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 of related applications) This application claims priority to U.S. Provisional Patent Application No. 63 / 305,099, filed on 31 January 2022, entitled “Novel Liquid Formulation for Iron Chelation,” the contents of which, with due attribution, constitute a part of this specification.

[0002] (Field of invention) The present invention relates to the field of drugs for treating ferroptotic cell death in general, in cases of cancer, aging, viral infection, and other ferroptosis-related pathological conditions. More specifically, it relates to ingestible formulations comprising chelating agents and transport promoters suitable for chelating intracellular iron. In particular, the present invention relates to ready-to-drink beverage formulations comprising transport promoters and chelating agents. In one exemplary embodiment, the present invention relates to a drinkable liquid composition comprising MSM as a transport promoter and a generally safe and acceptable ("GRAS") chelating agent. [Background technology]

[0003] Iron (Fe) plays a major role in human disease states. Certain viruses (RNA viruses) that can cause lethal infections in humans depend on iron for their replication in the human body. For example, treatment with the iron chelator deferipron has been shown to extend the survival of patients with acquired immunodeficiency syndrome (AIDS). Iron overload has been shown to cause ferroptosis, which is part of several human diseases. Ferroptosis is also involved in the debilitating aspects of the aging process. Therefore, limiting iron, whether through oral or intravenous administration of iron chelators or by manipulating key iron regulators, is a promising adjuvant strategy in treating viral infections.

[0004] Coronaviruses are a family of enveloped, single-stranded RNA viruses. In recent decades, two highly pathogenic strains of coronaviruses, 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, sometimes fatal, respiratory illnesses. In December 2019, a new strain of coronavirus (SARS-CoV-2) caused Coronavirus Disease 2019, or Covid-19, which was declared a pandemic by the WHO on March 11, 2020. Common signs of Covid-19 infection include respiratory symptoms, fever, cough, shortness of breath, and difficulty breathing. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, renal failure, and death. As of January 2022, more than 373 million infections and 5.66 million deaths had been reported worldwide.

[0005] While rapid, large-scale vaccination and masking protocols implemented worldwide may have reduced disease severity in many patients, 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 capable of evading immunity produced by vaccines or infection with previous variants. However, currently, no specific treatment exists, and patients rely on general and supportive care, including oxygen supply and broad-spectrum antiviral agents. Remdesivir is a novel nucleotide analog prodrug under development to treat Ebola virus and Middle East Respiratory Syndrome (MERS) disease and has been reported to alleviate pneumonia symptoms of COVID-19 infection. Given the global nature of the pandemic, there is an urgent need for drugs and methods to suppress or block the replication of the SARS-CoV-2 virus, and for such drugs and methods to be widely and readily available to the majority of the world's population susceptible to the SARS-CoV-2 virus and its variants.

[0006] Iron (Fe) has been shown to play two major roles in the severity of Covid-19 disease and its symptoms. Firstly, all coronaviruses (including SARS-CoV-2) depend on 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 evolved to be highly dependent on iron for their replication, and under low iron conditions, replication slows down (Menshawey R, et al. Egyptian Journal of Medical Human Genetics (2020) 21:75). Increased intracellular iron efflux due to increased expression of the iron excretor ferroportin also exhibits antiviral effects against human immunodeficiency virus (HIV) (Liu 2020). Secondly, Covid-19 patients often suffer from iron overload, resulting in ferroptosis, which 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] Hyperferritinemia and altered iron homeostasis are involved in the pathogenesis of COVID-19. Hyperferritinemia has been described as a key feature that predicts the increased risk of death from Covid-19 with high significance (Mehta P, et al., Lancet 2020;395 (March (10229)):1033-4). These studies have shown that serum ferritin levels in COVID-19 non-survivors are twice as high as those in survivors.

[0008] Ferroptosis is a type of controlled necrosis induced by a combination of iron toxicity, lipid peroxidation, and plasma membrane damage. It is a programmed cell death pathway that is dependent on intracellular iron but not on other metals. The non-apoptotic form of cell death may promote the selective elimination of some tumor cells or may be activated in certain pathological conditions. The oncogenic RAS-selective lethal small molecule elastin has been found to induce a specific 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 cell death pattern characterized by the accumulation of lipid reactive oxygen species (ROS). It is primarily caused by an increase in 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 effluent contents form innate and adaptive immunity in health and disease. Excessive or insufficient ferroptotic cell death, coupled with dysregulation of the immune response, is increasingly involved in a wide range of physiological and pathophysiological processes.

[0010] In senile and degenerative diseases, iron levels in the brain inevitably rise. Oxidative stress from excess iron is associated with carcinogenesis. Acute kidney injury (AKI), formerly known as acute renal failure (ARF), is a common and serious disease caused by multiple factors, including ischemia, nephrotoxic drugs, and urinary tract obstruction. While various molecular mechanisms have been proposed to induce or exacerbate AKI, ROS-induced renal injury is considered one of the key mediators. AKI occurs in approximately 5% of hospitalized patients and 30% of critically ill patients, with high morbidity and mortality rates. Furthermore, studies have shown that AKI increases the potential risk of chronic kidney disease and end-stage renal disease in patients. Apart from blood purification, few therapies have made significant progress in preventing AKI. Therefore, new targets or better regimens are still urgently needed to prevent AKI and to facilitate adaptive repair after its onset. Multiple studies suggest that ferroptosis is a promising therapeutic target, particularly in diseases characterized by renal tubular necrosis (Linkermann A. et al. Journal of the American Society of Nephrology. 2014;25(12):2689-2701).

[0011] Iron chelation is widely recognized as improving and / or preventing ferroptosis. Unfortunately, the only chelating agents suggested in the literature are deferoxamine (DESFERAL®), which is approved only for intravenous and subcutaneous administration; 2,2'-bipyridyl (2,2'-dipyridine), a highly toxic substance that has shown efficacy in cell cultures; and cyclopirox olamine (LOPROX®), a chelating fungicide for topical use but not approved for systemic use. All of these have safety concerns and are not suitable for long-term use to prevent ongoing ferroptosis, nor have they been evaluated at non-toxic doses for long-term use. Ferroptosis is one of the promising therapeutic targets, particularly in diseases where renal tubular necrosis is predominant, such as ischemic, cisplatin-toxic, and rhabdomyolysis-induced AKI.

[0012] Generally, lipid peroxidation inhibitors, such as lysyl oxidase inhibitors, ferrostatin-1, and lyproxitin-1, are used to suppress ferroptosis. In some of these models, iron chelators were investigated long before ferroptosis was detected. However, these compounds, such as deferoxamine, were never incorporated into clinical routines despite showing considerable efficacy in ex vivo experiments using renal tubules. Antioxidants and iron chelators (e.g., vitamin E and deferoxamine) have also been observed to suppress ferroptosis by reducing iron availability, but this was only in in vitro experiments and rodent models. Chronic administration of deferoxamine has been reported to have several adverse effects, including acute respiratory distress syndrome, visual impairment, and enhanced Yersinia enterocolitica infection.

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

[0014] Iron chelators need to be administered systemically to subjects, humans, or animals to mitigate and treat diseases, conditions (including aging), and viral infections that can potentially cause ferroptosis. Other iron chelators commonly used in food include EDTA disodium, EDTA calcium disodium, and metaphosphate. These may be available for oral administration, but are generally not absorbed into the body by oral delivery.

