Broad spectrum antimicrobial formulations comprising lysozyme and methods of use thereof

Abstract

To provide broad spectrum microbicide formulations that can be used to treat or prevent a wide range of diseases of bacterial etiology, and methods of treating or preventing bacterial infections in mammals.SOLUTION: A pharmaceutical formulation for administration to a mammal comprising: from about 2% to about 80% by weight lysozyme, from about 2% to about 40% by weight a pharmaceutically acceptable chelating agent, from about 0.7% to about 20% by weight a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH3.0 to pH7.0; wherein the weight percentages are based on the total weight of the pharmaceutical formulation.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 62 / 853,215, filed May 28, 2019, the entire disclosure of which is incorporated herein by reference. Technical Field The present invention relates generally to broad-spectrum bactericidal formulations for the treatment or prevention of bacterial infections in mammals, including bacterial infections accompanied by viral infections. The present invention also includes methods of treating bacterial infections in mammals by administering a pharmaceutical composition of the present invention to the infected area of ​​the mammal. [Background technology]

[0002] Presented below is background information regarding certain aspects of the invention that may be related to technical features referred to in the detailed description, but that are not necessarily described in detail. That is, certain components of the invention may be described in more detail in the material discussed below. The following discussion should not be construed as an admission regarding the relevance of the information to the claimed invention or the prior art effect of the described material.

[0003] Bacterial resistance to small molecule antibiotic drugs is a growing medical problem that ultimately reduces or eliminates many treatment options for bacterial infections, leaving patients susceptible to previously treatable conditions. Antibacterial small molecule drugs typically work by targeting specific bacterial enzymes, blocking critical biosynthetic pathways necessary for bacteria to grow and withstand internal and external stresses. Bacteria often develop genetic resistance to these drugs by modifying the enzyme target site of the small molecule drug. This resistance is very quickly passed on to bacterial offspring, creating new populations of antibiotic-resistant strains and substrains. The World Health Organization has determined that global antibiotic resistance remains a major threat, even with the development of new antibiotics.

[0004] Furthermore, antibacterial agents are known to cause unintended negative side effects that compromise the health of treated patients (Cunha, Burke A. "Antibiotic side effects" Medical Clinics of North America 85.1 (2001): 149-185). Most side effects associated with antibiotic treatment are not life-threatening. However, these side effects can reduce patient compliance with completing the prescribed treatment course, thereby contributing to bacterial resistance in the global population. For example, commonly prescribed drugs such as tetracyclines often induce photosensitivity in patients, while patients taking beta-lactams often develop fevers or suffer from allergic reactions that can be life-threatening in some cases.

[0005] With regard to viral and bacterial respiratory co-infections, elimination of bacterial populations (both at the onset of viral infection and during the advanced stages of viral infection) is important to avoid or reduce serious respiratory conditions and mortality. Summary of the Invention [Problem to be solved by the invention]

[0006] Thus, there is a critical and unmet need for broad-spectrum antiseptic formulations that can be used to treat or prevent a wide range of diseases of bacterial etiology without the problems often associated with antimicrobial agents. [Means for solving the problem]

[0007] Summary of the invention The following brief summary is not intended to be inclusive of all features and aspects of the present invention, nor is it intended to imply that the present invention must include all features and aspects discussed in this summary.

[0008] The present disclosure overcomes the problems associated with antibiotics by using a universal bactericide formulation containing lysozyme and one or more divalent metal chelators as cofactors to enhance the effectiveness of lysozyme for treating bacterial infections in mammals. The formulations provided herein use lysozyme and excipients that are safe and minimize unintended negative side effects. The present disclosure further includes methods for treating various infectious diseases of bacterial etiology in mammals using lysozyme formulations.

[0009] Lysozyme is the most prominent member of the very large class of glycosidases or glycohydrolases, i.e., enzymes that catalyze the transfer of glycosyl groups to water. Lysozyme catalyzes the hydrolysis of polysaccharide components of the cell walls of Gram-positive bacteria. To do this, lysozyme promotes the cleavage of glycosidic CO bonds between N-acetylmuramic acid and N-acetyl-D-glucosamine residues in the peptidoglycan component of the cell wall. Early crystal structure studies of lysozyme revealed that the enzyme binds to substrates such that the target CO bond falls within two, and only two, potential catalytic groups, Glu35 and Asp52.

[0010] Lysozyme exerts its antibacterial activity by contacting bacteria and destroying their cell walls, which are formed on phospholipid membranes. Bacterial cell walls protect them from osmotic pressure between the inside and outside of the cell, which can induce harmful cellular stress, including lysis. Depending on the structure of their cell walls, bacteria can be classified as Gram-negative or Gram-positive. Lysozyme is known to be a bactericidal agent against most Gram-positive bacteria, but it exhibits weak activity against Gram-negative bacteria. This is primarily due to the fact that Gram-negative bacteria contain lipopolysaccharide (LPS), which comprises the outer membrane covering the peptidoglycan layer found between the outer and inner membranes.

[0011] In the presence of divalent cation chelators such as EDTA, lysozyme becomes as effective against Gram-negative bacteria as it is against Gram-positive bacteria. The mechanism of this effect of chelators is not well understood. However, it is hypothesized that removal of stabilizing divalent cations from the LPS layer by the chelator results in the release of LPS, allowing the molecule to penetrate the outer membrane.

[0012] Lysozyme and EDTA are generally recommended as safe (GRAS) by the FDA. The inventors have discovered that lysozyme as an active ingredient in a therapeutic formulation offers several advantages, including, but not limited to, being easy to manufacture and administer to patients, while also being highly safe and well-tolerated by patients through a variety of administration routes.

[0013] Lysozyme is active over a wide range of pH values, but its optimum pH lies between pH 4 and pH 6. In fact, lysozyme exhibits stable catalytic efficiency throughout this pH range.

[0014] In some embodiments of the present disclosure, lysozyme acts as the primary bactericidal component in the bactericidal formulation by destroying the cell walls of bacteria infecting mucosal membranes or the environment surrounding the infected area. In some embodiments, the bactericidal formulation will contain a chelating agent that enhances the effect of lysozyme against Gram-positive and Gram-negative bacteria. In some embodiments, the bactericidal formulation will contain a pH stabilizer to ensure that 1) the pH of the product, once dissolved in the area of ​​application, is not harmful to the patient; and 2) once the product is dissolved in the area of ​​application, the pH of the resulting solution always remains within the optimal range for lysozyme activity. In yet other embodiments, natural and neutral fillers can be added to dilute the active ingredients of the formulation to the appropriate concentration for local application. In one embodiment, a natural flavor or coloring agent is added to the formulation.

