WO2015120310A1

WO2015120310A1 - Method for decolonization of staphylococcus aureus

Info

Publication number: WO2015120310A1
Application number: PCT/US2015/014875
Authority: WO
Inventors: Patrick Schlievert; Aloysius KLINGELHUTZ; Bao Vu
Original assignee: University Of Iowa Research Foundation
Priority date: 2014-02-07
Filing date: 2015-02-06
Publication date: 2015-08-13

Abstract

The present invention relates to methods of preventing the development of diabetes in a patient predisposed to diabetes, or treating complications of diabetes in a diabetic patient, by minimizing the colonization of S. aureus on the skin of the patient. The present invention also provides a method of disrupting a Staphylococcus aureus biofilm on a surface.

Description

METHOD FOR DECOLONIZATION OF STAPHYLOCOCCUS AUREUS

RELATED APPLICATION

This application claims priority to U.S. Provisional Patent Application No. 61/937,426 filed February 7, 2014, the entirety of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

Diabetes mellitus (DM) is a group of metabolic diseases which produce high blood sugar in a patient. There are three major types of Diabetes Mellitus, Type I, Type II, and gestational. Type 1 DM accounts for approximately 5% of all diagnosed cases of diabetes, and results from the body's failure to produce insulin, and presently requires the person to inject insulin or wear an insulin pump. This form was previously referred to as "insulin-dependent diabetes mellitus" (IDDM) or "juvenile diabetes." Risk factors may be autoimmune, genetic, or environmental. There is no known way to prevent type 1 diabetes.

Type 2 DM accounts for about 90% to 95% of all diagnosed cases of diabetes, and results from insulin resistance, a condition in which cells fail to use insulin properly. This form was previously referred to as non insulin-dependent diabetes mellitus (NIDDM) or "adult-onset diabetes." Type 2 diabetes is associated with older age, obesity, family history of diabetes, impaired glucose metabolism, physical inactivity, and race/ethnicity.

Prediabetes is a condition in which individuals have blood glucose or Ale levels higher than normal but not high enough to be classified as diabetes. People with prediabetes have an increased risk of developing type 2 diabetes, heart disease, and stroke. Studies have shown that people with prediabetes who lose weight and increase their physical activity can prevent or delay Type 2 diabetes and in some cases return their blood glucose levels to normal.

Diabetes affects 25.8 million people in the U.S., or about 8.3% of the population. About

18.8 million people are diagnosed with diabetes, and about 7.0 million people are undiagnosed, but have the condition. Diabetes is the leading cause of kidney failure, nontraumatic lower-limb amputations, and new cases of blindness among adults in the United States. It is a major cause of heart disease and stroke, and is the seventh leading cause of death in the U.S.

In 2005-2008, based on fasting glucose or Ale levels, 35% of U.S. adults aged 20 years or older had pre-diabetes (50% of those aged 65 years or older). In 2010 it was estimated 79 million Americans aged 20 years or older had pre-diabetes.

Long-term complications of diabetes are damage to blood vessels, including ischemic heart disease (heart attacks and angina), strokes, peripheral vascular diseases, diabetic retinopathy that can lead to blindness, and diabetic kidney diseases; and neuropathies that lead to numbness, tingling, and pain in the feet. The combination of vascular and neuropathy effects increases the risk of diabetes-related foot problems including diabetic foot ulcers, that may lead to amputations and death.

Accordingly, treatments to prevent complications from diabetes are needed.

SUMMARY OF THE INVENTION

In certain embodiments, the present invention provides a method of preventing the development of diabetes in a patient predisposed to diabetes, comprising contacting skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises: (a) 1% to 10% w/w GML; and (b) 90 to 99% carrier, wherein the carrier comprises (i) propylene glycol, (ii) polyethylene glycol, (iii) hydroxypropyl cellulose, and (iv) an aqueous solution; and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm of contacted skin.

