EP2389069A1

EP2389069A1 - Methods and compositions for the treatment and prevention of infections

Info

Publication number: EP2389069A1
Application number: EP10733709A
Authority: EP
Inventors: Jeffrey P. Gilbard; Robert B. Huson
Original assignee: Advanced Vision Research LLC
Current assignee: Advanced Vision Research LLC
Priority date: 2009-01-20
Filing date: 2010-01-19
Publication date: 2011-11-30
Also published as: KR20110114674A; AU2010207010A1; CA2750084A1; WO2010085323A1

Abstract

The instant invention provides preparations comprising activated chlorine dioxide that improves the antibacterial effect of the oxidizing antimicrobial agent preparation. The invention has particular use as an eye care preparation such as an eye drop. The invention further provides methods for reducing bacterial colonization and treating infection.

Description

METHODS AND COMPOSITIONS FOR THE TREATMENT AND PREVENTION OF INFECTIONS

RELATED APPLICATION This application claims the benefit of and priority to U.S. Provisional Patent

Application No. 61/205,254, filed January 20, 2009. The contents of the application is incorporated herein by reference.

BACKGROUND OF THE INVENTION A condition known as dry eye causes chronic eye irritation resulting from decreased tear production or increased evaporation that results in a loss of water from the tear film and an increase in tear film osmolarity. This increase in tear film osmolarity results in an osmotic dehydration of the surface associated with a decrease in the density of conjunctival goblet cells. Recently it has been shown that dry eye patients have increased bacterial colonization of their eyelids, and that the bacteria found in these patients decrease the proliferation of conjunctival goblet cells in tissue culture (Graham et al Analysis of Bacterial Flora in Dry Eye, Ocular Surface, 3(1):S68, 2005). Dry-eye patients are also known to be more prone to eye infections, especially in the context of contact lens wear (Lemp MA "Is the dry eye contact lens wearer at risk?", Cornea (United States), 1990, 9 Suppl 1 pS48-50; discussion S54).

Punctal plugs are a frequently used treatment for dry eye. They provide symptomatic relief for patients with dry eye, reduce elevated tear film osmolarity in the disease and reduce ocular surface staining. A problem with punctal plugs is that they are frequently colonized by pathogenic noncomensals, including Pseudomonas aeruginosa and Staphylococcus aureus, that may cause symptoms and increase the risk of eye infections (Soukiasian SH Microbiology of Explanted Punctal Plugs, ARVO Annual Meeting, Program#/Poster# 4981/B305, April 29, 2004; Sugita J, Yokoi N, Fullwood NJ, et al. "The detection of bacteria and bacterial biofilms in punctal plug holes", Cornea (United States), May 2001 , 20(4) p362-5). Another condition of clinical significance is inflammation of the eyelids and the eye surface. This inflammatory disorder, frequently resulting in symptoms of eye irritation, is called blepharitis. In a study involving 332 patients with blepharitis and 160 normal controls, it has been shown that blepharitis patients have greater quantities of bacteria on their eyelids compared to normal controls. This finding applied to patients with both anterior and posterior blepharitis (Groden LR, Murphy B, Rodnite J, et al. "Lid flora in blepharitis", Cornea (United States), Jan 1991, 10(1) p50-3). Bacterial overgrowth has been hypothesized to contribute to the symptoms of blepharitis by the production of bacterial lipases and esterases that hydrolyze the wax and sterol esters in meibum, creating free fatty acids that are irritating to ocular tissue and may effect tear film stability (Ta CN, Shine WE, McCulley JP, et al., Cornea (United States), Aug 2003, 22(6) p545-8). In addition, these fatty acids may promote eyelid and ocular surface inflammation (Shine WE, McCulley JP, Pandya AG Exp Eye Res (England), Apr 2003, 76(4) p417-20).

Eye hygiene, including eye lid cleaning, has been recommended for all of these conditions or circumstances by eye doctors. Many of the treatments available for dry eye conditions are formulated as drops or ointments. However, drops and ointments have a limited lifespan once opened as they commonly become infected with microbial growth.

Chlorine dioxide has been called the ideal biocide. Despite the many advantages of chlorine dioxide, commercial use is limited because it is an unstable, highly reactive gas which is soluble in and decomposes in water. See, e.g., U.S. Pat. No. 4,941,917. Therefore, it has heretofore been necessary to generate aqueous chlorine dioxide solutions on site for immediate use or use within a relatively short time (typically less than an hour). Due to these limitations, chlorine dioxide has not met its full potential as an antimicrobial agent. Similarly, other oxidizing antimicrobial agents such as sodium perborate have been under utilized because of stability and effectiveness issues. Therefore, a need exists for improved preparations that can be used, for example, in or around the eye and that are stable for extended periods of time.

SUMMARY OF THE INVENTION

The instant invention is based on the discovery that eye care preparations comprising an activated oxidizing antimicrobial agent such as chlorine dioxide exhibit beneficial antimicrobial effects. For example, preparations comprising activated chlorine dioxide are useful for the treatment and prevention of microbial infection and for the preservation of ocular products. The addition of chlorine dioxide preserves the solution for extended periods of time. Moreover, these preparations resist microbial growth prior to and after being used by an individual.

