]

[0200] c. Male fecal data analysis For all treatment groups with necropsy scheduled 7 days after contamination, Figure 15A shows daily fecal 238 Figure 15B shows the excretion of Pu by daily urine. 238 For all treatment groups scheduled for necropsy 11 days after contamination, Figure 16A shows the daily fecal Pu excretion. 238 Figure 16B shows the excretion of Pu by daily urine. 238 Pu excretion by cumulative urine and feces at different scheduled necropsy time points (7 and 11 days after contamination, respectively). 238 Pu excretion is shown graphically in Figures 15C and 16C and numerically in Table 4.4A for all experimental groups. All results are based on total recovery. 238 Oral treatment with 150 and 300 μmol / kg twice daily or 300 and 600 μmol / kg once daily of 3,4,3-LI(1,2-HOPO) significantly reduced the risk of infection at 7 days after contamination. 238 All treatment regimens showed significant combined excretion and enhanced fecal excretion at necropsy 11 days later. However, oral treatment with 3,4,3-LI (1,2-HOPO) did not result in significant enhancement of urinary excretion. [Table 4-4A] [Table 4-4B]

[0201] Figures 17A and 17B show cumulative urinary, fecal, and combined morbidity for all treatment groups compared to controls on days 7 and 11, respectively. 238Figure 4.4B shows the excretion of Pu and summarizes the rates of enhanced excretion in urine, feces, and the combination relative to the untreated control group. The total excretion rate was better after oral treatment with 600 μmol / kg of 3,4,3-LI(1,2-HOPO) once a day for 6 days than after parenteral DTPA, and at the 11-day time point of contamination, compared to the control group, it resulted in a maximum 156% increase in total excretion. Twice-daily dosing at 150 μmol / kg resulted in lower enhancement of Pu excretion than the corresponding once-daily 300 μmol / kg dose level, and thus dose splitting was seen to reduce efficacy at lower dose levels. However, twice-daily dosing at 300 μmol / kg or once-daily dosing at 300 μmol / kg resulted in equivalent 238 enhancement of Pu excretion. One difference that was seen to occur upon splitting was 238 a change in the urine:feces ratio of Pu, with excretion by urine being higher after split doses (twice-daily regimen) than after the corresponding once-daily dosing regimen, which may indicate saturation of the liver's clearance capacity. d. Analysis of male tissue data 238 The whole-body, skeletal, and liver contents of Pu at necropsy time points on day 7 (Figure 18A) and day 11 (Figure 18B) are graphed for all experimental groups and presented numerically in Table 4.5A. All results are expressed as a percentage of the total recovered

[0202] Pu dose. All tissues analyzed showed a substantial reduction in Pu tissue content for groups treated with DTPA or 3,4,3-LI(1,2-HOPO) compared to the corresponding saline control group at all dosing regimens. All groups treated with 3,4,3-LI(1,2-HOPO) showed 238 a substantial reduction in Pu tissue content compared to the saline control group. 238 238 238 ​​The whole-body content of Pu showed a significant reduction, and the groups treated with 3,4,3-LI(1,2-HOPO) at 300 and 600 μmol / kg once a day showed significant reductions in liver, kidney, gastrointestinal tract, soft tissue, and skeletal content. Finally, oral treatment with 3,4,3-LI(1,2-HOPO) at 300 and 600 μmol / kg once a day resulted in an in vitro removal effect equivalent to that of parenteral treatment with DTPA.

[0203] Table 4.5B shows the percentage reduction in tissue content (for significant reductions) compared to the corresponding untreated control group. All treatment groups showed significant reductions, and at the 11-day time point of contamination, after once-daily oral treatment with 600 μmol / kg of 3,4,3-LI(1,2-HOPO), there was a maximum 48% reduction in body burden compared to the control group. Data related to the percent of the recovered dose obtained from the male arm are shown in Table 4.7.

[0204] Generally, the reduction in tissue burden follows a similar pattern in male and female animals.

Table 4-5A-1

Table 4-5A-2

Table 4-5B

[0205] 7. Conclusions Repeated parenteral and oral treatment with 3,4,3-LI(1,2-HOPO) resulted in enhanced excretion rates and reduced total body burden and different tissue contents even when the first treatment dose was delayed until 24 hours after contamination. In the first cohort euthanized at the 7-day time point, obtained with a twice-daily dosing scheme 238Pu excretion was not as good as the corresponding once-daily dosing scheme using the same total daily amount of API compared to the saline control (i.e., once-daily dosing at 300 and 600 μmol / kg was better than twice-daily dosing at 150 and 300 μmol / kg). By extending the dosing regimen from single-dose to up to six once-daily doses, a more sustained excretion rate was possible in the group treated with 3,4,3-LI(1,2-HOPO) compared to the saline-administered control. The maximum in vivo removal effect was observed at 11 days post-contamination after six i.p. once-daily doses of 3,4,3-LI(1,2-HOPO). Enhancement of 238 Pu excretion remained dose-dependent since the reduction in body and tissue burdens was slightly greater after six once-daily doses of 600 μmol / kg compared to the corresponding 300 μmol / kg dosing regimen. Nevertheless, oral treatment at 300 μmol / kg resulted in a significant 238 reduction in the total body and tissue burdens of Pu and an in vivo removal effect comparable to that of i.p. treatment with DTPA. Finally, differences in the excretion pathways were noted, 238 and Pu excretion occurred mainly via feces in mice treated with 3,4,3-LI(1,2-HOPO) and via urine in mice treated with DTPA, with a lower fecal-to-urine ratio for Pu in females compared to males. 238

