US20170360982A1

US20170360982A1 - Wound Healing Composition Involving Mineral Ions and Methylglyoxal, and Methods of Use

Info

Publication number: US20170360982A1
Application number: US15/637,845
Authority: US
Inventors: Mark Richard Wardell; Kenneth A. Sabacinski
Original assignee: Pelogenix LLC
Current assignee: Pelogenix LLC
Priority date: 2015-01-29
Filing date: 2017-06-29
Publication date: 2017-12-21
Also published as: US20170216369A1; US20200054687A1; WO2016123539A1; US10695382B2; CA3009754A1; CA3009754C; US20160220722A1; US10500235B2

Abstract

The invention provides compositions based on an effective amount of an active anti-inflammatory ingredient of mineral solids fortified with methylglyoxal antibacterial activity for the treatment of wounds; and methods of treating a wound, comprising contacting a wound with the above compositions or a wound dressing containing the above compositions.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is a divisional of and claims priority to currently pending U.S. Nonprovisional application Ser. No. 15/010,896, entitled “Wound Healing Compositions Involving Medicinal Honey, Metal Ions, and Methylglyoxal, and Methods of Use,” filed Jan. 29, 2016, which claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 62/109,369, entitled “Wound Healing Compositions Involving Medicinal Honey, Metal Ions, and Methylglyoxal, and Methods of Use,” filed Jan. 29, 2015, the contents of each of which are hereby incorporated by reference into this disclosure.

FIELD OF THE INVENTION

The invention broadly relates to a wound healing composition, more specifically to a wound healing composition including various combinations of inorganic minerals, salts, methylglyoxal, and pharmaceutically acceptable carriers.

