WO2023147169A2 - Novel liquid formulations for iron chelation - Google Patents

Abstract

Iron-chelating formulations, comprising a chelator (such as EDTA and its salts) and methylsulfonylmethane (MSM), are provided. The formulation can be a ready-to-drink beverage, a hydration drip, or any liquid that can be systemically administered to a subject. Formulations and methods for treatment of RNA viruses including SARS-CoV-2 using the disclosed composition is provided. Formulations and methods for inhibition of ferroptosis and diseases and conditions affected by ferroptosis is provided. Among diseases and conditions affected by ferroptosis are cancer, aging, inflammation, and the like.

Description

NOVEL LIQUID FORMULATIONS FOR IRON CHELATION

INVENTORS:

Rajiv BHUSHAN and Amit GOSWAMY

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application Serial No. 63/305,099 titled " NOVEL LIQUID FORMULATIONS FOR IRON CHELATION," filed 31 January 2022, the contents of which are incorporated herein in their entirety by reference.

TECHNICAL FIELD OF THE INVENTION

[0002] This invention relates generally to the field of agents to treat ferroptotic cell death in cases of cancer, aging, viral infections and other ferroptosis-associated pathologic conditions. More particularly, it relates to consumable formulations containing a chelator and transport enhancers suitable for chelating intracellular iron. In particular, the invention relates to ready- to-drink beverage formulations containing a transport enhancer and a chelating agent. In one exemplary embodiment, it relates to drinkable liquid compositions which contain MSM as a transport enhancer and chelators generally acceptable as safe ("GRAS").

BACKGROUND OF THE INVENTION

[0003] Iron (Fe) plays a major role in human disease conditions. Certain viruses (RNA viruses) capable of causing lethal infections in humans are dependent on iron for their replication within the human body. For example, treatment with iron chelator deferiprone has been shown to prolong the survival of acquired immunodeficiency syndrome (AIDS) patients. Iron overload has been shown to cause ferroptosis which is part of multiple human diseases. Ferroptosis also has been implicated in debilitating aspects of the aging process.

Thus, limiting iron represents a promising adjuvant strategy in treating viral infection through oral uptake or venous injection of iron chelators, or through the manipulation of the key iron regulators.

[0004] Coronaviruses are a family of enveloped, single- stranded RNA viruses. In recent decades, two highly pathogenic strains of coronavirus were identified in humans: severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV). These viruses were found to cause severe, and sometimes fatal, respiratory illness. In December 2019, a new strain of coronavirus (SARS-CoV-2) causes Coronavirus disease 2019, or Covid-19, which was declared a pandemic by the WHO on 11 Mar. 2020. Common signs of Covid- 19 infection include respiratory symptoms, fever, cough, shortness of breath, and breathing difficulties. In more severe cases, the infection can cause pneumonia, severe acute respiratory syndrome, kidney failure and death. As of January 2022, over 373 million infections and 5.66 million deaths have been reported worldwide.

[0005] Expedited mass vaccination and mask- wearing protocols instituted worldwide may have reduced the severity of the disease in many patients, but the virus has demonstrated the ability to regularly mutate into newer variants that are highly lethal (e.g., delta), highly infectious (e.g., omicron) or have the ability to circumvent immunity generated by vaccines or infections with prior variants. However, no specific therapies presently exist, leaving the patients to depend on general and supportive therapies, including oxygen supply and broadspectrum antiviral medications. Remdesivir, a novel nucleotide analog prodrug in development for treating Ebola virus and Middle East Respiratory Syndrome (MERS) diseases, has been reported to relieve the pneumonia symptoms of CO VID- 19 infection. Given the global nature of the pandemic, there is an urgent need for medications and methods to inhibit or block replication of the SARS-CoV-2 virus and for such medications and methods to be widely and easily available to large sections of the global population who are all susceptible to the SARS-CoV-2 virus and its variants.

[0006] It has been demonstrated that iron (Fe) plays two major roles in Covid-19 disease and the severity of its symptoms. First, all coronaviruses (including SARS-Cov2) depend upon iron for their 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 developed to be highly dependent on iron for their replication and under low iron conditions, the replication slows down (Menshawey R, et al. Egyptian Journal of Medical Human Genetics (2020) 21:75). Increasing intracellular iron efflux via increasing iron exporter ferroportin expression also exhibits antiviral effect on human immunodeficiency virus (HIV). (Liu 2020). Second, Covid- 19 patients often suffer from iron overload, resulting in ferroptosis, which in turn leads to damage to 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] There is a role for hyper- ferritinemia and altered iron homeostasis in CO VID- 19 pathogenesis. Hyper-ferritinemia has been described as a cardinal feature that predicted with high significance the increased mortality risk from Covid- 19. (Mehta P, et al., Lancet 2020;395 (March (10229)): 1033-4). These studies demonstrated that serum ferritin levels in CO VID- 19 non-survivors exceeded the levels in the survivors by two-fold.

[0008] Ferroptosis is a type of regulated necrosis that is triggered by a combination of iron toxicity, lipid peroxidation, and plasma membrane damage. It is a programmed cell death pathway dependent upon intracellular iron, but not other metals. Non-apoptotic forms of cell death may facilitate the selective elimination of some tumor cells or be activated in specific pathological states. The oncogenic RAS-selective lethal small molecule erastin was found to trigger a unique iron-dependent form of non-apoptotic cell death. (Dixon SJ, et al. Cell. 2012 May 25; 149(5): 1060-1072). In 2012, Dixon coined the term ferroptosis, an iron dependent, non-apoptotic mode of cell death characterized by the accumulation of lipid reactive oxygen species (ROS). It is mainly caused by an increased redox imbalance but is morphologically, biochemically and genetically distinct from other known cell death patterns such as apoptosis, necrosis and autophagy.

[0009] Ferroptotic cells and their spilled contents shape innate and adaptive immunity in health and disease. Excessive or deficient ferroptotic cell death is implicated in a growing list of physiological and pathophysiological processes, coupled with a dysregulated immune response.

[0010] In geriatric and degenerative diseases, iron levels in the brain are inevitably elevated. The oxidative stress of excess iron is related to carcinogenesis. Acute kidney injury (AKI), formerly known as acute renal failure (ARF), is a common and critical illness caused by multiple causes, including ischemia, nephrotoxic drugs, and urinary tract obstruction. A variety of molecular mechanisms have been proposed to induce or aggravate AKI, but ROS- induced renal damage is considered to be one of the key mediators. AKI occurs in approximately 5% of hospitalized patients and 30% of critically ill patients and has high morbidity and mortality. Moreover, 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, few therapeutics have made significant progress in the prevention of AKI. Thus, new targets or better regimens are still urgently needed to prevent AKI as well as to facilitate adaptive repair after the occurrence of AKI. Multiple studies have suggested that ferroptosis is a promising therapeutic target, especially in diseases dominated by kidney tubular necrosis. (Linkermann A. et al. Journal of the American Society of Nephrology. 2014;25(12):2689- 2701.) [0011] It has been universally acknowledged that chelating iron ameliorates and/or prevents ferroptosis. Unfortunately, the only suggested chelators in the literature are Deferoxamine (DESFERAL®) which is approved only for intra- venous and sub-cutaneous administration; 2,2'-bipyridyl (2, 2 '-Dipyridine) which is a highly toxic substance, that shows efficacy in cell cultures; and ciclopirox olamine (LOPROX®) which is a chelating fungicide for topical use and is not approved for systemic use). All of these have safety issues and are not suitable for long term usage to prevent ongoing ferroptosis and have never been evaluated at doses that are not toxic for long term use. Ferroptosis is one of the promising therapeutic targets, especially in diseases dominated by kidney tubular necrosis like ischemic, cisplatin nephrotoxic, and rhabdomyolysis-induced AKI.

[0012] In general, lipid peroxidation inhibitors, such as lysyl oxidase inhibitors, Ferrostatin- 1 and Liproxstatin- 1 have been used to inhibit ferroptosis. In some of these models, iron chelators were investigated long before the detection of ferroptosis. However, those compounds, such as deferoxamine, never made it into the clinical routine, despite considerable effects in ex vivo experiments with kidney tubules. Antioxidants and iron chelators (such as vitamin E and deferoxamine) were also observed to inhibit ferroptosis by reducing iron availability but only in in vitro experiments and rodent models. Chronically administered deferoxamine has been reported to have multiple adverse effects, such as acute respiratory distress syndrome, visual defects, and enhancement of Yersinia enterocolitica infection.

[0013] When used in patients without iron overload, deferoxamine can cause iron deficiency and falls in ferritin concentrations. In the setting of a single dose, flushing, erythema, tachycardia, urticaria, and hypotension caused by rapid administration of deferoxamine are the most serious side effects. Deferoxamine is approved only for intramuscular and intravenous administration in the setting of acute iron intoxication.

