Hypochlorite solution for scar tissue treatment

Abstract

The present invention provides a hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.015% to 0.06% by weight (about 150 ppm to 600 ppm by weight), having a pH of 7.5 or greater, and further comprising 0.2% to 3.0% by weight of sodium chloride.

Description

[Technical Field]

[0001] The present invention relates to a hypochlorite solution for the prophylactic or therapeutic treatment of scar tissue in mammals. [Background technology]

[0002] Scar tissue is fibrous tissue that forms after trauma to the skin as part of the healing process or as a result of certain diseases or pathologies. Scar tissue is typically formed from collagen and has several differences compared to normal healthy tissue. For example, scar tissue on the surface of the skin can be characterized by one or more of the following: raised areas, discoloration, increased sensitivity to sunlight, tightness / reduced elasticity, lack of sweat glands, and lack of hair follicles, compared to the normal skin tissue surrounding the scar. Scar tissue can also form internally within the body after severe trauma (e.g., internal scarring after surgery).

[0003] Because the collagen proteins formed during the healing process are usually unidirectional, scar tissue is typically less flexible than normal tissue. This reduces the elasticity of the tissue, which can result in patients experiencing recurrent pain and / or limited normal range of motion. From a cosmetic standpoint, superficial scar tissue is generally considered unsightly.

[0004] Although scar tissue formation is a common part of the healing process, it is necessary to mitigate the effects of scar tissue. Skin wounds are typically sutured after trauma to realign the skin as naturally as possible and to reduce or eliminate the size of the scar. When skin grafts are performed, this may not be possible, resulting in more extensive scarring.

[0005] According to NHS England, the majority of available treatments for scar tissue aim to improve the appearance of scars that have already formed or are in the later stages of healing. For example, cosmetics, steroids, laser therapy, cryotherapy, dermal fillers, skin needling, surgery, and topical creams are used to improve the appearance of scars. For topical applications, creams and oils such as BioOil™ are used to improve the appearance of already formed scars and stretch marks. Such creams and oils are not suitable for application to open wounds in the early stages of the healing process. Treatments for flattening and softening scars include silicone gels and compression bandages. However, neither of these treatments can be applied to open wounds, and compression bandages require professional supervision (e.g., for large scars after skin grafts).

[0006] In summary, the current standard of care is to treat scar tissue that has already formed. Existing treatments are not suitable for treating open wounds or wounds that are in the early stages of the healing process with the goal of alleviating or reducing scar tissue formation in the first place. There are limited treatments available to reduce or reduce scar tissue formation in healing or early stages of the healing process (e.g., open wound treatment).

[0007] Sodium hypochlorite solutions have been used for wound irrigation to disinfect wounds. Examples of such solutions include Dakin's solution, which has been used as a disinfectant and typically contains 0.4% to 0.5% sodium hypochlorite and a buffer to adjust the pH to 9 to 10. Modified versions of Dakin's solution have been prepared as antibacterial solutions for wound disinfection.

[0008] Patent Documents 1 and 2 describe hypochlorite solutions used for the prevention or treatment of inflammatory reactions in uncontaminated or uninfected surgical sites, wounds, trauma sites, internal inflammatory lesions, or uninfected superficial inflammatory lesions. The hypochlorite solutions described therein may contain sodium hypochlorite in a concentration range of 0.005% to 0.2% by weight and may further contain sodium chloride in a concentration range of 0.5% to 1.5% by weight. Inflammatory conditions such as oral ulcers, eczema, and psoriasis are mentioned, and the treatment of inflammatory reactions in fistulas is exemplified. Neither Patent Document 1 nor Patent Document 2 contemplates the use of the hypochlorite solutions described therein for purposes other than reducing inflammatory reactions, nor for the prevention of scar tissue.

[0009] There is a need to provide new treatments to prevent or reduce the formation of scar tissue, and there is also a need for new treatments to improve the appearance of scar tissue. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] UK Patent Application Publication No. 2519774 [Patent Document 2] UK Patent Application Publication No. 2527264 Summary of the Invention

[0011] The inventors have surprisingly discovered that treating wounds with a 0.01% to 0.2% (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% [about 150 ppm to 600 ppm by weight]) hypochlorite solution is effective in the prophylactic treatment of scar tissue. Indeed, more specifically, the inventors have discovered that applying the hypochlorite solution described herein to new or recent wounds (i.e., wounds less than 10 days old) can dramatically reduce or even prevent the formation of scar tissue. The inventors have further discovered that application of the hypochlorite solution improves the overall appearance of the wound and minimizes the extent of scar tissue. Indeed, the inventors have surprisingly discovered that using the hypochlorite solution as described herein results in the formation of normal, healthy tissue in areas where scar tissue would normally be expected. Without wishing to be bound by theory, the inventors speculate that the formation of this normal healthy tissue is promoted by the unexpected and unpredictable effects of the hypochlorite solutions described herein on cytokines involved in scar tissue formation.

[0012] According to a first aspect of the present invention, there is provided a hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising a hypochlorite concentration ranging from about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% by weight [about 150 ppm to 600 ppm by weight]). As explained in more detail below, the inventors have surprisingly and unexpectedly discovered that the claimed hypochlorite concentrations modulate cytokines involved in scar tissue formation and promote cytokine activity associated with reduced scar formation. More specifically, the inventors have found that the hypochlorite solutions described herein promote interleukin (IL-10) activity, which is associated with reduced scar tissue formation. The inventors further demonstrate that the claimed solutions achieve reduced scar formation in clinical practice (see Examples).

[0013] The prophylactic treatment can include or consist of topical administration of a hypochlorite solution to a mammal, preferably a human, in need thereof. More specifically, the administration of the hypochlorite solution can include topical administration of the hypochlorite solution to a specific part of a mammal, preferably a human, in need thereof, where the specific part is a site of trauma or a site of a disease or condition characterized by abnormal collagen formation. For example, the prophylactic treatment can include topical administration of the hypochlorite solution to a wound or a site of trauma (e.g., a surgical trauma). For example, the prophylactic treatment can include topical administration of the hypochlorite solution to a part of a mammal, preferably a human, where the site is a site of a disease or condition characterized by abnormal collagen formation. The hypochlorite solution can be topically administered by irrigating, soaking, or washing the site for a certain period of time.

[0014] In embodiments, the prophylactic treatment can include administering a hypochlorite solution to a wound, and optionally the wound is a traumatic or surgical wound. Thus, the scar tissue to be prophylactically treated can be scar tissue resulting from wound healing, and optionally the wound is a traumatic or surgical wound. The wound may be sutured or may be an open wound. The hypochlorite solutions described herein are particularly effective for the prophylactic treatment of open wounds, for example, open wounds that have not yet closed through the natural healing process.

[0015] Preventive treatment can include administering hypochlorite solution to wound during the wound healing period.On the first day that hypochlorite solution is administered to wound, the wound is optionally less than 10 days old, optionally less than 5 days old, and even optionally less than 1 day old.For example, on the first day that hypochlorite solution is administered to wound, the wound can be less than 24 hours old, optionally less than 12 hours old, and optionally less than 6 hours old.Preferably, when the wound is fresh (i.e., within 1 hour, or within 2 hours, or within 3 hours after the wound is inflicted), the hypochlorite solution is administered to the wound immediately.Preventive treatment can include administering hypochlorite solution to wound by topical administration.

[0016] The scar tissue may be selected from one or more of keloid scar tissue, hypertrophic scar tissue, and scar contracture.

[0017] The scar tissue to be prophylactically treated may be scar tissue caused by a disease or condition characterized by abnormal collagen production. Diseases or conditions characterized by abnormal collagen production may be referred to as collagen diseases or collagen disease conditions. The collagen disease or collagen disease condition may be selected from a list including myocardial fibrosis and purpura fulminans. The collagen disease or collagen disease condition may be selected from pulmonary fibrosis, chronic obstructive pulmonary disease, and long-term COVID-19.