[0015] Therefore, there is a need for novel formulations comprising a systemically administered iron chelator and one or more penetrating agents to facilitate absorption of the chelator into the body. In particular, during pandemics caused by rapidly mutating RNA viruses, the desirable formulation for suppressing viral replication is one that can be easily used to lower iron levels in the subject, thereby suppressing viral replication and viral load. [Overview of the project]

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

[0017] Formulations comprising a transport promoter (e.g., MSM) and a chelating agent (e.g., EDTA) for topical application to the eyes, teeth, and skin surfaces have been previously disclosed and patented by the inventors (International Publication 2013 / 166459 by Bhushan et al.; International Publication 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 or a salt thereof, wherein the chelating agent is suitable for long-term intake; an osmotic enhancer, which is methylsulfonylmethane (MSM); one or more inert additives; and a liquid vehicle or carrier, wherein the chelating agent and the osmotic enhancer are present in proportions effective for maintaining homeostasis of iron levels in the body when taken in normal doses, and the percentage of the chelating agent in the composition is about 0.0001% to 15% by weight, and the percentage of the osmotic enhancer is about 0.0001% to 30% by weight.

[0019] In one aspect of the present invention, the formulation is in a liquid concentrated form, a soluble solid form, a foaming tablet form, a pill form, 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 containing electrolytes, vitamins, or other nutrients.

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

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

[0023] In certain aspects of the present invention, the suppression of 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, the suppression of 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 suppressing 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] The method involves a therapeutically effective amount of chelating agent and the following formula (I) [ka] (I) [In the formula, R 1 and R 2 These are independently substituted C2-C6 alkyl, C1-C6 heteroalkyl, and C6-C 14 Aralquil and C2C 12 Selected from heteroaralkyls, where Q is either S or P. This includes administering an effective amount of a formulation consisting of an effective amount of a transport promoter that has the properties of transport to the target.

[0028] The transport accelerator may be, for example, methylsulfonylmethane (MSM; also known as methylsulfone, dimethylsulfone, and DMSO2).

[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, phosporic acid, pyrophosphate, metaphosphate, malic acid polymer, and their acceptable salts, and any combination thereof.

[0030] This invention provides a method for treating or mitigating Covid-19 by suppressing the replication of SAR-CoV2.

[0031] This paper discloses methods for preventing, treating, or alleviating diseases or conditions associated with high iron levels in cells, hyperferritinemia, and ferroptosis.

[0032] These and other embodiments will become apparent from the following description of preferred embodiments, which are to be considered in conjunction with the following drawings, but the changes and modifications therein may affect the spirit and scope of the novel concepts of this disclosure without departing from them. [Brief explanation of the drawing]

[0033] The following drawings form part of this specification and are included to further illustrate specific aspects of the present disclosure, and the invention can be better understood by referring to one or more of these drawings in conjunction with the detailed description of the specific embodiments described herein.

[0034] A patent or application file must include at least one drawing made in color. A copy of the published patent or patent application containing the color drawing will be provided by the Patent Office upon request and payment of the required fees.

[0035] [Figure 1] Figure 1 shows that ferroptosis plays an important role in multisystem diseases, including neurological disorders, heart diseases, liver diseases, gastrointestinal diseases, lung diseases, kidney diseases, and pancreatic diseases. Figure 1 is quoted from Li J. et al. Cell Death and Disease (2020) 11:88.

[0036] [Figure 2] Figure 2 illustrates the vicious cycle of aging mechanisms, accelerated by underlying chronic disease and ferroptosis. The figure shows ferroptosis as a catalyst for inducing a vicious cycle of accelerated aging and chronic disease progression, accompanied by an imbalance in underlying oxidative and antioxidant defense mechanisms. Figure 1 is quoted from Mazhar M, et al. Cell Death Discovery (2021) 7:149 (obtained on January 28, 2022 from https: / / doi.org / 10.1038 / s41420-021-00553-6).

[0037] [Figure 3]Figure 3 shows HE-stained intestinal mucosa 3 days after colitis induction by TNBS. Control: Mucosal structure is complete, and there is no cell infiltration in the submucosa. TNBS: Abnormal colonic mucosal structure, cell infiltration into the submucosa. TNBS + ME (MSM + EDTA): Colonic structure is slightly altered; slight cell infiltration into the submucosa.

[0038] [Figure 4] Figure 4 shows HE-stained intestinal mucosa 3 days after colitis induction by DSS. Control: Mucosal structure is complete, and there is no cell infiltration in the submucosa. DSS: Colonic mucosal ulcer (arrow), goblet cell depletion and structural abnormalities; submucosal edema, cell infiltration. DSS + ME (MSM + EDTA): Colonic structure is slightly altered; a small number of cells infiltrate the submucosa.

[0039] [Figure 5] Figure 5 shows IHC stained with ALDH1, protein-HNE, protein-acrolein, and protein-MDA in a rat DSS model of colitis, demonstrating the reduction of colon damage on microscopic view with MSM+EDTA (ME) treatment. The top row shows anti-protein-HNE staining, and the bottom row shows anti-protein-HNE including DAPI. A: Normal control. B: DSS-induced colitis. C: DSS-induced colitis treated with ME.

[0040] [Figure 6] Figure 6 shows chronic inflammation mediated by IL-6 as a marker in rats given either plain water or water supplemented with MSM and a chelating agent ad libitum. The MSM / EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal rats given plain water, NR+ME: Normal rats + MSM / EDTA drinking water. DR: Diabetic rats given plain water, DR+ME: Diabetic rats + MSM / EDTA drinking water.

[0041] [Figure 7A]Figure 7A shows low-magnification (100x) micrographs of pancreatic lobules stained with HE from 4 μm sections of pancreas fixed in formalin and embedded in paraffin. (A) Sections of the pancreas from normal rats show normal number and size of Langerhans islets and normal acinar tissue. (B) Sections from normal rats orally administered M+E (MSM+EDTA) also show normal pancreatic islets and acinar tissue. (C) Sections from diabetic rats showed a clear decrease in the number and size of Langerhans islets in the pancreatic endocrine islets. Most of the islets were small, contracted, and inconspicuous. (D) Sections from diabetic rats orally administered M+E showed a clear improvement in the number and size of Langerhans islets in the endocrine islets, and there was no contraction of the acinar tissue.

[0042] [Figure 7B] Figure 7B shows high-magnification (400x) micrographs of pancreatic islets stained with HE from 4 μm sections of pancreas fixed in formalin and embedded in paraffin. (A) Pancreatic islets from a normal rat show lightly stained cells scattered within exocrine acini, tubular spherical cell masses, and acini. (B) Pancreatic islets from a normal rat orally administered ME show no significant histological or morphological changes. (C) Pancreatic islets from a diabetic rat show contraction, shrinkage, and inconspicuousness of the islets of Langerhans (sclerosis of the islets, reduction of cytoplasm in cells), and the presence of interacinary pancreatitis, as is evident from leukocyte infiltration in the islets. (D) Pancreatic islets from a diabetic rat orally administered ME show mild contraction of the islets of Langerhans and negligible leukocyte infiltration. [Modes for carrying out the invention]

[0043] The terms used herein generally have their ordinary meanings in the art within the context of the present invention and in the specific context in which each term is used. Specific terms used to describe the present invention are discussed below or elsewhere in this specification to provide practitioners with further guidance 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 does not affect the scope and meaning of the terms, which are the same in the same context whether highlighted or not. It is understood that the same thing can be said in multiple ways. Therefore, different words and synonyms may be used for one or more of the terms discussed herein, and they do not have any special meaning, whether the terms are detailed or discussed. Synonyms for certain terms are provided. Detailed explanation of one or more synonyms does not preclude the use of other synonyms. The use of examples anywhere in this specification, including examples of any terms described herein, is illustrative only and does not in any way limit the scope and meaning of the present invention or any exemplified terms. Similarly, the present invention is not limited to the various embodiments provided herein.