In certain embodiments, the present invention provides a method of treating

complications of diabetes in a diabetic patient, comprising contacting a skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises: (a) 1% to 10% w/w GML; and (b) 90 to 99% carrier, wherein the carrier comprises (i) propylene glycol, (ii) polyethylene glycol, (iii) hydroxypropyl cellulose, and (iv) an aqueous solution; and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm² of contacted skin.In certain embodiments, the present invention provides a method of reducing colonization of S. aureus on a patient's skin where the patient is positively identified as having S. aureus colonized on the patient's skin, comprising contacting the skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises: (a) 1% to 10% w/w GML; and (b) 90 to 99% carrier, wherein the carrier comprises (i) propylene glycol, (ii) polyethylene glycol, (iii) hydroxypropyl cellulose, and (iv) an aqueous solution; and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm² of contacted skin.

In certain embodiments, the present invention provides a method of disrupting a Staphylococcus aureus biofilm on a surface, comprising contacting the surface with an anti-

Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises: (a) 1% to 10% w/w GML; and (b) 90 to 99% carrier, wherein the carrier comprises (i) propylene glycol, (ii) polyethylene glycol, (iii) hydroxypropyl cellulose, and (iv) an aqueous solution; and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm of the surface.

In certain embodiments, the contact time is less than one minute. In certain

embodiments, the contact time is less than 30 seconds. In certain embodiments, the contact time is less than 10 seconds.

In certain embodiments, the step of contacting the skin or surface is repeated at an interval of one to three days. In certain embodiments, the step of contacting the skin or surface is repeated at an interval of two days.

In certain embodiments, the pH of the composition is about 4.5.

In certain embodiments, the PEG has a molecular weight of about 400.

In certain embodiments, the aqueous solution is water or saline.

In certain embodiments, the aqueous solution is at a concentration of 5-25% w/w.

In certain embodiments, the surface is an inanimate surface.

In certain embodiments, the present invention provides an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 comprising (a) 1% to 10% w/w glycerol monolaurate (GML), and (b) 90 to 99% carrier, wherein the carrier comprises (i) about 48.75 to72.75% w/w propylene glycol, (ii) about 25% polyethylene glycol (PEG), (iii) about 1.25% hydroxypropyl cellulose, and (iv) about 1 to 25% aqueous solution.

In certain embodiments, the pH of the composition is about 4.5.

In certain embodiments, the aqueous solution is water or saline.

In certain embodiments, the PEG has a molecular weight of about 400.

In certain embodiments, the present invention provides a cleaning product comprising an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 comprising (a) 1% to 10% w/w glycerol monolaurate (GML), and (b) 90 to 99% carrier, wherein the carrier comprises (i) about 48.75 to72.75% w/w propylene glycol, (ii) about 25% polyethylene glycol (PEG), (iii) about 1.25% hydroxypropyl cellulose, and (iv) about 1 to 25% aqueous solution; and a wiping material.

In certain embodiments, the wiping material is a disposable wipe.

DETAILED DESCRIPTION OF THE INVENTION

Staphylococcus aureus is an anaerobic gram-positive coccal bacterium, also known as "golden staph." Depending on the strain, S. aureus is capable of secreting several exotoxins, which are often associated with specific diseases. S. aureus expresses pyrogenic toxin superantigens (PTSAgs) that induce toxic shock syndrome (TSS). This group includes the toxin TSST-1, enterotoxin type B, which causes TSS associated with tampon use. Other strains of S. aureus can produce an enterotoxin that is the causative agent of S. aureus gastroenteritis. Staph colonizes up to 40% of humans, usually in the anterior nares, but also on other mucosal and skin surfaces. Diabetic patients have increased colonization and infections due to S. aureus.

In certain embodiments, the present invention provides methods of preventing the development of diabetes in a patient predisposed to diabetes, or treating complications of diabetes in a diabetic patient, by minimizing the colonization of S. aureus on the skin of the patient. S. aureus is the most common microbe isolated from diabetic foot ulcers, a common and health-threatening complication of diabetes. Eliminating S. aureus from the skin of the patients prevents injuries from getting infected with S. aureus in diabetic patients.