[0018] In embodiments, the mammal may be identified as being at risk for developing a lung disease or condition that causes scarring of the lungs, optionally the lung disease or condition is selected from pulmonary fibrosis, chronic obstructive pulmonary disease, and long-term COVID-19. For example, the mammal may be identified as being at risk for developing long-term COVID-19 after a positive diagnosis of COVID-19 infection. The prophylactic treatment may include administering a hypochlorite solution to the lungs by inhalation.

[0019] In embodiments, the collagen disease or collagen disease condition is not a respiratory condition or disease. In embodiments, the collagen disease or collagen disease condition is not a respiratory condition or disease selected from acute respiratory distress syndrome (ARDS); asthma; bronchitis; chronic obstructive pulmonary disease (COPD); the common cold; coronavirus diseases such as severe acute respiratory syndrome (SARS) and COVID-19; cystic fibrosis; influenza; Middle East respiratory syndrome (MERS); pneumonia, including viral pneumonia, bacterial pneumonia, and ventilator-associated pneumonia; pulmonary fibrosis; rhinovirus disease; sarcoidosis (e.g., affecting the lungs); tuberculosis; or inflammation of lung tissue.

[0020] Preventive treatment can include administering a hypochlorite solution to a mammal for several consecutive days. For example, the hypochlorite solution can be administered for at least two consecutive days, optionally at least three consecutive days, optionally at least four consecutive days, optionally at least five consecutive days, optionally at least six consecutive days, and optionally at least seven consecutive days. For example, the hypochlorite solution can be administered for at least 14 consecutive days, optionally at least 21 consecutive days, and optionally at least 30 consecutive days. Alternatively, the treatment can include administering a hypochlorite solution every other day for at least seven days, optionally at least 14 days, and optionally at least 28 days.

[0021] The prophylactic treatment can include administering the hypochlorite solution to the mammal at least once a day. The prophylactic treatment can include administering the hypochlorite solution to the mammal at least twice a day (e.g., twice a day or three times a day). The prophylactic treatment can include administering the hypochlorite solution to the mammal at least three times a day (e.g., three times a day or four times a day). The prophylactic treatment can include administering the hypochlorite solution to the mammal at least four times a day (e.g., four times a day or five times a day). The prophylactic treatment can include administering the hypochlorite solution to the mammal at least five times a day (e.g., five times a day or six times a day).

[0022] The hypochlorite solution can contain hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), where the hypochlorite is sodium hypochlorite.

[0023] The hypochlorite solution can contain hypochlorite at a concentration of about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), more preferably about 0.015% to 0.075% by weight (about 150 ppm to 750 ppm by weight), even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight), and most preferably about 0.04% to 0.06% by weight (about 400 ppm to 600 ppm by weight). The concentration of hypochlorite can be 0.045% to 0.055% by weight, for example, 0.05% by weight (500 ppm).

[0024] The hypochlorite solution can contain hypochlorite in a concentration range of about 0.0175% to 0.065% by weight (about 175 ppm to 650 ppm), for example, about 0.02% to 0.06% by weight (about 200 ppm to 600 ppm [e.g., about 200 ppm to 500 ppm]).

[0025] The hypochlorite solution can contain a hypochlorite concentration ranging from about 0.015% to 0.06% by weight (about 150 ppm to 600 ppm by weight), optionally about 0.02% to 0.055% by weight (about 200 ppm to 550 ppm by weight), further optionally about 0.02% to 0.05% by weight (about 200 ppm to 500 ppm by weight), further optionally about 0.025% to 0.05% by weight (about 250 ppm to 500 ppm by weight), and further optionally about 0.03% to 0.045% by weight (about 300 ppm to 450 ppm by weight). The hypochlorite concentration can be 0.035% to 0.04% by weight (about 350 ppm to 400 ppm by weight).

[0026] Without wishing to be bound by theory, it is believed that hypochlorite solutions with hypochlorite concentrations below 150 ppm, optionally 200 ppm, are less effective at reducing IL-6 function and maintaining IL-10 function than hypochlorite concentrations above 150 ppm, optionally 200 ppm. Hypochlorite concentrations above 150 ppm, optionally 200 ppm, have been shown to be particularly effective at reducing IL-6 function, compared to hypochlorite concentrations below 150 ppm (e.g., 100 ppm). Furthermore, without wishing to be bound by theory, it is believed that hypochlorite solutions with hypochlorite concentrations of 500 ppm or less do not adversely affect cells to which the hypochlorite solution is applied during use (e.g., no evidence of increased inflammation). Therefore, it is believed that hypochlorite solutions containing hypochlorite in a concentration range of 150 ppm to 500 ppm, optionally 200 ppm to 500 ppm, are particularly advantageous for use in embodiments of the present invention.

[0027] The hypochlorite solution further comprises sodium chloride, which may be present at a concentration ranging from about 0.2% to 3.0% by weight, optionally from about 0.3% to 1.5% by weight, and optionally from about 0.4% to 1.3% by weight. The present inventors have surprisingly discovered that providing a hypochlorite solution according to the present invention further comprising sodium chloride provides an unexpected improvement in reducing IL-6 function while maintaining IL-10 function, compared to a similar solution not containing sodium chloride (i.e., an aqueous solution containing hypochlorite).

[0028] While not wishing to be bound by theory, it is believed that providing a hypochlorite solution according to the present invention further comprising sodium chloride provides a stabilizing effect on the hypochlorite contained in the solution, preventing, for example, decomposition into other chlorine-containing species. Therefore, the inventors surprisingly discovered that by carefully controlling both the hypochlorite concentration and the sodium chloride concentration within the ranges disclosed herein (e.g., 150 ppm to 600 ppm hypochlorite and 0.2 wt. % to 3.0 wt. % NaCl, particularly 200 ppm to 500 ppm hypochlorite and 0.2 wt. % to 3.0 wt. % NaCl), the resulting hypochlorite solution is surprisingly stable and resistant to decomposition. Furthermore, advantageously, by carefully controlling both the hypochlorite concentration and the sodium chloride concentration, the need to provide a buffer in the hypochlorite solution can be eliminated. As described below, the hypochlorite solution according to the present invention does not need to be buffered and provides a beneficial regulatory effect on target cytokines involved in scar tissue formation.

[0029] The hypochlorite solution for use described herein can further comprise sodium chloride in a concentration range of about 0.5% to 1.2%, optionally in a concentration range of about 0.6% to 1.0% (e.g., 0.7% to 1.0%), and further optionally in a concentration range of about 0.8% to 1.0% by weight. The hypochlorite solution for use described herein may further comprise 0.8% by weight sodium chloride. The hypochlorite solution for use described herein may further comprise 0.85% by weight sodium chloride.

[0030] Hypochlorite solutions for use as described herein can further comprise sodium chloride in a concentration range of about 0.2% to 1.0%, optionally in a concentration range of about 0.3% to 0.8% (e.g., 0.3% to 0.7%), and further optionally in a concentration range of about 0.3% to 0.6% (e.g., 0.3% to 0.5%) by weight. Hypochlorite solutions for use as described herein can further comprise 0.4% by weight sodium chloride.

[0031] The hypochlorite solution may be an aqueous sodium hypochlorite solution containing sodium hypochlorite, sodium chloride, and the balance water.

[0032] Hypochlorite solution may be an aqueous sodium hypochlorite solution that is only composed of sodium hypochlorite, sodium chloride, and the remaining amount of water.Advantageously, without wishing to be bound by theory, it is believed that the hypochlorite solution that is only composed of the above-mentioned components means that other components do not inhibit the function of cytokines involved in wound healing process and scar tissue formation.In addition, without wishing to be bound by theory, it is believed that the presence of sodium chloride enhances the effect of the hypochlorite solution that regulates the function of cytokines involved in wound healing process and scar tissue formation compared with hypochlorite solution that does not contain sodium chloride.Therefore, only the components of the hypochlorite solution described herein affect the function of cytokines, thereby advantageously reducing the formation of scar tissue.

