Compositions and methods for treating and preventing pathogens

Abstract

The present invention provides a novel storable and stable disinfectant composition comprising a solid precursor in an oxidation state using chlorine. The components in the device do not have stability issues because these solid precursors are immediately dissolved in water or pharmaceutically acceptable diluents, adjuvants or carriers, and in combination with activators such as adipic acid or succinic acid or salts thereof, optionally in combination with viscosity enhancers, optionally in combination with dyes. The main product according to the invention is produced by dissolving an effervescent material in the form of tablets or microparticles, the resulting solution produced by dissolving the effervescent material being a useful disinfectant for the treatment of a broad spectrum of pathogenic bacteria and / or viruses, fungi or parasitic pathogens, representative of microorganisms.

Description

Technical Field

[0001] The present invention generally relates to compositions for treating and preventing pathogens, the compositions comprising combinations of one or more solid precursors of oxidation states of chlorine. Background Art

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

[0003] Free-living microorganisms such as bacteria or fungi have the potential to grow on surfaces or within host organisms, causing infection of the host organism, which can become pathological and can lead to disease or death.

[0004] Infectious diseases are a leading cause of death worldwide and cause more than 13 million deaths each year, including nearly two-thirds of all child mortalities. In addition, antibiotic resistance is increasing and contributing to the incidence of multiple human diseases, including pneumonia, tuberculosis, and cholera. Of particular concern is that many human pathogens have developed multi-resistance to conventional antibiotics. The introduction of new, more effective derivatives of existing antibiotics has only provided a temporary solution because existing resistance mechanisms rapidly adapt to the new derivatives (Hoiby et al., Int. J. Antimicrob. Agents 2010). While resistant Gram-positive bacteria pose a major threat, the emergence of multi-drug resistant (MDR) strains of common Gram-negative pathogens such as Escherichia coli is of particular concern. Pan-resistance or extreme resistance are currently common terms used to describe clinically important isolates of Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterobacteriaceae that are resistant to almost all antibiotics.

[0005] Viruses are also an important issue in infectious epidemiology. Severe viral outbreaks are becoming increasingly common, many of which are zoonotic. For example, the SARS (severe acute respiratory syndrome) and MERS (Middle East respiratory syndrome) outbreaks in the early to mid-2000s, the 2009 H1N1 pandemic, and the subsequent 2020 SARS CoV-2 pandemic have focused attention on both treating and preventing the spread of these viral pathogens.

[0006] Many viruses that infect the respiratory tract are transmitted by droplet infection. In this case, respiratory droplets containing the virus are expelled by an infected person and acquired by others through direct contact or contact with surfaces onto which the droplets have fallen. Typically, infection occurs by binding of the virus to receptors on mucosal or epithelial cells, followed by entry into the nose, eyes, ears, or mouth. Additionally, other viruses are transmitted via virus-containing aerosol particles or by airborne transmission. In both cases, the virus can survive for hours to days after being expressed from an infected individual.

[0007] Infections can be caused by microorganisms such as the viruses, bacteria, fungi, spores, parasites, and combinations thereof described herein. The viruses 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 causing an outbreak of severe acute respiratory syndrome (SARS), and the MERS-CoV virus, identified in 2012 as causing 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, for example, infections caused by one or more of the following: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Acinetobacter baumannii, Escherichia coli, Staphylococcus aureus, Bacillus atrophaeus, Streptococcus pyogenes, Samonella choleraesuis, Shigella dysenteriae, Mycobaterium tuberculosis, etc. Further, problematic fungi and yeasts can include, for example, one or more of the following: Candida albicans, Bacillus subtilis, Trichophyton mentagrophytes, and Bacillus atrophaeus.

[0009] Conventional compositions and methods for disinfecting inanimate surfaces or contaminated epithelial cells are not sufficient to inactivate all of these infectious agents. The current forms of conventional disinfectant compositions and methods may require long and impractical exposure times, or cannot be used with dangerous or corrosive solutions or vapors for expensive instruments or living tissues, and thus cannot provide a ready solution for the growing health risks of drug and disinfectant-resistant agents.

[0010] It is clear that there is a significant unmet medical need for new methods of treating resistant microorganisms and viruses, specifically methods that are effective against the critical stages by which microorganisms enter mammalian cells through biological mechanisms and are also effective outside of mammalian biology. Summary of the Invention

[0011] The present invention generally relates to compositions for eliminating pathogenic viruses, bacteria, and other infectious diseases. In a preferred embodiment, the present invention comprises a solid precursor of one or more oxidation states of chlorine, an acid activator, and its salts and a base activator. In certain embodiments, these acid activators and base activators are solids. The compositions of the present invention may also comprise a viscosity enhancer and / or a dye or a redox-activated dye. The compositions of the present invention are formulated as effervescent powders, tablets, or microparticles. When dissolved in water, the resulting solution provides a disinfectant for treating a broad spectrum of bacterial, viral, fungal, and parasitic pathogens.

[0012] The compositions of the present invention are provided in the form of effervescent powders, tablets, or microparticles and are soluble in water or a pharmaceutically acceptable diluent, adjuvant, or carrier. The oxidized chlorine species are combined with the acid and base activators in the tablet, and the acid activator synergistically acts with the oxidation state of chlorine. The acid activator may preferably be a carboxylic acid and / or its salts, preferably having a melting point above 20 degrees Celsius. Optionally, the tablets as described herein may comprise a viscosity enhancer and / or optionally a dye. The compositions of the present invention can be used as disinfectants for treating a broad spectrum of bacteria, viruses, and / or other pathogens.

[0013] In a specific aspect, the present invention provides an antimicrobial composition comprising a solid chlorine oxide species salt, an acid activator, a base activator, and a pharmaceutically acceptable diluent, adjuvant, or carrier. The solid chlorine oxide species salt is based on 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, including 1 and 4. The acid activator is based on the formula RN(COOH) m ​or a salt thereof, wherein RN is a saturated or unsaturated organic moiety containing 1-25 carbon atoms, which organic moiety is optionally substituted with oxygen or hydrogen to form a functional group that cannot be oxidized by a chlorinated species, and m is an integer from 0 to 10. The base activator is capable of reacting with the acid activator to produce carbon dioxide, and is based on the formula (M n+ ) s ((H) t CO3) u , wherein 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 solid chlorine oxide species salt according to the following 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; an acid activator having the following formula: RN(COOH) m or a salt thereof, wherein RN is a saturated or unsaturated organic moiety containing 1-25 carbon atoms, which organic moiety is optionally substituted with oxygen or hydrogen to form a functional group that cannot be oxidized by a chlorinated species, and m is an integer from 0 to 10; a pharmaceutically acceptable diluent, adjuvant or carrier; and a base activator capable of reacting with the acid activator to produce carbon dioxide, the base activator having the following formula: (M' z+ ) s ((H) t CO3) u , wherein 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.

[0015] In some embodiments, the chlorine oxide salt comprises an alkali metal or alkaline earth metal salt of hypochlorous acid. In other embodiments, the chlorine oxide salt comprises an alkali metal or alkaline earth metal salt of chlorous acid. In certain embodiments, the acid activator is selected from the group consisting of: monocarboxylic acids, dicarboxylic acids, tricarboxylic acids, diacids, propionic acid, lactic acid, succinic acid, glutaric acid, pyruvic acid, citric acid, malic acid, oxaloacetic acid, tartaric acid, adipic acid, fumaric acid, pimelic acid, suberic acid, diacids 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 composition of the present invention further comprises a water-soluble polyol binder. In some embodiments, the water-soluble polyol binder is selected from the group consisting of monosaccharides, disaccharides, polysaccharides, or mono-hydroxy acids or poly-hydroxy acids.

[0018] The composition of the present invention may further comprise a lubricant. Preferred lubricants include glucose, lactose, sorbitol, ascorbic acid, sorbitol, mannitol, sodium benzoate, potassium sorbate, polyethylene glycol and its derivatives, succinic acid, adipic acid, glutaric acid, and clavulanic acid.

[0019] In some embodiments, the composition of the present invention is formulated as an aqueous solution, gel, cream, ointment, or oil. In some embodiments, the osmotic pressure of the dissolved antimicrobial composition ranges from about 0.1 mOsm to about 500 mOsm. In some embodiments, the pH of the dissolved composition is between 4 and 8.

[0020] The composition of the present invention may further comprise a viscosity enhancer. In some embodiments, the viscosity enhancer 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)-acrylamidoalkylpropanesulfonic acid copolymer, phosphonopolycarboxylic acid, and poly(acrylic acid)-acrylamidoalkylpropane and sulfonic acid-sulfonated styrene terpolymer.

[0021] The composition of the present invention may additionally comprise a dye, which includes a color providing a visual indication of the presence of the chlorine oxide 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 chlorine oxide compound.

[0022] In some embodiments, the solid components of the antimicrobial composition are contained within an effervescent powder, microparticles, or tablets.

[0023] In certain aspects, the present invention provides an antimicrobial composition that can be used for antiviral applications and antimicrobial applications.

[0024] The formulation of the present invention can be used for inhalation therapy using an asthma inhaler, nebulizer, or vaporizer to combat viral infections in the upper airway of mammals.

[0025] The formulation of the present invention can be used for the prevention or healing of skin or wounds.

[0026] The formulation of the present invention can be used to treat mastitis or any other infectious disease in animals or agricultural breeding.

[0027] The formulation of the present invention can be used to treat any infectious disease in aquaculture.

[0028] The formulations of the present invention can be used to treat any chemical or biological warfare agent.

[0029] In a specific aspect, the present invention provides a method for preparing effervescent powders, microparticles, and tablets from an antimicrobial composition. In some embodiments, powders, microparticles, and tablets are prepared from the antimicrobial composition using wet granulation, fluidized bed dryers, or vacuum granulation machines.

[0030] In some embodiments, the present invention provides a method for neutralizing chemical or biological warfare agents, using the step of dissolving an effervescent material in a liquid phase and generating vapor or aerosol in the area or location of interest. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 is a schematic diagram showing an exemplary effervescent tablet for dispensing a disinfectant composition according to an embodiment of the present invention.

