Compositions and methods for the treatment and prevention of pathogens

Abstract

The present invention provides a novel, storable, and stable disinfectant composition that utilizes a solid precursor of oxidized chlorine. The components in the device are free of stability issues because the solid precursor dissolves instantly in water or a pharmaceutically acceptable diluent, adjuvant, or carrier, and is combined with an activator (e.g., adipic acid or succinic acid or a salt thereof), optionally combined with a thickener, and optionally combined with a dye. The primary product of the present invention is produced by dissolving an effervescent material in tablet or granular form. The resulting solution is a disinfectant useful for treating a wide range of pathogenic bacterial and / or viral, fungal, or parasitic pathogens, including the indicated microorganisms.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention generally relates to compositions comprising a combination of solid precursors of chlorine in one or more oxidation states for the treatment and prevention of pathogens. [Background technology]

[0002] Background of the Invention Microorganisms include bacteria, fungi, archaea, parasites, protozoa, and viruses; they are the oldest known forms of life. These types of microorganisms can be either free-living or parasitic.

[0003] Free-living microorganisms, such as bacteria or fungi, have the potential to grow on or within a host organism, which can infect and cause pathology in the host organism, potentially leading to illness or death.

[0004] Infectious diseases are a leading cause of death worldwide, accounting for over 13 million deaths annually, including nearly two-thirds of all childhood mortality. Furthermore, antibiotic resistance is on the rise, contributing to the morbidity of a wide range of human diseases, including pneumonia, tuberculosis, and cholera. Of particular concern is the number of human pathogens developing multidrug resistance to conventional antibiotics. Introducing new, more potent derivatives of existing antibiotics provides only a temporary solution, as existing resistance mechanisms quickly adapt to accommodate the new derivatives (Hoiby et al., Int. J. Antimicrob. Agents 2010). While resistant Gram-positive bacteria pose a significant threat, the emergence of multidrug-resistant (MDR) strains of common Gram-negative pathogens, such as Escherichia coli, is of particular concern. Pan-resistant or extensively drug-resistant are currently terms commonly used to describe clinically important isolates of Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterobacteriaceae that are resistant to virtually all antibiotics.

[0005] Viruses are also a major concern in infectious disease epidemiology. Serious viral outbreaks, many of zoonotic origin, are becoming increasingly common. For example, the SARS (Severe Acute Respiratory Syndrome) and MERS (Middle East Respiratory Syndrome) outbreaks in the early to mid-2000s, the H1N1 pandemic in 2009, and the subsequent SARS CoV-2 pandemic in 2020 have generated significant attention regarding both the treatment and prevention of the spread of these viral pathogens.

[0006] Many viruses that infect the respiratory tract are transmitted through droplet infection. In this case, respiratory droplets containing the virus are expelled by infected individuals and are picked up by others through direct contact or contact with the surface on which the droplets are attached. Typically, infection progresses through the binding of the virus to receptors on mucous membranes or epithelial cells, and then through invasion into the nose, eyes, ears or mouth. In addition, other viruses are transmitted through aerosol particles containing the virus or are airborne. In either case, the virus can survive for several hours to several days after emergence from infected individuals.

[0007] The infection can be caused by a microorganism, such as a virus, a bacterium, a fungus, a spore, a parasite, or a combination thereof, as described herein. The virus can be any virus, including, but not limited to, adenovirus, human immunodeficiency virus (HIV), rhinovirus, influenza virus (e.g., influenza A), and hepatitis (e.g., hepatitis A). The SARS-CoV virus, identified in 2002 as the cause of the Severe Acute Respiratory Syndrome (SARS) pandemic, and the MERS-CoV virus, identified in 2012 as the cause of Middle East Respiratory Syndrome (MERS), are examples of pathogenic viruses.

[0008] Other problematic microorganisms include, but are not limited to, rotavirus, respiratory syncytial virus, herpes simplex virus, varicella-zoster virus, rubella virus, and other common viruses. Bacterial infections include those caused by, for example, one or more of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, E. coli, Staphylococcus aureus, Bacillus athrophaeus, Streptococcus pyogenes, Samonella choleraesuis, Shingella dysenteriae, and Mycobacterium tuberculosis. Additionally, problematic fungi and yeasts may include, for example, one or more of Candida albicans, Bacillus subtilis, Trichophyton mentagrophytes, and Bacillus athrophaeus.

[0009] Conventional compositions and methods for disinfecting inanimate surfaces or contaminated epithelia are not sufficient to inactivate all of these infectious agents. Current forms of conventional disinfectant compositions and methods may require long, impractical exposure times, or harmful or corrosive solutions or vapors that cannot be used on expensive equipment or living tissue, and therefore cannot provide a fast-acting solution to the increasing health risks from drug- and disinfectant-resistant agents. In particular, it is clear that there is a significant unmet medical need for new methods of treating resistant microorganisms and viruses that are effective against key steps in the invasion of mammalian cells by microorganisms via biological mechanisms, and that are also effective outside of mammalian biology. [Prior art documents] [Non-patent literature]

[0010] [Non-Patent Document 1] Hoiby et al, Int. J. Antimicrob. Agents 2010 Summary of the Invention [Means for solving the problem]

[0011] Summary of the Invention The present invention generally relates to compositions for the elimination of pathogenic viruses, bacteria, and other infectious agents. In a preferred embodiment, the present invention comprises a solid precursor of chlorine in one or more oxidation states, an acid activator and its salt, and a base activator. In some embodiments, the acid activator and the base activator are solid. The compositions of the present invention may also include a thickener and / or a dye or redox-activated dye. The compositions of the present invention are formulated as effervescent powders, tablets, or granules. When dissolved in water, the resulting solution provides a disinfectant for treating a wide range of bacterial, viral, fungal, and parasitic pathogens.

[0012] The compositions of the present invention are provided as effervescent powders, tablets, or granules to be dissolved in water or a pharmaceutically acceptable diluent, adjuvant, or carrier. The oxidized chlorine species is combined with an acid activator and a base activator in the tablet, and the acid activator acts synergistically with the oxidized chlorine. The acid activator may preferably be a carboxylic acid and / or its salt, preferably a carboxylic acid and / or its salt having a melting point above 20 degrees Celsius. If necessary, the tablets described herein may contain a thickener and / or optional dye. The compositions of the present invention are useful as disinfectants for treating a wide range of bacteria, viruses, and / or other pathogens.

[0013] In certain aspects, the present invention provides antimicrobial compositions comprising a solid chlorine oxidizing species salt, an acid activator, a base activator, and a pharmaceutically acceptable diluent, adjuvant, or carrier. The solid chlorine oxidizing species salt is represented by the formula M n+ [Cl(O) x ] n n-where M is an alkali metal, alkaline earth metal, or transition metal ion, n is 1 or 2, and x is an integer between 1 and 4, inclusive. Acid activators are based on the formula RN(COOH) m where R N is a saturated or unsaturated organic moiety containing 1 to 25 carbon atoms, optionally substituted with oxygen or hydrogen to form a functional group that cannot be oxidized by chlorinated species, and m is an integer from 0 to 10, or a salt thereof. The base activator can react with the acid activator to produce carbon dioxide and is represented by the formula (M n+ ) s ((H) t CO3) u where n is 1 or 2, s is 1 or 2, t is 0 or 1, u is 1 or 2, t+s+n is an integer between 1 and 5 inclusive, and M is an alkali metal, alkaline earth metal, or transition metal.

[0014] Preferably, the antimicrobial composition comprises a compound of formula M n+ [Cl(O) x ] a w- wherein M is an alkali metal, alkaline earth metal, or transition metal ion; n, a, and w are each independently 1 or 2; and x is an integer from 1 to 4; and a solid chlorine oxide species salt of the formula: RN(COOH). m wherein R N is a saturated or unsaturated organic moiety containing 1 to 25 carbon atoms optionally substituted with oxygen or hydrogen to form a functional group that cannot be oxidized by chlorinated species, and m is an integer from 0 to 10, or a salt thereof; with a pharmaceutically acceptable diluent, adjuvant, or carrier; and z+ ) s ((H) t CO3) uwherein z is 1 or 2, s is 1 or 2, t is 0 or 1, u is 1 or 2, t+s+z is an integer from 1 to 5, and M′ is an alkali metal, alkaline earth metal, or transition metal, and a base activator capable of reacting with the acid activator to produce carbon dioxide.

[0015] In some embodiments, the chlorine oxide salt comprises an alkali metal salt or alkaline earth metal salt of hypochlorous acid. In other embodiments, the chlorine oxide salt comprises an alkali metal salt or alkaline earth metal salt of chlorite. In certain embodiments, the acid activator is selected from the group consisting of monobasic acids, dibasic acids, tribasic acids, dibasic acids, propionic acid, lactic acid, succinic acid, glutaric acid, pyruvic acid, citric acid, malic acid, oxaloacetic acid, tartaric acid, adipic acid, fumaric acid, heptanedioic acid, octanedioic acid, dibasic acids containing at least 7 carbon atoms, and derivatives thereof.

[0016] In some embodiments, the base activator is selected from sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate.

[0017] In some embodiments, the antimicrobial compositions of the present invention further comprise a water-soluble polyol binder, hi some embodiments, the water-soluble polyol binder is selected from the group comprising monosaccharides, disaccharides, polysaccharides, or monohydroxy or polyhydroxy acids.

[0018] The compositions of the present invention may further comprise a lubricant. Preferred lubricants include dextrose, lactose, sorbitol, ascorbic acid, sorbitol, mannitol, sodium benzoate, potassium sorbate, polyethylene glycol and its derivatives, succinic acid, adipic acid, glutaric acid, and clavulanic acid. In some embodiments, the compositions of the present invention are formulated into an aqueous solution, gel, cream, ointment, or oil. In some embodiments, the dissolved antimicrobial composition has an osmolality ranging from about 0.1 mOsm to about 500 mOsm. In some embodiments, the dissolved composition has a pH between pH 4 and pH 8.

[0019] The compositions of the present invention may further comprise a viscosity enhancing agent. In some embodiments, the viscosity enhancing agent comprises a water-soluble gelling agent. In other embodiments, the water-soluble gelling agent is selected from the group consisting of polyacrylic acid, polyethylene glycol, poly(acrylic acid)-acrylamidoalkylpropane sulfonic acid copolymer, phosphinopolycarboxylic acid, poly(acrylic acid)-acrylamidoalkylpropane, and sulfonic acid-sulfonated styrene terpolymer.

[0020] The compositions of the present invention may further comprise a dye comprising a color that provides a visual indication of the presence of an oxidized chlorine compound. In some embodiments, the dye comprises a redox dye. In some embodiments, the color and color intensity of the dye depend on the oxidation state of the oxidized chlorine compound.

[0021] In some embodiments, the solid components of the antimicrobial composition are contained within an effervescent powder, granules, or tablet.

[0022] In certain aspects, the present invention provides antimicrobial compositions useful in antiviral and antibacterial applications.

[0023] The formulations of the present invention are useful in inhalation therapy using asthma inhalers, nebulizers, or sprayers to combat viral infections of the upper respiratory tract in mammals.