[0015] In some embodiments, the bactericidal formulation is a pharmaceutical powder formulation for administration to a mammal, comprising lysozyme, a pharmaceutically acceptable chelating agent, and a pH-stabilizing salt. The pharmaceutical powder formulation of the present invention can be dissolved in an aqueous solution before administration to a mammal. In some embodiments, the pharmaceutical powder formulation will comprise zinc oxide. In yet other embodiments, the pharmaceutical powder formulation will comprise magnesium citrate.

[0016] In certain embodiments, the antiseptic formulation is in the form of a tablet for oral administration, comprising lysozyme, a pharmaceutically acceptable chelating agent, a pH-stabilizing salt, and a thickener and / or excipient comprising a resin. In one embodiment, the pharmaceutical tablet formulation comprises at least one of a flavoring agent, a coloring agent, or a combination thereof. In yet another aspect, the pharmaceutical tablet formulation is in the form of a chewable tablet for oral administration.

[0017] In certain embodiments of the present invention, the pH-stabilizing salt has a buffering capacity in the range necessary to maintain optimal catalytic efficiency of lysozyme. In a preferred embodiment, the pH-stabilizing salt has a buffering capacity in the range of pH 3.0 to about pH 7.0. More preferably, the pH-stabilizing salt stabilizes the pH of the formulation in solution at a pH of about pH 6.0 to about pH 6.8.

[0018] The inventors have discovered that certain agents that act synergistically with the lytic activity of lysozyme can be included in the formulations of the present disclosure. Thus, in some embodiments of the present disclosure, a disinfectant formulation containing lysozyme will include a synergistic component. In some embodiments, the synergistic component is a solubilizer. In one embodiment, the formulation is in a solid (powder or tablet) state before administration, e.g., for oral use, and a mixture of citric acid and sodium bicarbonate can be used to accelerate its dissolution. In some embodiments, the synergistic component is a desiccant or disinfectant carrier. In one embodiment of a formulation for external (skin) use, the carrier can serve four functions: a) carrier; b) keeping the application site dry; c) disinfectant to maintain the condition of the outer portion of the treatment area; and d) releasing the disinfectant component of the formulation into the interior of the treatment area upon contact with any wet area.

[0019] In some embodiments of the present invention, the pharmaceutical preparation of the present invention can be used to treat bacterial skin infections.For example, the pharmaceutical preparation can be used to treat or prevent infections in skin sores that persist in diabetic and immunologically weakened patients; skin burns (at different levels), which often become infected due to environmental exposure, causing patients to undergo painful recovery procedures and other bacterial skin infections.

[0020] In one embodiment of the present invention, the pharmaceutical preparations disclosed herein can be used to treat bacterial infections in the oropharynx, pylorus, and esophagus tissue regions. Permanent hiding areas for bacteria are the paranasal sinuses, oral cavity (e.g., gums), and throat, causing persistent infections in the mucosal tissues surrounding these regions. Because these regions share a common entrance, the pharmaceutical preparations of the present invention allow for simultaneous treatment of these regions.

[0021] In some embodiments, the pharmaceutical preparation can be used to treat bacterial infections of the large intestine, including acute salmonellosis, colitis, and diverticulitis. In some embodiments, the primary component of the preparation is lysozyme, present in sufficient quantity and administered in a large volume of carrier (such as water) to provide a pharmaceutically effective amount of the enzyme to the large intestine. In a non-limiting embodiment, the preparation includes an agent that induces intestinal flushing, promoting the flow of water into the large intestine and forcing bacteria into the liquid suspension that quickly forms within the intestinal lumen. This allows the lysozyme to easily attack and destroy the bacteria (a laxative effect).

[0022] In certain embodiments, the pharmaceutical formulations can be used to treat bacterial infections present in the tissues of the upper and lower respiratory tract, including the sinuses and lungs. While these two areas share a common entrance, the sinuses often become a latent zone, allowing for reinfection. In another aspect of the present invention, the pharmaceutical formulations provided herein can be used to treat or cure bacterial infections in the bloodstream or vital organs of a mammal. The sinuses, lungs, blood, and vital organs are very delicate and sensitive to insoluble substances. Therefore, some aspects of the present invention provide formulations containing only lysozyme and ethylenediaminetetraacetic acid (EDTA). In some embodiments, sodium bicarbonate is provided together with EDTA in its acid form. In yet other embodiments, the pharmaceutical formulations contain an EDTA salt and a trace amount of sodium bicarbonate to stabilize the pH of the formulation after it is dissolved in aqueous solution.

[0023] In a further aspect, the formulations of the present invention can be used during surgical procedures or during post-surgical recovery, since the irrigant or irrigation solution creates sterility in the surgical intervention area.As a result, the formulations disclosed herein can eliminate the need to use antibiotics during surgery or post-surgical recovery.It will be readily apparent to those skilled in the art that the bactericidal effect of a formulation containing lysozyme during surgery and its appropriate use during post-surgical recovery can eliminate the need to use anti-inflammatory drugs during post-surgical recovery.

[0024] In yet another aspect of the invention, the pharmaceutical preparations can be used to treat bacterial infections of the eye, such as bacterial conjunctivitis.

[0025] The foregoing and other objects, features and advantages of the present invention will become apparent from the following more particular description of the preferred embodiment of the invention, as illustrated in the accompanying drawings, which illustrate the principles of the invention, but the invention of this disclosure is not limited thereto. [Brief explanation of the drawings]

[0026] [Figure 1] FIG. 1 is a graph showing the Silness and Low Gingival Index of patients treated with Lysodent™ versus chlorhexidine. [Figure 2] 2A-B are graphs showing gingivitis-related inflammation from days 1 to 28 in patients treated with Lysodent™ in FIG. 2A and gingivitis-related bleeding from days 1 to 28 in patients treated with Lysodent™ in FIG. 2B. [Figure 3] 3A-B are graphs showing gingivitis-related inflammation from days 1 to 28 in patients treated with chlorhexidine in FIG. 3A and gingivitis-related bleeding from days 1 to 28 in patients treated with chlorhexidine in FIG. 3B. [Figure 4] 4A-B are graphs showing gingivitis-related inflammation observed in patients treated with chlorhexidine in FIG. 4A or Lysodent™ in FIG. 4B. [Figure 5] 5A-B are graphs showing gingivitis-related bleeding observed in patients treated with chlorhexidine in FIG. 5A or Lysodent™ in FIG. 5B. [Figure 6] FIG. 6 is a graph showing pain reported in patients treated with chlorhexidine or Lysodent™ after dental extraction surgery. [Figure 7] FIG. 7 is a graph showing reported inflammation in patients treated with chlorhexidine or Lysodent™ following dental extraction surgery. [Figure 8] FIG. 8 is a graph showing patient response to the taste of chlorhexidine or Lysodent™ formulations following dental extraction surgery. [Figure 9] FIG. 9 is a graph showing patients whose taste perception was altered after being treated with chlorhexidine or Lysodent™ following dental extraction surgery. [Figure 10] FIG. 10 is a graph showing pain reported in patients treated with chlorhexidine or Lysodent™ after dental implant surgery. [Figure 11] FIG. 11 is a graph showing inflammation reported in patients treated with chlorhexidine or Lysodent™ after dental implant surgery. [Figure 12] FIG. 12 is a graph showing reported bleeding in patients treated with chlorhexidine or Lysodent™ after dental implant surgery. [Figure 13] FIG. 13 is a graph showing patient response to the flavor of formulations treated with chlorhexidine or Lysodent™ after dental implant surgery. [Figure 14] FIG. 14 is a graph showing patients whose taste perception was altered after being treated with chlorhexidine or Lysodent™ following dental implant surgery. [Figure 15] FIG. 15 is a schematic diagram of teeth evaluated for Silness and Low Gingival Index. DETAILED DESCRIPTION OF THE INVENTION