Accordingly, in at least one aspect, the instant invention provides ophthalmologically suitable compositions comprising activated chlorine dioxide, wherein the osmolality of the composition is less than 200 mOsmol/Kg.

A specific ophthalmologically suitable composition of the invention comprises activated chlorine dioxide, wherein the osmolality of the composition is less than 180 mOsm/Kg. The cholorine dioxide can be present in a concentration of about 10-125 ppm, of about 25-75 ppm or about 50 ppm. The ophthalmologically suitable composition can further comprise a weak acid, such as boric acid. The ophthalmologically suitable composition can further comprise a buffer component, e.g., an inorganic buffer component such as borate, phosphate, or carbonate, or combinations thereof.

The ophthalmologically suitable compositions of the invention have a final pH of about 6 to about 10, from 6.5 to about 8, or of about 7.2.

The ophthalmologically suitable compositions, can also contain a pharmaceutically acceptable carrier.

Some of the ophthalmologically suitable compositions of the invention contain trehalose. The trehalose can be present in a range of 2.5-5%. Exemplary final concentrations of trehalose can be 2.5%, 3.0%, 3.5%, 4.0%, 4.5% or 5.0%. One specific ophthalmologically suitable composition has a final concentration of 3.4% trehalose.

The ophthalmologically suitable compositions of the invention can be solutions, creams, pastes, ointments, or gels. The ophthalmologically suitable compositions of the invention can also contain an opthalmoligically acceptable thickening agent, such as carboxymethylcellulose, dextran, sorbitol, mannitol, ethyl cellulose, hydroxytheyl cellulose, hydroxymethyl cellulose, hydroxypropyl methylcellulose, polyvinyl pyrrolidine, polyvinyl alcohol, carboxyvinyl polymer, carbopol, isopropyl myristate, petroleum jelly, mineral oil, lanolin, polyethylene glycol, propylene glycol, glycerin, glucose, sucrose, lactose, agar, alcohols, gums, waxes, or combinations thereof. A specific ophthalmologically suitable composition of the invention contains an opthalmoligically acceptable thickening agent at a final concentration of about 2.5%, 2%, 2.25%, 2.5%, 1.75%, 1.5%, 1.25%, 1.0%, 0.75%, 0.5%, and 0.25%. In a preferred embodiment, the composition comprises the opthalmoligically acceptable thickening agent sodium carboxymethylcellulose at a final concentration of 0.25%.

The ophthalmologically suitable compositions of the invention are suitable for application to the eye, eye-lid, eye margin, punctal plug or a contact lens.

The ophthalmologically suitable compositions of the invention do not contain a bicyclic compound, e.g., dyphilline.

The ophthalmologically suitable compositions of the invention can also contain boric acid. One exemplary ophthalmologically suitable composition of the invention has an osmolality of 180 mOsm/Kg or less and wherein the chlorine dioxide is present in a concentration of about 50-75 ppm.

One specific ophthalmologically suitable composition of the invention contains the following components at about the following final concentrations:

Sodium Carboxymethylcellulose 0.2500%

Sodium Chloride 0.03777%

Potassium Chloride 0.01244%

Magnesium Chloride, Hexahydrate, 0.00085%

Sodium Phosphate, Monobasic, „ _nnnQ,_0/ Monohydrate

Calcium Chloride, Dihydrate 0.001 10%

Sodium Bicarbonate 0.01868%

Boric Acid, NF 0.15000%

Sodium Borate, Decahydrate 0.00008%

Chlorine Dioxide 0.00500%, and

Trehalose 3.420%.

This specific ophthalmologically suitable composition has an osmolality of about 145-155 mOsm/Kg.

In a specific embodiment, ophthalmologically suitable compositions of the invention are aqueous compositions. The invention also provides methods for treating microbial colonization or infection in or around the eye of a subject by applying the ophthalmologically suitable compositions of the invention to the colonized or infected area in an amount sufficient to treat a microbial colonization or infection in a subject. Exemplary microbial colonizations include bacterial, fungal or yeast colonization or infection. The ophthalmologically suitable compositions can be in the form of drops, solutions, creams, pastes, ointments, or gels. They can also be formulated into sustained-release carriers, such as sustained release polymers, liposomes or microcapsules. The invention also provides methods of preventing or treating an infection of the ocular surface in a subject by applying a composition of the invention to an ocular surface in an amount sufficient to prevent or treat an infection of the ocular surface in the subject. An exemplary infection of the ocular surface includes bacterial conjunctivitis. The invention also provides a method for reducing the risk of infection in patients, including dry-eye patients, wearing contact lenses.

The invention also provides methods of reducing the risk of infection of the eye in an eye surgery patient by applying the compositions described herein to an eyelid or ocular surface prior to a surgical procedure in an amount sufficient to reduce the risk of infection in the eye of a surgical patient. In accordance with this method, the composition is applied multiple times over a number of days preceding the surgery.