[0033] The hypochlorite solution can have a pH of about 5 to 11, preferably about 6 to 10, more preferably about 7 to 9, and even more preferably about 7 to 8. The hypochlorite solution may have a pH of about 7.5 to 11 (e.g., greater than 7.5 to 11), optionally about 8 to 10, further optionally about 8.5 to 9.5, and even further optionally about 8.5 to 9 (e.g., pH 9). Without wishing to be bound by theory, it is believed that providing a hypochlorite solution according to the present invention having a pH of 7.5 or greater provides unexpectedly improved efficacy in reducing IL-6 function while maintaining IL-10 function, compared to hypochlorite solutions having a lower pH.

[0034] The hypochlorite solution can include less than 0.005% (50 ppm) by weight hypochlorous acid, optionally less than 0.004% (40 ppm), further optionally less than 0.003% (30 ppm), and even optionally less than 0.002% (20 ppm). In such embodiments, the pH of the hypochlorite solution can be 7.5 or greater, e.g., 7.5-11, optionally about 8-10, and even optionally about 8.5-9.5 (e.g., about 8.5-9).

[0035] In the hypochlorite solutions described herein, the equilibrium is: [ka] The dynamic equilibrium between the hypochlorite ion ([OCl - The concentration of hypochlorous acid ([HOCl]) in the hypochlorite solution relative to the total concentration of hypochlorite ions ([OCl]) and hypochlorous acid ([HOCl]) can be less than 50% by weight, optionally less than 40% by weight, further optionally less than 30% by weight, and even more optionally less than 20% by weight at ambient temperature (i.e., about 20°C). Preferably, the concentration of hypochlorous acid ([OCl]) in the hypochlorite solution is less than 50% by weight, optionally less than 40% by weight, further optionally less than 30% by weight, and even more optionally less than 20% by weight. - The concentration of hypochlorous acid in the hypochlorite solution relative to the combined concentration of [HOCl] and hypochlorous acid ([HOCl]) is less than 10% by weight, more preferably less than 5% by weight, and even more preferably less than 1% by weight, at ambient temperature (i.e., about 20°C). In such embodiments, the pH of the hypochlorite solution may be 7.5 or greater, e.g., 7.5-11, optionally about 8-10, even optionally about 8.5-9.5 (e.g., about 9), even optionally about 8.5-9.

[0036] The hypochlorite solution can be substantially free of free chlorine (Cl). The hypochlorite solution can contain less than 0.0005% by weight (5 ppm) of free chlorine (Cl), optionally less than 0.0004% by weight (4 ppm), further optionally less than 0.0003% by weight (3 ppm), and even further optionally less than 0.0002% by weight (2 ppm) of free chlorine (e.g., less than 0.001% by weight [1 ppm]). In such embodiments, the pH of the hypochlorite solution can be 5.5 or greater, for example, between 5.5 and 11, optionally between about 7.5 and 11.

[0037] Hypochlorite solution can be unbuffered.That is, hypochlorite solution may not contain stabilizer, buffer, etc. Advantageously, without wishing to be bound by theory, it is believed that unbuffered hypochlorite solution means that stabilizer, buffer, etc. do not inhibit the function of cytokines involved in wound healing process.Therefore, only the components of hypochlorite solution described herein affect the function of cytokines, thereby advantageously reducing the formation of scar tissue.

[0038] The hypochlorite solution for use according to the present invention can be unbuffered and have a pH of about 5 to 11 (e.g., 7.5 to 11, or greater than 7.5 to 11), and can comprise (or consist of) 0.01% to 0.2% by weight sodium hypochlorite (e.g., 0.015% to 0.06%, e.g., 0.02% to 0.05% by weight), 0.2% to 3.0% by weight sodium chloride, and the balance water. The concentrations of sodium hypochlorite and sodium chloride can be as defined above according to the first aspect of the present invention. For example, the hypochlorite solution can be unbuffered and comprise (or consist of) 0.045% to 0.055% by weight sodium hypochlorite (e.g., 0.05% by weight), 0.8% to 1.0% by weight sodium chloride (e.g., 0.85% or 0.8% by weight), and the balance water. For example, the hypochlorite solution can be unbuffered and can comprise (or consist of) 0.045% to 0.055% (e.g., 0.05%) by weight sodium hypochlorite, 0.3% to 0.5% (e.g., 0.4%) by weight sodium chloride, and the balance water.

[0039] The hypochlorite solution for use according to the invention may be unbuffered, have a pH of about 9, and comprise (or consist of) 0.015% to 0.06% (150 ppm to 600 ppm) sodium hypochlorite by weight, 0.2% to 3.0% sodium chloride by weight, and the balance water. The concentrations of sodium hypochlorite and sodium chloride may be as defined above according to the first aspect of the invention.

[0040] For example, the hypochlorite solution can be unbuffered, can have a pH of about 9, and can comprise (or consist of) 0.02% to 0.05% by weight sodium hypochlorite, 0.8% to 1.0% by weight (e.g., 0.85% or 0.8%) sodium chloride, and the balance water.

[0041] For example, the hypochlorite solution can be unbuffered, have a pH of about 9, and contain (or consist of) 0.045% to 0.055% (e.g., 0.05%) sodium hypochlorite by weight, 0.8% to 1.0% (e.g., 0.85% or 0.8%) sodium chloride by weight, and the balance water. For example, the hypochlorite solution can be unbuffered, have a pH of about 9, and contain (or consist of) 0.045% to 0.055% (e.g., 0.05%) sodium hypochlorite by weight, 0.3% to 0.5% (e.g., 0.4%) sodium chloride by weight, and the balance water.

[0042] Accordingly, there is provided a hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising (or consisting of) 0.045% to 0.055% (e.g., 0.05%) by weight sodium hypochlorite, 0.8% to 1.0% (e.g., 0.85% or 0.8%) by weight sodium chloride, the balance water, optionally wherein treatment comprises topically administering the hypochlorite solution to an area of ​​the mammal in need thereof for at least 30 seconds at least once a day for at least 14 consecutive days.

[0043] Accordingly, there is provided a hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising (or consisting of) 0.045% to 0.055% by weight (e.g., 0.05%) sodium hypochlorite, 0.8% to 1.0% by weight (e.g., 0.85% or 0.8%) sodium chloride, and the balance water, optionally wherein treatment comprises administering the hypochlorite solution to a mammal in need thereof for at least 30 minutes at least twice a day, preferably at least three times a day, and more preferably at least four times a day, for at least two consecutive days.

[0044] Accordingly, there is provided a hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising (or consisting of) 0.045% to 0.055% by weight (e.g., 0.05%) sodium hypochlorite, 0.3% to 0.5% by weight (e.g., 0.4%) sodium chloride, and the balance water, optionally wherein treatment comprises topically administering the hypochlorite solution to an area of ​​the mammal in need thereof at least twice a day, preferably three times a day, for at least 5 minutes, preferably at least 30 minutes, for at least 21 consecutive days.

[0045] There is also provided a pharmaceutical composition for use in the prophylactic treatment of scar tissue in a mammal, more preferably a human, comprising a hypochlorite solution as defined according to the first aspect of the present invention.

[0046] There is also provided the use of a hypochlorite solution as defined in the first aspect of the invention in the preparation of a medicament for the prophylactic treatment of scar tissue in a mammal, preferably a human, wherein the treatment may be as defined by the first aspect of the invention.

[0047] Also provided is a method for the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising administering a therapeutically effective amount of a hypochlorite solution as defined by the first aspect of the present invention to a mammal in need thereof. The treatment may be as defined by the first aspect of the present invention.

[0048] There is also provided a method for improving the appearance of scar tissue, the method being cosmetic, comprising administering to a mammal an amount of a hypochlorite solution as defined according to the first aspect of the invention, wherein the administration is topical.