[0027] Detailed Description of the Preferred Embodiments The following detailed description includes references to the accompanying drawings, which form a part of the detailed description. The drawings show, by way of illustration, specific embodiments in which the invention may be practiced. The embodiments herein may be combined, other embodiments may be utilized, or changes may be made based on structural, chemical, or other logical changes that fall within the scope of the invention. Therefore, the following detailed description should not be considered limiting in scope.

[0028] In understanding the scope of the present disclosure, the terms "including" and "comprising" and their derivatives, as used herein, are intended to be open-ended terms that specify the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other, unrecited features, elements, components, groups, integers, and / or steps. The same also applies to words of similar meaning, such as "comprise" and "have," and their derivatives. The term "consisting of" and its derivatives, as used herein, is intended to be closed-ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other, unrecited features, elements, components, groups, integers, and / or steps. The term "consisting essentially of" as used herein is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps, as well as things that do not materially affect the basic and novel characteristics of the features, elements, components, groups, integers, and / or steps. It should be understood that reference to any of these transition terms (i.e., "comprising," "consisting of," or "consisting essentially of") directly supports substitution with any of the other transition terms not specifically used. For example, amending the term "comprising" to "consisting essentially of" is directly supported by this definition.

[0029] As used herein, the term "about" means inclusive of the stated value and within an acceptable range of variation of the particular value as determined by one of ordinary skill in the art, taking into account the measurement and the error associated with measuring the particular quantity (i.e., limitations of the measurement system). For example, "about" can mean within one or more standard deviations, or within ±30%, 20%, 10%, or 5% of the stated value.

[0030] Generally, as used herein, the term "or" includes "and / or."

[0031] As used herein, multiple compounds or steps may be presented in a common list for convenience; however, these lists should be construed as though each member of the list were individually identified as a separate and unique member. Accordingly, individual members of such lists should not be construed as de facto equivalents to other members of the same list solely based on their presentation in a common grouping, absent indication to the contrary.

[0032] The terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting. As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms, including "at least one," unless the content clearly dictates otherwise. "At least one" should not be construed as limiting "a" or "an." "Or" means "and / or." As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0033] By "pharmaceutically acceptable chelating agent" is meant an agent that is safe and effective for administration to mammals, including humans, and that produces minimal or no negative side effects, either alone or in combination with other pharmaceutical ingredients.

[0034] A "pharmaceutically acceptable aqueous solution" includes an aqueous solution suitable for administration to a mammal, e.g., a human, including, but not limited to, topical, oral, subcutaneous, or intravenous administration.

[0035] The phrase "a pharmaceutically acceptable amount" of the present invention refers to either the amount of lysozyme as an active agent or the total amount of a pharmaceutical composition containing lysozyme that will treat or cure the bacterial infection at the affected site.

[0036] The "buffering capacity" of a pH-stabilizing salt is the salt's ability to resist a pH change in response to the addition of a strong acid or strong base in solution. For example, buffering capacity can be measured according to the amount of strong acid or strong base required to change the pH of 1 liter of solution by 1 pH unit under standard temperature and pressure conditions. Empirical methods for determining the buffering capacity of a given salt by both acid titration and base titration over a given range of pH change from the desired pH of the composition include conventional techniques well known in the art.

[0037] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. Furthermore, terms as defined in commonly used dictionaries should be interpreted to have a meaning consistent with their meaning in the context of the relevant art and this disclosure, and are not to be interpreted as idealized or overly formal unless expressly defined as such herein.

[0038] The inventors of the present disclosure have surprisingly demonstrated that the formulations comprising lysozyme provided herein act as universal bactericides, killing both Gram-negative and Gram-positive bacteria. The formulations of the present disclosure do not cause adverse side effects on human tissues and organs, even during long-term use.

[0039] As a non-limiting example, the inventors have found that a suitable formulation according to the present invention is a 50 mg powder formulation. In some embodiments, the amount of lysozyme ranges from about 1 mg to about 45 mg. In some aspects of the present invention, the amount of lysozyme is greater than about 10 mg, or from about 20 mg to about 40 mg, or optimally about 35.6 mg of lysozyme. In some aspects of the present invention, the formulation contains about 1 mg to about 45 mg of chelating agent. In other aspects, the amount of chelating agent is about 5 mg to about 20 mg, or from about 10 mg to about 20 mg, or optimally about 12.2 mg of chelating agent. In some embodiments, the amount of pH stabilizing salt is about 0.1 mg to about 10 mg. In some aspects of the present invention, the amount of pH stabilizing salt is about 0.5 mg to about 5.0 mg, or from about 1.0 mg to about 2.0 mg, or optimally about 1.3 mg of pH stabilizing salt. In some embodiments, 100 mg of a powder formulation according to the present invention (twice the amount above, respectively) is dissolved in 10 ml of aqueous solution. In other embodiments, 250-300 mg of a powder formulation according to the present invention (five and six times the amount above, respectively) is dissolved in 1.0 liter of aqueous solution. In still other aspects of the present invention, as described above, 300, 400, 500, 700, or 1000 mg of powder formulation per liter of aqueous solution can be used.

[0040] In some embodiments, the 50 mg powder formulation described above comprises zinc oxide in a weight ratio of about 1:99 to about 50:50 of total lysozyme formulation (e.g., 50 mg of lysozyme powder formulation) to zinc oxide. In other aspects, the ratio of lysozyme formulation to zinc oxide is about 5:95 to about 20:80, and in some embodiments of the invention, the amount of lysozyme formulation to zinc oxide is about 10:90 to about 30:70.