[0206] The results of this study identified an effective dose level for oral treatment administration. When formulated with sodium oleate and orally administered once daily for six consecutive days, 3,4,3-LI(1,2-HOPO) at dose levels of 300 - 600 μmol / kg produced a significant 238 in vivo removal effect of soluble Pu in mice. a. Percent recovered dose in the female arm

Table 4-6-1

Table 4-6-2

Table 4-6-3

Table 4-6-4

Table 4-6-5

Table 4-6-6

Table 4-6-7

Table 4-6-8

Table 4-6-9

Table 4-6-10

Table 4-6-11

Table 4-6-12

Table 4-6-13

Table 4-6-14

Table 4-6-15

Table 4-6-16

Table 4-6-17

Table 4-6-18

Table 4-6-19

Table 4-6-20

Table 4-7-1

Table 4-7-2

Table 4-7-3

Table 4-7-4

Table 4-7-5

Table 4-7-6

Table 4-7-7

Table 4-7-8

Table 4-7-9

Table 4-7-10

Table 4-7-11

Table 4-7-12

Table 4-7-13

Table 4-7-14

Table 4-7-15

[0207] In Examples 5 to 9, the treatment dose levels of each in vivo study are expressed in μmol / kg and / or mg / kg. Based on the molecular weight of API 3,4,3-LI(1,2-HOPO) of 750.71 g / mol, the mg / kg dose level is divided by 0.7507 to obtain the dose level in μmol / kg units, while the dose level in μmol / kg units is multiplied by 0.7507 to obtain the dose level in mg / kg units. The dose levels commonly used in non-clinical studies are presented in both μmol / kg and mg / kg in Table 0.1 for reference purposes.

Table A

[0208] The formulated material was used in single-dose in vivo ADME characterization studies in Sprague-Dawley rats and Swiss Webster mice using 14C-labeled 3,4,3-LI(1,2-HOPO).

[0209] The pharmacokinetic parameters and disposition / biodistribution of 3,4,3-LI(1,2-HOPO) were characterized in vivo (using two 14C labels on the spermine backbone of the parent product) in two non-GLP studies using Swiss Webster mice and Sprague-Dawley rats. The study designs for these two studies are shown in Table 5.1 and Table 5.2, respectively. Groups of six mice (three males and three females) were each dosed once with [14C]-3,4,3-LI(1,2-HOPO) via the intravenous, intraperitoneal, or oral route. A group of an additional six mice (three males and three females) was dosed with 14C-labeled API by oral gavage (po) together with a permeation enhancer (10% sodium oleate). Similarly, groups of six rats (three males and three females) were dosed with a single intravenous dose of 14C-labeled API or a single oral dose of 14C-labeled API together with 10% sodium oleate. In each of these studies, samples were collected at scheduled time points up to 24 hours post-dose at most and analyzed for 14C content using liquid scintillation counting. Blood, liver, kidney, feces, and urine were collected from the mice and analyzed. Blood, brain, liver, kidney, lung, spleen, skeletal muscle tissue, gastrointestinal (GI) tract samples, carcass, feces, and urine were collected from the rats and analyzed.

Table 5-1

Table 5-2

[0210] The pharmacokinetic parameters determined from these studies are presented in Table 5.3. Blood was collected at 6 - 8 time points from 5 minutes to 24 hours after dosing. The plasma concentration - time profiles showed similar log - linear decay after intravenous administration in mice and rats, and the radiolabeled compound rapidly distributed throughout the extracellular fluid space, with peak concentrations and total plasma exposures being higher in rats (in males and females, C0 = 463 and 422 Pg - eq / ml, AUC = 354, and 211 hour Pg - eq / ml respectively) than in mice (in females and males, C0 = 342 and 76 Pg - eq / mL, AUC = 66.2, and 41.7 hour Pg - eq / ml respectively). Radioactivity was cleared from plasma with t1 / 2 values of approximately 1.6 hours and 8 hours for mice and rats respectively after intravenous administration. Intraperitoneal administration of [14C] - 3,4,3 - LI(1,2 - HOPO) in mice resulted in plasma radioactivity levels that were lower than the intravenous route but significantly higher than the oral route. The oral bioavailability (F) of 3,4,3 - LI(1,2 - HOPO) was limited as indicated by its low plasma exposure. The bioavailability of the radioactive compound was slightly higher in females compared to males as shown in Table 5.3 (2.6% vs 1.2% in male and female mice respectively, 1.1% vs 0.4% in male and female rats respectively). Formulating 3,4,3 - LI(1,2 - HOPO) with sodium oleate resulted in a moderate improvement in systemic exposure. Cmax was improved approximately 3 - fold, from 0.32 to 0.93 Pg - eq / ml in male mice and from 0.55 to 1.4 Pg - eq / ml in female mice. In addition, the AUC increased from 8.3 ± 6.2 to 17.4 ± 6.7 min Pg - eq / ml in males and from 23.0 ± 15.4 to 35.1 ± 18.9 min Pg - eq / ml in females when calculated over 2 hours after treatment, which was interpreted as an approximately 2 - fold improvement in oral bioavailability in mice.