BACKGROUND OF THE INVENTION

Undesirable and dangerous side effects and adverse drug interactions are well known for the predominantly synthetic organic pharmaceuticals that have been widely administered over the past several decades. These adverse effects have led many research groups to go back and study, in greater detail, the medicinal properties and mechanisms of action of many natural compounds. Ancient cultures have long been aware of the medicinal properties of natural products, such as compounds derived from botanical sources, and compounds from the seas. The subject matter of the present invention involves novel medicinal activities associated with natural products.
In one embodiment of the present invention, various mineral ions, originally derived from both botanical and oceanic origins and identified to have various medicinal activities are combined into a therapeutic composition.
Plants have been used for medicinal purposes since before recorded history. Ancient Chinese and Egyptian papyrus writings describe medicinal uses for plants as early as 3,000 BC. While some cultures, such as Africans and Native Americans, have used botanical sources in their healing remedies, other cultures including the Chinese and Indians have developed medicinal systems, such as Traditional Chinese Medicine and Ayurveda, respectively, in which botanicals were used. Modern researchers have found that people in different parts of the world used the same or similar plants for the same purposes.
In the early 19^thCentury, when chemical analysis first became available, scientists began to extract and modify the active ingredients from botanical sources. Today almost one quarter of pharmaceutical drugs are derived from botanicals. Recently, the World Health Organization (WHO) estimated that 80% of people worldwide rely on botanical medicines for some part of their primary health care. Botanical medicine is being taught more in medical schools and pharmacy schools, and more health care providers are learning about the positive effects of using botanical medicine to help treat health conditions. In Germany, for example, about 600-700 plant-based medicines are available and are prescribed by some 70% of German physicians. In the past 20 years in the United States, public dissatisfaction with the cost of prescription medications and their extensive adverse effects, combined with an interest in returning to more natural remedies, has led to an increase in botanical medicine use. Botanical medicine is used to treat many conditions, such as asthma, eczema, premenstrual syndrome, menopausal symptoms, rheumatoid arthritis, migraine, chronic fatigue, irritable bowel syndrome, cancer and chronic wounds among others. Today, nearly one-third of Americans use herbs and one study found that 90% of arthritic patients use alternative therapies such as botanical medicine.
One such botanical remedy originated in American Indian folklore as a treatment for hard-to-heal wounds. It was handed down to the pioneers in South Carolina as an extract of ash derived specifically from Red Oak Bark (Quercus rubra) grown in the region around Piedmont, S.C. A paste of this extract that was applied to a hard-to-heal wound or skin ulcer proved to be very effective at healing the wound. It is now known that Red Oak Bark contains specific storage cells that collect and concentrate certain mineral ions from the soil in which they grow, and that the soil around Piedmont, S.C. contains a high level of specific minerals that are key for healing wounds. Chemical analyses of the extracts from the ash of Red Oak Bark led to the identification of the minerals responsible for the wound healing effect and a safe and efficacious version of the active minerals, which included potassium, calcium, zinc and rubidium is now manufactured by combining these minerals in a proprietary formulation and used for chronic wound therapy. U.S. Pat. No. 5,080,900 described the oak bark ash extract and U.S. Pat. Nos. 6,149,947 and 7,014,870 described the synthetic version of the formulation. All three patents are now expired.
In one study, Weindorf et al., (2012) demonstrated that the synthetic formulation of these ions prepared in the pharmaceutically-accepted carrier of polyethylene glycols used to treat over 300 therapy-refractory wounds, demonstrated a wound size reduction of at least 50% in 73% of the patients, where a wound size reduction of at least 50% is predictive of successful wound closure. It is thought that the wound-healing activity of the specific mixture of potassium, calcium, zinc, and rubidium in this study is anti-inflammatory by down-regulating the expression of proteases in wounds and reducing reactive oxygen species.
As with plants, the marine ecosystem has been a source of therapeutics for the treatment of human diseases. In recent times, cancer drugs have been developed from marine sources. Cytosar, a staple treatment for leukemia and lymphoma was derived from a Caribbean sea sponge. Other anti-cancer drugs have been derived from tunicates, and a potent medicine for the treatment of chronic pain, more powerful than morphine, has been derived from the venom of cone snails that inhabit the reefs of Australia, Indonesia and the Philippines. The waters of the Dead Sea have been renowned for their therapeutic effects since ancient times. Galenus, a prominent first Century Greek physician, stated that salt water from the Dead Sea was good for the treatment of arthritis, eczema, muscular pain, rheumatism, and psoriasis, and the Jewish-Roman historian, Flavius Josephus, wrote two thousand years ago that the salts from the Dead Sea heal the human body and are therefore used in many medicines.
The water of the Dead Sea is unique compared to other seas and lakes, in its high concentration of salts—Dead Sea water contains 330 g of minerals per liter (33%). The salt concentration is between 7-10 times that of the oceans, which typically contains 3.5% minerals. The mineral composition of the Dead Sea is also significantly different from that of ocean water. Whereas the major salt constituent of ordinary seawater is sodium chloride (NaCl), Dead Sea salt is rich in MgCl₂, CaCl₂, KCl, MgBr₂, and CaSO₄. The concentration of ionic species present in the Dead Sea water is: magnesium (40.65 g/L), sodium (39.15 g/L), calcium (16.86 g/L), potassium (7.26 g/L), chloride (212.4 g/L), bromide (5.12 g/L), sulfate (0.47 g/L), and bicarbonate (0.22 g/L). The bromide ion concentration is the highest of all waters on the earth and serum bromide levels have been shown to increase up to 4-fold after bathing in the Dead Sea for four weeks, as a result of entering the circulation and internal organs through the skin. Metal ions are required for many critical functions in humans. Scarcity of some metal ions often leads to disease. Four main group metals (Na, K, Mg, and Ca) and 10 transition metals (V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Mo, and Cd) are currently known, or thought, to be required for normal biological function. It is believed that the therapeutic properties of the Dead Sea are due to a large extent to the presence of magnesium, potassium and bromide.
Various cultures and groups of people have visited the Dead Sea for therapy, dating back to the time of the ancient Egyptians, utilizing the salt in various unguents and skin creams, as well as soaps, just as it is used today. The Dead Sea has taken on a new dimension today: modern science has proven the therapeutic and rejuvenating properties of its unique mineral content. The Dead Sea has become a renowned center for natural health with people coming from around the world to bathe in mineral-rich waters. Balneotherapy emerged as an important treatment modality in the 1800s, first in Europe and then in the United States. After seeing decline in use for almost 50 years, balneotherapy in the Dead Sea has experienced resurgence in popularity over the past three decades, at the same time as a new recognition of the safety and efficacy of natural remedies has fueled a resurgence in popularity for these treatment modalities. The major dermatological diseases that are frequently treated by balneotherapy in the Dead Sea with a high rate of success are psoriasis and atopic dermatitis. Both magnesium and potassium ions in Dead Sea water have a specific inhibitory capacity on the uncontrolled proliferation of psoriatic dermis grown in tissue culture.
Magnesium salts, the prevalent minerals in Dead Sea salt, are known to exhibit favorable effects in inflammatory diseases. In one study of atopic dry skin, bathing in magnesium-rich Dead Sea salt solution improved skin barrier function, enhanced skin hydration, and reduced inflammation. In other in vitro and in vivo studies, magnesium ions inhibit the number and function of epidermal Langerhans cells that contribute to inflammatory skin diseases by presenting alloantigens to T lymphocytes, thereby activating these cells to release pro-inflammatory cytokines. In one study, the reduced antigen presenting activity of MgCl₂-treated Langerhans cells was associated with suppression of constitutive tumor necrosis factor (TNF)-α production by the epidermal cells in vitro. Another study showed that Dead Sea water inhibited the production and/or release of the pro-inflammatory cytokines, interleukin (IL)-2 and interferon (INF)-γ, from Th1 lymphocytes. Others confirmed down-regulation of the pro-inflammatory cytokines TNF-α and IL-1, and an up-regulation of insulin-like growth factor (IGF)-1 following balneotherapy in the Dead Sea.
