NZ721272B2 - Composition, system and method for treating skin - Google Patents

Abstract

The cosmetic and non-cosmetic uses of a topical dry powder composition for wound healing and/or skin treatment comprises honey, a bicarbonate salt and an additional organic acid all in a dry granular form. The salt is capable of releasing CO2. The composition may optionally comprise thickeners.

Description

COMPOSITION, SYSTEM AND METHOD FOR TREATING SKIN 2 CROSS REFERENCE TO PRIOR APPLICATIONS 3 [0001] The present invention claims priority to US Application Number 61/908,611, filed 4 on November 25, 2014. NZ 759509 was divided out of the present application and the entire contents of each application are incorporated herein by reference. 6 FIELD OF THE INVENTION 7 [0002] The present invention relates to a topical compositions, systems and methods for 8 treating skin. In particular, the invention relates to the treatment of wounds and skin 9 disorders and to cosmetic skin treatment. More particularly, the invention relates to topical compositions and use of such compositions for promotion of wound healing, and removal of 11 non-viable body tissue, including removal of necrotic tissue from wound sites, and superficial 12 exfoliation of the epidermis for cosmetic improvement of skin appearance. 13 BACKGROUND OF THE INVENTION 14 [0003] Removal of non-viable body tissue is necessary or desirable for a number of reasons. Necrotic or non-viable tissue can develop within wound sites, including ulcerations 16 in skin tissue, which may be due to chronic pressure and vascular insufficiency, and wounds 17 due to physical trauma, burns (chemical or thermal), and sites of infection. Necrotic tissue 18 inhibits the normal progression of healing by physically and biochemically obstructing the 19 cascade of healing events in a wound site, and commonly harbors pathogens and their toxins. Removal of necrotic or non-viable skin tissue is a key step in healing such wounds. 21 When healing medical devices such as Negative Pressure Wound Treatment (“NPWT”) are 22 indicated, it is important to thoroughly debride the wound of necrotic tissue and fibrotic tissue 23 prior to using NPWT. This process is referred to as debridement. Debriding of the necrotic 24 tissue is a first priority in healing the wound and is commonly accomplished by one or more of the following methods: (1) surgical debridement; (2) mechanical debridement; (3) 26 enzymatic debridement; and (4) autolytic debridement. Surgical debridement and 27 mechanical debridement, while effective, are often somewhat non-specific and can result in 28 excessive tissue removal and pain to the patient. 29 [0004] Autolytic debridement involves the use of the body’s own proteolytic enzymes (proteases) and moisture to rehydrate and digest necrotic tissue. It is typically only used in 31 cases where the amount of necrotic tissue is relatively small and the slow rate of self- 32 debridement is acceptable.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [0005] Enzymatic debridement, using proteolytic enzymes such as pepsin or 2 papain/urea, is highly selective for non-viable tissue only. Examples of known enzymatic 3 debridement methods and compositions include those taught in U.S. Pat. No. 6,548,556 and 4 US Patent Application Publication No. 2009/0010910, as well as commercially known compositions such as Accuzyme®. Although enzymatic debridement compositions and 6 methods are known, there are a number of drawbacks and disadvantages to these 7 compositions. 8 [0006] Adverse or harmful reactions have been reported in patients using topical 9 enzymatic debriding compositions comprising papain as the active ingredient, including hypersensitivity reactions leading to hypotension (low blood pressure) and tachycardia (rapid 11 heart rate). There is also a problem with possible allergic reactions. Papain is derived from 12 papaya, and debriding compositions based on papain may also contain residual extracts 13 from papaya. In addition, patients who are allergic to latex may exhibit cross-reactivity with 14 papaya extracts. An alternative to papain is pepsin, a digestive enzyme. However, pepsin is most active in acidic environments (e.g. pH 2-3) such as the stomach, and is relatively 16 inactive at pH 6.5 and higher. As a result, pepsin has very limited activity in the more neutral 17 pH found in other body tissues such as skin and it is not very effective in debriding wound 18 sites. In addition, commercially available pepsin is typically obtained from pig stomach, and 19 is therefore not acceptable to some patients.

[0007] Another issue with topically applied enzymatic compositions is the ability of the 21 composition to effectively coat and penetrate the wound site, and deliver the enzyme to the 22 entirety of the wound site. In practice, this can be difficult, particularly if the wound site 23 penetrates deeply into the body or encompasses multiple layers of different body tissues. 24 [0008] Removal of dead skin cells from the outermost layer (the stratum corneum) of the epidermis is desirable in certain cosmetic applications. Skin tissue is composed of two 26 layers, the dermis and the epidermis. The dermis contains hair follicles, nerve endings, 27 blood vessels, and oil and sweat glands. The epidermis is the outermost layer and unlike 28 the dermis, it does not contain blood vessels. The epidermis is composed of a number of 29 different layers and within the innermost (basal) layers of the epidermis are actively dividing skin cells known as keratinocytes. With each round of cell division, a generation of cells is 31 pushed to the next level upwards, until they finally reach the outermost layer of the 32 epidermis, the stratum corneum. The stratum corneum is composed of a thin layer of 33 flattened, dead skin cells filled with keratin, and bound together by lipids. The outermost PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 layer of dead skin cells of the stratum corneum sheds off naturally, and is continually 2 replaced by newer layers below. On average, it takes about 28 days for a newly divided 3 daughter cell to migrate from the basal layer of the epidermis to the outermost layer of the 4 stratum corneum, where it can be shed off; this process is known as cell turnover. However, the rate of cell turnover can be affected by a number of factors, including hormonal 6 fluctuations, increasing age, poor health and the local environment (e.g. dry weather). A 7 build-up of dead skin cells in the outermost layer of skin can cause the skin to appear dull, 8 dry and ashy. In addition, a build-up of dead skin cells and sebum may plug pores, providing 9 an ideal site for bacterial infection, which then leads to acne breakouts. Superficial exfoliation of the outermost layer of skin to remove dead skin cells is thus highly useful for 11 cosmetically improving skin appearance, and it is also useful for overcoming active acne and 12 preventing future acne breakouts. 13 [0009] Exfoliation of skin tissue can be carried out by mechanical means or by chemical 14 means. Examples of mechanical methods of exfoliation include hand-held tools such as scrubbing brushes, cloths and sponges, as well as cleansing compositions containing small 16 beads or granules that act as a gentle abrasive when applied to the skin. However, there 17 are risks associated with mechanical methods of exfoliation: users may scrub too hard or too 18 often, and cause damage to the skin. This may result in irritation or drying out of the skin, 19 which can lead to further problems with the skin.

[0010] The most common examples of chemical-based methods of cosmetic exfoliation 21 include organic acids and retinoids. Weak organic acids, including beta hydroxy acids (e.g. 22 salicylic acid) and alpha hydroxy acids, work as keratolytic agents, loosening and softening 23 the outermost layer of the epidermis and thus helping to slough off dead skin cells. Alpha 24 hydroxy acids, sometimes referred to as fruit acids, include glycolic acid, lactic acid, citric acid, mandelic acid and malic acid. Low concentrations of alpha and/or beta hydroxy acids 26 are available in a variety of over-the-counter (OTC) cosmetic products, such as masks, 27 creams, cleansers, lotions and toners. Application of such acids can cause stinging, 28 redness and irritation, particularly those individuals with sensitive skin. Stronger organic 29 acids, such as phenol and trichloroacetic acid, and higher concentrations of glycolic acid (e.g. >10%), penetrate the skin more deeply and can cause chemical burns. As such, these 31 acids are not available for over-the-counter use and may only be applied by experienced 32 medical professionals. Retinoids are a class of chemical compounds that trigger an increase 33 in the rate of cell turnover. The use of retinoids is not without risk: there are numerous side 34 effects associated with retinoid use, include sun sensitivity, redness, irritation, and peeling, PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 flaking skin. A number of retinoid compounds are only available through prescription by a 2 doctor. As well, retinoids, particularly oral forms, are contraindicated during suspected or 3 confirmed pregnancy, due to the known association of retinoids with defects in fetal 4 development. In addition to the above, benzoyl peroxide is commonly used for acne treatment, as it is both antimicrobial and an exfoliant. However, it often causes skin irritation 6 and dryness. 7 [0011] Exfoliation of the epidermis with concomitant moisturization is necessary to 8 manage certain chronic skin conditions such as ichthyosis, psoriasis, seborrheic dermatitis 9 and eczema. Current treatments for these skin conditions typically include corticosteroids, coal tar, occlusive ointments and creams. In certain cases of icthyosis and psoriasis, 11 retinoids may be prescribed. However, coal tar is often a treatment of last resort, due to the 12 unpleasant smell and the unknown risks of long-term, low level exposure to potential 13 carcinogens found in coal tar. Corticosteroids cannot be used for long periods of time, as it 14 can cause permanent thinning of the skin and telangiectasia.

[0012] Honey has historically been used to treat a wide variety of medical concerns, 16 including wounds, gastrointestinal disorders, allergies and infections of the upper respiratory 17 tract. It is known to promote the healing of wounds as it provides numerous beneficial 18 effects, including being anti-inflammatory, antioxidant, antimicrobial and anti-bacterial. 19 [0013] Honey is composed primarily of sugars, mainly fructose and glucose. Most types of honey are acidic due to the presence of naturally occurring organic acids, and have an 21 average pH of around 3.9, but can range anywhere from pH 3.2 to 6.1. The water content in 22 honey is very low, with an average water activity (aw) of 0.6; the water that is present within 23 honey is bound to the sugar molecules that make up the bulk of honey, thus making honey 24 inhospitable to microbial growth. The antimicrobial effect of honey is mainly attributed to the following factors: its low water content, hydrogen peroxide (H2O2), methylglyoxal, the 26 antimicrobial peptide bee defensin-1, and its acidic pH. 27 [0014] Honey facilitates wound healing through its strong osmotic action on body tissue: 28 as honey contains very little moisture (i.e. has a low water activity), it draws wound exudate 29 by osmosis from the surrounding tissue. By drawing exudate out of the wound site, there is a constant supply of proteases brought to the wound site, particularly at the interface 31 between necrotic tissue and viable tissue. As a result, autolytic debridement or desloughing 32 of non-viable body tissue can be significantly enhanced with the use of honey as a wound PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 dressing. In addition, the absorption of wound exudate by honey inhibits colonization of the 2 wound site with microbial growth. 3 [0015] Honey also enhances wound healing by inhibiting microbial growth within the 4 wound site, and inhibiting oxidative reactions that cause oxidative stress and inflammation.