Table 5 - 3

[0211] Analysis of the radioactivity levels in tissues 14 showed that radioactivity from [11C]-3,4,3-LI(1,2-HOPO) was rapidly distributed to the liver and kidneys after intravenous injection. Generally, the highest levels of radioactivity in the kidneys and liver were detected early, 1 hour after dosing in mice and 2 hours after dosing in rats (Figures 19A - 19F and Figures 20A - 20D). A similar trend was observed in intraperitoneally injected mice, where the highest levels of radioactivity in the kidneys and liver were detected 1 hour after dosing. No significant difference in radioactivity uptake into the liver and kidneys was observed between the intravenous and intraperitoneal routes of administration in mice, and both routes 14It has been shown to be effective in the distribution of C]-3,4,3-LI(1,2-HOPO). In the intravenous treatment group of rats, the general order of tissue concentrations at the 2-hour time point was kidney > liver > lung > brain ≒ spleen ≒ muscle. The tissue radioactivity levels of the liver and kidney were similar and remained high at all time points after intravenous administration, while the levels in other tissues decreased more rapidly after 2 hours. As a result, after intravenous administration, radioactivity tended to concentrate in the excretory organs (kidney and liver), and therefore, the tissue-to-plasma ratio increased with time in these tissues. In contrast, after oral dosing, the highest concentrations of radioactivity were observed in feces and the gastrointestinal tract, and very low levels of radioactivity were observed in urine, plasma, and other tissues (Figs. 19A - 19F and Figs. 20A - 20D).

[0212] The major excretion routes of radioactivity from [[14C]]-3,4,3-LI(1,2-HOPO) are both feces and urine, due to high levels of radioactivity in both feces and urine, as well as significant radioactivity in the excretory tissues, the kidneys, liver, and gastrointestinal tract, after intravenous dosing. Excretion by feces up to 24 hours after dosing accounted for approximately 62% and 16% of the intravenously administered dose in mice and rats, respectively. Excretion by urine was approximately 12 - 23% of the intravenously administered dose in these rodents, and excretion of [[14C]] by the kidneys was initiated early, about 5 minutes after intravenous injection, in mice. In contrast, after oral dosing, excretion was mainly via the fecal route and accounted for approximately 89% and 41% of the orally administered dose in mice and rats, respectively, up to 24 hours after dosing. Excretion by urine, less than 1% of the orally administered dose, and low levels of radioactivity were detected in tissues excluding the systemic circulation and gastrointestinal tract. In mice, after all three routes of administration (i.e., intravenous, intraperitoneal, and oral), the highest [[14C]] accumulation was seen in feces, and the biliary route was confirmed to be the main mode of excretion, at least for the intravenous and intraperitoneal routes of administration. Based on the 15.5-hour colonic transit time in rats, the radioactivity found in feces in the first 24 hours after oral dosing is most likely the unabsorbed compound. Following metabolism in the liver, excretion in bile is possible for orally bioavailable 3,4,3-LI(1,2-HOPO), but due to very low levels of radioactivity in the blood and tissues in animals in the oral group, the major route of excretion of orally dosed 3,4,3-LI(1,2-HOPO) is through feces, suggesting that it is composed of the unabsorbed parent compound and metabolites resulting from first-pass effects in the liver or biotransformation in the gastrointestinal tract. Profiling of the metabolites of [[14C]]-3,4,3-LI(1,2-HOPO) was performed using an HPLC method on selected urine, feces, kidney, liver, and lung samples from Sprague-Dawley rats. Only samples with the highest total radioactivity levels were selected for analysis.A total of 11 peaks were detected, 6 of which were also seen in the spiked blank matrix control and were thus due to uncharacterized interactions between the test article and matrix components. These 6 peaks were considered to be not metabolites but another form of the parent compound (e.g., a complex with metal ions of the test article). The other 5 radionuclides (peaks P1 - P4 and P10) were considered to potentially be metabolites. The feces-specific metabolite peaks P2, P3, and P10 together represented 10.5 - 11.4% and 0.5 - 4.2% of the administered dose, respectively, after oral or intravenous administration in rats. Peak P10 was the most abundant peak in all samples analyzed. This peak represented up to 10% of the administered dose in feces after oral administration, while the other two feces-specific peaks represented less than 1% of the administered dose after oral administration. Peak P10 was also dominant and was sometimes the only peak in fecal samples after intravenous administration. Since P10 was not detected in liver samples, it could be of biliary origin or highly likely to be a product of conversion in the intestinal tract either through spontaneous degradation processes or mediated by the intestinal flora. P1 was the only metabolite peak identified in urine, representing less than 0.4% of the intravenously administered dose and was not present in the single urine sample analyzed after oral administration. In conclusion, evaluation of the metabolite profile showed that the putative major metabolite of [14C]-3,4,3-LI(1,2-HOPO) that accounted for approximately 10% of the administered oral dose was formed (P10). Therefore, the observed low bioavailability of [14C]-3,4,3-LI(1,2-HOPO) was highly likely due to both a likely in vivo conversion process in the gastrointestinal tract and relatively low absorption after oral administration. (Example 6) GLP Single-Dose Oral Safety by Pharmacokinetic Evaluation Study in Beagle Dogs