[0014] There is a need for systemic administrating iron chelators to a subject, human or animal, in order to alleviate and treat diseases, conditions (including aging) and viral infections that can potentially cause ferroptosis. There are other iron chelators that are commonly used in foods, such as EDTA disodium, EDTA calcium disodium, metaphosphates and others. While these could potentially be used for oral administration, they are generally not absorbed into the body through oral delivery. [0015] Therefore, there is a need for novel formulations comprising a systemically administrable iron chelator and one or more permeating agents that allow the chelator to be absorbed into the body. Especially, during a pandemic caused by a rapidly mutating RNA virus a desirable formulation for inhibiting viral replication in a form that can be readily used to reduce iron levels in a subject and thereby inhibit viral replication and viral load.

SUMMARY OF THE INVENTION

[0016] The instant invention provides a formulation that can be used to control iron levels in the body by regularly replenishing iron chelator in the subject. Diseases and conditions associated with high iron levels such as viral infections and ferroptosis are thereby modulated. In one embodiment, the formulation is a potable liquid such as a ready-to-drink beverage. The novel formulations disclosed herein are useful in controlling viral load in an infected subject and also useful on ferroptosis mediated diseases and conditions such as cancer, and aging.

[0017] Formulations comprising a transport enhancer (such as MSM) and a chelating agent (such as EDTA), for topical applications to ocular, dental and dermal surfaces have been previously disclosed and patented by the present inventors. (WO 2013/166459 by Bhushan et al.; WO 2014/100775 by Bhushan et al.; US Patent No. 9,616,008 by Bhushan et al.).

[0018] The invention provides a formulation comprising: a chelating agent or salts thereof, wherein the chelating agent is suitable for long-term consumption; a permeation enhancer that is methylsulfonylmethane (MSM); one or more inert excipients; and a liquid vehicle or carrier; wherein the chelating agent and the permeation enhancer are present in proportions effective to maintain homeostasis of iron levels in the body when consumed in regular doses, and wherein the percentage of chelator is about 0.0001% to 15% and the percentage of permeation enhancer in the composition is about 0.0001% - 30% by weight, respectively.

[0019] In one aspect of the invention, the formulation is in the form of a concentrate the form of a liquid, a dissolvable solid, an effervescent tablet, a pill or a form that can be readily reconstituted.

[0020] In one aspect of the invention, the formulation is in a liquid form capable of being administered by a route selected from the group consisting of oral, intranasal, inhalation, intravenous, intra-muscular, transdermal, topical, rectal, vaginal, buccal, injection, sublingual, or combination thereof. In a particular aspect the formulation comprises a hydration drip further comprising electrolytes, vitamins, or other nutrients. [0021] In one aspect of the invention, homeostasis of iron levels is sufficient to inhibit replication of an RNA virus such as a coronavirus, a retrovirus, HIV-1, SARS-CoV-2, MERS, SARS, influenza, HTLV-I and HTLV-II in a human or animal subject.

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

[0023] In particular aspects of the invention, the inhibited ferroptosis is associated with a pathologic disease or condition in a subject, including disease or condition related to an organ selected from: heart, central nervous system, liver, gastrointestinal organs, lung, kidney and pancreas.

[0024] In particular aspects of the invention, the inhibited ferroptosis is associated with cancer, aging, inflammation, hearing loss, neurodegenerative diseases, and I/R (Ischemia Reperfusion) injury-related diseases.

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

[0026] In one aspect of the invention, the homeostasis of iron levels is maintained by chelating iron associated with heme-containing proteins selected from hemoglobin, myoglobin and neuroglobin.

[0027] The method involves administering to the subject an effective amount of a formulation composed of a therapeutically effective amount of a chelating agent and an effective transport-enhancing amount of a transport enhancer having the formula (I)

wherein R¹ and R² are independently selected from C2-C6 alkyl, Ci-Ce heteroalkyl, C6-C14 aralkyl, and C2-C12 heteroaralkyl, any of which may be substituted, and Q is S or P.

[0028] The transport enhancing agent can be, for example, methylsulfonylmethane (MSM; also referred to as methylsulfone, dimethylsulfone, and DMSO2).

[0029] The chelating agent can be selected from ethylenediamine tetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), cyclohexanediamine tetraacetic acid (CDTA), hydroxy ethylethylenediamine triacetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTP A), dimercaptopropane sulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylene phosphonic acid (ArPA), citric acid, acetic acid, phosporic acid, pyro phosphates, meta phosphates, malic acid polymers, etc., and acceptable salts thereof, and any combinations thereof.

[0030] Methods for treating or alleviating Covid- 19 by inhibiting SAR-CoV2 replication are disclosed herein.

[0031] Methods for preventing, treating or alleviating diseases or conditions associated with high iron levels in the cell, hyper-ferritinemia and ferroptosis are disclosed herein.

[0032] These and other aspects will become apparent from the following description of the preferred embodiment taken in conjunction with the following drawings, although variations and modifications therein may be affected without departing from the spirit and scope of the novel concepts of the disclosure.

BRIEF DESCRIPTION OF THE DRAWINGS

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

[0034] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0035] Figure 1 illustrates the role ferroptosis has played important roles in multiple system diseases, such as nervous system diseases, heart diseases, liver diseases, gastrointestinal diseases, lung diseases, kidney diseases, pancreatic diseases, and so on. Figure 1 is adopted from Li J. et al. Cell Death and Disease (2020) 11:88.

[0036] Figure 2 illustrates the vicious cycle of aging mechanism, promoted by underlying chronic diseases and ferroptosis. Illustration showing ferroptosis as a catalytic factor for induction of a vicious cycle of accelerated aging and chronic disease progression with underlying imbalance of oxidant and antioxidant defense mechanisms. Figure 1 is adopted from Mazhar M, et al. Cell Death Discovery (2021) 7:149; accessed January 28, 2022 at https://doi.org/10.1038/s41420-021-00553-6. [0037] Figure 3 shows H&E-stained intestinal mucosa 3 days after induction of colitis with TNBS. CONTROL: mucosal architecture is completed, submucosa no cell infiltration. TNBS: colon mucosal structured disorder, submucosa cell infiltration.

TNBS+ME(MSM+EDTA): colon structure a little bit changed; submucosa a few cells infiltration.

[0038] Figure 4 shows H&E-stained intestinal mucosa 3 days after induction of colitis with DSS. CONTROL: mucosal architecture is completed, submucosa no cell infiltration. DSS: colon mucosal ulcer (Arrow), goblet cell depletion and structured disorder; submucosal edema, cell infiltration. DSS+ME(MSM+EDTA): colon structure a little bit changed; submucosa a few cells infiltrated.

[0039] Figure 5 illustrates IHC Stained with ALDH1, Protein-HNE, Protein- Acrolein, Protein-MDA in s DSS model of colitis in rats, showing alleviation of microscopic colon damage with MSM+EDTA (ME) treatment. Upper row is Anti- Protein-HNE staining, Lower row is anti-protein-HNE with DAPI. A Normal control. B: DSS induced colitis. C: DSS induced colitis treated with ME.

[0040] Figure 6 illustrates chronic inflammation with IL-6 as a marker in rats that were dosed ad libitum with normal water or water enriched with MSM and chelator. The MSM/EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal Rats dosed with normal water, NR+ME: Normal Rats + MSM/EDTA drinking water. DR: Diabetic Rats dosed with normal water, DR+ME: Diabetic Rats + MSM/EDTA drinking water.

[0041] Figure 7A shows a low magnification (lOOx) photomicrograph of a pancreatic lobule, 4pm section of formalin-fixed, paraffin-embedded pancreas, H&E stained. A section of the pancreas (A) from normal rat, shows normal endocrine islets of Langerhans in number and size and normal endocrine acinar tissue. (B) from normal rat dosed orally with M+E (MSM + EDTA) also shows normal pancreas islets and acinar tissue. (C) from diabetic rat indicated that pancreatic endocrine islets of Langerhans obviously reduced in number and size. Most of the islets were small, shrunk and inconspicuous. (D) from diabetic rat, dosed orally with M+E had distinctly improved endocrine islets of Langerhans in number and size and there was no shrinkage of acinar tissue.

[0042] Figure 7B shows a higher magnification photomicrograph (400x) of pancreatic endocrine islets, 4pm sections of formalin-fixed, paraffin-embedded pancreas, H&E stained, (A) An endocrine islet in the pancreas of a normal rat shows interspersed cells in lightly stained exocrine acinar glands, spherical clusters of cells without ducts, and acini. (B). An endocrine islet in pancreas section from normal rat, dosed orally with ME, shows that the histology and morphology were not significantly changed, (C) An endocrine islet in the pancreas of a diabetic rat shows that the islet of Langerhans shrink and become small and inconspicuous (sclerosis of islet, reduction of the cell’s cytoplasm) and show the presence of inter-acinar pancreatitis as evident from leukocytic infiltration in the islets. (D). An endocrine islet of diabetic rat, dosed orally with ME shows that the islet of Langerhans had mild shrinkage and negligible leukocytic infiltration.