In most types of honey, the acidic pH, low water content and enzymatic production of 6 hydrogen peroxide are the main factors contributing to the antimicrobial activity. However, 7 for any given type of honey, the factors contributing to the antimicrobial activity and the 8 overall level of antimicrobial activity are variable and highly dependent on the plant species 9 that the honey bees originally fed upon. Honey derived from bees that have fed exclusively on Leptospermum scoparium, commonly referred to as Manuka honey, as well as honey 11 derived from bees that have fed on other species of Leptospermum species, is known to 12 have exceptional anti-microbial effect [1]. Manuka honey, as produced from Leptospermum 13 species and particularly Leptospermum scoparium, contains significantly higher 14 concentrations of methylglyoxal compared to other types of honey. The high concentration of methylglyoxal in Manuka honey is believed to be a major contributing factor to the 16 observed antimicrobial effect [2]. Concentrations of methylglyoxal in Manuka honey can 17 vary widely, dependent on the bees and the cultivar(s) of Leptospermum species that have 18 been fed upon; however, a minimum methylglyoxal concentration of around 150 mg/kg is 19 believed to be directly responsible for the observed antimicrobial effect [3]. The antimicrobial effect due to methylglyoxal in Manuka honey is a non-peroxide effect that is sometimes also 21 referred to as the “Unique Manuka Factor” (“UMF”), a commercial rating scale that compares 22 observed antimicrobial activity to solutions of phenol in water (e.g. a UMF of 10 is equivalent 23 to the antimicrobial effect of 10% v/v phenol in water). 24 [0016] Although honey has numerous beneficial effects on wound treatment, it is typically in the form of a flowable liquid and can be difficult to keep in place, particularly when 26 treating chronic wounds that may require long-term care. Specialized dressings may be 27 required. For example, U.S. Patent No. 7,714,183 discloses a wound dressing impregnated 28 with honey, as a means of keeping honey secure over the wound site. However, such 29 dressings may actually inhibit wound healing by slowing down collagen deposition [4]. Also, as honey is typically acidic, the application of honey to a wound site may cause pain or 31 discomfort to the patient. 32 [0017] There is evidence that carbon dioxide gas is beneficial for treating wound sites 33 and promoting wound healing [5]. This is believed to be due to the Bohr effect, wherein PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 hemoglobin’s binding affinity to oxygen is inversely related to local acidity and carbon dioxide 2 concentration. With an increase in local carbon dioxide concentration, the binding affinity of 3 hemoglobin decreases. In addition, the body reacts by increasing blood flow to areas where 4 local carbon dioxide concentration is increased. This may be observed as an increase in microcirculation of blood flow surrounding the wound area: this increase in circulation 6 provides greater local oxygen availability and nutrient flow to the wound site, while also 7 helping to removing wastes from the wound site. A carbon dioxide-enriched environment 8 also minimizes the possibility of infection by aerobic bacteria. At the same time, there is no 9 toxicity or major side effects to the patient to applying carbon dioxide. However, a disadvantage of carbon dioxide therapy is the need for specialized equipment in order to 11 create a localized environment surrounding the wound site that can be filled with a sufficient 12 concentration of carbon dioxide to observe an effect. This requires a source of carbon 13 dioxide (usually in the form of a pressurized gas cylinder) and a method of delivering carbon 14 dioxide to form a localized environment around the wound site, which may in turn require tubing and a syringe to inject carbon dioxide into the wound site and a physical method to 16 enclose the wound site, e.g. adhesive dressings. As a result, carbon dioxide therapy is not a 17 method that can be readily carried out without specialized equipment, and often requires the 18 assistance of a skilled practitioner. An alternative, however, is provided in US 19 2006/0150988, which describes a cosmetic treatment method wherein carbon dioxide is transdermally delivered using a polymer solution. 21 [0018] There exists a need for methods and agents for removing non-viable or necrotic 22 tissue from wound sites that overcomes at least one of the deficiencies known in the art. 23 There also exists a need for compositions for exfoliation of the skin and/or to improve skin 24 appearance, which overcomes at least one of the deficiencies known in the art.

SUMMARY OF THE INVENTION 26 [0019] In one aspect, the present invention provides compositions that may be used as 27 desloughing agents, debriding agents, wound treatment agents, veterinary wound care 28 agents, chronic skin disorder treatment agents, cosmetic skin exfoliants or any combination 29 thereof.

[0020] In one aspect, the invention provides a topical composition comprising: 31 [0021] - honey; and, 1 [0022] - a pharmaceutically acceptable carbonate (CO3-2) and/or bicarbonate 2 (HCO3-1) salt; 3 [0023] for use as a desloughing agent, a debriding agent, a wound treatment agent, 4 a veterinary wound care agent, a chronic skin disorder treatment agent, a cosmetic skin exfoliant or any combination thereof. 6 [0024] In accordance with an aspect of the present invention, there is provided a 7 composition comprising: 8 [0025] - honey; 9 [0026] - a pharmaceutically acceptable salt of carbonate (CO ) or bicarbonate (HCO ), that is capable of reacting with an acid or an acidic solution to form carbon dioxide; 11 and 12 [0027] wherein the pH of said composition is about 7.0 to 7.4. 13 [0028] The compositions described herein may be used as a desloughing agent, a 14 debriding agent, a wound treatment agent, a veterinary wound care agent, a chronic skin disorder treatment agent, and a cosmetic skin exfoliant. 16 [0029] In one embodiment of the present invention, the composition comprises 17 [0030] - honey; 18 [0031] - about 2.5 to 20% w/w, preferably about 7.5 to 12.5% w/w, sodium 19 bicarbonate;

[0032] - about 0.2 to 6% w/w, preferably about 2 to 5% w/w, xanthan gum; and 21 [0033] - about 0.2 to 10% w/w, preferably about 1 to 5% w/w, ascorbic acid. 22 [0033A] In a particular aspect, the present invention provides use of a dry powder 23 composition in the manufacture of a medicament for desloughing, debriding, wound treating, 24 or exfoliating applications, the composition comprising: [0033B] - honey, wherein the honey is in a dry granular form; 26 [0033C] - at least one pharmaceutically acceptable bicarbonate (HCO3 ) salt in 27 a dry granular form; and (followed by page 7a) [0033D] - at least one organic acid in dry granular form, wherein the at least one organic acid is additional to organic acids naturally occurring in honey; [0033E] wherein the medicament is formulated for topical administration, and provided that the use is not cosmetic [0033F] In another particular aspect, the present invention provides a cosmetic method of desloughing, debriding or exfoliating a region of skin of a mammal, the method comprising applying to the region a topical composition comprising: [0033G] - honey in dry granular form; [0033H] - at least one pharmaceutically acceptable bicarbonate (HCO ) salt in dry granular form; and, [0033I] - at least one organic acid in dry granular form, wherein the at least one organic acid is additional to organic acids naturally occurring in honey.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention will be further understood from the following detailed description of an embodiment of the invention, with reference to the drawings in which: Figure 1 is a graph of observed desloughing in vitro of a sample of pig skin over time, with compositions of formulas (A) to (E) as denoted in Example 1; the concentrations of [FOLLOWED BY PAGE 8] PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 sodium bicarbonate in honey by percentage weight in each of formulas (A) to (E) are 2 denoted as “HoneyMousse [concentration of NaHCO , % w/w]” in the legend; 3 [0036] Figure 2 is a graph of observed desloughing in vitro of a sample of pig skin over 4 treatment time, comparing the desloughing efficiency of Formulation (E) (as described in Example 1), compared with honey alone (control), papain ointment and Santyl collagenase 6 ointment; 7 [0037] Figure 3 is a series of photographs of observed changes in vivo, in a myiasis 8 ulcer on a leg of a donkey, taken at Day 0, Day 7, Day 29 and Day 63, counting from the first 9 day of treatment as described in Example 1(ii)(a);

[0038] Figure 4 is a series of photographs of observed changes in vivo, in an ankle 11 wound on a donkey, taken at Day 0, Day 3 and Day 18, counting from the first day of 12 treatment as described in Example 1(ii)(b); 13 [0039] Figure 5 is a series of photographs of observed changes in vivo, in a myiasis 14 ulcer on the girth of a horse, taken at Day 0, Day 16, Day 21 and Day 32, counting from the first day of treatment as described in Example 1(ii)(d); and 16 [0040] Figure 6 is a series of photographs of observed changes in vivo, in a surgical 17 excision wound of a human male patient (following excision of a keratoacanthoma), taken at 18 Day 0, Day 6 and Day 7, counting from the first day of treatment as described in Example 19 1(ii)(c).