[0213] Pharmacokinetic parameters in beagle dogs after single oral administration of the formulated capsules were determined in a GLP study. Using a validated bioanalytical method, plasma concentrations were determined in three dogs / sex after administration of capsules of the clinical formulation of 3,4,3-LI(1,2-HOPO) at 37.5, 75, and 150 mg / kg (50, 100, and 200 μmol / kg). At all three dose levels, plasma concentrations reached a peak 0.6 - 1.1 hours after dosing (Tmax), and tended to be higher in females (Figure 21B) compared to males (Figure 21A). Similarly, the mean exposure based on AUCinf was 1979 ± 777 hour·ng / ml and 4741 hour·ng / ml for low-dose males and females, respectively, 4317 ± 1721 hour·ng / ml and 8610 hour·ng / ml for medium-dose males and females, respectively, and 12022 ± 5458 hour·ng / ml and 8305 ± 1607 hour·ng / ml for high-dose males and females, respectively. The mean Cmax and mean AUCinf values increased relatively proportionally with dose (all doses in males, low to medium doses in females), and in females, at low and medium doses, tended to be approximately 2-fold higher (1.7 - 2.4-fold higher) compared to males. The mean t1 / 2 was consistently short across the dose groups, ranging from 0.5 - 0.9 hours.

[0214] Pharmacokinetic parameters in two non-GLP 7-day repeated-dose studies in beagle dogs (SRI number B677-13 for the formulated capsules and SRI number M835-11 for the API delivered by oral gavage) were generally consistent with those of a single-dose GLP study, and 3,4,3-LI(1,2-HOPO) showed no accumulation in plasma after 7 days of dosing. These two pilot studies included an intravenous administration group so that oral bioavailability could be calculated. Oral bioavailability was low, less than 3% and essentially the same in both studies, regardless of whether the dogs received the formulated capsules or 3,4,3-LI(1,2-HOPO) dissolved in PBS without sodium oleate. One male and two female dogs were administered intravenously at 37.6 mg / kg (50 μmol / kg), resulting in peak plasma levels of 115 ± 11 μg / ml, an average AUCinf of 64 h·μg / ml, and a t1 / 2 value of 0.4 h. The volume of distribution was 0.3 L / kg, consistent with a drug that distributes mainly in the extracellular space. Clearance (Cl) was 594 ml / h / kg, and it was shown that plasma clearance could become saturated at higher plasma concentrations. (Example 7) Conclusions of the pharmacokinetic and ADME studies from Examples 5 and 6

[0215] The pharmacokinetics of 3,4,3-LI(1,2-HOPO) were generally comparable across species. The oral bioavailability of formulated and unformulated 3,4,3-LI(1,2-HOPO) was low, less than 3% in dogs, and the bioavailability of total radioactivity from [14C]-3,4,3-LI(1,2-HOPO) in mice and rats was also less than 3%. The formulation of the API with sodium oleate enhanced the exposure parameters by approximately 2- to 3-fold in mice. When administered orally, 3,4,3-LI(1,2-HOPO) was excreted almost completely via the fecal route, either as the unabsorbed parent compound or as metabolites formed either by the liver or in the small intestine. Evaluation of the metabolite profile in rats indicated that the putative major metabolite of [14C]-3,4,3-LI(1,2-HOPO) was feces-specific and accounted for approximately 10% of the administered oral dose. This metabolite may be of biliary origin or is likely the product of metabolism in the intestinal tract, either via a spontaneous degradation process or mediated by the intestinal flora. Thus, the observed low bioavailability of [14C]-3,4,3-LI(1,2-HOPO) is likely due to both in vivo conversion processes and relatively low absorption after oral dosing.

[0216] When administered to rodents via the intravenous route, 3,4,3-LI(1,2-HOPO) was rapidly distributed to the liver and kidneys and excreted via both the renal and biliary routes. The compound was metabolized in the gastrointestinal tract, slightly degraded in gastric juice, and appeared to be stable in plasma.

[0217] Plasma concentrations after oral dosing reached a peak at similar times after dosing (Tmax) in rodents and when using the clinical formulation in dogs (approximately 0.7 hours after dosing and 0.6 - 1.1 hours after dosing in rodents and dogs, respectively). The mean t1 / 2 after oral administration was consistently short across dose groups and species, approximately 1 hour in rats and 0.5 - 0.9 hours in dogs. Cmax, AUC, and oral bioavailability were all approximately 2-fold higher in females than in males in all three species (mice, rats, and dogs), and increased relatively proportionally to the dose. When co-formulated with sodium oleate as a permeation enhancer, radiolabeled [14C]-3,4,3-LI(1,2-HOPO) exhibited increased exposure, which was interpreted as an approximately 2 - 3-fold improvement in oral bioavailability in mice. In beagle dogs administered the clinical formulation of 3,4,3-LI(1,2-HOPO), exposure based on Cmax and AUCinf values increased relatively proportionally to the dose (in males at all doses, in females from low to medium doses), and in females, at low and medium doses, tended to be higher compared to males (1.7 - 2.4-fold higher). The mean t1 / 2 was consistently short across all dose groups, in the range of 0.5 - 0.9 hours.

[0218] Plasma protein binding varied across species and was highest in dogs (95%) when tested at 10 μg / ml, moderate in humans (29%), and lowest in rats (5%). In in vitro human liver microsome experiments, the compound was shown to be relatively stable, and these results are consistent with the relatively low degree of metabolism observed in vivo in rats. 3,4,3-LI(1,2-HOPO) did not inhibit the activities of CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, and thus is unlikely to be a source of drug interaction with other drugs metabolized by these enzymes. (Example 8) GLP Safety Study of Single Oral Dosing in Beagle Dogs

[0219] The clinical formulation was tested in a GLP toxicology and cardiovascular safety pharmacology study of single-dose administration in beagle dogs. The study design is presented in Table 8.1. This study demonstrated the NOAEL of 3,4,3-LI(1,2-HOPO) at 37.5 mg / kg (50 μmol / kg) in dogs after oral capsule administration of the material formulated at 37.5, 75, or 150 mg / kg (50, 100, and 200 μmol / kg).