In certain embodiments, the present invention provides a method of disrupting a

Staphylococcus aureus biofilm on a surface.

In certain embodiments, the present invention is directed to a composition comprising glycerol monolaurate (GML) or a derivative thereof, and a vegetable oil. In one embodiment, the vegetable oil is palm, olive, corn, canola, coconut, soybean, or wheat, or a combination thereof. In a further embodiment, the vegetable oil is present in the composition at about 10% to about 99%, about 20% to about 90%, about 30% to about 80%, or about 40% to about 70%. In one embodiment, the composition comprising GML or a derivative thereof and a vegetable oil further comprises a pharmaceutically acceptable topical carrier, for example, petroleum jelly. In one embodiment, GML or a derivative thereof is present in the composition at a concentration from about 10 μg/mL to about 100 mg/niL, from about 50 μg/mL to about 50 mg/mL, from about 100 μg/mL to about 10 mg/mL, or from about 500 μg/mL to about 5 mg/mL. In another embodiment, the composition comprising GML or a derivative thereof and a vegetable oil further comprises a cellulose derivative, for example either hydroxypropyl cellulose or hydroxyethyl cellulose, or a combination thereof. In a further embodiment, the cellulose derivative is present in the composition up to 1.25% w/w.

In another aspect, the present invention is directed to a composition comprising GML or a derivative thereof, and a gel. In one embodiment, the composition comprising GML or a derivative thereof and a gel has a pH of about 4.0 to about 4.5. In one embodiment, the gel comprises polyethylene glycol, hydroxypropyl cellulose, hydroxyethyl cellulose, or a combination thereof. In a further embodiment, the polyethylene glycol is present at about 25% w/w in the composition. In one embodiment, hydroxypropyl cellulose and hydroxyethyl cellulose are both present in the composition, each at a concentration of about 1.25% w/w. In one embodiment, the GML composition comprising a gel comprises polyethylene glycol with a molecular weight range of about 300 to about 4000. In a further embodiment, the polyethylene glycol has a molecular weight of about 400 or about 1000.

In one embodiment, the GML composition comprising a gel further comprises a topical carrier, e.g., petroleum jelly. In a further embodiment, the composition comprises a vegetable oil.

In one embodiment, the compositions described herein comprise GML or a derivative thereof at a concentration of about 0.001% (w/v) to about 10% (w/v) of the total composition. In a further embodiment, GML or a derivative thereof is present at about 0.005% (w/v) to about 5% (w/v) of the composition. In a further embodiment, GML or a derivative thereof is present at about 0.01 to about 1%. In a still further embodiment, GML or a derivative thereof is present at about 0.1% (w/v) to about 0.5% (w/v) of the composition.

In one embodiment, GML or a derivative thereof is present in the composition at a concentration of about 10 x.§JvaL to about 100 mg/mL. In a further embodiment, GML or a derivative thereof comprises about 50 g/mL to about 50 mg/mL of the composition. In a further embodiment, GML or a derivative thereof comprises about 100 μg mL to about 10 mg/mL. In a still further embodiment, GML or a derivative thereof comprises about 500 μg/mL to about 5 mg/mL.

In one embodiment, the GML composition provided herein comprises propylene glycol at a concentration of about 65% (w/w) to about 80% (w/w). In another embodiment, polyethylene glycol is present in the composition at a concentration of about 20% (w/w) to about 35% (w/w). In one embodiment, both propylene glycol and polyethylene glycol are present in the topical composition.

In one embodiment, the composition comprises a cellulose derivative. In a further embodiment, the composition comprises hydroxypropyl cellulose or hydroxyethyl cellulose. In a yet further embodiment, the cellulose is present at a concentration of about 0.1% (w/w) to about 5.0% (w/w).