The invention also provides methods of reducing the risk of infection of the eye in a subject wearing a punctal plug by applying the compositions described herein to the eye surface, the eyelid margin, the eyelid or the punctal plugs or into the tear film in an amount sufficient to reduce the risk of infection in the eye of the subject wearing a punctal plug.

The invention also provides methods for treating or reducing the risk of infection in a subject by applying the topical composition described herein to the area that is infected or at risk of becoming infected in an amount sufficient to treat or reduce the risk of infection.

The invention also provides methods of reducing colonization or of treating an infection of the mucosal tissue or the epithelial tissue in a subject by applying the composition described herein to the mucosal tissue or the epithelial tissue in an amount sufficient to treat the infection in the subject. The invention also provides a method of treating dry eye in a subject by applying the eye care composition described herein to the eye or eyelid in an amount sufficient to treat dry eye in the subject. The invention also provides a method of increasing goblet cell density and mucosal epithelial membrane mucin expression in a subject by applying the eye care composition described herein to the eye in an amount sufficient to increase goblet cell density and mucosal epithelial membrane mucin expression. This increase can be determined using rose Bengal or other vital staining techniques. In dry eye, the ocular surface stains less with an increase in conjunctival goblet cell density and mucosal epithelial membrane mucin expression.

The invention also provides methods for sterilizing a surface by contacting the surface with the composition described herein, or incorporating within a surface the composition described herein, in an amount sufficient to sterilize the surface.

Exemplary surfaces include surfaces on a body, a medical instrument, or a medical device. An exemplary medical device includes a contact lens.

The invention also provides kits for the treatment or prevention of a microbial infection, comprising the compositions described herein and instruction for use. The kits may further comprise an applicator.

DETAILED DESCRIPTION OF THE INVENTION

At present, there exists a need for compositions and methods for preventing microbial growth and treating microbial infections. In particular, there exists a need for compositions for preventing microbial growth in products for use in the eye or surrounding area. In particular, the need exists for compositions that remain free of microbial growth for extended periods of time, i.e., compositions that contain a preservative that inhibit microbial growth and that do not irritate the area to which it is applied. Preferably, the compositions will contain an antimicrobial composition that will kill or retard the growth of microbes (e.g., a bactericidal or bacteriostatic composition). In certain embodiments, the compositions are also useful for treating or preventing infection of, for example, the skin. In further embodiments, the preparations of the invention are useful in treating or preventing infection on surfaces, medical devices, and medical instruments.

Definitions

The invention will be described with reference to the following definitions that, for convenience, are collected here. The term "cleaning an eyelid" is used herein to describe the act of significantly reducing the amount of dirt, debris, or bacteria, from an eyelid.

The term "dry eye" is known in the art as a condition of a subject that has a loss of water from the tear film due to either a decrease in tear production or an increase in tear film evaporation. Tear production can decrease from lacrimal gland disease, including, but not limited to, that which occurs in Sjogren's syndrome, or from anything that decreases corneal sensation. Examples of conditions that decrease corneal sensation include, but are not limited to, diabetes, long-term contact lens wear, and corneal surgery, including LASIK eye surgery. Tear film evaporation can increase from meibomian gland dysfunction, that manifests itself by stenosis or closure of the meibomian gland orifices, or in the presence of large palpebral fissure widths. Causes for large palpebral fissure width include, but are not limited to, normal biological variation and thyroid eye disease. Dry eye is often an age related disease, and may also be caused by a dietary deficiency of omega- 3 essential fatty acids. Dry eye is associated with bacterial overcolonization of the eyelids.

The term "eyelid" as used herein, includes the tarsal conjunctival surface, both the interior and exterior surfaces of the eyelid, the eyelid margin, the glands in and around the eyelid margins, the hair follicles of the eyelid, the eyelashes, and the periocular skin surrounding the eye. The term "eye surface inflammatory disorder," as used herein, is intended to include disorders associated with eye surface inflammation. These disorders include dry eye, where ocular surface inflammation has been demonstrated, as well as both anterior and posterior blepharitis. In anterior blepharitis, the inflammation is centered around the eyelashes. Posterior blepharitis or meibomitis is associated with inflammation of the tarsal and bulbar conjunctiva, and complicated by hordeolums and chalazions, and leads to meibomian gland dysfunction. Both anterior and posterior blepharitis are associated with bacterial overcolonization of the eyelids.

Animal models with combined dry eye and eye surface inflammatory disorder have been produced, and can be used to test the efficacy of the antimicrobial preparations provided herein. For example, a rabbit model for meibomianitis and meibomian gland dysfunction has been developed. In this animal model, meibomian gland orifice closure results in the development of inflammation around the meibomian glands (i.e., meibomianitis), inflammation in the eyelids (blepharitis), inflammation in the conjunctiva (conjunctivitis) and in an increase in tear film osmolality and a decrease in the levels of corneal glycogen and conjunctival mucus- containing goblet cells characteristic of dry-eye surface disease.

