US 20070249546A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0249546A1 Sawaya (43) Pub. Date: Oct. 25, 2007

(54) OPHTHALMIC AND RELATED AQUEOUS Publication Classification SOLUTIONS CONTAINING ANTIFUNGAL AGENTS, USES THEREFOR AND METHODS (51) Int. Cl. FOR PREPARING THEMI A61K 3 1/7048 (2006.01) A6II 3L/496 (2006.01) (76) Inventor: Assad S. Sawaya, Baiting Hollow, NY A61K 31/4196 (2006.01) (US) A61K 31/4178 (2006.01) (52) U.S. Cl...... 514/28: 514/254.07: 514/383; Correspondence Address: 514/397: 514/772.7 DARBY & DARBY P.C. P.O. BOX 770 Church Street Station (57) ABSTRACT New York, NY 10008-0770 (US) The invention relates generally to concentrates and aqueous (21) Appl. No.: 11/738,451 Solutions for topical application comprising antifungal addi tives or agents as well as to preparation and use of Such (22) Filed: Apr. 20, 2007 concentrates and solutions. More specifically, the invention Related U.S. Application Data relates to preparation and use of Solutions that come in contact with the eyelids and/or eyes, such as but not limited (60) Provisional application No. 60/794.240, filed on Apr. to contact lens Solutions, aqueous ophthalmic rinse solu 22, 2006. tions, and aqueous Surgical scrubs for ophthalmic use. Patent Application Publication Oct. 25, 2007 Sheet 1 of 4 US 2007/0249546 A1

eTest Control - Fusarium Solani o 0.02%TONAFTATE 104 4-hours (100)

a 0.02%TOLNAFATE a 0.02%TOLNAFATE 6-hours (100) 24-hours (100) Patent Application Publication Oct. 25, 2007 Sheet 2 of 4 US 2007/0249546 A1

Fig-6

a 0.02%MCONOZOE e O,O296 MCONOZOE 4-hours (100) 6-hours (100)

Fig-7

• O.O2%MCONOZOLE 24-hours (100) Patent Application Publication Oct. 25, 2007 Sheet 3 of 4 US 2007/0249546 A1

Fig. 8A OSNFECNG EFFECACY EST 83000 ATHE-1

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2 O O O O 3600 4300 O 5 1o 15 2O 25 30 EXPOSURE TIME (Hour)

Fig. 8B OSNFECTING EFFCACY EST 83000 ATHE-2

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Fig. 8C OSNFECTING EFFCACY TEST 90000 ATHE-3 83000

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O 5 O 15 2O 25 3O EXPOSURE TIME (Hour) Patent Application Publication Oct. 25, 2007 Sheet 4 of 4 US 2007/0249546 A1

Fig. 9 DSNFECTING EFFICACY TEST (FUSARIUM SOLAN) 6

5 A.9

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OPHTHALMIC AND RELATED AQUEOUS fungal agents could not be used and were not contemplated SOLUTIONS CONTAINING ANTIFUNGAL for use in aqueous solution preparations for ocular use AGENTS, USES THEREFOR AND METHODS FOR because they were inadequately soluble or even dispersible. PREPARING THEM 0006 Conventional ophthalmic solutions and pre-surgi cal eye lid scrubs suffer from similar deficiencies vis a vis REFERENCE TO PROVISIONAL APPLICATION protection against fungal contamination. Such solutions and 0001) This application claims the benefit of U.S. Provi scrubs are Vulnerable to similar fungal contamination, which sional Application No. 60/794,240, filed on Apr. 22, 2006. leaves Subjects using the Solution and/or scrubs Vulnerable to fungal infection. As a result of Such contamination, FIELD OF THE INVENTION Subjects using conventional multipurpose lens solutions 0002 The invention relates generally to concentrates and and/or conventional ophthalmic aqueous Solutions and pre aqueous solutions for topical application comprising anti Surgical eye lid scrubs may develop conditions such as fungal additives or agents as well as to preparation and use Fusarium keratitis eye infections and/or Fusarium conjunc of Such concentrates and solutions. More specifically, the tivitis. Some infections may result to damage to the cornea. invention relates to preparation and use of Solutions that In some cases, corneal transplants are required, and on some come in contact with the eyelids and/or eyes, such as but not occasions the Subjects suffer loss of sight. limited to contact lens solutions, aqueous ophthalmic rinse 0007. The use point, the eye, precludes organic solvents, Solutions, and aqueous Surgical Scrubs for ophthalmic use. such as alcohols, because they are irritant or toxic. While conventional multipurpose lens Solutions contain preserva BACKGROUND OF THE INVENTION tive systems, recent events have demonstrated that Such Solutions are still Vulnerable to contamination by fungal 0003. In conventional antifungal preparations for oph species such as Fusarium Solani, which leaves the Subject thalmic or periophthalmic use, the antifungal agent is sus using conventional Solutions vulnerable to fungal infection." pended in ointments, creams and non-aqueous Solutions of See FDA News, Apr. 10, 2006 reporting the presence of Fusarium in oil-based delivery systems. Conventional antifungal agents multipurpose contact lends solutions manufactured by Bausch & Lomb precipitate when introduced into an aqueous solution, which resulting in 109 cases of Fusarium keratitis. See FDA Update regarding Contact Lenses and Eye Infections dated Apr. 13, 2006 and FDA Statement diminishes or extinguishes their antifungal properties, ren dated Apr. 14, 2006 regarding voluntary recall of Bausch & Lomb Contact dering Such agents impractical for use in aqueous solutions Lens Solution. such as, but not limited to, multipurpose lens solutions. 0008 Heretofore no one has come forward with a method 0004 The prior art lacks methods of preparing oph for providing an effective amount of an antifungal agent in thalmic preparations that are homogenous aqueous solutions an aqueous ophthalmic or periophthalmic solution, wherein comprising antifungal agents for use in and around the eye. the solution is physiologically acceptable for ophthalmic or Instead, the prior art provides ophthalmic preparations com periophthalmic use. Nor has a preparation containing an prising antifungal agents in the form of Suspensions. For antifungal agent in solution in a suitable physiologically example, U.S. Pat. No. 7,056,893 (hereafter the 893 patent) acceptable vehicle been devised such that it can be mixed discloses using a polymeric Suspending agent like polyeth with an aqueous ophthalmic or periophthalmic solution and ylene glycol to prepare an aqueous polymeric Suspension, impart to Such a solution antifungal properties without However, a skilled worker will appreciate that a polymeric precipitating the antifungal agent. Thus, a need for a safe Suspension is not the equivalent of a clear, homogenous antifungal preparation formulated for delivery in the form of aqueous solution, especially for use with the eye. Published a solution that may be used in aqueous Solutions, such as U.S. patent application No. US2004/0198829 discloses the multipurpose lens Solutions and ophthalmic, topical and addition of prostanoids to a suspension comprising a thera Surgical Solutions, is readily apparent. The antifungal prepa peutic agent like an antifungal Such as Voriconazole to rations in the form of an aqueous solution presented herein effectively increase transport and/or penetration of the thera will be effective and safe to reduce the risk of various fungal peutic agent in the eye. However, neither the 893 patent nor infections inherent in the currently available multipurpose US2004/0198829 provides a homogenous ophthalmic or lens Solutions, and ophthalmic, topical and pre-surgical periophthalmic aqueous Solution comprising an effective solutions, which lack the benefit of the technology presented amount of an antifungal agent. herein. The antifungal preparations in the form of an aque 0005 Published U.S. patent application No. US2004/ ous solution presented herein may also increase the shelflife 0266702, another example in the prior art, describes an of conventional ophthalmic Solutions and pre-surgical eye ophthalmic composition that uses a polymeric Suspension lid scrubs. agent such as a carboxy-containing polymer Such as poly mers of acrylic acid to deliver to the eye an azalide antibiotic BRIEF SUMMARY OF THE INVENTION alone or in combination with another agent like an antifun 0009. In accordance with this invention, there is provided gal agent such as miconazole nitrate. US2004/0266702 a method for preparing a concentrate solution containing an discloses that the polymeric Suspension agent may be poly antifungal agent in a form Suitable for Subsequent introduc ethylene glycol and may additionally comprise a surfactant. tion in an aqueous medium Suitable for ophthalmic or The preparations disclosed in US2004/0266702 are aqueous periophthalmic use, the method comprising the steps of: a) polymeric suspensions. US2004/0266702 is silent as to a dissolving an antifungal agent in a physiologically accept Suitable method comprising steps for obtaining an aqueous able non-aqueous vehicle to form a non-aqueous solution of Solution comprising an azalide antibiotic and/or an antifun said antifungal agent said vehicle being miscible with aque gal agent without precipitation of the azalide antibiotic or ous solutions; b) adding a physiologically acceptable Sur antifungal agent. Thus, prior to the current invention, anti factant to said non-aqueous solution in an amount Sufficient US 2007/0249546 A1 Oct. 25, 2007 to stabilize said antifungal agent upon its Subsequent intro contact of the eye of said Subject with an aqueous solution duction in said aqueous medium; and c) mixing the non during ophthalmic or periophthalmic use of said aqueous aqueous Solution and Surfactant mixture until a clear con Solution, the method comprising: providing as the aqueous centrate solution is obtained. Solution used ophthalmically or periophthalmically an aque 0010. The present invention also provides a method for ous solution suitable for ophthalmic or periophthalmic use to inhibiting fungal growth in an aqueous Solution Suitable for which has been added a concentrate comprising (a) an ophthalmic or periophthalmic use comprising the step of antifungal agent dissolved in a non-aqueous vehicle, said dissolving in said aqueous solution a concentrate Solution vehicle being miscible with an aqueous solution suitable for containing an antifungal agent dissolved in a physiologically ophthalmic or periophthalmic use containing no antifungal acceptable non-aqueous vehicle, said vehicle being miscible agent, and (b) one or more physiologically acceptable Sur with said aqueous solution and one or more physiologically factants, said concentrate having been added in an amount acceptable surfactants, said concentrate solution being Sufficient to provide a fungicidally or fungistatically effec added to said aqueous Solution in an amount Sufficient to tive amount of said antifungal agent in said provided aque provide a fungicidally or fungistatically effective amount of ous solution. said antifungal agent in said aqueous Solution. BRIEF DESCRIPTION OF DRAWINGS 0011. The invention provides a concentrate solution con taining an antifungal agent in a form Suitable for Subsequent 0016 FIGS. 1-7 are photographs of plates showing the introduction in an aqueous medium Suitable for ophthalmic growth of Fusarium solani in the absence (FIG. 1) or or periophthalmic use, said concentrate Solution made by: a) presence (FIGS. 2-7) of aqueous solutions comprising anti dissolving an antifungal agent in a physiologically accept fungally effective amounts of tolnaftate or miconazole able non-aqueous vehicle to form a non-aqueous solution of nitrate. said antifungal agent said vehicle being miscible with aque 0017 FIG. 8 is a graph of data collected from a disin ous solutions; b) adding a physiologically acceptable Sur fecting efficacy test using antifungal triplex hydrated solu factant to said non-aqueous solution in an amount Sufficient tions (ATHE-1 (FIG. 8A), ATHE-2 (FIG. 8B) and ATHE-3 to stabilize said antifungal agent upon its Subsequent intro (FIG. 8C)) to reduce the growth of Fusarium solani. FIG. 9 duction in said aqueous medium; and c) mixing the non is a graph of the growth reduction data from the disinfecting aqueous Solution and Surfactant mixture until a clear con efficacy test shown as log reductions. centrate solution is obtained. 0012. The invention also provides an aqueous solution DETAILED DESCRIPTION OF THE for ophthalmic or periophthalmic use comprising an anti INVENTION fungal agent in an amount effective to inhibit fungal growth 0018. As used herein and in the appended claims, the in said aqueous solution, said solution made by dissolving in singular forms a “an', and “the include plural referents a first aqueous solution containing no antifungal agent a unless the context clearly dictates otherwise. Thus, for second concentrate Solution containing (a) an antifungal example, reference to “an antifungal agent' includes a agent dissolved in a physiologically acceptable non-aqueous plurality of such agents known to those skilled in the art, and vehicle, said vehicle being miscible with said first aqueous so forth. Solution containing no antifungal agent; and (b) one or more physiologically acceptable Surfactants; said concentrate hav 0019. Unless defined otherwise, all technical and scien ing been added to the first aqueous Solution containing no tific terms used herein have the same meaning, as commonly antifungal agent in an amount Sufficient to provide a fungi understood to one of ordinary skill in the art to which this cidally or fungistatically effective amount of said antifungal invention belongs. agent in the combined solution. 0020. The term “antifungal agent” refers to a substance 0013 In some embodiments, the invention provides a capable of imparting fungistatic and/or fungicidal proper concentrate Solution for ophthalmic or periophthalmic use ties. containing from about 0.005% (w/w) to about 20.0% (w/w) 0021. The term “solution” refers to one or more solutes antifungal agent, from about 20% (w/w) to about 80% (w/w) that are completely dissolved or dispersed in a solvent non-aqueous vehicle, and from about 20% (w/w) to about yielding a homogenous mixture. 70.0% (w/w) surfactant. 0022. In some embodiments, the subject is a human. In 0014. In other embodiments, the invention provides an Some embodiments, the Subject is a veterinary Subject or an aqueous solution for ophthalmic or periophthalmic use com experimental animal, e.g., a rodent. prising from about 0.001% (w/w) to about 2.0% (w/w) of an antifungal agent Such as miconazole nitrate or tolnaftate, I. Aqueous Solutions Comprising Antifungal Agents and from about 0.5% (w/w) to about 2.0% (w/w) of a vehicle Methods of Preparing the Same such as polyethylene glycol 400, from about 0.3% (w/w) to 0023 Aqueous ophthalmic solutions containing antifun about 10.0% (w/w) of a surfactant such as polysorbate 80 gal agents avoid the risk of various fungal infections inher and, optionally, from about 0.3% (w/w) to about 10.0% ent in currently available solutions, such as, but not limited (w/w) of another surfactant such as octoxynol 40. In other to, multipurpose lens solutions, and ophthalmic, topical and embodiments, the aqueous Solutions of the invention may pre-surgical solutions, because these conventional Solutions additionally comprise additional agents such as polyduater do not contain antifungal agents. The present invention nium-1, polyhexamethylene biguanide. provides aqueous solutions comprising effective amounts of 0.015 The invention also provides a method for avoiding antifungal agents that remain in solution and impart anti fungal infection in a Subject, said infection arising from fungal properties to the Solutions while maintaining these US 2007/0249546 A1 Oct. 25, 2007