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

[0045] When referring to pharmaceutical ingredients, the term used, for example, “drug,” is intended to encompass not only specific molecular entities but also their pharmaceutically acceptable analogues, including but not limited to salts, esters, amides, prodrugs, conjugates, active metabolites, and other such derivatives, analogues, and related compounds.

[0046] As used herein, the terms “to treat” and “to treat” refer to the administration of a drug or formulation to a clinically manifested human or animal suffering from an adverse condition, disorder or disease, in order to reduce the severity and / or frequency of the symptoms, to eliminate the symptoms and / or their underlying cause, and / or to promote the improvement or repair of the damage. The terms “to prevent” and “prevention” refer to the administration of a drug or composition to a clinically manifested individual susceptible to a particular adverse condition, disorder or disease, and therefore relate to the prevention of the occurrence of the symptoms and / or their underlying cause. Unless otherwise expressly or implicitly stated herein, when the term “to treat” (or “to treat”) is used without mentioning the possibility of prevention, prevention is intended to be included, just as “methods for treating gingivitis” encompass “methods for preventing gingivitis.”

[0047] The terms "any substituent" or "optionally present," as in "any substituent" or "optionally present additive," mean that the component described thereafter (e.g., substituent or additive) may or may not be present, and therefore this description includes both the presence and absence of that component.

[0048] "Medicinally acceptable" means a substance that is not biologically or otherwise undesirable, for example, that can be incorporated into the formulation of the present invention without causing undesirable biological effects or interacting in a harmful manner with any of the other components of the dosage form formulation. However, when the term "medically acceptable" is used to refer to a pharmaceutical excipient, it means that the excipient meets the necessary standards of toxicity and manufacturing testing and / or is included in the Inactive Ingredient Guide prepared by the U.S. Food and Drug Administration. "Pharmacologically active" (or simply "active"), as in the case of a "pharmacologically active" derivative or analogue, as will be explained in more detail below, refers to a derivative or analogue that has the same type of pharmacological activity as the parent drug. As used herein, the terms "to treat" and "to treat" refer to reducing the severity and / or frequency of symptoms, eliminating symptoms and / or their underlying cause, preventing the occurrence of symptoms and / or their underlying cause, and improving or repairing an undesirable condition or injury. Therefore, for example, “treating” a subject includes the prevention of adverse conditions in susceptible individuals, as well as the treatment of individuals with clinical symptoms by suppressing or reducing such conditions. The term “chelating agent” (or “active substance”) refers to a 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, but is not limited to, pharmaceutically acceptable derivatives of active substances specifically described herein, including salts, esters, amides, prodrugs, active metabolites, isomers, analogs, crystalline forms, hydrates, etc. When the term “chelating agent” is used, or when a particular chelating agent is specifically identified, it should be understood that pharmaceutically acceptable salts, esters, amides, prodrugs, active metabolites, isomers, analogs, etc. of that substance are intended as well as the substance itself.

[0049] The “effective” or “therapeutic” amount of an active substance means an amount of substance that is non-toxic but sufficient to provide a beneficial effect. The amount of an “effective” active substance varies from subject to subject, depending on the individual’s age and overall condition, the specific active substance, etc. Unless otherwise specified, the “therapeutic” amount as used herein is intended to include not only an amount effective for treating a harmful condition, but also an amount effective for preventing and / or improving a harmful condition.

[0050] As will be apparent to those skilled in the art who have read the present invention, each of the individual embodiments described and illustrated herein has individual elements and features that can be readily separated from or combined with features of any of the other embodiments without departing from the scope or spirit of the invention. Any described method may be performed in the order of the events described, or in any other logically possible order.

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

[0052] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art in the field relating to this invention. In case of any conflict, this specification, including the definitions, shall prevail.

[0053] The role of iron in Covid-19 Iron overload is increasingly being suggested as a contributing factor to the pathogenesis of COVID-19. Indeed, several symptoms of COVID-19, such as inflammation, hypercoagulation, hyperferritinemia, and immunodeficiency, are reminiscent of iron overload. While iron is essential for all living cells, free, unbound iron resulting from iron dysregulation and overload is highly reactive and potentially toxic due to its role in the generation of reactive oxygen species (ROS). ROS react with cellular lipids, nucleic acids, and proteins, damaging them and consequently activating either acute or chronic inflammatory processes involved in multiple clinical symptoms.

[0054] COVID-19 manifests in numerous complications as well as physiological and biochemical changes. These include, but are not limited to, acute respiratory distress syndrome (ARDS), high concentrations of pro-inflammatory CD4 T cells and cytotoxic granules CD8 T cells, massive cytokine release (cytokine storm), increased coagulation status, hemoglobin damage, and dysregulation of iron homeostasis, including iron overload (Habib, 2021).

[0055] After a 20-month global circulation, the base lineage of SARS-CoV-2 has been almost completely replaced by derived mutant lineages. These lineages are classified by the WHO as variants of concern (VOCs) or variants of note (VOIs) based on genetic, phenotypic, and epidemiological differences. Despite possessing some proofreading ability (a relatively rare function for RNA viruses), SARS-CoV-2 has accumulated approximately 24-25 substitutions per year. A rigorous quantification of the evolutionary process indicates that the observed success of the mutant viruses is the result of adaptive, non-neutral evolution (Kistler KE et al., preprint available January 28, 2022, from https: / / doi.org / 10.1101 / 2021.09.11.459844).

[0056] Iron plays two roles in the severity of Covid-19 disease and its symptoms: (a) All coronaviruses (including SARS, MERS, and SARS-CoV-2) depend on iron for their replication (Shi 2005, Liu 2020, Habib 2021), and under low iron conditions, replication is slowed (Menshawey, 2020). (b) Covid patients often suffer from iron overload, resulting in ferroptosis, which leads to damage to multiple organs (Chen 2021, Yang 2020, Edeas 2020).

[0057] The body's primary storage of iron is in cells containing hemoglobin, myoglobin, and neurorobin. Iron is an essential element for blood production. Over 70% of the body's iron is found in red blood cells as 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 neurorobin. About 6% of the body's iron is a component of certain proteins essential for respiration and energy metabolism, and a component of enzymes involved in the synthesis of collagen and several neurotransmitters. Iron is also necessary for proper immune function. About 20-25% of the body's iron is stored as ferritin, found in cells and in the circulating blood.