The term "eyelid disorder" is defined as a disorder that results in inflammation of the eyelashes and/or eyelash follicles and/or eyelid margins, or inflammation of the lipid producing glands that are located in the eyelid, including meibomianitis and anterior blepharitis. Exemplary eyelid disorders include, but are not limited those caused by bacterial infection.

The term "ocular disorder" as used herein, includes ocular surface disorders, disorders of the eyeball, periocular skin disorders, and eyelid disorders. Exemplary ocular disorders include, but are not limited to, dysfunctions of the tear film, inflammation of the eyelid margins due to bacterial infection, infections inside the eye known as endophthalmitis, and dry eye.

The term "treatment" as used herein is defined as prophylactic treatment (e.g., daily preventative use) or therapeutic treatment (e.g., a single treatment or a course of treatment) of a subject with or at risk for an ocular disorder, or with an ear or skin condition, that are associated with or exacerbated by infections or bacterial colonization.

The term "preparation or antimicrobial preparation" as used herein includes compositions comprising an oxidizing antimicrobial compound, for example, chlorine dioxide. The preparation of the invention can be a solution, cream, paste, ointment, gel or the like. The preparations of the invention can be applied to, for example, the skin, eye, or eyelid.

As used herein the language "pharmaceutically acceptable carrier" is intended to include any and all solvents, agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the antimicrobial preparations described herein, such media can be used in the compositions of the invention. Pharmaceutical compositions suitable for topical application preferably take the form of a drop, solution, ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Exemplary carriers which may be used include water, carboxymethylcellulose, petroleum jelly, mineral oil, lanolin, polyethylene glycols, alcohols, and combinations of two or more thereof. As used herein, the term "ophthalmologically suitable composition" is intended to mean that the composition is appropriate for application to the eye and the area around the eye. In certain embodiments, this means that the composition does not irritate or inflame the eye. Moreover, it is intended to mean that application to the eyelid or surrounding areas does not result in discomfort to the subject.

As used herein, the term "activated chlorine dioxide" is intended to mean chlorine dioxide that has been produced in the presence of a strong acid thereby increasing the ability of the chlorine dioxide to be bactericidal or bacteriostatic. In specific embodiments, the chlorine dioxide composition is contacted produced in a solution that has a pH of less than 6.8, 6.7, 6.6, 6.5, 6.4, 6.2, 6.1, 6.0, 5.9, 5.8, 5.5, 5.3, 5.1, 4.9, 4.7, 4.5 or lower. Although the pH of the composition may ultimately be increased, the chlorine dioxide is activated once upon the pH of the solution being adjusted to a sufficiently acidic pH.

Methods and Compositions

Effective health and cleanliness of an eye is dependant upon the ability to control the level of microbes. Accordingly, compositions that have extended utility once opened, i.e., that resist microbial growth, are useful for the treatment of the eye. The present invention provides compositions and methods, which decrease, e.g., significantly decrease, the number of microbes present in or around, for example, an eye, or in materials used in or around the eye.

Accordingly, the invention is directed to a preparation comprising an antibacterial concentration of activated chlorine dioxide, wherein the composition has an osmolality of less than 200 mθsmol/kg. The preparation may also contain a pharmaceutically acceptable carrier or water. The preparation may be used as a preservative for materials used in conjunction with the eye such as eye drops or may be specifically formulated for the treatment of a particular disorder, e.g., an ocular disorder selected from blepharitis, dry eye, infectious conjunctivitis, an ear infection, or a skin infection. The preparation may also be used to sterilize, for example, surgical instruments, medical indwelling devices, surfaces and the like. The preparation may also be incorporated into the surface of medical devices for sustained release of the preparation. One of skill in the art would understand that the preparation of the invention may be prepared in the form of drops, solution, paste, cream, foam, gel, ointment, or the like, or incorporated into sustained-release carriers such as sustained-release polymers, liposomes and microcapsules.

There are several commercial generators for producing the chlorite/chlorine chlorine dioxide. Suitable generators are disclosed in U.S. Pat. Nos. 4,247,531 ; 5,204,081 ; 6,468,479; and 6,645,457, the disclosures of which are incorporated herein by reference.

Chlorine dioxide can be produced with high efficiency by reducing sodium chlorate in a strong acid solution with a suitable reducing agent (for example, hydrogen peroxide, sulfur dioxide, or hydrochloric acid): 2ClO₃ ^" + 2Cl^" + 4H⁺ → 2ClO₂ + Cl₂ + 2H₂O

Alternatively, chlorine dioxide can be made by one of three methods using sodium chlorite: The sodium chlorite-chlorine gas method (2 NaClO₂ + Cl₂ -→ 2 ClO₂ + 2 NaCl); the sodium chlorite-hypochlorite method (2 NaClO₂ + 2 HCl + NaOCl → 2 ClO₂ + 3 NaCl + H₂O); or the sodium chlorite-hydrochloric acid method (5 NaClO₂ + 4 HCl → 5 NaCl + 4 ClO₂ ). Finally, chlorine dioxide can be produced by electrolysis of a chlorite solution (NaClO₂ + H₂O → ClO₂ + NaOH + 1/2 H₂). Preparations of the invention comprise between about 25-200 ppm of chlorine dioxide, about 50-150 ppm of chlorine dioxide, about 50-100 ppm of chlorine dioxide, or about 50-75 ppm of chlorine dioxide. For treatment of infection, higher values may be used.