Solutions as physiologically acceptable (e.g., nonirritant and antifungal properties to a topical aqueous solution of oph nontoxic). The aqueous solutions are homogenous and clear. thalmic or periophthalmic use. In another embodiment, the The aqueous solutions comprising effecting amounts of aqueous Solution comprising an antifungal agent is func antifungal agents are not suspensions. The present invention tionally stable for at least the minimum reasonable shelf life provides aqueous Solutions for ophthalmic or perioph of Such products. thalmic use comprising physiologically acceptable, yet 0028 Antifungal agents which can be used herein are effective, amounts of antifungal agents. physiologically acceptable at the effective amounts 0024. While not intending to be bound by any theory, it employed and, particularly Suitable for ophthalmic or is believed that the described antifungal concentrates and periophthalmic uses. Antifungal agents for use in the anti aqueous Solutions comprising antifungal agents maintain the fungal concentrates and/or topical aqueous solutions com antifungal agents in Solution with low to no precipitation by prising effective amounts of antifungal agents include, but use of a non-aqueous, but water Soluble, physiologically are not limited to amorolfine, amphotericin B, anidulafun acceptable solvent for the antifungal agent (e.g., polyethyl gin, butoconazole, butenafine, caspofungin, ciclopiroX ola ene glycol) and one or more stabilizing agents like a mine, clotrimazole, econazole, fluconazole, flucytosine, physiologically acceptable surface active agent (e.g., a Sur griseofulvin, haloprogin, itraconazole, ketoconazole, micaf factant). It also is believed that the antifungal properties are ungin, miconazole (including miconazole nitrate), naftifine, maintained in the aqueous solution comprising an effective nikkomycin Z. nystatin (topical and liposomal), oxicona amount of antifungal agents because the antifungal agents Zole, posaconazole, pimaricin, ravuconazole, Sulconazole, Substantially remain in Solution. terbinafine, terconazole, tioconazole, tolnaftate, unde cylenate, Voriconazole, or any other antifungal agent or a 0.025 A. Antifungal Concentrate salt thereof known to those of skill in the art may be used. 0026. The present invention employs an antifungal con The antifungal agents used in the invention include free acid, centrate comprising an antifungal agent, a physiologically free base, salts and esters. In one embodiment, the antifungal acceptable non-aqueous vehicle in which the agent is agent is tolnaftate. In one embodiment, the antifungal agent soluble, and a physiologically acceptable Surfactant. In one is miconazole nitrate. embodiment, the invention provides a method for preparing 0029. In all embodiments of the antifungal concentrate, a concentrate comprising the steps of: 1) dissolving one or the antifungal agents may be used in effective concentrations more antifungal agents in a physiologically acceptable non generally ranging from about 0.005% (w/w) to about 6.0% aqueous vehicle; 2) adding one or more physiologically (w/w). Higher concentrations are permitted subject to the acceptable Surfactants; and 3) mixing (e.g., by stirring) until amounts of physiological acceptability, but are not neces a clear Solution is obtained. In one embodiment, the inven sary. In one embodiment of the antifungal concentrate, an tion provides a method of preparing a concentrate compris antifungal agent may be used in concentrations ranging from ing the steps of: 1) applying heat (within the range of 50° about 0.01% (w/w) to about 4.0% (w/w). In one embodiment C.-85°C. that is sufficient to dissolve an antifungal agent of the antifungal concentrate, the antifungal agent may be into a physiologically acceptable non-aqueous vehicle with used in a concentration ranging from about 0.06% (w/w) to out destroying the antifungal properties of the antifungal about 2.0% (w/w). In one embodiment of the antifungal agent; 2) adding a physiologically acceptable Surfactant concentrate, miconazole nitrate may be used at a concen (within the temperature range of 30° C.-40° C. with con tration of 0.1% (w/w) or 2.0% (w/w). In one embodiment, tinual mixing); and 3) mixing until a clear Surfactant and tolnaftate may be used at a concentration of 0.1% (w/w) or antifungal containing Solution is obtained. The mixing and 2.0% (w/w). dissolving steps can be combined. In a preferred embodi ment of the method of preparing the antifungal concentrate 0030. In all embodiments of the antifungal topical aque of the invention, the antifungal agent is dissolved into the ous Solution, antifungal agents may be used in effective physiologically acceptable non-aqueous vehicle without concentrations generally ranging from about 0.001% (w/w) destroying the antifungal properties of the antifungal agent to about 2.0% (w/w). Higher concentrations are permitted Subject to the amounts of physiological acceptability, but are while applying heat within the range of 75° C.-80° C. not necessary. In one embodiment of the antifungal topical 0027. In another embodiment, the invention provides a aqueous solution, an antifungal agent may be used in con method of preparing an aqueous solution Suitable for oph centrations ranging from about 0.005% (w/w) to about thalmic or periophthalmic use that comprises an effective 0.05% (w/w). In one embodiment of the antifungal topical amount of an antifungal agent dissolved therein. An effective aqueous Solution, the antifungal agent may be used in a amount of the antifungal agent remains in Solution imparting concentration ranging from about 0.01% (w/w) to about 0. antifungal properties to the solution throughout the useful 1% (w/w). In one embodiment of the antifungal topical life of the Solution. In one embodiment, an antifungal aqueous solution, miconazole nitrate may be used at a concentrate of the invention is added to a topical aqueous concentration of 0.01% (w/w) or 0.02% (w/w). In one Solution to form an aqueous solution having antifungal embodiment of the antifungal topical aqueous solution, properties. In another embodiment, the antifungal concen tolnaftate may be used at a concentration of 0.01% (w/w) or trate is added to a topical aqueous Solution. Preferably, an 0.02% (w/w). The solubility of each antifungal agent used in effective amount of the antifungal agent remains in Solution the current invention in a physiologically acceptable non for the useful life of the solution. The useful shelf life of the aqueous vehicle can be determined using methods well antifungal concentrate or an aqueous solution comprising known in the art. Additionally, for each antifungal agent the antifungal concentrate is storage-stable at room tempera used in the current invention specific determinations can be ture of at least about 24 months or greater. In another conducted to determine the concentration of Surfactant Suf embodiment, the invention provides a method of imparting ficient to aid in dissolution of the antifungal agent. US 2007/0249546 A1 Oct. 25, 2007