Table 8

[0220] Based on this study, the MTD in dogs after single oral administration is considered to be higher than 150 mg / kg. All dogs (6 per sex / group, 48 total) survived until scheduled euthanasia on Day 2 or Day 15, and administration of 3,4,3-LI(1,2-HOPO) caused no significant changes in body weight, food consumption, ophthalmic, cardiovascular evaluations, clinical pathology, urine analysis, observations by gross necropsy, or organ weights.

[0221] Clinical observations related to the administration of the test article included diarrhea and vomiting after administration. Specifically, dogs in the medium-dose and high-dose groups experienced mild or severe diarrhea 1 hour to 6 hours after administration on day 1. Diarrhea was not present in the low-dose group. Mild diarrhea was seen in 17% of males and 33% of females in the high-dose group and in 67% of females in the medium-dose group. Severe diarrhea was seen in 33% of males in the high-dose group. By day 2, 8 out of 9 affected dogs had returned to normal, except for 1 female in the high-dose group in which mild diarrhea persisted. Mild to moderate vomiting occurred in 1 to 3 dogs each from the 3,4,3-LI(1,2-HOPO) treatment groups before 2 hours after administration (and in some cases before 1 hour after administration), and did not occur in dogs treated with the vehicle control. Vomiting after dosing occurred in 0, 1, 2, and 3 dogs per group of 12 dogs in the control, low-dose, medium-dose, and high-dose groups, respectively, and thus vomiting was considered dose-dependent. Vomiting after dosing is a common response to oral dosing in dogs, and the single occurrence in the low-dose group is more likely related to the treatment rather than the test article and is not considered a dose-limiting event. The mild vomiting after dosing in this 1 male dog was the only finding present in the low-dose treatment group at 37.5 mg / kg. Similar dose-dependent transient vomiting and diarrhea were observed at approximately 1 hour after dosing in a non-GLP repeated-dose beagle dog study (SRI number B677-13), in which formulated capsules were administered once daily for 7 days at doses of 75.1, 150, or 300 mg / kg (100, 200, and 400 μmol / kg) to 2 dogs / sex. However, loose stools or diarrhea were not observed in Sprague-Dawley rats after oral gavage administration at 400 - 1300 mg / kg (532 - 1732 μmol / kg) for 7 days (SRI number M801-10) or at 7.7 - 76.9 mg / kg (10 - 102 μmol / kg) for 28 days (SRI number M512-07).

[0222] Scattered moderate proximal tubular casts and dilation, moderate interstitial hemorrhage, and mild interstitial hemosiderin pigment deposition in the kidneys were observed in 1 of 3 female dogs in the high-dose group (150 mg / kg) at the time of necropsy on Day 15. Similar kidney histopathological findings were not found in any of the other dogs in the study. Observation of hemosiderin without fibrosis in kidney sections from this one dog was consistent with the kidney findings that occurred within 1 to 3 days prior to necropsy on Day 15. This dog also had corresponding small increases (1.8-fold and 1.3-fold, respectively) in renal function markers BUN and CR on Day 15 compared to pre-test. The timing of the kidney findings in this recovery group of dogs is surprising considering that 3,4,3-LI(1,2-HOPO) was administered only on Day 1. Therefore, it is unclear whether these kidney findings are particularly related to the high-dose administration of 3,4,3-LI(1,2-HOPO). No toxicologically significant histopathological findings were present in any of the dogs at the 2-day time point or in any of the other dogs at the 15-day time point. Similar kidney findings were not present in a non-GLP 7-day repeated-dose dog study (SRI number B677-13) in which 2 dogs / sex received a higher dose of the formulated material for 4 days followed by an equal dose for 3 days and were then necropsied on Day 8. In Sprague-Dawley rat studies, no kidney findings were present after 7-day and 28-day oral gavage dosing (SRI numbers M801-10 and M512-07, respectively).

[0223] Since 3,4,3-LI(1,2-HOPO) is a potent metal chelator, the evaluation of iron serum levels, unsaturated iron binding capacity, total iron binding capacity, magnesium, and ferritin was included in the clinical pathology analysis during the canine safety study. In a GLP study using 3 dogs / sex, none of these parameters changed significantly with statistical significance in the treatment group compared to the control group on Day 2 or Day 15, but the standard deviation was large and the number of dogs per group was small. In a non-GLP repeated dosing pilot study with an even smaller number (2 dogs / sex) and no control group, it was shown that the total iron level in serum increased approximately 2-fold after treatment compared to the pre-treatment level, while the unsaturated iron binding capacity decreased by 16 - 62%, suggesting that the increased iron was bound to transferrin rather than 3,4,3-LI(1,2-HOPO). Therefore, the results obtained from the non-clinical canine studies suggest that serum iron and magnesium levels do not change significantly after administration of 3,4,3-LI(1,2-HOPO), but these parameters are being evaluated in clinical trials. (Example 9) GLP Single Oral Dose GLP Safety from Cardiovascular Evaluation Study in Beagle Dogs

[0224] The cardiovascular parameters in beagle dogs after single oral administration of the formulated capsules were determined in a GLP study.

