Novel Liquid Formulation for Iron Chelation

Abstract

An iron chelate preparation containing a chelating agent (such as EDTA and its salts) and methylsulfonylmethane (MSM) is provided. The preparation can be an immediately drinkable beverage, a hydration drip, or any liquid that can be administered systemically to a subject. Using the disclosed composition, a preparation and method for treating RNA viruses including SARS-CoV-2 are provided. A preparation and method for inhibiting ferroptosis and diseases and conditions affected by ferroptosis are provided. Diseases and conditions affected by ferroptosis include cancer, aging, inflammation, and the like.

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims the priority of U.S. Provisional Patent Application No. 63 / 305,099, titled "Novel Liquid Formulation for Iron Chelation", filed on January 31, 2022, the content of which is incorporated herein by reference in its entirety.

[0002] (Field of the Invention) The present invention generally relates to the field of agents for treating ferroptotic cell death in the case of cancer, aging, viral infection, and other ferroptosis - related pathologies. More specifically, it relates to consumable formulations containing chelating agents and transport - promoting agents suitable for chelating intracellular iron. In particular, the present invention relates to a ready - to - drink beverage formulation containing a transport - promoting agent and a chelating agent. In an exemplary embodiment, it relates to a potable liquid composition containing MSM as a transport - promoting agent and a chelating agent generally recognized as safe (GRAS).

Background Art

[0003] Iron (Fe) plays a major role in human diseases. Certain viruses (RNA viruses) that can cause fatal infections in humans are iron - dependent for replication within the human body. For example, treatment with the iron - chelating agent deferiprone has been shown to extend the survival period of patients with acquired immunodeficiency syndrome (AIDS). Iron overload has been found to cause iron deficiency, which is the cause of many human diseases. Ferroptosis is thought to be related to the debilitating aspects of the aging process. Therefore, restricting iron is a promising adjunctive strategy for treating viral infections, either through oral ingestion or intravenous injection of iron - chelating agents, or through manipulation of major iron - regulatory factors.

[0004] The coronavirus is a type of single-stranded RNA virus with an envelope. In the past few decades, two highly pathogenic strains of coronavirus, Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV), have been identified in humans. These viruses have been found to cause severe and sometimes fatal respiratory diseases. In December 2019, a new strain of coronavirus (SARS-CoV-2) caused Coronavirus Disease 2019 (Covid-19), and on March 11, 2020, the WHO declared a pandemic. Common symptoms of Covid-19 infection include respiratory symptoms, fever, cough, shortness of breath, and difficulty breathing. In more severe cases, the infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and death. As of January 2022, more than 373 million infections and more than 5.66 million deaths have been reported worldwide.

[0005] Due to the rapid mass vaccination and mask-wearing protocols introduced worldwide, the severity of the illness may have been reduced in many patients, but the virus has shown the ability to regularly mutate into new variants that are highly lethal (such as Delta), highly infectious (such as Omicron), or have the ability to evade immunity generated by vaccination or prior infections with variants. However, at present, there is no specific treatment, and patients have no choice but to rely on general treatments and supportive therapies such as oxygen supply and broad-spectrum antiviral drugs. Remdesivir, a new nucleotide analog prodrug being developed as a treatment for Ebola virus and Middle East Respiratory Syndrome (MERS) disease, has been reported to relieve the symptoms of pneumonia caused by COVID-19 infection. Given that the pandemic is occurring on a global scale, there is an urgent need for drugs and methods to inhibit or block the replication of the SARS-CoV-2 virus, and it is necessary to make such drugs and methods widely and easily accessible to many people around the world who may be infected with the SARS-CoV-2 virus and its variants.

[0006] Iron (Fe) has been demonstrated to play two major roles in the novel coronavirus infection and the severity of its symptoms. First, all coronaviruses (including SARS-Cov2) require iron for replication (Shi ST, et al. CTMI (2005) 287:95-131; Liu W, et al. Current Clinical Microbiology Reports https: / / doi.org / 10.1007 / s40588-020-00140-w (2020); Habib HM, et al. Biomedicine & Pharmacotherapy 136 (2021) 111228). RNA viruses have evolved to be highly dependent on iron for replication, and replication is slower in iron-deficient situations (Menshawey R, et al. Egyptian Journal of Medical Human Genetics (2020) 21:75). Increased intracellular iron efflux due to increased expression of the iron exporter ferroportin also shows an antiviral effect against the human immunodeficiency virus (HIV). (Liu 2020). Second, COVID-19 patients are often troubled by iron overload, which results in iron deficiency and damages multiple organs. (Chen X, et al. J. Exp. Med. 2021 Vol. 218 No. 6 e20210518 (2021); Yang M, et al. Yang and Lai Cell Death Discovery (2020) 6:130; Edeas M, et al. International Journal of Infectious Diseases 97 (2020) 303-305)

[0007] The onset of COVID-19 is associated with hyperferritinemia and changes in iron homeostasis. Hyperferritinemia has been described as a major feature that predicts a high significance in the increased risk of death due to novel coronavirus infection. (Mehta P, et al., Lancet 2020;395(March(10229)):1033-4). These studies demonstrated that the serum ferritin levels of those who could not survive COVID-19 exceeded twice the level of survivors.

[0008] Ferroptosis is a type of regulated necrosis caused by a combination of iron toxicity, lipid peroxidation, and cell membrane damage. This is a programmed cell death pathway that depends on intracellular iron and not on other metals. Non-apoptotic cell death can promote the selective elimination of some tumor cells or be activated in certain pathological conditions. Tumorigenic RAS-selective lethal small molecule erastin was found to cause a unique iron-dependent non-apoptotic cell death. (Dixon SJ, et al. Cell. May 25, 2012; 149(5): 1060-1072). In 2012, Dixon coined the term ferroptosis for an iron-dependent non-apoptotic cell death mode characterized by the accumulation of lipid reactive oxygen species (ROS). This is mainly caused by an increase in redox imbalance, but is morphologically, biochemically, and genetically different from other known cell death patterns such as apoptosis, necrosis, and autophagy.

[0009] Ferroptotic cells and their efflux contents shape innate and adaptive immunity in health and disease. Excessive or insufficient ferroptotic cell death is thought to be increasingly involved in many physiological and pathophysiological processes in combination with dysregulation of the immune response.

[0010] In senile and degenerative diseases, the iron concentration in the brain inevitably increases. Oxidative stress caused by excessive iron is associated with carcinogenesis. Acute kidney injury (AKI), formerly known as acute renal failure (ARF), is a common and severe disease caused by multiple factors such as ischemia, nephrotoxic drugs, and urinary tract obstruction. Although various molecular mechanisms have been proposed to induce or exacerbate AKI, ROS-induced kidney injury is considered one of the main mediators. AKI occurs in approximately 5% of inpatients and 30% of critically ill patients, with high morbidity and mortality. Furthermore, studies have shown that AKI increases the potential risk of chronic kidney disease and end-stage renal disease in patients. In addition to blood purification, there have been few therapeutic methods that have made great progress in the prevention of AKI. Therefore, new targets or better regimens are still urgently needed to prevent AKI and promote adaptive repair after AKI occurs. Multiple studies have suggested that ferroptosis is a promising therapeutic target, especially in diseases mainly caused by renal tubular necrosis. (Linkermann A. et al. Journal of the American Society of Nephrology. 2014;25(12):2689-2701.)

[0011] It is widely recognized that chelating iron improves and / or prevents ferroptosis. Unfortunately, the chelating agents proposed in the literature are only deferoxamine (DESFERAL®), which is only approved for intravenous and subcutaneous administration, 2,2'-bipyridyl (2,2'-dipyridine), a highly toxic substance that shows effects in cell culture, and ciclopirox olamine (LOPROX®), a chelating fungicide for topical use that is not approved for systemic use. All of these have safety problems, are not suitable for long-term use to prevent ongoing ferroptosis, and have not been evaluated at non-toxic doses for long-term use. Ferroptosis is one of the promising therapeutic targets, especially in diseases mainly caused by renal tubular necrosis such as ischemic, cisplatin nephrotoxicity, and rhabdomyolysis-induced AKI.

[0012] Generally, lipid peroxidation inhibitors such as lysyl oxidase inhibitors, ferrostatin-1, and liproxstatin-1 are used to inhibit ferroptosis. In some of these models, iron chelators have been studied long before ferroptosis is detected. However, compounds such as deferoxamine have not been used in the clinical setting, despite having significant effects in in vitro experiments in renal tubules. Antioxidants and iron chelators (such as vitamin E and deferoxamine) have also been observed to inhibit ferroptosis by reducing iron availability, but this has only been observed in in vitro experiments and rodent models. Chronic administration of deferoxamine has been reported to have multiple side effects, such as acute respiratory distress syndrome, visual impairment, and enhanced infection with Yersinia enterocolitica.

[0013] When used in patients without iron overload, deferoxamine can cause iron deficiency and may reduce ferritin concentration. In the case of single-dose administration, the most severe side effects are flushing, erythema, tachycardia, urticaria, and hypotension caused by rapid administration of deferoxamine. Deferoxamine is only approved for intramuscular and intravenous administration in cases of acute iron poisoning.