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

[0059] The primary entry point for SARS-CoV-2 into cells is by attachment to angiotensin-converting enzyme 2 (ACE2), which is attached to the outer surface of the cell membrane in cells in the lungs, arteries, heart, kidneys, and intestines. Binding of SARS-CoV and the SARS-CoV-2 spike S1 protein to the enzymatic domain of ACE2 on the cell surface leads to endocytosis and the transfer of both the virus and the enzyme into endosomes located within the cell. Liu et al. suggest that the strong affinity of SARS-CoV-2 for the human ACE2 molecule indicates that a key pathogenic molecular step in COVID-19 is the attack of ACE2-positive cells. Over 80% of ACE2 receptors are expressed in a small population of type II alveolar cells (AT2).

[0060] Iron levels in the body are extremely strictly regulated. Sufficient intracellular iron levels support coronavirus replication, while intracellular iron deficiency weakens the replication process by interfering with viral transcription, translation, assembly, and exocytosis. The body's main iron stores are cells containing heme-containing proteins, such as hemoglobin, myoglobin, and neurolobin. Iron is an essential element for blood production.

[0061] To replicate, coronaviruses require iron, and therefore, infecting iron-containing cells is productive for their replication. However, for the Covid virus to enter a cell, the cell must have an ACE2 receptor. Red blood cells, which contain hemoglobin, do not have an ACE2 receptor on their surface. There may be another pathway on the surface of red blood cells that can interact with the SARS-CoV-2 S1 spike protein. One such possible pathway is through interaction with the red blood cell (RBC) Band3 surface protein (Cosic I, et al., Appl. Sci. 2020, 10, 4053; doi:10.3390 / app10114053). However, this virus infects other iron-rich cells that contain an ACE2 surface receptor, including muscle cells (myoglobin) and nerve cells (neuroglobin).

[0062] Hemoglobin, myoglobin, and neuroglobin lose their ability to bind oxygen, hindering its delivery to major organs, which leads to rapid multi-organ failure. Furthermore, free iron released into circulation can cause iron overload, leading to oxidative damage to the lungs and other organs. Iron overload can also cause inflammation and immune dysfunction. These indicate increased iron uptake and storage in iron-binding proteins. In fact, this idea is supported by the increased circulating ferritin (iron storage molecule in the body) levels in COVID-19 patients. Increased iron load leads to increased blood viscosity accompanied by recurrent and diffuse macrocirculatory thrombosis; this may explain unexpected exacerbations and, in some cases, death.

[0063] The iron dependence of viral replication and the regulation of host iron metabolism by RNA viruses highlight the importance of cellular iron homeostasis in the viral life cycle and suggest the usefulness of iron chelating strategies in treating viral infections. One strategy is to directly deplete iron with chelating agents that have a potent and selective affinity for iron ions. Several iron chelating agents, such as deferoxamine (DFO, DESFERAL®), deferipron (DFP, FERRIPROX®), and deferasirox (ICL670, EXJADE®), are approved for clinical use by the U.S. Food and Drug Administration. Iron chelating agents can bind free iron or remove iron from iron-containing proteins. Deferasirox is a membrane-permeable iron chelating agent and was the first oral medication approved by the U.S. Food and Drug Administration (FDA) for chronic iron overload in the body caused by multiple transfusions. Treatment with higher doses of DFP has been shown to extend survival in AIDS patients after HIV-1 infection. Cyclopirox is Fe 3+ It is a synthetic broad-spectrum antifungal agent that binds to trivalent cations such as . Dexrazoxane is a cyclic derivative of EDTA that readily permeates cell membranes. Baicalein is a flavonoid extracted from Scutellaria baicalensis Georgi and has a free 5,6,7-hydroxyl group that forms a complex with iron in a 1:1 stoichiometric ratio.

[0064] Iron chelating drugs can bind to free iron, but they can also remove iron from iron-containing proteins, meaning that iron chelating may have an anti-ferritin effect. In fact, deferoxamine increases lysosomal degradation of ferritin. Iron chelating may play an important role in the control and treatment of COVID-19, but approved iron chelators are not suitable for long-term use. The most widely used approved iron chelator (deferoxamine, DFO) has the drawback of degrading ferritin (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), potentially worsening symptoms. However, food-grade chelators, particularly EDTA, do not have this effect on ferritin.

[0065] ferroptosis The upstream inducers of ferroptosis can be classified into two categories (biological vs. chemical) and can activate two major pathways (exogenous / transporter vs. endogenous / enzymatic pathways). Ferroptosis lacks the typical morphological features of necrosis, such as cytoplasmic and organelle swelling and cell membrane rupture, as well as the features of conventional apoptosis, such as cell contraction, chromatin condensation, apoptotic body formation, and cytoskeletal collapse. In contrast to autophagy, ferroptosis does not involve the formation of a classical closed bilayer membrane structure (autophagy vacuole). Morphologically, ferroptosis manifests primarily as a clear contraction of mitochondria accompanied by increased membrane density, and a decrease or disappearance of mitochondrial cristae, a process distinct from other modes of cell death.

[0066] Current findings indicate that ferroptosis occurs during various pathophysiological processes in the body, including degenerative diseases of the central nervous system, antiviral immune responses, arteriosclerosis, acute kidney injury, diabetes, and ischemia-reperfusion injury (Oxid Med Cell Longevity. 2019; 2019:8010614; published online October 31, 2019). Ferroptosis is characterized by the accumulation of membrane lipid peroxidation products and the uptake of plasma membrane polyunsaturated fatty acids. This type of cell death can be induced by certain small molecules such as elastin and RAS-selective lethal 3 (RSL3). Ferroptosis has been reported to be involved in various pathological processes of brain, kidney, liver, and heart diseases (Sheng X., et al. Physical Chemistry Chemical Physics. 2017;19(20):13153-13159).

[0067] Depletion of glutathione (L-glutamyl-L-cysteinylglycine (GSH)) can lead to iron-dependent accumulation of reactive oxygen species (ROS), particularly lipid ROS, which is sufficient to kill cells (Dixon, 2012). Iron metabolism and lipid peroxidation signaling are considered to be central mediators of ferroptosis. Circulating iron exists in the form of ferric iron (Fe3+) bound to transferrin, thereby reducing free iron levels. Excess iron can lead to the production of ROS that mediate ferroptosis. The biological characteristics of ferroptosis are characterized by the aggregation of iron and ROS. When ferroptosis occurs, as observed under an electron microscope, the cell membrane ruptures and blisters form, the cell nucleus lacks chromatin condensation, mitochondria are reduced, mitochondrial size decreases, bilayer membrane density increases, mitochondrial cristae decrease or disappear, and the outer mitochondrial membrane ruptures.

[0068] Ferroptosis differs from apoptosis and necroptosis, as indicated by its independent mediation in the absence of key effectors of apoptosis and necroptosis, namely BAX, BAK, caspase; mixed-lineage kinase domain-like protein (MLKL), and receptor-interacting serine / threonine kinases (RIPK1 and RIPK3). While ferroptosis has cancer-protective effects against unregulated tumor cells, it may be a root cause of the pathogenesis of various diseases and a hidden cause of many unidentified disease mechanisms. Despite rigorous research in the field of ferroptosis, much remains to be explored to fully understand its mechanisms and role in physiological and pathological conditions. The mechanisms of ferroptosis described by various researchers so far can be simply composed of four steps: (i) inactivation of the cysteine / glutathione countertransport system, (ii) depletion of glutathione and GPx4, (iii) overproduction of lipid ROS, and (iv) excessive cellular iron accumulation.