In one embodiment, the GML composition comprises an aqueous solvent. In certain embodiments, the aqueous solvent is water, saline, media, or a combination thereof. In certain embodiments, the GML composition is an aqueous wash soluiton.

In one embodiment, the pharmaceutically acceptable topical carrier is petroleum jelly.

In one embodiment, the pH of the GML composition provided herein is from about 4.0 to about 6.0, such as between about 4.0 and 5.5. In some embodiments, the composition provided herein comprises one or more accelerants. In a further embodiment, the accelerant is an organic acid, a chelator, an

antibacterial agent, an anti-fungal agent, an anti-viral agent, or a combination thereof. In a further embodiment, the accelerant is a chelator. In even a further embodiment, the accelerant is EDTA.

In another aspect, the GML composition provided herein has anti-microbial activity. For example, in one embodiment, the composition provided herein is applied topically to humans and other vertebrates, for example for treatment of a bacterial infection such as S. aureus.

Accordingly, in one embodiment, the present invention provides methods for treating a Staphylococcus aureus infection in a subject in need thereof. In one embodiment, the method comprises topically administering to the subject in need thereof, an effective amount of a GML composition provided herein. In one embodiment, the composition comprises GML or a derivative thereof, a vegetable oil, and a pharmaceutically acceptable topical carrier, such as water or saline. In another embodiment, the composition comprises GML or a derivative thereof, a non-aqueous gel, and a pharmaceutically acceptable topical carrier, such as water or saline. In a further embodiment, the composition comprises GML or a derivative thereof, a vegetable oil, a non-aqueous gel, and a pharmaceutically acceptable topical carrier, such as water or saline.

In one embodiment, the compositions disclosed herein are applied topically with the use of a sponge, wipe, or swab.

In one embodiment, the subject has a bacterial infection, where the bacterial infection is a Staphylococcus aureus infection.

As used herein, the term "antimicrobial" means effective in preventing, inhibiting, or arresting the growth or pathogenic effects of a microorganism. "Microorganism" is used herein to mean any bacteria, virus, or fungus. In one embodiment, the formulations of the invention are used to prevent, inhibit, or arrest the growth of Staphylococcus aureus. "Anti-bacterial" as used herein, refers to inhibition or arrest of the growth of a bacterium, a reduction in the severity of or likelihood of developing a bacterial disease, inducing death of the bacterium or reduction or inhibition of the pathogenic effects of the respective bacterium. "Bactericidal" is used interchangeably with "anti-bacterial."

The term "effective amount," as used herein, refers to an amount that is sufficient to effect a beneficial or desired antimicrobial activity, including, without limitation, killing the microorganism or inhibiting microbial infection, growth or toxicity. An effective amount of GML is about 10 μg/mL, about 100 μg mL, about 1 mg/mL, about 10 mg/mL, about 50 mg/mL, or about 100 mg/mL. The terms "treat," "treatment," and "treating" refer to an approach for obtaining beneficial or desired results, for example, clinical results. For the purposes of this invention, beneficial or desired results may include inhibiting or suppressing the growth of a

microorganism or killing a microorganism; inhibiting one or more processes through which a microorganism infects a cell or subject; inhibiting or ameliorating the disease or condition caused by a microbial infection; or a combination thereof. The terms "treat," "treatment," or "treating" also refer to prophylaxis of infection. In some embodiments, the formulations of the invention are used to treat urinary tract infections, vaginal microbial infections, infections of the oral cavities such as those causing gum disease, post-surgical infections including respiratory tract infections, wound or surgical incision site infections, or infections characterized by the production of toxins, including inflammation caused by Staphylococcus aureus infection.

"Prophylaxis," as used herein, can mean complete prevention of an infection or disease, or prevention of the development of symptoms of that infection or disease; a delay in the onset of an infection or disease or its symptoms; or a decrease in the severity of a subsequently developed infection or disease or its symptoms. As used herein, the term "subject" includes humans and other animals. The subject, in one embodiment, is a human.