[0014] There is a need to reduce and treat diseases, symptoms (including aging), and viral infections that may be caused by iron deficiency by systemically administering iron chelators to subjects (humans or animals). Iron chelators commonly used in foods include disodium EDTA, calcium disodium EDTA, and metaphosphate. These may have the potential to be used for oral administration, but generally are not absorbed into the body by oral administration.

[0015] Therefore, there is a need for a novel formulation comprising an iron chelating agent that can be administered systemically and one or more penetration enhancers that enable the chelating agent to be absorbed into the body. In particular, during a pandemic caused by a rapidly mutating RNA virus, there is a need for a desirable formulation that can suppress viral replication in a form that can be easily used to reduce the iron concentration in a subject and thereby suppress viral replication and the viral load.

SUMMARY OF THE INVENTION

[0016] The present invention provides a formulation that can be used to control the iron level in the body by periodically replenishing an iron chelating agent to a subject. Diseases and conditions associated with high iron levels, such as viral infections and hemosiderosis, are thereby regulated. In one embodiment, the formulation is a potable liquid such as a drinkable beverage. The novel formulations disclosed herein are useful for controlling the viral load in an infected subject and are also useful for ferroptosis-mediated diseases and conditions such as cancer and aging.

[0017] Formulations containing a transport enhancer (such as MSM) and a chelating agent (such as EDTA) for topical application to the eye, teeth, and skin surface have been previously disclosed and patented by the inventors. (WO 2013 / 166459 by Bhushan et al.; WO 2014 / 100775 by Bhushan et al.; U.S. Patent No. 9,616,008 by Bhushan et al.).

[0018] The present invention provides a formulation comprising a chelating agent suitable for long-term ingestion or a salt thereof, a penetration enhancer that is methylsulfonylmethane (MSM), one or more inert excipients, and a liquid vehicle or carrier, wherein the chelating agent and the penetration enhancer are present in proportions effective to maintain the homeostasis of the iron level in the body when ingested in normal doses, the proportion of the chelating agent is about 0.0001%. to 15%, and the proportion of the penetration enhancer in the composition is about 0.0001% to 30% by weight, respectively.

[0019] In one aspect of the present invention, the formulation is in the form of a concentrate, a liquid, a soluble solid, a foaming tablet, a pill, or a form that can be easily reconstituted.

[0020] In one aspect of the present invention, the formulation is in a liquid form that can be administered by a route selected from the group consisting of oral, intranasal, inhalation, intravenous, intramuscular, transdermal, topical, rectal, vaginal, buccal, injection, sublingual, or combinations thereof. In certain aspects, the formulation includes a rehydration drip further comprising electrolytes, vitamins, or other nutrients.

[0021] In one aspect of the present invention, iron homeostasis is sufficient to inhibit the replication of RNA viruses such as coronavirus, retrovirus, HIV-1, SARS-CoV-2, MERS, SARS, influenza, HTLV-I, HTLV-II, etc. in human or animal subjects.

[0022] In one aspect of the present invention, iron homeostasis is sufficient to inhibit ferroptosis and ferroptotic cell death.

[0023] In certain aspects of the present invention, inhibited ferroptosis is associated with pathological diseases or conditions in a subject, including diseases or conditions associated with organs selected from the heart, central nervous system, liver, gastrointestinal organs, lungs, kidneys, and pancreas.

[0024] In certain aspects of the present invention, inhibited ferroptosis is associated with cancer, aging, inflammation, hearing loss, neurodegenerative diseases, and diseases associated with I / R (ischemia-reperfusion) injury.

[0025] In one aspect of the present invention, ferroptosis is reduced by inhibiting the accumulation of iron-dependent lipid reactive oxygen species (ROS).

[0026] In one aspect of the present invention, iron homeostasis is maintained by chelating iron bound to a heme-containing protein selected from hemoglobin, myoglobin, and neuroglobin.

[0027] This method involves administering to a subject an effective amount of a formulation comprising a therapeutically effective amount of a chelating agent and an effective transport-promoting amount of a transport promoter having the formula (I).

Chemical formula

[0028] Examples of the transport promoter include methylsulfonylmethane (MSM; also called methylsulfone, dimethylsulfone, DMSO 2 ).

[0029] The chelating agent can be selected from ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), cyclohexanediaminetetraacetic acid (CDTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylenephosphonic acid (ArPA), citric acid, acetic acid, phosphoric acid, pyrophosphate, metaphosphate, malic acid polymer, etc., and their acceptable salts, and any combination thereof.

[0030] This specification discloses a method for treating or alleviating Covid-19 by inhibiting the replication of SARS-CoV2.

[0031] This specification discloses a method for preventing, treating or alleviating diseases or symptoms associated with high intracellular iron levels, hyperferritinemia and ferroptosis.

[0032] These aspects and other aspects will become apparent from the following description of the preferred embodiments in conjunction with the following drawings, but their variations and modifications may be affected without departing from the spirit and scope of the novel concept of the present disclosure.

Brief Description of the Drawings

[0033] The following drawings form part of this specification and are included to further demonstrate certain aspects of the present invention. By referring to one or more of these drawings in combination with the detailed description of the specific embodiments presented herein, the invention of the present invention can be better understood.

[0034] The patent or application file includes at least one drawing created in color. A copy of the color drawing of this patent or patent application publication will be provided by the Patent Office if requested and the necessary fee is paid.

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Mode for Carrying Out the Invention

[0043] The terms used in this specification generally have their ordinary meanings in the context of the relevant art, the context of the present invention, and the particular context in which each term is used. Specific terms used to describe the present invention are explained below or elsewhere in this specification to provide additional guidance to practitioners regarding the description of the present invention. For convenience, certain terms may be highlighted using italics, quotation marks, etc. The use of highlighting does not affect the scope and meaning of the term. Whether highlighted or not, the scope and meaning of the term are the same in the same context. It will be understood that the same thing can be expressed in multiple ways. Thus, alternative languages or synonyms may be used for one or more of the terms described herein, and no special meaning is placed on whether the term is described or explained in detail herein. Synonyms for specific terms are provided. The listing of one or more synonyms does not exclude the use of other synonyms. The use of any example in this specification (including examples of any term discussed in this specification) is for illustrative purposes only and does not limit the scope and meaning of the present invention or any term exemplified. Similarly, the present invention is not limited to the various embodiments described herein.

[0044] When a range of values is provided, unless the context clearly dictates otherwise, each intervening value between the upper and lower limits of that range, and any other defined value or intervening value within that defined range, down to the one-tenth unit of the lower limit value, is understood to be included in the present invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are included in the present invention subject to any particular exclusions described within the stated range. When one or both of the limiting values are included in the stated range, ranges excluding one or both of the included limiting values are also included in the present invention.

[0045] When referring to pharmaceutical ingredients, the terms used (e.g., "agent") are intended to include not only specific molecular entities but also their pharmaceutically acceptable analogs (including but not limited to salts, esters, amides, prodrugs, conjugates, active metabolites, and other such derivatives, analogs, and related compounds).

[0046] As used herein, the terms "treat" and "treatment" refer to administering a drug or formulation to a clinically symptomatic human or animal suffering from an adverse condition, disorder, or disease to reduce the severity and / or frequency of symptoms, eliminate the symptoms and / or their underlying cause, and / or promote improvement or repair of the damage. The terms "prevent" and "prevention" refer to administering a drug or composition to a clinically asymptomatic individual who is susceptible to a particular adverse condition, disorder, or disease, and thus relate to preventing the occurrence of symptoms and / or their underlying cause. Unless expressly or implicitly provided otherwise herein, when the term "treat" (or "treating") is used without reference to the possibility of prevention, prevention is also intended to be included, and a "method for treating gingivitis" is construed to include a "method for preventing gingivitis".

[0047] "Optional" or "optionally present" - in the case of an "optional substituent" or "optionally present additive", it means that the component (such as a substituent or additive) described below may or may not be present, and the description includes both the case where the component is present and the case where it is not present.

[0048] "Pharmaceutically acceptable" means a material that is not biologically or otherwise undesirable, e.g., the material can be incorporated into the formulations of the present invention without causing undesirable biological effects or interacting in a deleterious manner with any of the other components of the dosage form formulation. However, when the term "pharmaceutically acceptable" is used to refer to a pharmaceutical additive, it is implicitly indicated that the additive meets the required criteria of toxicity and manufacturing tests and / or is included in the Inactive Ingredients Guide prepared by the US Food and Drug Administration. As will be explained in more detail below, "pharmacologically active" (or simply "active") in a "pharmacologically active" derivative or analog refers to a derivative or analog having the same type of pharmacological activity as the parent drug. As used herein, the terms "treat" and "treatment" refer to the reduction in the severity and / or frequency of symptoms, the elimination of symptoms and / or the underlying cause, the prevention of the occurrence of symptoms and / or their underlying cause, and the improvement or repair of an undesirable condition or injury. Thus, for example, "treating" a subject includes preventing a deleterious condition in a susceptible individual and clinically treating an individual with symptoms by causing suppression or regression of the condition. The term "chelating agent" (or "active agent") refers to any compound, complex, or composition that exhibits a desired effect in a biological context, i.e., when administered to a subject or introduced into cells or tissues in vitro. This term includes pharmaceutically acceptable derivatives of the active agents specifically mentioned herein, including but not limited to salts, esters, amides, prodrugs, active metabolites, isomers, analogs, crystalline forms, hydrates, etc. When the term "chelating agent" is used or a specific chelating agent is specifically identified, it is understood that not only the agent itself but also its pharmaceutically acceptable salts, esters, amides, prodrugs, active metabolites, isomers, analogs, etc. are intended.