DETAILED DESCRIPTION OF EMBODIMENTS 21 [0041] The terms "comprise", "comprises", "comprised" or "comprising" may be used in 22 the present description. As used herein (including the specification and/or the claims), these 23 terms are to be interpreted as specifying the presence of the stated features, integers, steps 24 or components, but not as precluding the presence of one or more other features, integers, steps, components or a group thereof as would be apparent to persons having ordinary skill 26 in the relevant art. 27 [0042] The term “about” as used herein with respect to concentrations, pH or other 28 quantities is intended to indicate a variation of at least 10%. 29 [0043] As used herein (including the specification and/or the claims), the terms “debriding” and “debridement” refer to the removal of damaged, non-viable or necrotic tissue PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 from a wound, and the term “slough” refers to the shedding of non-viable or necrotic tissue 2 from living tissue. As used in herein, the terms “desloughing agent” and “debriding agent” 3 refer to agents and compositions that remove, or aid in the removal of, non-viable or necrotic 4 tissue from living tissue, by facilitating detachment of slough from living tissue within a wound site. 6 [0044] The present invention is based on the finding that a composition comprising 7 honey and one or more pharmaceutically acceptable mineral salts of carbonate (CO ) or 8 bicarbonate (HCO ) (the salts capable of reacting with (1) moisture and an acid, or (2) an 9 acidic solution, to form carbon dioxide), provides an unexpected significant enhancement of the removal of non-viable tissue, that is far greater than observed for either honey alone or 11 carbon dioxide alone. The removal of non-viable tissue may be for the purposes as a 12 desloughing, wherein the composition is used as a debriding agent, for the purposes of 13 promoting wound healing, or it may be for superficial exfoliation of the epidermis to 14 cosmetically enhance skin appearance. The enhancement of wound healing and therapeutic effect has been found to be far greater than would be expected from a simple 16 additive effect of honey and carbon dioxide used separately to treat a wound site. 17 [0045] Honey contains naturally occurring organic acids, including gluconic acid, lactic 18 acid, acetic acid, butyric acid, citric acid, malic acid, pyroglutamic acid and succinic acid. 19 Gluconic acid is present in the highest concentration (up to 1% w/w), as it is produced by the oxidation of glucose by glucose oxidase, both of which are naturally occurring in honey. 21 Medical grade Manuka honey typically has a pH in the range of pH 3.2 to 4.5. The 22 concentration of total organic acids in Manuka honey from Leptospermum scoparium ranges 23 from 0.17% w/w to 1.17% w/w. If desired, the pH of the honey may be further lowered by 24 addition of any of the above-noted acids that naturally occur in honey, as well as other food grade or pharmaceutically acceptable weak organic acids such as ascorbic acid, and the 26 alpha hydroxy acids, glycolic acid, citric acid and mandelic acid. In a preferred embodiment, 27 gluconic acid and/or lactic acid, or salts thereof, may be added to the composition of the 28 invention, as both of these acids and their salts provide beneficial effects. In the case of 29 gluconic acid, it is capable of chelating metal ions, which may help to inhibit oxidative reactions that may otherwise damage healing tissue. In addition, both gluconic and lactic 31 acids are humectants and thus provide moisturizing properties. In another preferred 32 embodiment, the composition may further comprise ascorbic acid or a salt thereof. Ascorbic 33 acid and its salts act as antioxidants and thus limit oxidate stress within the wound site, 34 thereby promoting wound healing.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [0046] In a preferred embodiment, the honey used to prepare the composition is medical 2 grade Manuka honey, with a minimum UMF rating of 10 (equivalent to antimicrobial effect of 3 10% phenol in water), a pH in the range of about 3.2 to 4.5, and a minimum methylglyoxal 4 concentration of about 150 mg/kg. Preferably, the total organic acid content is in the range of about 0.17-1.17% w/w, with gluconic acid being the primary organic acid. 6 [0047] The addition of a pharmaceutically acceptable mineral salt of carbonate (CO ) or 7 bicarbonate (HCO ) to honey, initiates a reaction between the salt and the naturally 8 occurring moisture and acids within the honey that evolves carbon dioxide gas within the 9 honey. In a preferred embodiment, the composition according to one embodiment of the invention is in the form of a foam or mousse, wherein the foam comprises fine bubbles 11 enriched in carbon dioxide. A slight molar excess of the mineral salt to the naturally 12 occurring acids within the honey is added, such that the resultant composition has a pH that 13 is preferably in the range of about 6.3-7.4. 14 [0048] The above-noted composition may be used as a desloughing composition, or debriding composition. The composition provides an elegant and simple method of covering 16 a wound site with a carbon dioxide enriched atmosphere, without requiring any specialized 17 equipment to introduce the carbon dioxide to the site of application. After applying, the 18 portion of the composition exposed to air may dry to form a thin skin that further aids in 19 sealing the carbon dioxide enriched atmosphere to the wound site. This also serves to seal the wound site from the surrounding environment, protecting it from contaminants. In the 21 case where moist wound care is desired, the composition helps to help the wound site 22 hydrated. The composition readily fills and penetrates the wound site, delivering both 23 carbon dioxide and the honey to areas of the wound that would normally be difficult to 24 access. As noted above, the honey helps to draw out wound exudate by its strong osmotic action on body tissue, and provides antimicrobial, anti-inflammatory and antioxidant effects. 26 [0049] The composition also provides a source for generating additional carbon dioxide: 27 as the composition comes in contact with moisture from body tissue, or wound exudate, the 28 excess mineral salt within the composition reacts with this moisture to form additional carbon 29 dioxide.

[0050] The composition has an adherent contact surface that allows it to bind to both 31 wet and dry wound surfaces, as well as wound dressings, which may be optionally applied 32 over the composition. As the pH of the composition can be adjusted to about 6.3 to 7.4, the 33 pH would be close to that of body tissues including epithelial and connective tissues, as well PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 as interstitial fluid. Consequently, the application of the composition to the wound site is not 2 expected to irritate or cause significant pain to the patient. The composition may be applied 3 by an unskilled practitioner, or it may be self-applied by the patient. 4 [0051] The composition may be reapplied as required during the course of wound healing. It is expected that after the initial application, the composition will eventually lose 6 activity over time as the honey will be diluted by moisture or wound exudate, the carbon 7 dioxide comprised within the composition will gradually escape to the atmosphere, and/or 8 the excess bicarbonate or carbonate salt will be eventually be consumed. 9 [0052] In an embodiment, there is provided a system comprising the above-described composition, and an isotonic diluent buffer solution. The isotonic diluent solution may be 11 used to dilute the composition to provide an irrigation solution that can be used to cleanse 12 and rinse out the wound site before fresh composition is applied. This may be done before 13 the initial application of composition is applied to the wound, or it may be done during the 14 course of subsequent applications of the composition. In an embodiment, the diluent solution comprises ascorbic acid, sodium bicarbonate, and sodium chloride. Preferably, the 16 diluent solution further comprises a surfactant and a pharmaceutically acceptable 17 preservative. The diluent solution can be used to dilute the composition by any degree, as 18 would be understood by persons skilled in the art. For example, in one embodiment, the 19 diluent solution can be used to dilute the composition to a ratio (the volume of composition to the volume of diluent solution) of about 1:2 to 1:30. Any other dilution ratios within this range 21 are also contemplated, such as 1:2, 1:5, 1:10, 1:15, 1:20, 1:25 and 1:30, inclusive of all 22 ratios between these values. Various medical devices may be employed to apply the wound 23 irrigation solution to the wound, including but not limited to: debriding sponges, pulsatile 24 wound irrigation devices, mechanical spray irrigation devices (including spray and vacuum devices), squeeze bottle dispensers and the like. 26 [0053] Besides its use in promoting the healing of chronic wounds, the composition may 27 also be used to promote the healing of other medical concerns, for example, oral ulcers, 28 surgical wounds, trauma wounds, minor cuts, abrasions, burns, puncture wounds and 29 sunburn.

[0054] In another embodiment, the composition may be used as a veterinary wound 31 care agent, to promote wound healing in wild and domesticated animals. Such wounds may 32 include lacerations, abrasions, myiasis (fly bite) ulcerations, and burns resulting from 33 chemical, thermal or electrical exposure. For such veterinary uses, the composition may PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 further comprise a suitable insect repellant such as citronella, in order to limit insect contact 2 with the wound site. 3 [0055] It has also been discovered that the above-described composition is useful for a 4 variety of cosmetic applications. For example, it may be used for superficial exfoliation of the epidermis to cosmetically improve skin appearance. 6 [0056] The composition would also be used to improve the appearance of 7 hyperpigmented areas. This can be achieved by exfoliation to remove pigmented skin cells, 8 revealing less pigmented skin below. 9 [0057] In cosmetics, sodium bicarbonate is sometimes used as a mechanical exfoliant in facial or body scrubs and it is also found in compositions that provide an antiseptic and 11 deodorant effect; in food sciences, sodium bicarbonate or sodium carbonate (both in pure, 12 solid form, or in aqueous solution) are sometimes used to break down protein, due to their 13 basic pH. However, sodium bicarbonate can be overly harsh as a mechanical exfoliant and 14 it can be irritating to the skin, and this effect would be common to any other mineral salts of carbonate or bicarbonate. As well, solutions of bicarbonate or carbonate salts are basic in 16 pH and would also be irritating or damaging to the skin. The above-described composition is 17 far superior to sodium bicarbonate alone, as it is provides a mild yet effective means of 18 superficial exfoliation of the epidermis without causing any irritation or damage to the skin. 19 As an exfoliant, the composition is more effective than either honey alone, or mineral salts of carbonate or bicarbonate. 21 [0058] The composition may also be used to enhance recovery from acne, as it provides 22 an antibacterial and anti-inflammatory effect, while at the same time, it exfoliates the 23 epidermis and helps to clear infected pores. The composition may be applied as a mask 24 that is left on the skin for a short period of time, and then rinsed off.

[0059] In comparison to other commonly used cosmetic exfoliants including mechanical 26 exfoliants (e.g. cleansers containing small particles which act as abrasives; brushes, cloths 27 and sponges), and chemical exfoliants such as alpha and beta hydroxy acids, retinoids, and 28 topical acne treatments such as benzoyl peroxide, the above-noted composition is non- 29 irritating and is well tolerated by most users, with low allergenic potential.

[0060] The composition may be also be used to manage and improve chronic skin 31 conditions including ichthyosis, psoriasis, seborrheic dermatitis, atopic dermatitis and 32 eczema. The composition forms an occlusive barrier on the skin, which provides PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 moisturizing and humectant effects. The composition aids in exfoliation of the build-up of 2 dead skin cells, a common feature in all of these skin conditions. At the same time, the 3 composition serves to calm inflammation, inhibit secondary microbial infections, and promote 4 healing of the skin.

[0061] In addition to the above, the composition may be provided in the form of a lip 6 balm or lip ointment. As a lip balm, the composition enhances exfoliation of dried skin from 7 the lips while moisturizing the lips. Both of these properties are highly useful to users in cold 8 and/or dry climates. The composition also promotes the healing of cold sores and fever 9 blisters, which are symptoms exhibited by patients suffering from other diseases or infections, such as Herpes Simplex I infection. 11 [0062] In an embodiment, the mineral salt is a pharmaceutically acceptable, water 12 soluble salt, preferably a carbonate and/or bicarbonate salt, and wherein the salt evolves 13 carbon dioxide (CO ) gas upon contact with an acid and water. In a further embodiment, the 14 one or more salts are selected from sodium bicarbonate, ammonium bicarbonate, potassium bicarbonate, calcium bicarbonate, sodium carbonate, potassium carbonate and 16 combinations thereof. In a preferred embodiment, the salt is sodium bicarbonate. 17 [0063] In another embodiment, the honey is medical grade honey. In yet another 18 embodiment, the honey is medical grade honey derived from bees that have fed primarily or 19 exclusively on plants of Leptospermum sp. In a preferred embodiment, the honey is obtained from the hives of bees that have fed exclusively on Leptospermum scoparium, 21 commonly referred to as Manuka, New Zealand tea tree, broom tea tree or simply tea tree. 22 Honey of this source is also commonly referred to as Manuka honey. 23 [0064] In an embodiment, the composition is provided in the form of a foam or mousse, 24 wherein said foam or mousse comprises bubbles or cells that further comprise carbon dioxide gas. The foam is preferably capable of generating additional carbon dioxide gas 26 upon contact with acid and moisture. In another embodiment, the composition is provided 27 as an ointment, cream, lotion, gel or balm, wherein said ointment, cream, lotion, gel, or balm 28 is capable of generating carbon dioxide gas upon contact with acid and moisture. If the 29 composition is provided in the form of an ointment, cream, lotion, gel or balm, it may further comprise bubbles or cells that further comprise carbon dioxide gas. 31 [0065] Thickening agents may be added to the composition to provide a thicker or stiffer 32 consistency, which may be desirable under various circumstances. An example would be PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 the care required for chronic wounds since, in such cases, the composition must stay in 2 place for longer periods of time. Chronic wounds include but are not limited to ulcers, 3 surgical wounds, and burns and those comprising necrotic tissue. Thickening agents may 4 also help to absorb moisture such as wound exudate, thus preventing dilution of the composition, and extending the activity of the composition. Examples of suitable thickening 6 agents include but are not limited to: polysaccharides such as starches, agar, alginic acid 7 and salts thereof (e.g. sodium alginate), carrageenan, pectin, beta glucan, and natural gums 8 such as xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum and combinations 9 thereof. In a preferred embodiment, the thickening agent is xanthan gum. Other suitable thickening or gelling agents will be known to persons skilled in the art. 11 [0066] Suitable surfactants may also be added to the composition to aid in denaturation 12 of non-viable tissue, and thus promote the debriding or desloughing effect. An example of 13 such a surfactant is sodium lauryl sulfate. In an embodiment, the composition comprises 14 about 0.0001 to 0.1% w/w sodium lauryl sulfate. Other suitable surfactants will be known to persons skilled in the art. 16 [0067] In another embodiment, the composition may further comprise one or more 17 proteolytic enzymes (proteases). The protease provides enzymatic debridement to the 18 composition and further enhances the activity of the composition for degradation and 19 removal of non-viable or necrotic body tissue. Autolytic debridement, as brought on by application of honey, is considered to be slow acting, compared to enzymatic debridement. 21 Thus, the addition of more or more proteases may extend the activity of the composition. 22 The proteolytic enzyme may be plant-derived proteases, such as papain, actinidin, 23 bromelain, ficain, or cucumisin, or bacteria-derived proteolytic enzymes. In a preferred 24 embodiment, the composition comprises actinidin. In another preferred embodiment, the composition comprises a plant extract that comprises actinidin. The plant extract is 26 preferably kiwi fruit extract, which contains actinidin. The plant proteases may be added to 27 the composition in crude or purified form. If the composition comprises one or more plant- 28 derived proteases, the composition may additionally contain antimicrobial agents such as 29 methylglyoxal or its precursor, 1,3-dihydroxyacetone.