0031. In all embodiments, the antifungal agent used is the art may be used. In one embodiment, the vehicle or nontoxic and nonirritant (i.e. does not cause ocular irritation solvent is polyethylene glycol 400. of the lens or conjunctiva of the eye) when administered to a subject. In all embodiments, the antifungal agent is used at 0034) To aid in dissolution of the concentrates including a concentration that is safe for administration to a subject. In the antifungal agent(s) dissolved therein in an aqueous all embodiments, the concentration of the antifungal agent Solution, the present invention employs Surface-active agents (i.e. Surfactants). In all embodiments, the Surfac used by a method of the invention is nontoxic for the tant(s) used herein is physiologically acceptable and in duration of administration to a subject. preparation of a topical aqueous solution comprising an 0032) Physiologically acceptable non-aqueous vehicles antifungal agent. In some embodiments, the physiologically that may be used to dissolve an antifungal(s) include, acceptable Surfactants are Suitable for ophthalmic and without limitation: polyols, polyhydric alcohol polymers, periophthalmic uses. In all embodiments, the Surfactant(s) polyhydric alcohol ethers, polyhydric alcohol esters. The used in the present invention include, but are not limited to, non-aqueous vehicles are soluble in water. In some embodi nonionic and cationic Surfactants. Anionic Surfactants ments, the polyhydric alcohol ether is polyethylene glycol. should be avoided. In some embodiments, the polyethylene glycol has an aver age molecular weight generally in the range from about 4 to 0035. Nonionic or neutral surfactants that may be used about 160,000. In other embodiments, the polyethylene herein include polysorbates and polyethoxylates. In some glycol has an average molecular weight generally in the embodiments, nonionic Surfactants to be used herein range from about 8 to about 1000. In other embodiments, the include, but are not limited to, polysorbate 20, polysorbate polyethylene glycol has an average molecular weight gen 40, polysorbate 60, polysorbate 61, polysorbate 65, polysor erally in the range from about 100 to about 600. In other bate 81, polysorbate 85, polysorbate 80, polysorbate 80 embodiments, the polyethylene glycol has an average acetate, octoxynol 1, Octoxynol 3, octoxynol 5, octoxynol 6, molecular weight generally in the range from about 60 to octoxynol 7, octoxynol 8, octoxynol 9, octoxynol 10, octox about 100. ynol 11, octoxynol 12, octoxynol 13, Octoxynol 16, octox 0033. Vehicles or solvents which may be used herein ynol 25, octoxynol 30, octoxynol 33, octoxynol 40, octox include, but are not limited to, polyethylene glycol 4, ynol 70, octoxynol-9 carboxylic acid, octoxynol-20 polyethylene glycol 6, polyethylene glycol 7, polyethylene carboxylic acid, or any other Surfactant Suitable for oph glycol 8, polyethylene glycol 9, polyethylene glycol 10, thalmic and/or dermatologic purposes known to those of polyethylene glycol 12, polyethylene glycol 14, polyethyl skill in the art may be used. ene glycol 16, polyethylene glycol 18, polyethylene glycol 20, polyethylene glycol 32, polyethylene glycol 40, poly 0036 Suitable surfactants to be used herein include cat ethylene glycol 45, polyethylene glycol 55, polyethylene ionic Surfactants such as quaternary ammonium compounds. glycol 60, polyethylene glycol 75, polyethylene glycol 90, In another embodiment, cationic Surfactants used herein are, polyethylene glycol 100, polyethylene glycol 135, polyeth but are not limited to, benzalkonium chloride and benzalko ylene glycol 150, polyethylene glycol 180, polyethylene nium saccharinate, or any other Surfactant Suitable for oph glycol 200, polyethylene glycol 220, polyethylene glycol thalmic or periopthalmic purposes known to those of skill in 240, polyethylene glycol 350, polyethylene glycol 400, the art may be used. polyethylene glycol 500, polyethylene glycol 600, polyeth 0037. In some embodiments of the antifungal concen ylene glycol 800, polyethylene glycol 2000, polyethylene trate, Surfactants may be used in concentrations ranging glycol 5000, polyethylene glycol 7000, polyethylene glycol from about 20% (w/w) to about 70% (w/w). In some 9000, polyethylene glycol 14000, polyethylene glycol embodiments of the antifungal topical aqueous solution, 20000, polyethylene glycol 23000, polyethylene glycol Surfactants may be used in concentrations ranging from 45000, polyethylene glycol 90000, polyethylene glycol about 0.3% (w/w) to about 10.0% (w/w). In a preferred 115M, polyethylene glycol 160M, propylene glycol, propy embodiment of the antifungal topical aqueous solution, lene glycol alginate, propylene glycol behenate, propylene Surfactants may be used in concentrations ranging from glycol butyl ether, propylene glycol capreth-4, propylene about 0.5% (w/w) to about 2% (w/w). In another preferred glycol caprylate, propylene glycol ceteth-3 acetate, propy embodiment of the antifungal topical aqueous solution, lene glycol ceteth-3 propionate, propylene glycol citrate, surfactants may be used at a concentration of about 1.0% propylene glycol coate, propylene glycol dicaprate, propy (w/w). In all embodiments, the Surfactant used is nontoxic lene glycol dicaproate, propylene glycol dicaprylate, propy and nonirritant when administered to a Subject. In all lene glycol dicocoate, propylene glycol diethylhexanoate, embodiments, the Surfactant is used at a concentration that propylene glycol diisononanoate, propylene glycol diisos is safe for administration to a subject. In one embodiment, tearate, propylene glycol dilaurate, propylene glycol the surfactant is a combination of polysorbate 80 and dioleate, propylene glycol dipelargonate, propylene glycol octoxynol 40. In another embodiment, the surfactant is distearate, propylene glycol hydroxy Stearate, propylene gly polysorbate 80. In another embodiment, the polysorbate 80 col isoceteth-3 acetate, propylene glycol isostearate, propy in the antifungal concentrate is used in a concentration of lene glycol laurate, propylene glycol laureth-6, propylene 48% (w/w). In another embodiment, the polysorbate 80 in glycol linolenate, propylene glycol myristate, propylene the antifungal topical aqueous solution is used in a concen glycol myristyl ether, propylene glycol myristyl ether tration of 1.0% (w/w). In another embodiment, the surfac acetate, propylene glycol oleate, propylene glycol oleate SE, tant is octoxynol 40. In another embodiment, octoxynol 40 propylene glycol oleth-5, propylene glycol propyl ether, in the antifungal concentrate is used at a concentration of propylene glycol stearate, propylene glycol Stearate SE, or 50% (w/w). In another embodiment, the octoxynol 40 in the any other vehicles or solvents suitable in ophthalmic solu antifungal topical aqueous solution is used in a concentra tions as described herein that are known to those of skill in tion of 2.0% (w/w). In one embodiment, the combination of US 2007/0249546 A1 Oct. 25, 2007