[0024] The formulations of the present invention are useful for preventing or healing skin or wounds.

[0025] The formulations of the present invention are useful in the treatment of mastitis or any other infectious disease in animals or agricultural breeding.

[0026] The formulations of the present invention are useful in the treatment of any infectious disease in aquatic animal breeding.

[0027] The formulations of the present invention are useful in the treatment of any chemical or biological warfare agent.

[0028] In certain aspects, the present invention provides methods for preparing effervescent powders, granules, and tablets derived from the antimicrobial compositions. In some embodiments, the powders, granules, and tablets derived from the antimicrobial compositions are prepared using wet granulation, a fluid bed dryer, or a vacuum granulator.

[0029] In some embodiments, the present invention provides a method for neutralizing chemical or biological weapons using dissolving an effervescent material in a liquid phase and producing a vapor or aerosol in a target area or territory. [Brief explanation of the drawings]

[0030] [Figure 1] Figure 1 is a schematic diagram illustrating an exemplary effervescent tablet for dispensing a disinfecting composition according to an embodiment of the present invention. Figure 1 shows an effervescent tablet prepared from an effervescent powder or granules containing: 1. a chlorine oxide species or salt thereof; 2. an acid activator containing at least one carboxylic acid functionality; 3. an optional salt of a molecule containing at least one carboxylic acid functionality; 4. a base activator; 5. an optional binder; 6. an optional lubricant; 7. an optional thickener; 8. an optional dye; and 9. optional sodium chloride to achieve an osmolality close to that of blood. Tablets can be prepared from the effervescent powder or granule-derived materials of the present invention using wet granulation, a fluidized bed dryer, a vacuum granulator, or any other method suitable for tablet manufacturing. DETAILED DESCRIPTION OF THE INVENTION

[0031] Detailed Description of the Invention The present invention generally relates to compositions comprising a combination of solid and liquid precursors of oxidized chlorine and an activator (e.g., glutaric acid or its salts), and one or more additional ingredients. The use of such compositions acts as antimicrobial and disinfectant agents for the treatment of a wide range of bacterial and / or viral pathogens on a variety of biological and non-biological surfaces and environments.

[0032] The compositions of the present invention can be formulated as solids, for example, in effervescent powders, tablets, or granules. This addresses the problem of reduced shelf life observed in solutions of hypochlorous acid or chlorine dioxide described in the prior art. More specifically, the immediate generation of ready-to-use formulations of oxidized chlorine species from solid precursor API-P can be carried out in effervescent tablets at the point of use. The effervescent powders, tablets, or granules of the present invention are used for distribution, but exhibit long-term stable storage of solid compositions consistent with the present invention. In one example, the solid precursor of oxidized chlorine, glutaric acid or its salt, thickener, and pigment, are then combined to prepare the composition at the desired time and on-site.

[0033] By way of background, chlorine oxides or chlorine oxides (also referred to herein as "OC") constitute a large class of chemical species, commonly found in nature as well as in biological systems in mammals. Chlorine oxides can also exist as neutral compounds or as ions, so-called oxyanions. Several oxyanions exist for chlorine, including the corresponding hypochlorite anion (ClO). - ), chlorite anion (ClO2 - ), chlorate anion (ClO3 - ) or perchlorate anion (ClO4 - It can have oxidation states of +1, +3, +5, or +7, depending on the pH. The standard reduction potential of hypochlorous acid (HOCl) at low pH is +1.49, and for chlorous acid (HClO2), the standard reduction potential is 1.64, but at basic pH it is +0.89 and +0.78, respectively. At pHs between 5 and 7, the reduction potential is greater than +1.

[0034] As a result, hypochlorite and chlorite are generally the most useful oxidation states with the potential to kill microorganisms and parasites at low pH. In particular, chloride ion Cl - is in its most stable oxidation state and is not reactive or effective as a disinfectant. Chlorate and perchlorate salts in oxidation states +5 and +7 are more reactive than the lower oxidation states and can be more difficult to handle.

[0035] The hypochlorite ion has the chemical formula ClO - where chlorine (Cl) is in the oxidation state +1, and the lower-energy oxidation state of Cl is -1, making this a potentially unstable oxidation state. Both hypochlorite and chlorite ions combine with several cations to form hypochlorite and chlorite as salts of these chlorine oxides. Common examples include sodium hypochlorite (household bleach) and calcium hypochlorite, which are the primary active ingredients in commercial products containing bleach powder, chlorine powder, or chlorinated lime, commonly used for water treatment (e.g., swimming pools, etc.). Chlorite and hypochlorite ions, also referred to herein as "primary chlorine oxides," are useful in a variety of situations. Sodium chlorite and sodium hypochlorite are powerful oxidizing agents used in water purification, disinfection, and bleaching and deodorizing animal products.

[0036] Because sodium hypochlorite produces highly toxic chlorine gas under acidic conditions, aqueous solutions commercially available for home use are strongly basic, with the pH adjusted using sodium hydroxide.

[0037] Chlorite and hypochlorite are also useful for treating various diseases or conditions. For example, chlorite and hypochlorite are useful for treating infectious diseases, as described in U.S. Patent Nos. 4,725,437 and 4,851,222. Chlorite and hypochlorite can also be used to treat HIV, recurrent prostate cancer, cystitis, and chronic active hepatitis C disease (see, for example, McGrath et al., Development of WFlO, a novel macrophage-regulating agent, Curr. Opin. Investig. Drugs, 3(3):365-73 (March 2002)). Chlorite and hypochlorite have also been described for use in treating oral or periodontal diseases or conditions, such as gingivitis (see, for example, U.S. Patent No. 6,350,438).

[0038] Hypochlorous acid (HOCl) is widely used as a broad-spectrum household and industrial disinfectant and deodorizer. It is a weak acid known to rapidly inactivate bacteria, algae, fungi, and other organisms, making it an effective agent across a wide range of microorganisms. Furthermore, because hypochlorous acid is a weak acid and people naturally produce certain compounds that allow them to tolerate hypochlorous acid, hypochlorous acid is generally non-toxic to humans. Due to its biocidal properties combined with its safety profile, hypochlorous acid has emerged as having many beneficial applications across many different industries, including healthcare, food service, food retail, agriculture, wound care, laboratories, hospitality, dentistry, and floriculture.

[0039] When chlorine dissolves in water, hypochlorous acid is formed. Specifically, acidification of hypochlorite produces hypochlorous acid, where the chlorine atom is in the oxidation state +1. Hypochlorous acid is Cl - In the presence of HCl, HCl exists in equilibrium with chlorine gas, which can escape from solution. The equilibrium is pH dependent, as shown in the following equation (Equation 1): [ka]

[0040] Referring to the equation above (Equation 1), a high pH drives the reaction to the right, favoring the disproportionation of chlorine to chloride and hypochlorite, while a low pH drives the reaction to the left, favoring the release of chlorine gas (Cl2).

[0041] Chlorine gas (Cl2) is a toxic and powerful oxidizing agent. One undesirable property of chlorine gas is its tendency to insert Cl into hydrocarbons in radical reactions to form monohaloalkanes or polyhaloalkanes and other chlorinated hydrocarbons (see Barhorst and Kubiak, Environ. Sci. Pollut. Res. (2009) 16, pp. 582–589). This is potentially dangerous because halogenated hydrocarbons are considered pollutants and potentially carcinogenic. Therefore, chlorine gas is not a desirable compound for use at higher concentrations in natural or medical applications. At lower concentrations, chlorine contributes to the powerful disinfecting properties of hypochlorous acid in equilibrium, and the risk of medical side effects is reasonably low. However, as long as the pH of the hypochlorous acid solution is controlled and maintained between 4 and 8, the amount of Cl2 is negligible.

[0042] The antibacterial effect of hypochlorous acid is highly pH-dependent, providing the highest level of activity at pH values ​​between 4 and 5.5, but is also highly active at physiological pH (7.1 to 7.5). This means that hypochlorite solutions should ideally be buffered with a weak organic acid and its corresponding metal salt to have their maximum antibacterial activity.

[0043] In general, formulations containing chlorine oxide are effective antimicrobial agents, possessing antibacterial and antiprotozoal properties that have proven useful in disinfection technologies for human and animal health. However, prior art formulations have drawbacks. For example, HOCI, a weak acid, is unstable and impure when prepared under conventional conditions. As a result, there is a need for a more controlled, instantaneous preparation method that can provide an on-site supply of chlorine oxide with stability that allows for its intended short-term use.

[0044] While the alkalinity of household hypochlorite bleach can be approximately pH 12, solutions of HOCl are weakly acidic, with a pKa value of approximately 7. Therefore, HOCl formulations in the pH range near the physiological pH of 7 are more suitable for medical applications, where bleach can be damaging and dangerous to users and the surfaces to which it is applied. Therefore, maintaining a pH between 4 and 9 is crucial to avoid damage to biological tissues or fragile surfaces.

[0045] In therapeutic technology, an active pharmaceutical ingredient (also referred to herein as "API") is understood to be the chemical component(s) that produce a respective therapeutic effect. A weakness of prior art formulations is that when hypochlorous acid is the only active API, there is no biocompatible activator and pH stabilizer that simultaneously acts as both a buffer and an antimicrobial agent, and preferably also exhibits a synergistic effect with the API.

[0046] Another weakness of prior art technologies is that most of the aqueous solutions in these cases are produced by electrolysis of isotonic 0.9% NaCl / H2O (normal saline), which lacks buffering capacity due to the absence of a mixture of biocompatible weak organic acids and their salts. Furthermore, electrolysis is a rather complicated and sometimes inconvenient method for preparing antibacterial solutions (i.e., in battlefields, tourist destinations, or areas of major disasters or pandemics).

[0047] A further weakness of prior art chlorine oxide-based formulations is their lack of stability, due at least in part to the fact that chlorine oxide gradually deteriorates and decomposes within the first few months of storage unless stored at low temperatures and in the absence of light and oxygen. Thus, pharmaceutical shelf-life stability of chlorine oxide solutions cannot be achieved at ambient conditions.

[0048] Yet another weakness of prior art chlorine oxide-based formulations is the lack of a controlled ionic strength compatible with the use of a given formulation. Ideally, administration of any given formulation containing chlorine oxide to a mammal requires an ionic strength of 300 mOsm to be isotonic with body fluids, which is equivalent to a concentration of approximately 150 mM NaCl.

[0049] Hypochlorous acid may be an attractive compound to include in antibacterial solutions because it is naturally produced in pure form by neutrophils in vivo. Furthermore, circulating monocytes, tissue-resident macrophages, and microglia in mammals also produce hypochlorous acid to inactivate pathogens within phagocytic vacuoles and in the extracellular space surrounding phagocytes in tissues. Thus, hypochlorous acid is a natural biological compound produced in relatively high concentrations by mammals. Activated neutrophils can mobilize their primary granule enzymes to the cell surface. For example, as described by Hirche et al., J. Immunol. (2005); 174:1557-1565, the enzyme myeloperoxidase (MPO) in activated neutrophils uses hydrogen peroxide (HO) catalyzed by NADPH oxidase to generate HOCl.