DETAILED DESCRIPTION OF THE INVENTION

[0043] The terms used in this specification generally have their ordinary meanings in the art, within the context of the invention, and in the specific context where each term is used. Certain terms that are used to describe the invention are discussed below, or elsewhere in the specification, to provide additional guidance to the practitioner regarding the description of the invention. For convenience, certain terms may be highlighted, for example using italics and/or quotation marks. The use of highlighting has no influence on the scope and meaning of a term; the scope and meaning of a term is the same, in the same context, whether or not it is highlighted. It will be appreciated that same thing can be said in more than one way. Consequently, alternative language and synonyms may be used for any one or more of the terms discussed herein, nor is any special significance to be placed upon whether or not a term is elaborated or discussed herein. Synonyms for certain terms are provided. A recital of one or more synonyms does not exclude the use of other synonyms. The use of examples anywhere in this specification including examples of any terms discussed herein is illustrative only, and in no way limits the scope and meaning of the invention or of any exemplified term. Likewise, the invention is not limited to various embodiments given in this specification.

[0044] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the invention, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the invention.

[0045] When referring to a formulation component, it is intended that the term used, e.g., "agent," encompass not only the specified molecular entity but also its pharmaceutically acceptable analogs, including, but not limited to, salts, esters, amides, prodrugs, conjugates, active metabolites, and other such derivatives, analogs, and related compounds.

[0046] The terms "treating" and "treatment" as used herein refer to the administration of an agent or formulation to a clinically symptomatic individual human or animal afflicted with an adverse condition, disorder, or disease, so as to affect a reduction in severity and/or frequency of symptoms, eliminate the symptoms and/or their underlying cause, and/or facilitate improvement or remediation of damage. The terms "preventing" and "prevention" refer to the administration of an agent or composition to a clinically asymptomatic individual who is susceptible to a particular adverse condition, disorder, or disease, and thus relates to the prevention of the occurrence of symptoms and/or their underlying cause. Unless otherwise indicated herein, either explicitly or by implication, if the term "treatment" (or "treating") is used without reference to possible prevention, it is intended that prevention be encompassed as well, such that "a method for the treatment of gingivitis" would be interpreted as encompassing "a method for the prevention of gingivitis."

[0047] "Optional" or "optionally present" - as in an "optional substituent" or an "optionally present additive" means that the subsequently described component (e.g., substituent or additive) may or may not be present, so that the description includes instances where the component is present and instances where it is not.

[0048] By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, e.g., the material may be incorporated into a formulation of the invention without causing any 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 excipient, it is implied that the excipient has met the required standards of toxicological and manufacturing testing and/or that it is included on the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration. As explained in further detail infra, "pharmacologically active" (or simply "active") as in a "pharmacologically active" derivative or analog refers to derivative or analog having the same type of pharmacological activity as the parent agent. The terms "treating" and "treatment" as used herein refer to reduction in severity and/or frequency of symptoms, elimination of symptoms and/or underlying cause, prevention of the occurrence of symptoms and/or their underlying cause, and improvement or remediation of an undesirable condition or damage. Thus, for example, "treating" a subject involves prevention of an adverse condition in a susceptible individual as well as treatment of a clinically symptomatic individual by inhibiting or causing regression of the condition. The term "chelating agent" (or "active agent") refers to any chemical compound, complex or composition that exhibits a desirable effect in the biological context, i.e., when administered to a subject or introduced into cells or tissues in vitro. The term includes pharmaceutically acceptable derivatives of those active agents specifically mentioned herein, including, but not limited to, salts, esters, amides, prodrugs, active metabolites, isomers, analogs, crystalline forms, hydrates, and the like. When the term "chelating agent" is used, or when a particular chelating agent is specifically identified, it is to be understood that pharmaceutically acceptable salts, esters, amides, prodrugs, active metabolites, isomers, analogs, etc. of the agent are intended as well as the agent per se.

[0049] By an "effective" amount or a "therapeutically effective" amount of an active agent is meant a nontoxic but sufficient amount of the agent to provide a beneficial effect. The amount of active agent that is "effective" will vary from subject to subject, depending on the age and general condition of the individual, the particular active agent or agents, and the like. Unless otherwise indicated, the term "therapeutically effective" amount as used herein is intended to encompass an amount effective for the prevention of an adverse condition and/or the amelioration of an adverse condition, i.e., in addition to an amount effective for the treatment of an adverse condition.

[0050] As will be apparent to those of skill in the art upon reading this invention, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present invention. Any recited method can be carried out in the order of events recited or in any other order that is logically possible.

[0051] Unless otherwise indicated, the invention is not limited to specific formulation components, modes of administration, chelating agents, manufacturing processes, or the like, as such may vary. [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 the case of conflict, the present document, including definitions will control.

Role of Iron in Covid- 19

[0053] Iron overload is increasingly implicated as a contributor to the pathogenesis of CO VID- 19. Indeed, several of the manifestations of CO VID- 19, such as inflammation, hypercoagulation, hyperferritinemia, and immune dysfunction are also reminiscent of iron overload. Although iron is essential for all living cells, free unbound iron, resulting from iron dysregulation and overload, is very reactive and potentially toxic due to its role in the generation of reactive oxygen species (ROS). ROS react with and damage cellular lipids, nucleic acids, and proteins, with consequent activation of either acute or chronic inflammatory processes implicated in multiple clinical conditions.

[0054] CO VID- 19 manifests itself in many complications as well as physiological and biochemical alterations. These include, but are not limited to, Acute Respiratory Distress Syndrome (ARDS), high concentrations of proinflammatory CD4 T cells and cytotoxic granules CD8 T, massive release of cytokines (cytokine storm), increased coagulation state, hemoglobin damage and dysregulation of iron homeostasis including iron overload. (Habib, 2021).

[0055] After 20 months of global circulation, basal lineages of SARS-CoV-2 have been almost completely replaced by derived, variant lineages. These lineages are classified by the WHO as variants of concern (VOCs) or variants of interest (VOIs) based on genetic, phenotypic and epidemiological differences. Despite possessing some proof-reading capacity (a relatively rare function for an RNA virus), SARS-CoV-2 has been accumulating roughly 24-25 substitutions per year. Rigorous quantification of the evolutionary process shows that the observed success of variant viruses is a result of adaptive, not neutral, evolution. (Kistler KE, et al. preprint accessed January 28, 2022 at: https://doi.org/10.1101/2021.09.ll.459844).

[0056] Iron plays two roles in Covid-19 disease and the severity of its symptoms, (a) All coronaviruses (including SARS, MERS, SARS-Cov2) depend upon iron for their replication (Shi 2005, Liu 2020, Habib 2021), and under low iron conditions, the replication slows down (Menshawey 2020). (b) Co vid patients often suffer from iron overload, resulting in Ferroptosis, leading to damage to multiple organs. (Chen 2021, Yang 2020, Edeas 2020). [0057] The main stores of iron in the body are cells containing hemoglobin, myoglobin, and neuroglobin. Iron is an essential element for blood production. Over 70 percent of the body's iron is found in the red blood cells in a heme-containing protein called hemoglobin, in muscle cells as a heme-containing protein called myoglobin and in nerve cells as a heme-containing protein called neuroglobin. About 6 percent of body iron is a component of certain proteins, essential for respiration and energy metabolism, and as a component of enzymes involved in the synthesis of collagen and some neurotransmitters. Iron also is needed for proper immune function. About 20-25 percent of the iron in the body is stored as ferritin, which is found in cells and circulating blood.

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

[0059] The main entry point of SARS-CoV-2 into cells by attachment to the angiotensinconverting enzyme 2 (ACE2), which is attached to the outer surface of the cell membranes of cells in the lungs, arteries, heart, kidney and intestines. The binding of the spike SI protein of SARS-CoV and SARS-CoV-2 to the enzymatic domain of ACE2 on the surface of cells results in endocytosis and the translocation of both the virus and the enzyme into endosomes located within cells. Liu, et al. suggest that the strong affinity of SARS-CoV-2 with human ACE2 molecules indicates that the key pathogenic molecular step of COVID-19 is to attack ACE2 positive cells. More than 80% of ACE2 receptors are expressed in a small population of type II alveolar cells (AT2).

[0060] Iron is very tightly regulated in the body. Sufficient intracellular iron levels support coronavirus replication, whereas intracellular iron deficiency undermines its replication process by interfering with viral transcription, translation, assembly, and exocytosis. The main stores of iron in the body are cells with heme-containing proteins such as hemoglobin, myoglobin, and neuroglobin. Iron is an essential element for blood production.