[0068] In yet another embodiment, the one or more proteolytic enzymes, if present, are 31 preferably provided in their zymogen form. As used herein, the term "zymogen" will be 32 understood to mean an inactive form of an enzyme that is converted to its active form by PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 action of an activating agent. Such activating agent may include an acid, another enzyme or 2 a combination thereof. 3 [0069] In order to maintain desired properties and to increase storage time (shelf life) 4 over prolonged periods of time, the composition may be supplemented with one or more pharmaceutically acceptable preservatives as known in the art. Examples of preservatives 6 that may be used in the present invention include methyl paraben, propyl paraben, butyl 7 paraben, imidazolidinyl urea, tetradecyl-trimethyl ammonium bromide (Cetrimide®), sodium 8 benzoate, potassium sorbate, thymol, polyaminopropyl biguanide, and combinations thereof. 9 Various other preservatives will be known to persons skilled in the art.

[0070] As described above, in an embodiment of the present invention, the composition 11 comprises honey that is produced by bees that have fed mainly or exclusively on 12 Leptospermum plant species. Preferably, the honey is Manuka honey produced from 13 Leptospermum scoparium. In another embodiment, the Manuka honey has of a pH of about 14 3.2 to 4.5 and a minimum methylglyoxal concentration of about 150 mg/kg. In one embodiment, particularly for wound treatment, the composition of the invention comprises 16 about 80% (w/w) or more of honey. 17 [0071] In another embodiment, the composition is in the form of a foam or mousse, 18 preferably comprising cells or bubbles of carbon dioxide. The composition may also be in 19 the form of a gel, lotion, cream, ointment or balm. These other forms may further comprise cells or bubbles comprising carbon dioxide. 21 [0072] In yet another embodiment, the pharmaceutically acceptable salt is: sodium 22 carbonate (Na CO ), potassium carbonate (K CO ), ammonium carbonate ((NH ) CO ), 2 3 2 3 4 2 3 23 sodium bicarbonate (NaHCO ), potassium bicarbonate (KHCO ), ammonium bicarbonate 24 ((NH )HCO ) or any combination thereof. In one embodiment, the pharmaceutically acceptable salt is sodium bicarbonate (NaHCO ). 26 [0073] The pharmaceutically acceptable salt may be present in the composition at about 27 2.5 to 20% w/w of the composition. In one aspect of the invention, the salt is present at 28 about 5 to 15% w/w of the composition. In another aspect, the salt is present at about 7.5 to 29 12.5% w/w of the composition. In one aspect of the invention, the concentration of the salt may be varied according to the desired use of the composition. For example, if the 31 composition is for use in treating a wound with necrotic tissue and malodor, the salt (e.g. 32 sodium bicarbonate) concentration may be 12.5% -15.0%. If the composition is for use in PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 treating a wound after all necrotic tissue has been removed and is in later stages of 2 granulation, the salt concentration may be 7.5%-10%. If the composition is for use in 3 treating a wound with completed granulation and where re-epithelialization has started, the 4 salt concentration may be 2.5%-5.0%. Thus, as would be understood by persons skilled in the art, a higher salt concentration would be preferred when the composition is used in a 6 wound at an early stage of healing. However, it will also be understood that such preferred 7 embodiment of the invention is not intended to restrict the concentration of the salt 8 component based on the state of the wound. 9 [0074] In an embodiment of the present invention, the composition may further comprise one or more organic acids. For example, the organic acids may be: gluconic acid, lactic 11 acid, acetic acid, butyric acid, citric acid, malic acid, pyroglutamic acid, succinic acid, 12 ascorbic acid, glycolic acid, lactic acid, citric acid, mandelic acid or any combination thereof. 13 The one or more organic acids of the composition would be understood to be in addition to 14 those acids naturally occurring in honey. In one embodiment, the organic acid used in the composition is ascorbic acid. The organic acids added to the composition may be present at 16 a concentration of about 0.2 to 10% w/w of the composition. In one aspect, the acid is 17 present at a concentration of about 1 to 5% w/w of the composition. Generally, the 18 concentration of the organic acid would be that needed to adjust the pH of the composition 19 from about 5 to about 7.4 and, preferably, from about pH 5.5 to about 6.5. More preferably, the pH of the composition is adjusted to be about 7 to 7.4. As would be understood, where 21 the concentration of the carbonate and/or bicarbonate salt is high, resulting in a greater 22 amount of CO release, a higher concentration of the organic acid would be needed to adjust 23 the pH of the composition to the aforementioned desired levels. 24 [0075] In another embodiment of the present invention, the composition may further comprise one or more thickening agents. For example, the thickening agents may be: 26 polysaccharides, starches, agar, alginic acid and salts thereof, carrageenan, pectin, beta 27 glucan, xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, or any 28 combination thereof. In one embodiment, the composition comprises xanthan gum. The 29 thickening agents may be present at a concentration of about 0.2 to 6% w/w, with the concentration being dependent upon the desired viscosity of the composition. For example, 31 a sufficient amount of thickening agent may be incorporated to provide a viscosity, at skin 32 temperature, that is sufficient to cover and seal the site of application (e.g. the wound area) 33 so as to provide an environment enriched in CO and reduced in O . In one aspect, the PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 thickening agents may be present at a concentration of about 1 to 5% w/w. In another 2 aspect, the thickening agents may be present at a concentration of about 2 to 5% w/w. 3 [0076] In yet another embodiment, the composition may further comprise sodium lauryl 4 sulfate. Sodium lauryl sulfate may present in the composition at a concentration of about 0.0001 to 0.1% w/w. 6 [0077] In another embodiment, the composition further comprises one or more 7 proteolytic enzymes. For example, the proteolytic enzymes can be: papain, actinidin, 8 bromelain, ficain, cucumisin, bacteria-derived proteolytic enzymes, or any combination 9 thereof. The enzyme may be provided as an extract of the plant or bacteria that comprises one or more of the above-noted enzymes. In a further embodiment, the enzyme is actinidin. 11 Actinidin may be provided in kiwi fruit or an extract thereof. Such an extract includes juice of 12 the kiwi fruit, or a concentrate thereof. 13 [0078] In yet another embodiment, the composition further comprises one or more 14 preservatives. For example the preservatives may be: methyl paraben, propyl paraben, butyl paraben, imidazolidinyl urea, tetradecyl-trimethyl ammonium bromide (Cetrimide®), 16 sodium benzoate, potassium sorbate, thymol, polyaminopropyl biguanide, or any 17 combination thereof. 18 [0079] In an embodiment of the present invention, the composition has an osmotic 19 concentration at least 14.5 times the osmotic concentration of human blood.

[0080] In another aspect of the present invention, there is provided a use of the 21 composition as described herein, as a desloughing agent or a debriding agent, a wound 22 treatment agent, a veterinary wound care agent, and for the treatment of chronic skin 23 disorders. Such skin disorders may comprise, for example: ichthyosis, psoriasis, seborrheic 24 dermatitis, atopic dermatitis, and eczema. There is also provided a use of the composition as a cosmetic skin exfoliant. 26 [0081] In yet another aspect of the present invention, there is provided a system of 27 wound treatment comprising the composition according to any of the above-described 28 embodiments and an isotonic diluent solution, preferably of about pH 7, comprising: about 29 0.015 M ascorbic acid; about 0.015 M sodium bicarbonate; and, about 0.12 sodium chloride;

[0082] - wherein the isotonic diluent solution is used to dilute said composition at a 31 minimum ratio of 1 part said composition to 2 parts said diluent solution (1:2), thereby PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 forming a wound irrigation solution; and wherein the wound irrigation solution is used to 2 irrigate a site of treatment on a patient before application of said composition. 3 [0083] An advantage of the invention is that is provides an unexpected, and significant 4 enhancement of the removal of non-viable tissue, that is far greater than observed for each -2 -1 of honey, carbon dioxide, and mineral salt of carbonate (CO ) or bicarbonate (HCO ). The 6 enhancement of wound healing and therapeutic effect is also far greater than would be 7 expected from a simple additive effect of honey, carbon dioxide, and mineral salt of 8 carbonate or bicarbonate, used separately to treat a wound site. 9 [0084] Another advantage of the invention is that it may be used as a desloughing or debriding agent, for the purposes of promotion of wound healing, or it may be for superficial 11 exfoliation of the epidermis to cosmetically enhance skin appearance. 12 [0085] Yet another advantage of the invention is that it provides an elegant and simple 13 method of covering a wound site with a carbon dioxide enriched atmosphere, and honey, 14 both of which provide numerous beneficial effects in the promotion of wound healing, including but not limited to: 16 [0086] - providing a hyperosmotic environment (also referred to as an environment of 17 negative osmotic pressure) that draws out and absorbs wound exudate; 18 [0087] - providing a means of stimulating oxygen and microcirculation to the wound site; 19 and,

[0088] - providing a carbon dioxide-enriched micro-climate over the wound that 21 discourages growth of aerobic bacteria. 22 [0089] Another advantage of the invention is that it is mild, with a pH that can be 23 adjusted close to the pH of interstitial body fluids and body tissues including epithelial and 24 connective tissues; consequently, the application of the composition to the wound site is not expected to irritate or cause significant pain to the patient. 26 [0090] Yet another advantage of the invention is that it is simple and does not require 27 specialized equipment to use or apply: the composition may be applied by an unskilled 28 practitioner, or it may be self-applied by the patient. 29 [0091] Another advantage of the invention is that it may be used on human patients or it may also be used to promote wound healing in wild or domesticated animals.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [0092] Another advantage of the invention is that it may be used for superficial 2 exfoliation of the epidermis to cosmetically improve skin appearance. It may also be used to 3 enhance recovery from acne, as it provides an antibacterial and anti-inflammatory effect, 4 while at the same time, it exfoliates the epidermis and helps to clear infected pores. In addition, the invention may be also be used to manage and improve chronic skin conditions 6 including ichthyosis, psoriasis, seborrheic dermatitis and eczema. 7 [0093] In all cases, an advantage of the invention is that it is much milder than known 8 treatment methods, with low irritation and allergenic potential, while still providing effective 9 treatment and management of the above-noted medical concerns.