[0050] Given the large numbers of neutrophils that accumulate in inflamed tissues, such as the lungs of patients with acute respiratory distress syndrome, the concentration of HOCl that may be clinically useful is particularly physiologically relevant. 6 It is well known in the art that activated neutrophils can produce as much as 100 μM of HOCl within 2 hours, and therefore mammalian biological systems can handle concentrations of at least 100 μM of HOCl without pathological consequences.

[0051] Another chlorine oxide useful as an API in antibacterial formulations is chlorine dioxide, in which the chlorine atom is in the oxidation state +3. The primary reaction of sodium chlorite is the production of chlorine dioxide, as shown in the following equation (Equation 2): [ka]

[0052] Referring to the above equation (Equation 2), the HOR is typically a mineral acid such as HCl or citric acid, since a proton source is required to first convert sodium chlorite to chlorous acid and then to chlorine dioxide, a gas that is highly water soluble at room temperature.

[0053] One advantage of chlorine dioxide is that it cannot generate chlorine gas, Cl2, which is known to react with chlorinated hydrocarbons, such as trihalomethanes, which are toxic environmental pollutants. Another advantage of chlorine dioxide is that its disinfectant activity and the stability of its aqueous solutions are not pH dependent.

[0054] Chlorine dioxide, produced from sodium chlorite, is approved by the FDA for water disinfection under some conditions and is used to wash fruits, vegetables, and poultry. Sodium chlorite, NaClO2, is a solid precursor to chlorine dioxide and is sometimes used in combination with zinc chloride. It is also used as an ingredient in therapeutic rinses, mouthwashes, toothpastes and gels, mouth sprays, as a preservative in eye drops, and in contact lens cleaning solutions under the brand name Purite.

[0055] Chlorine dioxide is also used for bleaching and stripping textiles, pulp, and paper. It is also used to disinfect municipal water treatment plants after conversion to chlorine dioxide. Chlorine dioxide is used to sanitation food-contact hard surfaces and as a cleaner or rinse for various foods, including red meat, poultry, seafood, fruits, and vegetables. Because chlorine oxide compounds are unstable even when properly prepared, there are no measurable residues in foods after disinfection. Chlorine dioxide is also used as a teat dip to control mastitis in dairy cows.

[0056] The U.S. Army Natick Soldier Research, Development and Engineering Center has developed a portable, "powerless" method for producing chlorine dioxide, known as ClO2 gas, an effective biocide that can be used to combat contaminants ranging from harmless microorganisms and food pathogens to Category A biological terrorism weapons. Weeks after the 9 / 11 attacks, when anthrax was delivered in letters to civil servants, hazardous materials disposal teams used ClO2 to decontaminate the Hart Senate Office Building and the Brentwood Mail Facility.

[0057] In response to the COVID-19 pandemic, the U.S. Environmental Protection Agency included ClO2 as an agent that meets its criteria for use in environmental response to the coronavirus that caused it (see US EPA, OCSPP (March 13, 2020), "List N: Disinfectants for Use against SARS-CoV-2" (retrieved March 28, 2020, from the US EPA); "How we know disinfectants should kill the COVID-19 coronavirus" (Chemical & Engineering News, retrieved March 31, 2020).

[0058] However, these technologies also lack biocompatible activators and stabilizers of the chlorine oxide precursors that simultaneously act synergistically in the antimicrobial activity of the final formulation, so that such formulations in these cases are also subject to the stability problems mentioned above.

[0059] In summary, the main problem with the prior art medical use of solutions of chlorine in oxidation states higher than -1 is that these species are in higher energy states and are often referred to as chloride ions, Cl. - The stability, or lack thereof, of chlorine oxide is due to its tendency to revert to its original state and decompose in solution at ambient temperatures. This hampers the shelf-life stability required for chlorine oxide pharmaceutical formulations and medical devices at ambient conditions. Therefore, it is difficult to achieve adequate shelf life for chlorine oxide solutions, as required for medical devices and drugs. This limitation, inherent in all oxides of chlorine, hampers transportation and storage at higher temperatures, especially in areas where temperature, light humidity, and atmospheric gases vary.

[0060] Effervescent powders, tablets, or granules typically contain solid acid and base components that, when mixed with water or a pharmaceutically acceptable carrier, produce carbon dioxide. Typical, non-limiting examples of acids useful in this reaction include mono-, di-, or tri-acids, such as propionic acid, lactic acid, succinic acid, glutaric acid, pyruvic acid, citric acid, malic acid, oxaloacetic acid, tartaric acid, adipic acid, fumaric acid, or heptanedioic acid or octanedioic acid or higher diacids, or their derivatives. Citric acid imparts a citrus-like flavor to the product. However, acids that cannot be oxidized by chlorine species (e.g., but not limited to, succinic acid, glutaric acid, pyruvic acid, oxaloacetic acid, and adipic acid) are preferred acids in the present invention.

[0061] Typical bases used in effervescent reactions are sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate. Sodium bicarbonate is very common in effervescent formulations, producing a clear solution after tablet disintegration. Both types of carbonates are primarily used as desiccants, contributing to a drier product.

[0062] A water-soluble binder is usually required in effervescent tablets to make the tablet hard enough to be handled, and non-limiting examples of binders are monosaccharides, disaccharides, polysaccharides, or monohydroxy acids or polyhydroxy acids. Particularly preferred binders are dextrose, lactose, sorbitol, ascorbic acid sorbitol, or mannitol. The above-mentioned carboxylic acids can also be used as lubricants. The ideal amount of binder is the amount that makes the tablet hard enough to be handled, soft enough to disintegrate, and dry enough to be stable. The harder the tablet, the slower it disintegrates.

[0063] Lubrication of effervescent tablets can contribute to the efficient and efficient production of tablets or granules. Non-limiting examples of water-soluble lubricants are dextrose, lactose, sorbitol, ascorbic acid, sorbitol, mannitol, sodium benzoate, potassium sorbate, polyethylene glycol or its derivatives, succinic acid, adipic acid, glutaric acid, or clavulanic acid. Tablet presses that use a lubricating spray on the punches may not require lubrication.

[0064] Depending on the product, formulators may use colors (artificial or natural), sweeteners (acesulfame potassium, saccharin sodium, aspartame, and sucralose), and flavors (artificial or natural) to enhance the product or to mask the taste or odor from the active ingredient.

[0065] Effervescent tablets and powders are manufactured in much the same manner as conventional tablets and powders, but production should preferably be carried out in a temperature-controlled, very low humidity area. Effervescent granulations can be mixed in conventional mixing equipment such as ribbon, twin-cone, and V-type blenders. All equipment should be well grounded, and the product should be dried after cleaning.

[0066] Wet granulation of effervescent bases or fluid bed dryers can be attractive manufacturing methods, in which the effervescent mixture is suspended in a current of hot, dry air while being sprayed with a solution of water or binder.

[0067] Vacuum granulators have also been used to produce effervescent granules. In this method, water or a binder solution is sprayed onto the effervescent mixture while blending. The granules are placed under vacuum and dried by heating via a heating jacket.

[0068] The advantages of using effervescent tablets or granules are many. One is self-mixing with CO2, which often eliminates the need for tedious shaking procedures. Another advantage is the flexibility in including flavors, colors, and thickeners in the material. Yet another advantage is the pre-calculation of the amount of precursor and prodrug of the active ingredient in the effervescent material.

[0069] The present invention recognizes all these deficiencies associated with prior art compositions that use chlorine oxides. In particular, the present invention provides compositions that include a solid precursor of oxidized chlorine (OC) in combination with an activator that provides a source of protons.

[0070] In addition to chlorine oxide, as defined above, two activators are included in the foamable material: The first activator is a solid carboxylic acid or salt thereof having the formula RN(COOH)m, where RN is a saturated and unsaturated organic moiety containing 1 to 25 carbon atoms, optionally substituted with oxygen and hydrogen to form a functional group that cannot be oxidized by chlorinated species, and m is an integer from 0 to 10 (hereinafter, acid activator).

[0071] The second activator is a base that can react with the acid activator to produce carbon dioxide. The second activator has the following general formula: (M n+ ) s ((H) t CO3) u (wherein n, s, t, and u are integers ranging from 1 to 2, s from 1 to 2, t from 0 to 1, and u from 1 to 2, such that the sum t+s+n=1 to 5; and M is an alkali metal, alkaline earth metal, or transition metal).

[0072] Preferred basic activators are sodium or calcium carbonate or sodium bicarbonate.

[0073] Such compositions are disinfectants useful for treating a wide range of microorganisms. In particular, when the active pharmaceutical ingredient is generated at the site of use from a stable solid precursor of chlorine oxide, hereinafter referred to as "API-P," and an acetic acid activator is included to buffer the solution or gel to a biocompatible pH value while reacting, the stability problems of the prior art no longer exist.

[0074] As previously mentioned, the prior art solutions do not address how to ensure the ionic strength or osmolality of the final antimicrobial solution is biocompatible with biological fluids. Furthermore, the prior art does not demonstrate how to adjust and increase the contact time and duration of the API in the area of ​​therapeutic interest, for example, by adjusting rheology and flow properties. Furthermore, the prior art does not provide a relatively simple yet effective means of monitoring the oxidation state of the API, nor a visual indication of where the API has been applied during mixing of the disinfectant composition.

[0075] Additionally, in some embodiments, the compositions of the present invention may further include the use of a thickening agent (also referred to herein as a "VE") and / or may include a combination of a solid precursor of oxidized chlorine with an activator, such as a diacid or salt thereof.

[0076] Another aspect of the present invention is the inclusion in the formulation of a dye, preferably a redox-sensitive dye, whose color changes with the oxidation state of the chlorine atom, an advantage that addresses the drawbacks described with prior art compositions.

[0077] In particular, preferred compositions of the present invention are in solid form, thereby eliminating any shelf-life related problems encountered with hypochlorous acid or chlorine dioxide solutions described in the prior art.

[0078] More specifically, the immediate generation of a ready-to-use formulation of chlorine oxide species from the solid precursor API-P can be released from effervescent powders, tablets, or granules. The effervescent powders, tablets, or granules can be used for the preparation, distribution, and long-term stable storage of the compositions prepared in accordance with the present invention. In particular, such effervescent tablets or granules described herein can have some of the components required to produce the compositions of the present invention.

[0079] In an example, a solid precursor of oxidized chlorine and an activator, such as succinic acid, glutaric acid, adipic acid, pyruvic acid, or salts thereof, a thickener, and a dye are mixed, after which the composition forms in the desired formulation of a disinfectant at the desired time and place of use.

[0080] Furthermore, in addition to their antimicrobial properties, the acids exemplified above are attractive because they cannot be further oxidized by oxidizing agents such as OC and because of their endogenous nature, which is biochemically speaking high in concentration.

[0081] Thus, effervescent powders, tablets, or granules allow practical use in immediately mixing the necessary ingredients to produce an active solution of the API at the point of use. Note that in medical applications, a pre-calculated amount of NaCl can additionally be used in the preparation of the effervescent tablets, depending on the planned use, to ensure the ionic strength or osmolality of the final antimicrobial solution matches the osmolality in the area of ​​use.