[0041] In a further non-limiting example, the inventors have found that a suitable formulation according to the present invention is a 500 mg tablet formulation. In some embodiments, the amount of lysozyme ranges from about 1 mg to about 100 mg. In some aspects of the present invention, the amount of lysozyme is about 10 mg to about 50 mg, or about 30 mg to about 40 mg, or optimally about 35.6 mg. In some aspects of the present invention, the formulation contains about 1 mg to about 50 mg of chelating agent. In other aspects, the amount of chelating agent is about 5 mg to about 40 mg, or about 10 mg to about 20 mg, or optimally about 12.2 mg of chelating agent. In still other aspects, the amount of pH stabilizing salt is about 1 mg to about 10 mg. In certain aspects, the amount of pH stabilizing salt is about 2.5 mg to about 5 mg, or about 3.5 mg to about 4.5 mg, or optimally about 4.0 mg of pH stabilizing salt. Additionally, some embodiments of the formulation include about 10 mg to about 450 mg of a pharmaceutically acceptable resin. In other embodiments, the formulation includes about 100 mg to about 400 mg, or about 200 mg to about 390 mg, or about 446.6 mg of a pharmaceutically acceptable resin. In some embodiments, the formulation includes about 0.1 mg to about 10 mg of a flavoring agent and / or, separately, an equal or different amount of a coloring agent. Optionally, the formulation includes about 0.3 mg to about 0.5 mg of a flavoring agent and / or an equal or different amount of a coloring agent. In embodiments of the present invention, the tablet is a chewable tablet.

[0042] In a preferred embodiment, the pharmaceutical formulation is a powder containing lysozyme and a chelating agent. The inventors of the present disclosure have found that lysozyme is safe and effective in treating a wide range of bacterial infections when present in a relatively low concentration in solution. In some embodiments, the amount of lysozyme is about 0.2% to about 90% by weight, or about 2% to about 80% by weight, or about 6% to about 60% by weight, or about 8% to about 40% by weight, or about 10% to about 20% by weight, or about 12% to about 15% by weight, based on the total weight of the pharmaceutical formulation. In some embodiments, lysozyme is present in an amount of about 73% by weight or less based on the total weight of the pharmaceutical formulation. The present disclosure is not limited by the specific amounts of lysozyme described herein; any amount within the aforementioned ranges can be used.

[0043] Pharmaceutically acceptable chelating agents may be selected from ethylenediaminetetraacetic acid (EDTA) salts, citrate salts, alginate salts, and combinations thereof. The inventors have discovered that the amount of chelating agent useful for providing broad-spectrum antimicrobial activity of the formulations of the present disclosure can be present in a wide range of concentrations. In some embodiments, the amount of chelating agent is from about 0.2% to about 90% by weight, from about 2% to about 40% by weight, from about 4% to about 24% by weight, from about 10% to about 20% by weight, or from about 12% to about 15% by weight, based on the total weight of the pharmaceutical formulation.

[0044] In an embodiment of the present invention, the formulation further comprises a pharmaceutically acceptable pH-stabilizing salt. The pH-stabilizing salt is selected from the group consisting of citrate and sodium bicarbonate. The inventors have found that a useful amount of pH-stabilizing salt for maintaining a safe and effective pH when the formulation is dissolved in an aqueous solution is about 0.2% to about 20% by weight. In some embodiments, the amount of stabilizing salt may be present in an amount of about 0.5% to about 4% by weight, about 0.7% to about 2.6% by weight, about 0.8% to about 2% by weight, or about 0.9% to about 1.0% by weight, based on the total weight of the pharmaceutical formulation.

[0045] In one aspect of the present invention, the formulation includes a resin. Non-limiting examples of resins useful in the formulations of the present disclosure include plant-derived resins. A preferred resin for the formulation is alginate resin. In some embodiments, the resin is present in an amount of 2% to about 90% by weight. In other embodiments, the resin is present in an amount of about 20% to about 89% by weight, about 40% to about 80% by weight, or about 50% to about 70% by weight, based on the total weight of the pharmaceutical formulation. In some aspects of the present invention, the resin is present in an amount of about 78% by weight, based on the total weight of the pharmaceutical formulation.

[0046] In some aspects of the invention, the formulation further comprises magnesium citrate. In certain embodiments, the magnesium citrate is present in an amount of about 0.1 grams to about 5.0 grams. In other aspects of the invention, the magnesium citrate is present in the formulation in an amount of about 0.5 grams to about 2.5 grams, or about 1.0 grams to about 2.0 grams.

[0047] In another embodiment of the present invention, the formulation includes a colorant, a flavoring agent, or a combination thereof. The colorant and / or flavoring agent is present in an amount of about 0.02% by weight to about 2% by weight, based on the total weight of the pharmaceutical formulation, in equal or different amounts. In another embodiment of the present invention, the colorant and / or flavoring agent is present in an amount of about 0.06% by weight to about 1% by weight, based on the total weight of the pharmaceutical formulation, in equal or different amounts.

[0048] The pharmaceutical formulations of the present disclosure can be dissolved in an aqueous solution and then administered to a mammal. In some embodiments, the formulations are dissolved in water, or sterile water and / or deionized water. In other embodiments, the formulations are dissolved in saline.

[0049] The present disclosure provides a method for treating a bacterial infection in the gastrointestinal tract of a mammal, comprising orally administering (i.e., ingesting) a pharmaceutical powder formulation or effervescent tablet dissolved in water, wherein the powder formulation comprises: (i) lysozyme, (ii) magnesium citrate, (iii) a pharmaceutically acceptable chelating agent, and (iv) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. In a non-limiting embodiment, the formulation is dissolved in about 500 mL to about 700 mL of water. In some embodiments, the citric acid and sodium bicarbonate are present in stoichiometric amounts. Preferably, the method comprises administering the entire solution to the patient at one time. In some embodiments of this method, a second dose is administered 8 hours after the first dose.

[0050] The present disclosure provides a method for treating bacterial infections of the oral cavity of a mammal, comprising orally administering a chewable pharmaceutical tablet, wherein the tablet comprises: (i) lysozyme, (ii) a pharmaceutically acceptable resin, (iii) a pharmaceutically acceptable chelating agent, (iv) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0, and, optionally, a colorant, a flavoring agent, or a combination thereof. The resin thickener is added to achieve a viscous mixture in the mouth when the tablet is chewed, so that when swallowed, the solution moves slowly through the throat and esophagus, resulting in a longer residence time in these areas and a formulation that is more effective in killing bacteria encountered on its way to the stomach. In some embodiments, a stoichiometric mixture of citric acid and sodium bicarbonate is added in an amount of about 5% by weight, based on the total weight of the formulation, to promote dissolution of the solid tablet upon contact with the patient's saliva. In some aspects of this method, the formulation is administered once every 12 hours.