[0061] In order to replicate, a coronavirus needs iron and therefore infecting cells containing iron is productive for its replication. However, in order for Covid virus to enter a cell the cell has to have ACE2 receptors. Red blood cells which have hemoglobin do not have ACE2 receptors on their surface. There might be another pathway on the surface of red blood cells that can interact with SI spike proteins of SARS-CoV2. One such possible pathway is through the interaction with the red blood cell (RBC) Band3 surface protein. (Cosic I, et al., Appl. Sci. 2020, 10, 4053; doi:10.3390/appl0114053). However, the virus does infect other iron-rich cells containing ACE2 surface receptors, including muscle cells (myoglobin) and nerve cells (neuroglobin).

[0062] Hemoglobin, myoglobin and neuroglobin lose their capacity to bind oxygen and hinders its delivery to major organs, which results in rapid multi-organ failures. Moreover, the free iron released into the circulation may result in iron overload causing oxidative damage to the lungs and other organs. Iron overload may also result in inflammation and immune dysfunction. These dictate increased uptake and storage of iron in iron-binding proteins. Indeed, this notion is supported by the increased ferritin (the iron storage molecule in the body) concentrations in the circulation of COVID-19 patients. Amplified iron load leads to increased blood viscosity with recurrent and diffuse macro and micro circulatory thrombosis; this may explain the cause of unexpected deterioration and death in some cases.

[0063] Iron dependence of viral replication and modulation of host iron metabolism by RNA viruses, signifies the importance of cellular iron homeostasis in viral life cycle and suggests the utility of iron chelation strategy in treating viral infections. One strategy is to deplete iron directly by chelators which have strong and selective affinity with iron ions. Some iron chelators have been approved by U.S. Food and Drug Administration for clinical use, such as deferoxamine (DFO, DESFERAU®), deferiprone (DFP, FERRIPROX®), and deferasirox (ICE670, EXJADE®). Iron chelators can bind free iron or remove iron from iron-containing proteins. Deferasirox is a membrane-permeable iron chelator and the first oral medication approved by the Food and Drug Administration (FDA) for chronic iron overload in the body caused by multiple blood transfusions. Treatment with higher doses of DFP has been shown to prolong the survival of AIDS patients after HIV-1 infection. Ciclopirox is a synthetic broad- spectrum antifungal drug that binds divalent cations like Fe³⁺. 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 stoichiometry of 1 : 1.

[0064] Iron chelation drugs are able to bind free iron but they can also remove iron from iron-containing proteins, which means that iron chelation can have an anti-ferritin effect. In fact, deferoxamine increases degradation of ferritin by lysosomes. While iron chelation can play a critical role in the control and treatment of CO VID- 19, the approved iron chelators are not suitable for long term use. The most used approved iron chelator (Deferoxamine, DFO) has a downside of degrading ferritins (De Domenico I, et al., BLOOD, 12 November 2009, Volume 114, Number 20 2009; Abobaker A, European Journal of Clinical Pharmacology (2021) 77:267-268), and potentially worsening the symptoms. However, food grade chelators, and in particular EDTA do not have that effect on ferritin.

Ferroptosis

[0065] The upstream inducers of ferroptosis can be divided into two categories (biological versus chemical) and activate two major pathways (the extrinsic/transporter versus the intrinsic/enzymatic pathways). Ferroptosis does not have the morphological characteristics of typical necrosis, such as swelling of the cytoplasm and organelles and rupture of the cell membrane, nor does it have the characteristics of traditional cell apoptosis, such as cell shrinkage, chromatin condensation, formation of apoptotic bodies and disintegration of the cytoskeleton. In contrast to autophagy, ferroptosis does not have the formation of classical closed bilayer membrane structures (autophagic vacuoles). Morphologically, ferroptosis mainly manifests as obvious shrinkage of mitochondria with increased membrane density and reduction in or vanishing of mitochondrial cristae, which is a different process from other modes of cell death.

[0066] Current knowledge indicates that ferroptosis occurs during various pathophysiological processes of the body, including degenerative diseases of the central nervous system, the antiviral immune response, arteriosclerosis, acute kidney injury, diabetes, and ischemiareperfusion injury. (Oxid Med Cell Longevity. 2019; 2019:8010614; published online 2019 Oct 31). Ferroptosis is characterized by the accumulation of membrane lipid peroxidation products and the consumption of plasma membrane polyunsaturated fatty acids. This kind of cell death can be induced by specific small molecules such as erastin and RAS-selective lethal 3 (RSL3). Ferroptosis has been reported to participate in various pathological processes of the brain, kidney, liver, and heart diseases. (Sheng X., et al. Physical Chemistry Chemical Phy sics . 2017 ; 19(20) : 13153-13159) .

[0067] Glutathione (L-glutamyl-L-cysteinylglycine [GSH]) depletion can lead to irondependent accumulation of reactive oxygen species (ROS), especially lipid ROS, which are themselves sufficient to kill cells. (Dixon, 2012). Iron metabolism and lipid peroxidation signaling are thought to be central mediators of ferroptosis. Circulating iron exists in the form of ferric iron (Fe3+) bound to transferrin, thereby reducing free iron levels. Excessive iron can lead to the production of ROS that mediate ferroptosis. The biological properties of ferroptosis are characterized by iron and ROS aggregation. When ferroptosis occurs, the cell membrane ruptures and get blistered, the cell nucleus become lacking of chromatin condensation, the mitochondria decrease, mitochondria size decreases, the density of the bilayer membrane increases, the mitochondrial cristae decrease or disappear, and the mitochondrial outer membrane ruptures as observed under electron microscopy.

[0068] Ferroptosis is distinct from apoptosis and necroptosis as shown by its independent mediation in the absence of key effector of apoptosis and necroptosis i.e., BAX, BAK, caspase; mixed lineage kinase domain like protein (MLKL), and receptor interacting serine/threonine kinases (RIPK1 and RIPK3). Although, ferroptosis has a cancer protective effect on uncontrolled tumor cells, however, it is a possible culprit underlying pathogenesis of various diseases and can be a hidden cause of many unidentified disease mechanisms. Despite rigorous research in the field of ferroptosis, still a lot remain to be explored to fully understand its mechanism and its role in the physiological and pathological conditions. So far, the mechanism of ferroptosis as described by various researchers can be simply comprised of four steps i.e., (i) inactivation of cysteine/glutathione antiporter system, (ii) depletion of glutathione and GPx4, (iii) excessive production of lipid ROS and (iv) excessive cellular iron accumulation.

[0069] Ferroptosis spreads in a paracrine manner via signals not clearly determined yet, but may include the toxic end products of lipid peroxidation, 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA) which are stable and react with biological macromolecules, and can affect distant sites from its origin. Electron lucent nucleus is found to be a unique feature and hallmark of ferroptosis through transmission electron microscopy. The morphological characteristics induced in a cell upon ferroptosis are distinct from apoptosis and mainly influence mitochondria. Dixon et al. observed mitochondrial shrinkage and dysfunction that indicate ferroptosis is directly linked with cellular energetics and mitochondrial dysfunction leading to alterations and lack of cellular energy and eventually cell death. The classic profile of ferroptosis is regulated cell death due to iron dependent lipid peroxidation, which can be ameliorated by iron chelators and lipid antioxidants, and the culprit of lipid peroxidation is generally believed to be ROS that react with polyunsaturated fatty acids (PUFAs) of the membrane and induce lipid peroxidation. Several ROS producing routes have been proposed, however, the detailed mechanism of iron induced ROS remains unclear.

[0070] Ferroptosis has played important roles in various diseases of different organs as shown in Figure 1. (the original Fig. 1 in Li J. et al. Cell Death and Disease (2020) 11:88; accessed January 28, 2022 at https://doi.org/10.1038/s41419-020-2298-2). Ferroptosis also is involved in the vicious cycle of the aging process as shown in Fig 2 (from the original Fig. 1 found in Mazhar M, et al. Cell Death Discovery (2021) 7:149; accessed January 28, 2022 at https://doi.org/10.1038/s41420-021-00553-6).

[0071] It has been universally acknowledged that chelating iron ameliorates/prevents ferroptosis. However, the only suggested chelators in the literature are Deferoxamine (DESFERAL®) which is approved only for intra- venous and sub-cutaneous administration; 2,2'-bipyridyl (2, 2 '-Dipyridine) which is a highly toxic substance, that shows efficacy in cell cultures; and ciclopirox olamine (LOPROX®) which is a chelating fungicide for topical use, and is not approved for systemic use). All of these have safety issues and are not suitable for long term usage to prevent ongoing ferroptosis and have never been evaluated at doses that are not toxic for long term use.

[0072] Some iron chelators are commonly used in foods, such as EDTA disodium, EDTA calcium disodium, metaphosphates and others. While they are generally not absorbed into the body through oral delivery, these compounds could potentially be used for oral administration in order to chelate iron and prevent or ameliorate ferroptosis.