[0225] Electrocardiograms and blood pressures were evaluated in three male and three female beagle dogs per dose group (0, 37.5, 75, or 150 mg / kg) before the test, 1 hour and 4 hours after a single oral administration of the formulated material, and 7 days later (Table 8.1). There were no electrocardiogram, heart rate, or blood pressure findings attributable to the administration of 3,4,3-LI(1,2-HOPO). Cases of elevated or decreased blood pressure (hypertension or hypotension) in various dogs were considered to be sudden or due to stress, excitement, or difficulty during the recording period and were not considered related to the test article. In summary, no concerns regarding cardiovascular safety arose from the electrocardiogram and blood pressure evaluations in beagle dogs. (Example 10) Enhancement of the permeability of 3,4,3-LI(1,2-HOPO)

[0226] Overview The goal of the analytical study described in Part C of this report was to evaluate the feasibility of enhancing the permeability of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO) using an oral permeation enhancer. The evaluation was performed using an in vitro pK assay based on the Double-Sink (trademark) PAMPA technology developed by pION, Inc.

[0227] Fifteen different permeation enhancers were evaluated for their ability to increase the permeability of 3,4,3-LI(1,2-HOPO) using an in vitro PAMPA assay with an artificial GIT lipid membrane. A significant increase in permeability was observed for one formulation containing 10 mg / mL polysorbate 80 and 1 mg / mL API. All other tested formulations showed no or little improvement in permeability. The formulation containing polysorbate 80 was further evaluated in in vivo studies.

[0228] 1. Purpose of the study The objective of this study was to obtain data that could be used to support the results of the study. This was not conducted in accordance with the regulations of the U.S. Food and Drug Administration (FDA), "Good Laboratory Practice for Nonclinical Laboratory Studies" (GLP) as described in 21 CFR Part 58. However, this study was planned, conducted, recorded, and reported in accordance with standard practices to ensure the quality and completeness of the data.

[0229] 2. Objectives of the Study The objective of this study was to evaluate the feasibility of enhancing the permeability of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO) using oral permeation enhancers. The evaluation was conducted using an in vitro pK assay based on the Double-Sink (trademark) PAMPA technology developed by pION, Inc.

[0230] 3. Experimental Design The permeability enhancement study was conducted in two stages. In the first stage (Table 10.1), 15 formulations were prepared and screened. The second stage (Table 10.2) was conducted to examine the concentration of the permeation enhancer that was found to show enhanced permeability in the first screening. The screening conditions for both stages are listed below. The sample solutions were placed in 20 mL clear scintillation glass vials with polypropylene caps and pulp foil liners during the study and stored wrapped in aluminum foil.

Table 10-1A

Table 10-1B

Table 10-2A

Table 10-2B

[0231] b. Characterization of Samples Visual Observation: For each sample solution, the visual observation consisted of recording the color and transparency.

[0232] pH Recording: The pH of each sample solution prepared for permeability analysis was measured and recorded.

[0233] c. Permeability Assay In vitro PK assay based on the Double-Sink (trademark) PAMPA assay layout:

[0234] The PAMPA Evolution96 (trademark) instrument was used for liquid handling, UV data collection, and result processing. This system consisted of a 96-well Double-Sink PAMPA sandwich with a pre-mounted stir bar. The PAMPA sandwich was formed such that each composite well was divided into two chambers, separated by a 125 μm microfilter disk (pore 0.45 μm), and coated with the Pion GIT-0 lipid mixture. The formulation was suspended in Prisma (trademark) buffer. An artificial membrane separating the two chambers of the permeation system was formed with GIT-0 lipid colored with a filter aid, and simultaneously, a drug-free acceptor sink buffer (ASB, pH 7.4) was placed in the receiving compartment.

[0235] After introducing the formulation into the donor compartment, the PAMPA sandwich was incubated for 15 - 30 minutes or up to 24 hours, and only the UV spectrum on the acceptor side was collected. Calibration for in vivo conditions and agitation of individual wells were provided by Gut-Box™ (Pion Inc.).

[0236] The rate of appearance of the compound in the acceptor compartment of the PAMPA sandwich containing the formulation in the donor compartment was compared with the corresponding rate in the formulation-free system. The ratio of these two rates was reported as the flux ratio. Sample mapping scheme (Tables 10.3 and 10.4):

Table 10-3

Table 10-4

[0237] 5. Results a. Results of the PAMPA assay Observations (appearance and pH of the formulation) and results of PAMPA permeability are summarized in Table 10.5 below for both screening stages. Based on the data obtained from the permeability assay, the GIT lipid coating was stable in the presence of all tested formulations and formulation vehicles, and no leakage was detected. API 3,4,3-LI(1,2-HOPO) showed very low permeability, equivalent to or lower than the permeability level of the reference compound ranitidine.

Table 10-5A

Table 10-5B

Table 10-5C

[0238] For Formulations 1, 2, 4A, 8, 9, 11, 12, and 15, the UV-visible signal in the acceptor compartment was below the detection limit, and the flux ratio could not be determined. Formulations 6 and 7 showed very high permeation rates relative to the corresponding vehicles that completely saturated the UV-visible signal in the acceptor compartment, and signal detection of the API under a strong vehicle background was impossible. Formulations 3, 3A, 4, 5, 5A, 10, 10A, 13, 13A, 13B, 14, 14A, and 14B showed no or little improvement in flow compared to the control API, while Formulation 3B showed a significant improvement in permeability. b. Comparison of Flux Ratio

[0239] The flux ratios obtained for various formulations of 3,4,3-LI(1,2-HOPO) are summarized in Table 10.6 and shown graphically in Figure 22. The only formulation, Formulation 3B, which resulted in a 75-fold increase in permeability, was obtained with 10 mg / mL of polysorbate 80 and 1 mg / mL of API, and the recorded pH was 3.72.