[0069] Ferroptosis spreads paracrinely via signaling pathways that are not yet clearly defined, but may involve the toxic end products of lipid peroxidation, 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA), which are stable and can react with biomolecules, affecting sites far from their origin. Electron-light nuclei have been found by transmission electron microscopy to be a distinctive and prominent feature of ferroptosis. The morphological features induced intracellularly during ferroptosis, unlike apoptosis, primarily affect mitochondria. Dixon et al. observed mitochondrial contraction and dysfunction, indicating that ferroptosis is directly associated with cellular energy changes and deficiencies, and ultimately cell death, leading to cellular energy and mitochondrial dysfunction. The classical profile of ferroptosis is regulated cell death due to iron-dependent lipid peroxidation, which can be mitigated by iron chelators and lipid antioxidants, and the culprits of lipid peroxidation are generally thought to be ROS that react with polyunsaturated fatty acids (PUFAs) in the membrane, inducing lipid peroxidation. Several ROS production pathways have been proposed, but the detailed mechanism of iron-induced ROS remains unclear.

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

[0071] Iron chelation is widely recognized as improving / preventing ferroptosis. However, the only chelating agents suggested in the literature are deferoxamine (DESFERAL®), which is approved for intravenous and subcutaneous administration only; 2,2'-bipyridyl (2,2'-dipyridine), a highly toxic substance that has shown efficacy in cell cultures; and cyclopirox olamine (LOPROX®), a chelating fungicide for topical use that is not approved for systemic use. All of these have safety concerns and are not suitable for long-term use to prevent ongoing ferroptosis, nor have they been evaluated at non-toxic doses for long-term use.

[0072] Several iron chelators, such as EDTA disodium, EDTA calcium disodium, and metaphosphate, are commonly used in food products. While these are generally not absorbed into the body by oral delivery, these compounds can be used orally to chelate iron and prevent or improve ferroptosis.

[0073] Chelating agents: Chelation is the chemical bonding of a metal to a complex in which the metal is part of a ring. The organic ligand is called a chelator or chelating agent, and the chelate is a metal complex. The more ring-closing atoms there are to the metal atom, the more stable the compound becomes. The stability of a chelate is also related to the number of atoms in the chelate ring. Monodentate ligands, such as H2O and NH3, which have one coordinating atom, are readily decomposed by other chemical processes, while polydentate chelators, which provide multiple bonds to the metal ion, provide more stable complexes. Chlorophyll, the green plant pigment, is a chelate consisting of a central magnesium atom bonded to four complex chelating agents (pyrrole rings). Heme is an iron chelate containing an iron(II) ion at the center of a porphyrin. Chelating agents provide a wide range of metal sequestrants for controlling metal ions in aqueous systems. By forming stable, water-soluble complexes with polyvalent metal ions, chelating agents prevent undesirable interactions by blocking the normal reactivity of the metal ion. EDTA (ethylenediaminetetraacetic acid) is a good example of a common chelating agent that contains a nitrogen atom and a short-chain carboxyl group.

[0074] For the purposes of the present invention, substances that are safe to ingest are suitable for formulating compositions that can be consumed by subjects requiring iron chelation. Examples of iron and calcium chelating agents include, but are not limited to, diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), 1,3-propylenediaminetetraacetic acid (PDTA), ethylenediamine disuccinic acid (EDDS), and ethylene glycol tetraacetic acid (EGTA). Any suitable chelating agent known in the art that is biologically safe and capable of chelating iron, calcium, or other metals is suitable for the present invention.

[0075] The concentration of the chelating agent must be sufficient to transport it across the cell membrane to access intracellular iron. Therefore, the concentration of the chelating agent is determined by the thickness of the cell membrane in 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 (equivalent to 260 ppm (0.0026%) in humans). In some embodiments, the transport enhancer may be present in the formulation of the present invention in amounts ranging from about 0.0001% to about 15% by weight, typically from about 0.001% to about 1% by weight, and more typically from about 0.10% to about 5% by weight.

[0076] Compounds useful as chelating agents in this specification include compounds that coordinate to or form complexes with divalent or polyvalent metal cations and thus function as metal encapsulants for such cations. Accordingly, the term “chelating agent” as used herein includes not only divalent and polyvalent ligands (typically referred to as “chelating agents”) but also monovalent ligands that can coordinate to or form complexes with metal cations.

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

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

[0079] EDTA is widely used as a chelating agent for metals in biological tissues and blood, and its inclusion in various formulations has been suggested. For example, U.S. Patent No. 6,348,508 by Denick Jr. et al. describes EDTA as a sequestering agent for binding 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 by Castillo et al. U.S. Patent No. 6,265,444 by Bowman et al. discloses the use of EDTA as a preservative and stabilizer. However, due to its low permeability across biological membranes, EDTA is generally not applied topically in formulations of significant concentrations.

[0080] Among the chelating / metal-encapsulating substances that may be included in the composition, biocompatible chelating agents may be, but are not limited to, monomeric polyacids, such as 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 any combination thereof.

[0081] Other exemplary chelating agents include phosphates, e.g., pyrophosphates, tripolyphosphates, and hexametaphosphates; chelated antibiotics, e.g., chloroquine and tetracycline; nitrogen-containing chelating agents containing two or more chelated nitrogen atoms in the imino group or aromatic ring (e.g., diimine, 2,2'-bipyridine, etc.); and polyamines, e.g., cyclam(1,4,7,11-tetraazacyclotetradecane), N-(C1-C 30 Alkyl)-substituted cyclamates (e.g., hexadecyclam, 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-(acetyl-hydroxy- Examples include [mino)pentyl]-N-[5-[3-(5-aminopentyl-hydroxy-carbamoyl)propanoylamino]pentyl]-N-hydroxy-butanediamide); also known as desferrioxamine B, desferoxamine B, DFO-B, DFOA, DFB, and desferal), deferipron, pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), and ethane-1,2-bis(N-1-amino-3-ethylbutyl-3-thiol).

[0082] Furthermore, suitable biocompatible chelating agents that may be useful in implementing this disclosure include EDTA-4-aminoquinoline conjugate, for example, Solomon et al., Med. Chem. 2: 133-138. The following are listed in 2006: ([2-(bis-ethoxycarbonylmethyl-amino)-ethyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([2-(bis-ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([3-(bis-ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([4-(bis-ethoxycarbonylmethyl-amino)-butyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino) Examples include (no)-ethyl acetate, ([2-(bis-ethoxymethyl-amino)-ethyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([2-(bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([3-(bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, and ([4-(bis-ethoxymethyl-amino)-butyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate.

[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, etc.

[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 (PASA - BAYPURE® DS100; LANXESS GMBH, Leverkusen, DE). 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 to the SIH (salicyaldehyde isonicotinoylhydrazone) iron chelating agent, which inhibits iron-catalyzed hydroxyl radical generation.

[0086] Inactivated metal ion encapsulating agents are sometimes referred to herein as “prochelating agents,” but metal ion sequestration may involve encapsulation and complexing processes that go beyond chelation itself. The term “prochelating agent” is similar to the term “prodrug,” in that a prodrug is a drug that is therapeutically inactive until it is activated in vivo, and similarly, prochelating agents cannot encapsulate metal ions until they are activated in vivo.

[0087] Transport accelerators: The transport promoter is selected to promote the transport of the chelating agent through the body's tissues, extracellular matrix, and / or cell membranes. An "effective amount" of the transport promoter is an amount and concentration within the formulation of the present invention sufficient to provide a measurable increase in the penetration of the chelating agent through one or more sites of the oral cavity or teeth in a subject compared to the case where the formulation does not contain the transport promoter.