[0094] Other and further advantages and features of the invention will be apparent to 11 those skilled in the art from the following detailed description of an embodiment thereof, 12 taken in conjunction with the accompanying drawings. 13 [0095] Kits 14 [0096] In one aspect, there is provided a treatment kit comprising the honey component and the carbonate and/or bicarbonate salt component of the composition, as discussed 16 above. Optionally, the kit may include any necessary instructions for forming and using the 17 composition and/or any variety of containers or vials etc. for mixing the components to form 18 the composition. 19 [0097] In one aspect, the honey component of the composition is provided in dry or powder form. Similarly, the carbonate and/or bicarbonate salt component may also be 21 provided in dry or powder form. As would be understood, providing the components of the 22 invention in dry form allows for extended shelf life. In the case of dried or powdered honey 23 and dried or powdered carbonate/bicarbonate salt(s), the two components can be provided 24 in a kit comprising one or two containers or packages for the honey, carbonate and/or bicarbonate salt, or a mixture thereof. In such case, the kit may further comprise a container 26 or package containing a volume of water, which can be used to form the composition 27 described above. As mentioned above, the kit may include a container into which the 28 dry/powder components are combined with the water. In one example, the container may 29 contain the necessary volume of water and the dry honey and salt components may be provided in a package or sachet. Alternatively, the container may be provided in an empty 31 state, prepared to receive the dry/powder components and the required volume of water 32 from an acceptable source.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [0098] In another aspect, the dried honey and carbonate/bicarbonate salt components 2 can be combined into a dressing or other such application medium. Once wetted, the 3 components of the composition would become wetted and thereby activated. In one aspect, 4 the dressing would be adapted to form a foam or mousse as described above. The wetting of the dressing can be achieved by adding water or by using the natural exudate originating 6 from a wound. 7 [0099] In one aspect, the above mentioned dressing may incorporate coated honey 8 crystals, which are activated by the high pH of the bicarbonate once hydrated by added 9 water or would excretions. Such coating may be provided by microencapsulation of the dried honey. 11 [00100] In another aspect, the dried honey, the carbonate/bicarbonate salt and the acid 12 components can be combined together in dry form in a single airtight pouch. In one aspect, 13 the honey may be coated (or microencapsulated) to allow for a release under certain 14 conditions. In another aspect, neither of the ingredients would need to be coated as they would be inert in their dry state. The powder composition, such as the non-coated particles, 16 can then be applied to a region to be treated, such as a skin ulcer, and then covered to 17 allow the treatment phase to take place. 18 [00101] In the above description, the use of natural honey, in either liquid or dry (and/or 19 encapsulated) form, has been discussed. However, in further aspects, such natural honey can be replaced with a synthetic honey or a honey substitute. For example, corn syrup of a 21 suitable viscosity can be used instead of honey to entrap carbon dioxide released at the 22 point of use due to the reaction of organic acid(s) and sodium bicarbonate. Similarly, 23 suitable compositions of invert sugars such as corn syrup of such viscosity and low water 24 content as to approximate honey (86% sugars) in a ratio of 55% fructose and 45% glucose may be used as alternatives to honey in necrotic wound and healing formulations as 26 described above. 27 [00102] Examples 28 [00103] Aspects of the invention will now be described with reference to various 29 examples. It will be understood that the following examples are provided to illustrate the present invention and are not intended to limit the scope of the invention in any way.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00104] Example 1: Preparation and Testing of Treatment Compositions 2 [00105] (i) Preparation of Treatment Compositions 3 [00106] The following compositions were prepared. Medical grade Manuka honey 4 derived from Leptospermum scoparium (Links Medical LLC) was used to prepare the compositions, wherein the medical grade honey had a pH 3.63, and was defined as having 6 an activity equivalent to 16% phenol, also referred to as level 16 on the 20 point Unique 7 Manuka Factor (UMF) scale. Either sodium bicarbonate or potassium bicarbonate could be 8 used in the formulations noted below. The results shown below are for formulations 9 prepared with sodium bicarbonate.

[00107] Table 1: Compositions of honey and sodium bicarbonate (% w/w of total) Formulation Honey Sodium bicarbonate pH As noted in Figure 1 (% w/w) (NaHCO ) (% w/w) A 97.5 2.5 6.35 HoneyMousse 2.5% B 95.0 5.0 6.95 HoneyMousse 5% C 92.5 7.5 7.0-7.2 HoneyMousse 7.5% D 90.0 10.0 7.0-7.2 HoneyMousse 10% E 87.5 12.5 7.2 HoneyMousse 12.5% Control 100.0 0 3.63 Honey Control 12 [00108] Upon addition of sodium bicarbonate to honey, a composition in the form of a 13 stable foam was formed, comprising minute cells, each cell enclosing a carbon-dioxide 14 enriched atmosphere.

[00109] (ii) In Vitro Testing of Debriding/Desloughing Efficiency 16 [00110] Figure 1 shows the debriding or desloughing efficiency of each of the above 17 compositions, when applied to a sample of pig skin in vitro, at room temperature. Since the 18 melanin pigment in pig skin is located primarily in the basal layer of the epidermis, 19 detachment of the dark-coloured epidermis reveals the amber-coloured dermis beneath.

Therefore, detachment of the epidermis may be easily visualized and was taken as a marker 21 of successful debriding or desloughing of the epidermis from the dermis. Squares of pig 22 skin, 2.5 cm2 in area, were treated with each of formulations (A) to (E) of Table 1, according 23 to the following protocol, repeated every 12 hours: (1) wash with water, (2) photograph and PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 quantify debrided area and (3) reapply 1.5 g of formulation to each square of pig skin (i.e. 2 approximately 0.24 g per 1 cm2). The area of the revealed dermis was quantified on a grid 3 and calculated as a percentage of the total area of the sample, at each 12 hour application 4 point.

[00111] As shown in Figure 1, there was an incremental increase in debriding or 6 desloughing efficiency, dependent on sodium bicarbonate concentration, which reached a 7 plateau at around 12.5% w/w sodium bicarbonate, in Formulation (E). 8 [00112] Formulation (E) was then compared with the debriding efficiency of commercially 9 available debriding compositions, Santyl collagenase ointment (Smith & Nephew, Inc.) and Kovia papain-urea ointment (521,000 units papain activity per gram ointment; Stratus 11 Pharmaceuticals Inc.). Debriding/desloughing efficiency was measured in vitro on pig skin, 12 according to the same experimental protocol as described above. Figure 2 shows the 13 results of the comparison of in vitro debriding efficiency of Formulation (E), containing 12.5 14 % w/w sodium bicarbonate, compared with commercially available debriding compositions, Santyl collagenase ointment (Smith & Nephew, Inc.) and Kovia papain-urea ointment (as 16 above; Stratus Pharmaceuticals Inc.). 17 [00113] As shown in Figure 2, Formulation (E) comprising 12.5% w/w sodium bicarbonate 18 provided faster and more effective debridement of the pig skin, compared to either Kovia 19 papain-urea ointment or Santyl collagenase ointment.

[00114] (iii) In Vivo Tests of Debriding/Desloughing Efficiency 21 [00115] (a) Treatment of equine myiasis ulcer on donkey 22 [00116] A donkey with a large myiasis ulceration on lateral aspect of right front knee was 23 treated with Formulation (E). 24 [00117] The ulceration wound was 8.6 cm wide by 9.4 cm high and presented with heavy encrustation of blood. The wound was cleansed with 3% hydrogen peroxide and irrigated 26 with sterile saline on Day 0 and was treated to debride and heal by secondary intention. The 27 study consisted of daily removal of the dressing and saline irrigation followed by application 28 of the following at the given time periods. 29 [00118] Day 0: Pre-Debriding Ointment (as provided in U.S. Patent Application Publication No. 20130085461) was applied for 24 hours. Ambient temperature was 32- PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 35 C. Dressed with non-stick dressing and wrapped with leg pad and held with cloth strap 2 and Velcro® closure. 3 [00119] Days 1-7: Kiwi fruit juice concentrate (containing naturally-occurring actinidin) 4 plus formulation A (Leptospermum scoparium (Manuka) honey and 2.5% sodium bicarbonate, pH 6.35); 6 [00120] Days 8-29: Medical grade Manuka Leptospermum scoparium honey only, pH 7 3.63 (Links Medical LLC); 8 [00121] Days 30-68: Formulation E (Leptospermum scoparium honey and 12.5% sodium 9 bicarbonate, pH 7.2)