Table 10-6

[0240] 6. Conclusions of Section C Fifteen different permeation enhancers were evaluated for their ability to increase the permeability of 3,4,3-LI(1,2-HOPO) using an in vitro PAMPA assay with an artificial GIT lipid membrane. A significant increase in permeability was observed for one formulation containing 10 mg / mL of polysorbate 80 and 1 mg / mL of API. All other tested formulations showed no or little improvement in permeability. Formulations containing polysorbate 80 will be further evaluated in in vivo studies.

[0241] Analysis of lot ML-11-276 of 3,4,3-LI(1,2-HOPO) was performed for appearance, IR and 1A certificate of analysis was prepared for identification by H-NMR, related compounds by HPLC, HPLC purity, heavy metal content, residual solvent content, water content by Karl Fischer, loss on ignition, and purity.

Table 10-7A

Table 10-7B

Table 10-7C

Table 10-7D

Table 10-7E

Table 10-7F

Table 10-7G

Table 10-7H

Table 10-8A

Table 10-8B

[0242] Overview The goal of the analytical research described in this report was to evaluate the ability of additional oral permeation enhancers to enhance the permeability of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO). The evaluation was performed using an in vitro pK assay based on the Double-Sink (trademark) PAMPA technology developed by pION, Inc.

[0243] In addition to the original 15 types described in Study 12-003-C, 31 different permeation enhancers were evaluated for their ability to increase the permeability of 3,4,3-LI(1,2-HOPO) using an in vitro PAMPA assay with an artificial GIT lipid membrane. Significant increases in permeability were observed in two formulations containing 2-octyl-1-dodecanol and sodium oleate, respectively. All other tested formulations showed no or little improvement in permeability. The formulation containing polysorbate 80 was re-evaluated but was unable to reproduce the previous enhancement results (described in 12-003-C).

[0244] 1. Purpose of the study The purpose of this study was to obtain data that could be used to support the results of the study. This was not conducted in accordance with the regulations of the U.S. Food and Drug Administration (FDA), "Good Laboratory Practice for Nonclinical Laboratory Studies" (GLP) as described in 21 CFR Part 58. However, this study was planned, executed, recorded, and reported in accordance with standard practices to ensure the quality and completeness of the data.

[0245] 2. Objectives of the Study The objective of this study was to evaluate the ability of additional oral absorption enhancers to enhance the permeability of the active pharmaceutical ingredient 3,4,3-LI(1,2-HOPO). The evaluation was performed using an in vitro pK assay based on the Double-Sink (trademark) PAMPA technology developed by pION, Inc. This is a follow-up study to LBNL No. 12-003-C that initially tested 15 oral absorption enhancers.

[0246] 3. Experimental Design The permeability enhancement study was conducted in two additional stages (Stages 3 and 4, following Stages 1 and 2 described in Example 10). In Stage 3, 32 formulations were prepared and screened (31 new formulations and one repeat of the most successful formulation obtained in Example 10). In Stage 4, the concentrations of the absorption enhancers that were seen to show permeability enhancement in the first three screenings were scrutinized and performed to verify reproducibility. The screening conditions for both Stages 3 and 4 are listed in Table 11.1 and Table 11.2 below, respectively. The sample solutions were placed in 20 mL clear scintillation glass vials with polypropylene caps and pulp foil liners and stored wrapped in aluminum foil during the study.

Table 11-1A

Table 11-1B

Table 11-1C

Table 11-2

[0247] 5. Results a. Results of the PAMPA assay Summarize the observations (appearance and pH of the formulations) and the results of PAMPA permeability for screening stages 1 and 2 in Table 11.3, and for screening stages 3 and 4 in Table 11.4. Based on the data obtained from the permeability assay, the GIT lipid coating was stable in the presence of all the formulations and formulation vehicles tested, and no leakage was detected. API 3,4,3-LI(1,2-HOPO) showed very low permeability and was equal to or lower than the permeability level of the reference compound ranitidine.

Table 11-3A

Table 11-3B

Table 11-3C

Table 11-4A

Table 11-4B

Table 11-4C

Table 11-4D

[0248] For Formulations 17, 18, 20, 22, 23, 24, 27, 28, 29, 40, 42, 43, 44, 45, and 3B, the UV-visible signal in the acceptor compartment was below the detection limit, and the flux ratio could not be determined. Formulations 32 and 38 showed very high permeation rates relative to the corresponding vehicle that completely saturated the UV-visible signal in the acceptor compartment, and signal detection of the API under a strong vehicle background was not possible. The significant improvement in permeability previously seen for Formulation 3B was not reproducible in subsequent iterations. A significant and reproducible improvement was observed for Formulation 26, and a significant improvement was also observed for Formulation 37. Formulations 16, 19, 21, 25, 30, 31, 33, 34, 35, and 36 showed no or little improvement in flow compared to the control API.