Magnesium impacts more than 325 enzyme systems in the human body. For example, it is a rate-limiting factor in the activation of epidermal adenylate cyclase and consequently in the production of cyclic adenosine monophosphate (cAMP). A decrease of cAMP and concomitant increase in cyclic guanosine monophosphate (cGMP) has been implicated in excessive cellular proliferation, a major element of the psoriatic state. Balneotherapy in Dead Sea water has also been applied to the treatment of various inflammatory rheumatic diseases such as rheumatoid arthritis and psoriatic arthritis. In a study of Dead Sea water in the treatment of patients with rheumatoid arthritis, Dead Sea water produced statistically significant clinical improvements in most parameters for up to one month following cessation of treatment, whereas treatment with sodium chloride water did not. Dead Sea balneotherapy for 14 days also produced significant clinical improvements in knee osteoarthritis, which lasted for at least 1 month following cessation of treatment. Many other diseases are also treated by balneotherapy in the Dead Sea, including chronic ulcers.
Hypertonic Dead Sea water has also been shown to be efficacious in the treatment of allergic rhinitis. The two common treatments for such sinonasal disease are intranasal rinsing with normal saline solution or corticosteroid rinses, and both have positive effects on the physiology of nasal mucosa. In a study comparing Dead Sea saline spray with NaCl solution, the hypertonic Dead Sea saline solution proved efficacious in mild-to-moderate allergic rhinitis, including improving mucociliary clearance, whereas no significant improvement was seen with the nasal saline spray.
DePootere et al., (2011) demonstrated that the addition of magnesium and bromide to some of the botanically-derived inorganic metal ions gave an enhanced anti-inflammatory and wound healing effect in an animal model of chronic rhinosinusitis. The present invention adds antibacterial and anti-biofilm activities to the anti-inflammatory activity of the mineral salts composed of a mixture of active mineral ion ingredients from Red Oak Bark, and the Dead Sea.
Antimicrobial resistance is an increasing clinical problem precipitated by the inappropriate use of antibiotics in the latter parts of the 20^thCentury. This problem, coupled with the lack of novel therapeutics in the development pipeline, means antimicrobial resistance is reaching a crisis point, with an expected annual death rate of ten million people worldwide by 2050. To reduce, and to potentially remedy this problem, many researchers are looking into natural compounds with antimicrobial and/or anti-virulence activity.
The natural non-peroxide antibacterial compound, methylglyoxal, found in honey derived from the nectar of plants from the Leptospermum genus of shrubs and small trees of the myrtle family, commonly known as tea trees, has bactericidal activities against antibiotic-sensitive and antibiotic-resistant strains of both gram-positive and gram negative bacteria in planktonic form and biofilm colonies. In the present invention we teach that MGO can be added to the mineral ions derived from Red Oak Bark and the Dead Sea to supplement the anti-inflammatory and antioxidant activities of these mineral ions with antibacterial activity.
The antibacterial effect of MGO was largely elucidated after observing that honey derived from the Manuka Tea Tree (Leptospermum scoparium) in New Zealand killed bacteria when applied to infected wounds. During these studies, manuka honey was found to retain most of its antibacterial activity when catalase was added to the honey to knock out its peroxide-mediated antibacterial activity. The non-peroxide activity of Manuka honey was found to be due to methylglyoxal (originally referred to as UMF [Unique Manuka Factor]). MGO is generated in Leptospermum honey largely by non-enzymatic Maillard reactions from dihydroxyacetone present in the nectar of Leptospermum flowers, and is found in highest concentrations in L. scoparium and L. polygalifolium, the Australian Jelly Bush.
In the 1970s methylglyoxal was found to be produced from dihydroxyacetone phosphate in Escherichia coli (E. coli), initiating a bypass of the glycolytic pathway. It was suggested that MGO inhibits protein synthesis by reacting with guanine residues in RNA and its precursors. It also inhibits DNA synthesis by reacting with guanine residues in DNA and its precursors. More recently, the mechanism of the antibacterial action of MGO has mainly been elucidated against two prominent wound pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. While S. aureus is a gram-positive bacterium and P. aeruginosa is a gram-negative bacterium, it is perhaps not surprising that the mechanisms of antibacterial action by MGO differs for the two organisms.
MGO causes cell disruption to the regular cell division process of S. aureus. Under optimal conditions, bacterial cells duplicate and segregate their chromosome, forming a proteinaceous ring (the septum) across the midcell, creating two daughter cells that are still joined together. The completion of cell division occurs when peptidoglycan (murein) hydrolases degrade the cell wall between the two daughter cells, allowing separation. MGO inhibits the activity of murein hydrolase, causing a build-up of septated non-dividing cells.
In contrast to the mechanism observed in the gram-positive S. aureus, MGO causes the loss of cellular integrity in the gram-negative P. aeruginosa, leading to extensive cell lysis and cell death. P. aeruginosa modulates its structural integrity through the production of a key anchor protein, outer membrane protein F (OprF). This protein provides a vital link between the outer membrane and an underlying peptidoglycan layer, ensuring cell envelope homeostasis and regular cell shape. Reduced OprF expression has been observed in populations treated with MGO, and a concomitant increase in membrane blebbing and cell lysis has also been detected.
In other studies with P. aeruginosa, MGO has been shown to suppress the class I master regulators (FleQ and FliA), inhibiting the regulatory cascade required for flagellum production resulting in a significant reduction in invasive virulence. Resistance to bactericidal agents is often associated with multidrug efflux systems, which decrease cellular drug accumulation. In gram-negative bacteria, systems belonging to the resistance/nodulation/division (RND) family are particularly effective in generating resistance because they form a tripartite complex with the periplasmic proteins of the membrane fusion protein family and an outer membrane channel, ensuring that drugs are pumped out directly to the external medium.
P. aeruginosa expresses several RND-type multidrug efflux systems, including MexAB-OprM, MexCD-OprJ, MexEF-OprN, MexHI-OpmD, and MexXY, which are significant determinants of multidrug resistance in laboratory and clinical isolates. MGO is not recognized by multidrug systems in P. aeruginosa and is therefore not exported from the cell. MGO equally inhibits drug-susceptible P. aeruginosa and MDRP at concentrations of 128-512 μg/mL (1.7-7.1 mM). MGO also effectively kills Escherichia coli, and Salmonella enterica.
MGO is also biofilm-cidal. Manuka honey was found to inhibit biofilm formation and to remove preformed biofilms. Biofilm formation of colonies of five clinical isolates of each of S. aureus, P. aeruginosa, Klebsiella spp. and Proteus mirabilis, isolated from diabetic foot ulcers were inhibited by manuka honey, and established biofilm colonies were killed by manuka honey (Abbas, 2014). This finding has been traced to the predominant bactericidal agent in manuka honey, MGO, although other components in manuka honey contribute. Jervis-Bardy et al., (2011) demonstrated that MGO-only solutions were biofilm-cidal to five S. aureus strains (four clinical isolates and one reference strain). It has been speculated that the biofilm-cidal activity of MGO may originate from its quorum sensing inhibiting activity. Quorum sensing controls biofilm formation.
Another important modern-day gram-positive, anaerobic nosocomial pathogenic superbug is Clostridium difficile (C. difficile), which currently accounts for 30-50% of hospital-acquired infections. MGO inhibited the growth of three strains of C. difficile one of which was a laboratory strain and two were derived from clinical strains all with MIC values of 6.25%, similar to that for P. aeruginosa (5.5-8.7%). Furthermore, MGO was found to be bactericidal for C. difficile as determined by its Minimal Bactericidal Concentration/Minimal Inhibitory Concentration (MBC/MIC) ratio of 1, where MBC/MIC ratios less than or equal to 4 indicate bactericidal activity and MBC/MIC ratios greater than or equal to 16 indicate bacteriostatic activity. MGO is therefore a good bactericidal agent against wound pathogens and was used in this invention to add bactericidal activity to the anti-inflammatory and antioxidant activities of a mineral ion composition, to yield a single effective therapeutic composition of natural antagonists against the two most important factors that prevent wound healing; chronic inflammation (caused by elevated proteases) and infection (caused by wound bacteria).
As can be derived from the variety of devices and methods directed at wound healing compositions, many strategies have been contemplated to accomplish the desired end. Heretofore, widely administered synthetic organic pharmaceuticals are commonly associated with undesirable side effects and adverse drug interactions. Thus, there is a long-felt need for more natural wound healing compositions. There is a further long-felt need for wound healing compositions involving, mineral ions and methylglyoxal, and their corresponding methods of use.