polysorbate 80 and octoxynol 40 in the antifungal topical nents of said tonicity agents and buffers are nontoxic, and do aqueous solution is used in a concentration of 1.0% (w/w). not distort the vision of the Subject using the antifungal 0038 B. Aqueous Solutions for Combination with the concentrate. Suitable tonicifiers that may be added to the Antifungal Concentrates antifungal concentrates and/or aqueous solutions comprising antifungal agents include, without limitation, dextrose, 0039. One aspect of the present invention is to add the potassium chloride and/or sodium chloride. Suitable buffers antifungal concentrate described herein to a first aqueous that may be added to the antifungal concentrate and/or Solution containing no antifungal agent to yield an aqueous aqueous Solution comprising antifungal agents include, Solution imparting antifungal properties to said first aqueous Solution without precipitating the antifungal agent. Another without limitation, boric acid, Sodium borate, Sodium or aspect of the present invention provides an aqueous solution potassium citrate, Sodium bicarbonate, sodium phosphate, for ophthalmic or periophthalmic use comprising an anti and potassium phosphate. The buffers may be used in fungal agent in an amount effective to inhibit fungal growth concentrates ranging from about 0.3% (w/w) to about 3.5% in the aqueous solution that is made by dissolving in a first (w/w). aqueous Solution containing no antifungal agent a second 0043. Additionally, to also guard against bacterial infec antifungal concentrate Solution described herein. Aqueous tion, antibacterial agents found in conventional ophthalmic solutions provided by the present invention for opthalmic or Solutions such as multipurpose lens solutions may be added periophthalmic use comprising an antifungal concentrate to the antifungal concentrates and/or aqueous solutions described herein are homogenous. First aqueous solutions comprising antifungal agents. In this regard, the antibacterial that may be employed to add to the antifungal concentrate of agents are nontoxic, and do not distort the vision of the the invention include, but are not limited to, aqueous solu Subject using the antifungal concentrate or aqueous solution tions which come into contact with the eyelids and/or eyes, comprising the same. Antibacterial agents for use in the Such as multipurpose lens solutions, ophthalmic rinse solu antifungal concentrates and/or aqueous solutions comprising tions, Surgical Scrubs for eye use, eye drops, eye wash antifungal agents include, but are not limited to, polyami Solutions, contact lens solutions, topical over the counter nopropyl biguanide, alexidine hydrochloride, polyduater ocular and periocular solutions (i.e., artificial tears), ocular nium, polyguaternium 42, myristamidopropyl dimethy and periocular cleaning solutions, eye irrigating Solutions, lamine, or other Suitable agents known to a skilled worker. and/or antibacterial Solutions for Surgical scrubs or topical In some embodiments, polyaminopropyl biguanide may be application. In other words, the first aqueous Solution may used in concentrations ranging from 0.0001% (w/w) to contain one or more physiologically acceptable active ingre 0.002% (w.fw) in the antifungal concentrate and/or aqueous dients such as the foregoing and/or excipients. Solution comprising antifungal agents. In other embodi 0040. In some embodiments, an antifungal concentrate of ments, alexidine hydrochloride may be used in concentra the invention may be added to a commercially available tions ranging from about 0.0002% (w/w) to about 0.006% contact lens Solution or a multipurpose lens solution to (w/w) in the antifungal concentrate and/or aqueous solution impart antifungal properties without precipitation of the comprising antifungal agents. antifungal agent. In some embodiments, an antifungal con 0044) In some embodiments, the aqueous solutions com centrate of the invention may be added to an aqueous prising antifungal agents also comprise a comfort or mois Solution prepared for use as a contact lens or multipurpose turizing agent to provide hydration and lubrication of the lens solution that is not commercially available to impart lens. Such agents include, but are not limited to, polyduater antifungal properties without precipitation of the antifungal. nium 10, poloxamer, propylene glycol, hydroxypropylmeth 0041. In some embodiments, an antifungal concentrate of ylcellulose (HPMC), or other suitable agents known to a the invention is added to an ocular or periocular cleaning skilled worker. In all embodiments, the cleaning agents are Solution to impartantifungal properties without precipitation nontoxic, and do not distort the vision of the Subject using of the antifungal agent. In some embodiments, the cleaning the antifungal concentrate or aqueous Solution comprising Solution comprises cleaning agents to effectively clean a lens the same. In some embodiments, a comfort or moisturizing of film deposits and Surface debris. Such cleaning agents that agent may be used in concentrations ranging from 0.01% may be used include, without limitation, poloxamers and (w/w) to 2.0% (w/w) in the antifungal concentrate and/or tetronic Surfactants comprising poly(oxyethylene) hydro aqueous solution comprising antifungal agents. In a pre philic units. In some embodiments, a topical aqueous solu ferred embodiment, a comfort or moisturizing agent may be tion that has been added to an antifungal concentrate of the used in concentrations ranging from 0.1% (w/w) to 0.8% invention additionally comprises one or more of the follow (w/w) in the antifungal concentrate and/or aqueous solution ing cleaning agents in concentrations ranging from about comprising antifungal agents. 0.0001% (w/w) to 1.0% (w/w): Pluronic(R) F-127 (poloxam 0045 Since, in some embodiments, the aqueous solutions ine), Tetronic R. 904, Tetronic R 304, or other suitable clean comprising antifungal agents are intended to be adminis ing agents. In all embodiments, the cleaning agents are tered topically to the eyelids and/or eye, it is preferred that nontoxic, and do not distort the vision of the Subject using they be free of pathogenic organisms and/or sterile. Abenefit the antifungal concentrate or aqueous solution comprising of a sterile solution is that it reduces the possibility of the same. introducing contaminants into a subject when the antifungal 0042. In other embodiments, an antifungal concentrate of concentrates and/or aqueous Solutions comprising antifungal the invention may be added to tonicity agents and buffers agents of the present invention are administered topically, that are found in conventional ophthalmic and perioph for example, to the eyelids and/or eye. Sterility or adequate thalmic solutions to impart antifungal properties without antimicrobial preservation may be provided as part of the precipitation of the antifungal. In this regard, the compo present antifungal preparations. In some embodiments, the US 2007/0249546 A1 Oct. 25, 2007 antifungal concentrates and/or aqueous solutions comprising antifungal agents of the invention are, but not limited to, eye antifungal agents of the present invention are produced conditions with a fungal-based etiology. In some embodi under sterile conditions. ments, the Fusarium Solani is responsible for the fungal 0046. In lieu of or additional to sterilization, the aqueous based etiology. In some embodiments, the antifungal con Solutions comprising antifungal agents may contain a physi centrates and/or aqueous solutions comprising antifungal ologically acceptable preservative to minimize the possibil agents made by the methods of the invention may be used in ity of microbial contamination. A physiologically acceptable a Subject for the prevention of a fungal-based eye condition. preservative may be used in the present antifungal prepara 0050. In other embodiments, fungal infections for which tions to increase the stability of the antifungal preparations. concentrates or aqueous solutions made by methods of the Preservatives suitable for use herein include, but are not current invention may have activity against include, but are limited to, polyaminopropylbiguanide, polyhexamethylene not limited to: Fusarium keratitis, Fusarium conjunctivitis, biguanide (PHMB), polyduaternium-1, myristamidopropyl. and endophthalmitis. In one embodiment, the fungal infec and Sorbic acid. In some embodiments, preservatives may be tion to be prevented or to be avoided is Fusarium keratitis. used in concentrations ranging from about 0.00005% (w/w) In one embodiment, the antifungal concentrates and/or aque to about 0.00015% (w/w). In one embodiment, polyduate ous Solutions comprising antifungal agents made by the mium-1 may be used in concentrations ranging from about methods of the invention are administered for prevention of 0.001% (w/w) to about 0.002% (w/w). In another embodi Fusarium keratitis. In another embodiment, the fungal infec ment, myristamidopropyl (w/w) may be used at a concen tion to be prevented or to be avoided is Fusarium conjunc tration of about 0.0005% (w/w). In another embodiment, tivitis. In one embodiment, the antifungal concentrates and/ Sorbic acid may be used in concentrations ranging from or aqueous solutions comprising antifungal agents made by about 0.1% (w/w) to about 0.3% (w/w). the methods of the invention is administered for prevention II. Uses for Aqueous Solutions Containing Antifungal of Fusarium conjunctivitis. Agents 0051. The antifungal aqueous solutions of the invention 0047 Subjects who use the antifungal concentrates and/ comprising antifungal agents may be administered topically or aqueous solutions comprising antifungal agents produced or may be used, e.g., on contact lenses during storage. Most by methods of the present invention may attain previously commonly, the Solutions of the invention can be applied as unavailable levels of protection against fungal organisms eye drops, eye washes, eye lid Scrubs and the like. and species. As a result, Subjects using such antifungal concentrates and/or aqueous solutions comprising antifungal 0.052 In order that this invention may be better under agents produced by methods of the present invention may stood, the following examples are set forth. These examples evade Vulnerability to exposure to fungal organisms and are for the purposes of illustration only and are not to be may prevent or may avoid the development of fungal construed as limiting the scope of this invention in any infections caused by fungal genera. The antifungal concen a. trates and/or aqueous solutions comprising antifungal agents made by methods of the invention are intended for admin EXAMPLE I istration to a Subject to prevent or to avoid a fungal infection. 0053 An aqueous topical solution comprising antifungal 0.048 Fungal genera for which the antifungal concen agents was produced by a method of the invention as trates and/or aqueous solutions comprising antifungal agents follows: made by methods of the current invention may have activity against include, but are not limited to Fusarium, Penicil 0054 A. Antifungal Concentrate lium, Aspergillus, Cephalosporium (Acremonium), Tricho 0055 An antifungal concentrate was prepared by dissolv phyton, Microsporum, Epidermophyton, Scopulariopsis, and ing 0.2 grams miconazole nitrate in 10.0 grams polyethylene Candida. In some embodiments, a fungal genus against glycol (PEG) 400 with continual mixing at a temperature of which the antifungal concentrates and/or aqueous solutions 75°-85° C. until the miconazole nitrate was completely comprising antifungal agents may be used is Fusarium. In solubilized. This non-aqueous Solution was then cooled to Some embodiments, fungal species for which the present 30° -40° C. Next, with continual stirring, 5.6 grams of invention may have activity against include, but are not Tween R 80 (polysorbate 80) was added followed by addi limited to: Fusarium solari, Fusarium Solani, Fusarium tion of 4.5 grams of octoxynol 40 to stabilize the concen avenaceum, Fusarium culmorum, Fusarium balbigenum, trate. Fusarium caeruleum, Fusarium conglutinans, Fusarium lini, Fusarium oxysporum, Fusarium vasinfectum, Fusarium 0056 B. Multipurpose Lens Solution graminearum, Trichophyton crateriform, Trichophyton 0057. An aqueous multipurpose lens solution was pre rubrum, Trichophyton mentagpophytes, Trichophyton inter pared by adding 0.25 grams of disodium ethylenediamine digitalis, Trichophyton verrucosum, Trichophyton megnini, tetraacetic acid (EDTA), 0.60 grams of sodium borate, 5.0 Trichophyton gallinae, Trichophyton sulphureum, Tricho grams of boric acid, 9.0 grams of propylene glycol, 1.0 phyton Schoenleini, Microsporum audonini, Microsporum grams of Pluronic RF-127, 0.6 grams of polycuaternium 10 canis, Microsporum gypsum, Epidermophytonfloccosum, and 1.8 grams of sodium chloride to 859.35 grams of Scopulariopsis brevicaulis, and combinations thereof. In purified water. Some embodiments, a fungal species against which the antifungal concentrates and/or aqueous solutions comprising 0.058 C. Cleaning Solution antifungal agents may be used is Fusarium solari. 0059 A cleaning solution was prepared by adding, while 0049 Fungal infections that may be avoided with anti continually stirring, 0.5 grams of Tetronic R. 904, 0.1 grams fungal concentrates and/or aqueous solutions comprising of Tetronic R 304 and 1.5 grams of polyaminopropyl bigu US 2007/0249546 A1 Oct. 25, 2007