[0032] Figure 1 Shows an effervescent tablet prepared from an effervescent powder or microparticle, the effervescent tablet comprising 1. chlorine oxide species or its salts, 2. an acid activator comprising at least one carboxylic acid functional group, 3. salts of molecules optionally comprising at least one carboxylic acid functional group, 4. a base activator, 5. an optional binder, 6. an optional lubricant, 7. an optional viscosity enhancer, 8. an optional dye, and 9. an optional sodium chloride to obtain an osmotic pressure close to that of blood. These tablets can be prepared from effervescent powders or microparticles using wet granulation, fluidized bed dryers, vacuum granulation machines, or any other method suitable for tablet production using the materials according to the present invention. DETAILED DESCRIPTION

[0033] The present invention generally relates to compositions comprising combinations of solid and liquid precursors of chlorine in its oxidized states with activators, such as glutaric acid or its salts, and one or more additional components. Such compositions are used as antimicrobials and disinfectants for treating a broad spectrum of bacterial and / or viral pathogens on various biological and non-biological surfaces and environments.

[0034] The compositions of the present invention can be formulated as solids, such as in the form of effervescent powders, tablets, or microparticles. This addresses the problem of shortened shelf life of hypochlorous acid or chlorine dioxide solutions described in the prior art. More specifically, ready-to-use formulations that directly generate chlorine oxide species from solid precursor API-P can be carried out in effervescent tablets at the point of use. The effervescent powders, tablets, or microparticles according to the present invention are for dispensing, but show long-term stable storage of solid compositions consistent with the present invention. In an example, solid precursors of chlorine in its oxidized state and glutaric acid or its salts, a viscosity enhancer, and a dye are combined and then such components are combined at the desired time and location of use to prepare the composition.

[0035] As background, chlorine oxides or oxychlorines (also referred to herein as “OC”) encompass a large class of chemical species and are often found in nature as well as in mammalian biological systems. Chlorine oxides can also exist in the form of neutral compounds or ions, namely the so-called oxyanions. There are several oxyanions of chlorine, where the oxyanions can exhibit oxidation states of +1, +3, +5 or +7 in correspondence with the anions hypochlorite (ClO - ), chlorite (ClO2 - ), chlorate (ClO3 - ) or perchlorate (ClO4 - ). The standard reduction potential of hypochlorous acid (HOCl) at low pH is +1.49, and that of chlorous acid (HClO2) is 1.64, while at alkaline pH, the standard reduction potentials are +0.89 and +0.78 respectively. At a pH of 5 to 7, the reduction potential is higher than +1.

[0036] Therefore, hypochlorite and chlorite are generally the most useful oxidation states, having the potential to kill microorganisms and parasites at low pH. Specifically, the chloride ion Cl - is in the most stable oxidation state and is non-reactive and not an effective disinfectant. Chlorate and perchlorate in the oxidation states +5 and +7 are more reactive than the lower oxidation states and may be more difficult to handle.

[0037] The hypochlorite ion has the chemical formula ClO - , where chlorine (Cl) is in the oxidation state +1, which is a potentially unstable oxidation state since the low-energy oxidation state of Cl is -1. Both the hypochlorite ion and the chlorite ion combine with many cations to form hypochlorites and chlorites as salts of these chlorine oxides. Common examples include sodium hypochlorite (household bleach) and calcium hypochlorite, which is the main active ingredient of commercial products including bleaching powder, chlorinated powder or chlorinated lime, and is commonly used in water treatment (e.g., swimming pools, etc.). Chlorite and hypochlorite ions are also referred to herein as “primary chlorine oxides” and are useful in various situations. Sodium chlorite and hypochlorite are strong oxidants and have been used in water purification, disinfection, and the bleaching and deodorization of animal products.

[0038] Since sodium hypochlorite produces highly toxic chlorine gas under acidic conditions, commercially available household aqueous solutions are strongly alkaline solutions with the pH adjusted using sodium hydroxide.

[0039] Chlorites and hypochlorites can also be used to treat various diseases or conditions. For example, chlorites and hypochlorites can be used to treat infections, as described in U.S. Patent Nos. 4,725,437 and 4,851,222. Chlorites and hypochlorites can also be used to treat HIV, recurrent prostate cancer, cystitis, and chronic active hepatitis C (see, e.g., McGrath et al., Development of WF10, a novel macrophage-regulating agent, Curr. Opin. Investig. Drugs, 3(3):365-73 (March 2002)). Chlorites and hypochlorites have also been described for use in treating oral or periodontal diseases or conditions such as gingival inflammation (see, e.g., U.S. Patent No. 6,350,438).

[0040] Hypochlorous acid (HOCl) is widely used as a broad-spectrum household and industrial disinfectant and deodorizer. Hypochlorous acid is a weak acid and is known to rapidly inactivate bacteria, algae, fungi, and other organic matter, making it an effective agent against a wide range of microorganisms. Additionally, since hypochlorous acid is a weak acid and since humans naturally produce certain compounds that allow them to tolerate hypochlorous acid, hypochlorous acid is generally harmless to humans. Due to the combination of the biocidal properties and safety characteristics of hypochlorous acid, hypochlorous acid has been found to have many beneficial uses in many different industries, such as medical, food service, food retail, agriculture, wound care, laboratories, hotels, dentistry, or the floral industry.

[0041] Hypochlorous acid is formed when chlorine dissolves in water. Specifically, the acidification of hypochlorite produces hypochlorous acid, in which the chlorine atom is in the +1 oxidation state. Hypochlorous acid is in equilibrium with chlorine gas in the presence of Cl - and can escape from the solution. This equilibrium is pH-dependent, as shown by the following equation (Equation 1):

[0042]

[0043] Referring to the above equation (Equation 1), high pH drives the reaction to the right, promoting the disproportionation of chlorine into chloride and hypochlorite, while low pH drives the reaction to the left, promoting the release of chlorine gas (Cl2).

[0044] Chlorine gas (Cl2) is a toxic and potent oxidizing agent. An adverse property of chlorine gas is its tendency to insert Cl into hydrocarbons, forming mono- or polyhalogenated-alkanes and other chlorinated hydrocarbons in radical reactions (see Barhorst and Kubiak, Environ. Sci. Pollut. Res. (2009) 16, pp. 582 - 589). This is potentially dangerous because halogenated hydrocarbons are pollutants and may be considered carcinogenic. Therefore, chlorine gas is not a desired compound for use at higher concentrations in nature or in medical applications. At lower concentrations, chlorine contributes to the powerful disinfectant properties of hypochlorous acid in equilibrium, while having a lower reasonable risk of medical side effects. However, as long as the pH of the hypochlorous acid solution is controlled and maintained within the range of 4 - 8, the amount of Cl2 can be negligible.

[0045] The antimicrobial action of hypochlorous acid is highly pH-dependent and is highest at pH values between 4 and 5.5, but is also highly active at physiological pH (7.1 - 7.5). This means that hypochlorite solutions should ideally be buffered with a weak organic acid and its corresponding metal salt to have maximum antimicrobial activity.

[0046] Generally, formulations containing chlorine oxide are effective antimicrobial agents with proven antimicrobial and antiprotozoal properties and can be used in disinfection techniques involving human and animal health. However, the formulations in the prior art have drawbacks. For example, the weak acid HOCl is unstable and impure when produced under conventional conditions. Therefore, a more controlled and direct preparation method is needed that can provide chlorine oxide with stability allowing for expected short-term use on-site.

[0047] The alkalinity of household hypochlorite bleach can be around pH 12, while HOCl solutions are weakly acidic with a pKa value of about 7. Therefore, HOCl formulations with a pH range around physiological pH 7 are more suitable for medical applications because bleach is harmful to the user and its application surfaces. Therefore, it is of utmost importance to keep the pH within the range of 4 to 9 to avoid damage to biological tissues or delicate surfaces.

[0048] In therapeutic techniques, an active pharmaceutical ingredient (also referred to herein as "API") is understood to be one or more chemical components that produce the corresponding therapeutic effect. In the prior art where hypochlorous acid is the sole active API, a weakness of the formulation is the lack of a biocompatible activator and pH stabilizer that simultaneously acts as both a buffer and an antimicrobial agent and preferably also synergistically acts with the API.

[0049] Another weakness of the prior art is that, in these cases, many aqueous solutions are produced by electrolysis of isotonic 0.9% NaCl / H2O (physiological saline) and have no buffering capacity because there is no mixture of weak biocompatible organic acids and their salts. Further, electrolysis is a rather complex and sometimes inconvenient method for preparing antimicrobial solutions (i.e., in war zones, tourist resorts or disaster or pandemic areas).

[0050] Another weakness of prior art chlorine dioxide-based formulations is lack of stability, which is based at least in part on the fact that chlorine dioxide degrades and decomposes gradually within the first few months of storage unless stored at low temperature and in the absence of light and oxygen. Thus, pharmaceutical shelf-life stability of chlorine oxide solutions cannot be achieved under ambient conditions.

[0051] Yet another weakness of prior art chlorine dioxide-based formulations is lack of a controlled ionic strength suitable for use with a given formulation. Administering any given formulation comprising chlorine dioxide to a mammal ideally requires an ionic strength of 300 mOsm to be isotonic with body fluids, equivalent to a concentration of approximately 150 mM NaCl.

[0052] Hypochlorous acid may be an attractive compound to include in an antimicrobial solution because neutrophils in the body naturally produce hypochlorous acid in its pure form. Further, circulating monocytes, tissue-resident macrophages, and microglia in mammals also produce hypochlorous acid to inactivate pathogens within phagocytic vesicles and in the extracellular space around phagocytes in tissues. Thus, hypochlorous acid is a natural biocompound produced by mammals at relatively high concentrations. Activated neutrophils can mobilize their primary granule enzymes to the cell surface. For example, as described by Hirche et al. in The Journal of Immunology (J. Immunol.) (2005); 174:1557-1565, the enzyme myeloperoxidase (MPO) in activated neutrophils uses hydrogen peroxide (H2O2) catalyzed by NADPH oxidase to produce HOCl.

[0053] Given the large number of neutrophils that accumulate in inflamed tissues such as the lungs of patients with acute respiratory distress syndrome, the concentration of HOCl available for clinical use has significant physiological relevance. It is well known in the prior art that 5x10 6 activated neutrophils can produce up to 100 μM HOCl within 2 hours. Thus, the mammalian biological system can handle concentrations of at least 100 μM HOCl without producing pathological consequences.