Chelating agent:

[0073] Chelation is a chemical combination with a metal in complexes in which the metal is part of a ring. An organic ligand is called a chelator or chelating agent, the chelate is a metal complex. The larger number of ring closures to a metal atom the more stable is the compound. The stability of a chelate is also related to the number of atoms in the chelate ring. Monodentate ligands which have one coordinating atom like H2O or NH3 are easily broken apart by other chemical processes, whereas polydentate chelators, donating multiple binds to metal ion, provide more stable complexes. Chlorophyll, a green plant pigment, is a chelate that consists of a central magnesium atom joined with four complex chelating agents (pyrrole ring). Heme is an iron chelate which contains iron (II) ion in the center of the porphyrin. Chelating agents offers a wide range of sequestrants to control metal ions in aqueous systems. By forming stable water-soluble complexes with multivalent metal ions, chelating agents prevent undesired interaction by blocking normal reactivity of metal ions. EDTA (ethylenediamine tetraacetate) is a good example of common chelating agents which have nitrogen atoms and short chain carboxylic groups. [0074] For the purposes of this invention that is safely ingestible is suitable for formulating a composition that can be consumed by a subject in need of iron chelation. Examples of chelators of iron and calcium include, but are not limited to, Diethylene triamine pentaacetic acid (DTP A), ethylene diamine tetraacetic acid (EDTA), nitrilotriacetic acid (NT A), 1,3- propylene diamine tetraacetic acid (PDTA), Ethylene diamine disuccinic acid (EDDS), and ethylene glycol tetraacetic acid (EGTA). Any suitable chelating agent known in the art, which is biologically safe and able to chelate iron, calcium or other metals, is suitable for the invention.

[0075] The concentration of the chelator must be sufficient to allow the chelating agent to be transferred across the cell membrane in order to access intracellular iron. Thus, the concentration of the chelator will be decided by the thickness of the cell membrane in the animal or human subject who is ingesting the formulation. For example, in exemplary evaluations, rats were dosed with ad libitum drinking water with 26 ppm (0.0026%) EDTA disodium which, adjusted for humans, would be equivalent to 260 ppm (0.026%). In some embodiments, the transport enhancer may be present in a formulation of the invention in an amount that ranges from 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 compounds that coordinate to or form complexes with a divalent or polyvalent metal cation, thus serving as a sequestrant of such cations. Accordingly, the term "chelating agent" herein includes not only divalent and polyvalent ligands (which are typically referred to as "chelators") but also monovalent ligands capable of coordinating to or forming complexes with the metal cation.

[0077] Suitable biocompatible chelating agents useful in conjunction with the present invention include, without limitation, monomeric polyacids such as EDTA, cyclohexanediamine tetraacetic acid (CDTA), hydroxy ethylethylenediamine triacetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTP A), dimercaptopropane sulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylene phosphonic acid (ATP A), citric acid, pharmaceutically acceptable salts thereof, and combinations of any of the foregoing. Other exemplary chelating agents include: phosphates, e.g., pyrophosphates, tripolyphosphates, and hexametaphosphates, malic acid polymers, and others.

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

[0079] EDTA has been widely used as an agent for chelating metals in biological tissue and blood, and has been suggested for inclusion in various formulations. For example, U.S. Pat. No. 6,348,508 to Denick Jr. et al. describes EDTA as a sequestering agent to bind metal ions. In addition to its use as a chelating agent, EDTA has also been widely used as a preservative in place of benzalkonium chloride, as described, for example, in U.S. Pat. No. 6,211,238 to Castillo et al. U.S. Pat. No. 6,265,444 to Bowman et al. discloses use of EDTA as a preservative and stabilizer. However, EDTA has generally not been applied topically in any significant concentration formulations because of its poor penetration across biological membranes.

[0080] Among the chelating/sequestering materials which may be included in the compositions there may be mentioned biocompatible chelating agents include, without limitation, monomeric polyacids such as EDTA, cyclohexanediamine tetraacetic acid (CDTA), hydroxyethylethylenediamine triacetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTP A), dimercaptopropane sulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylene phosphonic acid (ATP A), citric acid, pharmaceutically acceptable salts thereof, and combinations of any of the foregoing.

[0081] Other exemplary chelating agents include: phosphates, e.g., pyrophosphates, tripolyphosphates, and hexametaphosphates. Other exemplary chelating agents include: phosphates, e.g., pyrophosphates, tripolyphosphates, and hexametaphosphates; chelating antibiotics such as chloroquine and tetracycline; nitrogen-containing chelating agents containing two or more chelating nitrogen atoms within an imino group or in an aromatic ring (e.g., diimines, 2,2'-bipyridines, etc.); and polyamines such as cyclam (1,4,7,11- tetraazacyclotetradecane), N-(CI-C3O alkyl)-substituted cyclams (e.g., hexadecyclam, tetramethylhexadecy Icy clam), diethylenetriamine (DETA), spermine, diethylnorspermine (DENSPM), diethylhomo-spermine (DEHOP), deferoxamine (N'-{5- [Acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4- oxobutanoyl } amino)pentyl]-N-hydroxysuccinamide, or N'- [5-(Acetyl-hydroxy- amino)pentyl]-N-[5-[3-(5-aminopentyl-hydroxy-carbamoyl) propanoylamino]pentyl]-N- hydroxy-butane diamide); also known as desferrioxamine B, desferoxamine B, DFO-B, DFOA, DFB or desferal), deferiprone, pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), ethane-l,2-bis(N-l-amino-3-ethylbutyl-3- thiol).

[0082] Additional, suitable biocompatible chelating agents which may be useful for the practice of the current disclosure include EDTA-4-aminoquinoline conjugates such as ([2- (Bis-ethoxycarbonylmethyl-amino)-ethyl]-{[2-(7-chloro-quinolin-4-ylamino)- ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([2-(Bis-ethoxycarbonylmethyl- amino)-propyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([3-(Bis-ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinolin-4- ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([4-(Bis- ethoxycarbonylmethyl-amino)-butyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]- methyl}-amino)-acetic acid ethyl ester, ([2-(Bis-ethoxymethyl-amino)-ethyl]-{[2-(7-chloro- quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([2-(Bis- ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}- amino)-acetic acid ethyl ester, ([3-(Bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinolin- 4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([4-(Bis-ethoxymethyl- amino)-butyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester as described in Solomon et al., Med. Chem. 2: 133-138, 2006.

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

[0084] In some embodiments, the chelating agents are selected from the tetrasodium salt of iminodisuccinic acid (Baypure® CX100; LANXESS GMBH (previously Bayer Chemicals) Leverkusen, DE) or salts of poly-asparatic acid (Baypure® DS 100; LANXESS GMBH, Leverkusen, DE). In some embodiments, the chelating agents are tetra sodium salts of L- glutamic acid N,N-diacetic acid (GLDA - Dissolvine®, AkzoNobel, Netherlands).

[0085] In some embodiments, the chelating agent incorporated in the formulation is a prochelator. A prochelator is any molecule that is converted to a chelator when exposed to the appropriate chemical or physical conditions. For example, BSIH (isonicotinic acid [2- (4,4,5,5-tetramethyl-[l,3,2] dioxaborolan-2-yl)-benzylidene]-hydrazide) prochelators are converted by hydrogen peroxide into SIH (salicylaldehyde isonicotinoyl hydrazone) iron- chelating agents that inhibit iron-catalyzed hydroxyl radical generation. [0086] The inactivated metal ion sequestering agent is sometimes referred to herein as a "prochelator," although sequestration of metal ions can involve sequestration and complexation processes beyond the scope of chelation per se. The term "prochelator" is analogous to the term "prodrug" insofar as a prodrug is a therapeutically inactive agent until activated in vivo, and the prochelator, as well, is incapable of sequestering metal ions until activated in vivo.

Transport Enhancer:

[0087] The transport enhancer is selected to facilitate the transport of a chelating agent through the tissues, extra-cellular matrices, and/or cell membranes of a body. An "effective amount" of the transport enhancer represents an amount and concentration within a formulation of the invention that is sufficient to provide a measurable increase in the penetration of a chelating agent through one or more of the sites of oral cavity or teeth in a subject than would otherwise be the case without the inclusion of the transport enhancer within the formulation.

[0088] The concentration of the transport enhancer must be sufficient to allow the chelating agent to be transferred across the cell membrane. Thus, the concentration or relative amount of the transport enhancer will be decided by the thickness of the cell membrane in the animal or human subject who is ingesting the formulation. In some embodiments, the concentration of MSM in the present formulations is in the range of about 0.0001 wt. % to 30 wt. %, or from about 0.01 wt.% to about 0.10, 1, 5, 10, 20, 30 wt.%, and preferably between about 0.01 wt. % to 1.0 wt. %.

[0089] The transport enhancer is generally of the formula (I)

(I)

(?)