anide to 100 grams of purified water. (Surfactants sold under the 06.1360B multipurpose lens solution containing 0.1% the trademark Tetronic are tetrafunctional block copolymer (w/w) polyduatemium-1 with continual mixing at 35° C. nonionic Surfactants terminating in primary hydroxyl until a clear solution was obtained. Lot 061360B was cooled groups.) to room temperature. Lot 061360B is a clear, aqueous solution which contains 0.02% (w/w) Miconazole Nitrate, 0060 A final aqueous solution was prepared by mixing 1.6% (w/w) PEG 400, 0.5% (w/w) Tween(R) 80 (polysorbate the antifungal concentrate A. with the multipurpose lens 80), 0.45% (w/w) octoxynol 40, and 0.001% (w/w) Solution B to form a clear homogenous solution. The result polyduaternium-1. ing aqueous Solution was filter sterilized using a 0.22 micron filter. Another final aqueous solution also was prepared by EXAMPLE IV mixing the antifungal concentrate A. with the cleaning Solution C. to form a clear, homogenous solution. The 0068 Aqueous topical solutions comprising antifungal resulting aqueous solution was filter sterilized using a 0.22 agents prepared according to the methods of the present micron filter. invention were used to test their fungicidal properties against Fusarium Solani over a period of 24 hours. EXAMPLE II Preparation of Fusarium Solani Culture Suspension 0061 Aqueous topical solution 061360A (Lot 061360A) 0069. A Fusarium solani culture suspension was pre comprising 0.02% (W/w) miconazole nitrate was prepared as pared as follows: From a working slant a culture of follows: Fusarium Solani was transferred to the Surface area of a 0062. A 061360 antifungal concentrate was prepared by Roux bottle containing 200 mL of Sabourand Dextrose Agar. dissolving 0.2 grams miconazole nitrate in 16.0 grams PEG The Roux bottle and its contents were incubated at 20-25°C. 400 with continual mixing at a temperature of 75°-80° C. for 3 to 7 days. Next, the surface area of the Roux bottle was until the miconazole nitrate was completely solubilized. The rinsed with 50 mL of sterile saline solution, which subse solution was cooled to 35° C.t5° C. Next, with continual quently was centrifuged at no more than 4000 xg for a stirring, 5.0 grams of Tween R 80 (polysorbate 80) was maximum of 15 minutes. To determine the culture popula tion, serial dilutions were performed using Sabourand Dex added followed by addition of 4.5 grams of octoxynol 40 to trose Agar in a ratio of 1 to 10 at a minimum of 1x107 to stabilize the concentrate. 1 x10 dilution. Each dilution was plated and incubated at 0063 A 061360A aqueous multipurpose lens solution 20-25° C. for 3 to 7 days. The number of Fusarium solani containing 0.1% PHMB was prepared by adding 0.25 gram cfu per dilution was determined by counting the culture of EDTA, 0.60 gram of sodium borate, 4.0 grams of boric growth on each plate. acid, 9.0 grams of propylene glycol, 1.0 grams of Poloxamer 407, 0.5 gram of Tetronic R. 904, 0.1 gram of Tetronic.R. 304, Treatment of Fusarium Solani with Aqueous Solutions 0.6 gram of PHMB and 1.0 gram of sodium chloride one Comprising 0.02% (w/w) Tolnaftate or 0.02% (w/w) ingredient at a time to 956.35 grams of purified water with Miconazole Nitrate continual mixing and heating to 70° -80° C. After dissolu 0070 Inoculums of Fusarium solani culture solution that tion of all ingredients, the 061360A aqueous multipurpose were used to conduct the experiments for which data is lens solution containing 0.1% PHMB was then cooled to shown in FIGS. 1-7 was prepared by diluting 0.1 mL of a room temperature. 1.66x10 cfu/mL Fusarium solani culture suspension into 10 0064. Lot 061360A was prepared by slowly adding the mL of aqueous solutions comprising 0.02% (W/w) tolnaftate 061360 antifungal concentrate to the 061360A multipurpose or 0.02% (w/w) miconazole nitrate providing a final count of lens solution containing 0.1% PHMB with continual mixing 1.66x10" cfu/mL (typical range of 1x10" to 1x10 cfu/mL). at 35° C. until a clear solution was obtained. Lot 061360A The inoculated sample was stored at 20-25° C. was cooled to room temperature. Lot 061360A is a clear, 0071 Viable counts of inoculated samples were deter aqueous solution which contains 0.02% Miconazole Nitrate, mined at 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, and 24 1.6% PEG 400, 0.5% Tween(R) 80 (polysorbate 80), 0.45% hours by removing 1.0 mL aliquots of the inoculated sample octoxynol 40, and 1 ppm PHMB. were used. To each 1.0 mL aliquot, 9 mL of phosphate buffered saline (PBS) containing 0.5 % Tween R 80 EXAMPLE III (polysorbate 80) was added to perform serial decimal dilu tions. The viable count of Fusarium Solani was determined 0065 Aqueous topical solution 06.1360B (Lot 061360B) by dilution into culture plates using recovery medium (Sab comprising 0.02% (W/w) miconazole nitrate was prepared as ourand Dextrose Agar). The plates were incubated at 20-25° follows: C. for 3 to 7 days. The number of Fusarium solani cfu was 0066. A 061360B aqueous multipurpose lens solution determined by counting the culture on the plates. Log containing 0.1% polyduaternium-1 was prepared by adding reduction of Fusarium Solani cfu was calculated for each 0.25 gram of EDTA, 0.60 gram of sodium borate, 4.0 grams specific time point. of boric acid, 9.0 grams of propylene glycol, 1.0 gram of 0072. As discussed above, plates containing an initial Poloxamer 407, 0.5 gram of Tetronic R904, 0.1 gram of inoculum comprising 1.66x10" cfu Fusarium solani were Tetronic R304, 12.0 grams of polyduatemium-1 and 1.0 monitored over a period of 24 hours in the absence (See FIG. gram of sodium chloride one ingredient at a time to 945.85 1) or presence (See FIGS. 2-7) of aqueous solutions com grams of purified water with continual mixing and heating to prising 0.02% (w/w) tolnaftate or 0.02% (w/w) miconazole 70°-80° C. After dissolution of all ingredients, the 06.1360B nitrate, as prepared by methods of the invention. All plates aqueous multipurpose lens solution containing 0.1% that were treated with the aqueous solutions comprising polyduaternium-1 was then cooled to room temperature. 0.02% (w/w) tolnaftate or 0.02% (w/w) miconazole nitrate 0067 Lot 061360B was prepared by slowly adding the for a period of 4, 6 or 24 hours showed reduced growth of 061360 antifungal concentrate (described in Example II) to Fusarium Solani. FIG. 2 is a photograph of a plate showing US 2007/0249546 A1 Oct. 25, 2007