[0082] A preferred embodiment of the present invention is the inhalation of a selected solution of the present invention to combat viral infections in the respiratory system of a mammal. Therefore, any nebulizer or inhaler that converts a liquid into an aerosol and is commonly used for the treatment of cystic fibrosis, asthma, chronic obstructive pulmonary disease (COPD), and other respiratory diseases or disorders is useful in the present invention. The devices often use compressed air or ultrasonic energy to atomize the disinfectant solution. Any type of pressurized metered-dose inhaler (pMDI), dry powder inhaler (DPI), or slow mist inhaler (SMI) is particularly useful, as described, for example, by Prajapati et al. in IJPSR, 2019; Vol. 10(8):3575-3582. Any electrostatic or non-electrostatic inhaler, such as VORTEX, Pari, or Sympotec, is also useful in practicing the present invention.

[0083] The effervescent powder, tablet, or granules have no stability issues and produce a broad-spectrum antimicrobial solution upon mixing of the ingredients, leaving only biocompatible, inert chemical species already found in human biology or nature.

[0084] As previously mentioned, activation of the API is accomplished using an activator (e.g., succinic acid or pyruvic acid) that acts synergistically with chlorine oxide against microorganisms and further maintains acidity in a controlled pH range between pH 4 and pH 8. This novel method and its formulation avoids the inherent lack of long-term stability of chlorine oxide OC in solution (due to its oxidizing potential) because it does not require storage of the disinfecting composition as an aqueous solution prior to use.

[0085] Another advantage of the present invention is the option to add other compounds to aid application. For example, in wound healing applications, it is necessary to increase the viscosity (μ) of the product on the skin to extend contact time. The present invention solves this problem through the use of a water-soluble or soluble thickener (VE) that cannot be chemically oxidized by the API, thereby providing improved control of the contact time and duration of the API in the area of ​​therapeutic interest. The VE ensures that the rheology and flow properties are tailored to each method and disinfection area to create a solution or gel with perfect flowability. The VE can include a water-soluble gelling agent, such as polyacrylic acid, polyethylene glycol, or any other oligomer or polymer that cannot be oxidized by the API.

[0086] Additionally, the composition may contain one or more dyes identified from the group of redox dyes (also referred to herein as "ROD" or "RODs"), whose color and intensity depend on the oxidation state of chlorine oxide. It is noted that in addition to providing a visual indication (i.e., by color) of the oxidation state of the chlorine atom, RODs also provide their own antimicrobial efficacy. This enhances the synergy between the ingredients in the formulation in a novel manner. The ROD can maintain its color for a period of time sufficient to monitor the oxidative activity of the API, chlorine oxide, and also provide a visual indication of the area to which the formulation has been applied, thereby addressing shortcomings of the prior art.

[0087] Advantages and further inventive features of the present invention will become apparent from the description of the invention provided below.

[0088] Chlorine oxide species (denoted as OC) have the general formula shown below: M n+ [Cl(O) x ] n n- wherein M can be any alkali metal, alkaline earth metal, or transition metal ion; n can be an integer from 1 to 5; x can be an integer from 1 to 4; and y can be an integer from 1 to 2.

[0089] When M=Na, n=1, x=1, API-P is NaOCl5. When M=Ca, n=2, x=1, API-P is solid Ca(OCl)2. When M=Na, n=1, x=2, API-P is solid NaClO2. When M=Ca, n=2, x=2, API-P is solid Ca(ClO2)2. When x=3 or 4, API-P produces more reactive chlorate and perchlorate species.

[0090] One non-limiting example is the instant generation of hypochlorous acid from sodium or calcium hypochlorite mixed with monosodium succinate in cap 2 according to FIG. 1 to provide a ready-to-use solution of API hypochlorous acid at pH 4-6, optionally containing colorants and thickeners.

[0091] Another non-limiting example is calcium hypochlorite, Ca(OCl)2, a stable, water-soluble API-P of HOCl, which is produced and sold in ton-scale as a pool disinfectant. Calcium hypochlorite dissolves rather slowly in water, leaving only calcium hydroxide, which is found in nature and living organisms and is used in food products under the E number E526, to produce HOCl, one of the two active ingredients in this invention. HOCl is Cl. - and biocompatible species containing hydrogen and oxygen. However, according to the present invention, the effervescent generation of CO2 greatly increases the dissolution rate of Ca(OCl)2 and other solid components.

[0092] Another preferred embodiment of the present invention is the solid precursor of chlorine oxide, tetrachlorodecaoxide (TCDO), CAS number 92047-76-2, also known as a stabilized solution of WF10 or OXO-K993, prepared as described by Meuer et al., Canadian Patent No. 2616008. Tetrachlorodecaoxide (TCDO) is a chlorite ion, ClO2. - can be prepared by combining the alkali or alkaline earth salt of with excess oxygen in water.

[0093] Therefore, one advantage of the present invention is that there is no pharmaceutical stability problem in the solid form of the precursor API-P in a dry, water-free quality, and therefore the present invention solves one of the major technical problems in the prior art.

[0094] A key aspect of the present invention is the combination of API-P with a molecule containing a carboxylic acid functionality -COOH, a sulfonic acid functionality -SO3H, a phosphoric acid functionality -PO3H, or a boric acid functionality -B(OH)2, defined below as an activator of API-P in the formulation. The activator has the general formula R1XO n (R 2, ) m where the group R1 can be a group containing 1 to 10 hydrogenated carbon atoms, optionally substituted with amino, amido, carboxylic acid, or hydroxy groups. The group X can be a carbon, phosphorus, or sulfur atom, n and m can be integers 2 or 3, and R2 can be a proton, H, or any alkali metal, alkaline earth metal, or transition metal ion. The nature of the substituents in the formula varies depending on the application and chlorine species, and can be any compound containing an amino group, such as ammonia, an amino acid, such as taurine, or a therapeutic agent that enhances the synergistic potential of the formulation. Activators have the general formula R1XO n It may be any combination or mixture of two or more compounds defined by R2.

[0095] A preferred, non-limiting example is the carboxylic acid R3(COOH)2, where R3 is a straight or branched saturated or unsaturated hydrocarbon chain having 1 to 24 carbon atoms. Non-limiting examples of activators can be succinic acid, glutaric acid, adipic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, hippuric acid, maleic acid, boric acid, sulfuric acid, phosphoric acid, or boric acid.

[0096] Taurine is particularly preferred because it is an endogenous amino acid that normally moderates the effects of OCs in the body and can combine with OCs to form endogenous N-chloro-amino acids such as ClNH-CHCH-SOH, which themselves have antibacterial properties.

[0097] Succinic acid, glutaric acid, and pyruvate are even more preferred because they are endogenous substances, have antibacterial properties, have very low toxicity, and form buffer solutions in mixtures with their metal salts, and are used as non-limiting examples in the further description of the present invention. For example, Anuj Purohit and Anand Mohan reported in LWT-Food Science and Technology (2019), 116108596, pp. 1-7 that pyruvate and monosodium succinate (mixture), individually or in mixture, reduced the log CFU of bacteria per gram of ground chicken by approximately log 2.

[0098] An advantage of the present invention is that since API-P is a solid and commercially available on a large scale, the solid multi-component product according to the present invention is not hampered by stability issues in the environment of a pharmaceutical product or medical device, regardless of temperature, air, humidity, light, oxygen or other ambient conditions.

[0099] A further aspect of the present invention is that API-P does not dissolve instantly in water using shaking or stirring, but driven by the effervescent generation of CO2, API-P is instantly dissolved or suspended in the liquid phase, and a physiologically acceptable pH and ionic strength is quickly reached in the final solution in combination with succinic acid and / or its salts. All post-disinfection products are endogenous species existing in human biology or nature.

[0100] Another aspect is that the instant generation of active disinfectant at the point of use of the present invention has a significant and profound impact on the versatility of product applications, paving the way for much more flexible product packaging technology, as the size and amount of effervescent material in the initial package can be freely varied depending on the use.

[0101] Single-use two- or three-component effervescent packages containing small, stable, single doses are available, ideally suited for tourists, disaster zones, military personnel, or microbial pandemics. Additionally, API-P-containing bulk bag, box, or tank designs suitable for disinfecting larger areas are useful in agricultural settings, the aquaculture industry, or military operations.

[0102] Thickener for the preparation of viscous solutions and gels A preferred aspect of the present invention is the option to include other compounds in the effervescent powder, tablet, or granule that aid in the ease of use of the product. For wound healing or skin disinfection applications, there is a need to increase the viscosity (μ) of the product on the skin to extend contact time. Thickener VE solves this medical need.

[0103] The preferred VEs of the present invention are water-soluble gelling agents in which all oxygen-containing functional groups in the VE are in their highest oxidation state, thereby preventing oxidation of the API, and providing long-term persistence of the API in the area of ​​interest (e.g., mammalian skin).

[0104] Examples of gelling agents according to the present invention include, but are not limited to, polyacrylic acid (CARBOMER), polyethylene glycol or any other oligomer, polymer or block copolymer thereof. Further, the thickening agent may be selected from poly(acrylic acid)-acrylamidoalkylpropane sulfonic acid copolymer, phosphinopolycarboxylic acid and poly(acrylic acid)-acrylamidoalkylpropane or sulfonic acid sulfonated styrene terpolymer.

[0105] A preferred VE is polyacrylic acid because each monomer contains a carboxylic acid functional group. Therefore, since the pKa of PAA is 4.5, using this VE adds to our preferred pH range of between pH 4 and 5.5.

[0106] Therefore, a preferred aspect of the present invention is the use of polymers such as acrylate copolymers of any type known to those skilled in the art, which can function well in the formulations of the present invention at concentrations ranging from 0.01 to 5%. Acrylate copolymers are homopolymers and copolymers of acrylic acid crosslinked with polyalkenyl polyethers. Acrylate copolymers vary in graft density; they differ in oxidative capacity and the number of grafted chains per polymer. One possible crosslinker is pentaerythritol, which is highly stable and therefore suitable for use in the present invention. Polyacrylic acid (PAA) polymers, known to stabilize H2O2 formulations, can be used in the present invention (see Schmucker-Castner & Desai, 1999, "Rheology Modification of Hydrogen Peroxide-Based Applications Using a Cross-linked PAA Polymer," Int J Cosmet Sci 21(5):313-25).

[0107] The polymer-stabilized solutions of OCs according to the present invention have applications in many situations, such as wound treatment, aseptic packaging, electronics manufacturing, and pulp and paper bleaching. Formulations of APIs are compatible with formulation as gels or viscous fluids and can be applied to either inanimate target surfaces or target surfaces representative of infected epithelial mucosa or skin surfaces of infected humans or animals to ensure prolonged, intimate contact with the required level of API. Non-viscous formulations of APIs can also be dispersed into the air in confined spaces as mists to achieve environmental disinfection or for inhalation purposes to treat respiratory diseases.