[0049] The "effective" or "therapeutically effective" amount of an active agent means an amount of the agent that is non-toxic but sufficient to produce a beneficial effect. The amount of an "effective" active agent will vary for each subject depending on, among other things, the individual's age and general condition, and the particular active agent. Unless otherwise stated, as used herein, the term "therapeutically effective amount" is intended to include an amount effective for the prevention and / or amelioration of a deleterious condition, i.e., in addition to an amount effective for the treatment of a deleterious condition, an amount effective for the prevention and / or amelioration of a deleterious condition.

[0050] As will be apparent to those skilled in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features that can be readily separated from or combined with any of the features of some other embodiments without departing from the scope or spirit of the present invention. Any of the recited methods can be performed in the order of the recited events or in any other logically possible order.

[0051] Unless otherwise specified, the present invention is not limited to specific formulation ingredients, administration methods, chelating agents, manufacturing processes, etc., and can be modified.

[0052] 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 pertains. In case of conflict, the present document, including definitions, will control.

[0053] The Role of Iron in Coronavirus Disease 2019 Iron overload is increasingly suspected as a factor contributing to the onset of COVID-19. In fact, some of the symptoms of COVID-19, such as inflammation, hypercoagulability, hyperferritinemia, and immune dysfunction, are reminiscent of iron overload. Iron is essential for all living cells, but free unbound iron generated by iron dysregulation or excess is highly reactive and potentially toxic because it is involved in the production of reactive oxygen species (ROS). ROS react with cellular lipids, nucleic acids, and proteins to cause damage, resulting in the activation of acute or chronic inflammatory processes associated with multiple clinical symptoms.

[0054] COVID-19 presents itself as a number of complications and physiological and biochemical changes. These include, but are not limited to, acute respiratory distress syndrome (ARDS), high concentrations of inflammatory-induced CD4 T cells and cytotoxic granule CD8 T cells, massive cytokine release (cytokine storm), elevated coagulation status, hemoglobin damage, and dysregulation of iron homeostasis, including iron overload. (Habib, 2021).

[0055] After circulating worldwide for 20 months, the basal lineages of SARS-CoV-2 have been almost completely replaced by derived mutant lineages. These lineages are classified by the WHO as variants of concern (VOC) or variants of interest (VOI) based on genetic, phenotypic, and epidemiological differences. Despite having a certain degree of proofreading ability (a relatively rare function for an RNA virus), SARS-CoV-2 accumulates approximately 24-25 substitutions per year. A rigorous quantification of the evolutionary process shows that the observed success of the mutant viruses is the result of adaptive evolution rather than neutral evolution. (Kissler KE et al., preprint accessed January 28, 2022: https: / / doi.org / 10.1101 / 2021.09.11.459844).

[0056] Iron plays two roles in COVID-19 and the severity of its symptoms. (a) All coronaviruses (including SARS, MERS, SARS-CoV-2) are iron-dependent for replication (Shi 2005, Liu 2020, Habib 2021), and replication is slower in an iron-deficient environment (Menshawey 2020). (b) COVID patients are often troubled by iron overload, resulting in iron deficiency and damage to multiple organs. (Chen 2021, Yang 2020, Edeas 2020).

[0057] The main storage depots of iron in the body are cells including hemoglobin, myoglobin, and neuroglobin. Iron is an essential element for blood production. More than 70 percent of the iron in the body is present as a heme-containing protein called hemoglobin in red blood cells, as a heme-containing protein called myoglobin in muscle cells, and as a heme-containing protein called neuroglobin in nerve cells. Approximately 6 percent of the iron in the body is a component of specific proteins essential for respiration and energy metabolism and is also a component of enzymes involved in the synthesis of collagen and some neurotransmitters. Iron is also necessary for proper immune function. Approximately 20-25 percent of the iron in the body is stored as ferritin in cells and circulating blood.

[0058] For the host, iron is an essential trace element required for many basic enzymatic and non-enzymatic reactions, as well as diverse physiological processes, including mitochondrial functions such as ATP generation, DNA / RNA synthesis and repair, and cell survival / apoptosis (Khodour Y, et al., Enzymes. 2019;45:225-56).

[0059] The main route by which SARS-CoV-2 invades cells is by attaching to angiotensin-converting enzyme 2 (ACE2) that adheres to the outer surface of the cell membranes of cells in the lungs, arteries, heart, kidneys, and intestines. When the spike S1 proteins of SARS-CoV and SARS-CoV-2 bind to the ACE2 enzyme domain on the cell surface, endocytosis occurs, and both the virus and the enzyme migrate to the endosome inside the cell. Liu et al. suggest that the strong affinity between SARS-CoV-2 and the human ACE2 molecule indicates that an important pathogenic molecular stage of COVID-19 is to attack ACE2-positive cells. More than 80% of the ACE2 receptors are expressed in a small population of type II alveolar cells (AT2).

[0060] Iron in the body is very strictly regulated. Sufficient intracellular iron levels support the replication of coronaviruses, while intracellular iron deficiency interferes with viral transcription, translation, assembly, and exocytosis, weakening the replication process. The main storage depots of iron in the body are cells containing heme-containing proteins such as hemoglobin, myoglobin, and neuroglobin. Iron is an essential element for blood production.

[0061] Since coronaviruses require iron to replicate, their replication is promoted when they infect iron-containing cells. However, for the Covid virus to invade cells, the cells need to have the ACE2 receptor. There is no ACE2 receptor on the surface of red blood cells that have hemoglobin. There may be another route on the surface of red blood cells (RBC) that can interact with the S1 spike protein of SARS-CoV2. One such route is the interaction with the RBC Band3 surface protein. (Cosic I, et al., Appl. Sci. 2020, 10, 4053; doi:10.3390 / app10114053). However, the virus also infects other iron-rich cells that contain the ACE2 surface receptor, such as muscle cells (myoglobin) and nerve cells (neuroglobin).

[0062] Hemoglobin, myoglobin, and neuroglobin lose their ability to bind oxygen, impeding the supply of oxygen to major organs and causing rapid multiple organ failure. Furthermore, the free iron released into the circulation can cause iron overload and potentially inflict oxidative damage on the lungs and other organs. Iron overload can also trigger inflammation and immune dysfunction. These determine an increase in the absorption and storage of iron into iron-binding proteins. In fact, this idea is supported by the elevated levels of ferritin (an iron storage molecule in the body) in the blood of COVID-19 patients. The increased iron load raises blood viscosity and leads to recurrent and widespread thrombosis in the systemic and microcirculation, which can cause unexpected deterioration and death.

[0063] The iron-dependence of virus replication and the regulation of host iron metabolism by RNA viruses indicate the importance of cellular iron homeostasis in the viral life cycle and suggest the usefulness of iron chelation strategies in the treatment of viral infections. One strategy is to directly deplete iron with chelating agents that have a strong and selective affinity for iron ions. Some iron chelating agents such as deferoxamine (DFO, DESFERAL®), deferiprone (DFP, FERRIPROX®), and deferasirox (ICL670, EXJADE®) have been approved for clinical use by the US Food and Drug Administration. Iron chelating agents can bind to free iron or remove iron from iron-containing proteins. Deferasirox is a membrane-permeable iron chelating agent and the first oral drug approved by the US Food and Drug Administration (FDA) as a treatment for chronic iron overload in the body caused by multiple blood transfusions. Treatment with high doses of DFP has been shown to extend the survival of AIDS patients after HIV-1 infection. ciclopirox is a synthetic broad-spectrum antifungal drug that binds to trivalent cations such as Fe 3+ is. Dexrazoxane is a cyclic derivative of EDTA that easily penetrates cell membranes. Baicalein is a flavonoid extracted from Scutellaria baicalensis Georgi and has free 5,6,7-hydroxyl groups that form complexes with iron in a 1:1 stoichiometry.

[0064] Iron chelating drugs can bind to free iron, but they can also remove iron from iron-containing proteins, so iron chelation may have an anti-ferritin effect. In fact, deferoxamine promotes the degradation of ferritin by lysosomes. Iron chelation therapy may play an important role in the control and treatment of COVID-19, but the approved iron chelating agents are not suitable for long-term use. The most commonly used approved iron chelating agent (deferoxamine, DFO) has the drawback of degrading ferritin (De Domenico I, et al., BLOOD, November 12, 2009, Vol. 114, No. 20 2009; Abobaker A, European Journal of Clinical Pharmacology (2021) 77:267-268), which may worsen the symptoms. However, food-grade chelating agents, especially EDTA, do not have such an impact on ferritin.

[0065] Ferroptosis The upstream inducers of ferroptosis are divided into two categories (biological vs. chemical), activating two major pathways (exogenous / transporter pathway and endogenous / enzyme pathway). Ferroptosis does not have the typical morphological features of necrosis such as swelling of the cytoplasm and organelles and rupture of the cell membrane, nor does it have the characteristics of conventional apoptosis such as cell shrinkage, chromatin condensation, formation of apoptotic bodies, and breakdown of the cytoskeleton. In contrast to autophagy, classical closed double-membrane structures (autophagosomes) are not formed in ferroptosis. Morphologically, ferroptosis mainly appears as a distinct shrinkage of mitochondria, accompanied by an increase in membrane density and a decrease or disappearance of mitochondrial cristae, which is a process different from other forms of cell death.