30 cfu remaining after 4 hours treatment with 0.02% (w/w) aqueous solution to 35°C.-5°C.; 3) with continual stirring, tolnaftate, which represents a 3.39 log reduction in the 27% (w/w) Tween R 80 was added; 4) then with continual growth of Fusarium Solani. FIG. 3 is a photograph of a plate stirring at a temperature of 35° C.t5° C., 36% at 35° C.i.5° showing 30 cfu remaining after 6 hours treatment with C., 23% (w/w) Oxtoxynol 40 was added; 5) the non-aqueous 0.02% (w/w) tolnaftate, which shows a 3.46 log reduction in solution was cooled to 25° C. +3° C.; and 6) the resulting the growth of Fusarium Solani. FIG. 4 is a photograph of a tolnaftate concentrate was added with continual mixing at a plate showing 10 cfu remaining 24 hours after treatment temperature of 25°C.-3°C. to purified water. ATHE-2 is a with 0.02% (w/w) tolnaftate, which represents a 3.94 log clear, aqueous solution for ophthalmic and periophthalmic reduction in the growth of Fusarium solani. FIG. 5 is a use that contains 0.02% tolnaftate, 1% PEG 400, 0.56% photograph of a plate showing 0 (zero) cfu remaining 4 Tween R. 80, and 0.46% octoxynol 40. hours after treatment with 0.02% (w/w) miconazole nitrate, which represents a 4.91 log reduction in the growth of 0076) The solution ATHE-3 was prepared by 1) dissolv Fusarium Solani. FIG. 6 is a photograph of a plate showing ing 0.99% (w/w) miconazole nitrate in 49% (w/w) PEG 400 0 (zero) cfu remaining 6 hours after treatment with 0.02% with continual mixing at a temperature of 75°C.-80°C. until (w/w) miconazole nitrate, which represents a 4.91 log reduc the miconazole nitrate was completely solubilized; 2) cool tion in the growth of Fusarium solani. FIG. 7 is a photo ing the non-aqueous solution to 35° C.t5° C.; 3) with graph of a plate showing 0 (zero) cfu remaining 24 hours continual stirring at a temperature of 35° C.-5°C., 27% after treatment with 0.02% (w/w) miconazole nitrate, which (w/w) Tween R 80 was added; 4) then continual stirring at a represents a 4.91 log reduction in the growth of Fusarium temperature of 35° C.t5° C., 23% (w/w) oxtoxynol 40 was Solani. added; 5) the non-aqueous solution was cooled to 25°C.3° C.; and 6) the resulting miconazole nitrate concentrate was EXAMPLE V added with continual mixing at a temperature of 25° C.3° C. to purified water. ATHE-3 is a clear, aqueous solution for 0.073 Aqueous solutions comprising the following anti ophthalmic and periophthalmic use that contains 0.02% fungal agents: 0.01% (w/w) tolnaftate (ATHE-1); 0.02% miconazole nitrate, 1% (w/w) PEG 400, 0.55% Tween(R) 80, (w/w) tolnaftate (ATHE-2); or 0.02% (w/w) miconazole and 0.46% octoxynol 40. nitrate (ATHE-3) were prepared according to methods of the invention and were tested for their fungicidal properties 0077. The aqueous solution comprising 0.02% (w/w) against Fusarium Solani in a disinfectant efficacy test. tolnaftate was added to an initial inoculum of 83,000 cfu per mL of Fusarium Solani, and after four (4) hours of exposure 0074 The solution ATHE-1 was prepared by 1) dissolv time, the initial inoculum was reduced to 37 cfu (See Table ing 0.66% (w.fw) tolna?tate in 33% (w.fw) PEG 400 with 1 below and FIG. 8). This result demonstrates a log reduc continual mixing at a temperature of 75° C.-80° C. until the tion of 3.39 in Fusarium solani growth after four (4) hours tolnaftate was completely solubilized; 2) cooling the non of exposure time (See Table 1 below and FIG. 9). The aqueous solution to 35°C.-5°C.; 3) with continual stirring aqueous solution comprising 0.01% (w/w) tolnaftate was at a temperature of 35° C.i.5° C., 36% (w/w) Tween R 80 added to an initial inoculum of 83,000 cfu per mL of was added; 4) then with continual stirring 35°C.-5°C., 30% Fusarium Solani, and after four (4) hours of exposure time, (w/w) octoxynol 40 was added; 5) the homogenous non the initial inoculum was reduced to 3,600 cfu (See Table 1 aqueous solution was cooled to 25° C.3° C. and 6) the below and FIG. 8). This result demonstrates a log reduction resulting tolnaftate concentrate was added with continual of 1.29 in Fusarium solani growth after four (4) hours of mixing at a temperature of 25°C.-3°C. to purified water. exposure time (See Table 1 below and FIG. 9). The aqueous ATHE-1 is a clear, aqueous solution for ophthalmic and solution comprising 0.02% (w/w) miconazole nitrate was periophthalmic use that contains 0.01% tolnaftate, 0.5% added to an initial inoculum of 83,000 cfu per mL of PEG 400, 0.55% Tween R. 80, and 0.46% octoxynol 40. Fusarium Solani, and after four (4) hours of exposure time, 0075) The solution ATHE-2 was prepared by 1) dissolv the initial inoculum was reduced to zero (0) cfu (See Table ing 0.99% (w/w) tolna?tate in 49% (w/w) PEG 400 with 1 below and FIG. 8). This result demonstrates a log reduc continual mixing at a temperature of 75° C.-80° C. until the tion of 4.91 in Fusarium solani growth after four (4) hours tolnaftate was completely solubilized; 2) cooling the non of exposure time (See Table 1 below and FIG. 9).