[0054] Another chlorine oxide used as an API in antimicrobial formulations is chlorine dioxide, where the chlorine atom is in the oxidation state +3. The main reaction of sodium chlorite is to produce chlorine dioxide, as shown in the following equation (Equation 2):

[0055]

[0056] Referring to the above equation (Equation 2), HOR is usually an inorganic acid such as HCl or citric acid, because a proton source is needed to first convert sodium chlorite to chlorous acid and then to chlorine dioxide, which is a highly water-soluble gas at room temperature.

[0057] An advantage of chlorine dioxide is that it does not produce chlorine gas Cl2, which is known to react with chlorinated hydrocarbons such as trihalo-methanes, which are toxic environmental pollutants. Another advantage of chlorine dioxide is that its activity as a disinfectant or the stability of its aqueous solution is not pH-dependent.

[0058] Chlorine dioxide is produced from sodium chlorite and is FDA-approved for disinfecting water and for washing fruits, vegetables, and poultry under certain conditions. Sodium chlorite, NaClO2, is a solid precursor of chlorine dioxide and is sometimes used in combination with zinc chloride. It has been found that sodium chlorite can also be used as a component of therapeutic mouthwashes, mouth rinses, toothpastes and gels, oral sprays, as a preservative in eye drops, and as a component in a contact lens cleaning solution marketed under the trade name Purite.

[0059] Chlorine dioxide is also used for the bleaching and delignification of textiles, pulp, and paper. After conversion to chlorine dioxide, sodium chlorite is also used for the disinfection of municipal water treatment plants. Chlorine dioxide is used for the sanitization of hard surfaces that come into contact with food and for washing or rinsing a variety of foods, including red meat, poultry, seafood, fruits, and vegetables. Because chlorine oxide compounds are unstable even when properly prepared, there is no measurable residue on food after disinfection. Chlorine dioxide is also used as a teat dip to control mastitis in dairy cows.

[0060] The U.S. Army Natick Soldier Research, Development and Engineering Center has produced a portable "power-free" method for generating chlorine dioxide, i.e., ClO2 gas, which is a highly effective biocide that can be used to eliminate contaminants ranging from benign microorganisms and food pathogens to Category A bioterror agents. In the weeks following the 9 / 11 attacks, when anthrax was mailed to government officials in letters, hazardous materials teams used ClO2 to decontaminate the Hart Senate Office Building and the Brentwood Postal Facility.

[0061] In dealing with the COVID-19 pandemic, the U.S. Environmental Protection Agency (EPA) has included ClO2 as an agent meeting its standards for environmental measures against pathogenic coronaviruses (see U.S. EPA, OCSPP (March 13, 2020) "List N: Disinfectants for Use against SARS-CoV-2". U.S. EPA retrieved March 28, 2020, title "How we know disinfectants should kill the COVID-19 coronavirus", Chemical & Engineering News, retrieved March 31, 2020).

[0062] However, in these cases, such formulations also suffer from the stability problems described above because these techniques also lack biocompatible activators and stabilizers for the precursors of chlorine oxides that act synergistically in the antimicrobial action of the final formulation.

[0063] In summary, in the prior art, the main challenge for the medical use of chlorine solutions with an oxidation state higher than -1 is stability, or lack thereof, because these chemical species are in a higher energy state and tend to return to chloride ions, Cl. -And will decompose in solution at ambient temperature. This hinders the shelf-life stability of the chlorine oxides required for pharmaceutical formulations and medical devices under ambient conditions. Thus, it is difficult for chlorine oxide solutions to achieve an appropriate shelf-life for medical devices and pharmaceuticals. This inherent limitation of all chlorine oxides restricts transportation and storage, especially in hot regions with variable temperature, humidity, and atmospheric gases.

[0064] Effervescent powders, tablets, or microparticles contain components of an acid and a base, which are typically in solid form and produce carbon dioxide when mixed with water or a pharmaceutically acceptable carrier. Typical non-limiting examples of acids that can be used in this reaction are monobasic, dibasic, or tribasic 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 pimelic acid or suberic acid or higher dibasic acids or their derivatives. Citric acid imparts a citrus-like taste to the product. However, according to the present invention, acids that cannot be oxidized by chlorine species are preferred acids, such as, but not limited to, succinic acid, glutaric acid, pyruvic acid, oxaloacetic acid, and adipic acid, as exemplified.

[0065] Typical bases used in the effervescent reaction are sodium bicarbonate, potassium bicarbonate, sodium carbonate, and potassium carbonate. Sodium bicarbonate is very common in effervescent formulations and produces a clear solution after tablet disintegration. These two types of carbonates are mainly used as desiccants to form a dry product.

[0066] Water-soluble binders are usually necessary in effervescent tablets to achieve a tablet hardness that can be handled. Non-limiting examples of binders are monosaccharides, disaccharides, polysaccharides, or mono- or polyhydroxy acids. Particularly preferred binders are glucose, lactose, sorbitol, ascorbic acid, sorbitol, or mannitol. The above-mentioned carboxylic acids can also be used as lubricants. The ideal amount of binder is an amount that makes the tablet hard enough to be easily handled, soft enough to be easily disintegrated, and dry enough to be stable. The harder the tablet, the slower the disintegration.

[0067] Lubrication of effervescent tablets can contribute to proper and efficient tablet or microparticle production. Non-limiting examples of water-soluble lubricants are glucose, lactose, sorbitol, ascorbic acid, sorbitol, mannitol, sodium benzoate, potassium sorbate, polyethylene glycol or its derivatives, succinic acid, adipic acid, glutaric acid, or clavulanic acid. A tablet press that uses a lubricant spray on the punch can eliminate the need for lubrication.

[0068] Depending on the product, formulators can use pigments (artificial or natural), sweeteners (acesulfame potassium, sodium saccharin, aspartame, and sucralose), and flavors (artificial or natural) to enhance the product or mask the taste or odor derived from the active ingredient.

[0069] The production of effervescent tablets and powders is very similar to that of conventional tablets and powders, but the production must be carried out in areas with very low humidity, which are preferably temperature - controllable. Effervescent particles can be mixed in conventional blending equipment such as ribbon, double - cone, and V - type blenders. All equipment should be well - grounded and kept dry after rinsing for production.

[0070] Wet granulation of the effervescent matrix or fluidized bed drying may be attractive production methods. In the latter, when the effervescent mixture is suspended in a hot drying air stream, water or binder solution is sprayed onto the effervescent mixture.

[0071] Vacuum granulators have also been used to prepare effervescent particles. In this method, water or binder solution is sprayed onto the effervescent mixture during blending.

[0072] Drying is carried out by placing the particles under vacuum and heating through a thermal jacket.

[0073] There are many advantages to using effervescent tablets or particles. One is that CO2 - driven self - blending usually does not require a cumbersome shaking procedure. Another advantage is the flexibility in including flavors, colors, or viscosity enhancers in the material. Yet another advantage is the pre - calculation of the amounts of precursors and prodrugs of active ingredients in the effervescent material.

[0074] The present invention recognizes all these disadvantages associated with the prior - art compositions using chlorine oxides. Specifically, the present invention provides a composition comprising a combination of a solid precursor of an oxidation state of chlorine (OC) and an activator that provides a proton source.

[0075] As described above, in addition to chlorine oxides, the effervescent material also contains two activators. The first activator is a solid carboxylic acid having the formula RN(COOH)m or its salt, where RN is a saturated and unsaturated organic moiety containing 1 - 25 carbon atoms, which is optionally substituted by oxygen and hydrogen to form functional groups that cannot be oxidized by chlorinated species, and m is an integer from 0 - 10, hereinafter referred to as the acid activator.

[0076] The second activator is a base capable of reacting with the acid activator to produce carbon dioxide. The second activator has the general formula

[0077] (M n+ )s((H)1CO3) u ,

[0078] where n, s, t, and u are integers in the ranges n = 1 - 2, s = 1 - 2, and t = 0 - 1, u = 1 - 2, where the sum t + s + n = 1 - 5; M is an alkali metal, alkaline earth metal, or transition metal.

[0079] Preferred basic activators are sodium carbonate or calcium carbonate or sodium bicarbonate.

[0080] Such compositions are useful disinfectants for treating a broad spectrum of microorganisms. Specifically, when the active pharmaceutical ingredient is generated in situ from a stable solid precursor (hereinafter referred to as "API-P" of chlorine oxides), including an acid activator that reacts simultaneously with a buffer solution or gel to a biocompatible pH value, the stability problems in the prior art no longer exist.

[0081] As previously described, the prior art technical solutions have failed to address how to ensure the ionic strength or osmotic pressure of the final antimicrobial solution that is biocompatible with biological fluids. Even further, the prior art has failed to show how to regulate and increase the contact time and persistence of the API in the area of interest for treatment, such as by regulating rheology and fluidity. However, the prior art still fails to provide a relatively simple and effective method to monitor the oxidation state of the API, as well as a visual indication of the location where the API is applied during the mixing of the disinfectant composition.

[0082] Additionally, in some embodiments, the compositions of the present invention may further include the use of a viscosity enhancer (also referred to herein as "VE") and / or include a solid precursor and activator of the oxidation state of chlorine, such as a combination of a diacid or its salt.

[0083] Another embodiment of the present invention is to include a dye in the formulation, preferably a redox-sensitive dye whose color changes with the oxidation state of the chlorine atom, and its advantage solves the disadvantages described for the prior art compositions.

[0084] Specifically, the preferred compositions of the present invention are in solid form. This eliminates any problems associated with the shelf life of hypochlorous acid or chlorine dioxide solutions described in the prior art.

[0085] More specifically, a ready-to-use formulation of chlorine oxide species directly generated from the solid precursor API-P can be released from effervescent powders, tablets or microparticles. Effervescent powders, tablets or microparticles can be used for the preparation, dispensing and long-term stable storage of the prepared compositions consistent with the present invention. Specifically, such effervescent tablets or microparticles described herein can have multiple components required to produce the compositions of the present invention.

[0086] In an example, a solid precursor and activator of the oxidation state of chlorine, such as succinic acid, glutaric acid, adipic acid or pyruvic acid or their salts, a viscosity enhancer and a dye are mixed, and the subsequent composition is generated in the form of a desired disinfectant formulation at the desired time and in situ.

[0087] Further, in addition to its antimicrobial properties, the acids exemplified above are attractive because they cannot be further oxidized by oxidants such as OC, and from a biochemical perspective, because of their endogenous nature at high concentrations.