[00122] The composition of the Pre-Debriding Ointment was as follows (also as provided 11 in U.S. Patent Application Publication No. 20130085461): Ingredient Concentration (% W/W) Urea 25.00 Waxes and mineral oils 10.00 6.00 PEG-100 (Carbowax ) lidocaine hydrochloride 4.00 n-propanol 4.00 lanolin 4.00 cetyl alcohol 3.00 glucose (USP grade) 2.00 mandelic acid 2.00 13 [00123] During debriding (days 1-7), treatment was daily. After debriding during 14 granulation and remodeling of granulation tissue (days 8-29), treatment was every 3 to 4 days. During dermal and epidermal tissue construction and migration, to wound closure, the 16 treatment composition was applied every 4 to 7 days. Photographs were taken during 17 treatment; Figure 3 shows photographs taken on days 0, 7, 30 and 68 of the treatment. 18 Observations at the end of each stage of treatment are as provided below. 19 [00124] Following the pre-debriding ointment treatment for 24 hours at 32-35 C on heavy eschar and necrotic tissue, the subsequent use of kiwi fruit juice enzyme (actinidin) in honey PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 mousse was able to debride the wound in 4 days and promote rapid granulation. Debriding 2 was continued with the treatment composition to day 7 to ensure complete cleansing. At day 3 7, the wound appeared to be vigorously granulating with good vascularity and has filled the 4 wound deficit. Medical grade Manuka (Leptospermum scoparium) honey alone was used on days 8 to 28, and produced good granulation development and remodeling up to Day 30. At 6 day 30, epithelial migration continued to close the wound. Granulation tissue is also 7 continuing to remodel and vascularize and innervate. 8 [00125] Formulation (E) from Table 1 continued the healing with epidermal migration 9 starting around day 30. Formation of new epithelium was about 75% complete at 49 days, about 90% complete at 63 days, and about 93% complete at 68 days. This rate of 11 epithelialization appeared to be slightly delayed and indicates that the honey mousse (12.5% 12 sodium bicarbonate) may be best suited for the debriding and granulation phase of wound 13 healing. 14 [00126] (b) Treatment of equine myiasis ulcer on donkey

[00127] A donkey with ankle wounds was presented for treatment. Myiasis ulcerations 16 2.5 cm x 1.5 cm were located on the anterior aspect of the front right ankle. The donkey had 17 been plagued by fly bite irritated skin resulting in ulceration and erosion of epidermis and 18 dermis. Left uncovered and untreated, this ulceration would enlarge rapidly as did the 19 ulceration on his knee above.

[00128] The affected area was treated with Formulation (A) and then Formulation (E), 21 according to the following protocol. Photographs were taken during treatment; Figure 4 22 shows photographs taken on days 0, 3 and 18 of the treatment. Observations at the end of 23 each stage of treatment are as provided below. 24 [00129] Day 0: Pre-Debriding Ointment (as noted above in Example 1(iii)(a); as provided in U.S. Patent Application Publication No. 2013/0085461) was applied and left in place for 24 26 hours. Ambient temperature was 35 C. 27 [00130] Day 1-3: Kiwi fruit juice with actinidin enzyme, 40 units/g, was added to 28 formulation (A) (Manuka Leptospermum scoparium honey and 2.5% w/w sodium 29 bicarbonate), and an amount sufficient to cover wound was applied to the wound site. For 3 days, the wound was washed with saline and the non-stick dressing was changed every 24 31 hours. The primary dressing was with cotton wrap with Velcro closure. Debriding was PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 considered complete on day 3. The development of new granulation tissue was observed 2 on day 3. 3 [00131] Day 4-18: The wound site was treated with Formulation (E) (Manuka 4 Leptospermum scoparium honey and 12.5% sodium bicarbonate). Honey mousse left in place for 3-4 days at a time. On day 18, the wound was observed as being fully closed. 6 [00132] (c) Treatment of equine myiasis ulcer on horse 7 [00133] A horse with a large girth (chest) wound from myiasis (fly bite) ulcerations 8 measuring 10 cm x 9.5 cm x 1.2 cm deep, was presented for treatment. The wound site was 9 surgically debrided and treated with Formulation (E) (Manuka honey and 12.5% w/w sodium bicarbonate) until closure according to the following protocol. 11 [00134] The horse was given appropriate sedatives and was then positioned on its back. 12 The chest wound, found just distal to the front legs, was cleansed with betadine, and the 13 area surrounding the wound site was shaved. The necrotic tissue mass was surgically 14 excised down into the grey fibrous connective tissue over the sternum, approximately 1.5cm deep. The wound site was irrigated with sterile saline and Formulation (E) was applied to 16 non-stick pads and placed on the wound. A secondary dressing of a disposable diaper was 17 used to further secure Formulation (E) in place. On subsequent dressing changes, the 18 wound was irrigated, photographed and Formulation (E) reapplied on non-stick dressings 19 held in place with a custom-made sling created by the owner which kept the dressing secure for 3-7 days at a time. Photographs were taken during treatment; Figure 5 shows 21 photographs taken on days 0, 16, 21 and 32 of the treatment. Observations at the end of 22 each stage of treatment are as provided below. 23 [00135] Formulation (E) was effective for debriding the wound margin tissue that became 24 necrotic after the surgical excision, and was a strong stimulator of granulation and remodeling. By day 16, the wound had finished granulation and was remodeling well out to 26 the edges. By day 21, epithelial migration had started and it was decided to continue 27 application of Formulation (E) to wound closure, in order to get a second assessment of its 28 effect during this phase. The wound closed on day 32, well within normal time frame. The 29 new epithelium appeared soft and loosely organized, as observed during the treatment described in Example 1(iii)(a).

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00136] (d) Treatment of surgical excision wound on human male patient 2 [00137] A human patient, male, age 69 years, in good general health, was presented with 3 a skin pathology appearing to be a classic keratoacanthoma on his right dorsal forearm. The 4 keratoacanthoma lesion was approximately 1 cm in diameter with a central crater, raised, with no erythema. A shave biopsy was taken of the raised portion, approximately 1.4 x 1.3 x 6 0.2 cm in size, with an eccentric 0.9 x 0.7 cm white and brown lesion. At excision, the 7 capillary bleeding was managed with electrocautery. The shave biopsy specimen was sent 8 out for pathology analysis. 9 [00138] The surgical wound was closed by secondary intention and treated daily with Formulation (E) starting the day after the excision. The wound cavity was filled with 11 Formulation (E) and covered with an island type secondary bandage. At each daily dressing 12 change, the wound was irrigated and re-dressed with Formulation (E). The treatment was 13 halted on day 7, when the pathology analysis found that the biopsy as comprising squamous 14 cell carcinoma, invasive and well differentiated, present at the deep margin. Photographs were taken during treatment; Figure 6 shows the photographs taken on days 0, 5 and 7 of 16 the treatment. Observations at the end of each stage of treatment are as provided below. 17 [00139] On day 0, shave biopsy and electrocautery produced a surgical excision wound 18 site. The wound site was dressed with triple antibiotic ointment by the attending physician 19 and covered with non-stick pad with paper tape. On day 4, formation of granulation was proceeding and the wound site appeared to be approximately half filled. By day 6, the 21 granulation phase had completely filled the wound site, and the inflammatory response has 22 subsided. By day 7, it was observed that rapid debriding of cautery point necrotic tissue had 23 occurred, and granulation tissue had filled the wound site, with indications of angiogenesis. 24 [00140] The patient stated that there was no discomfort with the use of Formulation (E) from a very slight tingling sensation starting about 10 minutes after application and lasting 26 about 10 minutes during the first two applications. This minimal sensation compared 27 favorably against the stinging and burning sensations experienced by users of commercially- 28 available enzymatic debriding compositions such as papain-based debriding ointments. 29 [00141] Example 2: System for Wound Treatment

[00142] As an example, Formulation (E) of Example 1 was diluted with an isotonic diluent 31 buffer solution as follows: PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00143] Table 2 (a): Composition of Isotonic Diluent Buffer Solution Buffer Solution Ingredient Concentration Ascorbic acid 0.015 M Sodium bicarbonate (NaHCO ) 0.015 M Sodium chloride (NaCl) 0.12 M 3 [00144] Undiluted Formulation (E) has an approximate osmolarity (also referred to as 4 osmotic concentration) of 14.5 times greater than human blood. Formulation (E) of Example 1 was diluted with the isotonic diluent solution of Table 2(a), at the following dilution factors 6 provided in Table 2(b) below, with the corresponding resultant osmolarity compared to 7 human blood. 8 [00145] Table 2 (b): Dilution Factors Dilution factor Resultant hyperosmolarity of diluted (Formulation (E) : Diluent solution) formulation compared to human blood 1 : 2 7.2 x 1 : 5 3.8 x 1 : 10 2.3 x 1 : 20 1.46 x 1 : 30 1.15 x

[00146] As noted above, the hyperosmolarity of the resultant solution was the osmotic 11 concentration of the resultant solution when compared to human blood. For example, a 12 solution diluted 1:2 resulted in a solution that had an osmotic concentration 7.2 x the osmotic 13 concentration of human blood. 14 [00147] To each of these diluted solutions of Table 2(b), the following compounds were added to provide the final finished wound irrigation solutions. 16 [00148] Table 2(c): Additives to diluted solution Additive Final concentration in solution Sodium lauryl sulfate 10 mM polyaminopropyl biguanide 0.00003% w/w PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 Disodium EDTA 0.025% w/w 2 [00149] The resultant diluted compositions were used as wound irrigation solutions, i.e. to 3 irrigate the wound before applying a formulation of Example 1 (undiluted) for the first time to 4 a wound site), or while replacing applications of the formulations of Example 1 to a wound site with an application of fresh formulation. 6 [00150] Various medical devices may be employed to apply the wound irrigation solution 7 to the wound, including but not limited to, monofilament debriding sponges, pulsatile wound 8 irrigation devices, mechanical spray irrigation devices (including spray and vacuum devices), 9 and squeeze bottle dispensers.

[00151] Example 3: Cosmetic Formulations 11 [00152] (i) Sample Preparation 12 [00153] The following compositions, (a) and (b), for use as cosmetic formulations, were 13 prepared. 14 [00154] (a) Moisturizing lotion with 12% w/w sodium bicarbonate and 5% w/w honey

[00155] Each of phases A, B, C and D were blended to homogeneity and heated to the 16 specified temperature range before final blending of all phases.

Phase Ingredient %, W/W Weight (g) Phase A Purified water 48.0 480 (70-75 C ) Carbopol 940 1.5 15 Mandelic acid 1.5 15 Potassium sorbate 0.2 2 Sodium benzoate 0.2 2 Disodium EDTA 0.1 1 Glycerin 5.0 50 Urea 10.0 100 Sodium bicarbonate 12.0 120 Phase B Isopropyl myristate 5.0 50 PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 (75-80 C) Cetyl alcohol 3.0 30 PEG-100 stearate 2.0 20 Phase C Purified water 5.0 50 ( 45-50 C ) Honey (Manuka, medical grade or regular) 5.0 50 Phase D Benzyl alcohol 0.5 5 ( 35-40 C ) Silk amino acids 1.0 10 TOTAL : 100.0 1000.000 Standard pH : 8.50 2 [00156] (b) Moisturizing lotion with 5% w/w sodium bicarbonate and 5% w/w honey 3 [00157] Each of phases A, B, C and D were blended to homogeneity and heated to the 4 specified temperature range before final blending of all phases.