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

[0089] The transport promoter generally has the formula (I)

Chemical formula

[0090] The terms “having a formula” or “having a structure” are not intended to be limiting and are used in the same general sense as the term “containing.” With respect to the structures described above, 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, and cyclohexyl. Unless otherwise specified, the term “alkyl” includes unsubstituted and substituted alkyl groups, where the substituent may be, for example, halo, hydroxyl, sulfhydryl, alkoxy, or acyl. The term “alkoxy” refers to an alkyl group bonded via a single terminal ether bond; 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 condensed together, directly bonded, or indirectly bonded (so that 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 contain one aromatic ring or two fused or bonded aromatic rings, such as phenyl, naphthyl, biphenyl, diphenyl ether, diphenylamine, and benzophenone. "Aryl" includes unsubstituted and substituted aryl groups, where the substituent may be substituted as described above with respect to the "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 aralkyls, but not limited to, include benzyl, 2-phenyl-ethyl, 3-phenyl-propyl, 4-phenyl-butyl, 5-phenyl-pentyl, 4-phenylcyclohexyl, 4-benzylcyclohexyl, 4-phenylcyclohexylmethyl, and 4-benzylcyclohexylmethyl.The term "acyl" refers to substituents 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 heteroatom-containing alkyl and aralkyl groups, 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] The formulation also contains an effective amount of a transport promoter that facilitates the penetration of the formulation component across cell membranes, tissues, and the extracellular matrix. “Effective amount” of the transport promoter represents a concentration sufficient to provide a measurable increase in the penetration of one or more formulation components across membranes, tissues, and the extracellular matrix, as previously stated. Suitable transport promoters include, for example, methylsulfonylmethane (MSM; also known as methylsulfone), 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, while a transport promoter, is essentially a solvent and is therefore not particularly suitable for the formulations according to the present invention. DMSO acts as an extremely potent solvent and, therefore, as a carrier for its solute. In contrast, MSM acts in a completely different manner by forming hydrogen bonds with selected molecules and altering the charge properties of the target molecule, thereby enabling the target molecule to pass through charged barriers such as biological membranes.

[0092] There are differences in chemical structure 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, methylsulfone, and dimethylsulfone. This colorless solid is characterized by sulfonyl functional groups and is considered relatively chemically inert. MSM has the following structure: [ka] On the other hand, dimethyl sulfoxide (DMSO) is an organosulfur compound with the formula (CH3)2SO. This colorless liquid is a widely used polar aprotic solvent that dissolves both polar and nonpolar compounds and is miscible with a wide range of organic solvents and water. DMSO has the following structure: [ka]

[0093] MSM is an odorless, 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 functions as a multifunctional agent herein, insofar as it not only increases cell membrane permeability but also acts as a "transport enhancer" (TFA) that facilitates the transport of one or more pharmaceutical components into oral tissues. Furthermore, MSM itself can provide pharmacokinetics and function as an anti-inflammatory and analgesic agent. MSM is also a source of organosulfur that acts to improve oxidative metabolism in biological tissues and helps reduce scarring. MSM also possesses unique and beneficial solubilizing properties, exhibiting both hydrophilic and hydrophobic characteristics due to its water solubility as described above, but also 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., hydrogen bonds between the oxygen atom of each S=O group and the hydrogen atom of other molecules, as well as van der Waals bonds, i.e., van der Waals bonds between the methyl group and the nonpolar (e.g., hydrocarbon) moiety of other molecules.

[0094] formulation The compositions of the present invention can be formulated using various means. Techniques for formulation and administration can 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 administration to humans or animals, the formulations must meet sterility, pyrogenicity, general safety, and purity standards comparable 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 for incorporation into formulations that are at least partially aqueous include, but are not limited to, thickeners, isotonic agents, buffers, and preservatives, provided that any such additive does not interact in a detrimental manner with any of the other components of the formulation. It should also be noted that preservatives are generally not always necessary, given the fact that the selected chelating agent itself functions as a preservative.

[0096] The chelating and penetrating agents are dissolved in a solvent selected from the following non-limiting list of solvents: 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, the emulsifier being gum arabic, modified starch, pectin, xanthan gum, ghati 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, lactylated mono- and diglycerides, propylene glycol monoesters, polyglycerol esters, mono- and diglycerides The following are selected from the group consisting of diacetylated tartrate esters, citrate esters of monoglycerides, stearoyl-2-lactic acid, polysorbate, succinyl monoglycerides, acetylated monoglycerides, ethoxylated monoglycerides, quillaja, whey protein isolate, casein, soy protein, vegetable protein, pullulan, sodium alginate, guar gum, locust bean gum, tragacanth gum, tamarind gum, carrageenan, fercellaran, 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 selected from the group consisting of vanilla, vanillin, ethyl vanillin, orange oil, peppermint oil, strawberry, raspberry, and mixtures thereof. The flavoring agent may include other synthetic or natural flavors, or combinations thereof.

[0099] In some embodiments, the formulation further comprises an anti-inflammatory agent which is a non-steroidal anti-inflammatory drug (NSAID) 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, lycopheron, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimeslid, oxaprozin, parecoxib, phenylbutazone, piroxicam, lofecoxib, salsalat, sulindac, tenoxicam, tolfenamic acid, tolmetine, or valdecoxib.

[0100] In some embodiments, the ready-to-drink beverage includes an infusion of tea leaves, coffee beans, or cocoa powder. Any suitable isotonic and buffering agents commonly used in oral formulations may be used, but the pH of the formulation should be 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 non-carbonated beverages, carbonated beverages, cola, root beer, fruit-flavored beverages, citrus-flavored beverages, fruit juices, fruit-containing beverages, vegetable juices, vegetable-containing beverages, black tea, coffee, dairy beverages, protein-containing beverages, shakes, sports drinks, energy drinks, and flavored water.

[0102] In a preferred embodiment, the pharmaceutical formulation is administered in the form of an orally ingestible liquid. However, the formulation may be available in any form that can be readily reconstituted into a liquid concentrate, a soluble solid, an effervescent tablet, or any other form that can be readily reconstituted into a drinkable liquid. The concentrate may be a unit dosage form suitable for a single dose of a precise amount. Suitable pharmaceutical formulations and dosage forms are known to those skilled in the art of pharmaceutical formulation and may be manufactured using conventional methods described in relevant texts and literature, for example, Remington: The Science and Practice of Pharmacy, cited herein.

[0103] Chelating agents may be administered in the form of salts, esters, crystals, hydrates, etc., as needed and to the extent pharmaceutically acceptable. Salts, esters, etc., are known to those skilled in synthetic organic chemistry and can be prepared, for example, using standard procedures described in 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 depends on many factors, varies from subject to subject, and depends on the specific chelating agent, the specific 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” refers to any form of pharmaceutical composition containing an amount of chelating agent and transport promoter sufficient to achieve a therapeutic effect with a single or multiple dose. The frequency of administration that provides the most effective results in an efficient manner without overdose varies depending on the characteristics of the particular active substance, including its pharmacological and physical properties, such as both hydrophilicity and other properties.