[0108] In an example, polyacrylic acid carbomer has increased viscosity at concentrations of 0.01-0.1%. If desired, polyacrylic acid carbomer forms a regular gel at concentrations ranging from 0.1-1%. Depending on the solution, a mixer, stirrer, or sonicator must be used to dissolve VE in the microbicide formulation. The degree of gelation can be further increased by using a biocompatible base (e.g., pharmaceutical buffer triethanolamine or other biocompatible amino alcohols).

[0109] Antibacterial redox-sensitive antibacterial dyes as indicators. A further innovative aspect of the present invention, and novel in the context of disinfection technology, is the use of redox dyes (hereinafter referred to as RODs for clarity), whose color and intensity depend on the oxidation state of the OC. Even more advantageously, the identified RODs have antibacterial effects, enhancing the antibacterial synergy between the components of the present invention. If the standard half-cell potential of the ROD is less positive than that of the OC, the color of the formulation is maintained as long as the OC is active. This allows color to be visualized in areas where the formulation is applied and in areas where active OC is present. This is particularly advantageous when formulations according to the present invention are used in the treatment of mastitis, for example, where large groups of cows need to be treated for mastitis, and the colored formulations according to the present invention allow visualization of which animals have been treated. Furthermore, the use of an opposite type of indicator, which appears color when the oxidative power of the OC is lost, is also useful.

[0110] Non-limiting examples of suitable dyes useful in the present invention are pH-independent dyes that are visible in the presence of OC. Preferred examples are N-phenylanthranilic acid (purple-red), N-ethoxychrysoidine (cyan), o-dianisidine (red), sodium diphenylamine sulfonate (red-purple), diphenylbenzidine (purple), diphenylamine (purple), and viologens that are colorless in the presence of OC but deep blue in the absence of OC.

[0111] Another preferred example is the pink pigment trisodium (4E)-3-oxo-4-[(4-sulfonato-1-naphthyl)hydrazono]naphthalene-2,7-disulfonate, which is more commonly called amaranth (E number 123).

[0112] Examples of pH-dependent dyes that are deep blue in the presence of an active OC but colorless in the absence of OC are sodium 2,6-dibromophenol-indophenol or sodium 2,6-dichlorophenol-indophenol, sodium o-cresol indophenol, thionin (synonym: Rous violet), methylene blue, Gentian violet, indigo tetrasulfonic acid, indigo carmine (synonym: indigo disulfonic acid), and indigo monosulfonic acid. Examples of dyes that are red or red-purple in the presence of OC are phenosafranine, safranine T, neutral red, and dialkyl-p-phenylenediamine (SPD, red-purple).

[0113] Many of these dyes have inherent antimicrobial effects, i.e., methylene blue (MB) and Gentian Violet (GV) and their combinations have been used as antimicrobial dyes in foams for wound dressings in combination with polymers such as polyvinyl alcohol or polyurethane, as described, for example, by Edwards in Advances in Wound Care (2016), 5, pp 11-19.

[0114] A particularly useful class of pigments useful in the present invention are microbial phenazines, which are colored, redox-active, nitrogen-containing aromatic compounds with metabolic, ecological, and evolutionary significance. See, for example, Chincholkar, S. & Thomashow, L. Microbial Phenazines: Biosynthesis, Agriculture and Health (eds. Chincholkar, S. & Thomashow, L.) 1-243 (Springer, 2014). All of these unique features make them attractive microbial metabolites useful as pigments in pharmaceutical formulations. To date, reports of over 100 natural phenazines and over 6,000 synthetic phenazines are available, which exhibit promising biological activities, including antibacterial, anticancer, antiparasitic, and insecticidal, as well as biocontrol properties.

[0115] An even more attractive class of phenazines includes bis-N-oxide phenazines, which have even stronger antibacterial properties than the parent phenazines. Most of these compounds are naturally occurring compounds produced by bacteria and are heteroaromatic N-oxide compounds, hereafter referred to as HANOX. HANOX compounds are useful in the present invention because they are redox dyes (RODS), and their color depends on the oxidation state of the OC. At the same time, prior art HANOX compounds have been shown to have broad-spectrum antibacterial activity, as described, for example, in U.S. Patent No. 3,822,265 to Leimgruber et al.

[0116] Furthermore, the phenazine derivatives provided in U.S. Patent No. 3,822,265 have a broad spectrum of fungicidal activity. In particular, certain phenazine derivatives have shown high activity against a wide variety of bacteria, yeasts, and fungi, such as Streptococcus agalactiae, Staphylococcus aureus, Escherichia coli, Corynebacterium pyogenes, Moraxella bovis, Pseudomonas aeruginosa, Candida albicans, and Microsporum canis. Therefore, phenazine derivatives are particularly useful in treating animal diseases of microbial origin in agriculture.

[0117] A surprising finding regarding these derivatives is the lack of adverse effects on body tissue under the conditions of use, making them particularly suitable for topical application, preferably in amounts ranging from 0.05% to 1.0% by weight of the composition.

[0118] These derivatives are particularly valuable for topical application, for example in solid or gel formulations including finely divided powders and granular materials, as well as in liquid formulations including solutions, suspensions, concentrates, tinctures, slurries and aerosols, creams, gels, jellies, ointments and pastes.

[0119] An even more preferred HANOX group useful in the present invention is described in WO2015063516A2 and WO2018109504A1 by Viktorsson et al. This technology describes RODs that possess redox properties and broad-spectrum antibacterial properties. As described by Viktorsson et al. in Bioorg.Med.Chem.(2017)25,pp2285-2293, this particular class of HANOX is particularly attractive because it exhibits lower toxicity in mammals compared to the compounds described in U.S. Pat. No. 3,822,265 and can be used with a lower risk of toxic reactions in infected hosts.

[0120] Methylene blue is another particularly preferred dye useful in the present invention because it is FDA approved as an excipient in drug formulations, it has antibacterial properties, and its effectiveness as a therapeutic agent can be enhanced using photodynamic therapy.

[0121] Effervescent tablets useful in the present invention FIG. 1 is a schematic diagram illustrating a schematic, non-limiting, exemplary effervescent tablet for extemporaneously generating a disinfectant solution of an API with excipients in an aqueous solution in accordance with embodiments and methods of the present invention.

[0122] Effervescent tablets are used in a variety of medical, household, and agricultural applications. In many of these applications, the antibacterial solution produced from the effervescent material is useful in the present invention and can be produced in a two- or multi-chamber bottle, bag, syringe, inhaler, hand disinfection device, spray bottle or flask, or tank constructed of hard or soft materials (e.g., plastic, rubber, waterproof paper, or metal). The therapeutic formulation can be easily activated from the device at the bedside, in the field, or in a home environment without complex mixing procedures, and can be stored at ambient temperature. It can be combined with an automated dispensing system, easily labeled, and automatically recorded via barcode.

[0123] The effervescent tablets of the present invention are composed of pre-calculated amounts of ingredients, can be designed to eliminate mixing errors and avoid unwanted exposure to patients and personnel, and meet Joint Commission and USO 797 ​​guidelines.

[0124] Use of the present invention for antibacterial purposes with photodynamic therapy Bacterial elimination using antimicrobial photodynamic therapy (aPDT) has been demonstrated as an alternative treatment modality in the treatment of peri-implantitis. In Photodiagnosis and Photodynamic Therapy (2019), 25, pp. 7–16, Huang et al. described the dose- and pH-dependent bactericidal effect of methylene blue (MB)-mediated aPDT in eliminating Gram-negative bacteria (P. gingivalis and A. actinomycetemcomitans) and Gram-positive bacteria (S. mutans) on titanium alloys pretreated with sandblasting, grit, and acid etching (SLA). However, the test formulation did not contain OC to further enhance the therapeutic effect.

[0125] Therefore, another preferred embodiment of the innovative formulations comprising OC, succinic acid or its salts, and optionally a thickener, is the inclusion of a ROD, exemplified by methylene blue, for use in photodynamic therapy, for example to improve wound healing or bacterial infections in mammals, in which case the site of administration of the product according to the invention can be irradiated with light having a wavelength adapted to produce the photodynamic effect of the dye.

[0126] In Photodiagnosis Photodyn. Ther. (2018), 23, pp. 347-352, Souza et al. used photodynamic therapy to demonstrate the antibacterial activity of hypochlorite solutions and rotary instruments associated with photodynamic therapy on root canals infected with Enterococcus faecalis. However, the test solutions lacked antibacterial dyes. These techniques are incorporated herein by reference.

[0127] Use of the invention in agriculture In agriculture, especially in livestock farms, many kinds of infectious diseases caused by bacteria, viruses and fungi affect the daily operations of the farm and affect the cost of running the facility. In these situations, the formulations designed by the present invention act therapeutically or preventively, and are particularly useful in skin infections.

[0128] One important example is bovine mastitis, which costs the U.S. dairy industry approximately $1.7-2 billion annually. Effective and environmentally friendly treatment of mastitis has proven difficult because milk from cows long-term fed antibiotics is not marketable until residual drugs leave their systems. Because infections within a cow's udder and teats are away from the animal's main bloodstream, vaccines are ineffective. To mark treated cows, dairy workers apply strips of tape to warn and mark treated cows.

[0129] Thus, a preferred aspect of the present invention is the treatment of mastitis using a gel or viscous solution containing OC, succinic acid or its salt, or one of the other preferred acid-activating agents listed above, a thickening agent VE, and a ROD, exemplified by methylene blue. The colored gel remains in the breast and nipple area, the succinic acid has the ability to penetrate into the nipple skin, and the color eliminates the need for tape. Furthermore, the applied gel can be irradiated with light of an appropriate wavelength to enhance the gel's therapeutic effect. In this case, steps 1 through 4 and step 6 are performed to provide a useful, immediate formulation.

[0130] Use of the invention in aquaculture Water quality is a prerequisite for successful aquatic animal farming, exemplified by fish, oysters, shrimp, and prawns. Open water systems often introduce organisms such as viruses, bacteria, lice, protozoa, fungal pathogens, algae, and parasites. Common viral infections that cause high mortality in aquatic species attractive for food production are carp herpesvirus disease, pancreatic disease (PD), and infectious salmon anemia (ISA). Adequate water quality or sufficient quantities of clean water are often unavailable. Prior art breeding facilities often lack the means to prevent these infectious species from accessing and affecting the breeding species. Furthermore, once infection has occurred, no efficient treatments exist to provide effective cure for these diseases.

[0131] A preferred embodiment of the chlorine dioxide species OC of the present invention is the effective treatment of all these infectious diseases and pests and cells. Extemporaneous formulations of OC are highly effective in suppressing these waterborne pathogens. In an example, chlorine dioxide is a broad-spectrum biocide effective in solving a distinct problem in the prior art. The formulations of the present invention are destructively effective against disease-causing microorganisms without harming the gills or any other parts of the fish or captive species, even being used in special tanks for repeated treatment of captive salmon, for example. For these applications, a series of preparations in which the API-P is NaOClO2 or Ca(OClO2)2 are mixed with a pre-calculated amount of succinic acid in steps 1-3 and used in effervescent tablets.