[0066] According to current knowledge, ferroptosis has been shown to occur during various pathophysiological processes of the body, such as neurodegenerative diseases, antiviral immune responses, atherosclerosis, acute kidney injury, diabetes, and ischemia-reperfusion injury. (Oxid Med Cell Longev. 2019; 2019:8010614; published online October 31, 2019. Ferroptosis is characterized by the accumulation of membrane lipid peroxidation products and the consumption of polyunsaturated fatty acids in cell membranes. This type of cell death can be induced by specific small molecules such as elastin and RAS-selective lethal 3 (RSL3). Ferroptosis has been reported to be involved in various pathological processes of diseases in the brain, kidney, liver, and heart. (Sheng X., et al. Physical Chemistry Chemical Physics.)

[0067] Depletion of glutathione (L-glutamyl-L-cysteinyl-glycine [GSH]) can lead to the accumulation of iron-dependent reactive oxygen species (ROS), particularly lipid ROS, which may be sufficient to kill cells by itself. (Dickinson, 2012). Iron metabolism and lipid peroxidation signaling are considered to be central mediators of ferroptosis. Circulating iron exists in the form of trivalent iron (Fe3+) bound to transferrin, thereby reducing the level of free iron. Excessive iron may generate ROS that cause ferroptosis. The biological characteristics of ferroptosis are characterized by the aggregation of iron and ROS. When ferroptosis occurs, the cell membrane ruptures to form blisters, there is a lack of chromatin condensation in the cell nucleus, mitochondria decrease, the size of mitochondria becomes smaller, the density of the double membrane increases, the cristae of mitochondria decrease or disappear, and the outer membrane of mitochondria ruptures when observed under an electron microscope.

[0068] Ferroptosis is distinct from apoptosis and necroptosis in that it is mediated independently in the absence of the important effectors of apoptosis and necroptosis, BAX, BAK, caspases, mixed lineage kinase domain-like protein (MLKL), and receptor-interacting serine / threonine kinases (RIPK1 and RIPK3). Ferroptosis has an effect of preventing cancer in uncontrollable tumor cells, but it may be the cause underlying the onset of various diseases and may also be the hidden cause of many unconfirmed disease mechanisms. Although intensive research has been conducted in the field of ferroptosis, many more studies are needed to fully understand its mechanism and role in physiological and pathological conditions. So far, the mechanism of ferroptosis described by various researchers may be simply composed of four steps: (i) inactivation of the cysteine / glutathione antiporter system, (ii) depletion of glutathione and GPx4, (iii) excessive production of lipid ROS, and (iv) excessive intracellular iron accumulation.

[0069] Ferroptosis spreads paracrinally via signals that have not yet been clearly identified and may involve 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA), the toxic end products of lipid peroxidation, which are stable and can react with biological macromolecules and affect sites distant from the source of generation. Observation by transmission electron microscopy shows that a nucleus through which electrons can pass is a unique feature of ferroptosis and is known to be such. The morphological features induced in cells by ferroptosis, unlike apoptosis, mainly affect mitochondria. Dixon et al. observed mitochondrial shrinkage and dysfunction and suggested that ferroptosis is directly related to cellular energy and that mitochondrial dysfunction leads to changes and a deficiency of cellular energy and ultimately cell death. The typical profile of ferroptosis is regulated cell death by iron-dependent lipid peroxidation, which can be ameliorated by iron chelators and lipid antioxidants. The cause of lipid peroxidation is generally thought to be reactive oxygen species (ROS) that react with polyunsaturated fatty acids (PUFAs) in the membrane to induce lipid peroxidation. Although several ROS generation pathways have been proposed, the detailed mechanism of iron-induced ROS remains unclear.

[0070] As shown in Figure 1, ferroptosis plays an important role in various diseases of various organs. (Original Figure 1 from Li J. et al. Cell Death and Disease (2020) 11:88. Accessed on January 28, 2022, at https: / / doi.org / 10.1038 / s41419-020-2298-2). Ferroptosis is also involved in the vicious cycle of the aging process, as shown in Figure 2 (from the original Figure 1 published in Mazhar M, et al. Cell Death Discovery (2021) 7:149. Accessed on January 28, 2022, at https: / / doi.org / 10.1038 / s41420-021-00553-6).

[0071] It is widely recognized that chelating iron can improve / prevent iron deficiency. However, the chelating agents proposed in the literature are only deferoxamine (DESFERAL®), which is approved only for intravenous and subcutaneous administration, 2,2'-bipyridyl (2,2'-dipyridine), a highly toxic substance that shows effectiveness in cell culture, and ciclopirox olamine (LOPROX®), a chelating bactericide for topical use that is not approved for systemic use. All of these have safety issues, are not suitable for long-term use to prevent ongoing ferroptosis, and have never been evaluated at non-toxic doses for long-term use.

[0072] Some iron chelating agents, such as disodium EDTA, calcium disodium EDTA, and metaphosphates, are commonly used in food. These compounds are generally not absorbed into the body by oral administration, but they may be used for oral administration to chelate iron and prevent or improve iron deficiency.

[0073] Chelating agent: Chelation is a chemical bond with a metal in a complex in which the metal is part of a ring. The organic ligand is called a chelating agent or chelator, and the chelate is a metal complex. The more closed rings there are to the metal atom, the more stable the compound. The stability of the chelate is also related to the number of atoms in the chelate ring. H 2 O or NH 3Monodentate ligands with one coordinating atom, such as , are easily decomposed by other chemical processes, while multidentate chelating agents that provide multiple bonds to metal ions offer more stable complexes. Chlorophyll, a green plant pigment, is a chelate compound in which a central magnesium atom is bonded to four complex chelating agents (pyrrole rings). Heme is an iron chelate containing an iron(II) ion at the center of porphyrin. Chelating agents provide a wide range of metal ion sequestering agents for controlling metal ions in aqueous systems. Chelating agents form stable water-soluble complexes with polyvalent metal ions, inhibiting the normal reactivity of metal ions and preventing unwanted interactions. EDTA (ethylenediaminetetraacetic acid) is a good example of a common chelating agent with nitrogen atoms and short-chain carboxyl groups.

[0074] For the purposes of the present invention, those that can be safely ingested are suitable for formulating compositions that can be ingested by subjects in need of iron chelation. Examples of chelating agents for iron and calcium include diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), 1,3-propylenediaminetetraacetic acid (PDTA), ethylenediaminedisuccinic acid (EDDS), ethylene glycol tetraacetic acid (EGTA), and the like. Any suitable chelating agent known in the art that is biologically safe and can chelate iron, calcium, or other metals is suitable for the present invention.

[0075] The concentration of the chelating agent needs to be sufficient so that the chelating agent can pass through the cell membrane and access the iron inside the cell. Therefore, the concentration of the chelating agent is determined by the thickness of the cell membrane of the animal or human subject ingesting the formulation. For example, in an exemplary evaluation, rats were freely administered drinking water containing 26 ppm (0.0026%) of disodium EDTA, which corresponds to 260 ppm (0.026%) when adjusted for humans. In some embodiments, the transport enhancer can be present in the formulations of the present invention in an amount in the range of about 0.0001 wt% to about 15 wt%, typically in the range of about 0.001 wt% to about 1 wt%, more typically in the range of about 0.10 wt% to about 5 wt%.

[0076] Compounds useful as chelating agents herein include any compound that coordinates with or forms a complex with a divalent or polyvalent metal cation and thereby functions as a sequestering agent for such cations. Thus, the term "chelating agent" as used herein includes not only divalent and polyvalent ligands (commonly referred to as "chelating agents"), but also monovalent ligands that can coordinate with or form a complex with a metal cation.

[0077] Suitable biocompatible chelating agents useful in combination with the present invention include, but are not limited to, EDTA, cyclohexanediaminetetraacetic acid (CDTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylenephosphonic acid (ATPA), citric acid, pharmaceutically acceptable salts thereof, and monomeric polyacids such as combinations of any of the foregoing. Other exemplary chelating agents include phosphoric acid, such as pyrophosphoric acid, tripolyphosphoric acid, hexametaphosphoric acid, malic acid polymers, and the like.

[0078] EDTA and acceptable EDTA salts are particularly preferred, and representative acceptable EDTA salts are usually selected from EDTA diammonium, EDTA disodium, EDTA dipotassium, EDTA triammonium, EDTA trisodium, EDTA tripotassium, and calcium disodium EDTA.

[0079] EDTA is widely used as an agent for chelating metals in biological tissues and blood and has been proposed to be included in various formulations. For example, U.S. Patent No. 6,348,508 to Denick Jr., etc. EDTA functions as a metal ion sequestering agent that binds metal ions. In addition to its use as a chelating agent, EDTA is also widely used as a preservative in place of benzalkonium chloride, as described, for example, in U.S. Patent No. 6,211,238 to Castillo et al. U.S. Patent No. 6,265,444 to Bowman et al. discloses the use of EDTA as a preservative and stabilizer. However, since EDTA has low permeability through biological membranes, it is generally not topically applied in high-concentration formulations.