[0088] Thus, effervescent powders, tablets or microparticles can be effectively used to mix the necessary components to immediately produce an active solution of the API at the point of use. It should be noted that, in order to ensure the ionic strength or osmotic pressure of the final antimicrobial solution to adapt to the osmotic pressure in the area of use in medical applications, the pre-calculated amount of NaCl can be further used to prepare effervescent tablets according to the planned use.

[0089] A preferred embodiment of the present invention is to inhale the solution selected according to the present invention to combat viral infections in the mammalian respiratory system. Therefore, any nebulizer or inhaler that is commonly used to convert a liquid into an aerosol for the treatment of cystic fibrosis, asthma, chronic obstructive pulmonary disease (COPD) and other respiratory diseases or conditions is useful in the present invention. The device generally uses compressed air or ultrasonic energy to generate the atomization of the disinfectant solution. Any type of pressurized metered-dose inhaler (pMDI), dry powder inhaler (DPI), slow mist inhaler (SMI) is particularly useful, for example, as described by Prajapati et al., International Journal of Pharmaceutical Sciences and Research (IJPSR), 2019; Vol. 10(8): 3575-3582. Any electrostatic or non-electrostatic inhaler, such as VORTEX or Pari or Sympotec, can also be used to implement the present invention.

[0090] Effervescent powders, tablets or microparticles have no stability problems, produce a highly broad-spectrum antimicrobial solution when mixing components, and only leave biocompatible inactive chemical species that have been found in human biology or nature.

[0091] As described above, the activation of the API is achieved by using an activator, such as succinic acid or pyruvic acid, which synergistically acts with chlorine dioxide against microorganisms and further maintains the acidity within a controlled pH range between 4 and 8. This novel method and its formulation avoid the lack of long-term stability inherent in chlorine oxide OC in solution (due to its oxidizing ability), because it is not necessary to store the disinfectant composition as an aqueous solution before use.

[0092] Another advantage of the present invention is that other compounds that contribute to the application can be selected. For example, in wound healing applications, it is necessary to increase the viscosity (μ) of the product on the skin to extend the contact time. This is solved by using a water-soluble or soluble viscosity enhancer (VE) that cannot be oxidized chemically by the API, thereby providing improved regulation of the contact time and the persistence of the API in the area of interest for treatment. The VE ensures that the rheology and fluidity are suitable for the corresponding disinfection method and area to produce a solution or gel with complete fluidity. The VE can include, for example, water-soluble gelling agents such as polyacrylic acid, polyethylene glycol or any other oligomer or polymer that cannot be oxidized by the API.

[0093] Additionally, the composition can include one or more dyes identified in a set of redox dyes (also referred to herein as "ROD" or "RODs"), where the color and intensity depend on the oxidation state of chlorine oxide. It should be noted that in addition to providing a visual indication of the oxidation state of the chlorine atom (i.e., through color), the ROD further provides its own antimicrobial effect. This enhances the synergistic effect between the components in the formulation in a novel way. The ROD is capable of maintaining its color for a sufficient period of time to monitor the oxidation activity of the API, chlorine oxide, and further provide a visual indication of the area where the formulation has been applied, thereby addressing the disadvantages of the prior art.

[0094] From the description of the present invention provided below, the advantages of the present invention and additional inventive features will become apparent.

[0095] The chlorine oxide species is represented as OC and has the general formula shown below:

[0096] M r+ [Cl(O) x n n-

[0097] where M can be any alkali metal, alkaline earth metal, or transition metal ion, n is an integer from 1 - 5, x is an integer from 1 - 4, and y is an integer from 1 - 2.

[0098] If M = Na, n = 1, x = 1, then API - P is NaOCl 5. If M = Ca, n = 2, x = 1, then API - P is solid Ca(OCl)2. If M = Na, n = 1, x = 2, then API - P is solid NaClO2. If M = Ca, n = 2, x = 2, then API - P is solid Ca(ClO2)2. In the case of x = 3 or 4, API - P produces more reactive chlorate and perchlorate species.

[0099] A non - limiting example is the instant generation of hypochlorous acid from a mixture of sodium hypochlorite or calcium hypochlorite and sodium succinate in cap 2 according to Figure 1 to provide a ready - to - use solution of API hypochlorous acid with a pH between 4 and 6, optionally with color and viscosity enhancers.

[0100] Another non - limiting example is calcium hypochlorite Ca(OCl)2, which is a stable water - soluble API - P for HOCl, produced and sold on a tonnage scale as a swimming pool disinfectant. This calcium hypochlorite dissolves in water rather slowly, and only calcium hydroxide exists in nature and biology, used as E - number E526 in food, and produces HOCl, which is one of the two active ingredients in the present invention and degrades to Cl - ​and biocompatible species containing hydrogen and oxygen. However, according to the present invention, the effervescent evolution of CO2 significantly increases the dissolution rate of Ca(OCl)2 and other solid components.

[0101] Another preferred embodiment of the present invention is a solid precursor of chlorine dioxide tetrachloride decoxide (TCDO), CAS No. 92047-76-2, a stable solution known as WF10 or OXO-K993, prepared as described by Meuer et al. in CA2616008. This calcium hypochlorite can be prepared by combining a salt of a metal of Group I or II of the chlorite ion ClO2 - with an excess of oxygen in water.

[0102] Thus, one advantage of the present invention is that the solid form precursor API-P in dry and anhydrous quality has no drug stability problems, and thus the present invention solves one of the main technical problems in the prior art.

[0103] The main aspect of the present invention is the combination of API-P with a molecule containing a carboxylic acid functional group -COOH, a sulfonic acid functional group -SO3H, a phosphoric acid functional group -PO3H or a boric acid functional group -B(OH)2, hereinafter defined as an activator of API-P in the formulation. The activator has the general formula R1XO n (R2,) m , where the group R1 can be a group containing 1-10 hydrogenated carbon atoms, optionally substituted by an amino group, an amide group, a carboxyl group or a hydroxyl group. The group X can be a carbon, phosphorus or sulfur atom, n and m can be the integer 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 according to the use and the chlorine species, and can be any compound containing an amino group such as ammonia, an amino acid such as taurine or a therapeutic drug that increases the synergistic potential of the formulation. The activator can be any combination or mixture of two or more compounds defined by the general formula R1XO n R2.

[0104] Preferred non-limiting examples are carboxylic acids R3(COOH)2, where R3 is a straight or branched saturated or unsaturated hydrocarbon chain having 1-24 carbon atoms. Non-limiting examples of the activator 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.

[0105] Taurine is particularly preferred because it is an endogenous amino acid that usually regulates the action of OC in the body and can combine with OC to form an endogenous N-chloro-amino acid, like ClNH-CH2CH2-SO3H, which itself has antibacterial properties.

[0106] Succinic acid, glutaric acid, and pyruvic acid are even more preferred because they are also endogenous substances, have antibacterial properties, have very low toxicity, and form a buffer when mixed with their metal salts and are used as non-limiting examples in the further description of the present invention. In the example, Anuj Purohit and Anand Mohan reported in "LWT - Food Science and Technology" (2019), 116108596, pages 1 - 7, a mixture of pyruvic acid and monosodium succinate, either alone or in the form of a mixture, reduced the bacterial log CFU / g of chicken meat mince by almost log 2.

[0107] An advantage of the present invention is that the solid multi-component product according to the present invention is not affected by stability problems in a pharmaceutical or medical device environment, regardless of temperature, air, humidity, light, oxygen, or other environmental conditions, because API-P is a solid and is commercially available on a large scale.

[0108] Another embodiment of the present invention is that shaking or stirring API-P does not immediately dissolve in water, but is rapidly dissolved or suspended in the liquid phase driven by the effervescent evolution of CO2, and immediately reaches a physiologically acceptable pH and ionic strength in the final solution combined with succinic acid and / or its salts. The products after disinfection are all endogenous species already existing in human biology or nature.

[0109] Another embodiment is that the instant generation of the active disinfectant according to the present invention at the point of use will have a significant and great impact on the versatility of product application. It opens the way for more flexible product packaging technology because the size and amount of the effervescent material in the starting package can be freely varied according to the use.

[0110] Small, stable single-dose disposable, two-component or three-component effervescent material packages will be provided, which are very suitable for tourists, disaster areas, military personnel, or microbial pandemics. Further, the design of large bags, boxes, or cans containing API-P is useful in an agricultural environment, aquaculture, or military operations, suitable for disinfecting larger areas.

[0111] Viscosity enhancers for preparing viscous solutions and gels

[0112] A preferred embodiment of the present invention is to select to include other compounds in the effervescent powder, tablets, or microparticles, which will contribute to the user-friendliness of the product. For wound healing or skin disinfection applications, it is necessary to increase the viscosity (μ) of the product on the skin to extend the contact time. The viscosity enhancer VE solves this medical need.

[0113] Preferred groups of VEs according to the invention are water-soluble gelling agents that do not oxidize the API because all oxygen-containing functional groups in the VE are in their highest oxidation state. The gelling agent provides extended persistence of the API at the site of interest, such as mammalian skin.

[0114] Examples of gelling agents according to the invention include, but are not limited to, polyacrylic acid (carbomer), polyethylene glycol or any other oligomer, polymer or block copolymer thereof. Further, the viscosity enhancer can be selected from poly(acrylic acid)-acrylamidoalkylpropanesulfonic acid copolymer, phosphonopolycarboxylic acid and poly(acrylic acid)-acrylamidoalkylpropane or sulfonic acid-sulfonated styrene terpolymer.

[0115] A preferred VE is polyacrylic acid because each monomer has a carboxylic acid functional group. Since the pKa of PAA is 4.5, using this VE will increase the preferred pH range between 4 - 5.5.

[0116] Accordingly, a preferred aspect of the invention is the use of polymers, such as any type of acrylate copolymer well-known to those skilled in the art, which can function well in the formulations of the invention in the concentration range of 0.01 - 5%. Acrylate copolymers are homopolymers and copolymers crosslinked from acrylic acid and polyvinyl polyethers. Acrylate copolymers have different grafting densities. They differ in their oxidation ability and how many graft chains each polymer has. One possible crosslinking agent is pentaerythritol, which is very stable and thus a good choice for the present invention. Polyacrylic acid (PAA) polymers known to stabilize formulations of H2O2 can be used in the present invention (see Schmucker-Castner and Desai, 1999, "Rheology Modification of Hydrogen Peroxide Based Applications Using A Cross-linked PAA polymer", Int J Cosmet Sci 21(5):313 - 25).