[0049] According to a second aspect of the present invention, there is provided a hypochlorite solution for use in the prevention or treatment of scar tissue in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, for example about 0.015% to 0.06% by weight [150 ppm to 600 ppm]). The hypochlorite solution may be as defined by the first aspect of the present invention. The prevention or treatment may be as defined by the first aspect of the present invention. For example, the prevention or treatment of scar tissue may be treatment of a wound. The prevention or treatment of scar tissue may be prophylactic treatment of a disease or condition characterised by abnormal collagen formation. The prevention or treatment of scar tissue may be treatment of purpura fulminans. The treatment may reduce scar tissue formation.

[0050] According to a third aspect of the present invention, there is provided a hypochlorite solution for use in treating purpura fulminans in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% by weight [150 ppm to 600 ppm]). The hypochlorite solution may be as defined by the first aspect of the present invention. The treatment may be as defined by the first aspect of the present invention.

[0051] According to a fourth aspect of the present invention, there is provided a hypochlorite solution for use in the prevention or treatment of a disease or condition characterized by abnormal collagen production in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% by weight [150 ppm to 600 ppm]). The hypochlorite solution may be as defined in the first aspect of the present invention. The treatment may be as defined in the first aspect of the present invention. Also provided is a method of preventing or treating a disease or condition characterized by abnormal collagen production (collagen disease), comprising administering a therapeutically effective amount of hypochlorite solution to a mammal, preferably a human, in need thereof, wherein the hypochlorite solution is according to the first aspect of the present invention. The treatment may be as defined in the first aspect of the present invention.

[0052] According to a fifth aspect of the present invention, there is provided a method for reducing collagen production in fibroblasts, the method comprising contacting the cells with a hypochlorite solution comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% by weight [150 ppm to 600 ppm]) for a time sufficient to reduce interleukin-6 (IL-6) activity and maintain interleukin-10 (IL-10) activity. In embodiments, the method is preferably not a treatment performed on the human or animal body (e.g., an in vitro or ex vivo method). Also provided is a method for preventing or treating a disease or condition that would benefit from reducing collagen production by fibroblasts, the method comprising administering a hypochlorite solution to a mammal, preferably a human, in need thereof, wherein the hypochlorite solution is according to the first aspect of the present invention. The treatment may be as defined by the first aspect of the present invention.

[0053] According to a sixth aspect of the present invention, there is provided a method of controlling fibroblast differentiation, the method comprising contacting cells with a hypochlorite solution comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight, e.g., about 0.015% to 0.06% by weight [150 ppm to 600 ppm]) for a time sufficient to reduce interleukin-6 (IL-6) activity and maintain interleukin-10 (IL-10) activity. In embodiments, the method is preferably not a treatment performed on the human or animal body (e.g., an in vitro or ex vivo method). Also provided is a method of preventing or treating a disease or condition that would benefit from controlling fibroblast differentiation, the method comprising administering a hypochlorite solution to a mammal, preferably a human, in need thereof, wherein the hypochlorite solution is according to the first aspect of the present invention. The treatment may be as defined by the first aspect of the present invention.

[0054] The invention is further illustrated in the following non-limiting drawings. [Brief explanation of the drawings]

[0055] [Figure 1] FIG. 1 shows IL-6 cells from Example 1 dialyzed against control saline (0.85% by weight) to determine cytokine functionality. [Figure 2] FIG. 1 shows IL-6 cells of Example 1 dialyzed against a solution of Composition 1 to determine the functionality of the cytokine. [Figure 3] FIG. 1 shows IL-6 cells from Example 1 dialyzed against a solution of Composition 2 to determine cytokine functionality. [Figure 4] FIG. 1 shows IL-10 cells from Example 2 dialyzed against control saline (0.85% by weight) to determine cytokine functionality. [Figure 5] FIG. 1 shows IL-10 cells of Example 2 dialyzed against a solution of Composition 1 to determine cytokine functionality. [Figure 6] FIG. 1 shows IL-10 cells of Example 2 dialyzed against a solution of Composition 2 to determine cytokine functionality. [Figure 7] FIG. 1 shows the viability of MC9 cells after exposure to IL-10 with A (composition 1) and B (composition 2). [Figure 8] FIG. 1 shows the viability of B9 cells after exposure to IL-6 in A (composition 1) and B (composition 2). [Figure 9] 10 is a photograph of the ankle of a patient according to Example 5. [Figure 10] 10A and 10B are photographs of the outer left thigh of a patient in Example 8, where FIG. 10A was taken before treatment, FIG. 10B was taken during treatment, and FIG. 10C was taken after 14 days of treatment. DETAILED DESCRIPTION OF THE INVENTION

[0056] The following embodiments apply to all aspects of the present invention.

[0057] The present invention will be further described below. In the following text, different aspects of the present invention are defined in more detail. Each aspect thus defined can be combined with any other aspect or aspects or embodiment or embodiments, unless expressly indicated to the contrary. In particular, any feature indicated as preferred or advantageous can be combined with any other feature or features indicated as preferred or advantageous.

[0058] The term "treatment," as used herein, unless otherwise indicated, means reversing, attenuating, alleviating, or inhibiting the progression of the disease or condition to which such term applies, or one or more symptoms of that disease or condition.

[0059] As used herein, the term "prophylactic treatment" refers to treatment designed to prevent the occurrence of a pathological condition or minimize the likelihood of the occurrence of a pathological condition. Prophylactic treatment of scar tissue includes preventing the formation of scar tissue as well as reducing the formation of scar tissue (e.g., reducing the surface area and / or severity).

[0060] The term "scar tissue" refers to tissue that forms during the wound healing process or as a result of a disease or condition that causes scarring of normal tissue. Scar tissue forms as a direct result of trauma to normal tissue, such as skin. Fibrosis is the process of forming new tissue in response to a wound, resulting in tissue remodeling and the formation of scar tissue. Thus, fibrosis is sometimes used in the art to mean the process by which scar tissue is formed. Fibrosis is promoted by fibroblasts. Therefore, when reference is made herein to the prophylactic treatment of scar tissue or the treatment of scar tissue, those skilled in the art will understand that the prevention or treatment of fibrosis is also contemplated.

[0061] The disease or condition characterized by abnormal collagen formation may be any disease or condition that causes scarring in tissue. Such diseases and conditions may be selected from myocardial fibrosis and purpura fulminans. Such diseases and conditions may include pulmonary fibrosis, chronic obstructive pulmonary disease, and long-term COVID-19.

[0062] The term "open wound" refers to an injury to the skin that has been punctured, lacerated, or cut, exposing internal body tissue. This term includes incisions, lacerations, abrasions, puncture wounds, and gunshot wounds. This term refers to a wound before it has closed through the natural healing process.

[0063] A "patient" includes humans, non-human mammals (eg, dogs, cats, rabbits, cows, horses, sheep, goats, pigs, deer, etc.), and non-mammals (eg, birds, etc.).

[0064] As used herein, "wt %" refers to mass weight relative to the total weight of the hypochlorite solution.

[0065] Without wishing to be bound by theory, it is believed that the hypochlorite solution of the present invention acts on known initiators, mediators, and regulators of scar tissue formation.The solution of the present invention is believed to inhibit the release of inflammatory substances (e.g., cytokines and chemokines) from platelets, but does not prevent platelet aggregation.It is also believed that this solution weakens the effects of cytokines, chemokines, and other inflammatory mediators.The proof-of-concept that such effects can be used for the preventive treatment of scar tissue has been demonstrated in the examples described herein.Furthermore, as demonstrated in the examples described herein, it has been found that such effects can be obtained without significant safety issues.

[0066] The present inventors have surprisingly and unexpectedly discovered that the hypochlorite solution described herein can selectively affect the function of certain anti-inflammatory cytokines.The data described herein demonstrate that the hypochlorite solution described herein abolishes the function of IL-6 in vitro.On the other hand, the hypochlorite solution described herein does not cause significant changes in the function of IL-10 in vitro.