[0080] Among the chelating / sequestering substances that can be included in the composition, biocompatible chelating agents include, but are not limited to, EDTA, cyclohexanediaminetetraacetic acid (CDTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylenephosphonic acid (ATPA), citric acid, pharmaceutically acceptable salts thereof, and monomeric polyacids such as any combination of the foregoing.

[0081] Other exemplary chelating agents include phosphoric acids such as pyrophosphoric acid, tripolyphosphoric acid, and hexametaphosphoric acid. Other exemplary chelating agents include phosphoric acids such as pyrophosphoric acid, tripolyphosphoric acid, hexametaphosphoric acid, chelating antibiotics such as chloroquine and tetracycline, and nitrogen-containing chelating agents containing two or more chelating nitrogen atoms within an imino group or an aromatic ring (e.g., diimine, 2,2'-bipyridine, etc.). And polyamines such as cyclam (1,4,7,11-tetraazacyclotetradecane), N-(C 1 -C 30Alkyl)-substituted cyclams (e.g., hexadecylcyclam, tetramethylhexadecylcyclam), diethylenetriamine (DETA), spermine, diethylnorspermine (DENSPM), diethylhomospermine (DEHOP), deferoxamine (N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide, or N'-[5-(acetylhydroxyamino)pentyl]-N-[5-[3-(5-aminopentylhydroxycarbamoyl)propanoylamino]pentyl]-N-hydroxybutanediamide); also known as desferrioxamine B, desferrioxamine B, DFO-B, DFOA, DFB or desferal), deferiprone, pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), ethane-1,2-bis(N-1-amino-3-ethylbutyl-3-thiol).

[0082] Additional suitable biocompatible chelating agents that may be useful in the practice of the present disclosure include EDTA-4-aminoquinoline complexes such as ethyl ([2-(bisethoxycarbonylmethylamino)-ethyl]-{[2-(7-chloroquinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([2-(bisethoxycarbonylmethylamino)-propyl]-{[2-(7-chloroquinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([3-(bisethoxycarbonylmethylamino)-propyl]-{[2-(7-chloroquinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([4-(bisethoxycarbonylmethylamino)-butyl]-{[2-(7-chloroquinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetate, and the like. Ethyl ([2-(bis-ethoxymethylamino)-ethyl]-{[2-(7-chloroquinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([2-(bis-ethoxymethylamino)-propyl]-{[2-(7-chloroquinolin-4-ylamino)0)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([3-(bis-ethoxymethylamino)-propyl]-{[2-(7-chloroquinolin-4-ylamino)1)-ethylcarbamoyl]-methyl}-amino)-acetate, ethyl ([4-(bis-ethoxymethylamino)-butyl]-{[2-(7-chloroquinolin-4-ylamino)2)-ethylcarbamoyl]-methyl}-amino)-acetate as described in Solomon et al., Med. Chem. 2: 133-138, 2006. (7-chloroquinolin-4-ylamino)(7-chloroquinolin-4-ylamino) (7-chloroquinolin-4-ylamino)

[0083] Furthermore, natural chelating agents include, but are not limited to, citric acid, phytic acid, lactic acid, acetic acid, and their salts. Other natural chelating agents and weak chelating agents include, but are not limited to, curcumin (turmeric), ascorbic acid, succinic acid, and the like.

[0084] In some embodiments, the chelating agent is selected from the tetrasodium salt of iminodisuccinic acid (Baypure® CX100, LANXESS GMBH (formerly Bayer Chemicals) Leverkusen, DE) or the salt of polyaspartic acid (Baypure® DS100; LANXESS GMBH, Leverkusen, Delaware). In some embodiments, the chelating agent is the tetrasodium salt of L-glutamic acid N,N-diacetic acid (GLDA - Dissolvine®, AkzoNobel, Netherlands).

[0085] In some embodiments, the chelating agent incorporated into the formulation is a prochelating agent. A prochelating agent is a molecule that is converted into a chelating agent when exposed to appropriate chemical or physical conditions. For example, the BSIH (isonicotinic acid [2-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-benzylidene]-hydrazide) prochelating agent is converted by hydrogen peroxide into the SIH (salicylaldehyde isonicotinoyl hydrazone) iron chelating agent that inhibits the generation of hydroxyl radicals by an iron catalyst.

[0086] The inactivated metal ion sequestering agent may also be referred to herein as a "prochelating agent", but the sequestration of metal ions may involve sequestration and complexation processes that extend beyond the scope of chelation itself. The term "prochelator" is similar to the term "prodrug", where a prodrug is a therapeutically inactive agent until it is activated in the body, and a prochelator similarly cannot sequester metal ions until it is activated in the body.

[0087] Transport enhancer: The transport promoter is selected to promote the transport of the chelating agent through tissues in the body, the extracellular matrix, and / or cell membranes. The "effective amount" of the transport promoter refers to an amount and concentration in the formulation of the present invention that is sufficient to measurably increase the penetration of the chelating agent passing through one or more sites in the oral cavity or teeth of a subject compared to when the transport promoter is not included in the formulation.

[0088] The concentration of the transport promoter needs to be sufficient for the chelating agent to move through the cell membrane. Thus, the concentration or relative amount of the transport promoter is determined by the thickness of the cell membrane of the animal or human subject ingesting the formulation. In some embodiments, the concentration of MSM in the formulation ranges from about 0.0001% to 30% by weight, or from about 0.01% to about 0.10, 1, 5, 10, 20, 30% by weight, preferably from about 0.01% to 1.0% by weight.

[0089] The transport promoter is generally represented by formula (I).

Chemical formula

[0090] The phrases "having a formula" or "having a structure" are not intended to be limiting and are used in the same way as the term "comprising" is commonly used. In the above structure, the term "alkyl" refers to a straight-chain, branched, or cyclic saturated hydrocarbon group containing 1 to 6 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, cyclopentyl, cyclohexyl, etc. Unless otherwise indicated, the term "alkyl" includes unsubstituted alkyl and substituted alkyl, and examples of substituents include halo, hydroxyl, sulfhydryl, alkoxy, acyl, etc. The term "alkoxy" means an alkyl group bonded through a single terminal ether bond. That is, an "alkoxy" group is represented as -O-alkyl where alkyl is as defined above. The term "aryl" refers to an aromatic substituent containing a single aromatic ring or a plurality of aromatic rings fused, directly bonded, or indirectly bonded to each other (where different aromatic rings are bonded to a common group such as a methylene or ethylene moiety). Preferred aryl groups contain 5 to 14 carbon atoms. Exemplary aryl groups include one aromatic ring or two fused or bonded aromatic rings (e.g., phenyl, naphthyl, biphenyl, diphenyl ether, diphenylamine, benzophenone, etc.). "Aryl" includes unsubstituted aryl and substituted aryl, and the substituents are as described above for an optionally substituted "alkyl" group. The term "aralkyl" refers to an alkyl group having an aryl substituent, where "aryl" and "alkyl" are as defined above. Preferred aralkyl groups contain 6 to 14 carbon atoms, and particularly preferred aralkyl groups contain 6 to 8 carbon atoms. Examples of aralkyl groups include, but are not limited to, benzyl, 2-phenyl-ethyl, 3-phenyl-propyl, 4-phenyl-butyl, 5-phenyl-pentyl, 4-phenylcyclohexyl, 4-benzylcyclohexyl, 4-phenylcyclohexylmethyl, 4-benzylcyclohexylmethyl, etc.The term "acyl" refers to a substituent having the formula -(CO)-alkyl, -(CO)-aryl, or -(CO)-aralkyl, where "alkyl", "aryl", and "aralkyl" are as defined above. The terms "heteroalkyl" and "heteroaralkyl" are used to refer to alkyl and aralkyl groups containing heteroatoms, respectively, i.e., alkyl and aralkyl groups in which one or more carbon atoms are replaced by atoms other than carbon, such as nitrogen, oxygen, sulfur, phosphorus, or silicon, typically nitrogen, oxygen, or sulfur.

[0091] This formulation also contains an effective amount of a transport promoter that promotes the penetration of the formulation components through cell membranes, tissues, and the extracellular matrix. The "effective amount" of the transport promoter, as described above, represents a concentration sufficient to measurably increase the penetration of one or more of the formulation components through the membrane, tissue, and extracellular matrix. Suitable transport promoters include, for example, methylsulfonylmethane (MSM, also called methyl sulfone), a combination of MSM and dimethyl sulfoxide (DMSO), or a combination of MSM and, in less preferred embodiments, DMSO, with MSM being particularly preferred. DMSO is a transport promoter but is essentially a solvent and is not particularly suitable for the formulations of the present invention. DMSO functions as a very strong solvent and thus functions as a carrier for its solutes. In contrast, MSM functions in a completely different way by forming hydrogen bonds with the selected molecule and changing the charge characteristics of the target molecule, enabling the target molecule to pass through charged barriers such as biological membranes.