BRIEF SUMMARY OF THE INVENTION

The composition of the present invention includes a mixture of ingredients originally isolated from a botanical source, with ingredients originally isolated from the Dead Sea, to yield an active source of anti-inflammatory activity that down-regulates protease gene expression in chronic wounds. To this embodiment of this inventive composition of a mixture of botanical- and oceanic-derived inorganic minerals, methylglyoxal is added to supplement its anti-inflammatory activity with non-peroxide antibacterial activity. This composition, herein referred to as ‘MVE ointment with MGO,’ (wherein ‘MVE’ denotes ‘multivalent electrolytes’) includes an active anti-inflammatory ingredient of inorganic mineral solids including salts of magnesium, potassium, calcium, zinc, rubidium, bromide, and sulfate to which is added non-peroxide antibacterial activity in the form of the natural compound, methylglyoxal.
The inventive composition is used in aqueous, ointment, or wound-dressing formulations to modulate biochemical mechanisms associated with wound healing, including decreasing both wound protease activities and active infection.
These and other objects and advantages of the present invention will be readily appreciable from the following description of preferred embodiments of the invention and from the accompanying examples and claims.

BRIEF DESCRIPTION OF THE DRAWING

The nature and mode of operation of the present invention will now be more fully described in the following detailed description of the invention taken with the accompanying FIGURE, in which:

FIG. 1,

panels

1A, 1B, 1C, 1D, 1E, and 1F shows treatment using the composition of the present invention, ‘MVE ointment with MGO’ wound dressings of a diabetic foot ulcer.