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

[0106] Suitable pharmaceutical dosage forms for ingestion include aqueous solutions containing the active ingredient. In all cases, the final dosage form must be a fluid that is stable under manufacturing and storage conditions. The liquid carrier or vehicle may be a solvent or liquid dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.) and suitable mixtures thereof. In many cases, it is preferable to include an isotonic agent, such as sugars, a buffer, or sodium chloride.

[0107] Treatment regimens depend on several easily determinable factors, such as the severity of the condition and the responsiveness of the condition being treated, but typically involve one or more treatments per day. Treatment generally involves the ingestion of one or more drinking liquids containing iron chelators and osmotic enhancers. A course of treatment lasts from a day or a few days to several months, or until a consistent and desirable level of iron ions in the body or a significant reduction in the ferroptotic state is achieved.

[0108] The composition of the present invention may further comprise additional drugs or additives appropriate for the indication. In one embodiment, the pharmaceutical composition further comprises a therapeutically effective amount of at least one antibacterial or antifungal agent. In a more specific embodiment, the antibacterial agent is an antibiotic. [Examples]

[0109] The following examples are provided to the art to explain the complete invention and methods for preparing and using embodiments thereof, and are not intended to limit the scope of what the inventors consider to be their discoveries. While efforts have been made to ensure accuracy with respect to the numerical values ​​used (e.g., quantity, temperature, etc.), some experimental error and deviation should be taken into consideration. Unless otherwise specified, parts are by weight, molecular weight is weight-average molecular weight, temperature is in degrees Celsius, and pressure is atmospheric pressure or near atmospheric pressure.

[0110] To evaluate the efficacy of the formulation according to the present invention, rats were given drinking water containing 26 ppm (0.0026%) of EDTA disodium as ad libitum. When adjusted for humans, this corresponds to 260 ppm (0.026%) as ad libitum. Since ad libitum is not possible in humans, the chelating agent is increased from 500-1000 ppm to 0.5% to obtain equivalent results. For animals that can drink ad libitum, equivalent results can be obtained at 10 ppm or higher, depending on the animal's weight and size.

[0111] The experiment was conducted in rats to examine the usefulness of these concentrations in drinking water.

[0112] Example 1: Suppression of ferroptosis by iron chelation in a TNBS-induced colitis model. TNBS colitis is a recognized chemically induced animal model of colitis used to test reagents that affect Crohn's disease. Hapten reagent 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis is used in preclinical studies regarding the anti-inflammatory and / or antioxidant effects of various chemical or natural compounds. Inflammatory bowel disease (IBD), consisting of ulcerative colitis (UC) and Crohn's disease (CD), has been shown to be associated with ferroptosis. Ferroptosis, as a recently recognized form of regulatory cell death (RCD), is identified as iron-dependent and caspase-independent non-apoptotic cell death.

[0113] In the TNBS model, HE (hematoxylin and eosin) staining of the intestinal mucosal structure was observed 3 days after the induction of colitis. This is a widely accepted model that also includes ferroptosis in diabetes. Iron chelation has been shown to reduce TNBS-induced colitis by suppressing ferroptosis (Xu J, et al., Biochemical and Biophysical Research Communications 573 (2021) 48-54).

[0114] Figure 3 shows HE-stained intestinal mucosa 3 days after colitis induction by TNBS. Control: Mucosal structure is complete, and there is no cell infiltration in the submucosa. TNBS: Abnormal colonic mucosal structure, cell infiltration into the submucosa. TNBS + ME (ME: MSM + EDTA): Colonic structure is slightly altered; slight cell infiltration into the submucosa.

[0115] Example 2: Suppression of ferroptosis by iron chelation in a DSS-induced colitis model. Dextran sulfate sodium (DSS) is a sulfated polysaccharide with a variable molecular weight. Administration of DSS induces human ulcerative colitis-like pathology due to its toxicity to colonic epithelial cells, which leads to a decrease in mucosal barrier function. Iron chelation has been shown to alleviate DSS-induced colitis by suppressing ferroptosis (Chen Y, et al., Immunology Letters 225 (2020) 9-15).

[0116] Figure 4 shows HE-stained intestinal mucosa 3 days after colitis induction by DSS. Control: Mucosal structure is complete, and there is no cell infiltration in the submucosa. DSS: Colonic mucosal ulcer (arrow), goblet cell depletion and structural abnormalities; submucosal edema, cell infiltration. DSS + ME (MSM + EDTA): Colonic structure is slightly altered; a small number of cells infiltrate the submucosa.

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

[0118] Figure 5 shows IHC stained with ALDH1, protein-HNE, protein-acrolein, and protein-MDA in a rat DSS model of colitis, demonstrating the reduction of colon damage on microscopic view with MSM+EDTA (ME) treatment. The top row shows anti-protein-HNE staining, and the bottom row shows anti-protein-HNE including DAPI. A: Normal control. B: DSS-induced colitis. C: DSS-induced colitis treated with ME.

[0119] Example 4: Lens opacity in diabetic rats We observed the ability to reduce lens opacity (cataracts) and activity levels in diabetic-induced rats. The results are shown in the table below. CTRL is the control group of non-diabetic rats given normal water, DT is the group of diabetic rats given drinking water infused with test MSM / EDTA, and DC is the group of diabetic control rats given normal (non-MSM / EDTA) drinking water. The cataract scale ranges from 0 to 4 (4 being the most acute). [Table 1]

[0120] The statistical significance of the study was confirmed by determining the ANOVA p-value. If the p-value is below the threshold of p < 0.05, the results of the study are considered statistically significant. [Table 2]

[0121] The activity levels of diabetic rats administered with and without MSM and iron chelators 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 Chronic inflammation mediated by IL-6 as a marker in rats given either plain water or water supplemented with MSM and a chelating agent ad libitum. The MSM / EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal rats given plain water, NR+ME: Normal rats + MSM / EDTA drinking water. DR: Diabetic rats given plain water, DR+ME: Diabetic rats + MSM / EDTA drinking water.

[0123] Figure 6 shows chronic inflammation markerized by IL-6 in rats given either plain water or water supplemented with MSM and chelating agents ad libitum. The MSM / EDTA concentrations in the water were 26 ppm EDTA and 54 ppm MSM. NR: Normal rats given plain water, NR+ME: Normal rats + MSM / EDTA drinking water. DR: Diabetic rats given plain water, DR+ME: Diabetic rats + MSM / EDTA drinking water. A four-fold decrease in IL-6 levels in diabetic rats after administration of MSM and iron chelating agents.

[0124] Example 6: Health of pancreatic islets after administration of iron chelating agents in diabetic rats Figure 7A shows low-magnification (100x) micrographs of pancreatic lobules stained with HE from 4 μm sections of pancreas fixed in formalin and embedded in paraffin. (A) Sections of the pancreas from normal rats show normal number and size of Langerhans islets, as well as normal acinar tissue. (B) Sections from normal rats orally administered M+E also show normal pancreatic islets and acinar tissue. (C) Sections from diabetic rats showed a clear decrease in the number and size of Langerhans islets in the pancreatic endocrine islets. Most of the islets were small, contracted, and inconspicuous. (D) Sections from diabetic rats orally administered M+E showed a clear improvement in the number and size of Langerhans islets in the endocrine islets, and there was no contraction of the acinar tissue. Figure 7B shows high-magnification (400x) micrographs of pancreatic islets stained with HE from 4 μm sections of pancreas fixed in formalin and embedded in paraffin. (A) Pancreatic islets from a normal rat show lightly stained cells scattered within exocrine acini, tubular spherical cell masses, and acini. (B) Pancreatic islets from a normal rat orally administered ME show no significant histological or morphological changes. (C) Pancreatic islets from a diabetic rat show contraction, shrinkage, and inconspicuousness of the islets of Langerhans (sclerosis of the islets, reduction of cytoplasm in cells), and the presence of interacinary pancreatitis, as is evident from leukocyte infiltration in the islets. (D) Pancreatic islets from a diabetic rat orally administered ME show mild contraction of the islets of Langerhans and negligible leukocyte infiltration.