[0092] There are differences in the chemical structures of MSM and DMSO. Methylsulfonylmethane (MSM) is an organosulfur compound represented by the chemical formula (CH 3 ) 2 SO 2 . DMSO 2 , also known by several other names such as methyl sulfone, dimethyl sulfone. This colorless solid is characterized by a sulfonyl functional group and is considered to be chemically relatively inert. The structure of MSM is as follows. [Chemical formula] On the one hand, dimethyl sulfoxide (DMSO) is an organosulfur compound represented by the chemical formula (CH 3 ) 2 SO. This colorless liquid is a widely used polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible with not only water but also a wide range of organic solvents. The structure of DMSO is as follows. [Chemical formula]

[0093] MSM is an odorless and highly water-soluble (34% w / v at 79°F) white crystalline compound with a melting point of 108 - 110°C and a molecular weight of 94.1 g / mol. MSM not only enhances cell membrane permeability but also functions as a "transport facilitator" (TFA) that aids in the transport of one or more formulation components to oral tissues, thus functioning as a multifunctional agent in this specification. Furthermore, MSM itself has medicinal effects and also functions as an anti-inflammatory and analgesic agent. MSM also has the function of improving the oxidative metabolism of biological tissues and is a source of organic sulfur that helps reduce scars. MSM is further unique and beneficial in having solubilization properties in that it dissolves in water as described above, but exhibits both hydrophilic and hydrophobic properties due to the presence of polar S=O groups and nonpolar methyl groups. The molecular structure of MSM allows for the formation of hydrogen bonds with other molecules, i.e., bonds between the oxygen atoms of each S=O group and the hydrogen atoms of other molecules, as well as van der Waals bonds, i.e., bonds between the methyl groups and nonpolar (such as hydrocarbon) segments of other molecules.

[0094] Formulation To formulate the compositions of the present invention, various means can be used. For techniques of formulation and administration, see "Remington: The Science and Practice of Pharmacy", 23rd Edition, Editor-in-Chief Adeboye Adejare, Academic Press, 23rd Edition (November 13, 2020). When administered to humans or animals, the formulations need to meet the standards of sterility, pyrogenicity, general safety, and purity equivalent to those required by the FDA. Administration of pharmaceutical formulations can be carried out in various ways, some of which are described herein.

[0095] Other possible additives that can be incorporated into at least partially aqueous formulations include, but are not limited to, thickeners, tonicity agents, buffers, and preservatives, provided that such excipients interact in a way that does not adversely affect any of the other components of the formulation. It should also be noted that preservatives are not necessarily generally required, considering the fact that the selected chelating agent itself serves as a preservative.

[0096] Chelating agents and penetration enhancers are dissolved in solvents selected from the non-limiting list of available solvents such as water, ethanol, acetone, DMSO, isopropanol, glycerol, propylene glycol, polyethylene glycol, propylene carbonate, and ethyl acetate.

[0097] In some embodiments, the formulation further comprises an emulsifier, and the emulsifier is gum arabic, modified starch, pectin, xanthan gum, ghatti gum, tragacanth gum, fenugreek gum, mesquite gum, monoglycerides and diglycerides of long-chain fatty acids, sucrose monoesters, sorbitan esters, polyethoxylated glycerol, stearic acid, palmitic acid, monoglycerides, diglycerides, propylene glycol esters, lecithin, emulsifying monoglycerides and diglycerides, propylene glycol monoesters, polyglycerol esters, diacetyl tartaric acid esters of monoglycerides and diglycerides, citric acid esters of monoglycerides, stearoyl-2-lactate, polysorbate, succinylated monoglycerides, acetylated monoglycerides, ethoxylated monoglycerides, quillaia, whey protein isolate, casein, soy protein, vegetable protein, pullulan, sodium alginate, guar gum, locust bean gum, tragacanth gum, tamarind gum, carrageenan, fucellaran, gellan gum, psyllium, curdlan, konjac mannan, agar, cellulose derivatives, or combinations thereof.

[0098] In some embodiments, the beverage formulation further comprises a flavor selected from the group consisting of vanilla, vanillin, ethyl vanillin, orange oil, peppermint oil, strawberry, raspberry, and mixtures thereof. The flavor may be other synthetic flavors, natural flavors, or combinations thereof.

[0099] In some embodiments, the formulation is aceclofenac, aspirin, celecoxib, clonixin, dexibuprofen, dexketoprofen, diclofenac, diflunisal, droxicam, etodolac, etoricoxib, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen, indomethacin, isoxicam, ketoprofen, ketorolac, licofelone, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimesulide, oxaprozin, parecoxib, phenylbutazone, piroxicam, rofecoxib, salsalate, sulindac, tenoxicam, tolfenamic acid, tolmetin, or valdecoxib.

[0100] In some embodiments, the ready-to-drink beverage comprises an infusion of tea leaves, coffee beans, or cocoa powder.

[0100] Any suitable isotonic agent and buffer commonly used in oral formulations can be used, provided that the pH of the formulation is maintained in the range of about 6.0 to about 9.0, preferably in the range of about 7.0 to about 7.4.

[0101] In some embodiments, the composition is a ready-to-drink beverage selected from the group consisting of non-carbonated beverages, carbonated beverages, colas, root beers, fruit-flavored beverages, citrus-flavored beverages, fruit juices, fruit-containing beverages, vegetable juices, vegetable-containing beverages, tea, coffee, dairy beverages, protein-containing beverages, shakes, sports drinks, energy drinks, and flavored water.

[0102] In a preferred embodiment, the pharmaceutical preparation is administered in the form of an orally ingestible liquid. However, the preparation may be available in any form that can be easily reconstituted into a liquid concentrate, soluble solid, effervescent tablet, or any form that can be easily reconstituted into a potable liquid. The concentrate can be made into a unit dosage form suitable for a single administration of an exact dosage. Suitable pharmaceutical preparations and dosage forms are known to those skilled in the art of pharmaceutical preparations and can be prepared using conventional methods described in relevant texts and literature such as "Remington: The Science and Practice of Pharmacy" previously mentioned herein, for example.

[0103] The chelating agent can be administered, as necessary, in the form of salts, esters, crystal forms, hydrates, etc., as long as it is pharmaceutically acceptable. Salts, esters, etc. are known to those skilled in the field of organic synthetic chemistry and can be prepared using standard procedures described, for example, in March's Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 6th Edition, edited by Smith MB and March J (Wiley-Interscience, 2007).

[0104] The amount of chelating agent administered depends on many factors, varies from subject to subject, and differs depending on the specific chelating agent, the specific disorder or condition being treated, the severity of the symptoms, the age, weight, general condition of the subject, and the judgment of the prescribing physician. The term "dosage form" refers to any form of a pharmaceutical composition containing an amount of chelating agent and transport promoter sufficient to achieve a therapeutic effect in a single or multiple administrations. The dosing frequency that most effectively results in the most effective outcome without over-dosing varies depending on characteristics including both the pharmacological properties of the specific active agent and physical properties such as hydrophilicity.

[0105] The complex may also contain conventional additives such as solvents, flavoring agents, antioxidants, fragrances, colorants, stabilizers, surfactants, etc. Other agents such as antibacterial agents can also be added to prevent spoilage during storage, i.e., to inhibit the growth of microorganisms such as yeast and mold. Suitable antibacterial agents are typically selected from methyl and propyl esters of p-hydroxybenzoic acid (i.e., methylparaben and propylparaben), sodium benzoate, sorbic acid, imidurea, and combinations thereof.

[0106] Pharmaceutical dosage forms suitable for ingestion include aqueous solutions containing the active ingredient. In any case, the final dosage form needs to be a stable liquid under the conditions of manufacture and storage. The liquid carrier or medium can be a solvent or liquid dispersion medium containing, for example, water, ethanol, polyols (such as glycerol, propylene glycol, liquid polyethylene glycol, etc.), and suitable mixtures thereof. In many cases, it may be desirable to include sugars, buffering agents, isotonic agents such as sodium chloride.

[0107] The treatment plan varies depending on several factors that can be easily determined, such as the severity of the symptoms and the responsiveness to the symptoms to be treated, but usually treatment is carried out more than once a day. Treatment generally consists of the ingestion of one or more types of drinking water containing an iron chelating agent and a penetration enhancer. The treatment period continues from one day or several days to several months, or until the iron ions in the body are kept at a desirable level or the state of iron deficiency is significantly reduced.

[0108] The composition of the present invention may further contain additional agents or excipients depending on the indication. In one aspect of the embodiment, the pharmaceutical composition further contains a therapeutically effective amount of at least one antibacterial or antifungal agent. More specifically, the antibacterial agent is an antibiotic.

Examples

[0109] The following examples are presented to provide those skilled in the art with a complete invention and an explanation of how to create and use embodiments in accordance with the present invention, and are not intended to limit the scope that the inventors regard as their discoveries. Although efforts are made to ensure the accuracy of the numerical values (amounts, temperatures, etc.) used, some experimental errors and deviations need to be considered. Unless otherwise stated, parts are by weight, molecular weight is weight-average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

[0110] To evaluate the ability of the formulations according to the present invention, rats were allowed free access to drinking water containing 26 ppm (0.0026) disodium EDTA. Adjusted for human use, this corresponds to 260 ppm (0.026%) for free intake. Since humans cannot freely ingest it, in order to obtain equivalent results, the chelating agent needs to be up-regulated to 500 - 1000 ppm or up to 0.5%. In the case of animals that can freely drink water, equivalent results can be obtained at 10 ppm or more depending on the body weight and size of the animal.