"Topical," as used herein, refers to the application of the composition to any skin or mucosal surface. "Skin surface" refers to the protective outer covering of the body of a vertebrate, generally comprising a layer of epidermal cells and a layer of dermal cells.

The term "pharmaceutically acceptable topical carrier," as used herein, refers to a material, diluent, or vehicle that can be applied to skin or mucosal surfaces without undue toxicity, irritation, or allergic reaction. A "pharmaceutically acceptable excipient" means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, nontoxic and neither biologically nor otherwise undesirable, and includes an excipient that is acceptable for veterinary use as well as human pharmaceutical use. A "pharmaceutically acceptable excipient" as used in the present application includes both one and more than one such excipient.

The term "accelerant," as used herein, refers to a compound, substance, liquid, powder, or mixture that, when added to the composition, has the effect of enhancing or contributing to the antimicrobial properties of the composition. Accelerants may be an organic acid including, without limitation, lactic acid, ascorbic acid, citric acid, formic acid, benzoic acid, and oxalic acid. The accelerant, in another embodiment, is a chelator, and in one embodiment, is selected from ethylenediaminetetraacetic acid (EDTA), dimercaprol, dimercaptosuccinic acid (DMSA), 2,3-dimercapto-l-propanesulfonic acid (DMPS), alpha lipoic acid (ALA), or combinations thereof. In another embodiment, the accelerant is an antibiotic agent. Antibiotics for use with the invention, for example, include aminoglycosides, carbacephems, cephalosporins, glycopeptides, lincosamides, lipopetides, macrolides, monobactams, nitrofurans, penicillins, polypetides, quinolones, sulfuramides, and tetracyclines.

The term "biofilm," as used herein, means an aggregate of microorganisms, usually bacterial, adhered to one another and growing on a surface. The microbial cells in the biofilm typically produce an extracellular matrix known as an extracellular polymeric substance. Often, this matrix and the density of the aggregate itself significantly increase the antibiotic resistance of the bacteria in the biofilm. Biofilms can be involved in UTIs, ear infections, and dental diseases such as gingivitis, and can also form on the surface of implanted devices including prostheses, catheters, or heart valves.

In one embodiment, the composition provided herein comprises the monoglyceride GML. GML is a fatty acid ester of glycerol, derivative of lauric acid, with the chemical formula C15H30O4. GML is also known in the art as glyceryl laurate or monolaurin. GML is found naturally in breast milk and some plants, and is used as a food and cosmetic additive. GML and other glycerides are listed in the Generally Recognized as Safe Substances database by the US Food and Drug Administration. GML and related compounds have been previously disclosed in US patent application Nos. 10/579,108 (filed November 10, 2004) and 11/195,239 (filed August 2, 2005), the disclosures of each of which are herein incorporated by reference for all purposes.

GML can be synthesized in multiple forms including both R and S optical isomers, as well as forms with lauric acid in the 1/3-position and in the 2-position. The composition provided herein, in one embodiment, comprises the R isomer of GML. In another embodiment, the composition provided herein comprises the S isomer of GML. In yet another embodiment, a racemic mixture of isomers is provided in the composition.

The compositions of the present invention can be prepared as described in US Patent

Publication 2013/0281532, which is incorporated by reference in its entirety herein.

In one embodiment, the method comprises administering to the subject a topical composition comprising GML or a derivative thereof, as described herein. In one embodiment, the method comprises topically administering to the subject an effective amount of a

composition comprising GML or a derivative thereof, a vegetable oil, and a pharmaceutically acceptable topical carrier. In another embodiment, the method comprises topically administering an effective amount of a composition comprising GML, a non-aqueous gel, and a

pharmaceutically acceptable topical carrier. In one embodiment, the method of treating a microbial infection comprises applying an effective amount of one or more of the GML compositions described herein to at least one skin or mucosal surface of a subject.