[0132] Anti-weapon applications of the present invention Another preferred embodiment of the oxidizing chlorine species OC of the present invention is the reduction or elimination of nerve gases such as G series exemplified by GA, GB (sarin), GD (soman), GF (cyclosarin), and GV, V series exemplified by VE, VG, VM, VR, and VX, Novichok, carbamate, insecticides, protein toxins such as ricin, or warfare agents such as bacteria, viruses, or fungi for biological warfare.

[0133] All of these species share the common feature of being oxidizable by the oxidizing chlorine species of the present invention. For example, the nerve agents GD or soman have a phosphorus halide group that can be oxidized to harmless species by oxidizing chlorine species, as described by Xu et al. in E3S Web of Conferences 267, 02043 (2021), pp. 1-6.

[0134] However, in this example, an unstable hypochlorous acid solution was used. The stability problem was solved in accordance with the present invention by using a solid foaming material as the instant precursor to the reactive solution.

[0135] One example of an anti-warfare use of the solutions produced in this invention is the generation of a mist or fog of oxidizing aerosol droplets that can persist indoors or outdoors for extended periods of time to protect regional or local organisms from warfare agents.

[0136] Antiviral Uses of the Invention Additionally, the methods disclosed herein allow for improved methods of exposing contaminated surfaces, equipment (e.g., medical devices), any furniture surface, door handle, device, clothing, or personnel to disinfectant formulations using spraying or vaporization of APIs into confined spaces. This is possible because the breakdown state and quality of the disinfectant formulation is known because it is not stored as a solution but is prepared fresh from solid precursors at the site of interest.

[0137] This procedure ensures dispersion of the active agent into crevices and microenvironments, even on personnel suspected of being contaminated with infectious tissue or bodily fluids. Vaporization of these formulations may enable beneficial therapeutic or prophylactic effects against resistant viral, bacterial, or fungal infections.

[0138] Kim et al., in Laryngoscope (2008), 118, pp. 1862-1867, investigated the effects of low-concentration hypochlorous acid for nasal irrigation on bacteria, fungi, and viruses. The formulation was also used in vitro against human rhinovirus (HRV) in nasal epithelial cells, demonstrating significant virus-killing effects. As described by Hakim et al. in J. Vet. Med. Sci. (2015), 77, pp. 211-215, a similar formulation was used in vitro against avian influenza virus, demonstrating favorable effects.

[0139] Hypochlorous acid has also been used clinically on the upper respiratory tract mucosa of patients with rhinitis. Cho et al. reported improved outcomes after low-concentration hypochlorous acid nasal irrigation in pediatric chronic sinusitis in Laryngoscope (2016), 126:791-795. To the best of our knowledge, similar formulations have not been used in the prior art to treat other parts of the respiratory system in mammals.

[0140] Robert Northey, in U.S. Patent No. 10,342,825, describes a low pH antibacterial solution containing 5 mg / L to 200 mg / L hypochlorous acid and water, the solution having a pH of 5.6 and stabilized by a phosphate buffer. The solution can be vaporized using a nebulizer for distribution on surfaces and tissues. However, the drawbacks of the formulations used in the prior art are the same as those described above.

[0141] WO 2019 / 222768 by Terry describes a method for inactivating infectious agents that are resistant viruses, oncogenic viruses, chemically resistant non-enveloped viruses, or infectious agents present on mucosal or epithelial surfaces. The formulation contains the infectious agent along with an unbuffered electrolyzed hypohalous acid composition. However, in this case, the weaknesses of the formulation used in the specification are the same as those defined above.

[0142] Many of the problems in the prior art have now been solved with the present invention: for the first time, OC has been combined with an activator from a solid precursor and NaCl to maintain biological osmolality for antiviral use of the resulting formulation.

[0143] Exposure can be achieved without the toxicity and corrosiveness concerns associated with prior art methods of inactivating highly transmissible and resistant infectious agent types. A preferred aspect is to eradicate, minimize, or prevent the progression of a viral infection in the upper respiratory tract, so that the immune system has time to mount an antibody response against the virus.

[0144] Thus, the systems and methods of the present invention provide chlorine oxide OCs as a means of treating viral infections in the respiratory tract. The compositions of the present invention are capable of treating other infectious diseases, including but not limited to SARS, MERS, and SARS CoV-2 infections. This is facilitated for the first time through the immediate precursor of the API combined with the effervescent tablet of the present invention, as there is no need to evaluate the lack of activity of a solution stored at ambient conditions.

[0145] In particular, inhalable hypochlorous acid formulations of OC; activators, such as succinic acid or pyruvic acid; excipients for adjusting the rheology of the final solution; osmolality adjusters, such as sodium chloride - such ready-to-use formulations can now be prepared on-site, along with delivery methods via nebulizers, such as soft mist inhalers, jet nebulizers, ultrasonic nebulizers, and vibrating mesh nebulizers. When used, inhalers and nebulizers aerosolize the compositions of the present invention for delivery by inhalation.

[0146] The formulations used to generate aerosols can be provided in dry powder, solution, or suspension form. Fine droplets, sprays, and aerosols can be delivered by intranasal or intrapulmonary pump dispensers or squeeze bottles. The compositions can also be inhaled through inhalers, such as metered-dose inhalers or dry powder inhalers. The compositions can also be inhaled through nebulizers, such as ultrasonic nebulizers, which deliver the OC and succinic acid composition directly to the airways via an inhalable formulation. This prevents and treats respiratory infections caused by viruses and other microorganisms. According to the present invention, the formulations described herein are safe and effective for preventing and treating viral infections.

[0147] The composition of the present invention can also contain a pharmaceutically acceptable carrier, such as a diluent, to facilitate delivery to the respiratory mucosa.The carrier can be an aqueous carrier, such as water or saline.The composition can be isotonic, having the same osmotic pressure as blood and tears.Suitable non-toxic pharmaceutically acceptable carriers are known to those skilled in the art.Various carriers can be particularly suitable for different formulations of the composition, for example, whether it is used as a drop, a spray, a suspension, or another form for pulmonary delivery.

[0148] The preparation for inhalation can be provided in dry powder form, solution or suspension form.The composition can be delivered by various devices known in the art for administering drops, droplets and spray.The composition can be delivered by dropper, pipette or dispenser.Fine droplets, spray and aerosol can be delivered by intranasal or intrapulmonary pump dispenser or squeeze bottle.

[0149] Intranasal delivery can be provided via a nasal spray device. Thus, the formulation according to the present invention can be designed as a nasal spray, which is blown into the nose and delivered to the respiratory tract.

[0150] The soft mist inhaler uses mechanical energy stored in a spring, actuated by the user, to apply pressure to a liquid container, causing the contained liquid to spray through a nozzle for inhalation in the form of a soft mist. The soft mist inhaler does not rely on a gas propellant or electrical power for operation. The average droplet size in the soft mist inhaler is approximately 5.8 micrometers.

[0151] Jet nebulizers are the most commonly used and are sometimes called atomizers. Jet nebulizers use compressed gas (e.g., air or oxygen) to aerosolize liquid medications when released at high velocity from the jet nebulizer. The resulting aerosolized droplets of therapeutic solution or suspension are then inhaled by the user for treatment. The compressed gas can be pre-compressed in a reservoir or compressed on demand by a compressor within the nebulizer.

[0152] Ultrasonic nebulizers rely on an electronic oscillator to generate high frequency ultrasound waves that, when directed through a reservoir of a liquid therapeutic suspension, aerosolize the medication for inhalation.

[0153] Vibrating mesh nebulizers use the vibration of a membrane with thousands of holes on top of a liquid reservoir to aerosolize a fine droplet mist for inhalation. Vibrating mesh nebulizers avoid some of the drawbacks of ultrasonic nebulizers, offering more efficient aerosol production with shorter treatment times and less heating of the liquid being nebulized.

[0154] The treatment of viral infection is achieved by using the synergistic composition of glutaric acid and hypochlorous acid.Glutaric acid component is particularly effective in penetrating into tissue, while hypochlorous acid is particularly effective in treating the infection on the outer surface of tissue.As mentioned above, these compositions are effective in treating airway and preventing respiratory infection.

[0155] The disclosed compositions are particularly effective because balancing the concentrations of hypochlorous acid and glutaric acid with NaCl allows for safe treatment of viruses. The exact balance depends on the formulation, the treatment site, and the desired amount of surface penetration. Hypochlorous acid can be present at about 5 ppm up to about 1000 ppm or more. Different applications, delivery methods, and tissue types may require higher or lower concentrations. Glutaric acid can be present at about 0.1% up to about 5.0% or more, preferably about 1.0%. By balancing the two components, the composition can have the dual effect of treating both the surface and subsurface of the tissue to which it is applied.

[0156] When the OC is hypochlorous acid (HOCl), a ready-to-use composition having an OC concentration of about 15-200 ppm is usually sufficient to treat infected lungs. When the OC is chlorine dioxide (OCl2), a concentration of 0, 1-5 ppm is usually sufficient.

[0157] In some cases, to completely destroy the virus or prevent it from entering the respiratory tract, the composition should be in contact with the virus for an extended period of time, ranging from a few seconds to a few minutes to an hour or more. Thus, in some embodiments, the composition is in the form of a gel, which allows for longer contact time with the infected site.

[0158] The use of the composition in combination with known antiviral treatments can increase the effectiveness of the composition.In some embodiments, the method of the present invention further comprises administering one or more doses of antiviral substances (simultaneously or sequentially with the composition of the present invention).These can include, but are not limited to, acyclovir, adefovir, adamantine, boceprevir, brivudine, cidofovir, emtricitabine, entecavir, famciclovir, fomivirsen, foscarnet, ganciclovir, lamivudine, penciclovir, telaprevir, telbivudine, tenofovir, valacyclovir, valganciclovir, vidarabine, m2 inhibitor, neuraminidase inhibitor, interferon, ribavirin, nucleoside reverse transcriptase inhibitor, non-nucleoside reverse transcriptase inhibitor, nonstructural protein 5a (ns5a) inhibitor, chemokine receptor antagonist, integrase strand transfer inhibitor, protease inhibitor and purine nucleoside.

[0159] The compositions are also useful in combination with known antibacterial treatments.

[0160] In some embodiments, the methods of the present invention further comprise administration (concurrently or sequentially with the compositions of the present invention) of one or more doses of an antibiotic including, but not limited to, a beta-lactam antibiotic such as ciprofloxacin, ampicillin, or a carbapenem, azithromycin, cephalosporin, doxycycline, fusidic acid, gentamicin, linezolid, levofloxacin, norfloxacin, ofloxacin, rifampin, tetracycline, tobramycin, vancomycin, amikacin, ceftazidime, cefepime, trimethoprim / sulfamethoxazole, piperacillin / tazobactam, aztreonam, meropenem, colistin, or chloramphenicol.