[0117] Polymer-stabilized OC solutions according to the invention have applications in many cases, such as in wound treatment, aseptic packaging, electronics manufacturing, and pulp and paper bleaching. The formulation of the API is compatible with the formulation of a gel or viscous fluid, which can be applied to the target surface, inanimate or representing an infected epithelial mucosa or skin surface of an infected human or animal, to ensure long-term and close contact with the required level of the API. The non-tacky formulation of the API can also be dispersed in the air of an enclosed space in the form of a mist for the purpose of environmental disinfection or for inhalation purposes in the treatment of respiratory diseases.

[0118] In an example, carbomer polyacrylic acid has an increased viscosity at a concentration of 0.01 - 0.1%. If desired, it forms a regular gel in the concentration range of 0.1 - 1%. In order to dissolve VE according to the solution, it must be dissolved in the antimicrobial formulation using a mixer, stirrer or sonicator. The degree of gelling can be further increased using a biocompatible matrix, such as the pharmaceutical buffer triethanolamine or other biocompatible amino alcohols.

[0119] Antimicrobial redox-sensitive antimicrobial dyes as indicators

[0120] In the context of disinfection technology, another innovative and novel aspect of the present invention is the use of redox dyes, hereinafter referred to as ROD for clarity, where the color and intensity depend on the oxidation state of the OC. Even more advantageously, the identified ROD has an antimicrobial effect, increasing the antimicrobial synergy between the components according to the present invention. If the standard half-cell potential of the ROD has a positive value lower than that of the OC, the color of the formulation will be maintained as long as the OC is active. Thus, this color visualizes the area where the formulation has been applied and the area with active OC. This is particularly advantageous when, for example, a formulation according to the present invention is used to treat mastitis, where a large herd of cattle needs to be treated for mastitis; the colored formulation according to the present invention visualizes which animals have been treated. Further, it is also useful to employ the opposite type of indicator, where the color appears when the oxidizing ability of the OC disappears.

[0121] Non-limiting examples of suitable dyes for use in the present invention are non-pH-dependent dyes that are visible in the presence of the OC. Preferred examples are N-phenylanthranilic acid (magenta), N-ethoxychrysoidine (cyan), o-dianisidine (red), sodium diphenylamine sulfonate (red-violet), diphenylbenzidine (violet), diphenylamine (violet) and viologens, which are colorless in the presence of the OC but dark blue in the absence of the OC.

[0122] Another preferred example is the pink dye trisodium (4E)-3-oxo-4-[(4-sulphonato-1-naphthyl)hydrazono]naphthalene-2,7-disulphonate, which is more commonly known as amaranth (E number 123).

[0123] Examples of pH-dependent dyes that are dark blue in the presence of active OC but colorless in the absence of OD are sodium 2,6-dibromophenol-indophenol or sodium 2,6-dichlorophenol-indophenol, sodium o-cresol indophenol, thionine (synthetic Lawes' violet), methylene blue, gentian violet, indigo tetrasulfonic acid, indigo carmine (synthetic indigo disulfonic acid), indigo monosulfonic acid. Examples of dyes that are red or violet in the presence of OC are phenol saffron, saffron T, neutral red, and dialkyl p-phenylenediamine (SPD, violet).

[0124] Many of these dyes have inherent antimicrobial effects, namely methylene blue (MB) and gentian violet (GV), and combinations thereof have been used as antimicrobial dyes in wound dressing foams in combination with polymers like polyvinyl alcohol or polyurethane, for example as described by Edwards in Advances in Wound Care (2016), 5, pp. 11-19.

[0125] One particularly useful class of dyes that can be used in the present invention is the pigmented, redox-active microbial phenazines, nitrogen-containing aromatic compounds of metabolic, ecological and evolutionary significance, see, for example, Chincholkar, S. and Thomashow, L. Microbial Phenazines: Biosynthesis, Agriculture and Health (Chincholkar, S. and Thomashow, L. eds.) 1-243 (Springer, 2014). All of these distinctive features make them attractive microbial metabolites for use as dyes in pharmaceutical formulations. To date, more than 100 natural and more than 6000 synthetic phenazines have been reported to exhibit promising biological activities, including antimicrobial, anticancer, antiparasitic, and insecticidal and biocontrol properties.

[0126] An even more attractive class of phenazines includes bis-N-oxide phenazines, which have stronger antimicrobial properties than their parent phenazines. Most of these compounds are natural compounds produced by bacteria and are heteroaromatic N-oxide compounds, hereinafter denoted as HANOX. In addition to being redox dyes, RODS, HANOX compounds are also useful in the present invention because their color depends on the oxidation state of OC. At the same time, HANOX in the prior art has been shown to have broad-spectrum antimicrobial activity, for example, as described by Leimgruber et al. in US3822265.

[0127] Furthermore, the phenazine derivatives provided by US3822265 have broad-spectrum fungicidal activity. Specifically, it exhibits high activity against a 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, the phenazine derivatives can be particularly used to treat animal diseases caused by microorganisms in agriculture.

[0128] The surprising discovery of these derivatives is that they have no harmful effect on body tissues under the use conditions, which makes them particularly suitable for topical application, preferably used in an amount ranging from 0.05% to 1.0% by weight of the composition.

[0129] It has particular value in topical applications, for example, in solid or gel formulations, including divided powders and particulate materials, and in liquid formulations, including solutions, suspensions, concentrates, tinctures, slurries, and aerosols, creams, gels, gelatins, ointments, and pastes.

[0130] Viktorsson et al. described a group of even more preferred HANOX in WO 2015063516A2 and WO 2018109504A1 that can be used in the present invention. This technology describes RODs with redox properties and broad-spectrum antimicrobial properties. As described by Viktorsson et al. in Bioorg. Med. Chem. (2017) 25, pp. 2285-2293, this specific class of HANOX is particularly attractive because, compared to the compounds described in US3822265, it shows lower toxicity to mammals and a lower risk of toxic reactions when used in infected hosts.

[0131] Methylene blue is another particularly preferred dye that can be used in the present invention because the FDA has approved it as an excipient in pharmaceutical formulations, and it has antibacterial properties, and its effect as a therapeutic agent can be enhanced using photodynamic therapy.

[0132] Effervescent tablets that can be used in the present invention

[0133] Figure 1 is a schematic, non-limiting, and exemplary diagram of an effervescent tablet showing a disinfectant solution for instant generation of an API with an excipient in an aqueous solution according to the embodiments and methods of the present invention.

[0134] Effervescent tablets are used in different medical, household, and agricultural applications. In many of these applications, the antimicrobial solution generated from the effervescent material is useful in the present invention and can be generated in a two-chamber or multi-chamber bottle, bag, syringe, inhaler, hand disinfection device, spray bottle or flask or can, made of hard or soft materials such as 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 household environment without the need for complex mixing procedures and can be stored at ambient temperature. It can be combined with an automatic dispensing system, is easy to label, and is automatically mapped by barcode.

[0135] The effervescent tablets according to the present invention are composed of pre-calculated amounts of components and can be designed to eliminate mixing errors to avoid unwanted exposure of patients and personnel and to comply with The Joint Commission and USO 797 guidelines.

[0136] Use of the present invention for antimicrobial purposes in photodynamic therapy

[0137] The elimination of bacteria using antimicrobial photodynamic therapy (aPDT) has shown the use of alternative treatment modalities in the treatment of peri-implantitis. In "Photodiagnosis and Photodynamic Therapy" (2019), 25, pp. 7-16, Huang et al. described the dose-dependent and pH-dependent bactericidal effects of methylene blue (MB)-mediated aPDT on Gram-negative (Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans) and Gram-positive (Streptococcus mutans) bacteria on sandblasted, large-grit and acid-etched (SLA)-pretreated titanium alloys. However, the test formulation did not contain OC to even further enhance the therapeutic effect.

[0138] Therefore, another preferred embodiment of the innovative formulation of the present invention comprising OC, succinic acid or its salt, and optionally a viscosity enhancer is to include a ROD exemplified as methylene blue for photodynamic therapy, such as for improving wound healing or bacterial infection in mammals. In this case, the application site of the product according to the present invention can be irradiated with light having a wavelength suitable for generating the photodynamic effect of the dye.

[0139] In "Photodiagnosis and Photodynamic Therapy" (2018), 23, pp. 347-352, Souza et al. used photodynamic therapy to show the antimicrobial activity of hypochlorite solution and the antimicrobial activity of a reciprocating instrument associated with photodynamic therapy against the root canals infected with Enterococcus faecalis. However, the test solution did not have an antibacterial dye. These techniques are incorporated herein by reference.

[0140] Use of the present invention in agriculture

[0141] In agriculture, specifically in animal farms, multiple infectious diseases caused by bacteria, viruses, and fungi affect the daily operation of the farm and the cost of running the facility. In these cases, the formulations designed according to the present invention have a therapeutic or prophylactic effect and are particularly suitable for skin infections.

[0142] An important example is mastitis in cows, which costs the US dairy industry approximately $1.7 - $2 billion annually. Effective and environmentally friendly treatment of mastitis has proven difficult because milk from dairy cows cannot be sold until the residual drugs have left the system after a long course of antibiotic treatment. No vaccine has been effective because infections in the cow's udder and teats are far from the animal's main bloodstream. To mark cows that have been treated, dairy workers use tape strips to alert and mark the treated cows.

[0143] Therefore, a preferred aspect of the present invention is the use of a gel or viscous solution for treating mastitis, the gel or viscous solution comprising OC, succinic acid or its salts, or other preferred acid activators or their salts, such as the viscosity enhancer VE listed above and the ROD exemplified as methylene blue. The colored gel stays in the area of the udder and teats, succinic acid can penetrate into the skin of the teat, and this color makes it unnecessary to use tape strips. Additionally, the applied gel can be irradiated with light of a suitable wavelength to increase the therapeutic effect of the gel. In this case, steps 1 - 4 and step 6 are carried out to produce a ready - to - use formulation.