R? — Q — R²

O wherein R¹ and R² are independently selected from C2-C6 alkyl, Ci-Ce heteroalkyl, C6-C14 aralkyl, and C2-C12 heteroaralkyl, any of which may be substituted, and Q is S or P. Compounds wherein Q is S and R¹ and R² are C1-C3 alkyl are preferred, with methylsulfonylmethane (MSM) being the optimal transport enhancer. [0090] The phrase "having the formula" or "having the structure" is not intended to be limiting and is used in the same way that the term "comprising" is commonly used. With respect to the above structure, the term "alkyl" refers to a linear, 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 and the like. If not otherwise indicated, the term "alkyl" includes unsubstituted and substituted alkyl, wherein the substituents may be, for example, halo, hydroxyl, sulfhydryl, alkoxy, acyl, etc. The term "alkoxy" intends an alkyl group bound through a single, terminal ether linkage; that is, an "alkoxy" group may be represented as -O-alkyl where alkyl is as defined above. The term "aryl" refers to an aromatic substituent containing a single aromatic ring or multiple aromatic rings that are fused together, directly linked, or indirectly linked (such that the different aromatic rings are bound to a common group such as a methylene or ethylene moiety). Preferred aryl groups contain 5 to 14 carbon atoms. Exemplary aryl groups contain one aromatic ring or two fused or linked aromatic rings, e.g., phenyl, naphthyl, biphenyl, diphenylether, diphenylamine, benzophenone, and the like. "Aryl" includes unsubstituted and substituted aryl, wherein the substituents may be as set forth above with respect to optionally substituted "alkyl" groups. The term "aralkyl" refers to an alkyl group with an aryl substituent, wherein "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, without limitation, benzyl, 2-phenyl-ethyl, 3 -phenylpropyl, 4-phenyl-butyl, 5 -phenyl -pentyl, 4-phenylcyclohexyl, 4-benzylcyclohexyl, 4- phenylcyclohexylmethyl, 4-benzylcyclohexylmethyl, and the like. The term "acyl" refers to substituents having the formula -(CO)-alkyl, -(CO)-aryl, or -(CO)-aralkyl, wherein "alkyl," "aryl, and "aralkyl" are as defined above. The terms "heteroalkyl" and "heteroaralkyl" are used to refer to heteroatom-containing alkyl and aralkyl groups, respectively, i.e., alkyl and aralkyl groups in which one or more carbon atoms is replaced with an atom other than carbon, e.g., nitrogen, oxygen, sulfur, phosphorus or silicon, typically nitrogen, oxygen or sulfur.

[0091] The formulation also includes an effective amount of a transport enhancer that facilitates penetration of the formulation components through cell membranes, tissues, and extracellular matrices. The "effective amount" of the transport enhancer represents a concentration that is sufficient to provide a measurable increase in penetration of one or more of the formulation components through membranes, tissues, and extracellular matrices as just described. Suitable transport enhancers include, by way of example, methylsulfonylmethane (MSM; also referred to as methyl sulfone), combinations of MSM with dimethylsulfoxide (DMSO), or a combination of MSM and, in a less preferred embodiment, DMSO, with MSM particularly preferred. DMSO, a transport enhancer but essentially a solvent, is not particularly suitable for formulations according to this invention. DMSO works as a highly potent solvent and therefore a carrier of its solutes. In contrast, MSM works in a totally different manner by forming hydrogen bonds with select molecules and changing their charge characteristics of the target molecule allowing the target molecule to get through charged barriers like biologic membranes.

[0092] There are differences in chemical structures between MSM and DMSO. Methylsulfonylmethane (MSM) is an organosulfur compound with the formula (CH3)2SO2. It is also known by several other names including DMSO2, methyl sulfone, and dimethyl sulfone. This colorless solid feature a sulfonyl functional group and is considered relatively inert chemically. MSM has the structure:

Dimethyl sulfoxide (DMSO) on the other hand is an organosulfur compound with the formula (CH hSO. This colorless liquid is a widely-used polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible in a wide range of organic solvents as well as water. DMSO has the structure:

[0093] MSM is an odorless, highly water-soluble (34% w/v @ 79° F.) white crystalline compound with a melting point of 108-110° C. and a molecular weight of 94.1 g/mol. MSM serves as a multifunctional agent herein, insofar as the agent not only increases cell membrane permeability, but also acts as a "transport facilitating agent" (TFA) that aids in the transport of one or more formulation components to oral tissues. Furthermore, MSM per se provides medicating effects, and can serve as an anti-inflammatory agent as well as an analgesic. MSM also acts to improve oxidative metabolism in biological tissues, and is a source of organic sulfur, which assists in the reduction of scarring. MSM additionally possesses unique and beneficial solubilizing properties, in that it is soluble in water, as noted above, but exhibits both hydrophilic and hydrophobic properties because of the presence of polar S=O groups and nonpolar methyl groups. The molecular structure of MSM also allows for hydrogen bonding with other molecules, i.e., between the oxygen atom of each S=O group and hydrogen atoms of other molecules, and for formation of van der Waal associations, i.e., between the methyl groups and nonpolar (e.g., hydrocarbon) segments of other molecules.

Formulations

[0094] A variety of means can be used to formulate the compositions of the invention. Techniques for formulation and administration may be found in "Remington: The Science and Practice of Pharmacy," Twenty Third Edition, Adeboye Adejare, editor-in-chief; Academic Press; 23rd edition (November 13, 2020). For human or animal administration, preparations should meet sterility, pyrogenicity, general safety and purity standards comparable to those required by the FDA. Administration of the pharmaceutical formulation can be performed in a variety of ways, some of which are as described herein.

[0095] Other possible additives for incorporation into the formulations that are at least partially aqueous include, without limitation, thickeners, isotonic agents, buffering agents, and preservatives, providing that any such excipients do not interact in an adverse manner with any of the formulation's other components. It should also be noted that preservatives are not generally necessarily in light of the fact that the selected chelating agent itself serves as a preservative.

[0096] The chelator and the permeation agent are dissolved in a solvent selected from a nonlimiting list of solvents that may be employed: 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, wherein the emulsifier is selected from the group consisting of gum arabic, modified starch, pectin, xanthan gum, gum ghatti, gum tragacanth, fenugreek gum, mesquite gum, mono-glycerides and di-glycerides of long chain fatty acids, sucrose monoesters, sorbitan esters, polyethoxylated glycerols, stearic acid, palmitic acid, mono-glycerides, di-glycerides, propylene glycol esters, lecithin, lactylated mono- and di-glycerides, propylene glycol monoesters, polyglycerol esters, diacetylated tartaric acid esters of mono- and di-glycerides, citric acid esters of monoglycerides, stearoyl-2-lactylates, polysorbates, 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, furcellaran, Gellan gum, psyllium, curdlan, konjac mannan, agar, and cellulose derivatives, or combinations thereof.

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

[0099] In some embodiments, 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, dexibup6fen, dexketoprofen, diclofenac, diflunisal, droxicam, etodolac, etoricoxib, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen, indomethacin, isoxicam, ketoprofen, ketorolac, licofelone, lomoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimesulide, oxaprozin, parecoxib, phenylbutazone, piroxicam, rofecoxib, salsalate, sulindac, tenoxicam, tolfenamic acid, tolmetin, or valdecoxib.

[00100] In some embodiments, the ready-to-drink beverage comprises infusions of tea leaves, coffee beans, or cocoa powder,

[0100] Any suitable isotonic agents and buffering agents commonly used in oral formulations may be used, providing 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 formulation is a ready-to-drink beverage selected from the group consisting of noncarbonated beverage, a carbonated beverage, a cola, a root beer, a fruit-flavored beverage, a citrus -flavored beverage, a fruit juice, a fruit-containing beverage, a vegetable juice, a vegetable containing beverage, a tea, a coffee, a dairy beverage, a protein containing beverage, a shake, a sports drink, an energy drink, and a flavored water.

[0102] In preferred embodiments, the pharmaceutical formulation is administered in the form of an orally consumable liquid. However, the formulation may be available as a liquid concentrate, a dissolvable solid, an effervescent tablet or any form that can be readily reconstituted into a drinkable liquid. Concentrates can be in unit dosage form suitable for single administration of a precise dosage. Suitable pharmaceutical formulations and dosage forms may be prepared using conventional methods known to those in the field of pharmaceutical formulation and described in the pertinent texts and literature, e.g., in Remington: The Science and Practice of Pharmacy, cited previously herein.

[0103] The chelating agent may be administered, if desired, in the form of a salt, ester, crystalline form, hydrate, or the like, provided it is pharmaceutically acceptable. Salts, esters, etc. may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry and described, for example, by March's Advanced Organic Chemistry: Reactions, Mechanisms and Structure, 6th Ed. Smith MB and March J, eds. (Wiley- Interscience, 2007).