DETAILED DESCRIPTION OF THE INVENTION

While the present invention is described with respect to what is presently considered to be the preferred aspects, it is to be understood that the invention as claimed is not limited to the disclosed aspects.
Furthermore, it is understood that this invention is not limited to the particular methodology, materials and modifications described and as such may, of course, vary. It is also understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to limit the scope of the present invention, which is limited only by the appended claims.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this invention belongs. Although any methods, devices or materials similar or equivalent to those described herein can be used in the practice or testing of the invention, the preferred methods, devices, and materials are now described.
The present invention provides an exemplary wound healing composition. The composition includes a pharmaceutically-acceptable carrier, an effective amount of an active mixture of inorganic minerals to reduce inflammation at a wound site, and an amount of methylglyoxal (500-2000 mg per kg of final product) that effectively reduces the number of viable microorganisms at a wound site. The active mixture of inorganic minerals include, but are not limited to, a magnesium salt, a potassium salt, a calcium salt, a zinc salt, a rubidium salt, and a bromide salt, wherein each of the salts include a pharmaceutically-acceptable anion including bromide, chloride, citrate, and sulfate.
In any embodiment of the composition, the active mixture of inorganic minerals includes at least one of 1.5-75 parts of magnesium ions, 0.5-75 parts of potassium ions, 0.001-10 parts of calcium ions, 0.0001-10 parts of zinc ions, up to 5 parts of rubidium ions, 10-80 parts of chloride ions, 0-40 parts of citrate ions, 0.001-20 parts of bromide ions, and up to 20 parts of sulfate ions, said parts being expressed as parts by weight of the total inorganic mineral solids. In any of the above embodiments, methylglyoxal is added to a concentration in the final formulation of between 500-2000 mg per kg.
The composition includes a pharmaceutically-acceptable carrier, such as but not limited to a carrier including at least one of water, polyethylene glycol, an ointment, and a cream base, which results in a therapeutic composition having a pH in the range of 3-7.5, inclusive. Preferably, the composition has a pH in the range of 3.5-6.5, inclusive. As used herein, a composition comprising ‘MVE salts’ refers to a composition that includes magnesium, potassium, calcium, zinc, and rubidium cations, together with pharmaceutically-acceptable counterions including bromide, chloride, citrate, and sulfate anions.
In another aspect, the present invention provides a wound dressing. The wound dressing is used with any of the above embodiments of the composition and a support. In any embodiment of the dressing, the support includes, but is not limited to, a fibrous gauze material, a hydrogel, a foam, a film, a hydrocolloid, an alginate, a collagen, or a combination of any two or more of the afore-mentioned.
In yet another embodiment, the present invention includes a method of treating a wound. The method includes contacting a wound with any of the above embodiments of the wound dressing.
A ‘chronic wound,’ ‘non-healing wound,’ ‘slow-to-heal wound,’ or ‘stalled wound,’ as used herein, refers to a wound that fails to heal over a 4-12 week timeframe from inception of the wound to complete closure of the skin at the wound site. Such wounds commonly include external dermal wounds or wounds of mucosal membranes such as sinonasal or endometrial wounds.
Skin wounds designated as ‘chronic’ or ‘non-healing’ or ‘slow-to-heal’ or ‘stalled’ are commonly observed in clinical settings as venous leg ulcers, diabetic foot ulcers, pressure ulcers, arterial ulcers, ulcers of mixed etiology, burns, or non-healing surgical wounds. Other types of non-healing wounds are observed in less frequent conditions, such as, fistulae, dermatitis or vasculitis wounds, skin cancers, and radiation burns. This list is not exhaustive and is provided to show examples of such non-healing wounds. Differentiated from ‘acute’ wounds that spontaneously heal without complications in a matter of days or weeks through the four normal phases of the ‘wound healing curve’ (hemostasis, inflammation, proliferation, and remodeling), chronic wounds may persist for months or years and occasionally can last a lifetime, and are therefore commonly referred to as ‘non-healing’ wounds. There is a need for treatment of any of these types of non-healing wounds since spontaneous healing has failed to occur. In chronic wounds, at the cellular biological level, there is commonly a prolonged inflammatory phase often caused by elevated proteases or active infection.
In yet another embodiment of the present invention, the composition is used prophylactically to prevent surgical incisions in high-risk patients from post-operative wound dehiscence and non-healing. More than 53 million people undergo surgical procedures annually in the United States, with about half of these occurring under general anesthesia. Post-operative wound dehiscence has been investigated in several studies and a small incidence has shown to be an issue of concern in all age groups, including the pediatric population, but with a higher incidence in the older population. The failure of these surgical wounds to heal in a normal time frame pushes them into the category of chronic wounds. Diabetes, obesity, cancer therapy, and vascular insufficiency and other abnormalities, which are all increasing in incidence in the Western population, contribute to delayed healing and are considered risk factors. The prophylactic application of the present inventive composition to surgical incisions post-operatively in ‘high-risk’ patients will aid in reducing the incidence of non-healing surgical wounds.
Sometimes prolonged inflammation due to elevated wound proteases and active infection occur simultaneously and prevent wounds of the skin on mucosal membranes from healing. The present disclosure relates to a composition, carriers, and methods for treating wounds of the skin and the mucosal membranes to counteract these pathological conditions. The composition includes an active ingredient of a mixture of inorganic minerals originally derived from botanical and Dead Sea sources and including magnesium, potassium, calcium, zinc, rubidium, bromide, and sulfate. The components of the inventive composition surprisingly provide a synergistic effect that results in the suppression of the accumulation of a biochemical marker (e.g. proteases, and pro-inflammatory cytokines) associated with inflammation, and the up-regulation of other biochemical markers (e.g. growth factors and protease inhibitors) associated with wound healing. In addition, the components of the inventive composition provides antibacterial activity that acts concomitantly with the anti-inflammatory activity to also provide suppression of active infections.
The composition according to the present invention is useful for treating common chronic wounds, such as venous leg ulcers, diabetic foot ulcers, pressure ulcers, arterial ulcers, burns, non-healing surgical wounds, chronic rhinosinusitis and metritis. In addition, the composition according to the present invention is also useful for treating abrasions, lacerations, minor cuts, scalds and burns, and other partial thickness wounds. The composition that is useful includes, but is not limited to methylglyoxal, magnesium, potassium, calcium, zinc, rubidium, bromide, and sulfate. The composition is advantageously applied in a cream or ointment base that is applied to wounds until they are healed (3-8 months) with wound dressing changes every 24-96 hours. Alternatively, the wound healing composition of the present invention is impregnated into or associated with carrier dressing supports (e.g. fibrous gauze, hydrogel, foam, film, hydrocolloid, collagen, or alginate), which are applied to wounds for the times described above.
The present disclosure further provides a method for treating the wound. In some embodiments, the method includes contacting a wound with the composition of the present disclosures wherein the composition includes a pharmaceutically-acceptable carrier; an effective amount of an active ingredient of inorganic solids comprising a magnesium salt, a potassium salt, a calcium salt, a zinc salt, a rubidium salt, a bromide salt, and a sulfate salt effective to reduce the protease activity at a wound site; and the natural non-peroxide antibacterial compound, methylglyoxal comprising of an amount between 500-2000 mg per kg of finished wound healing composition, effective to reduce the number of viable microorganisms at a wound site. The composition is applied to the wound, for example, in a liquid, (e.g. by irrigating or lavaging the wound with the liquid) or in a gel or an ointment. Liquid compositions provide immediate availability of the ions and methylglyoxal to the healing tissue. In contrast, gels or ointments can provide regulated delivery of the mineral ions, and methylglyoxal to the healing tissue over a sustained regulated period of time. In some embodiments, the composition is applied to a wound dressing, which is subsequently applied to the wound. Advantageously, a dressing including the composition is contacted with the wound until it is healed (3-8 months) with wound dressing changes every 24-96 hours, thereby providing a moist environment enriched with the MVE ions and methylglyoxal to facilitate healing of the skin or mucosal membrane.