In certain embodiments, these chemical reactions take place in an acidic environment in order to activate the chlorine dioxide.

In certain embodiments, the antimicrobial preparation is an aqueous solution containing chlorine dioxide as described herein. The solutions of the invention can have an osmolality of, for example, about 180 mOsm/Kg, about 175 mOsm/Kg or less, about 170 mOsm/Kg, about 165 mOsm/Kg or less, about 160 mOsm/Kg or less, or about 155 mOsm/Kg or less.

An exemplary preparation of the invention is a preparation having an osmolality of 165 mOsm/Kg or less and a chlorine dioxide concentration of about 50- 75 ppm.

The preparations may further include buffers, solubilizers, viscosity increasing agents, preservatives, anti-inflammatory agents and salts. In a preferred embodiment, the invention provides an ophthalmologically suitable composition comprising activated chlorine dioxide, wherein the osmolality of the composition is less than 180 mOsm/Kg. The eye drop includes a balance of electrolytes found in natural tear fluid required for ocular surface maintenance, function and repair. These electrolytes are present in amounts and proportions sufficient to maintain or restore conjunctival goblet cells and corneal glycogen, thereby maintaining mucus-mediated lubrication and the potential for normal healing. This enables topical application of the composition to ocular surfaces preferably without substantially reducing the density of conjunctival mucus-containing goblet cells or levels of corneal glycogen. Goblet cells form a critical layer of the tear film, providing the eye surface with lubrication, and playing an important role in the system that traps foreign matter that may enter the eye, and promptly removes it. Corneal glycogen is the energy source for the sliding step in corneal wound healing. Their preservation is therefore important in maintaining the health of ocular surfaces. The ophthalmologically suitable compositions of the invention include, in addition to chlorine dioxide, a balance of electrolytes naturally found in tear fluid. These electrolytes principally include major amounts of sodium and chloride, and lesser amounts of potassium and bicarbonate.

The ophthalmologically suitable compositions may also contain other naturally-occurring elements of the tear fluid, such as proteins, enzymes, lipids and metabolites as described in U.S. Patent No. 4,91 1,933. Typically, in an isotonic preparation, the potassium is present at a concentration of about 22.0 to 43.0 mM/1, the bicarbonate is present at a concentration of about 29.0 to 50.0 mM/1, the sodium is present at a concentration of about 130.0 to 140.0 mM/1, and the chloride is present at a concentration of about 118.0 to 136.5 mM/1, or the electrolyte components can be diluted to create hypotonic formulations where the ratios between the electrolyte concentrations remain unchanged.

The ophthalmologically suitable compositions can further optionally include calcium, magnesium and phosphate. In such embodiments, in isotonic preparations, the calcium is preferably present at a concentration of about 0.5 to 2.0 mM/1, the magnesium is preferably present at a concentration of about 0.3 to 1. 1 mM/1, and the phosphate is preferably present at a concentration of about 0.8 to 2.2 mM/1. For hypotonic formulations, the electrolyte components can be diluted to create hypotonic formulations where the ratios between the electrolyte concentrations remain unchanged.

Accordingly, particular embodiments the invention are set forth in the Examples. The final pH of the ophthalmologically suitable compositions generally ranges from about 7.0 to 8. 0, as measured by, for example, a Fisher pH Accumet Model 600. However, this pH range need not be rigidly adhered to, and it may be desirable to alter pH outside of this range, for instance, to improve ophthalmic drug penetration through the ocular surface. In view of the teachings provided herein, those skilled in the art may employ other pH ranges.

The ophthalmologically suitable compositions of the invention can be applied to the ocular surface by various methods known in the art. For example, the compositions can be applied topically to the ocular surface as eye drops or ointments. The preparation can also be applied using an eye cup so that the eye is bathed. The compositions can also be applied using a continuous or near continuous infusion device for ocular surface irrigation and/or wetting and/or drug delivery. The compositions can also be applied by release from a sustained-release carrier such as a sustained-release polymer, a liposome or a microcapsule. The compositions may also be applied by devices that spray solutions as required onto the surface of the eye. The invention is further directed to methods of using the compositions described above to treat a subject, e.g., a subject having or at risk of having an infection, e.g., an infection of the eye or skin. The method comprises the step of applying the ophthalmologically suitable compositions described herein to the site of the infection, or site where an infection is likely to occur, or the site from which an infection might originate, for a time and under conditions effective for reducing the amount of bacteria present. In a specific embodiment, the time and conditions selected result in an at least about 1 log reduction in colony-forming units of the infecting bacteria after one minute of exposure to the antimicrobial preparation. In other embodiments, the application of the ophthalmologically suitable composition for one minute results in an at least about 2, 3, 4 or 5 log reduction in colony-forming units.