[0051] The present disclosure further provides a method for treating a bacterial respiratory tract infection in a mammal, comprising introducing a pharmaceutical formulation into the respiratory tract of the mammal, wherein the pharmaceutical formulation is an aqueous solution administered using a nebulizer and comprises: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, and (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. In some embodiments, 50 to 100 mg of the formulation is dissolved in 5.0 mL of an aqueous solution containing saline or distilled water. In some embodiments, a nebulizer is used to treat the lungs of an infected mammal every 8 hours until the composition is applied three times.

[0052] Another aspect of the present invention is a method of treating sepsis in a mammal comprising introducing a pharmaceutical formulation into the bloodstream of the mammal, wherein the pharmaceutical formulation comprises intravenously administered saline solution comprising: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, and (iii) a pharmaceutically acceptable pH stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. In some embodiments, the saline solution comprises about 100 mg to about 1000 mg of the pharmaceutical composition per liter of solution.

[0053] The present disclosure further provides a method of treating a bacterial skin infection in a mammal, comprising contacting an affected area of ​​skin with saline, wherein the saline comprises: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, and (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. In some embodiments of the present invention, the method further comprises drying the affected area of ​​skin; and applying a pharmaceutical powder formulation to the affected area of ​​skin, wherein the formulation comprises: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0, and (iv) zinc oxide.

[0054] Provided herein are methods for treating or preventing bacterial infections in a mammal undergoing a surgical procedure, comprising contacting the area of ​​surgical intervention with saline, wherein the saline comprises: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, and (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. In some embodiments, the method further comprises applying the saline to a suture site after the surgical procedure. In some embodiments, the method further comprises applying a pharmaceutical powder formulation to an affected area of ​​skin after the surgical procedure, wherein the formulation comprises: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0, and (iv) zinc oxide.

[0055] Examples of bacterial infections that can be treated using the compounds of the present invention include, but are not limited to, Atopobium (e.g., parvum, limae), Bacillus (e.g., Bacillus anthracis), Bacteroides (e.g., fragilis), Bordetella (e.g., pertussis), Borrelia burgdorferi, Bradydia extorctas, Campylobacter (e.g., jejuni), Catonella morbi, Centipede peridontii, Chlamydia (e.g., C. trachomatis), Clostridium (e.g., C. difficile, C. haschiforme, C. histolyticum, C. perfringens, C. subterminale, C. sporogenes, C. bifementans, C. botulinum, C. oedematiens, C. perfringens, C. tetani), Cryptobacterium curtum, C. pneumocystis, E. coli, E. sulci, C. filifactorii, C. allos, Fusobacterium (e.g., C. periodonticum, nucleatum), Granulicatella asiacens, Haemophilus influenzae, Lactobacillus acidophilus, Listeria monocytogenes (e.g., Monocytogenes), Mollusca bacillus (e.g., Thymidum, Bescum), Mycobacterium (e.g., Tuberculosis), Neisseria (e.g., Gonorrhea), Prevotella, Porphyromonas (Endodontalis, Gingivalis), Streptococcus pneumoniae, Pseudoramibacter aractoticus, Salmonella enterica, Selenomonas sputigena, Shigella, S. exigua, Staphylococcus epidermidis (e.g., Staphylococcus aureus [MRSA], S. epidermidis), Streptococcus (e.g., Pneumoniae, Mitutos, Oralis, Salivarylus, Sanguinis, Milleri, Mutans, Sobrinus, Anginosus), Tannerella forsythia, Treponema (e.g., Denticola, Socranskii, Pallidum, Pectinovorum, Amylovorum, Medullaria), Vibrio (e.g., Cholera).

[0056] Provided herein is a method for treating or preventing a bacterial infection in a mammal that manifests as a superinfection with a virus, comprising contacting the affected area with saline, wherein the saline contains: (i) lysozyme, (ii) a pharmaceutically acceptable chelating agent, and (iii) a pharmaceutically acceptable pH-stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0.

[0057] In one embodiment of the present invention, the compounds of the present invention can be used as a therapeutic agent for bacterial infections accompanied by any viral infection. In one preferred embodiment, the virus is one that causes a respiratory infection. Examples of viruses that cause respiratory infections include, but are not limited to, influenza, paramyxoviruses (e.g., respiratory syncytial virus, parainfluenza, metapneumovirus), picornaviruses (enteroviruses, rhinoviruses), coronaviruses (229E, NL63, OC43, HKU1, MERS-CoV, SARS-CoV, SARS-CoV-2 [which causes COVID-19 syndrome]), adenoviruses, and parvoviruses.

[0058] When the compositions of the present invention are administered at the onset of a viral infection, the complex interactions between viruses and bacteria that can challenge the immune system (which increase the morbidity and mortality of viral infections) are avoided or minimized. Furthermore, during the plateau period of a viral infection, continued administration of the compositions periodically (e.g., once every 48 hours) after the initial shock treatment (e.g., three times, once every 8 hours) avoids viral-bacterial superinfection; this gives the immune system a chance to win the battle against the virus.

[0059] When the compositions of the present invention are administered as shock therapy (e.g., three times, once every eight hours) after the onset of symptoms or during the critical period of a viral infection (when viral-bacterial superinfection has already begun), the synergistic action of the virus and bacteria is stopped, avoiding runaway immune system and further systemic and microanatomical damage; this reduces morbidity and promotes patient recovery.

[0060] The compounds of the present invention can be used as a treatment for all bacterial infections, whether or not accompanied by a viral infection. However, in one embodiment, the compounds of the present invention can be used as a treatment for bacterial infections accompanied by a viral infection as a means of reducing the acuity of disease and mortality by avoiding, interfering with, or stopping the synergistic partnership between viruses and bacteria that overwhelms the immune response.