[0104] The amount of chelating agent administered will depend on a number of factors and will vary from subject to subject and depend on the particular chelating agent, the particular disorder or condition being treated, the severity of the symptoms, the subject's age, weight and general condition, and the judgment of the prescribing physician. The term "dosage form" denotes any form of a pharmaceutical composition that contains an amount of chelating agent and transport enhancer sufficient to achieve a therapeutic effect with a single administration or multiple administrations. The frequency of administration that will provide the most effective results in an efficient manner without overdosing will vary with the characteristics of the particular active agent, including both its pharmacological characteristics and its physical characteristics, such as hydrophilicity.

[0105] The formulations may also include conventional additives such as solvents, flavoring agents, antioxidants, fragrance, colorant, stabilizers, surfactants, and the like. Other agents may also be added, such as antimicrobial agents, to prevent spoilage upon storage, i.e., to inhibit growth of microbes such as yeasts and molds. Suitable antimicrobial agents are typically selected from the methyl and propyl esters of p-hydroxybenzoic acid (i.e., methyl and propyl paraben), sodium benzoate, sorbic acid, imidurea, and combinations thereof.

[0106] The pharmaceutical dosage forms suitable for consumption include aqueous solutions comprising the active ingredients. In all cases, the ultimate dosage form should be a fluid stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like) and suitable mixtures thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. [0107] The treatment regimen will depend on a number of factors that may readily be determined, such as severity of the condition and responsiveness of the condition to be treated, but will normally be one or more treatments per day. Treatment generally comprises consumption of one or more drinking liquids comprising an iron chelator and a permeation enhancer. The course of treatment lasts from a day or several days to several months, or until a consistent desirable level of iron ions in the body or a significant diminution of a ferroptotic condition is achieved.

[0108] The compositions of the invention may further include additional drugs or excipients as appropriate for the indication. In one aspect of the embodiment, the pharmaceutical composition further comprises a therapeutically effective amount of at least one antimicrobial or antifungal agent. In a more specific aspect, the antimicrobial agent is an antibiotic.

EXAMPLES

[0109] The following examples are put forth so as to provide those skilled in the art with a complete invention and description of how to make and use embodiments in accordance with the invention, and are not intended to limit the scope of what the inventors regard as their discovery. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.

[0110] In order to assess the ability of formulations according to the instant invention, rats were dosed with drinking water ad libitum with 26 ppm (0.0026%) EDTA disodium.

Adjusted for humans, this would be equivalent to 260 ppm (0.026%) ad libitum. Since ad libitum consumption is not possible for humans, upward adjustment to 500 to 1000 ppm chelator up to 0.5% to obtain equivalent results. For animals where drinking ad libitum is achievable 10 ppm or higher would obtain equivalent results depending on the animal's weight and size.

[0111] Experiments were performed on rats to see the usefulness of these concentrations in drinking water. Example 1: Inhibition of ferroptosis by iron chelation in TNBS-induced colitis model.

[0112] TNBS colitis is an accepted chemically induced colitis animal model used for testing reagents that affect Chron's disease. A hapten reagent 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis is used in the pre-clinical testing of various chemical or natural compounds in terms of their anti-inflammatory and/or anti-oxidative effects. Inflammatory bowel disease (IBD), consisting of ulcerative colitis (UC) and Crohn's disease (CD), has been shown to involve ferroptosis. Ferroptosis, as a recently recognized form of regulated cell death (RCD), is identified as iron-dependent and caspase-independent nonapoptotic cell death.

[0113] In the TNBS model, H&E (Hematoxylin and Eosin) Staining of the mucosal architecture of the intestines was observed 3 days after induction of colitis. This is a well- accepted model that also covers ferroptosis in diabetes. Iron chelation has been shown to alleviate TNBS-induced colitis via inhibition of ferroptosis (Xu J, et al., Biochemical and Biophysical Research Communications 573 (2021) 48-54).

[0114] Figure 3 shows H&E-stained intestinal mucosa 3 days after induction of colitis with TNBS. CONTROL: mucosal architecture is completed, submucosa no cell infiltration.

TNBS: colon mucosal structured disorder, submucosa cell infiltration. TNBS+ME (ME: MSM + EDTA): colon structure a little bit changed; submucosa a few cells infiltration.

Example 2: Inhibition of ferroptosis by iron chelation in DSS -induced colitis model

[0115] Dextran sulfate sodium (DSS) is a sulfated polysaccharide with variable molecular weights. Administration of DSS causes human ulcerative colitis-like pathologies due to its toxicity to colonic epithelial cells, which results in compromised mucosal barrier function. Iron chelation has been shown to alleviate DSS-induced colitis via inhibition of ferroptosis (Chen Y, et al., Immunology Letters 225 (2020) 9-15).

[0116] Figure 4 shows H&E-stained intestinal mucosa 3 days after induction of colitis with DSS. CONTROL: mucosal architecture is completed, submucosa no cell infiltration. DSS: colon mucosal ulcer (Arrow), goblet cell depletion and structured disorder; submucosal edema, cell infiltration. DSS+ME(MSM+EDTA): colon structure a little bit changed; submucosa a few cells infiltrated. Example 3: Immunohistochemistry (IHC) of iron chelation in DSS-induced colitis model and inhibition of ferroptosis.

[0117] Immunohistochemistry (IHC) by staining with ALDH1, Protein-HNE, Protein- Acrolein, iNOS, one week after the induction of colitis was carried out in the DFSS model of colitis. This is a well-accepted model that also covers ferroptosis in diabetes (Chen 2020). 4- HNE (four hydroxynonenol) and iNOS are accepted as markers of ferroptosis (Yan H-f, et al., Signal Transduction and Targeted Therapy (2021) 6:49)

[0118] Figure 5 illustrates IHC Stained with AEDH1, Protein-HNE, Protein- Acrolein, Protein-MDA in s DSS model of colitis in rats, showing alleviation of microscopic colon damage with MSM+EDTA (ME) treatment. Upper row is Anti- Protein-HNE staining, Lower row is anti-protein-HNE with DAPI. A Normal control. B: DSS induced colitis. C: DSS induced colitis treated with ME.

Example 4: Lens Opacity in Diabetic Rats

[0119] The ability to reduce lens opacity (cataract), activity levels in rats with induced diabetes were observed. Results are shown in the table below. CTRL is Control group with non-diabetic rats dosed with normal water, DT is Diabetic rats’ group with test MSM/EDTA infused drinking water, DC is Diabetic Control rat group dosed with normal (non MSM/EDTA) drinking water. Cataract scale is 0-4 (4 being most acute)

[0120] Statistical significance of the test was established by determining ANOVA p- value.

When the p-value falls below the threshold of p < 0.05, the result of the test is statistically significant.

[0121] Activity levels of diabetic rats +/- administration with MSM and iron chelator were observed and the results are shown below.

Pearson's Chi Sq p=0.0027

Example 5: Effect of iron chelation on chronic inflammation with IL-6 as a marker in rats

[0122] Chronic inflammation with IL-6 as a marker in rats that were dosed ad libitum with normal water or water enriched with MSM and chelator. The MSM/EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal Rats dosed with normal water, NR+ME: Normal Rats + MSM/EDTA drinking water. DR: Diabetic Rats dosed with normal water, DR+ME: Diabetic Rats + MSM/EDTA drinking water.

[0123] Figure 6 illustrates chronic inflammation with IL-6 as a marker in rats that were dosed ad libitum with normal water or water enriched with MSM and chelator. The MSM/EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal Rats dosed with normal water, NR+ME: Normal Rats + MSM/EDTA drinking water. DR: Diabetic Rats dosed with normal water, DR+ME: Diabetic Rats + MSM/EDTA drinking water. A four-fold reduction in IL-6 levels in diabetic rats after administration with MSM and iron chelator.

Example 6: Health of a pancreatic endocrine islet following administration of iron chelators in diabetic rats

[0124] Figure 7A shows a low magnification (lOOx) photomicrograph of a pancreatic lobule, 4pm section of formalin-fixed, paraffin-embedded pancreas, H&E stained. A section of the pancreas (A) from normal rat, shows normal endocrine islets of Langerhans in number and size and normal endocrine acinar tissue. (B) from normal rat dosed orally with M+E also shows normal pancreas islets and acinar tissue. (C) from diabetic rat indicated that pancreatic endocrine islets of Langerhans obviously reduced in number and size. Most of the islets were small, shrunk and inconspicuous. (D) from diabetic rat, dosed orally with M+E had distinctly improved endocrine islets of Langerhans in number and size and there was no shrinkage of acinar tissue. Figure 7B shows a higher magnification photomicrograph (400x) of pancreatic endocrine islets, 4pm sections of formalin-fixed, paraffin-embedded pancreas, H&E stained, (A) An endocrine islet in the pancreas of a normal rat shows interspersed cells in lightly stained exocrine acinar glands, spherical clusters of cells without ducts, and acini. (B). An endocrine islet in pancreas section from normal rat, dosed orally with ME, shows that the histology and morphology were not significantly changed, (C) An endocrine islet in the pancreas of a diabetic rat shows that the islet of Langerhans shrink and become small and inconspicuous (sclerosis of islet, reduction of the cell’s cytoplasm) and show the presence of inter-acinar pancreatitis as evident from leukocytic infiltration in the islets. (D). An endocrine islet of diabetic rat, dosed orally with ME shows that the islet of Langerhans had mild shrinkage and negligible leukocytic infiltration.