The invention also provides methods of treating ocular disorders such as blepharitis, dry eye, eye inflammatory disorders, infectious conjunctivitis, and other ocular disorders that result from or are complicated by bacterial colonization or infection of the eye or surrounding tissue, by applying the ophthalmologically suitable compositions provided herein to the eye and/or surrounding tissue of a subject.

The invention also provides methods of treating infection of the ocular surface by applying the ophthalmologically suitable compositions provided herein to the eye of a subject. Exemplary infections that can be treated with the antimicrobial preparations provided herein include conjunctivitis, e.g., infectious conjunctivitis and corneal ulcers.

The invention also provides methods of preventing an eye infection in a subject having an eye surgery or procedure. These methods would comprise applying an ophthalmologically suitable composition described herein to the eye over a number of days preceding the surgery or procedure to reduce or eliminate the risk of developing an infection during the surgery or procedure. Exemplary procedures include cataract or LASIK surgery. The invention also provides methods of maintaining low bacterial colony counts on punctal plugs that have been placed in patients for treatment. Exemplary punctal plugs include those manufactured by Odyssey Medical (Memphis, TN), and Eagle Vision (Memphis, TN).

Application of the ophthalmologically suitable compositions set forth herein can be by any one of a number of art recognized methods. For example, application can be by an applicator, such as a Qtip or pad, by drops from a dropper or bottle, or using a finger or fingers.

The ophthalmologically suitable compositions of the invention may be applied one or more times per day, and may be left in place as long as needed, depending on the intended indication. The number of days which a subject applies the ophthalmologically suitable composition, and the duration of the application, will depend on the intent of treatment or on the location and severity infection, and efficacy of the preparations on a given infection. The ophthalmologically suitable composition may be applied for a period of 30 seconds, 45 seconds, 1 minute, 2 minutes, 3 minutes, 4 minutes, 5 minutes, or longer. The ophthalmologically suitable composition can be applied by release from a sustained release carrier such as a sustained-release polymer, a liposome or a microcapsule. The ordinary skilled physician would be able to effectively prescribe a treatment regimen that will be effective in treating or preventing an infection in an individual.

The methods described above may further include a rinsing step after a recommended period of exposure. This step preferably comprises a simple water rinse. The ophthalmologically suitable composition may be rinsed from the area to which it was applied with ample water after application, e.g., with a hand, finger or any moist pad or cloth suitable for this purpose.

In further embodiments, the invention provides disinfecting a surface. Exemplary surfaces include those on medical instruments, in medical facilities, on medical devices, and those in a home, e.g., in a kitchen or bathroom.

Commercial Applications

The methods and compositions of the invention find numerous commercial applications that could beneficially utilize compliance enhancing methods and compositions for antibacterial applications. Consequently, the invention includes a kit comprising the compositions of the invention, e.g., a kit for the treatment of an infection, e.g., an ocular infection, an ocular disorder, eyelid hygiene. The kits optionally include an applicator. The composition can be in the form of drops, solution, paste, cream, foam, gel, or ointment, or the like, when included in the kits of the invention.

The kit may optionally be packaged with instructions for use. The kit may optionally contain a dispenser or applicator, e.g., a sponge, to apply the ophthalmologically suitable compositions of the invention to the infected area of a subject.

EXAMPLES

A study was conducted to evaluate the stability and efficacy of exemplary compositions of the eye care preparations described herein.

Example 1 : Formulation and Efficacy of Compositions Containing Activated Chlorine Dioxide

EQUIPMENT

325=/-2.5°C Incubator, VWR S/N 1000590 22.5±/-2.5°C Incubator, Forma S/N 32889120 Spectrophotometer, Spectronic 2ODT, S/N 3DU9337006 Centrifuge, ID 34440967 pH Meter, ID 0157189 Osmometer, ID T09390 MATERIALS

1. Staphylococcus aureus, ATCC 6538

2. Pseudomonas aeruginosa, ATCC 9027

3. Escherichia coil, ATCC 8739

4. Candida albicans, A TCC 10231 5. Aspergillus niger, ATCC 16404

6. Soybean Casein Digest Agar, (SODA)

7. Sabouraud Dextrose Agar, (SDA)

8. D/E Neutralizing Broth, (DEB)

9. 0.1N Sodium Thiosulfate Volumetric Solution 10. Potassium Iodide

11. Starch Indicator Solution

12. 2.5N Hydrochloric Acid

EXPERIMENTS After compounding the compositions, samples were submitted for the initial

USP Antimicrobial Effectiveness testing per USP and SOP-OOl 81. All test samples were challenged with approximately 1.0x10 to 1.0x10⁶ cfu/mT, of S. aureus ATCC 6538, P. aeruginosa ATCC 9027, E, coli ATCC 8739, C. albicans, ATCC 10231 and A. niger, ATCC 16404. The organisms were inoculated into a 5 OmL centrifuge tube containing 1OmL of test sample at Time=0. One (1) mL was aliquoted from each centrifuge tube for the following 4 weeks. The log reduction was determined by the plate count method after 7 14, 21 and 28 days by diluting in DEB from 10 ' to 10^"4 for bacteria/yeast and 10^"l to 10^" for mold. The plates were then poured with the appropriate media and incubated. Bacterial plates were poured with SCDA and incubated at 32.5±².5°C. Yeast/mold plates were poured with SCDA and incubated at 22.5±2.5°C. The initial antimicrobial effectiveness data was reviewed before any further testing for antimicrobial effectiveness was conducted. Only formulas that had met the acceptance criteria at the initial testing were followed beyond the initial testing.