[0061] More particularly, the present invention is described by the following items, which represent preferred embodiments thereof:

[0062] 1. about 2% by weight to about 80% by weight of lysozyme; about 2% to about 40% by weight of a pharmaceutically acceptable chelating agent; 1. A pharmaceutical formulation for administration to a mammal, comprising about 0.7% to about 20% by weight of a pharmaceutically acceptable pH stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0; said weight percentage being a percentage of the total weight of the pharmaceutical formulation. 2. The pharmaceutical formulation according to item 1, wherein the formulation is dissolved in a pharmaceutically acceptable aqueous solution before administration to a mammal. 3. The pharmaceutical formulation according to item 1, wherein the chelating agent is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA) salts, citrate salts, alginate salts, and combinations thereof. 4. The pharmaceutical formulation according to item 1, wherein the pH stabilizing salt is selected from the group consisting of citrate and sodium bicarbonate. 5. The pharmaceutical formulation according to item 1, wherein the formulation further comprises a resin. 6. The pharmaceutical formulation according to item 1, wherein the formulation further comprises zinc oxide. 7. The pharmaceutical formulation according to item 1, wherein the formulation further comprises magnesium citrate. 8. The pharmaceutical formulation according to item 5, wherein the formulation comprises a chewable tablet form. 9. The pharmaceutical formulation according to item 2, wherein the formulation is provided in a nebulizer. 10. The pharmaceutical formulation according to item 5, wherein the formulation comprises a colorant. 11. The pharmaceutical formulation according to item 5, wherein the formulation comprises a flavoring agent. 12. The pharmaceutical formulation according to item 1, wherein the lysozyme is present in an amount of 80% by weight or less, based on the total weight of the pharmaceutical formulation. 13.i. about 2% to about 80% by weight of lysozyme; ii. about 2% to about 40% by weight of a pharmaceutically acceptable chelating agent; iii. A method for treating or preventing a bacterial infection in a mammal, comprising administering to the infected area of ​​the mammal a pharmaceutical formulation comprising about 0.7% by weight to about 20% by weight of a pharmaceutically acceptable pH stabilizing salt having a buffering capacity in the range of pH 3.0 to pH 7.0. 14. The method according to item 13, wherein the infection is a skin infection. 15. The method according to item 14, wherein the composition is in saline. 16. The method according to item 13, wherein the composition is a pharmaceutical powder formulation. 17. The method of claim 16, wherein the composition further comprises zinc oxide. 18. The method according to item 17, wherein the skin is dried before administering the composition. 19. The method according to item 13, wherein the infectious disease is a respiratory infection. 20. The method of item 19, wherein the composition is administered with a nebulizer. 21. The method according to item 13, wherein the composition is dissolved in water. 22. The method according to item 21, wherein the infection is a gastrointestinal infection. 23. The method of claim 22, wherein the composition further comprises magnesium citrate. 24. The method according to item 21, wherein the infection is in the large intestine. 25. The method according to item 21, wherein the infection is in the oropharyngeal mucosa. 26. The method according to item 13, wherein the infection is in the oral cavity. 27. The method according to item 26, wherein the composition is administered orally. 28. The method according to item 27, wherein the composition is in the form of a chewable pharmaceutical tablet. 29. The method according to item 28, wherein the composition further comprises a pharmaceutically acceptable resin. 30. The method according to item 15, wherein the saline solution contains about 100 mg to about 300 mg of the composition per liter of solution. 31. The method of item 15, wherein saline is introduced into the bloodstream. 32. The method according to item 31, wherein the treatment is for sepsis. 33. The method according to item 15, wherein the infection is in one or more vital organs. 34. The method according to item 15, wherein the solution is introduced into the surgical site during a surgical procedure. 35. The method according to item 21, wherein the infection is in the eye. 36. The method according to item 35, wherein the infectious disease is conjunctivitis. 37. The method according to item 13, wherein the bacterial infection is accompanied by a viral infection. 38. The method according to item 37, wherein the viral infection comprises a respiratory infection. 39. The method according to item 38, wherein the virus causing the viral infection is influenza, coronavirus, adenovirus, or parvovirus. 40. The method according to item 39, wherein the virus is a coronavirus. 41. The method of item 40, wherein the virus is MERS-CoV, SARS-CoV, or SARS-CoV-2. 42. The method according to item 41, wherein the virus is SARS-CoV-2. 43. The method according to item 42, wherein the virus causes COVID-19.

[0063] The compositions and processes of the present invention will be better understood in connection with the following examples, which are intended as illustrative only and not as limiting the scope of the invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications, including but not limited to those relating to the processes, formulations and / or methods of the present invention, can be made without departing from the true spirit of the invention and the scope of the appended claims. Example [Example]

[0064] An example was developed to compare the effectiveness of a mouthwash formulated according to the present invention (Lysodent™) with a comparative example (digluconate 0.12%, chlorhexidine) in dental surgery and periodontal disease. Lysodent™ was used alone in each study. Chlorhexidine was used as a mouthwash during surgery, and Chlorhexidine Plus (chlorhexidine + analgesic + antibiotic + anti-inflammatory) was used during post-operative recovery follow-up.

[0065] The formulation of Lysodent™ is shown in Table 1. [Table 1] [Table 1]

[0066] Study population Seventy subjects (gingivitis, n = 40; surgery, n = 30) were invited to participate in this clinical trial. The subjects were selected in a way that allowed for comparability and fair analysis. Clinical examiners divided the subjects into two groups according to their diagnosis: gingivitis or surgery candidates. Table 2 shows the inclusion criteria for the study population. [Table 2] [Table 2]

[0067] Patient recruitment and informed consent process The study was approved by the Research Ethics Committee (ETIKOS) on June 21, 2018. Patients currently attending the EIBEGO Department of Surgery and Periodontology were invited to be part of the study. Subjects were given an informed consent form, and investigators were available to answer any questions they may have had. Only after subjects fully understood the benefits and risks of the study were they asked to sign the informed consent form, and a copy was given to them for their records. Subjects were also informed of their right to withdraw from the study at any time they wished, without affecting their future treatment at the EIBEGO clinic.

[0068] Baseline, Visit 1 After the informed consent process, medical and dental histories were obtained. A comprehensive oral examination, including the Low-Silness Gingivitis Index and X-rays, was performed. Subjects were then randomly assigned to one of two treatment groups: Lysodent™ or chlorhexidine. Subjects receiving Lysodent™ were instructed to combine 50 mg of Lysodent™ (pre-packaged in an envelope) with water to create a mouthwash using the pre-measured dose cup included in the product kit. Subjects were asked to rinse twice daily for one minute for 28 days: once in the morning after breakfast and brushing, once in the evening, and once before bedtime. Subjects receiving chlorhexidine were instructed to use the pre-measured dose cup included in the product kit to rinse 15 ml of the mouthwash three times daily for one minute. They were asked to rinse in the morning before breakfast and brushing, again 8 hours later, and again before bedtime.

[0069] Visit 2 - 3 days after using the product Compliance with product use was assessed, and subjects were reinstructed as needed. Subjects were evaluated using a comprehensive oral examination, including the Low-Silness Gingivitis Index. A survey was then administered to record pain level (if any), comments regarding flavor, and overall product perception. Subjects were asked to continue using the product at home as directed.

[0070] Visit 3 - 7 days after investigation and product use Photographs of the surgical site were then taken, and a survey was administered to record pain levels (if any), comments about flavor, and overall product perceptions. Subjects were instructed to discontinue use of the product after this visit.