Embodiments

Embodiment 1 is a composition including a pharmaceutically-acceptable carrier, an effective amount of an active ingredient of inorganic solids including a magnesium salt, a potassium salt, a calcium salt, a zinc salt, a rubidium salt, a bromide salt, and a sulfate salt; and methylglyoxal as an antimicrobial at an amount between 500-2000 mg per kg, which is effective to reduce the number of viable microorganisms at a wound site, wherein each of the salts includes a pharmaceutically-acceptable counterion.
Embodiment 2 is the composition of embodiment 1, wherein the active ingredient of inorganic solids includes 1.5-75 parts of magnesium ions, 0.5-75 parts of potassium ions, 0.001-10 parts of calcium ions, 0.0001-10 parts of zinc ions, up to 5 parts of rubidium ions, 0.001-20 parts of bromide ions, and up to 20 parts of sulfate ions (said parts being expressed as parts by weight of the total weight of inorganic solids), and methylglyoxal representing 500-2000 mg per kg of the final composition.
Embodiment 3 is a wound dressing including the composition of any of embodiments 1 or 2; and a support.
Embodiment 4 is the wound dressing of embodiment 3, wherein the support includes a fibrous gauze material, a hydrogel, a foam, a film, a hydrocolloid, an alginate, a collagen, or a combination of any two or more of the afore-mentioned.
Embodiment 5 is a method of treating the wound, including contacting a wound with the composition of either embodiments 1 or 2.
Embodiment 6 is a method of treating a wound, including treating a wound with the wound dressing of either one of embodiments 3 or 4.

Examples

Objects and advantages of this invention are further illustrated by the following examples, but the particular materials and amounts thereof recited in these examples, as well as other conditions and details, should not be construed to unduly limit this invention.
Example 1 demonstrates embodiments 1 and 2 of the composition.
As shown in FIG. 1, Panels 1A, 1B, 1C, 1D, 1E, and 1F, show treatment using dressings impregnated with the composition of the present invention, herein referred to as ‘MVE ointment with MGO,’ of a diabetic foot ulcer. ‘MVE ointment with MGO’ was prepared in two parts as follows: Part A, the PEG phase, was prepared by adding 34.205 g polyethylene glycol (PEG)-400 to a Pyrex beaker and heating it to 70° C. While constantly stirring the pre-heated PEG-400, 42.274 g PEG-4000 was added slowly until all the solid PEG-4000 was dissolved into the PEG-400 and the PEG phase was left to cool to 50° C. Part B, the aqueous phase, was prepared by weighing 21.21 g deionized water into a glass beaker and, with constant stirring, dissolving into it 1.681 g magnesium chloride, hexahydrate; 0.387 g potassium chloride; 0.02 g magnesium bromide, hexahydrate; 0.0028 g magnesium sulfate, heptahydrate; 0.0164 g calcium chloride, dihydrate; 0.0025 g zinc chloride; 0.0018 g rubidium chloride; and 0.1 g sodium benzoate. The total amount of inorganic minerals, herein referred to as MVE (for multivalent electrolytes), was 2.1115% in weight.
When all the salts had dissolved, the aqueous phase was slowly added to the PEG phase with constant stirring, while the temperature of the PEG phase was initially at 50° C. During the addition of the aqueous phase to the PEG phase, outgassing occurred and the temperature of the solution decreased. The pH of this ointment without adjustment is 4.30 (10-fold dilution of a small aliquot of the ointment in water was used to measure the pH) and the s.g.=1.1 g/mL. The pH of this ointment can be lowered, if desired, by the addition of 50% hydrochloric acid. After the mixture had cooled to 40° C., 0.1 g methylglyoxal was added and the total 100 g of ointment was then transferred to tubes, which were sterilized with gamma radiation and verified as sterile before use. Alternatively, the ointment was prepared and impregnated into acetate non-woven medical grade dressing (approximately 4 g ‘MVE ointment with MGO’ in each 4-inch×5-inch dressing). The dressings are protected with polyethylene liners applied to both sides, and dressings of 4-inch×5-inch are sealed individually in foil pouches constructed of white polyester film fused to aluminum foil that constitutes an excellent barrier. The dressings are then sterilized with gamma radiation and verified as sterile before use.
When applied daily to a diabetic foot ulcer that had become infected and was not healing by previous treatment modalities, the ‘MVE ointment with MGO’ described herein closed the wound in 19 weeks (see Panels 1A-1F). This wound was on the sole of the left foot of an 82-year-old Caucasian woman with hypertension and a 12-year history of type II diabetes. At age 72 she developed neuropathy and at age 80 she developed a diabetic foot ulcer on the sole of her left foot (Panel 1A). The ulcer had previously been treated unsuccessfully with Regranex for 18 months. Subsequently the left foot ulcer was treated with sterile ‘MVE ointment with MGO’ applied to sterile gauze and then to the wound. The ‘MVE ointment with MGO’ dressings applied to the left foot ulcer were changed every 24 hours (the patient was instructed on how to change her own dressings) and the healing progression is depicted in Panels 1A-1F. Panel 1A represents the wound on the day ‘MVE ointment with MGO’ treatment started; Panel 1B, after treatment for 2.5 weeks with ‘MVE ointment with MGO’; Panel 1C, after treatment for 7.5 weeks with ‘MVE ointment with MGO’; Panel 1D, after treatment for 10 weeks with ‘MVE ointment with MGO’; Panel 1E, after treatment for 13 weeks with ‘MVE ointment with MGO’; Panel 1F, left foot DFU healed after treatment for 19 weeks with ‘MVE ointment with MGO’.
Example 2 demonstrates embodiments 1 and 2 of the composition, variant on Example 1.
A composition, herein referred to as ‘PeloSinus,’ was prepared by dissolving 1.645 g magnesium chloride, hexahydrate; 0.548 g potassium citrate, monohydrate; 0.019 g magnesium bromide, hexahydrate; 0.0026 g magnesium sulfate, heptahydrate; 0.017 g calcium chloride, dihydrate; 0.0001 g zinc chloride; 0.002 g rubidium chloride; 0.05 g methylglyoxal; and 0.000065 g citric acid in deionized water to a final volume of 100 mL. The total amount of mineral salts, herein referred to as MVE (for multivalent electrolytes), was 2.2337% (w/v). This solution is isotonic with body fluids, having a tonicity of 0.909 g sodium chloride equivalents per 100 mL, and is used for the treatment of internal mucous membranes such as the sinonasal and endometrial linings. It is also used as a wound wash or cleanser. The pH of the solution thus formulated is 6.5. Prior to use the solution is sterilized, for example by filtration or irradiation.
The anti-biofilm activity of this solution was tested in a well-characterized biofilm assay (published by Desrosiers et al., 2007) and the results are shown in Table 1. Biofilm colonies were grown using two species of bacteria that commonly form biofilm colonies in human wounds, Pseudomonas aeruginosa and Staphylococcus aureus. As can be seen in Table 1, treatment with PeloSinus' solution reduced Pseudomonas aeruginosa biofilm colony formation by more than 1 log and it reduced Staphylococcus aureus biofilm colony formation by more than 4 logs. Both of these are very significant reductions in the abilities of common wound pathogens to establish in a wound environment. Biofilm is present in about 60% of chronic wounds and is very difficult to eliminate.