[0161] In some embodiments, the methods of the invention further comprise the administration of one or more doses of an antibiotic from an antibiotic class including, but not limited to, aminoglycosides, carbacephems, carbapenems, first-generation cephalosporins, second-generation cephalosporins, third-generation cephalosporins, fourth-generation cephalosporins, glycopeptides, macrolides, monobactams, penicillins, polypeptides, quinolones, sulfonamides, tetracyclines, lincosamides, and oxazolidinones. In some embodiments, the methods of the invention comprise the administration of a non-antibiotic antibacterial agent including, but not limited to, sertraline, racemic and stereoisomeric forms of thioridazine, benzoyl peroxide, taurolidine, and hexetidine.

[0162] The dosage regimen of the composition can include the amount, frequency, and duration of exposure to the composition. The dosage regimen can depend on the severity of the infection or the regimen prescribed for treating or preventing the viral infection.

[0163] The composition can be administered in one dose or multiple doses per day, for example, 2, 3, 4 or more doses per day.The subject receiving the composition can be exposed to the composition for several hours or minutes.The duration of exposure can depend on the frequency, amount or severity of infection.

[0164] The total daily amount of API formed into extemporaneous solution from the solid precursor can range from 0.01 to 1000 mg, depending on the nature of the OC. The actual dosage may vary depending on the specific composition administered, the mode of administration, and other factors known in the art.

[0165] The composition can be administered to any member of the respiratory tract, such as the respiratory epithelium, nasal cavity, nasal epithelium, pharynx, esophagus, larynx, epiglottis, trachea, carina, bronchi, bronchioles, or lungs. By administering the composition to the respiratory tract, any disease or disorder transmitted by a virus is treated or prevented.

[0166] In certain other embodiments, the compositions of the present invention can be used to disinfect, for example, entire rooms, facility medical equipment, and surgical instruments. Medical equipment supplies are often initially sterile, but may require additional or subsequent cleaning and disinfection or sterilization. In particular, it is particularly important to sterilize or disinfect reusable medical equipment before reuse using any known technique. The composition can be applied to the medical equipment using a disinfectant. For example, the composition can be applied by wiping or spreading the composition on the surface of the equipment, by spraying an aerosol or mist form of the composition on the equipment, by immersing the equipment in a container containing a certain amount of the composition, or by placing the equipment in a stream of the composition from a faucet or the like. Additionally or alternatively, medical equipment and surgical instruments can also be stored submerged in the composition and removed when used.

[0167] In summary, the disinfecting efficacy of the complete instant formulation of the present invention has been found to be greater than that of the components alone, as known in the art, and the difference in performance is readily observable over a wide range of concentrations.

[0168] Furthermore, because both oxidized chlorine species and glutaric acid are toxic in high concentrations, the prior art teaches against the use of these agents on skin or other tissues except in trace amounts. Thus, the present invention is surprisingly safe and effective when used in the controlled manner described above.

[0169] Some of the disclosed compositions contain 2% or more glutaric acid and have been proven safe and effective for treating skin and other tissues when combined with OCs. The OCs in these compositions have been found to have a modulating effect on glutaric acid.

[0170] This allows the composition to take advantage of the antiseptic properties of glutaric acid without harming tissue.

[0171] General procedure for preparation of a dry solid mixture of API-P and NaCl for inclusion in pre-calculated amounts in effervescent tablets.

[0172] Example 1. Preparation of OC powder and effervescent powder formulations

[0173] a) Preparation of powdered Ca(OCl)2 as part of a powder / effervescent powder / effervescent tablet / granule composition

[0174] I. 100 g of calcium hypochlorite granules were placed in a ceramic mortar (Porsgrunn Porselen, Norway). The granules were powdered several times using a mortar and pestle. The powder was sieved through ≦75 μm, ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm masks (Retsch, Haan, Germany). The powdered material was stored in various containers made of the following materials: PET, amber glass, and PE.

[0175] 100 g of calcium hypochlorite granules for dry grinding were sequentially placed in a Retsch mortar grinder (Haan, Germany). The final fineness of the powder was adjusted to ≦75 μm, ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm, respectively.

[0176] 100 g of dry-milled calcium hypochlorite granules were sequentially placed in a Retsch drum mill (Haan, Germany). The final fineness of the powder was adjusted to ≦75 μm, ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm, respectively.

[0177] b) Preparation of powdered sodium hydroxide (NaOH) as part of a powder / effervescent powder / effervescent tablet / granule composition

[0178] I. 100 g of sodium hydroxide pellets were added to a ceramic mortar (Porsgrunn Porselen, Norway). The granules were powdered several times using a mortar and pestle. The powder was sieved through ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm masks (Retsch, Haan, Germany). The powdered material was stored in various containers made of the following materials: PET, amber glass, and PE.

[0179] 100 g of sodium hydroxide pellets for dry grinding were sequentially placed in a Retsch mortar grinder (Haan, Germany). The final fineness of the powder was adjusted to ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm, respectively.

[0180] 100 g of dry-milled sodium hydroxide pellets were sequentially placed in a Retsch drum mill (Haan, Germany). The final fineness of the powder was adjusted to ≦150 μm, ≦250 μm, ≦355 μm, and ≦500 μm, respectively.

[0181] c) Preparation of a powder formulation with glutaric acid as the acid activator

[0182] 1.65 g of glutaric acid was mixed by volume with 0.33 g of powdered NaOH (particle size ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) and 102.0 mg of powdered Ca(OCl)2 (particle size ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) to provide a pre-calculated powder formulation that can be readily used to mix with water to release a stabilized hypochlorous acid solution.

[0183] d) Preparation of a powder formulation with glutaric acid as the acid activator

[0184] 1.48 g of succinic acid was mixed by volume with 0.35 g of powdered NaOH (particle size ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) and 102.8 mg of powdered Ca(OCl)2 (particle size ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) to provide a pre-calculated powder formulation that can be readily used to release a stabilized hypochlorous acid solution upon mixing with water.

[0185] e) Preparation of an effervescent powder formulation with glutaric acid as the acid activator

[0186] 3.30 g of glutaric acid was mixed by volume with 1.60 g of sodium bicarbonate and 220.9 mg of powdered Ca(OCl)2 (particle sizes ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) to obtain a pre-calculated effervescent powder formulation that could be easily mixed with water to release a stabilized hypochlorous acid solution. Effervescent time was determined by dissolving the effervescent powder in a beaker containing 100 ml of distilled water at 20°C. Effervescent time was measured chronometrically and was reached when the solution became clear and no particles were present.

[0187] f) Preparation of effervescent powder formulations with succinic acid as the acid activator

[0188] 2.95 g of succinic acid was mixed by volume with 1.61 g of sodium bicarbonate and 221.9 mg of powdered Ca(OCl)2 (particle sizes ≦150 μm, ≦250 μm, ≦355 μm, ≦500 μm) to obtain a pre-calculated effervescent powder formulation that could be easily mixed with water to release a stabilized hypochlorous acid solution. Effervescent time was determined by dissolving the effervescent powder in a beaker containing 100 ml of distilled water at 20°C. Effervescent time was measured chronometrically and was reached when the solution became clear and no particles were present.

[0189] Example 2. Determination of powder flowability

[0190] 100 g of powdered calcium hypochlorite with sizes ≦355 μm and ≦500 μm was added to a dry funnel (φ110 mm, diameter of outlet opening = 15 mm) with the bottom opening covered with a plastic cover. The bottom opening of the funnel was opened, and the time required for the entire sample to flow out was measured. The flow rates for particle sizes ≦355 μm and ≦500 μm were 35.7 g / s and 37.4 g / s, respectively. The angles of repose were calculated to be 29.5° and 26.8° for particle sizes ≦355 μm and ≦500 μm, respectively.

[0191] Example 3. Dissolution times of powdered calcium hypochlorite with particle sizes ≦250 μm, ≦355 μm, and ≦500 μm

[0192] 1.58±4 mg of powdered material of all sizes was dissolved in 100 mL of 0.25% wt. acetic acid / acetate buffer at 2500 rpm. Particle sizes ≤250 μm dissolved within 10 seconds, ≤355 μm within 15 seconds, and ≤500 μm within 1 minute 25 seconds.

[0193] II. 58±4 mg of powdered material of all sizes was dissolved in 100 mL of 3.0% wt. acetic acid / acetate buffer at 2500 rpm. Particle sizes ≦250 μm dissolved within 10 seconds, ≦355 μm within 25 seconds, and ≦500 μm within 25 seconds.

[0194] III. 570±40 mg of powdered material of all sizes was dissolved in 100 mL of 0.25% wt. acetic acid / acetate buffer. At 2500 rpm, particles ≦250 μm dissolved within 15 seconds, particles ≦355 μm dissolved within 45 seconds, and particles ≦500 μm dissolved within 1 minute 35 seconds.

[0195] IV. 570±40 mg of powdered material of all sizes was dissolved in 100 mL of 3.0% wt. acetic acid / acetate buffer. The procedure was repeated using 3.0% acetic acid / acetate buffer. At 2500 rpm, particles ≦250 μm dissolved within 15 seconds, ≦355 μm within 15 seconds, and ≦500 μm within 20 seconds.

[0196] Example 4. Preparation of effervescent tablets

[0197] a) Preparation of granulated material

[0198] Preferred molecular weights of carboxylic acids and bases useful in the present invention are: sodium hypochlorite (molecular weight: 74.44 g / mol), calcium hypochlorite (molecular weight: 142.98 g / mol), sodium chlorite (molecular weight: 90.44 g / mol), calcium chlorite (molecular weight: 157.89 g / mol), succinic acid (molecular weight: 60.05 g / mol), pyruvic acid (88.06 g / mol), glutaric acid (132.12 g / mol), citric acid (192.12 g / mol), malic acid (134.09 g / mol), oxaloacetic acid (132.07 g / mol), tartaric acid (150.09 g / mol), adipic acid (146.14 g / mol), or fumaric acid (116.07 g / mol).

[0199] The processing temperature of 55°C is imposed by the physicochemical constraints of the ingredients (e.g., dimethyl polyethylene glycol (PEG6000-di-Me, dimethyl ether) with an average molecular weight of 6000 D and sodium bicarbonate). The effervescent powder contains 100 g of a stoichiometric effervescent system formed from succinic anhydride (43.2%) / sodium bicarbonate (56.8%) as a medium-coarse or very fine powder, to which micronized PEG6000-di-Me is added as a molten material. After mixing for 10 minutes at 28 rpm in a Turbula mixer (T2A, Basel, Switzerland), the effervescent mixture is transferred to a vertical fluidized-bed dryer (Uni-Glatt, Binzen, Germany), initially adjusted to 55°C and 123 m / hr. Granulated materials are obtained by fusion of particles by melting PEG6000-di-Me for 5, 15, or 30 minutes. After cooling for 30 minutes at ambient temperature and 30% relative humidity (RH), the granules were sieved in an oscillating granulator (Erweka FGS, Frankfurt, Germany) set at medium speed (II) and fitted with an 800 μm sieve.

[0200] Measurement of physical properties of granulated foaming materials

[0201] Granule flowability and density Granule flowability and density were measured using accurately weighed 100 g samples. Granule flowability was measured using a flow meter consisting of a standardized funnel and a chronometer. Tap density was determined using a volumetric meter (Jel / Stav 2003A, Ludwigshafen, Germany).