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

[0126] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this invention that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claim.

Claims

CLAIMS What is claimed is:

1. A formulation, comprising: a chelating agent or salts thereof, wherein the chelating agent is suitable for long-term consumption; a permeation enhancer that is methylsulfonylmethane (MSM); one or more inert excipients; and a liquid vehicle or carrier; wherein the chelating agent and the permeation enhancer are present in proportions effective to maintain homeostasis of iron levels in the body when consumed in regular doses, and wherein the percentage of chelator is about 0.0001% to 15% and the percentage of permeation enhancer in the composition is about 0.0001% to 30% by weight, respectively.

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

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

4. The formulation of claim 1, wherein the homeostasis of iron levels modulates ferroptosis in a subject.

5. The formulation of claim 4, wherein the ferroptosis is associated with a pathologic disease or condition in a subject.

6. The formulation of claim 5, wherein the pathologic disease or condition is related to an organ selected from: heart, central nervous system, liver, gastrointestinal organs, lung, kidney and pancreas.

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

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

9. The formulation of claim 4, wherein the ferroptosis is reduced by inhibiting irondependent lipid reactive oxygen species (ROS) accumulation.

10. The formulation of claim 1, wherein the homeostasis of iron levels is maintained by chelating iron associated with heme-containing proteins selected from hemoglobin, myoglobin and neuroglobin.

11. The formulation of claim 1, wherein the homeostasis of iron levels is associated with modulating ferroptosis in human cancer cells.

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

13. The formulation of claim 1, wherein the chelating agent is selected from ethylenediamine tetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), cyclohexanediamine tetraacetic acid (CDTA), hydroxy ethylethylenediamine triacetic acid (HEDTA), diethylenetriamine pentaacetic acid (DTP A), dimercaptopropane sulfonic acid (DMPS), dimercaptosuccinic acid (DMSA), aminotrimethylene phosphonic acid (ArPA), citric acid, acetic acid and acceptable salts thereof, and any combinations thereof.

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

15. The formulation of claim 1, wherein the chelating agent is selected from phosphates, pyrophosphates, tripolyphosphates, and hexametaphosphates.

16. The formulation of claim 1, wherein the chelating agent is a nitrogen-containing chelating agents containing two or more chelating nitrogen atoms within an imino group or in an aromatic ring, diimines, or 2,2'-bipyridines.

17. The formulation of claim 1, wherein the chelating agent is a polyamine selected from cyclam (1,4,7,11-tetraazacyclotetradecane), N-(CI-C3O alkyl)-substituted cyclams (e.g., hexadecyclam, tetramethylhexadecylcyclam), diethylenetriamine (DETA), spermine, diethylnorspermine (DENSPM), diethylhomo- spermine (DEHOP), deferoxamine (N'-{5- [Acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4- oxobutanoyl } amino)pentyl]-N-hydroxysuccinamide, or N'- [5-(Acetyl-hydroxy- amino)pentyl]-N-[5-[3-(5-aminopentyl-hydroxy-carbamoyl) propanoylamino]pentyl]-N- hydroxy-butane diamide), desferrioxamine B, desferoxamine B, DFO-B, DFOA, DFB, desferal, deferiprone, pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), ethane-l,2-bis(N-l-amino-3-ethylbutyl-3-thiol).

18. The formulation of claim 1, wherein the chelating agent is a EDTA-4-aminoquinoline conjugate selected from ([2-(Bis-ethoxycarbonyhnethyl-amino)-ethyl]-{[2-(7-chloro- quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([2-(Bis- ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]- methyl }-amino)-acetic acid ethyl ester, ([3-(Bis-ethoxycarbonylmethyl-amino)-propyl]-{[2- (7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([4- (Bis-ethoxycarbonylmethyl-amino)-butyl]-{[2-(7-chloro-quinolin-4-ylamino)- ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([2-(Bis-ethoxymethyl-amino)- ethyl]-{[2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([2-(Bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinolin-4-ylamino)- ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([3-(Bis-ethoxymethyl-amino)- propyl]-{ [2-(7-chloro-quinolin-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester, ([4-(Bis-ethoxymethyl-amino)-butyl]-{[2-(7-chloro-quinolin-4-ylamino)- ethylcarbamoyl]-methyl}-amino)-acetic acid ethyl ester.

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

20. The formulation of claim 1, wherein the chelating agent is poly-asparatic acid or a salt thereof.

21. The formulation of claim 1, wherein the chelating agent is a tetra sodium salt of L- glutamic acid N,N-diacetic acid.

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

23. The formulation of claim 1, further comprising an antibiotic agent.

24. The formulation of 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 formulation of claim 1, wherein the formulation further comprises an emulsifier, wherein the emulsifier is selected from the group consisting of gum arabic, modified starch, pectin, xanthan gum, gum ghatti, gum tragacanth, fenugreek gum, mesquite gum, monoglycerides and di-glycerides of long chain fatty acids, sucrose monoesters, sorbitan esters, polyethoxylated glycerols, stearic acid, palmitic acid, mono-glycerides, di-glycerides, propylene glycol esters, lecithin, lactylated mono- and di-glycerides, propylene glycol monoesters, polyglycerol esters, diacetylated tartaric acid esters of mono- and di-glycerides, citric acid esters of monoglycerides, stearoyl-2-lactylates, polysorbates, 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, furcellaran, Gellan gum, psyllium, curdlan, konjac mannan, agar, and cellulose derivatives, or combinations thereof.

26. The formulation of claim 1, wherein the formulation further comprises an antiinflammatory agent which is a non-steroidal anti-inflammatory (NSAID) drug selected from the group consisting of aceclofenac, aspirin, celecoxib, clonixin, dexibup6fen, 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 of claim 1, wherein the ready-to drink formulation further comprises a flavoring agent is selected from the group consisting of vanilla, vanillin, ethyl vanillin, orange oil, peppermint oil, strawberry, raspberry, synthetic and natural flavors and mixtures thereof.

28. The formulation of claim 1, wherein the chelator is at a concentration of 0.002% to 0.1% w/v.

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

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

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

32. The formulation of claim 30, wherein the ready-to-drink beverage has a pH between 7.0 and 7.4.

33. The formulation of claim 30, wherein the ready-to-drink beverage is selected from the group consisting of noncarbonated beverage, a carbonated beverage, a cola, a root beer, a fruit-flavored beverage, a citrus -flavored beverage, a fruit juice, a fruit-containing beverage7, a vegetable juice, a vegetable containing beverage, a tea, a coffee, a dairy beverage, a protein containing beverage, a shake, a sports drink, an energy drink, and a flavored water.

34. The formulation of claim 30, wherein the formulation is a liquid concentrate, a dissolvable solid, an effervescent tablet, a pill or any form that can be readily reconstituted into a ready-to-drink beverage.

35. The formulation of claim 30, further comprising isotonic agents selected from sugars, buffers and sodium chloride.

36. The formulation of claim 1, wherein the formulation is in a liquid form capable of being administered by a route selected from the group consisting of oral, intranasal, inhalation, intravenous, intra-muscular, transdermal, topical, rectal, vaginal, buccal, injection, sublingual, or combination thereof.

37. The formulation of claim 36, wherein the formulation is a concentrate I the form of a liquid, a dissolvable solid, an effervescent tablet, a pill or a form that can be readily reconstituted.

38. The formulation of claim 36, wherein the formulation is a hydration drip liquid further comprising electrolytes.

39. A method for inhibiting the replication of an RNA virus in a subject, the method comprising providing the formulation according to any of claims 1-38 for administration at a frequency and for a period sufficient to reduce iron ion level in the body below an amount required for reducing hyper-ferritinemia.

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

41. A method for treating or alleviating a ferroptotic condition in a subject, the method comprising providing the formulation beverage according to any of claims 1-38 at a frequency and for a period sufficient to reduce iron ion level in the body below an amount required for ferroptosis at a pathogenic level.

42. The method of claim 41, wherein the ferroptosis is associated with a pathologic disease or condition in a subject

43. The method of claim 42, wherein the pathologic disease or condition is related to an organ in human or animal subject selected from: heart, central nervous system, liver, gastrointestinal organs, lung, kidney and pancreas.

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

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

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

47. The method of claim 41, wherein he ferroptosis is associated with human cancer cells.

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

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

50. The method of claim 49, wherein the heme-containing protein is in a cell selected from red blood cell, muscle cell or neural cell.