[0067] Thus, the hypochlorite solution of the present invention can inhibit normal cytokine activity in vitro, thereby providing a means to regulate cytokine-dependent cell signaling pathways. IL-6 has been implicated in promoting scar tissue formation and fibrosis (Johnson et al.; DOI: 10.3390 / biomedicines8050101). IL-10 has been implicated in fibrotic cytokines, regulation of the extracellular matrix, and promotion of fibroblast function. IL-10 has therefore been shown to play a role in tissue remodeling and reduced scar formation (King et al.; DOI 10.1089 / wound.2013.0461). Without wishing to be bound by theory, these data suggest that use of the hypochlorite solution of the present invention maintains IL-10 function while losing IL-6 function, resulting in a net negative effect on scar tissue formation.

[0068] The present inventors have investigated the effects of hypochlorite ions (OCl - ) solutions have been found to be highly suitable at the concentrations described. Preferably, the hypochlorite solutions described herein contain chlorite ions (ClO - ) is not included.

[0069] The hypochlorite solution for use as described herein can be locally administered to a mammalian part in need thereof (for example, tissue at the site of trauma).The hypochlorite solution can be provided as a solution and can be locally applied by irrigating, soaking, or washing the tissue to be treated (for example, a wound) with the hypochlorite solution for a certain period of time.The hypochlorite solution can also be provided in the form of a cream or gel, or can be applied directly to the tissue to be treated (for example, a wound).

[0070] The hypochlorite solution can be topically administered for about 30 seconds to about 90 minutes. Preferably, the hypochlorite solution is topically administered for 1 minute to about 90 minutes, about 5 minutes to about 90 minutes, about 10 minutes to about 90 minutes, about 15 minutes to about 90 minutes, about 20 minutes to about 90 minutes, about 30 minutes to about 90 minutes, about 45 minutes to about 90 minutes, about 60 minutes to about 90 minutes, about 30 seconds to about 60 minutes, about 30 seconds to about 45 minutes, about 30 seconds to about 30 minutes, about 30 seconds to about 20 minutes, about 30 seconds to about 15 minutes, about 30 seconds to about 10 minutes, about 30 seconds to about 5 minutes, or about 30 seconds to about 1 minute.

[0071] The hypochlorite solution can be topically administered for at least 5 minutes, optionally at least 10 minutes, and even optionally at least 15 minutes. The tissue to be treated (e.g., the wound) can be irrigated, soaked, or washed with the hypochlorite solution for at least 20 minutes, and optionally at least 30 minutes.

[0072] The hypochlorite solution for use as described herein can be topically administered at least once a day.For example, prophylactic treatment can include topically administering the hypochlorite solution at least twice a day, optionally at least three times a day, and even optionally at least four times a day.

[0073] The hypochlorite solution can be topically administered for 2 to 30 consecutive days, optionally for 5 to 28 consecutive days, and further optionally for 7 to 14 consecutive days. Alternatively, treatment may involve topically administering the hypochlorite solution every other day for 2 to 30 consecutive days, optionally for 5 to 28 consecutive days, and further optionally for 7 to 14 consecutive days.

[0074] In embodiments, the hypochlorite solution is administered topically at least once per day for at least 5 consecutive days, optionally at least once per day for at least 7 consecutive days, and further optionally at least once per day (e.g., twice per day) for at least 14 consecutive days.

[0075] The hypochlorite solution for use described herein can be administered by inhalation, for example, via a nebulizer. The type of nebulizer is not particularly limited, as long as it can atomize the hypochlorite solution into aerosol droplets for inhalation. The nebulizer can be connected to an outlet configured to deliver the hypochlorite solution to a patient via the nose and / or mouth. For example, the outlet can be a mouthpiece or a face mask. The hypochlorite solution for use described herein can be administered by inhalation, for example, via an inhaler. For example, the inhaler can be a metered-dose inhaler. The metered-dose inhaler can be configured to deliver a set dose of hypochlorite solution with each actuation of the metered-dose inhaler. The hypochlorite solution for use described herein can be provided in a pharmaceutical container suitable for use with an inhalation device. The pharmaceutical container can include a cartridge or canister suitable for use with an inhalation device such as an inhaler or nebulizer. The hypochlorite solution for use described herein can be packaged in a cartridge containing a propellant, thereby forming a cartridge containing the hypochlorite solution and the propellant. Similarly, the hypochlorite solution can be packaged in a canister container containing a propellant, thereby forming a canister containing the hypochlorite solution and the propellant. The propellant can be a chlorofluorocarbon, a hydrofluoroalkane, or the like. The cartridge or canister can be used with an inhaler. The cartridge or canister may be removable from the inhaler. The cartridge or canister can be attached to the inhaler before use.

[0076] The hypochlorite solution can be administered continuously by inhalation. For example, such continuous administration by inhalation can be applied to patients on mechanical ventilation, hospitalized patients, sedated patients, or patients in a coma or induced coma. In some cases, the administration time can be from about 30 seconds to about 90 minutes. Preferably, the hypochlorite solution is administered by inhalation for about 1 minute to about 90 minutes, about 5 minutes to about 90 minutes, about 10 minutes to about 90 minutes, about 15 minutes to about 90 minutes, about 20 minutes to about 90 minutes, about 30 minutes to about 90 minutes, about 45 minutes to about 90 minutes, about 60 minutes to about 90 minutes, about 30 seconds to about 60 minutes, about 30 seconds to about 45 minutes, about 30 seconds to about 30 minutes, about 30 seconds to about 20 minutes, about 30 seconds to about 15 minutes, about 30 seconds to about 10 minutes, about 30 seconds to about 5 minutes, or about 30 seconds to about 1 minute. The hypochlorite solution can be administered by inhalation once, twice, three times, or four times per day. Preferably, the hypochlorite solution is administered once per day.

[0077] The hypochlorite solution can be administered in cycles including a treatment period in which the hypochlorite solution is administered once, twice, three or four times a day for a period of several days (e.g., about 1, 2, 3, 4, 5, 6 or 7 days) to several weeks (e.g., about 1, 2, 3 or 4 weeks), and optionally followed by a period in which the hypochlorite solution is not administered for a period of several days (e.g., about 1, 2, 3, 4, 5, 6 or 7 days) to several weeks (e.g., about 1, 2, 3 or 4 weeks). This cycle can be repeated at least twice. For example, this cycle can be repeated 2, 3, 4, 5, 6, 7, 8, 9, 10 times, etc.

[0078] The hypochlorite solution for use as described herein is preferably an aqueous solution.

[0079] Preferably, the hypochlorite solution contains hypochlorite at a concentration of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), more preferably about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), even more preferably about 0.015% to 0.075% by weight (about 150 ppm to 750 ppm by weight), even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight), and most preferably about 0.04% to 0.06% by weight (about 400 ppm to 600 ppm by weight). For example, the hypochlorite solution contains about 0.05% by weight (about 500 ppm by weight) of hypochlorite.

[0080] The hypochlorite solutions for use described herein can optionally further comprise sodium chloride at a concentration ranging from about 0.2% to 3.0% by weight, further optionally from about 0.3% to 1.5% by weight, and even further optionally from about 0.4% to 1.3% by weight. The hypochlorite solutions for use described herein can further comprise sodium chloride at a concentration ranging from about 0.5% to 1.2%, optionally from about 0.6% to 1.0% (e.g., 0.7% to 1.0%), and even further optionally from about 0.8% to 1.0% (e.g., 0.8% to 0.9%) by weight. The hypochlorite solutions for use described herein can further comprise 0.85% by weight of sodium chloride. Hypochlorite solutions for use as described herein may further comprise sodium chloride in a concentration range of about 0.2% to 1.0%, optionally in a concentration range of about 0.3% to 0.8% (e.g., 0.3% to 0.7%), and further optionally in a concentration range of about 0.3% to 0.6% (e.g., 0.3% to 0.5%) by weight. Hypochlorite solutions for use as described herein may further comprise 0.4% by weight sodium chloride.