[0144] Use of the present invention in aquaculture

[0145] Water quality is a prerequisite for successful aquaculture of aquatic animals, exemplified by fish, oysters, shrimps, and prawns. Open water systems typically introduce organisms such as viruses, bacteria, lice, protozoa, fungal pathogens, algae, and parasites. Common viral infections that cause high mortality rates in attractive aquatic food species are Koi HerpesVirus Disease, Pancreatic Disease (PD), and Infectious Salmon Anemia (ISA). Appropriate water quality or a sufficient quantity of pure water is usually not available. Culturing devices in the prior art generally have no way to prevent these infectious species from approaching and affecting the cultured species. Further, once infected, there is no highly effective cure to provide an efficient therapy against these diseases.

[0146] A preferred embodiment of the chlorine oxide species OC according to the present invention is an effective treatment for all these infections and harmful organisms and cells. The immediate formulation of OC is very effective in controlling these waterborne pathogens. In an example, chlorine dioxide is a broad-spectrum biocide that effectively solves the problems defined in the prior art. The formulation of the present invention is even used in special tanks to repeatedly treat, for example, farmed salmon without harming the gills or any other part of the farmed species, while having a destructive effect on the disease-causing microorganisms. In these applications, a preparation sequence is used in which the API-P, which is NaOClO2 or Ca(OClO2)2 in steps 1-3, is mixed with a pre-calculated amount of succinic acid into an effervescent tablet.

[0147] Anti-war application of the present invention

[0148] Another preferred embodiment of the chlorine oxide species OC according to the present invention is to reduce or eliminate warfare agents, such as nerve gases, such as the G series, exemplified by GA, GB (sarin), GD (soman), GF (cyclosarin), GV, the V series, exemplified by VE, VG, VM, VR, and VX, Novichok, carbamates, pesticides, protein toxins such as ricin, or biological warfare bacteria, viruses, or fungi.

[0149] All these species have the common feature that they can be oxidized by the chlorine oxide species according to the present invention. For example, the nerve gases GD or soman have a phosphoryl halide group that can be oxidized by the oxidation chlorine species into harmless species, as described by Xu et al. in E3S Web of Conferences 267, 02043 (2021), pages 1-6.

[0150] However, in this example, an unstable hypochlorous acid solution was used. According to the present invention, the stability problem has been solved by using a solid effervescent material as an immediate precursor of the reaction solution.

[0151] An example of the anti-war use of the solution produced in the present invention is to generate a fog or mist of oxidized aerosol droplets that can stay indoors or outdoors for a long time to protect the living organisms in an area or location from the effects of warfare agents.

[0152] Antiviral use of the present invention

[0153] The methods disclosed herein also allow for improving methods of exposing contaminated surfaces, devices such as medical devices, any furniture surface, door handles, fixtures, clothing, or persons to a disinfectant formulation by atomizing or vaporizing an API into a confined space. This is possible because the degradation state and quality of the disinfectant formulation are known since it is not stored as a solution but is freshly prepared from solid precursors at the location of interest.

[0154] This procedure ensures the dispersion of the active agent into crevices and microenvironments and even onto persons suspected of being contaminated with infectious tissue or body fluids. The evaporation of these formulations can have beneficial therapeutic or prophylactic effects against viral, bacterial, or fungal infections.

[0155] Kim et al. studied the effects of low-concentration hypochlorous acid in nasal irrigation solutions against bacteria, fungi, and viruses in Laryngoscope (2008), 118, pp. 1862-1867. The formulation was also used to inactivate human rhinovirus (HRV) in nasal epithelial cells in vitro with significant virucidal effects. Similar formulations were used in in vitro experiments and had good effects against avian influenza virus as described by Hakim et al. in J. Vet. Med. Sci. (2015), 77, pp. 211-215.

[0156] Hypochlorous acid has also been used clinically in the upper respiratory mucosa of patients with rhinitis. Cho et al. reported improved results of low-concentration hypochlorous acid after nasal irrigation in children with chronic rhinosinusitis 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 of mammals.

[0157] In Robert Northey's US10,342,825B, a low-pH antimicrobial solution contains 5 mg / L to 200 mg / L hypochlorous acid and water, where the pH of the solution is 5.6 and is stabilized by a phosphate buffer. The solution can be evaporated using a nebulizer to be distributed on surfaces and tissues. However, the disadvantages of the formulations used in the prior art are the same as those described above.

[0158] In Terry's WO 2019 / 222768, a method for inactivating an infectious agent, which is a resistant virus, an oncogenic virus, a chemically resistant non-enveloped virus, or an infectious agent present in mucosal or epithelial surfaces. The formulation contains the infectious agent and a non-buffered electrolytic hypohalous acid composition. However, also in this case, the weaknesses of the formulations used herein are the same as those defined above.

[0159] The present invention has solved many problems in the prior art. OC is now combined for the first time with an activator from a solid precursor and NaCl to maintain biological osmotic pressure for antiviral use of the resulting formulation.

[0160] Exposure can be affected without regard to toxicity or corrosiveness, which accompany existing inactivation methods for highly infectious and resistant infective agents. A preferred embodiment is to eradicate, minimize or prevent the progression of viral infection in the upper respiratory tract, such that the immune system has time to produce antibodies in response to the virus.

[0161] Accordingly, the systems and methods of the present invention provide chlorine oxide OC as a means of treating respiratory viral infections. The compositions of the present invention are capable of treating SARS, MERS and other infections, including but not limited to SARS CoV-2 infection. This is now facilitated for the first time by combining an immediate precursor of the API with an effervescent tablet according to the present invention, since there is no need to evaluate the lack of activity of the solution stored under environmental conditions.

[0162] Specifically, an inhalable hypochlorous acid formulation of OC; an activator such as succinic acid or pyruvic acid; an excipient that modulates the rheology of the final solution; an osmotic pressure regulator such as sodium chloride - such immediate formulations can now be prepared on-site and can use methods of delivery via nebulizers such as soft mist inhalers, jet nebulizers, ultrasonic nebulizers and vibrating mesh nebulizers. In use, the inhaler and nebulizer atomize the composition of the present invention for delivery by inhalation.

[0163] Formulations that can be used to generate an aerosol can be provided in the form of a dry powder, a solution or a suspension. Fine droplets, sprays and aerosols can be delivered via intranasal or intralung pump dispensers or squeeze bottles. The composition can also be inhaled via an inhaler such as a metered dose inhaler or a dry powder inhaler. The composition can also be inhaled via a nebulizer such as an ultrasonic nebulizer, delivering a composition of OC and succinic acid directly to the respiratory tract via an inhalable formulation. This can prevent and treat respiratory infections caused by viruses and other microorganisms. According to the present invention, the formulations as described herein are safe and effective for the prevention and treatment of viral infections.

[0164] The compositions of the present invention can also include 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 may be specifically suitable for different formulations of the composition, such as whether used as droplets or drops, suspensions or another form for pulmonary delivery.

[0165] The formulation for atomization can be provided in the form of a dry powder, a solution or a suspension. The composition can be delivered by various devices known in the art for administering drops, droplets and sprays. The composition can be delivered by a dropper, a pipette or a dispenser. Fine droplets, sprays and aerosols can be delivered by an intranasal or pulmonary pump dispenser or a squeeze bottle.

[0166] Intranasal delivery can be provided by a nasal spray device. Thus, the formulation according to the invention can be designed as a nasal spray. The nasal spray is sprayed into the nose and delivered to the respiratory tract.

[0167] The soft mist inhaler pressurizes a liquid container using the mechanical energy stored in a spring by user actuation, so that the contained liquid is ejected from the nozzle in the form of a soft mist for inhalation. The soft mist inhaler does not rely on a gas propellant or electricity for operation. The average droplet size in the soft mist inhaler is about 5.8 microns.

[0168] The jet nebulizer is the most commonly used and can be referred to as a nebuliser. The jet nebulizer uses a compressed gas (e.g., air or oxygen) to atomize a liquid medicament as it is released at high speed through it. The atomized droplets of the resulting therapeutic solution or suspension are then inhaled by the user for treatment. The compressed gas can be pre-compressed in a storage container or can be compressed on demand by a compressor in the nebulizer.

[0169] The ultrasonic nebulizer relies on an electronic oscillator to generate high-frequency ultrasonic waves, which atomize the drug for inhalation when directed through a reservoir of a therapeutic suspension of the solution.

[0170] The vibrating mesh nebulizer uses the vibration of a membrane with thousands of holes on top of a liquid reservoir to atomize a fine droplet mist for inhalation. The vibrating mesh nebulizer avoids some of the disadvantages of ultrasonic nebulizers, thus providing more efficient aerosol generation, reducing the processing time and reducing the heating of the atomized liquid.

[0171] A synergistic composition of glutaric acid and hypochlorous acid is used to treat viral infections. The glutaric acid component is particularly effective in penetrating into tissues, while hypochlorous acid is particularly effective in treating infections on the outer surface of tissues. As described above, these compositions are effective in treating and preventing respiratory tract infections.

[0172] The disclosed compositions are particularly effective because balancing the concentrations of hypochlorous acid and glutaric acid with NaCl allows for the safe treatment of viruses. The exact balance depends on the formulation, the treatment site, and even the desired surface penetration. Hypochlorous acid can be present at about 5 ppm up to about 1000 ppm or more. Different uses, different 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, and preferably about 1.0%. By balancing these two components, the composition can have a dual effect of treating both on and under the surface of the tissue to which it is applied.

[0173] In the case where OC is hypochlorous acid HOCl, an immediate composition of OC at a concentration of about 15 - 200 ppm is generally sufficient to treat an infected lung. In the case where OC is chlorine dioxide OCl2, a concentration of 0,1 - 5 ppm is generally sufficient.

[0174] In some cases, in order to completely destroy the virus or prevent the virus from entering the respiratory tract, the composition should be in contact with the virus for a longer period of time, from a few seconds to a few minutes, to an hour or more. Thus, in certain embodiments, the composition is in the form of a gel that allows for longer contact with the site of infection.

[0175] The use of the composition in combination with known antiviral therapies can increase the efficacy of the composition. In some embodiments, the methods of the invention further comprise administering (simultaneously or sequentially with the composition of the invention) one or more doses of an antiviral agent. 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 inhibitors, neuraminidase inhibitors, interferons, ribavirin, nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors, non-structural protein 5a (NS5a) inhibitors, chemokine receptor antagonists, integrase strand transfer inhibitors, protease inhibitors, and purine nucleosides.