[0111] Experiments were conducted using rats to examine the usefulness of these concentrations in drinking water.

[0112] Example 1: Inhibition of ferroptosis by iron chelation in a TNBS-induced colitis model. TNBS colitis is recognized as an animal model of chemically induced colitis used to test reagents that affect Crohn's disease. The hapten reagent 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis is used in preclinical trials of various chemical substances and natural compounds from the viewpoints of anti-inflammatory and antioxidant effects. Ferroptosis has been shown to be involved in inflammatory bowel disease (IBD) consisting of ulcerative colitis (UC) and Crohn's disease (CD). Ferroptosis is a recently recognized form of regulated cell death (RCD) and is identified as iron-dependent and caspase-independent non-apoptotic cell death.

[0113] In the TNBS model, H&E (hematoxylin and eosin) staining of the intestinal mucosal structure was observed on the third day after colitis induction. This is a widely accepted model that also covers ferroptosis in diabetes. Iron chelation has been shown to alleviate TNBS-induced colitis through the inhibition of ferroptosis (Xu J, et al., Biochemical and Biophysical Research Communications 573(2021)48-54).

[0114] Figure 3 shows the H&E-stained intestinal mucosa 3 days after inducing colitis with TNBS. Control: The mucosal structure is complete and there is no cell infiltration in the submucosa. TNBS: Colonic mucosal structure damage, submucosal cell infiltration. TNBS + ME (ME: MSM + EDTA): The colonic structure shows slight changes and a small number of cell infiltrations are observed in the submucosa.

[0115] Example 2: Inhibition of ferroptosis by iron chelation in the DSS-induced colitis model Dextran sodium sulfate (DSS) is a sulfated polysaccharide with various molecular weights. When DSS is administered, the mucosal barrier function decreases due to its toxicity to colonic epithelial cells, causing a pathological condition similar to human ulcerative colitis. Iron chelation has been shown to alleviate DSS-induced colitis through the inhibition of ferroptosis (Chen Y, et al., Immunology Letters 225(2020)9-15).

[0116] Figure 4 shows the H&E-stained intestinal mucosa 3 days after inducing colitis with DSS. Control: The mucosal structure is complete and there is no cell infiltration in the submucosa. DSS: Colonic mucosal ulcer (arrow), decrease and structural damage of goblet cells, submucosal edema, cell infiltration. DSS + ME (MSM + EDTA): The colonic structure shows slight changes and a small number of cells are infiltrating in the submucosa.

[0117] Example 3: Immunohistochemistry (IHC) of iron chelation and inhibition of ferroptosis in the DSS-induced colitis model. One week after colitis induction, immunohistochemistry (IHC) using ALDH1, protein-HNE, protein-acrolein, and iNOS staining was performed in the DFSS colitis model. This is a widely accepted model that also covers ferroptosis in diabetes (Chen 2020). 4-HNE (4-hydroxynonenal) and iNOS are recognized as markers of ferroptosis (Yan Hf, et al., Signal Transduction and Targeted Therapy (2021) 6:49)

[0118] Figure 5 shows IHC staining with ALDH1, protein-HNE, protein-acrolein, and protein-MDA in the DSS model of rat colitis, indicating the reduction of microscopic colonic damage by MSM+EDTA (ME) treatment. The upper panel is anti-protein-HNE staining, and the lower panel is anti-protein-HNE using DAPI. Normal control. B: DSS-induced colitis. C: DSS-induced colitis treated with ME.

[0119] Example 4: Lens Opacity in Diabetic Rats Opacity (cataract) of the lens and the ability to reduce activity levels were observed in rats induced with diabetes. The results are shown in the table below. CTRL is the control group of non-diabetic rats administered normal water, DT is the group of diabetic rats administered drinking water injected with the test MSM / EDTA, and DC is the group of diabetic control rats administered normal (other than MSM / EDTA) drinking water. The cataract scale is 0-4 (4 being the most severe).

Table 1

[0120] The statistical significance of the test was established by determining the ANOVA p-value. When the p-value is below the threshold of p<0.05, the results of the test are statistically significant.

Table 2

[0121] The activity levels of diabetic rats administered with MSM and iron chelating agents were observed, and the results are shown below.

Table 3

[0122] Example 5: Effect of iron chelation on chronic inflammation using IL-6 as a marker in rats Normal water or water added with MSM and chelating agent was freely administered to rats, and chronic inflammation was examined using IL-6 as a marker. The MSM / EDTA concentration in the water was 26 ppm for EDTA and 54 ppm for MSM. NR: Normal rats administered with normal water, NR+ME: Normal rats + MSM / EDTA drinking water. DR: Diabetic rats administered with normal water, DR+ME: Diabetic rats + MSM / EDTA drinking water.

[0123] Figure 6 shows chronic inflammation using IL-6 as a marker in rats that freely ingested normal water or water added with MSM and chelating agent. The MSM / EDTA concentration in the water was 26 ppm for EDTA and 54 ppm for MSM. NR: Normal rats administered with normal water, NR+ME: Normal rats + MSM / EDTA drinking water. DR: Diabetic rats administered with normal water, DR+ME: Diabetic rats + MSM / EDTA drinking water. After administration of MSM and iron chelating agent, the IL-6 level in diabetic rats decreased to one-fourth.

[0124] Example 6: Health of pancreatic endocrine islets after administration of iron chelating agent in diabetic rats Figure 7A shows low magnification (100x) microscopic photographs of H&E stained 4μm sections of formalin-fixed paraffin-embedded pancreas, showing pancreatic lobules. Sections of the pancreas of normal rats (A) show normal numbers and sizes of endocrine islets of Langerhans and normal exocrine acinar tissue. (B) Normal islets and acinar tissue are also seen in normal rats orally administered M+E. (C) shows that the number and size of pancreatic endocrine islets of Langerhans in diabetic rats are significantly decreased. Most of the islets were small, shrunken and inconspicuous. (D) Oral administration of M+E to diabetic rats significantly improved the number and size of endocrine islets of Langerhans, and no shrinkage of acinar tissue was observed. Figure 7B shows high magnification (400x) microscopic photographs of pancreatic endocrine islets, showing H&E stained 4μm sections of formalin-fixed paraffin-embedded pancreas. (A) In the endocrine islets of the pancreas of normal rats, cells scattered within lightly stained exocrine acini, spherical cell clusters without ducts, and acini are seen. (B). No histologically and morphologically significant changes were observed in the endocrine islets of pancreatic sections of normal rats orally administered. (C) In the endocrine islets of the pancreas of diabetic rats, the islets of Langerhans are shrunk and become small and inconspicuous (sclerosis of the islets, decrease in the cytoplasm of cells), and the presence of interstitial pancreatitis is seen, as evident from leukocyte infiltration within the islets. (D). In the endocrine islets of diabetic rats orally administered, it is shown that the islets of Langerhans are slightly shrunk and leukocyte infiltration is minimal.

[0125] All publications and patent applications cited herein are hereby incorporated by reference into this specification, as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

[0126] The foregoing invention has been described in detail by way of illustration and example for purposes of clarity of understanding, but it will be apparent to those skilled in the art that certain changes and modifications can be made without departing from the spirit or scope of the appended claims in light of the teachings of the present invention.

Claims

1. A formulation comprising the following components: A chelating agent or a salt thereof, wherein the chelating agent is suitable for long-term ingestion; Methylsulfonylmethane (MSM), which is a penetration enhancer One or more inert excipients, and A liquid medium or carrier. The chelating agent and the penetration enhancer are present in a proportion effective to maintain the homeostasis of iron levels in the body when ingested at normal doses, The proportion of the chelating agent is from about 0.0001% to 15%, and the proportion of the penetration enhancer in the composition is from about 0.0001% to 30% by weight, respectively.

2. The formulation according to claim 1, wherein the homeostasis of iron levels is sufficient to inhibit the replication of RNA viruses in a human or animal subject.

3. The formulation according to claim 2, wherein the RNA virus is selected from HIV-1, SARS-CoV-2, MERS, SARS, HTLV-I, and HTLV-II.

4. The formulation according to claim 1, wherein the homeostasis of iron levels regulates ferroptosis in the subject.

5. The formulation according to claim 4, wherein ferroptosis is associated with a pathological disease or condition in the subject.

6. The formulation according to claim 5, wherein the pathological disease or condition is associated with an organ selected from the heart, central nervous system, liver, gastrointestinal organs, lungs, kidneys, and pancreas.

7. The formulation according to claim 4, wherein ferroptosis is associated with aging in the subject.

8. The formulation according to claim 4, wherein ferroptosis is associated with inflammation in the subject

9. The formulation according to claim 4, wherein ferroptosis is reduced by inhibiting the accumulation of iron-dependent lipid reactive oxygen species (ROS).

10. The formulation according to claim 1, wherein the homeostasis of iron levels is maintained by chelating iron bound to a heme-containing protein selected from hemoglobin, myoglobin, and neuroglobin.

11. The formulation according to claim 1, wherein the homeostasis of iron levels is related to the regulation of ferroptosis in human cancer cells.

12. The formulation according to claim 1, wherein the homeostasis of iron levels is related to the regulation of ferroptosis in a disease selected from cancer, neurodegenerative diseases, and I / R injury-related diseases.