[0249] b. Comparison of Flux Ratio The flux ratios obtained for the various newly tested formulations of 3,4,3-LI(1,2-HOPO) are summarized in Table 11.5. In the iteration of Formulation 3B, the initial 75-fold increase in permeability was not reproduced. However, Formulation 26 resulted in a reproducible enhancement, obtained with 2.50 mg / mL sodium oleate and 1 mg / mL API, and the recorded pH was 8.81.

Table 11-5A

Table 11-5B

Table 11-5C

[0250] 6. Conclusions Thirty-one additional permeation enhancers were evaluated for their ability to increase the permeability of 3,4,3-LI(1,2-HOPO) using an in vitro PAMPA assay with an artificial GIT lipid membrane. The significant increase in permeability initially observed for one formulation containing 10 mg / mL polysorbate 80 and 1 mg / mL API was not reproducible. Most of the other tested formulations showed little or no improvement in permeability, although improvement was observed for formulations containing 2.50 mg / mL sodium oleate or 2-octyl-1-dodecanol. The formulations containing sodium oleate or 2-octyl-1-dodecanol were further evaluated in vivo.

[0251] Analysis of lot ML-11-276 of 3,4,3-LI(1,2-HOPO) was performed, and a certificate of analysis was prepared for appearance, identification by IR and 1 1H-NMR, related compounds by HPLC, HPLC purity, heavy metal content, residual solvent content, water content by Karl Fischer, loss on ignition, and purity.

Table 11-6A

Table 11-6B

Table 11-6C

[0252] The feasibility of developing an oral formulation of 3,4,3-LI(1,2-HOPO) was evaluated.

[0253] Four oral dosage forms were investigated: (i) bottled powder, (ii) dispersible / soluble granules, (iii) chewable tablets, and (iv) conventional immediate-release tablets. Based on the studies conducted, nine formulation prototypes showing immediate drug release behavior and required physical properties were identified and selected for API verification, gastric dissolution, and testing of related substances by a specified liquid chromatography method. Of these selected compositions, two were bottled powder formulations, two were granule formulations, three were chewable tablet formulations, and two were conventional tablet formulations. The respective compositions of these prototype formulations are summarized and presented below. All assays confirmed that these prototypes are suitable for further development.

Table 12

[0254] It should be understood that the present invention is described in conjunction with its preferred specific embodiments, but the foregoing description is intended to be illustrative and not intended to limit the scope of the present invention. Other aspects, advantages, and modifications within the scope of the present invention are expected to be apparent to those skilled in the art to which the present invention pertains.

[0255] All patents, patent applications, and publications referred to herein are hereby incorporated by reference in their entirety.

Claims

1. A pharmaceutical composition for use in the treatment of a subject with heavy metal exposure, wherein the pharmaceutical composition comprises a 1,2-HOPO chelating agent in an amount of about 100 to about 1500 mg, which is 3,4,3-LI-1,2-HOPO, sodium oleate, and wherein the subject has an excess of heavy metals and the pharmaceutical composition is effective to remove, clear and / or reduce the excess heavy metals from the subject in vitro.

2. The pharmaceutical composition for use according to claim 1, wherein the subject has been exposed to, is in contact with or is contaminated by one or more metals from the actinide or lanthanide group or a mixture thereof.

3. The pharmaceutical composition for use according to claim 2, wherein the administration results in the in vitro removal, clearance and / or reduction of an excess of actinides, lanthanides or both from one or more systems and / or organs of the subject.

4. The pharmaceutical composition for use according to claim 1, wherein the heavy metals comprise one or more metals from the lanthanide or actinide group or a mixture thereof.

5. The pharmaceutical composition for use according to claim 1, wherein the heavy metals comprise lead, tin, yttrium, scandium and / or cadmium.

6. The pharmaceutical composition for use according to claim 1, wherein the sodium oleate is present in an amount of about 70 mg to about 130 mg.

7. The pharmaceutical composition for use according to claim 1, wherein the sodium oleate is present in 8 to 12% of the total weight of the pharmaceutical composition.

8. The pharmaceutical composition for use according to claim 1, wherein the sodium oleate is about 11% of the total weight of the pharmaceutical composition.

9. The pharmaceutical composition for use according to claim 1, wherein the 3,4,3-LI-1,2-HOPO is present in an amount of 300 mg to 1500 mg.

10. The pharmaceutical composition for use according to claim 1, wherein the 3,4,3-LI-1,2-HOPO is present in an amount of 400 mg to 1200 mg.

11. The pharmaceutical composition for use according to claim 1, wherein the 3,4,3-LI-1,2-HOPO is present in an amount of 100 mg to 300 mg.

12. The pharmaceutical composition for use according to claim 1, wherein the 3,4,3-LI-1,2-HOPO is present in an amount of 600 mg.

13. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is a powder.

14. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is a chewable tablet.

15. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is an immediate-release tablet.

16. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition is in one or more orally dispersible / soluble granules.

17. The pharmaceutical composition for use according to claim 2, wherein the administration is carried out 24 hours after exposure to, contact with, and / or contamination by one or more metals.

18. The pharmaceutical composition for use according to claim 1, wherein the pharmaceutical composition comprises one or more additional components.

19. The pharmaceutical composition for use according to claim 18, wherein the one or more additional components include microcrystalline cellulose, carboxymethyl cellulose, croscarmellose sodium, guar gum, lactose monohydrate, hypromellose, magnesium stearate, povidone, crospovidone, mannitol, colloidal silicon dioxide, compressed sugar, pregelatinized starch, sodium starch glycolate, hydrogenated vegetable oil (type 1), and polysorbate 80.

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