[0176] The composition can also be used in combination with known antimicrobial therapies.

[0177] In some embodiments, the method of the present invention further comprises administering, simultaneously or sequentially with the composition of the present invention, one or more doses of an antibiotic substance, including but not limited to ciprofloxacin, β-lactam antibiotics such as ampicillin or carbapenems, azithromycin, cephalosporin, doxycycline, fusidic acid, gentamycin, linezolid, levofloxacin, norfloxacin, ofloxacin, rifampin, tetracycline, tobramycin, vancomycin, amikacin, ceftazidime, cefepime, trimethoprim / sulfamethoxazole, piperacillin / tazobactam, aztreonam, meropenem, colistin or chloramphenicol.

[0178] In some embodiments, the method of the present invention further comprises administering one or more doses of an antibiotic substance selected from the group of antibiotics including, but not limited to, aminoglycosides, carbacephem, carbapenems, first-generation cephalosporins, second-generation cephalosporins, third-generation cephalosporins, fourth-generation cephalosporins, glycopeptides, macrolides, monobactam, penicillins, polypeptides, quinolones, sulfonamides, tetracyclines, lincosamides, and oxazolidinones. In some embodiments, the method of the present invention comprises administering a non-antibiotic antimicrobial substance including, but not limited to, sertraline, thioridazine in racemic and stereoisomeric forms, benzoyl peroxide, taurolidine, and hexetidine.

[0179] The dosing regimen of the composition can include the amount, frequency, and duration of exposure to the composition. The dosing regimen can depend on the severity of the infection or on the regimen prescribed for the treatment or prevention of a viral infection.

[0180] The composition can be administered as a single daily dose or multiple doses, such as 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 may depend on the frequency, amount, or even severity of the infection.

[0181] Depending on the nature of the OC, the total daily amount of the API formed from the solid precursor in the immediate solution can range from 0.01 - 1000 mg. The actual dose can vary depending on the specific composition administered, the mode of administration, and other factors known in the art.

[0182] The composition can be administered to any part of the respiratory tract, such as the respiratory epithelium, nasal cavity, nasal epithelium, pharynx, esophagus, larynx, epiglottis, trachea, carina, bronchi, bronchioles, or lungs. Administering the composition to the respiratory tract treats and prevents any disease or disorder transmitted by a virus.

[0183] In certain other embodiments, for example, the compositions of the present invention can be used to disinfect entire rooms, facilities, medical devices, and surgical instruments. The supply of medical devices is typically initially sterile, but may require additional or subsequent cleaning and disinfection or sterilization. Specifically, it is particularly important to sterilize or disinfect reusable medical devices before reuse using any known technique. The composition can be applied to the use of medical devices. For example, the composition can be applied by wiping or spreading it onto the surface of the device, spraying an aerosol or mist form of the composition onto the device, immersing the device in a container containing a certain volume of the composition, or placing the device, such as from a faucet, into the flow of the composition. Additionally or alternatively, medical devices and surgical instruments can also be stored immersed in the composition and taken out for use.

[0184] In summary, as known in the prior art, it has been found that the disinfection efficacy of the fully instant formulations according to the present invention is greater than that of its individual components. The performance difference is easily observable over a wide concentration range.

[0185] Additionally, since both chlorine dioxide chemical species and glutaric acid are toxic at high concentrations, the prior art has taught not to use these agents on the skin or other tissues, except in trace amounts. Thus, the present invention is surprisingly safe and effective when used in a controlled manner as described above.

[0186] Some of the disclosed compositions contain 2% or more glutaric acid and have been shown to be safe and effective for treating the skin and other tissues when combined with OC. It has been found that the OC in these compositions has a modulating effect on glutaric acid.

[0187] This allows the composition to utilize the disinfection properties of glutaric acid without causing harm to the tissue.

[0188] General procedure for preparing a dry solid mixture of pre-calculated amounts of API-P and NaCl for use in effervescent tablets.

[0189] Example 1. Preparation of OC powder and effervescent powder formulations

[0190] a) Preparation of powdered Ca(OCl)2 as part of a powder / effervescent powder / effervescent tablet / particle composition

[0191] I. 100 g of calcium hypochlorite particles were continuously introduced into a ceramic mortar (Porsgrunn Porselen, Norway). The particles were ground in several rounds using the mortar and pestle. The powder was sieved through masks of ≤75 μm, ≤150 μm, ≤250 μm, ≤355 μm, and ≤500 μm (Retsch, Haan, Germany). The powder material was stored in various containers made of the following materials: PET, amber glass, PE.

[0192] II. 100 g of calcium hypochlorite particles for dry grinding were continuously introduced into 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.

[0193] III. 100 g of calcium hypochlorite particles for dry grinding were continuously introduced into 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.

[0194] b) Preparation of powdered sodium hydroxide (NaOH) as part of a powder / effervescent powder / effervescent tablet / particle composition

[0195] I. 100 g of sodium hydroxide agglomerates were continuously introduced into a ceramic mortar (Porsgrunn Porselen, Norway). The particles were ground in several rounds using the mortar and pestle. The powder was sieved through masks of ≤150 μm, ≤250 μm, ≤355 μm, and ≤500 μm (Retsch, Haan, Germany). The powder material was stored in various containers made of the following materials: PET, amber glass, PE.

[0196] II. 100 g of sodium hydroxide agglomerates for dry grinding were continuously introduced into 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.

[0197] III. 100 g of sodium hydroxide agglomerates for dry grinding were continuously introduced into 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.

[0198] c) Preparation of a powder formulation using glutaric acid as an acid activator

[0199] Mix 1.65 g of glutaric acid 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) by volume to obtain a pre-calculated powder formulation that is easily mixable with water to release a stable hypochlorous acid solution.

[0200] d) Prepare a powder formulation using glutaric acid as an acid activator

[0201] Mix 1.48 g of succinic acid 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) by volume to obtain a pre-calculated powder formulation that is easily mixable with water to release a stable hypochlorous acid solution.

[0202] e) Prepare an effervescent powder formulation using glutaric acid as an acid activator

[0203] Mix 3.30 g of glutaric acid with 1.60 g of sodium bicarbonate and 220.9 mg of powdered Ca(OCl)₂ (particle size ≤ 150 μm, ≤ 250 μm, ≤ 355 μm, ≤ 500 μm) by volume to obtain a pre-calculated effervescent powder formulation that is easily mixable with water to release a stable hypochlorous acid solution. Measure the effervescence time by dissolving the effervescent powder in a beaker containing 100 ml of distilled water at 20 °C. Measure the effervescence time with a timer; the effervescence ends when the solution becomes clear and has no particles.

[0204] f) Prepare an effervescent powder formulation using succinic acid as an acid activator

[0205] Mix 2.95 g of succinic acid with 1.61 g of sodium bicarbonate and 221.9 mg of powdered Ca(OCl)₂ (particle size ≤ 150 μm, ≤ 250 μm, ≤ 355 μm, ≤ 500 μm) by volume to obtain a pre-calculated effervescent powder formulation that is easily mixable with water to release a stable hypochlorous acid solution. Measure the effervescence time by dissolving the effervescent powder in a beaker containing 100 ml of distilled water at 20 °C. Measure the effervescence time with a timer; the effervescence ends when the solution becomes clear and has no particles.

[0206] Example 2. Powder flow measurement

[0207] Add 100 g of powdered calcium hypochlorite with particle sizes ≤ 355 μm and ≤ 500 μm to a dry funnel ( (Outlet diameter = 15 mm), the bottom opening of the funnel was blocked with a plastic cap. The bottom opening of the funnel was opened to measure the time required for all the samples to flow out. The flowabilities of particle sizes ≤355 μm and ≤500 μm were 35.7 g / s and 37.4 g / s, respectively. The angles of repose of particle sizes ≤355 μm and ≤500 μm were calculated to be 29.5° and 26.8°, respectively.

[0208] Example 3. Dissolution time of powdered calcium hypochlorite with particle sizes ≤250 μm, ≤355 μm, and ≤500 μm

[0209] I. 58 ± 4 mg of powdered materials of various sizes were separately dissolved in 100 mL of 0.25% wt. acetic acid / acetic acid buffer. At 2500 rpm, particle sizes ≤250 μm dissolved within 10 seconds, ≤355 μm within 15 seconds, and ≤500 μm within 1 minute and 25 seconds.

[0210] II. 58 ± 4 mg of powdered materials of various sizes were separately dissolved in 100 mL of 3.0% wt. acetic acid / acetic acid buffer. At 2500 rpm, particle sizes ≤250 μm dissolved within 10 seconds, ≤355 μm within 25 seconds, and ≤500 μm within 25 seconds.

[0211] III. 570 ± 40 mg of powdered materials of various sizes were separately dissolved in 100 mL of 0.25% wt. acetic acid / acetic acid buffer. At 2500 rpm, particle sizes ≤250 μm dissolved within 15 seconds, ≤355 μm within 45 seconds, and ≤500 μm within 1 minute and 35 seconds.

[0212] IV. 570 ± 40 mg of powdered materials of various sizes were separately dissolved in 100 mL of 3.0% wt. acetic acid / acetic acid buffer. Repeated with 3.0% acetic acid / acetic acid buffer. At 2500 rpm, particle sizes ≤250 μm dissolved within 15 seconds, ≤355 μm within 15 seconds, and ≤500 μm within 20 seconds.

[0213] Example 4. Preparation of effervescent tablets

[0214] a) Preparation of particulate materials

[0215] Useful preferred carboxylic acids and bases according to the invention have the following molecular weights: sodium hypochlorite (mw: 74.44 g / mol), calcium hypochlorite (mw: 142.98 g / mol), sodium chlorite (mw: 90.44 g / mol), calcium chloride (mw: 157.89 g / mol), succinic acid (mw: 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).

[0216] The process temperature of 55 °C is determined by the physicochemical constraints of the components; in the example, dimethyl polyethylene glycol with an average molecular weight of 6000 D (PEG6000 - di - Me, di - methyl ether) and sodium bicarbonate. The effervescent powder contains 100 g of a stoichiometric effervescent system formed by anhydrous succinic acid (43.2%) / sodium bicarbonate (56.8%), as a medium - coarse powder or a very fine powder, and the fine - powdered PEG6000 - di - Me is added to the 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), first adjusted to 55 °C, 61 °C and 123 m3 / h. The particles are fused by melting the PEG6000 - di - Me for 5 minutes, 15 minutes or 30 minutes to obtain a particulate material. After cooling for 30 minutes at ambient temperature and 30% - 63% relative humidity (RH), the particulate material is screened using an oscillating granulator (Erweka FGS, Frankfurt, Germany) fixed at medium speed (II) and equipped with an 800 - μm sieve.