TABLE 1. Disinfectant efficacy test results for ATHE-1, ATHE-2, and ATHE-3 aqueous solutions Exposure F solani Plate Log Concentration Ingredients Time Countml Saline Control Reduction ATHE-2 4 hours 37 x 100 Initial Inoculum 3.39 (0.02% Tolna?tate; 1% PEG 3 x 10 Population 400; 0.56% Tween (R) 80: 0 x 102 84, 82 x 10' 0.46% Octoxynol 40) 1 x 10 10, 8 x 10 0 x 10 0.1 ml inoculated into 0 x 10 10 ml of ATHE-2 solution Inoculum population 8.3 x 10' per ml 6 hours 28 x 109 Initial Inoculum 3.46 3 x 10 Population 0 x 102 84, 82 x 10' 0 x 10 10, 8 x 10 US 2007/0249546 A1 Oct. 25, 2007

TABLE 1-continued Disinfectant efficacy test results for ATHE-1, ATHE-2, and ATHE-3 aqueous solutions Exposure F solani Plate Log Concentration Ingredients Time Coun ml Saline Control Reduction Ox 04 0.1 ml inoculated into Ox Os Om of ATHE-2 Solution OCl. lum population 8.3 x 10 per ml 24 hours 9 x initia Inoculum 3.94

0, 8 0.1 ml inoculated into Om of ATHE-2 Solution OCl. lum population 8.3 x 10 per ml ATHE-1 4 hours >80 x initia Inoculum 1.29 (0.01% Tolna?tate: 0.5% PEG 400; 0.55% Tween (R) 80; 0.46% Octoxynol 40) 0.1 ml inoculated into Om of ATHE-1 Soluti Oil noculum population 8.3 x 10 per ml 6 hours >80 x initia Inoculum 1.30

0.1 ml inoculated into 0 ml of ATHE-1 Soluti Oil noculum population 8.3 x 10 per ml 24 hours >80 x initial Inoculum 1.39 Population

0.1 ml inoculated into of ATHE-1 ion noculum population 10 per ml (ATHE-3) 4 hours Ox Inoculum 4.91 (0.02% Miconozole Nitrate; ation 1% PEG 400; 0.55% (w/w) Tween (R) 80; 0.46% 0, 8 octoxynol 40) 0.1 ml inoculated into Om of ATHE-3 Soluti Oil noculum population 8.3 x 10 per ml 6 hours Ox initia Inoculum 4.91 Popu ation 84, 82 x 10' 0, 8 x 10 0.1 ml inoculated into Om of ATHE-3 Soluti Oil noculum population 8.3 x 10 per ml 24 hours Ox initia Inoculum 4.91 Popu ation 84, 82 x 10' 0, 8 x 10 0.1 ml inoculated into Om of ATHE-3 Soluti Oil noculum population 8.3 x 10 per ml US 2007/0249546 A1 Oct. 25, 2007 10

EXAMPLE VI form a cup into which the test solution was dropped. Each 0078 Ocular Irritation tests (Standard Operating Proce eyelid was then gently held together for approximately one dure (SOP) 16G-45, Ocular Irritation Test (ISO)) were second to prevent loss of the test solution. The left eye of conducted (Pacific BioLabs, Hercules Calif., USA) for aque each rabbit served as an untreated control. Administration of ous solutions of the present invention comprising 0.01% the test solution, the eyes were examined 1, 24, 48, and 72 (w/w)-0.2% (w/w) miconazole nitrate. The tests were con hours after treatment. At each observation period, the ocular ducted to determine the potential irritating properties of reaction to treatment was graded according to a numerical Solutions of aqueous solutions comprising the antifungal scoring system for evidence of corneal ulceration or opacity, concentrate of the invention by assessment in an eye irrita inflammation of the iris, or redness, chemosis, and any tion test in three white female New Zealand rabbits weigh discharge of the conjunctivae (Biological Evaluation of ing between 2.5 to 2.9 kg. The aqueous Solutions, which were prepared according to the present invention, Lot Medical Devices-Part 10: Tests for Sensitization and Irrita 061360A (preparation is described in Example II) and Lot tion. ISO 10993-10:2002(E)). All grades are based upon on 061360B (preparation is described in Example III) were a scale increasing in range from 0-4, depending upon the tested. 0.1 mL of the test solution, either Lot 061360A or Lot specific criteria being observed. All animals remained 061360B, was placed into the right eye of each rabbit. healthy throughout the study period. No irritation in vivo Administration of the test solution was completed by gently was observed with Lot 061306A (See Table 2) or Lot pulling each lower eyelid away from the rabbit eyeball to 061306B (See Table 3) throughout the study period. TABLE 2

Ocular Irritation Scores for Lot 061360A

Cornea Coniunctivae

Percent Red Animal Wit. Time after Opacity Area Iris ness - Chenosis Discharge Number Sex (kg) Dosing (hrs) L. R L R L. R. R. L. L R. L. R 4.0833 F 2.7 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O 40871 F 2.6 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O 40874 F 2.5 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O R = Right Eye L = Left Eye

0079 TABLE 3

Ocular Irritation Scores for Lot 06136OB

Cornea Coniunctivae

Percent Red Animal Wit. Time after Opacity Area Iris ness - Chenosis Discharge Number Sex (kg) Dosing (hrs) L. R L R L. R. R. L. L R. L. R 40830 F 2.7 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O 40831 F 2.7 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O 40832 F 2.9 1 O O O O O O O O O O O O 24 O O O O O O O O O O O O 48 O O O O O O O O O O O O 72 O O O O O O O O O O O O R = Right Eye L = Left Eye US 2007/0249546 A1 Oct. 25, 2007

EXAMPLE VII c) mixing the non-aqueous solution and surfactant mix ture until a clear concentrate solution is obtained. 0080 Cytotoxity studies were also conducted for aque 2. The method of claim 1, wherein the antifungal agent is ous solutions comprising the antifungal concentrate of the added in a concentration within the range of 0.005% (w/w) invention. The cytotoxity test was conducted (Pacific to 6.0% (w/w). BioLabs, Hercules Calif., USA; SOP 15A-02 and SOP 3. The method of claim 1, wherein the surfactant is added 15B-01) using test materials, Lot 061360A (preparation is in a concentration within the range from 0.5% (w/w) to 2.0% described in Example II) and Lot 061360B (preparation is (w/w). described in Example III), in the United States Pharmaco 4. The method of claim 1, wherein said vehicle is poly poeia (USP) standard minimum essential media (MEM) ethylene glycol having a molecular weight within the range elution test against L-929 mouse fibroblast cells (ATCC Cell from 8 to 1000. Line CCL1, NCTC Clone 929). Test article extract was 5. The method of claim 1, wherein said surfactant is prepared using 3.0 grams of test material consisting of a polysorbate 80. representative portion of the test article that was extracted in 6. The method of claim 1, wherein said antifungal agent 15.0 mL of 5% serum supplemented MEM at 37° C. t1 °C. is selected from the group consisting of amorolfine, ampho in a humidified incubator with 5%+1% CO for not less than tericin B, anidulafungin, butoconazole, butenafine, caspo 24 hours. Positive and negative controls were also extracted fungin, ciclopirox olamine, clotrimazole, econazole, flu under the same conditions. After the 24 hour extraction conazole, flucytosine, griseofulvin, haloprogin, period, the growth medium from duplicate 10 cm wells itraconazole, ketoconazole, micafungin, miconazole, each containing the monolayer of L-929 cells were decanted miconazole nitrate, naftifine, nikkomycin Z, topical nystatin, and replaced with 2 mL of test article extract. The control liposomal nystatin, oxiconazole, posaconazole, pimaricin, extracts were tested in the same manner as the extracting ravuconazole, sulconazole, terbinafine, terconazole, tio medium containing test article. All cell cultures were incu conazole, tolnaftate, undecylenate, and Voriconazole. bated for 48+2 hours at 37° C.1° C. in a humidified 7. A method for inhibiting fungal growth in an aqueous incubator with 5%+1% CO. The cells were scored for solution suitable for ophthalmic or periophthalmic use com cytotoxic response at 48 hours. The Lot 061360A and Lot prising the step of dissolving in said aqueous solution a 061360B aqueous solutions (test materials) each scored a 0 concentrate solution containing an antifungal agent dis (Zero) indicating no biological reactivity (i.e. no cytotoxic solved in a physiologically acceptable non-aqueous vehicle, ity). Possible biological reactivity scores were 0 (zero) said vehicle being miscible with said aqueous solution and indicating no reactivity, 1 indicating Slight Reactivity, 2 one or more physiologically acceptable surfactants, said indicating Mild Reactivity, 3 indicating Moderate Reactiv concentrate solution being added to said aqueous solution in ity, and 4 indicating Severe Reactivity. According to USP an amount sufficient to provide a fungicidally or fungistati specifications, a test material meets the requirements of the cally effective amount of said antifungal agent in said test if the cell culture treated with the test material does not aqueous solution. score greater than a grade of 2 (Mild Reactivity). The test 8. The method of claim 7, wherein said vehicle is poly results are presented in Table 4 for Lot 061360A and Lot ethylene glycol having a molecular weight within the range 061360B aqueous solutions (test materials) prepared accord from 8 to 1000. ing to the present invention. 9. The method of claim 7, wherein said antifungal agent is selected from the group consisting of amorolfine, ampho TABLE 4 tericin B, anidulafungin, butoconazole, butenafine, caspo fungin, ciclopirox olamine, clotrimazole, econazole, flu Cytotoxicity Scores conazole, flucytosine, griseofulvin, haloprogin, Test Material Grade Biological Reactivity itraconazole, ketoconazole, micafungin, miconazole. 6.1360A O None miconazole nitrate, naftifine, nikkomycin Z, topical nystatin, O6136OB O None liposomal nystatin, oxiconazole, posaconazole, pimaricin, Positive Control Extract 4 Severe ravuconazole, sulconazole, terbinafine, terconazole, tio Negative Control Extract O None conazole, tolnaftate, undecylenate, and Voriconazole. 10. A concentrate solution containing an antifungal agent in a form suitable for subsequent introduction in an aqueous medium suitable for ophthalmic or periophthalmic use, said We claim: concentrate solution made by: 1. A method for preparing a concentrate solution contain a) dissolving an antifungal agent in a physiologically ing an antifungal agent in a form suitable for subsequent acceptable non-aqueous vehicle to form a non-aqueous introduction in an aqueous medium suitable for ophthalmic solution of said antifungal agent said vehicle being or periophthalmic use, the method comprising the steps of: miscible with aqueous solutions: b) adding a physiologically acceptable surfactant to said a) dissolving an antifungal agent in a physiologically non-aqueous solution in an amount sufficient to stabi acceptable non-aqueous vehicle to form a non-aqueous lize said antifungal agent upon its subsequent introduc solution of said antifungal agent said vehicle being tion in said aqueous medium; and miscible with aqueous solutions: c) mixing the non-aqueous solution and surfactant mix b) adding a physiologically acceptable surfactant to said ture until a clear concentrate solution is obtained. non-aqueous solution in an amount sufficient to stabi 11. The concentrate of claim 10, wherein the antifungal lize said antifungal agent upon its subsequent introduc agent is added in a concentration within the range of 0.005% tion in said aqueous medium; and (w/w) to 6.0% (w/w). US 2007/0249546 A1 Oct. 25, 2007