In some embodiments, the composition is applied to or impregnated in a wipe, sponge, swab, or other material, and then applied to the skin or mucosal surface of the subject using the respective material. As used herein, the term "swab" refers to a material suitable for applying a liquid, gel, wax, cream, or lotion to a skin or mucosal surface, or the act of applying a liquid, gel, wax, cream, or lotion to the skin or mucosal surface, or the act of collecting a liquid, gel, wax, cream, lotion, or fluid from the skin or mucosal surface. In some embodiments, the material is attached to a holder, for example a stick, wire, rod, or applicator. In further embodiments, the material attached to a holder is attached at one or both ends thereof. In some embodiments, the wipe, sponge, swab, or other material is pre-loaded or packaged together with the composition. EXAMPLE

There are studies that suggest that persons with either Type I or Type II diabetes are at increased risk of bacterial infection due to the vascular and nerve damage and complicated by elevated blood sugar. One of the most often seen and serious infections is that due to

Staphylococcus aureus. This organism colonizes up to 40% of human, usually in the anterior nares, but also on other mucosal and skin surfaces (1 1, 13, 15). It has also been shown that diabetic patients have increased colonization and infections due to S. aureus (1, 3, 4, 6-10, 16, 18, 19).

We evaluated two persons, both women, with Type I and two persons with Type II diabetes for the presence of S. aureus. The two persons with Type I diabetes appeared healthy, but based on swabbing their palm and forearm skin with swabs pre-wetted with 0.1 ml of normal saline these persons had approximately 10 S. aureus on their total skin surfaces (based on estimates of total skin area); the anterior nares of these persons were not evaluated for S. aureus. One of the persons was colonized with a community-associated methicillin-resistant S. aureus (CA-MRSA), whereas the other person was colonized with a methicillin-sensitive S. aureus (MSSA). The CA-MRSA was a USA400 clonal group S. aureus based on its production of the superantigen staphylococcal enterotoxin C (2), whereas the MSSA was a USA200 strain based on its production of the superantigen toxic shock syndrome toxin- 1 (17). The two persons, both males, with type II diabetes were colonized on the skin and anterior nares with S. aureus, both of which were shown to be CA-MRSA, probably of the USA400 clonal group based on production of the superantigen enterotoxin C. Both of these persons ultimately succumbed, one from an overwhelming infection due to his colonizing strain of S. aureus and one from an unknown cause. The numbers of S. aureus on the skin of these persons was estimated to be approximately 10¹⁰ and in the anterior nares to be approximately 10⁶.

Previous research has shown that glycerol monolaurate (GML) alone kills all S. aureus strains (54 tested) at GML concentrations of <500 μg/ml (5, 12, 14). Additionally, at sub- growth-inhibitory concentrations, GML prevents production of exotoxins (5, 12, 14). We have prepared GML in a Skin/Mucosal Gel (S/M Gel) that potentially can be used to decolonize diabetic persons of S. aureus, either as S/M Gels (or GML washes). The S/M Gels have the composition listed in Table 1. The action of this gel results in synergy among GML, the nonaqueous gel, and the resultant pH of 4.5.

Table 1. Composition of S/M Gel containing GML

Ingredient Supplier Function %w/w

Propylene glycol USP Gallipot Lubricant 69.75%

Polyethylene glycol 400 NF Gallipot Lubricant 24%

Hydroxypropyl cellulose Gallipot Gelling agent 1.25%

Distilled water Baxter Healthcare Diluent 5.0%

Total 100%

Glycerol Monolaurate Colonial Active ingredient 0.5%, 5%, 10% *pH 4.5 via pH paper