[0071] Visit 4 - 14 days after investigation and product use Photographs of the surgical site were then taken and a survey was administered to record pain level (if any), comments regarding flavor, and overall product perception. This was the final visit and subjects were dismissed from the study.

[0072] Research method Low-Silness Gingivitis Index Measurement of oral hygiene status by the Low-Silness Gingivitis Index is based on recording both soft debris and mineralized deposits on the teeth, as shown in Figure 15. Missing teeth are not replaced. A score of 0 to 3 is assigned to each of the four tooth surfaces (buccal, lingual, mesial, and distal). The four scores are added and divided by 4 to assign a dental plaque index, with the following scores and criteria shown in Table 3: [Table 3] [Table 3]

[0073] Research results Gingivitis Group Comparison of clinical dental prophylaxis and post-treatment outcomes in patients with gingivitis treated with either Lysodent™ or chlorhexidine mouthrinse for 14 days. Lysodent™ was significantly more effective in treating gingivitis. At Visit 2 (3 days of Lysodent™ treatment), subjects had a GI (Low-Silness Index) score of less than 0.5, classifying them as healthy from a periodontal (gingivitis) perspective (Figure 1). At Visit 4 (14 days of Lysodent™ treatment), no bleeding sites were reported, and gingival inflammation was minimized in 75% of cases. At the end of the study, all subjects had a GI score of zero or approaching zero.

[0074] In contrast to Lysodent™ (Figures 2A and 2B), chlorhexidine treatment showed a much slower recovery in terms of eliminating gingival inflammation and bleeding (Figures 3A and 3B). At the end of the study (28 days of treatment), nearly 50% of chlorhexidine-treated subjects still had a GI score higher than 0.5 (Figure 1). Patients treated with chlorhexidine had an increased number of bleeding and inflamed teeth throughout the 28-day study (Figures 4A and 5A) compared to patients treated with Lysodent™ (Figures 4B and 5B).

[0075] Tooth Extraction Surgery Group Lysodent™ was more effective than Chlorhexidine Plus in controlling pain associated with dental surgery. Only one Lysodent™-treated case (7%) reported mild pain on the first day after starting the medication, in contrast to the reported number of Chlorhexidine Plus cases (75% on the first day, 63% on the second day, and 38% on the third day) (Figure 6).

[0076] Although no cases of inflammation were reported after treatment initiation, Lysodent™ proved superior in preventing inflammation, as 50% of patients using Chlorhexidine Plus reported inflammation during the first 3 days (Figure 7).

[0077] The effectiveness of the treatment in preventing bleeding could not be assessed in the study. No patients reported bleeding during the study.

[0078] Lysodent™ proved superior in patient ratings of product flavor: only 14% of patients using Lysodent™ reported an unpleasant taste in the mouthrinse, while 88% of patients using Chlorhexidine Plus reported an unpleasant flavor (Figure 8).

[0079] Lysodent™ did not affect flavor perception in any case, but 50% of the cases using Chlorhexidine Plus reported a change in food flavor perception (bitter mouth) after 7 days of use (Figure 9).

[0080] Dental Implant Surgery Group Lysodent™ proved to be much more effective at controlling pain than the Chlorhexidine Plus combination: none of the Lysodent™-treated cases (0%) reported pain after starting the medication, compared with 86% of Chlorhexidine Plus patients on day 1, 71% on day 2, 57% on day 3, and 14% on day 14 after starting the medication (Figure 10).

[0081] Lysodent™ proved superior in preventing inflammation, with only one patient (7%) reporting mild irritation from days 1 to 3 after starting treatment, whereas 100% of Chlorhexidine Plus users reported irritation during the same period (Figure 11).

[0082] Lysodent™ also proved superior in preventing bleeding, as no patients reported bleeding, whereas 40% of patients using Chlorhexidine Plus reported bleeding during the first 3 days of the study (Figure 12).

[0083] Patients' ratings of product flavor were also superior for Lysodent™: No patients using Lysodent™ reported an unpleasant taste in the mouthrinse, while 86% of patients using Chlorhexidine Plus reported an unpleasant taste (Figure 13).

[0084] Furthermore, Lysodent™ did not affect flavor perception in any case, whereas 71% of cases using Chlorhexidine Plus reported a change in food flavor perception after 7 days of use (Figure 14).

[0085] The results of the study demonstrate the effectiveness of Lysodent™ in eliminating the presence of bacteria in the oral cavity during surgery and post-operative recovery. Treatment with Lysodent™ was found to prevent symptoms (pain, inflammation, and bleeding) associated with infection or increased bacterial counts in the oral cavity during and after surgery. All results point to the conclusion that Lysodent™ is safe and highly effective when used as an antiseptic during surgery and post-operative follow-up to avoid infection of the surgical area.

[0086] Furthermore, the results of the studies described herein demonstrate that the use of Lysodent™ as an antiseptic during and after surgery results in rapid and efficient recovery and healing of oral tissues. This effect is due to the near-complete sterility created by cleaning during surgery and use of the mouthwash during recovery.

[0087] It is also noteworthy that no adverse events have been reported with the use of Lysodent™. None of its ingredients have been found to be harmful to human tissues or metabolism. As a result, this study provides evidence that Lysodent™ and the formulations disclosed herein can be used as a mouthwash and then swallowed. It is important to note that postoperative analgesics and antibiotics are unnecessary due to the effectiveness of Lysodent™ in eliminating the source of infection. [Example]

[0088] For acute or chronic bacterial infections of the sinuses and lungs LYSIBIOTIC is one of the compositions of the present invention. LYSIBIOTIC is not harmful to the mucosal surface of the lungs and can be applied to any bacterial lung infection, but this application is important when the bacterial strain involved is antibiotic-resistant or the patient is antibiotic-sensitive. LYSIBIOTIC has been used in patients with chronic and very acute lung infections, resulting in complete recovery within 48 hours; moreover, LYSIBIOTIC is very useful in eliminating bacterial superinfections in viral respiratory infections. Because it does not cause any side effects, it is ideal for immunocompromised patients or patients undergoing chemotherapy.

[0089] Instructions for use The most effective way to deliver LYSIBIOTIC formulations to the lungs is via a nebulizer. Dissolving a single dose of LYSIBIOTIC (100 mg) in distilled water or saline has proven highly effective (complete recovery achieved in 100% of cases). The application procedure is as follows: Dissolve 100 mg of the product in 5 ml of distilled water or saline and apply with a nebulizer; repeat the application every 8 hours until three applications have been made. In most cases, it is not necessary to apply the formulation more than three times.