12. The concentrate of claim 10, wherein the surfactant is c) adding 27.5% (w/w) polysorbate 80 and 22.1% (w/w) added in a concentration within the range from 0.5% (w/w) Octoxynol 40 to said nonaqueous solution in an amount to 2.0 (w/w). Sufficient to stabilize said miconZole nitrate upon its 13. The concentrate of claim 10, wherein said vehicle is Subsequent introduction in said aqueous medium; and polyethylene glycol having a molecular weight within the d) mixing the non-aqueous solution and polysorbate 80 range from 8 to 1000. and octoxynol 40 mixture until a clear concentrate 14. The concentrate of claim 10, wherein said surfactant Solution is obtained. is polysorbate 80. 24. A concentrate solution for ophthalmic or perioph 15. The concentrate of claim 10, wherein said antifungal thalmic use containing from about 0.005% (w/w) to about agent is selected from the group consisting of amorolfine, 6.0% (w/w) antifungal agent, from about 24% (w/w) to amphotericin B, anidulafungin, butoconazole, butenafine, about 79% (w/w) non-aqueous vehicle, and from about 20% caspofungin, ciclopiroX olamine, clotrimazole, econazole, (w/w) to about 70.0% (w/w) surfactant. fluconazole, flucytosine, griseofulvin, haloprogin, itracona 25. A concentrate solution for ophthalmic or perioph Zole, ketoconazole, micafungin, miconazole, miconazole thalmic use containing from about 0.005% (w/w) to about nitrate, naftifine, nikkomycin Z, topical nystatin, liposomal 6.0% (w/w) antifungal agent tolnaftate or miconazole nystatin, oxiconazole, posaconazole, pimaricin, ravucona nitrate, from about 24% (w/w) to about 79% (w/w) non Zole, Sulconazole, terbinafine, terconazole, tioconazole, aqueous vehicle polyethylene glycol 400, and from about tolnaftate, undecylenate, and Voriconazole. 20% (w/w) to about 70.0% (w/w) surfactants polysorbate 80 16. An aqueous solution for ophthalmic or periophthalmic and octoxynol 40. use comprising an antifungal agent in an amount effective to 26. The concentrate solution of claim 24 or claim 25, inhibit fungal growth in said aqueous solution, said solution wherein said concentrate solution is dissolved in a first made by dissolving in first aqueous solution containing no aqueous solution to produce a second aqueous solution, antifungal agent a second concentrate solution containing (a) wherein said second aqueous solution is for ophthalmic or an antifungal agent dissolved in a physiologically acceptable periophthalmic use. non-aqueous vehicle, said vehicle being miscible with said 27. An aqueous solution for ophthalmic or periophthalmic first aqueous solution containing no antifungal agent; and (b) use comprising from about 0.001% (w/w) to about 2.0% one or more physiologically acceptable Surfactants; said (w/w) miconazole nitrate, from about 0.5 (w/w) to about concentrate having been added to the first aqueous solution 1.6% (w/w) polyethylene glycol 400, from about 0.3% containing no antifungal agent in an amount Sufficient to (w/w) to about 10.0% (w/w) polysorbate 80 and from about provide a fungicidally or fungistatically effective amount of 0.3% (w/w) to about 10.0% (w/w) octoxynol 40, and from said antifungal agent in said first aqueous Solution. about 0.0001% (w/w) to about 0.0002% (w/w) polyhexam 17. The method according to claims 1 or 7, wherein the ethylene biguanide. antifungal agent is tolnaftate. 28. An aqueous solution for ophthalmic or periophthalmic 18. The method according to claims 1 or 7, wherein the use comprising from about 0.001% (w/w) to about 2.0% antifungal agent is miconazole nitrate. (w/w) miconazole nitrate, from about 0.5% (w/w) to about 19. The method according to claims 1 or 7, wherein said 1.6% (w/w) polyethylene glycol 400, from about 0.3% vehicle or solvent is polyethylene glycol 400. (w/w) to about 10.0% (w/w) polysorbate 80 and from about 20. The method according to claims 1 or 7, wherein said 0.3% (w/w) to about 10.0% (w/w) octoxynol 40, and from surfactants are polysorbate 80 and octoxynol 40. about 0.001% (w/w) to about 0.002% (w/w) polyguater 21. The method according to claims 1 or 7, wherein said nium-1. aqueous solution is a multipurpose lens solution. 29. An aqueous solution for ophthalmic or periophthalmic 22. The aqueous solution of claim 16, wherein the anti use comprising from about 0.001% (w/w) to about 2.0% fungal agent is selected from the group consisting of amo (w/w) tolnftate, from about 0.5% (w/w) to about 1.6% (w/w) rolfine, amphotericin B, anidulafungin, butoconazole, buten polyethylene glycol 400, from about 0.3% (w/w) to about afine, caspofungin, ciclopiroX olamine, clotrimazole, 10.0% (w/w) polysorbate 80 and from about 0.3% (w/w) to econazole, fluconazole, flucytosine, griseofulvin, halopro about 10.0% (w/w) octoxynol 40. gin, itraconazole, ketoconazole, micafungin, miconazole, 30. A method for avoiding fungal infection in a subject, miconazole nitrate, naftifine, nikkomycin Z, topical nystatin, said infection arising from contact of the eye of said subject liposomal nyStatin, oxiconazole, posaconazole, pimaricin, with an aqueous Solution during ophthalmic or perioph raVuconazole, Sulconazole, terbinafine, terconazole, tio thalmic use of said aqueous solution, the method compris conazole, tolnaftate, undecylenate, and Voriconazole. ing: providing as the aqueous solution used ophthalmically 23. A concentrate solution containing 1.0% (w/w) or periophthalmically an aqueous Solution Suitable for oph miconazole nitrate in a form Suitable for Subsequent intro thalmic or periophthalmic use to which has been added a duction in an aqueous medium suitable for ophthalmic or concentrate comprising (a) an antifungal agent dissolved in periophthalmic use, said concentrate solution made by: a non-aqueous vehicle, said vehicle being miscible with an aqueous solution Suitable for ophthalmic or periophthalmic a) dissolving 1.0% (w/w) miconazole nitrate in 50% use containing no antifungal agent, and (b) one or more (w/w) polyethylene glycol with continual mixing at a physiologically acceptable surfactants, said concentrate hav temperature of 75° C.-85° C. to form a non-aqueous ing been added in an amount Sufficient to provide a fungi Solution of said antifungal agent upon its Subsequent cidally or fungistatically effective amount of said antifungal introduction in said aqueous medium, said vehicle being miscible with aqueous Solutions; agent in said provided aqueous solution. b) cooling said non-aqueous solution to 30° C.-40°C.; k k k k k