[0217] Measurement of the physical properties of the particulate effervescent material

[0218] Measurement of particulate flowability and density The particulate flowability and density are measured using a precisely weighed 100 - g sample. The particulate flowability is measured using a flowmeter consisting of a standard funnel and a timer. The tapped density is determined using a volumeter (Jel / Stav 2003A, Ludwigshafen, Germany).

[0219] The particle size distribution was analyzed using 100 g of the particles and vibrating a Siever Retsch (Haan, Germany) fitted with a series of sieves from the European Pharmacopoeia (3rd edition) (710 μm, 500 μm, 355 μm, 250 μm, 180 μm, 125 μm) (amplitude 1.5; 10 minutes). The mean particle size was determined graphically using a log-normal plot.

[0220] The effervescence time was determined at 20 °C using 3 g of the particles accurately weighed into a beaker containing 200 ml of distilled water. The effervescence time was measured using a timer; effervescence ended when the solution became clear and free of any particles.

[0221] Tablet pressing studied the particle properties that formed tablets with a crushing resistance between 70 N and 120 N when pressed in a single-punch press. After mixing for 10 minutes at 28 rpm in a Turbula mixer, 700 g of the effervescent particles were mixed with sodium benzoate or silicified sodium benzoate to obtain a particle mixture that was easy to press into tablets. Silicified sodium benzoate was manufactured by mixing 600 g of very fine sodium benzoate powder with 40 g of Silbione (silicone) at low speed in a Kenwood planetary mixer for 30 minutes (II). The mixture was screened through a 125-μm sieve. Silbione is an oil lubricant that can enhance the efficiency of sodium benzoate. Tablets were manufactured in a workshop with air conditioning adjusted to 30%–63% RH and 22 °C–61 °C using a single-punch press (Frogerais OA, Evry Lisses, France) equipped with a chrome-plated punch with a diameter of 24 mm. The tablet weight, crushing resistance (Erweka TBH 28, Heusenstamm, Germany), friability (Pharmatest PTF 1E, Haiberg, Germany), and effervescence time were evaluated using the European Pharmacopoeia method (10th edition). The water content, carbon dioxide content, pH of the solution, tablets, and solutions were evaluated using the same method as for the particles. Visual evaluations were also made of the adhesion to the punch surface and die wall, capping, and sliding friction on the die wall. All formulations were pressed under constant conditions (depth of the lower punch in the die 6.98 mm and distance the upper punch penetrated the die 6.04 mm) to select tablets that weighed more than 2 g and had a crushing resistance of 70 N to 120 N and no processing problems during pressing. Physicochemical

[0222] The pH was determined using an Aquadata APH 1000 pH meter. The carbon dioxide content was determined using a sensitive balance Mettler PG 503S (Viroflay, France) for the carbon dioxide content of either 3 g of the microparticles weighed precisely or one tablet in 100 ml of diluted sulfuric acid (R) (18, 19). The results were expressed as the weight loss of the sample at the end of effervescence (mg CO2 / g ES).

[0223] Using this procedure, the tablets will generally have a water content of <0.1, an average particle size of 510 μm, a CO2 content of the microparticles of 300 mg CO2 / g, an effervescence time of 70 seconds per minute, a pH of the resulting solution of 5.6, and a typical dissolution volume of 5 mL for each tablet in water or an environmentally pharmaceutically acceptable carrier.

[0224] Example 5 In vitro anti-biofilm effect of the example Three different test solutions of HOCl and succinic acid.

[0225] The three different test solutions were produced from effervescent tablets. All three test solutions were produced from effervescent tablets.

[0226] Experimental setup

[0227] Test organisms: Pseudomonas aeruginosa or Staphylococcus aureus wild-type strains Biofilm type: 48-hour or 24-hour biofilms grown on semi-permeable membranes placed on solidified media supplemented with 0.5% glucose. In the case of the 48-hour biofilm, the membrane with the biofilm was transferred to a fresh plate after 24 hours.

[0228] Initial viable cell count: 5 x 10 9 colony-forming units (CFU)

[0229] Treatment method: The membrane with 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 the antimicrobial solution was pipetted onto the gauze layer. This treatment was carried out at room temperature for 2 to 3 hours or 4 to 6 hours. In the case of the 4 to 6-hour treatment, the gauze layer was replaced with a fresh gauze layer containing 1 ml of the sample solution 2 or 3 hours after the start of the treatment.

[0230] Evaluation method: The gauze layer was discarded, and each membrane with the 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 μl of each dilution was spotted onto LB plates for viable CFU counting.

[0231] Results and conclusions

[0232] Figure 2 shows the results obtained using the sample solution. In a 200 ppm HOCl solution, increasing the concentration of the acid activator 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. Comparing these three test solutions with 4 different commercially available competing wound healing products, all products showed only a slight effect on Staphylococcus aureus biofilms. An even stronger effect was shown on biofilms from Pseudomonas aeruginosa. It was concluded that hypochlorous acid and succinic acid act synergistically and effectively at concentrations of pH 4 - 7, which have been shown to be safe in other studies.

[0233] Example 6 In Vivo Toxicity Study

[0234] Example 6.1: 7 - day Inhalation Toxicity Study in Rats.

[0235] A 7 - day inhalation toxicity study was conducted on rats as described by Kogel et al. in Food ChemToxicol. 2014 Jun;68:204 - 17. The rat inhalation study was conducted according to the Organization for Economic Cooperation and Development (OECD). The test solution was generated from effervescent tablets. In accordance with Test Guideline 412, Sprague - Dawley rats were exposed to filtered fresh air (sham) as a reference or the test solution. The care and use of animals complied with the American Association for Laboratory Animal Science Policy (1996). All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC). Histopathological evaluations were performed at specified anatomical sites in the nose and left lung according to a defined grading system. Free lung cells in bronchoalveolar lavage fluid were determined by flow cytometry, and inflammatory mediators were measured by multiplex analyte profiling (MAP). For the systems toxicology approach, RNA samples were obtained from specific sites in the respiratory tract, namely the respiratory nasal epithelium (RNE) and the lung. For lung RNA isolation, the respiratory epithelium of the main bronchi and lung parenchyma was isolated by laser capture microdissection (LCM), further processed, and analyzed on a whole - genome Affymetrix microarray ( Rat Genome 230 2.0 Array). No significant perturbations related to inflammation, cellular stress, or cell proliferation in the bronchi or lung parenchyma were found.

[0236] Example 7. Treatment of mastitis

[0237] For the application of the color indicator in step 4 to add information in the treatment procedure, such as in the indication of the oxidative activity of the API or for the application of indicating the oxidative activity of the API, the compartment containing ROD is included in the procedure.

[0238] Example 8. Clinical antiviral therapy

[0239] The medicine cup of the Gima Aerosol Corsia nebulizer is filled with 5 mL of the test solution generated by the effervescent tablets. The mouth of a patient with a coronavirus lung infection is connected to the hose, and the mask is connected to the activated nebulizer. After breathing for 10 - 15 minutes, the fluid is used up and the nebulizer is turned off. The patient is monitored for several hours to ensure that the treatment does not produce side effects. The potential side effects on the patient's mucosa and cilia are studied.

[0240] Incorporated by reference

[0241] Any and all references and citations to other documents such as patents, patent applications, patent publications, journals, books, papers, web content, etc. throughout this disclosure are hereby incorporated by reference in their entirety for all purposes.

[0242] Equivalents

[0243] The present invention may be embodied in other specific forms without departing from its spirit or essential characteristics. Accordingly, the foregoing embodiments are to be considered in all respects illustrative rather than limiting the invention described herein.

Claims

1. An antimicrobial composition comprising: a solid chlorine oxide species salt according to the following 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; an acid activator having the following formula: RN(COOH) m or a salt thereof, wherein RN is a saturated or unsaturated organic moiety containing 1 - 25 carbon atoms, the organic moiety optionally being substituted by oxygen or hydrogen to form a functional group that cannot be oxidized by the chlorine species, and m is an integer from 0 to 10; a pharmaceutically acceptable diluent, adjuvant or carrier; and a base activator capable of reacting with the acid activator to produce carbon dioxide, the base activator having the following formula: (M′ z+ ) s ((H) t CO3) u wherein 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.

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

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

4. The composition according to claims 1 to 3, wherein the acid activator is selected from the group consisting of: monobasic acids, dibasic acids, tribasic acids, propionic acid, lactic acid, succinic acid, glutaric acid, pyruvic acid, citric acid, malic acid, oxaloacetic acid, tartaric acid, adipic acid, fumaric acid, pimelic acid, suberic acid, diacids and their derivatives.

5. The composition according to claims 1 to 4, wherein the base activator is selected from the group consisting of: sodium bicarbonate, potassium bicarbonate, sodium carbonate and potassium carbonate.

6. The composition according to claims 1 to 5, further comprising a water - soluble polyol binder.

7. The composition according to claims 1 to 6, further comprising a water - soluble polyol binder selected from the group consisting of: monosaccharides, disaccharides, polysaccharides or monohydroxy acids or polyhydroxy acids.

8. The composition according to claims 1 to 7, further comprising a lubricant.

9. The composition according to claims 1 to 8, further comprising a lubricant selected from the group consisting of: glucose, 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 any one of claims 1 to 9, wherein the osmotic pressure of the composition is in the range of about 0.1 mOsm to about 500 mOsm.

11. The composition according to any one of claims 1 to 9, wherein the pH of the composition is between 4 and 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)-acrylamidoalkyl propane sulfonic acid copolymer, phosphonated polycarboxylic acid, poly(acrylic acid)-acrylamidoalkyl propane 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 the chlorine oxide compound.

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

17. The composition according to claims 15 to 16, 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 components are contained within an effervescent powder, microparticles or tablets.

19. A method for treating pathogens, the method comprising administering the composition according to claim 1.

20. The method according to claim 19, wherein the composition is administered orally, by inhalation, topically or intravenously.

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