[0202] Granule size distribution: Granule size distribution was analyzed using a vibrating Siever Retsch (Haan, Germany) (amplitude 1.5; 10 min) equipped with 100 g of granules and a series of European Pharmacopoeia (3rd edition) sieves (710, 500, 355, 250, 180, 125 μm). The mean granule size was determined graphically using a log-normal distribution chart.

[0203] Foaming time Foaming time was determined using 3 g of granules accurately weighed into a beaker containing 200 ml of distilled water at 20° C. Foaming time was measured chronometrically; the end of foaming was reached when the solution became clear and no particles were present.

[0204] Tableting: We investigated the granule properties that would form tablets with a crush resistance between 70N and 120N when compressed in a single punch tablet press. After mixing 700 g of effervescent granules with sodium benzoate or siliconized sodium benzoate in a Turbula mixer at 28 rpm for 10 minutes, we obtained a granule mixture ready for compression into tablets. Siliconized sodium benzoate was prepared by mixing 600 g of ultrafine sodium benzoate powder with 40 g of Silbione (silicone) in a Kenwood planetary mixer at low speed (II) for 30 minutes. The mixture was sieved using a 125 μm sieve. Silbione is an oil-based lubricant that can act to enhance the effectiveness of sodium benzoate. Tablets were manufactured using a single-punch tablet press (Frogerais OA, Evry Lisses, France) equipped with 24 mm diameter chrome-plated punches in a climate-controlled room at 30% RH (relative humidity) and 22°C (relative humidity). Tablet weight, crush resistance (Erweka TBH 28, Heusenstamm, Germany), friability (Pharma Test PTF 1E, Haiberg, Germany), and foaming time were evaluated using the European Pharmacopoeia (10th edition) methods. Tablet moisture content, carbon dioxide content, solution pH, and tablet and solution appearance were evaluated using the same methods as for granules. Adhesion to punch faces and die walls, capping, and sliding friction at the die walls were also evaluated visually. All formulations were compressed under constant conditions (depth of the lower punch in the die was 6.98 mm, and the distance that the upper punch penetrated into the die was 6.04 mm) to select tablets with a weight of more than 2 g and a crushing resistance between 70 N and 120 N without any processing problems during compression. physicochemical properties

[0205] The pH was determined using an Aquadata APH 1000 pH meter. Carbon dioxide content. The carbon dioxide content of an accurately weighed 3 g of granules or one tablet in 100 ml of dilute sulfuric acid (R) was determined using a Mettler PG 503S precision balance (Viroflay, France) (18, 19). Results were expressed as the loss of sample weight at the end of effervescence (mg of CO2 per gram of ES).

[0206] Using this procedure, tablets typically have a moisture content of less than 0.1, an average granule size of 510μ, a CO2 content of 300mg CO2 per gram of granule, an effervescence time of 70 seconds min, a pH of the resulting solution of 5.6, and a typical dissolution volume of 5mL per tablet in a pharmaceutically acceptable carrier in water or ambient environment.

[0207] Example 5 In vitro anti-biofilm efficacy of three exemplary different test solutions of HOCl and succinic acid.

[0208] Three different test solutions were generated from the effervescent tablet: All three test solutions are generated from the effervescent tablet.

[0209] Experimental setup

[0210] Test organisms: Pseudomonas aeruginosa or Staphylococcus aureus wild-type strains. Biofilm type: 48- or 24-hour-old biofilms grown on semipermeable membranes placed on solidified medium supplemented with 0.5% glucose. For 48-hour-old biofilms, the membranes with biofilms were transferred onto new plates after 24 hours.

[0211] Initial viable cell count: 5 x 10 9 colony forming units (CFU)

[0212] Treatment method: The membrane containing the biofilm was transferred to a new plate. Eight to ten layers of sterile gauze were placed on the second membrane, and 1 ml of antibacterial solution was pipetted onto the gauze layer. Treatment was carried out at room temperature for 2 to 3 hours or 4 to 6 hours. For the 4 to 6 hour treatment, the gauze layer was replaced with a new one containing 1 ml of sample solution 2 or 3 hours after the start of treatment.

[0213] Evaluation method: The gauze layer was discarded and each membrane with biofilm was transferred to a 15 ml tube containing 5 ml of 0.9% NaCl, vortexed for 10 seconds, sonicated in an ultrasonic bath for 10 minutes, and vortexed again for 10 seconds. Ten-fold serial dilutions were made and 10 ul of each dilution was spot-plated onto LB plates for viable CFU counts.

[0214] Results and Conclusions

[0215] Figure 2 shows the results obtained using the sample solutions. Increasing the acid activator concentration in the 200 ppm HOCl solution from 0.25% to 1% and 2% gradually increased the killing of Staphylococcus aureus biofilms. 1% glutaric acid alone had only a slight effect on the biofilms. The three test solutions were compared with four different competing wound healing products on the market, and all of these products showed only a slight effect on Staphylococcus aureus biofilms. They showed even stronger effects on biofilms derived from Pseudomonas aeruginosa. We conclude that hypochlorous acid and succinic acid at pH 4-7 act synergistically and efficiently at concentrations shown to be safe in other studies.

[0216] Example 6: In vivo toxicity studies

[0217] Example 6.1: 7-day inhalation toxicity study in rats.

[0218] A 7-day inhalation toxicity study in rats was performed as described by Kogel et al. in Food Chem Toxicol. 2014 Jun;68:204-17. The rat inhalation study was conducted in accordance with the Organization for Economic Cooperation and Development (OECD) guidelines. Test solutions were generated from effervescent tablets. Sprague-Dawley rats were exposed to either filtered fresh air (sham) or test solutions as a reference. Animal care and use followed the American Association for Laboratory Animal Science (1996) guidelines. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC). Histopathological evaluation was performed at defined anatomical sites in the nose and left lung according to a defined grading system. Free lung cells were determined in bronchoalveolar lavage fluid by flow cytometry, and inflammatory mediators were measured by multianalyte profiling (MAP). For the systems toxicology approach, RNA samples were obtained from specific sites in the airway: the respiratory nasal epithelium (RNE) and lung. For lung RNA isolation, respiratory epithelium from the main bronchi and lung parenchyma was separated by laser capture microdissection (LCM), further processed, and analyzed on whole-genome Affymetrix microarrays (GeneChip® Rat Genome 230 2.0 Array). No major perturbations related to inflammation, cellular stress, or cell proliferation were observed in the bronchi or lung parenchyma.

[0219] Example 7: Treatment of Mastitis

[0220] For applications where the color indicator in step 4 can add information in the treatment procedure, for example in or for indication of oxidative activity of the API, a compartment containing the ROD is included in the procedure.

[0221] Example 8: Clinical Antiviral Therapy

[0222] Five milliliters of test solution, generated from an effervescent tablet, was loaded into the medicine cup of a Gima Aerosol Corsia Nebulizer. The patient's mouth was attached to the hose and face mask attached to the nebulizer, and the nebulizer was started. After 10 to 15 minutes of breathing, the fluid was exhausted and the nebulizer was stopped. The patient was monitored for several hours to ensure no adverse reactions occurred. The patient's mucous membranes and cilia were examined for potential side effects.

[0223] INCORPORATION BY REFERENCE All references and citations made throughout this disclosure to other documents, such as patents, patent applications, patent publications, journals, books, articles, web content, etc., are incorporated herein by reference in their entirety for all purposes.

[0224] equivalent The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics, and the foregoing embodiments are therefore to be considered in all respects as illustrative and not restrictive of the invention described herein.

Claims

1. An antimicrobial composition comprising a solid chlorine oxide species salt of the following formula: Mn + [C1(O) x ] a w- (Here, M is an alkali metal, alkaline earth metal, or transition metal ion, n, a, and w are each independently 1 or 2, and x is an integer from 1 to 4); Acid activators having the following formula: RN(COOH) m (wherein RN is a saturated or unsaturated organic moiety containing 1 to 25 carbon atoms, which may be substituted with oxygen or hydrogen as needed to form a functional group that cannot be oxidized by chlorinated species, and m is an integer from 0 to 10) or a salt thereof; Pharmacopoecially acceptable diluents, adjuvants, or carriers; and A base activator having the following formula that can react with the acid activator to produce carbon dioxide: (M' z+ ) s ((H) t CO 3 ) u (Here, z is 1 or 2, s is 1 or 2, t is 0 or 1, u is 1 or 2, and t + s + z is an integer from 1 to 5.) M' is an alkali metal, alkaline earth metal, or transition metal. An antimicrobial composition containing the above.

2. The composition according to claim 1, wherein the chlorine oxide salt comprises an alkali metal salt or alkaline earth metal salt of hypochlorous acid.

3. The composition according to claim 1, wherein the chlorine oxide salt comprises an alkali metal salt or alkaline earth metal salt of chlorite.

4. The composition according to claim 1, wherein the acid activator is selected from the group comprising monovalent acid, divalent acid, trivalent acid, propionic acid, lactic acid, succinic acid, glutaric acid, pyruvic acid, citric acid, malic acid, oxaloacetate, tartaric acid, adipic acid, fumaric acid, heptaneoic acid, octanedioic acid, diacid, and derivatives thereof.

5. The composition according to claim 1, wherein the base activator is selected from the group comprising sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate.

6. The composition according to claim 1, further comprising a water-soluble polyol binder.

7. The composition according to claim 1, further comprising a water-soluble polyol binder selected from the group comprising monosaccharides, disaccharides, polysaccharides, or monohydroxy acids or polyhydroxy acids.

8. The composition according to claim 1, further comprising a lubricant.

9. The composition according to claim 1, further comprising a lubricant selected from the group consisting of dextrose, lactose, sorbitol, ascorbic acid, sorbitol, mannitol, sodium benzoate, potassium sorbate, polyethylene glycol or its derivatives, succinic acid, adipic acid, glutaric acid, or clavulanic acid.

10. The composition according to claim 1, wherein the composition has a weight osmolality in the range of about 0.1 mOsm to about 500 mOsm.

11. The composition according to claim 1, wherein the composition has a pH between pH 4 and pH 8.

12. The composition according to claim 1, further comprising a viscosity enhancer.

13. The composition according to claim 12, wherein the viscosity enhancer comprises a water-soluble gelling agent.

14. The composition according to claim 13, wherein the water-soluble gelling agent is selected from the group consisting of polyacrylic acid, polyethylene glycol, poly(acrylic acid)-acrylamide alkylpropanesulfonic acid copolymer, phosphinopolycarboxylic acid, poly(acrylic acid)-acrylamide alkylpropane, and sulfonic acid-sulfonated styrene terpolymer.

15. The composition according to claim 1, further comprising a colored dye that provides a visual indication of the presence of a chlorine oxide compound.

16. The composition according to claim 15, wherein the aforementioned dye comprises a redox dye.

17. The composition according to claim 15, wherein the color and color intensity of the dye depend on the oxidation state of the chlorine oxide compound.

18. The composition according to claim 1, wherein the solid component is contained within an effervescent powder, granules, or tablet.

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