13. The formulation according to claim 1, wherein the chelating agent is selected from ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), cyclohexanediaminetetraacetic acid (CDTA), hydroxyethylethylenediaminetriacetic acid (HEDTA), diethylenetriaminepentaacetic acid (DTPA), dimercaptopropanesulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylenephosphonic acid (ArPA), citric acid, acetic acid and their acceptable salts, and any combination thereof.

14. The formulation according to claim 12, wherein the EDTA salt is selected from EDTA diammonium, EDTA disodium, EDTA dipotassium, EDTA triammonium, EDTA trisodium, EDTA tripotassium, EDTA tetrasodium, EDTA tetrapotassium, calcium disodium EDTA, and combinations thereof.

15. The formulation according to claim 1, wherein the chelating agent is selected from phosphate, pyrophosphate, tripolyphosphate, and hexametaphosphate.

16. The formulation according to claim 1, wherein the chelating agent is a nitrogen-containing chelating agent containing two or more chelating nitrogen atoms within an imino group or an aromatic ring, a diimine, or 2,2'-bipyridine.

17. The formulation according to claim 1, wherein the chelating agent is cyclam (1,4,7,11-tetraazacyclotetradecane), N-(C 1 -C 30 alkyl)-substituted cyclam (e.g., hexadecylcyclam, tetramethylhexadecylcyclam), diethylenetriamine (DETA), spermine, diethylnorspermine (DENSPM), diethylhomospermine (DEHOP), deferoxamine (N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide, or N'-[5-(acetylhydroxyamino)pentyl]-N-[5-[3-(5-aminopentylhydroxycarbamoyl)propanoylamino]pentyl]-N-hydroxybutanediamide), desferrioxamine B, desferrioxamine B, DFO-B, DFOA, DFB, desferal, deferiprone, a polyamine selected from the group consisting of. Pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), ethane-1,2-bis(N-1-amino-3-ethylbutyl-3-thiol).

18. The formulation according to claim 1, wherein the chelating agent is ethyl ([2-(bis(ethoxycarbonylmethyl)amino)ethyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([2-(bis(ethoxycarbonylmethyl)amino)propyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([3-(bis(ethoxycarbonylmethyl)amino)propyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([4-(bis(ethoxycarbonylmethyl)amino)butyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([2-(bis(ethoxymethyl)amino)ethyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, which are EDTA-4-aminoquinoline conjugates selected from ethyl ([2-(bis(ethoxymethyl)amino)propyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([3-(bis(ethoxymethyl)amino)propyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate, ethyl ([4-(bis(ethoxymethyl)amino)butyl]-{[2-(7-chloroquinolin-4-ylamino)ethylcarbamoyl]methyl}amino)acetate. (7-chloroquinolin-4-ylamino)(7-chloroquinolin-4-ylamino)(7-chloroquinolin-4-ylamino)

19. The formulation according to claim 1, wherein the chelating agent is the tetrasodium salt of iminodisuccinic acid.

20. The formulation according to claim 1, wherein the chelating agent is polyaspartic acid or a salt thereof.

21. The formulation according to claim 1, wherein the chelating agent is the tetrasodium salt of L-glutamic acid N,N-diacetic acid.

22. The preparation according to claim 1, wherein the chelating agent is a natural chelating agent selected from citric acid, phytic acid, lactic acid, acetic acid, and their salts, and curcumin.

23. The preparation according to claim 1, further comprising an antibiotic.

24. The preparation according to claim 1, wherein the liquid carrier is selected from water, ethanol, acetone, DMSO, isopropanol, glycerol, propylene glycol, polyethylene glycol, propylene carbonate, and ethyl acetate.

25. The preparation according to claim 1, wherein the preparation further comprises an emulsifier, and the emulsifier is gum arabic, modified starch, pectin, xanthan gum, ghatti gum, tragacanth gum, fenugreek gum, mesquite gum, monoglycerides and diglycerides of long-chain fatty acids, sucrose monoesters, sorbitan esters, polyethoxylated glycerol, stearic acid, palmitic acid, monoglycerides, diglycerides, propylene glycol esters, lecithin, emulsifying monoglycerides and diglycerides, propylene glycol monoesters, polyglycerol esters, diacetyl tartaric acid esters of monoglycerides and diglycerides, citric acid esters of monoglycerides, stearoyl-2-lactate, polysorbate, succinylated monoglycerides, acetylated monoglycerides, ethoxylated monoglycerides, quillaia, whey protein isolate, casein, soy protein, vegetable protein, pullulan, sodium alginate, guar gum, locust bean gum, tragacanth gum, tamarind gum, carrageenan, fucellaran, gellan gum, psyllium, curdlan, konjac mannan, agar, cellulose derivatives, or combinations thereof.

26. The formulation according to claim 1, wherein the formulation further comprises an anti-inflammatory agent which is a non-steroidal anti-inflammatory (NSAID) drug selected from the group consisting of aceclofenac, aspirin, celecoxib, clonixin, dexibuprofen, dexketoprofen, diclofenac, diflunisal, droxicam, etodolac, etoricoxib, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen, indomethacin, isoxicam, ketoprofen, ketorolac, licofelone, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimesulide, oxaprozin, parecoxib, phenylbutazone, piroxicam. Rofecoxib, salsalate, sulindac, tenoxicam, tolfenamic acid, tolmetin, or valdecoxib.

27. The formulation according to claim 1, wherein the ready-to-drink formulation further comprises a fragrance selected from the group consisting of vanilla, vanillin, ethyl vanillin, orange oil, peppermint oil, strawberry, raspberry, synthetic fragrances and natural fragrances, and mixtures thereof.

28. The formulation according to claim 1, wherein the concentration of the chelating agent is 0.002% to 0.1% w / v.

29. The formulation according to claim 1, wherein the concentration of MSM is 0.005% to 3% w / v.

30. The formulation according to claim 1, wherein the formulation is in the form of a ready-to-drink beverage.

31. The formulation according to claim 30, wherein the ready-to-drink beverage comprises an extract of tea leaves, coffee beans, or cocoa powder.

32. The formulation according to claim 30, wherein the ready-to-drink beverage has a pH of 7.0 to 8.

0. And 7.

4.

33. The formulation according to claim 30, wherein the ready-to-drink beverage is selected from the group consisting of non-carbonated beverages, carbonated beverages, cola, root beer, fruit-flavored beverages, citrus-flavored beverages, fruit juices, fruit-containing beverages, vegetable juices, vegetable-containing beverages, tea, coffee, dairy beverages, protein-containing beverages, shakes, sports drinks, energy drinks, and flavored water.

34. The formulation according to claim 30, wherein the formulation may be easily reconstituted into a liquid concentrate, a soluble solid, a foaming tablet, a pill, or any form that may be easily reconstituted into a drinkable beverage.

35. The formulation according to claim 30, further comprising an isotonic agent selected from saccharides, buffers, and sodium chloride.

36. The formulation according to claim 1, which is in a liquid form administrable by a route selected from the group consisting of oral, intranasal, inhalation, intravenous, intramuscular, transdermal, topical, rectal, vaginal, buccal, injection, sublingual, or combinations thereof.

37. The formulation according to claim 36, wherein the formulation is a liquid, a soluble solid, a foaming tablet, a pill, or a concentrate in a form that may be easily reconstituted.

38. The formulation according to claim 36, wherein the formulation is a rehydration drip solution further comprising electrolytes.

39. A method of inhibiting the replication of an RNA virus in a subject, comprising administering the formulation according to any one of claims 1 to 38 at a frequency and for a period sufficient to reduce the iron ion concentration in the body to below the amount necessary to reduce hyperferritinemia.

40. The method according to claim 39, wherein the RNA virus is selected from HIV-1, SARS-CoV-2, MERS, SARS, HTLV-I, and HTLV-II.

41. A method of treating or alleviating iron deficiency in a subject, comprising providing the formulated beverage according to any one of claims 1 to 38 at a frequency and for a period sufficient to reduce the iron ion concentration in the body to below the amount necessary for pathogenic-level iron deficiency.

42. The method according to claim 41, wherein ferroptosis is associated with a pathological disease or condition in the subject.

43. The method according to claim 42, wherein the pathological disease or condition is associated with an organ of a human or animal subject selected from the heart, central nervous system, liver, gastrointestinal organs, lungs, kidneys, and pancreas.

44. The method according to claim 41, wherein ferroptosis is associated with aging in the subject.

45. The method according to claim 41, wherein ferroptosis is associated with inflammation in the subject.

46. The method according to claim 41, wherein the reduction of ferroptosis is measured by inhibition of iron-dependent lipid reactive oxygen species (ROS) accumulation.

47. The method according to claim 41, wherein ferroptosis is related to human cancer cells.

48. The method according to claim 41, wherein ferroptosis is related to a disease selected from the group consisting of cancer, neurodegenerative diseases, and I / R injury-related diseases.

49. A method of chelating intracellular iron in a subject, the method comprising providing the formulated beverage according to any one of claims 1 to 38 at a frequency and for a period sufficient to reduce iron ion levels associated with a heme-containing protein selected from hemoglobin, myoglobin, and neuroglobin.

50. The method according to claim 49, wherein the heme-containing protein is present in cells selected from red blood cells, muscle cells, or nerve cells.

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