[0081] The hypochlorite solution for use as described herein can have a pH of about 5 to 11, preferably about 6 to 10, more preferably about 7 to 9, and even more preferably about 7 to 8. In some embodiments, the pH may be about 5 to 6, about 6 to 7, about 8 to 9, about 9 to 10, or about 10 to 11. In embodiments, the pH of the hypochlorite solution is greater than 7.5, or the pH of the hypochlorite solution is 7.5 or greater. For example, in some embodiments, the pH of the hypochlorite solution is 10 to 11. The hypochlorite ion (ClO - An alkaline pH is usually preferred to ensure the presence of hypochlorite. When hypochlorite is acidified, a different chemical, hypochlorous acid, is produced.

[0082] Hypochlorite solutions for use as described herein can have a pH of about 7.5 to 11, optionally greater than 7.5 to about 11, further optionally about 8 to 10, further optionally about 8.5 to 9.5 (e.g., a pH of about 9), further optionally about 8.5 to 9.

[0083] The pH of hypochlorite solution is typically within the range encountered in normal physiological function and disease process when used.This is because hypochlorite solution preferably automatically adjusts pH.In other words, as will be described in more detail below, the hypochlorite solution described herein is preferably not buffered, so that when the hypochlorite solution described herein is applied to a patient who needs it (for example, to a part of the skin), the pH of the hypochlorite solution is adjusted to the pH of the part where it is applied.

[0084] The hypochlorite solution for use as described herein may not be buffered.In a preferred embodiment, the hypochlorite solution is not buffered.In other words, the hypochlorite solution does not contain a buffering agent.The hypochlorite solution preferably does not contain a stabilizer.Therefore, the hypochlorite solution does not contain other agents that may inhibit the beneficial effects described herein on the activity of IL-6 and IL-10.

[0085] The beneficial effects on the prevention or treatment of scar tissue are particularly pronounced when the patient is treated with an unbuffered solution comprising sodium hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), more preferably about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), and even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight), and sodium chloride in a concentration range of about 0.3% to 1.5% by weight, optionally about 0.4% to 1.3% by weight, further optionally about 0.5% to 1.2% by weight, and even more optionally about 0.8% to 1.0% by weight. Even more preferably, the solution comprises about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), more preferably about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), and even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight) of sodium hypochlorite, and sodium chloride in a concentration range of about 0.3% to 1.5% by weight, optionally in a concentration range of about 0.4% to 1.3% by weight, further optionally in a concentration range of about 0.5% to 1.2% by weight, and further optionally in a concentration range of about 0.8% to 1.0% by weight, and is unbuffered.

[0086] In particular, the beneficial effects on the prevention or treatment of scar tissue are particularly pronounced when the patient is treated with an unbuffered solution comprising sodium hypochlorite in a concentration range of about 0.015% to 0.06% by weight (about 150 ppm to 600 ppm by weight), more preferably about 0.02% to 0.5% by weight (about 200 ppm to 500 ppm by weight), and even more preferably about 0.025% to 0.045% by weight (about 250 ppm to 450 ppm by weight), and sodium chloride in a concentration range of about 0.3% to 1.5% by weight, optionally about 0.4% to 1.3% by weight, further optionally about 0.5% to 1.2% by weight, and still further optionally about 0.8% to 1.0% by weight, and having a pH of 7.5 or greater.

[0087] The beneficial effects on the prevention or treatment of scar tissue are particularly pronounced when the patient is treated with a solution comprising sodium hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), more preferably about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), and even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight), and sodium chloride in a concentration range of about 0.3% to 1.5% by weight, optionally about 0.4% to 1.3% by weight, further optionally about 0.5% to 1.2% by weight, and even more optionally about 0.8% to 1.0% by weight, buffered to a pH of about 5 to 11, preferably about 6 to 10.

[0088] For example, a hypochlorite solution comprising 0.85% sodium chloride and 0.05% (500 ppm) sodium hypochlorite (w / w) has been found to be highly beneficial in the prevention and treatment of scar tissue in patients, preferably mammals, and more preferably humans.

[0089] For example, a hypochlorite solution containing 0.4% sodium chloride and 0.05% (500 ppm) sodium hypochlorite (w / w) has been found to be highly beneficial in the prevention and treatment of scar tissue in patients, preferably mammals, more preferably humans.A hypochlorite solution containing 0.4% sodium chloride and 0.05% (500 ppm) sodium hypochlorite (w / w) has been found to be highly beneficial for use in the treatment of purpura fulminans.

[0090] Preferably, hypochlorite solution should be very pure.Preferably, hypochlorite solution is produced by electrolysis to ensure its purity and its safety and effectiveness.In an embodiment, hypochlorite solution comprises hypochlorite (for example, sodium hypochlorite), water, and optionally sodium chloride, with a purity of more than 90% by weight, optionally more than 95% by weight, further optionally more than 98% by weight, and further optionally more than 99% by weight based on the total weight of hypochlorite solution.

[0091] The hypochlorite solution can be prepared from a concentrated hypochlorite solution that is diluted before use. Preferably, the concentrated hypochlorite solution that is diluted before use is a concentrated sodium hypochlorite solution. The concentration of hypochlorite in the concentrated hypochlorite solution can be in the range of about 0.5% to 3% by weight. Furthermore, the concentrated hypochlorite solution can be buffered to a pH of about 9 to 15, preferably about 11 to 13. Alternatively, the concentrated hypochlorite solution can be unbuffered. The concentrated hypochlorite solution can be a stabilized sodium hypochlorite solution of 1% or 2% sodium hypochlorite, or, for example, a disinfectant solution known as "Milton's solution" containing sodium chloride. The diluted hypochlorite solution can be a 2.5% to 10% solution of Milton's solution diluted with water when the disinfectant solution is 2% sodium hypochlorite. The sodium chloride concentration in the solution is typically 16.5%. Therefore, the volume ratio of the hypochlorite solution to water can be in the range of 1:10 to 1:40. Alternatively, if the disinfectant solution is 1% sodium hypochlorite, the diluted hypochlorite solution can be a 5% to 20% solution of Milton's solution diluted with water. In this case, the volume ratio of the hypochlorite solution to water can be in the range of 1:5 to 1:20.

[0092] In either case, a predetermined amount of water and a predetermined amount of sodium hypochlorite solution can be used to obtain a stabilized sodium hypochlorite solution with a sodium hypochlorite concentration of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight), more preferably about 0.01% to 0.1% by weight (about 100 ppm to 1000 ppm by weight), and even more preferably about 0.025% to 0.075% by weight (about 250 ppm to 750 ppm by weight). The action of the sodium hypochlorite solution can stabilize the diluted disinfectant solution.

[0093] A suitable apparatus for preparing hypochlorite solutions for use in the present invention is described in WO 2011 / 128862.

[0094] Various embodiments and optional features have been described above, it will be understood that these embodiments and features can be combined in any feasible combination.

[0095] While the foregoing disclosure provides a general description of the subject matter encompassed within the scope of the present invention, including how to make and use the invention and its best mode, the following examples are provided to further enable those skilled in the art to practice the invention and to provide a more complete written description thereof. However, those skilled in the art will understand that the specifics of these examples should not be construed as limiting the invention, the scope of which should be understood from the claims appended to this disclosure and their equivalents. Various additional aspects and embodiments of the present invention will become apparent to those skilled in the art in light of the present disclosure. [Example]

[0096] Examples 1 and 2 - In vitro studies of cytokine function material: Solution of Composition 1—0.05% by weight sodium hypochlorite in 0.85% by weight saline Solution of Composition 2—0.1% by weight sodium hypochlorite in 0.8% by weight saline Solution of Composition 3—0.05% by weight sodium hypochlorite in 0.4% by weight saline IL-6-dependent cell line B9 (Cat. No. 12121201, murine B cell hybridoma) IL-10-dependent cell line MC9 (ARCC™ CRL-8306™, mouse liver mast cells) PrestoBlue™ (a cell-permeable fluorescent compound for use in cell viability assays)

[0097] Methods: Plasma samples were obtained from volunteers and cells were separated from the plasma. The plasma samples were placed in dialysis tubing with a pore size smaller than the molecular weight of the cytokine. This was then placed in a 1000-fold excess of dialysis medium (i.e., 1 ml in 1 L, 2 ml in 2 L, etc.).