We tested the ability of these gels to kill 5. aureus MSSA strain MN8 (USA200), CA- MRSA strain MW2 (USA400), and CA-MRSA LAC (USA300). All organisms were cultured overnight in Todd Hewitt broth (Difco Laboratories, Detroit, MI) and then adjusted to approximately 2 x 10⁹ colony-forming units (CFUs)/ml in Todd Hewitt broth. From this diluted stock, 0.1 ml was added to 0.9 ml of each of the three GML S/M Gels (0.5%, 5.0%, and 10%); this gave bacterial concentrations of approximately 2 x 10⁸/ml of GML S/M Gels. The cultures in triplicate were incubated stationary in a 5% C0₂ incubator. At intervals of 30 seconds, 1 minute, 15 minutes, 1 hour, 4 hours, and 24 hours, 0.1 ml samples of all cultures were spread onto sterile sheep blood agar plates (BAPs) for CFUs/ml determination. Control cultures in triplicate consisted of the same dilution of S. aureus strains in 1 ml of TH broth. The data are presented in Table 2. Table 2. Inhibition of S. aureus strains by GML S/M Gels compared to Todd Hewitt broths.

Note: Lower limit of detection was 10 CFUs/ml

Whereas typical growth of S. aureus occurred in all THB cultures over the 24 hour test period, as early as 30 seconds after inoculation, no viable S. aureus strains were cultured from the GML (0.5%, 5%, and 10%) S/M Gels. These data indicate that GML Gels are strongly antimicrobial for all three S. aureus strains on contact, requiring less than 30 seconds for complete killing. Additionally, there was no out-growth of S. aureus from strains incubated with gels for 24 hours.

As an additional control, 0.1 ml of GML (5%) S/M Gel was plated onto each of 3 BAPs and incubated for 5 minutes, noting that GML Gels dilute rapidly in media. Then, 0.1 ml of each S. aureus strain was plated onto those three BAPs. After 24 hours incubation, all three plates contained lawns of S. aureus, indicating the inhibition of growth seen above was not the result of carry-over of GML S/M Gels onto the BAPs; dilution of the GML (5%) Gel with the approximately 30 ml of media in the BAPs (1/300 dilution; GML concentration 167 μg/ml) removed its antimicrobial activity for these strains.

One additional experiment was performed. Three persons were identified who were positive for S. aureus on their hands (Table 3) as tested by using swabs pre-wetted with 0.1 ml of normal saline, rolling these swabs on the skin surfaces, and plating dilutions on BAPs. The three persons had approximately 15,000 CFUs of S. aureus on their hands. The persons then washed their hands with GML (5%) S/M Gel for 1 minute. Subsequently, each person was reevaluated for & aureus. There was a highly significant reduction in S. aureus CFUs by washing hands with GML (5%) S/M Gel (p=0.019 by paired Student's t test). The reduction was such that no CFUs were detected on any of the three subjects.

Table 3. Effect of GML (5%) S/M washing for 1 minute on hand colonization with S. aureus.

Collectively, these data indicate that GML S/M Gels may be highly effective washes to reduce S. aureus on diabetic and healthy human skin and mucous membranes.

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18. Tuazon, C. U., A. Perez, T. Kishaba, and J. N. Sheagren. 1975. Staphylococcus aureus among insulin-injecting diabetic patients. An increased carrier rate. JAMA 231 : 1272. 19. Ulbrecht, J. S., P. R. Cavanagh, and G. M. Caputo. 2004. Foot problems in diabetes: an overview. Clinical Infectious Diseases 39 Suppl 2:S73-82.

Although the foregoing specification and examples fully disclose and enable the present invention, they are not intended to limit the scope of the invention, which is defined by the claims appended hereto.

All publications, patents and patent applications are incorporated herein by reference. While in the foregoing specification this invention has been described in relation to certain embodiments thereof, and many details have been set forth for purposes of illustration, it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments and that certain of the details described herein may be varied considerably without departing from the basic principles of the invention.

The use of the terms "a" and "an" and "the" and similar referents in the context of describing the invention are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms "comprising,"

"having," "including," and "containing" are to be construed as open-ended terms (i.e., meaning "including, but not limited to") unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

Embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein.

Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

WHAT IS CLAIMED IS:

A method of preventing the development of diabetes in a patient predisposed to diabetes, comprising contacting skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises:

(a) 1 % to 10% w/w GML; and

(b) 90 to 99% carrier, wherein the carrier comprises

(i) propylene glycol,

(ii) polyethylene glycol,

(iii) hydroxypropyl cellulose, and

(iv) an aqueous solution;

3 2 and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm of contacted skin.

A method of treating complications of diabetes in a diabetic patient, comprising contacting a skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises:

(a) 1 % to 10% w/w GML; and

(b) 90 to 99% carrier, wherein the carrier comprises

(i) propylene glycol,

(ii) polyethylene glycol,

(iii) hydroxypropyl cellulose, and

(iv) an aqueous solution;

and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm of contacted skin.

A method of reducing colonization of S. aureus on a patient's skin where the patient is positively identified as having S. aureus colonized on the patient's skin, comprising contacting the skin of the patient with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises:

(a) 1 % to 10% w/w GML; and

(b) 90 to 99% carrier, wherein the carrier comprises

(i) propylene glycol,

(ii) polyethylene glycol,

(iii) hydroxypropyl cellulose, and

(iv) an aqueous solution;

and wherein colonization of S. aureus is reduced to less than 10 3 CFU/2.5 cm 2 of contacted skin.

4. A method of disrupting a Staphylococcus aureus biofilm on a surface, comprising contacting the surface with an anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 for less than two minutes, wherein the composition comprises:

(a) 1 % to 10% w/w GML; and

(b) 90 to 99% carrier, wherein the carrier comprises

(i) propylene glycol,

(ii) polyethylene glycol,

(iii) hydroxypropyl cellulose, and

(iv) an aqueous solution;

3 2 and wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm of the surface.

5. The method of any one of claims 1-4, wherein the contact time is less than one minute.

6. The method of any one of claims 1-4, wherein the contact time is less than 30 seconds.

7. The method of any one of claims 1-4, wherein the contact time is less than 10 seconds.

8. The method of any one of claims 1-7, wherein the step of contacting the skin or surface is repeated at an interval of one to three days.

9. The method of any one of claims 1-7, wherein the step of contacting the skin or surface is repeated at an interval of two days.

10. The method of any one of claims 1-9, wherein the pH of the composition is about 4.5.

1 1. The method of any one of claims 1-9, wherein the PEG has a molecular weight of about 400.

12. The method of any one of claims 1-9, wherein the aqueous solution is water or saline.

13. The method of any one of claims 1-9, wherein the aqueous solution is at a

concentration of 5-25% w/w.

14. The method of any one of claims 4-13, wherein the surface is an inanimate surface.

15. The method of any one of claims 1-14, wherein colonization of S. aureus is reduced to less than 10 CFU/2.5 cm²

16. An anti-Staphylococcus aureus composition having a pH of 4.0 to 6.0 comprising

(a) 1% to 10% w/w glycerol monolaurate (GML), and

(b) 90 to 99% carrier, wherein the carrier comprises

(i) about 48.75 to72.75% w/w propylene glycol,

(ii) about 25% polyethylene glycol (PEG),

(iii) about 1.25% hydroxypropyl cellulose, and

(iv) about 1 to 25% aqueous solution.

17. The anti-Staphylococcus aureus composition of claim 16, wherein the pH of the composition is about 4.5.

18. The anti-Staphylococcus aureus composition of any one of claims 16-17, wherein the aqueous solution is water or saline.

1 . The anti-Staphylococcus aureus composition of any one of claims 16-18, wherein the aqueous solution is at a concentration of 5-25% w/w.

20. The anti-Staphylococcus aureus composition of any one of claims 16-1 , wherein the PEG has a molecular weight of about 400.

21. A cleaning product comprising the anti-Staphylococcus aureus composition of claim 16 and a wiping material.

22. The cleaning product of claim 21, wherein the wiping material is a disposable wipe.