TABLE 1

Anti-Biofilm Activity of ‘PeloSinus’ Formulation Against
Pseudomonas aeruginosa and Staphylococcus aureus Biofilm Colonies

Saline Control	Saline Control
Pseudomonas aeruginosa	Staphylococcus aureus

	Plate Count	Cells/cm²		Plate Count	Cells/cm²

Drop 1	15	8.00 × 10⁸	Drop 1	5	2.67 × 10⁶
Drop 2	26	1.39 × 10⁹	Drop 2	4	2.13 × 10⁶
Drop 3	24	1.28 × 10⁹	Drop 3	3	1.60 × 10⁶
Drop 4	21	1.12 × 10⁹	Drop 4	4	2.13 × 10⁶
Drop 5	29	1.55 × 10⁹	Drop 5	2	1.07 × 10⁶
Average		1.23 × 10⁹	Average		1.92 × 10⁶
Log		9.09	Log		6.28

‘PeloSinus’	‘PeloSinus’
Pseudomonas aeruginosa	Staphylococcus aureus

	Plate Count	Cells/cm²		Plate Count	Cells/cm²

Drop 1	12	6.40 × 10⁷	Drop 1	4	2.13 × 10²
Drop 2	16	8.53 × 10⁷	Drop 2	3	1.60 × 10²
Drop 3	15	8.00 × 10⁷	Drop 3	2	1.07 × 10²
Drop 4	14	7.47 × 10⁷	Drop 4	2	1.07 × 10²
Drop 5	10	5.33 × 10⁷	Drop 5	5	2.67 × 10²
Average		7.15 × 10⁷	Average		1.71 × 10²
Log		7.85	Log		2.23

Treatment with ‘PeloSinus’	Treatment with ‘PeloSinus’
Anti-Biofilm Results in a 1.24	Anti-Biofilm Results in a 4.05
Log Reduction of	Log Reduction of
Pseudomonas aeruginosa	Staphylococcus aureus
Biofilm (9.09-7.85)	Biofilm (6.28-2.23)