[0098] The dialysis media was placed in either a control saline solution (0.85% by weight), Composition 1, or Composition 2, and extracted after 5, 15, 30, 45, and 60 minutes and then analyzed to determine cytokine functionality.

[0099] Cytokine Function: Functional cell assays were performed using patient plasma to examine the ability to support the survival of cytokine-dependent cell lines.

[0100] The serum was supplemented with human recombinant cytokines: heat-inactivated fetal bovine serum was used and human recombinant IL-6 and IL-10 were added at 100 ng / ml.

[0101] Serum samples were dialyzed against either the 0.85 wt % saline solution, Composition 1, or Composition 2 described above.

[0102] The function of IL-6 was assessed by determining its ability to sustain proliferation of the IL-6-dependent cell line B9.

[0103] The function of IL-10 was assessed by its ability to sustain proliferation of the IL-10-dependent cell line MC9.

[0104] Cell viability was measured using fluorescence. A cell-permeable, non-fluorescent compound was added to the assay. Live cells maintained a reducing environment and reduced the fluorescent compound, resulting in a color change that fluoresced at 560 nm.

[0105] The degree of fluorescence was measured, which directly correlates with the number of viable cells. The results are shown in Figures 1-8.

[0106] In some experiments, high levels of cytokines (i.e., 10 ng / ml) caused a decrease in cell viability. Without wishing to be bound by theory, the inventors speculate that this may be due to a prozone-like effect at high cytokine levels and / or that high cytokine levels may cause cell death or inhibition of cellular metabolism in this system.

[0107] Example 1 - IL6 1-3 show the results of cell viability assays of IL-6-supplemented serum samples dialyzed against a control saline solution (FIG. 1), composition 1 (FIG. 2), and composition 2 (FIG. 3).

[0108] In these graphs, the positive / negative boundary line represents the mean value of the negative control wells (no cytokine added, n=16) plus three standard deviations from this mean value, and any value above this value is considered positive.

[0109] Figures 2 and 3 show that dialysis of serum samples with Composition 1 or Composition 2 of the present invention results in loss of function of the IL-6 molecule, as measured by a cell viability assay.

[0110] Example 2 - IL10 4-6 show the results of cell viability assays of IL-10-spiked serum samples dialyzed against control saline solution (FIG. 4), composition 1 (FIG. 5), and composition 2 (FIG. 6).

[0111] In these graphs, the positive / negative boundary line represents the mean value of the negative control wells (no cytokine added, n=16) plus three standard deviations from this mean value, and any value above this value is considered positive.

[0112] This experimental data suggests that dialysis of serum samples with Composition 1 or Composition 2 results in a small loss of function of the IL-10 molecule as measured by cell viability assays, but that IL-10 remains functional. Thus, this loss of function is within the range predicted by the in vitro analysis, and IL-10 remains functional, as net survival is maintained.

[0113] To further evaluate this, the data was analyzed to examine the cell viability observed after exposure to Composition 1 or Composition 2 as a percentage of that achieved with the same amount of cytokine after dialysis of a 0.85 wt% saline solution (Figure 7), and the results were compared to those obtained for IL-6 analyzed in the same manner (Figure 8).

[0114] Figure 7 shows that IL-10 receptor binding is maintained, ensuring survival of MC9 cells. In contrast, Figure 8 shows that IL-6 function is almost completely lost after 15 minutes of exposure to Composition 1 or Composition 2. With the complete loss of IL-6 function and negligible impact on IL-10 function, the combined effect is significantly anti-inflammatory and anti-scar tissue formation. This includes the elimination of cytokine effects that promote scar tissue formation and the relative increase in cytokine effects that attenuate or reduce scar tissue formation. Thus, there is a net negative impact on scar tissue formation. In other words, there is a net anti-fibrotic effect.

[0115] Without wishing to be bound by theory, IL-6 may act as a pro-inflammatory and pro-fibrotic agent in fibrosis, countering the anti-inflammatory and anti-fibrotic effects of IL-10. Therefore, (at least partially) loss of IL-6 function attenuates IL-6's counteraction of IL-10 function and its associated downstream effects. Therefore, the ratio of IL-6 to IL-10, or increased relative levels of IL-10, can predict the likelihood of scar tissue formation. For example, King et al. (Advances in Wound Care, Vol. 3, No. 4, p. 315) reported that IL-10 is associated with wound healing and reduced scar formation. Additionally, Xi et al. (2017 PLoS 2017 12(6)) reported that IL-10 can be used as a marker to predict clinical outcomes in patients with sepsis-induced immunosuppression.

[0116] Without wishing to be bound by theory, it is believed that hypochlorite solutions according to the present invention, including Compositions 1 and 2, can help prevent scarring by promoting the function of IL-10 and decreasing the function of IL-6, thereby reducing or eliminating the effect of IL-6 in reducing the function of IL-10.

[0117] Example 3 - Facial Lacerations - In Vivo Patient Efficacy Study A woman in her 20s suffered multiple facial lacerations around her left eye and nose following a bicycle accident. Treatment included daily topical application of an aqueous solution of sodium hypochlorite in saline (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.85% by weight) for 14 consecutive days. Gauze pads were soaked in the sodium hypochlorite solution and applied to the lacerations. The facial lacerations healed completely within 4 weeks without evidence of scar tissue or skin discoloration.

[0118] Example 4 - Lip (cheek mucosa) laceration - in vivo patient efficacy study A woman in her 20s sustained lacerations on the inside of her upper and lower lips after a bicycle accident, which developed into stomatitis / mouth ulcers immediately after the accident. Treatment included daily topical application of sodium hypochlorite solution in saline (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.85% by weight) for 14 consecutive days. The sodium hypochlorite solution was used daily as a mouthwash. The mouth ulcers healed completely within 4 weeks without evidence of scar tissue or tissue discoloration.

[0119] Example 5 - Surgical Trauma - In Vivo Patient Efficacy Study Female, 60s. Surgical trauma following surgery on the fibular side of the left ankle to remove an internal fixation plate and screws. Treatment included immediate postoperative closure of the wound followed by daily topical application of a solution of sodium hypochlorite in saline (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.85% by weight) for 14 consecutive days. Gauze soaked in the solution was applied to the wound. The laceration healed completely within 4 weeks, leaving a very faint, thin scar (Figure 9).

[0120] Example 6 - Oral Surgical Trauma - In Vivo Patient Efficacy Study A man in his 20s suffered multiple oral trauma following dental surgery. Several teeth had been extracted, leaving exposed bone. Two weeks after surgery, the patient experienced poor healing and severe pain. He was enrolled in the study. Treatment included daily topical application of sodium hypochlorite solution in saline (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.85% by weight) for 14 consecutive days. The sodium hypochlorite solution was used daily as a mouthwash. The patient reported symptom relief and improved healing within one week of starting treatment, and further discontinued pain medication. The oral trauma sites completely healed within four weeks without evidence of scar tissue or tissue discoloration.

[0121] Example 7 - Long-term COVID 19 - In vivo patient efficacy study Treatment involved administering an aqueous solution of sodium hypochlorite in saline (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.85% by weight—referred to as Composition 3 in Example 7) delivered by a nebulizer.

[0122] Each enrolled patient had previously tested positive for COVID-19 and had been suffering from "long COVID-19" symptoms for at least one month from the date of their positive COVID-19 test. Each patient was administered Composition 3 via nebulizer for the duration shown in the table below. The frequency of inhalations is indicated as (number of times per day x duration of each treatment session). If administration was four times per day, Composition 3 was administered every six hours. If administration was three times per day, Composition 3 was administered every eight hours.