The acceptance criteria for USP Antimicrobial Effectiveness for a Category 1 item are:

Bacteria - 7 day - Not less than 1.0 log reduction from initial count

14 day - Not less than 3.0 log reduction from initial count

10 21 day - No specification

28 day - No increase from 14 days count at 28 days Yeast/Molds - 7 and 14 days - No increase from initial count

21 days - No specification

28 days - No increase from initial count

15 (No increase is defined as NMT 0.5 logio)

RESULTS

Cl

Constituent Target % Target (mg/mL)

Sodium Carboxymethylcellulose, 7H3 SXF-PH 0.2500 2.500

Sodium Chloride, USP 0.18663 1.8663

Potassium Chloride, USP 0.06145 0.6145

Magnesium Chloride, Hexahydrate, USP 0.00420 0.0420

Sodium Phosphate, Monobasic, Monohydrate, USP 0.00471 0.0471

Calcium Chloride, Dihydrate, USP 0.00541 0.0541

Sodium Bicarbonate, USP 0.09231 0.9231

Boric Acid,NF 0.1500 1.50

Sodium Borate, Decahydrate, USP 0.00008 0.0008

Chlorine Dioxide, USP 0.0050 0.0500

Osmo = 132 pH=7.22 0

Osmo= 148 pH=7.39 Results

The three solutions exhibited above are at 0.25% CMC and 0.005% Chlorine Dioxide.

Electrolytes and buffers are adjusted appropriately to obtain various osmolalities between 130 and 200 mθsm/kg.

All three solutions pass USP Antimicrobial Effectiveness Testing. Sample Description

Sample 1 - Cl Sample 2 - C2 Sample 3 - C3

Criteria - Bacteria - 7 day - not less than 1.0 log reduction from initial count 14 day - not less than 3.0 log reduction from initial count 21 day - no specification 28 day - no increase from 14 days count at 28 days

Yeast/Molds - 7 and 14 days - no increase from initial count 21 days - no specification 28 days - no increase from initial count (No increase is defined as nmt 0.5 logi₀)

The pH of solution C3 was adjusted to 7.22 with 0.51g of IN HCl.

Formulations are effective with or without boric acid and/or hydrochloric acid for pH adjustment.

Solution C3 had an osmolality of 132 mOsm/kg. It was prepared and aliquoted into 2 dram LDPE bottles. These bottles were placed at 40°C/NMT 25% RH for 3 months. The pH, osmolality, viscosity and chlorine dioxide testing was performed at 3 month timepoint. Sample C3 was tested at 2.5 months for Preservative Efficacy. Table 1 shows the results from the testing for pH, osmolality, viscosity and Chlorine Dioxide. Table 2 shows the results from the Preservative Efficacy testing.

Table 2 : Preservative E icac results a ter 2.5 months accelerated stabilit )

Example 2: Formulation and Efficacy of Compositions Containing Activated Chlorine Dioxide and Trehalose An exemplary composition of the invention is described in this example and data demonstrating the efficacy of this composition is also provided.

Preparation of Composition C 4 Phase I

A solution was made by measuring 800.2 g of water into a 1000 mL bottle. A stir bar was added to the solution and the beaker was placed on a hot plate. The solution was heated to 70 °C and 5.02 g of Sodium Carboxymethylcellulose was slowly added. The solution was mixed on a hot plate for 30 minutes, removed from heat and allowed to cool to room temperature to dissolve the CMC. The solution was diluted to 1000.1 g with water. The solution was autoclaved at 121 °C for 50 minutes. This solution was termed "Phase I".

Phase II

The solution was made by measuring 413.98 g of water into a 600 mL beaker.

To this solution 0.3778 g of Sodium Chloride, 0.1246 g Potassium Chloride, 0.0082 g of Magnesium Chloride, Hexahydrate, 0.0096 g of Sodium Phosphate, Monobasic, Monohydrate, 0.01 12 g of Calcium Chloride, Dihydrate, 0.1872 g of Sodium Bicarbonate, 1.5012 g of Boric Acid, 0.0008 g of Sodium Borate, Decahydrate, 34.1997 g of Trehalose and 2.00 g Chlorine Dioxide (2.5%) were added and stirred until dissolved. The solution was diluted to 500.04 grams with purified water. This solution was termed "Phase II".

Phase III

The solution was prepared by mixing 450.03 g of Phase I and 450.06 g of Phase II in a 1000 mL bottle. The pH of the solution was found to be 7.44.