[0090] Protocol execution steps Eligibility criteria: Eligible to participate in the study are patients who meet the following characteristics: 1. Patients aged 15 years or older, regardless of sex, race, or religion, who have been diagnosed with COVID-19 caused by SARS-CoV-2 and hospitalized based on the criteria cited below. 2. Positive COVID-19 test (either rapid / fast test or RT-PCR). 3. Anyone with an acute respiratory illness who has the sudden onset of at least one of the following symptoms: a.Cough b.Sore throat c. Difficulty breathing or d. Fever (subjective), 38°C or higher. 4. Patient is admitted with severe pneumonia of unknown cause and / or evidence of imaging damage corresponding to pneumonic infiltrates (new and / or persistent alveolar infiltrates) and / or signs and symptoms of pulmonary cirrhosis. 5. Written and signed consent by the patient or family member responsible for the use of lysibioyic acid; informed of the characteristics of the study (in compliance with all ethical guidelines, including those of the Helsinki Protocol) and agreeing to voluntarily participate in it.

[0091] Exclusion criteria: 1. Patients requesting exemption from drug use. 2. Patients whose condition makes it appropriate for the physician to suspend their participation in the study (primarily due to the dosage form of the drug Lysibiotic). 3. Hypersensitivity to medications (none reported).

[0092] Written consent of the patient or responsible family member: The patient or responsible family member will be fully informed about all aspects related to the lysibiotic procedure (including aspects of the Helsinki Protocol). The procedure will be tested for its effectiveness in treating general pneumonia and COVID-19 pneumonia. The authorized patient or family member will be asked to read and sign the consent document if they believe they consent to subjecting the patient to the lysibiotic procedure.

[0093] The study will be conducted in two groups: Group A, the standard group for comparison. This group will follow the standard treatment used by the hospital for these patients. Group B, the group under study. This group will follow the standard hospital treatment (excluding the use of antibiotics) minus antibiotics and adding lysibiotic treatment.

[0094] Bioanalytical Analysis and Imaging: Before starting the procedure, the patient will be evaluated with bioanalytical analysis and imaging tests to determine the following parameters: 1) Complete blood count 2) Blood sugar 3) Erythrocyte Sedimentation Rate (ESR) 4) HbA1c 5) Renal profile: a.Urine test b.Urea c. creatinine d.Uric acid 6) Liver profile: Alanine aminotransferase (ALT) - formerly known as SGPT b. Aspartate aminotransferase (AST) - formerly known as SGOT C. bilirubin 7) Thyroid Profile: a. Triiodothyronine (T3) b. Thyroxine (T4-free) C. Thyroid-stimulating hormone (TSH) d. Thyroglobulin 8) Lipid profile: Cholesterol B. Triglycerides C. High-density lipoprotein (cholesterol HDL) d. Low-density lipoprotein (cholesterol LDL) e. Very low density lipoprotein (cholesterol VLDL) 9) PCR 10) PCT 11) Electrolytes 12) CPK 13) Coagulation profile 14) Ferritin 15) Dimer D 16) ELISA using VIH 17) Blood culture 18) SAMR nasal swab 19) EKG 20)DHL 21) Chest PA radiography or TAC(HD) Thorax 22) Arterial gas

[0095] Treatment (Group B) will be applied as follows: a. Dissolve 100 mg (100 milligrams) of Lysibiotic in powder in 5 ml (5 milliliters) of saline, shake gently until completely dissolved, and place in the nebulizer container. b. Place the patient in the nebulizer and turn it on. Ensure the patient remains nebulized until the container is empty. c. Eight hours after the initial spraying, repeat spraying steps a) and b). d. Eight hours after the second spray, repeat spraying steps a) and b). e. Wait 8 h and repeat step IV (bioanalysis analysis and imaging). f. 48 hours after the last spray, repeat spraying steps a) and b). g. Wait 8 h and repeat step IV (bioanalysis analysis and imaging). h. 48 hours after the last spraying, repeat spraying steps a) and b). i. Wait 8 h and repeat step IV (bioanalysis analysis and imaging). j. 48 hours after the last spraying, repeat spraying steps a) and b). k. Wait 8 h and repeat step IV (bioanalysis analysis and imaging).

[0096] From step k), depending on the results of the k-step indicators, the patient will be tracked and treated based on the hospital's COVID-19 management protocol.

[0097] Nebulization of COVID-19 patients should be performed in a negative pressure room under strict nebulization control, ideally using an adult Capacete or Hood Cephalic Chamber to provide full protection from the physician while performing the procedure, and preferably using an FFP3 respirator, or N-95, which has a maximum filter efficiency of approximately 98% and a maximum total leakage rate of 2%.

[0098] Group B continues the same treatment as Group A, except that they follow lysibiotic treatment, but no antibiotics are provided.

[0099] Group A will follow the hospital's standard of care for COVID-19 patients.

[0100] During the study, clinical, bioanalytical, and imaging data from patients in both Groups A and B will be compared according to their daily progress. All parameters of the clinical evaluation obtained from steps e), g), i), and k) of Step V will be analyzed and compared with the parameters of Step IV.

[0101] The patent and scientific literature referenced herein establishes knowledge available to those skilled in the art. U.S. patents and published or unpublished U.S. patent applications cited herein are incorporated by reference. Published foreign patents and patent applications cited herein are incorporated by reference. All other published references, documents, manuscripts, and scientific literature cited herein are incorporated by reference.

[0102] While the present invention has been particularly shown and described with reference to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details can be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

1. (a) Lysozyme; (b) pharmaceutically acceptable chelating agents capable of removing stabilized divalent cations from bacterial membranes; and (c) pharmaceutically acceptable pH-stabilizing components; A pharmaceutical preparation containing, The formulation is structurally configured such that membrane destabilization by a chelating agent is an essential condition for the antibacterial activity of lysozyme, and A pharmaceutical formulation in which the formulation is provided in a non-aerosol dosage form.

2. The pharmaceutical formulation according to claim 1, provided as a solid dosage form selected from tablets, chewable tablets, capsules, powders, or granules.

3. The pharmaceutical formulation according to claim 1, provided as a reconstituted composition configured to dissolve in a pharmaceutically acceptable aqueous medium before administration.

4. A pharmaceutical preparation according to any one of claims 1 to 3, configured for administration by a route selected from oral, topical, systemic, or topical surgical administration.

5. Use of a pharmaceutical preparation according to any one of claims 1 to 4 for manufacturing a pharmaceutical product for controlling a bacterial infection, The antibacterial effect is achieved by enabling the activity of lysozyme through membrane destabilization induced by chelating agents.

6. The use according to claim 5, wherein the bacterial infection involves Gram-negative bacteria that exhibit inherent resistance to lysozyme in the absence of membrane destabilization.

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