[0123] For patients 1 through 4, treatment was administered daily at the indicated frequency until the patient reported no symptoms. Treatment was administered for 21 consecutive days for patient 5 and for 7 consecutive days for patient 6 at the indicated frequency.

[0124] [Table 1]

[0125] Following treatment with Composition 3, patients 1 through 4, who had had long COVID-19 for six months or less, experienced relief from long COVID-19 symptoms. Two patients fully recovered from long COVID-19 (Patients 1 and 2), and one patient experienced a near-complete (90%) recovery (Patient 3), although mild asthma symptoms persisted.

[0126] In contrast, Patients 5 and 6, who had long-term COVID-19 for more than 12 months after treatment with Composition 3, experienced persistent long-term COVID-19 symptoms.

[0127] One patient (Patient 5), who had been off work due to the debilitating effects of long COVID-19, had had the disease for 24 months but partially recovered and experienced some symptomatic relief after treatment. Another patient (Patient 6), who had had long COVID-19 for 12 months, reported no substantial benefit after 7 days of treatment and confirmed lung scarring as a result of COVID-19 infection.

[0128] These data indicate that Composition 3, administered by inhalation, is an effective treatment for the symptoms of long-term COVID-19. While not wishing to be bound by theory, it is believed that patients who have long-term COVID-19 (e.g., longer than six months) suffer from severe lung damage and / or scarring. Accordingly, these data indicate that Composition 3 prevents severe lung scarring when long-term COVID-19 patients are treated early in the disease course (i.e., when patients are treated within six months of their positive COVID-19 test). In contrast, when severe scarring has already occurred (i.e., when patients are treated more than 12 months after their positive COVID-19 test), Composition 3 is less effective in treating long-term COVID-19, although some therapeutic benefit may still be achieved. Again, without wishing to be bound by theory, it is believed that this partial therapeutic benefit may be due to a reduction in scar tissue in the patient's lungs.

[0129] Example 8 - Purpura fulminans - In vivo patient efficacy study Female, under 10 years of age. The patient was hospitalized with chickenpox-associated purpura fulminans. This disease is a rare condition associated with high mortality, amputation, and tissue scarring. After hospitalization, the patient developed large purpuric skin lesions that could potentially become necrotic. Treatment involved immersing the patient in Composition 3, an aqueous sodium hypochlorite solution (NaOCl: 0.05% by weight (500 ppm), NaCl: 0.4% by weight), three times daily (every 8 hours), for 30 minutes each time. Treatment was administered for 21 consecutive days. The purpuric skin lesions completely healed without leaving any scarring. The patient made a complete recovery without any tissue scarring. Figures 10A-10C show images of a large purpuric skin lesion on the patient's left thigh. Figure 10A shows the lesion before treatment, Figure 10B shows the lesion during treatment, and Figure 10C shows the lesion 14 days after treatment. By day 21, the lesions had completely healed.

Claims

1. 1. A hypochlorite solution for use in the prophylactic treatment of scar tissue in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.015% to 0.06% by weight (about 150 ppm to 600 ppm by weight), having a pH of 7.5 or greater, and further comprising 0.2% to 3.0% by weight of sodium chloride.

2. 2. The hypochlorite solution for use according to claim 1, wherein the prophylactic treatment comprises topically administering the hypochlorite solution.

3. 3. The hypochlorite solution for use according to claim 1 or 2, wherein the prophylactic treatment comprises administering the hypochlorite solution to a wound, optionally wherein the wound is a trauma site or a surgical wound.

4. 4. The hypochlorite solution for use according to claim 3, wherein the wound is sutured or is an open wound.

5. 5. The hypochlorite solution for use according to claim 3 or 4, wherein the prophylactic treatment comprises administering the hypochlorite solution to the wound during the period in which the wound is healing.

6. 6. The hypochlorite solution for use according to any one of claims 3 to 5, wherein on the first day the hypochlorite solution is administered to the wound, the wound is less than 10 days old, optionally less than 5 days old, and further optionally less than 1 day old.

7. 7. The hypochlorite solution for use according to any one of claims 1 to 6, wherein the scar tissue is selected from one or more of keloid scar tissue, hypertrophic scar tissue, and scar contracture.

8. 2. The hypochlorite solution for use according to claim 1, wherein the prophylactic treatment is treatment of a disease or condition characterized by abnormal collagen production (collagen disease), and optionally the mammal has been identified as being at risk for developing a lung disease or condition that causes scarring of the lungs, and optionally the lung disease or condition is selected from pulmonary fibrosis, chronic obstructive pulmonary disease, and long-term COVID-19.

9. 9. The hypochlorite solution for use according to claim 8, wherein the prophylactic treatment comprises administering the hypochlorite solution to the lungs by inhalation.

10. 10. The hypochlorite solution for use according to any one of claims 1 to 9, wherein the prophylactic treatment comprises administering the hypochlorite solution to the mammal for several consecutive days.

11. 11. The hypochlorite solution for use according to any one of claims 1 to 10, wherein the prophylactic treatment comprises administering the hypochlorite solution to the mammal at least once a day, optionally at least twice a day.

12. The hypochlorite solution for use according to any one of claims 1 to 11, wherein the hypochlorite is sodium hypochlorite.

13. 13. The hypochlorite solution for use according to any one of claims 1 to 12, wherein the hypochlorite is in a concentration range of about 0.02% to 0.06% by weight, optionally about 0.025% to 0.06% by weight (about 250 ppm to 600 ppm by weight), further optionally about 0.04% to 0.05% by weight (about 400 ppm to 500 ppm by weight).

14. 14. The hypochlorite solution for use according to any one of claims 1 to 13, wherein the sodium chloride is in a concentration range of 0.3wt% to 1.5wt%, optionally in a concentration range of about 0.4wt% to 1.3wt%.

15. 15. The hypochlorite solution for use according to any one of claims 1 to 14, which is an aqueous sodium hypochlorite solution comprising sodium hypochlorite, sodium chloride, and the balance water, optionally consisting only of sodium hypochlorite, sodium chloride, and the balance water.

16. 16. The hypochlorite solution for use according to any one of claims 1 to 15, having a pH of about 7.5 to 11 (e.g., greater than 7.5 up to 11), optionally about 8 to 10, and further optionally about 8.5 to 9.

5.

17. A hypochlorite solution for use according to any one of claims 1 to 16, which is unbuffered.

18. 18. A pharmaceutical composition for use in the prophylactic treatment of scar tissue in a mammal, more preferably a human, comprising a hypochlorite solution according to any one of claims 1 to 17.

19. 19. A method for improving the appearance of scar tissue, the method comprising administering to a mammal, preferably a human, an amount of the hypochlorite solution of any one of claims 1 to 17 or the pharmaceutical composition of claim 18, the method being a cosmetic method, wherein the hypochlorite solution is administered topically.

20. 18. A hypochlorite solution for use in the prevention or treatment of scar tissue in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.015% to 0.06% by weight (about 150 ppm to 600 ppm by weight), having a pH of 7.5 or greater, and further comprising 0.2% to 3.0% by weight of sodium chloride, optionally according to any one of claims 1 to 17.

21. 21. The hypochlorite solution for use according to claim 20, wherein the prevention or treatment of scar tissue is the treatment of a wound.

22. 21. The hypochlorite solution for use according to claim 20, wherein the prevention or treatment of scar tissue is prophylactic treatment of a lung disease or condition that causes scarring of the lungs.

23. 21. The hypochlorite solution for use according to claim 20, wherein the prevention or treatment of scar tissue is the treatment of purpura fulminans.

24. 1. A hypochlorite solution for use in the treatment of purpura fulminans in a mammal, preferably a human, comprising hypochlorite in a concentration range of about 0.01% to 0.2% by weight (about 100 ppm to 2000 ppm by weight).

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