Irrigation of the sinonasal cavities with sterile isotonic ‘PeloSinus’ also demonstrated efficacy in an animal model of chronic rhinosinusitis. Relative to the standard of care, normal saline solution, ‘PeloSinus’ irrigation of the sinuses led to a significantly increased lumen space and faster replenishment of mucocilial cells after surgical mucosal stripping.
Example 3 demonstrates embodiments 1 and 2 of the composition, variant of Examples 1 and 2.
Two compositions (‘PeloWash’ and ‘PeloFusion’) were prepared for the treatment of metritis in thoroughbred, quarter horse and other valuable mares (also for the treatment of metritis in other animals including dogs). These compositions are referred to herein as ‘PeloWash’ and ‘PeloFusion,’ which together with ‘PeloPurge’ (PeloWash+dimethyl sulfoxide [DMSO]), constitute a treatment system, herein referred to as ‘PeloMetriSystem.’ After successful racing and/or performance careers, thoroughbred, quarter horse and other elite performance mares are valuable for breeding. Many such mares, however, either do not get pregnant or can only have one foal, and then contract metritis and can't get pregnant again. Metritis is a disorder characterized by inflammation of the endometrial lining of the uterus and infection with various bacteria, including Klebsiella, Streptococcus, and Pseudomonas spp. The current standard of care uses irrigation of the endometrial lining of the uterus in mare and dogs with saline (or Lactated Ringer's solution) followed by insertion into the uterus of a solution containing antibiotics. Often systemic antibiotics are given simultaneously. But the continuing problem of bacteria developing resistance to antibiotics continues, and many times antibiotic treatment of metritis is either ineffective or causes undesirable reactions in mares. ‘PeloMetriSystem’ offers an alternative treatment and consists of three solutions as follows:
Solution 1, a composition herein referred to as ‘PeloWash,’ is prepared by dissolving 16.872 g magnesium chloride, hexahydrate; 5.628 g potassium citrate, monohydrate; 0.190 g magnesium bromide, hexahydrate; 0.026 g magnesium sulfate, heptahydrate; 0.17 g calcium chloride, dehydrate; 0.001 g zinc chloride; 0.02 g rubidium chloride; and 0.00065 g citric acid in deionized water to a final volume of 1000 mL. This solution is isotonic and has a pH of 6.5.
Solution 2, herein referred to as ‘PeloPurge’ is constituted by mixing 9 parts (by volume) of Solution 1 (‘PeloWash’) with 1 part (by volume) of 90% dimethylsulfoxide (DMSO), just before use.
Solution 3, a composition herein referred to as ‘PeloFusion,’ is prepared by dissolving 16.023 g magnesium chloride, hexahydrate; 5.34 g potassium citrate, monohydrate; 0.190 g magnesium bromide, hexahydrate; 0.026 g magnesium sulfate, heptahydrate; 0.17 g calcium chloride, dehydrate; 0.001 g zinc chloride; 0.02 g rubidium chloride; and 0.00065 g citric acid; 1 g methylglyoxal; and 30 g polyethylene glycol 4000 in deionized water to a final volume of 1000 mL. This solution is isotonic and has a pH of 6.5.
Four mares were treated with this ‘PeloMetriSystem’ as follows: First, each mare was tested for the presence of metritis-causing bacteria in their uterus. The endometrial lining of the uterus was washed with two or three liters of ‘PeloWash.’ This was done by placing 1 L of ‘PeloWash’ at a time into a mare's uterus and relatively quickly withdrawing the solution again. Whether 2 or 3 L was used was determined by how much debris was present in the eluent of each wash. If testing indicated that metritis-causing bacteria was present in a mare's uterus, that mare was next washed with 1 L ‘PeloPurge’ solution, which again was relatively quickly withdrawn after application. Finally, 250 mL of ‘PeloFusion’ solution was placed in a mare's uterus and left there. This solution dissipates naturally—some being absorbed, and some pushed out of the uterus. No systemic antibiotics were used.
Four other ‘control’ mares were treated with the standard of care as follows: First, each mare was tested for the presence of metritis-causing bacteria in their uterus. The endometrial lining of the uterus was washed with two to three liters of normal saline or isotonic Lactated Ringer's solution, 1 L at a time, each liter being relatively quickly withdrawn after insertion. Whether 2 or 3 L was used was determined by how much debris was present in the eluent of each wash. If testing indicated that metritis-causing bacteria was present in a mare's uterus, that mare was next washed with 1 L of 10% DMSO in normal saline or Lactated Ringer's solution, which again was relatively quickly withdrawn after application. Finally, 250 mL of an antibiotic (Amikacin or Timentin) solution in Tris/EDTA buffer was placed in a mare's uterus and left there. The antibiotic in Tris/EDTA solution was left to dissipate naturally by either absorption or being pushed out of the uterus. Some of the control mares were concurrently treated with systemic antibiotics.
No safety concerns were seen in the group treated with ‘PeloMetriSystem,’ whereas one of the control mares had an undesirable reaction to the use of systemic antibiotics. In addition, three mares in the ‘PeloMetriSystem’-treated group became pregnant after treatment, compared to only two in the control group.
Metritis is a problem in humans as well, where it is most commonly referred to as Pelvic Inflammatory Disease (PID). PID is an infection of the uterus (womb), fallopian tubes, and other reproductive organs that causes symptoms such as lower abdominal pain. It is a serious complication of some sexually transmitted diseases (STDs), especially chlamydia and gonorrhea. PID often damages the fallopian tubes and tissues in and near the uterus and ovaries. PID often leads to serious consequences including infertility, ectopic pregnancy (a pregnancy in the fallopian tube or elsewhere outside of the womb), abscess formation, and chronic pelvic pain.
Prompt and appropriate treatment prevents complications of PID caused by scar tissue forming in response to bacterial damage. This scar tissue blocks or interrupts the normal movement of sperm through the fallopian tubes, or eggs into the uterus, and leads to infertility. Treatment with ‘EndoMetriSystem’ will also prove to be useful treatment for PID in humans.
Thus, it is seen that the objects of the present invention are efficiently obtained, although modifications and changes to the invention should be readily apparent to those having ordinary skill in the art, which modifications are intended to be within the spirit and scope of the invention as claimed. It also is understood that the foregoing description is illustrative of the present invention and should not be considered as limiting. Therefore, other embodiments of the present invention are possible without departing from the spirit and scope of the present invention.

Claims

The invention claimed is:

1. A wound healing composition, comprising:

a pharmaceutically acceptable carrier;

an effective amount of an active ingredient of mineral solids, said mineral solids including at least one of a magnesium salt, a potassium salt, a calcium salt, a zinc salt, a rubidium salt, a bromide salt, and a sulfate salt; and

a concentration of methylglyoxal effective to reduce a number of microorganisms at a wound site being added to said active ingredient.

2. The wound healing composition of claim 1, wherein each of the salts includes a pharmaceutically-acceptable counterion.

3. The wound healing composition of claim 1, wherein the active ingredient of mineral solids includes 1.5-75 parts of magnesium ions, 0.5-75 parts of potassium ions, 0.001-10 parts of calcium ions, 0.0001-10 parts of zinc ions, up to 5 parts of rubidium ions, 0.001-20 parts of bromide ions, and up to 20 parts of sulfate ions, said parts expressing a mass of a respective ions divided by a total mass of the mineral solids.

4. The wound healing composition of claim 1, wherein said methylglyoxal added to said active ingredient ranges from 500 to 2000 mg per kg of the composition.

5. The wound healing composition of claim 1, wherein the pharmaceutically-acceptable carrier includes at least one of water, polyethylene glycol, an ointment, and a cream base.

6. The wound healing composition of claim 1, wherein a pH of the composition is between 3.0-7.0.

7. The wound healing composition of claim 1, wherein the wound healing composition is replaced on a wound at least once.

8. A wound healing dressing, comprising:

the wound healing composition according to claim 1; and

a support.

9. The wound healing dressing of claim 8, wherein the support includes at least one of a fibrous gauze material, a hydrogel, a foam, a film, a hydrocolloid, a collagen, and an alginate.