C4 was tested for the ability to kill S. aureus, P. aeruginosa, E. coli, C. albicans, and A. Niger. As indicated in Table 3 below, C4 effectively kills all of the microorganisms.

Table 3

Example 3: Formulation of Compositions Containing 1% CMC exemplary gel composition was prepared by increasing the CMC content hown below for composition C5.

Example 4: Formulation of Compositions Containing Dyphilline

An exemplary composition was prepared by including dyphilline as shown below for composition C6.

Example 5 : Efficacy of the Compositions of Examples 3 and 4

C5 and C6 were tested for their ability to kill S. aureus, P. aeruginosa, E. coli, C. albicans, and A. Niger as descirbed above. As indicated in Table 4 below, C5 and C6 effectively kill all of the microorganisms.

Table 4

Incorporation by Reference

The contents of all references, patents, pending patent applications and published patents, cited throughout this application are hereby expressly incorporated by reference.

Equivalents

Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by the following claims.

Claims

What is claimed is:

1. An ophthalmologically suitable composition comprising activated chlorine dioxide, wherein the osmolality of the composition is less than aboutl 80 mOsm/Kg.

2. The ophthalmologically suitable composition of claim 1, wherein the chlorine dioxide is present in a concentration of about 10-125 ppm.

3. The ophthalmologically suitable composition of claim 2, wherein the chlorine dioxide is present in a concentration of about 25-75 ppm.

4. The ophthalmologically suitable composition of claim 3, wherein the chlorine dioxide is present in a concentration of about 50 ppm.

5. The ophthalmologically suitable composition of claim 1 , further comprising boric acid.

6. The ophthalmologically suitable composition of claim 1 , further comprising a buffer component having a functional group selected from the group consisting of borate, phosphate, carbonate, and combinations thereof.

7. The ophthalmologically suitable composition of claim 1, wherein the final pH of the composition is about 6.5 to about 8.

8. The ophthalmologically suitable composition of claim 7, wherein the final pH of the preparation is about 7.2.

9. The ophthalmologically suitable composition of claim 1, further comprising trehalose.

10. The ophthalmologically suitable composition of claim 1, further comprising dyphilline.

1 1. The ophthalmologically suitable composition of claim 1, wherein the composition is in the form of a solution, cream, paste, ointment, or gel.

12. The ophthalmologically suitable composition of claim 1, wherein the composition is a solution.

13. The ophthalmologically suitable composition of claim 25, wherein the activated preparation has an osmolality of about 160 mOsm/Kg or less.

14. The ophthalmologically suitable composition of claim 1 , wherein the ophthalmologically suitable composition has an osmolality of about 180 mOsm/Kg or less and wherein the chlorine dioxide is present in a concentration of about 50-75 ppm.

15. The ophthalmologically suitable composition of claim 1, further comprising carboxymethylcellulose.

16. The ophthalmologically suitable composition of claim 14, wherein the carboxymethylcellulose is sodium carboxymethylcellulose.

17. The ophthalmologically suitable composition of claim 15, wherein the sodium carboxymethylcellulose present at a final concentration of about 0.25%.

18. The ophthalmologically suitable composition of claim 17, wherein the sodium carboxymethylcellulose present at a final concentration of about 1.0%.

19. The ophthalmologically suitable composition of claim 1 , wherein the ophthalmologically suitable composition is for application to the eye, eye-lid, eye margin, punctal plug or a contact lens.

20. The ophthalmologically suitable composition of claim 1 , wherein the composition does not contain dyphilline.

21. The ophthalmologically suitable composition of claim 1 , wherein the composition is in the form of a gel.

22. An ophthalmologically suitable composition comprising the following components at about the following final concentrations:

Sodium Carboxymethylcellulose 0.2500%

Sodium Chloride 0.03777%

Potassium Chloride 0.01244%

Magnesium Chloride, Hexahydrate, 0.00085%

Sodium Phosphate, Monobasic, 0.00095%

Monohydrate

Calcium Chloride, Dihydrate 0.00110%

Sodium Bicarbonate 0.01868%

Boric Acid, NF 0.15000%

Sodium Borate, Decahydrate 0.00008%

Chlorine Dioxide 0.00500%, and

Trehalose 3.420%.

23. The ophthalmologically suitable composition of claim 22, wherein the pH is adjusted to about 7.2.

24. The ophthalmologically suitable composition of claim 22, wherein the composition has an osmolality of about 145-155 mOsm/Kg.

25. A method of treating a microbial colonization or infection in or around the eye of a subject comprising;

applying the ophthalmologically suitable composition of claim 1 to the colonized or infected area in an amount sufficient to treat a microbial colonization or infection in a subject.

26. The method of claim 25, wherein the microbial colonization or infection is bacterial, fungal or yeast colonization or infection.

27. The method of claim 25, wherein the ophthalmologically suitable composition is in the form of a drop, solution, cream, paste, ointment, or gel.

28. A kit comprising the ophthalmologically suitable composition of claim 1 and instructions for use.