Phase Ingredient %, W/W Weight (g) Phase A Purified water 54.5 545 (70-75 C) Carbopol 940 1.5 15 Mandelic acid 2.0 20 Potassium sorbate 0.2 2 Sodium benzoate 0.2 2 Disodium EDTA 0.1 1 Glycerin 5.0 50 Urea 10.0 100 Sodium bicarbonate 5.0 50 Phase B Isopropyl Myristate 5.0 50 (75-80 C) Cetyl alcohol 3.0 30 PEG-100 Stearate 2.0 20 Phase C Purified water 5.0 50 (45-50 C) Honey (medical grade, Manuka or regular) 5.0 50 PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 Phase D Benzyl alcohol 0.5 5 (35-40 C) Silk amino acids 1.0 10 TOTAL : 100.0 1000 Standard pH : 7.57 2 [00158] (ii) In Vivo Test 3 [00159] A female patient suffering from severe ichthyosis was a test subject. Previously, 4 she had been using a urea and alpha hydroxy acid skin lotion (Extra Strength Body Lotion, Dermal Therapy Inc.) for several years, in order to manage the symptoms of ichthoysis. The 6 patient applied approximately 2 g of the moisturizing lotion as provided in Example 2(i)(a) 7 above, comprising 12% sodium bicarbonate and 5% honey, to an affected area on one arm, 8 twice daily, for a period of two weeks. She observed a comparable improvement to her 9 symptoms as with the urea-lactic acid cream.

[00160] Although the invention has been described with reference to certain specific 11 embodiments, various modifications thereof will be apparent to those skilled in the art 12 without departing from the purpose and scope of the invention as outlined in the claims 13 appended hereto. Any examples provided herein are included solely for the purpose of 14 illustrating the invention and are not intended to limit the invention in any way. Any drawings provided herein are solely for the purpose of illustrating various aspects of the invention and 16 are not intended to be to limit the invention in any way. The disclosures of all prior art 17 recited herein are incorporated herein by reference in their entirety. 18 [00161] Example 4: Wound Dressing Incorporating Dry Components 19 [00162] This example serves to illustrate a wound dressing incorporating Manuka honey in the form of freeze dried granules, sodium bicarbonate powder and ascorbic acid crystals. 21 [00163] Method 22 [00164] 1) Saturate sodium bicarbonate in high fructose corn syrup at 40 °C and saturate 23 the dressing. Dry with 8-ply rayon/polyester layer of dressing on non-stick conveyor belt with 24 cellulose coated layer on top, using warm (40 °C) air until completely dry and binding the two layers together.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00165] 2) Blend ascorbic acid crystals and Manuka honey freeze dried granules (pH 2 3.6-4.5) in either a debriding or healing formulation proportion and coat with Eastman™ 3 Cellulose Acetate Phthalate (CAP) using acetone:water (97%:3%) as the CAP solvent. 4 Spread the wet slurry of honey granules and ascorbic acid crystals onto “cellulose tissue” dressing surface, and allow to evaporate to dryness under negative air flow. Sodium 6 bicarbonate is less than 0.2% soluble in acetone, so almost none will be dissolved. 7 [00166] 3) When dried, seal the dressing in a typical dressing sleeve and irradiate to 8 sterilize. 9 [00167] More specifically, the method of this example involves the following. Micro- encapsulate freeze-dried Manuka honey granules blended with ascorbic acid crystals 11 (pH3.6-4.5) in Cellulose Acetate Phthalate (CAP) and adhere the encapsulated mixture to a 12 cellulose tissue layer of a gauze dressing. CAP is typically used for pharmaceutical 13 encapsulation to produce an enteric coating that resists dissolution when in an acid 14 environment, but dissolves when exposed to neutral to alkaline solution. Adhere sodium bicarbonate to the outer rayon/polyester layer of the gauze dressing. Since the wound fluid 16 is absorbed by the dressing, the dissolving sodium bicarbonate solution (pH 7-8) will cause 17 dissolution of the CAP coating and release the honey and ascorbic acid. The honey is 18 extremely hygroscopic and will immediately absorb and rehydrate with wound fluid water to a 19 honey consistency (about 14% water). The organic acids in the honey and ascorbic acid will react in the wound fluid water with dissolved sodium bicarbonate to release carbon dioxide 21 which infuses the now fluid honey to create a foam. 22 [00168] Excess carbon dioxide is released from the non-occlusive gauze dressing. 23 Occlusive adhesive border dressings should be avoided as they could inflate or even 24 dislodge with carbon dioxide pressure.

[00169] The advantage of the sodium bicarbonate-fructose:glucose impregnation of the 26 dressing is that it binds the sodium bicarbonate to the dressing (thereby preventing the 27 powder from sifting through the dressing fabric) and it presents a saturated sodium 28 bicarbonate solution in wound fluid to the necrotic tissue in the wound for maximum 29 loosening, softening and detachment activity. The undissolved sodium bicarbonate in the dressing acts as a reserve to continue to dissolve over time just as it does in fluid honey 31 model. The CAP coating of the hygroscopic honey granules and ascorbic acid causes 32 same to be adhered to the dressing and prevents them from pre-maturely absorbing water 33 and ensuring a longer shelf-life even with normal dressing enclosures. The CAP coating PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 withstands prolonged contact with acidic environments (the acidic honey and ascorbic acid), 2 then dissolves on contact with slightly acidic (pH 6.2) to neutral (pH 7-8) solutions. 3 [00170] Initially, the high fructose corn syrup/sodium bicarbonate gel produces negative 4 osmotic pressure as it hydrates, drawing in even more wound fluid and atmospheric moisture. Then the sodium bicarbonate dissolved in the wound fluid (pH 7-8) dissolves the 6 CAP coating and releases honey and ascorbic acid. The freeze dried honey granules then 7 hydrate and increase the negative osmotic pressure incrementally. Because the sodium 8 bicarbonate reaction with acid is driven by the evolving carbon dioxide gas, the equilibrium is 9 forced strongly in the direction of continued reaction until all the available sodium bicarbonate in the expanding “wet” area of the dressing is used up. The mechanism is self- 11 regulating based on available wound fluid. This mechanism will work for both the “Debriding 12 and the Healing” formulations. 13 [00171] Example 5: Negative Osmotic Pressure Dressing for Treating Necrotic 14 Wounds

[00172] Description: This dressing, or medical device, referred to as MANUKA–NW ,is 16 used to soften and facilitate removal of necrotic wound tissue using MANUKA honey (100% 17 Leptospermum scoparium honey from New Zealand). MANUKA-NW is super-saturated with 18 sodium bicarbonate for necrotic tissue detachment, and pH adjusted to neutral with Vitamin 19 C to optimize autolytic debriding enzyme activity. MANUKA-NW Activator (sodium bicarbonate and Vitamin C ) is mixed with MANUKA honey at Point-Of-Use to generate 21 carbon dioxide which foams the viscous honey. The high sugar level of MANUKA-NW 22 provides Negative Osmotic Pressure to draw wound fluid and autolytic cleansing to the 23 wound. 24 [00173] The following are some benefits of the activated MANUKA-NW :

[00174] 1) Improves removal and application at dressing changes, since it is less sticky 26 than honey. 27 [00175] 2) Absorbs and/or neutralizes wound malodors with sodium bicarbonate., 28 [00176] 3) Expands into difficult-to-reach undermined or tunneled necrotic areas of the 29 wound to provide moist wound environment.

[00177] 4) Provides neutral pH needed for optimal autolytic debridement.

PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00178] 5) Provides a temporary CO rich atmosphere over the wound to discourage 2 growth of aerobic bacteria. 3 [00179] 6) Manuka honey supersaturated with sodium bicarbonate rapidly loosens, 4 softens and detaches necrotic epidermis and dermis tissue.

[00180] The Activated MANUKA NW is applied to the wound containing necrotic tissue 6 and wound debris, covered with gauze (which may be supplied as part of the kit), then 7 secured in place with a suitable secondary dressing or wrap. 8 [00181] The contents of the kit could preferably comprise: 9 [00182] 1) 0.5oz. (14g) sterile 100% Manuka (Leptospermum scoparium) Honey in a flexible tube with foil seal under threaded cap. 11 [00183] 2) Activator: 0.55g ascorbic acid and 2.35g sodium bicarbonate powder in sealed 12 container. 13 [00184] Example 6: Negative Osmotic Pressure Dressing for Healing Wounds 14 [00185] Description: This dressing, or medical device, referred to as MANUKA N.O.P.D. is primarily designed for healing wounds and ulcers using Manuka Honey (100% 16 Leptospermum scoparium Honey from New Zealand) in a sterile container, activated at 17 point-of-use with ascorbic acid (Vitamin C) and sodium bicarbonate powder to create an 18 expanding, pH adjusted (6.2), carbon dioxide (CO2) infused, honey with Negative Osmotic 19 Pressure. The activated MANUKA N.O.P.D. is applied to a 4 in. X 4 in. foam pad dressing (included), then secured in place with a suitable secondary dressing or wrap. 21 [00186] Activated Manuka honey’s high sugar level provides Negative Osmotic Pressure 22 that promotes continuous autolytic debridement and helps maintain a moist environment 23 conducive to wound healing. Sodium bicarbonate rapidly absorbs and neutralizes wound 24 malodors. The CO infused consistency of the activated honey provides:

[00187] The benefits of this device include: 26 [00188] 1) Improved application and removal; 27 [00189] 2) Expansion into undermined or tunneled areas of the wound without trauma or 28 discomfort to the patient; and PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 [00190] 3) Temporary CO rich atmosphere over the wound to discourages growth of 2 aerobic bacteria. 3 [00191] This medical device would contain: 4 [00192] 1) 0.5oz. (14g) sterile 100% Manuka (Leptospermum scoparium) Honey in a flexible tube with foil seal under threaded cap; 6 [00193] 2) Activator: 1.8g ascorbic acid and 0.6g sodium bicarbonate powder in sealed 7 container. 8 [00194] Example 7: Use of Manuka Honey Freeze Dried Granules Instead of Fluid 9 Honey in Wound Dressings

[00195] Freeze-dried Manuka Honey granules were microencapsulated and blended with 11 ascorbic acid crystals (pH 3.6 - 4.5) in Cellulose Acetate Phthalate (CAP) and the 12 encapsulated mixture was adhered to the cellulose tissue layer of a gauze dressing. CAP is 13 typically used for pharmaceutical encapsulation to produce an enteric coating that resists 14 dissolution when in an acid environment, but dissolves when exposed to neutral to alkaline solution. 16 [00196] Sodium bicarbonate was adhered to the outer rayon/polyester layer of the gauze 17 dressing. As wound fluid is absorbed by the dressing, the dissolving sodium bicarbonate 18 solution (pH 7-8) will cause dissolution of the CAP coating and reveal the honey and 19 ascorbic acid. The honey is extremely hygroscopic and will absorb and rehydrate with wound fluid water to a honey consistency (about 14% water). The honey organic acids and 21 ascorbic acid will react in the wound fluid water with dissolved sodium bicarbonate to release 22 carbon dioxide which infuses the now fluid honey to create the foam of this present patent 23 application. 24 [00197] Excess carbon dioxide is released from the non-occlusive gauze dressing.