[0125] All publications and patent applications cited herein are incorporated herein by reference in the same manner as indicated, with each individual publication or patent application being specifically and individually incorporated by reference.

[0126] Although the above invention is described in some detail by examples and illustrations for the purpose of clarifying the understanding, it will be readily apparent to those skilled in the art that certain changes and modifications can be made in light of the teachings of the present invention without departing from the spirit or scope of the appended claims.

Claims

1. Chelating agents suitable for long-term intake, or chelating agents or their salts; Methylsulfonylmethane (MSM) is a penetration enhancer; One or more inert additives; and Liquid vehicle or carrier A formulation containing; Chelating agents and osmotic enhancers are present in proportions that are effective in maintaining homeostasis of iron levels in the body when taken at normal doses, and A formulation wherein the percentage of chelating agents in each composition is approximately 0.0001% to 15% by weight, and the percentage of penetration enhancers is approximately 0.0001% to 30% by weight.

2. The formulation according to claim 1, wherein the iron level homeostasis is sufficient to suppress 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 iron level homeostasis regulates ferroptosis in the subject.

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

6. The formulation according to claim 5, wherein the pathological disease or condition relates to 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 related to aging in the subject.

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

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

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

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

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

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 its 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, EDTA calcium disodium, and combinations thereof.

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

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

17. The chelating agent is cyclam(1,4,7,11-tetraazacyclotetradecane), N-(C 1 -C 30 Alkyl)-substituted cyclamates (e.g., hexadecyclam, 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-(acetyl-hydroxy-amino)pentyl] The formulation according to claim 1, wherein the polyamine is selected from [-N-[5-[3-(5-aminopentyl-hydroxy-carbamoyl)propanoylamino]pentyl]-N-hydroxy-butanediamide), desferrioxamine B, desferoxamine B, DFO-B, DFOA, DFB, desferal, deferipron, pyridoxal isonicotinoyl hydrazone (PIH), salicylaldehyde isonicotinoyl hydrazone (SIH), and ethane-1,2-bis(N-1-amino-3-ethylbutyl-3-thiol).

18. The chelating agents are ([2-(bis-ethoxycarbonylmethyl-amino)-ethyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([2-(bis-ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([3-(bis-ethoxycarbonylmethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([4-(bis-ethoxycarbonylmethyl-amino)-butyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([2-(bis The formulation according to claim 1, which is an EDTA-4-aminoquinoline conjugate selected from (-ethoxymethyl-amino)-ethyl)-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([2-(bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, ([3-(bis-ethoxymethyl-amino)-propyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate, and ([4-(bis-ethoxymethyl-amino)-butyl]-{[2-(7-chloro-quinoline-4-ylamino)-ethylcarbamoyl]-methyl}-amino)-ethyl acetate).

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 formulation according to claim 1, wherein the chelating agent is a natural chelating agent selected from citric acid, phytic acid, lactic acid, acetic acid and its salts, and curcumin.

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

24. The formulation 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 formulation further contains an emulsifier, the emulsifier being gum arabic, modified starch, pectin, xanthan gum, ghati gum, tragacanth gum, fenugreek gum, mesquite gum, monoglycerides and diglycerides of long-chain fatty acids, sucrose monoester, sorbitan ester, polyethoxylated glycerol, stearic acid, palmitic acid, monoglycerides, diglycerides, propylene glycol ester, lecithin, lactylated mono- and di-glycerides, propylene glycol monoester, polyglycerol ester, diacetylated tartaric acid of mono- and diglycerides The formulation according to claim 1, selected from the group consisting of sterol, citrate ester of monoglycerides, stearoyl-2-lactic acid, polysorbate, succinyl monoglycerides, acetylated monoglycerides, ethoxylated monoglycerides, quillaja, whey protein isolate, casein, soy protein, plant protein, pullulan, sodium alginate, guar gum, locust bean gum, tragacanth gum, tamarind gum, carrageenan, fercellan, gellan gum, psyllium, curdlan, konjac mannan, agar, and cellulose derivatives, or combinations thereof.

26. The preparation according to claim 1, further comprising an anti-inflammatory agent which is a nonsteroidal anti-inflammatory drug (NSAID) selected from the group consisting of aceclofenac, aspirin, celecoxib, clonixin, dexibprofen, dexketoprofen, diclofenac, diflunisal, droxicam, etodolac, etoricoxib, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen, indomethacin, isoxicam, ketoprofen, ketorolac, lycopheron, lornoxicam, loxoprofen, lumiracoxib, meclofenamic acid, mefenamic acid, meloxicam, nabumetone, naproxen, nimeslide, oxaprozin, parecoxib, phenylbutazone, piroxicam, lofecoxib, salsalat, sulindac, tenoxicam, tolfenamic acid, tolmetine, or valdecoxib.

27. The preparation according to claim 1, wherein the ready-to-drink preparation further comprises a flavoring agent selected from the group consisting of vanilla, vanillin, ethyl vanillin, orange oil, peppermint oil, strawberry, raspberry, synthetic and natural flavoring agents 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 beverage that can be consumed as is.

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

32. The formulation according to claim 30, wherein the pH of the ready-to-drink beverage is 7.0 to 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, black 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 is in any form that can be readily reconstituted into a liquid concentrate, a soluble solid, an effervescent tablet, a pill, or a ready-to-drink beverage.

35. The formulation according to claim 30, further comprising a sugar, a buffer, and an isotonic agent selected from sodium chloride.

36. The preparation according to claim 1, wherein the preparation is in liquid form and 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 a combination thereof.

37. The formulation according to claim 36, wherein the formulation is in the form of a concentrated liquid, a soluble solid, an effervescent tablet, a pill, or a form that can be easily reconstituted.

38. The formulation according to claim 36, wherein the formulation is a water replacement drop solution further containing electrolytes.

39. A method for suppressing the replication of RNA viruses in a subject, comprising providing a formulation according to any one of claims 1 to 38 for administration at a frequency and duration sufficient to reduce the iron ion level in the body to below the amount necessary to alleviate 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 for treating or mitigating a ferroptotic state in a subject, comprising providing a formulation beverage according to any one of claims 1 to 38 at a frequency and for a period of time sufficient to reduce the iron ion level in the body to below the amount required for pathogenic ferroptosis.

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

43. The method according to claim 42, wherein the pathological disease or condition relates to an organ in 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 related to aging in the subject.

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

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

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

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

49. A method for chelating intracellular iron in a subject, comprising providing a formulation beverage according to any one of claims 1 to 38 at a frequency and duration sufficient to reduce the level of iron ions bound to a heme-containing protein selected from hemoglobin, myoglobin, and neurorobin.

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

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