Occlusive adhesive border dressings should be avoided as they could inflate or even 26 dislodge with carbon dioxide pressure. 28 [00198] Although the above mentioned proportions of honey granules to ascorbic acid are 29 preferably suited for use in a wound healing or treating formulation, a similar formulation can be used for the other uses mentioned in the present description. For example, in the case of PCT Application Blakes Ref: 50586/00060 Doc No: 22793025.2 1 a debriding formulation, the amount of the sodium bicarbonate component would be 2 preferably higher while the proportion of the ascorbic acid would be less. 3 [00199] References 4 [00200] 1. Irish, J., Blair, S., Carter D.A. “The Antibacterial Activity of Honey Derived from Australian Flora.” PLoS ONE. 2011, 6(3): e18229. 6 [00201] 2. Kwakman, P. H. S. et al. “Two Major Medicinal Honeys Have Different 7 Mechanisms of Bactericidal Activity.” PLoS ONE. 2011, 6(3): e17709. 8 [00202] 3. Atrott, J., Henle, T. “Methylglyoxal in Manuka honey – correlation with 9 antibacterial properties.” Czech Journal of Food Sciences. 2009, 27: S163-S165.

[00203] 4. Smith, A. M., Hunt, N.C., Shelton, R. M., Birdi, G., and Grover, L.M. “Alginate 11 Hydrogel Has a Negative Impact on In Vitro Collagen Deposition by Fibroblasts.” 12 Biomacromolecules. 2012, 13(12): 4032-4038. 13 [00204] 5. Brandi, C. et al. “The Role of Carbon Dioxide Therapy in the Treatment of 14 Chronic Wounds”. In Vivo. 2010, 24: 223-226.

WE

Claims (57)

CLAIM :

1. Use of a dry powder composition in the manufacture of a medicament for desloughing, debriding, wound treating, or exfoliating applications, the composition comprising: - honey, wherein the honey is in a dry granular form; - at least one pharmaceutically acceptable bicarbonate (HCO ) salt in a dry granular form; and - at least one organic acid in dry granular form, wherein the at least one organic acid is additional to organic acids naturally occurring in honey; wherein the medicament is formulated for topical administration, and provided that the use is not cosmetic.

2. The use according to claim 1, wherein said honey is produced by bees that have fed exclusively on Leptospermum plant species.

3. The use according to claim 1 or 2 wherein said honey is Manuka honey.

4. The use according to claim 3 wherein said Manuka honey has a pH 3.2 to 4.5 and a minimum methylglyoxal concentration of 150 mg/kg.

5. The use according to any one of claims 1 to 4, wherein said at least one pharmaceutically acceptable bicarbonate salt is sodium bicarbonate (NaHCO ), potassium bicarbonate (KHCO ), ammonium bicarbonate ((NH )HCO ) or any combination thereof. 3 4 3

6. The use according to claim 5, wherein said bicarbonate salt is sodium bicarbonate (NaHCO ).

7. The use according to any one of claims 1 to 6, wherein the at least one bicarbonate salt is present at 2.5 to 20% w/w of said composition.

8. The use according any one of claims 1 to 6, wherein the at least one bicarbonate salt is present at 5 to 15% w/w of said composition.

9. The use according to any one of claims 1 to 6, wherein the at least one bicarbonate salt is present at 7.5 to 12.5% w/w of said composition.

10. The use according to any one of claims 1 to 9, wherein the pH of the composition is adjustable from a pH of about 5 to a pH of about 7.4.

11. The use according to any one of claims 1 to 10, wherein the at least one organic acid is additional to organic acids naturally occurring in honey.

12. The use according to any one of claims 1 to 11, wherein the at least one organic acid is gluconic acid, lactic acid, acetic acid, butyric acid, citric acid, malic acid, pyroglutamic acid, succinic acid, ascorbic acid, glycolic acid, mandelic acid, or any combination thereof.

13. The use according to any one of claims 1 to 11, wherein the at least one organic acid is ascorbic acid.

14. The use according to any one of claims 1 to 13, wherein the at least one organic acid is present at a concentration of 0.2 to 10% w/w of said composition.

15. The use according to any one of claims 1 to 13, wherein the at least one organic acid is present at a concentration of 1 to 5% w/w of said composition.

16. The use according to any one of claims 1 to 15, further comprising at least one thickening agent.

17. The use according to claim 16, wherein the at least one thickening agent is a polysaccharide, a starch, agar, alginic acid or a salt thereof, carrageenan, pectin, beta glucan, xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, or any combination thereof.

18. The use according to claim 16 or 17, wherein said at least one thickening agent is xanthan gum.

19. The use according to any one of claims 16 to 18, wherein said at least one thickening agent is present at a concentration of 0.2 to 6% w/w.

20. The use according to any one of claims 1 to 19, further comprising at least one surfactant.

21. The use according to claim 20, wherein the surfactant is sodium lauryl sulfate.

22. The use according to any one of claims 1 to 21, further comprising at least one proteolytic enzyme, wherein the at least one proteolytic enzyme is papain, actinidin, bromelain, ficain, cucumisin, or a bacteria-derived proteolytic enzyme.

23. The use according to claim 22, wherein said proteolytic enzyme is actinidin.

24. The use according to claim 23, wherein said actinidin is provided in kiwi fruit or an extract thereof.

25. The use according to any one of claims 1 to 24, further comprising at least one preservative, wherein the preservative is methyl paraben, propyl paraben, butyl paraben, imidazolidinyl urea, tetradecyl-trimethyl ammonium bromide (Cetrimide®), sodium benzoate, potassium sorbate, thymol or polyaminopropyl biguanide.

26. The use according to any one of claims 1 to 25, wherein the medicament is formulated for application using a carrier or applicator.

27. The use according to any one of claims 1 to 26, wherein the medicament further comprises a pharmaceutically acceptable carbonate (CO ) salt.

28. The use according to claim 27, wherein the carbonate salt is sodium carbonate.

29. A cosmetic method of desloughing, debriding or exfoliating a region of skin of a mammal, the method comprising applying to the region a topical composition comprising: - honey in dry granular form; - at least one pharmaceutically acceptable bicarbonate (HCO ) salt in dry granular form; and, - at least one organic acid in dry granular form, wherein the at least one organic acid is additional to organic acids naturally occurring in honey.

30. The cosmetic method according to claim 29, wherein said honey is produced by bees that have fed exclusively on Leptospermum plant species.

31. The cosmetic method according to claim 29 or 30, wherein said honey is Manuka honey.

32. The cosmetic method according to claim 31 wherein said Manuka honey has a pH 3.2 to 4.5 and a minimum methylglyoxal concentration of 150 mg/kg.

33. The cosmetic method according to any one of claims 29 to 32, wherein the at least one pharmaceutically acceptable bicarbonate salt is sodium bicarbonate (NaHCO ), potassium bicarbonate (KHCO ), ammonium bicarbonate ((NH )HCO ) or any combination 3 4 3 thereof.

34. The cosmetic method according to claim 33, wherein said bicarbonate salt is sodium bicarbonate (NaHCO ).

35. The cosmetic method according to any one of claims 39 to 34, wherein the at least one bicarbonate salt is present at 2.5 to 20% w/w of said composition.

36. The cosmetic method according any one of claims 29 to 34, wherein the at least one bicarbonate salt is present at 5 to 15% w/w of said composition.

37. The cosmetic method according to any one of claims 29 to 34, wherein the at least one bicarbonate salt is present at 7.5 to 12.5% w/w of said composition.

38. The cosmetic method according to any one of claims 29 to 37, wherein the pH of the composition is adjustable from a pH of about 5 to a pH of about 7.4.

39. The cosmetic method according to any one of claims 29 to 38, wherein the at least one organic acid is additional to organic acids naturally occurring in honey.

40. The cosmetic method according to any one of claims 29 to 39, wherein the at least one organic acid is gluconic acid, lactic acid, acetic acid, butyric acid, citric acid, malic acid, pyroglutamic acid, succinic acid, ascorbic acid, glycolic acid, mandelic acid, or any combination thereof.

41. The cosmetic method according to any one of claims 29 to 39, wherein the at least one organic acid is ascorbic acid.

42. The cosmetic method according to any one of claims 29 to 41, wherein the at least one organic acid is present at a concentration of 0.2 to 10% w/w of said composition.

43. The cosmetic method according to any one of claims 29 to 41, wherein the at least one organic acid is present at a concentration of 1 to 5% w/w of said composition.

44. The cosmetic method according to any one of claims 29 to 43, further comprising at least one thickening agent.

45. The cosmetic method according to claim 44, wherein the at least one thickening agent is a polysaccharide, a starch, agar, alginic acid or a salt thereof, carrageenan, pectin, beta glucan, xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, or any combination thereof.

46. The cosmetic method according to claim 44 or 45, wherein said at least one thickening agent is xanthan gum.

47. The cosmetic method according to any one of claims 44 to 46, wherein said at least one thickening agent is present at a concentration of 0.2 to 6% w/w.

48. The cosmetic method according to any one of claims 29 to 47, further comprising at least one surfactant.

49. The cosmetic method according to claim 48, wherein the surfactant is sodium lauryl sulfate.

50. The cosmetic method according to any one of claims 29 to 49, further comprising at least one proteolytic enzyme, wherein the at least one proteolytic enzyme is papain, actinidin, bromelain, ficain, cucumisin, or a bacteria-derived proteolytic enzyme.

51. The cosmetic method according to claim 50, wherein said proteolytic enzyme is actinidin.

52. The cosmetic method according to claim 51, wherein said actinidin is provided in kiwi fruit or an extract thereof.

53. The cosmetic method according to any one of claims 29 to 52, further comprising at least one preservative, wherein the preservative is methyl paraben, propyl paraben, butyl paraben, imidazolidinyl urea, tetradecyl-trimethyl ammonium bromide (Cetrimide®), sodium benzoate, potassium sorbate, thymol or polyaminopropyl biguanide.

54. The cosmetic method according to any one of claims 29 to 53, wherein the medicament is formulated for application using a carrier or applicator.

55. The cosmetic method according to any one of claims 29 to 54, wherein the medicament further comprises a pharmaceutically acceptable carbonate (CO ) salt.

56. The cosmetic method according to claim 55, wherein the carbonate salt is sodium carbonate.

57. The use of claim 1, substantially as herein described with reference to any one of the Examples and/or