OA12808A - Use of topical compositions for the control of microbial diseases of the nail. - Google Patents

Abstract

A method of treating or preventing a microbial disease of a subject's nail, such as onychomycosis, the method comprising the step of topically administering to the nail an effective amount of copper silicate.

Description

012808 -1 -

Use of Topical Compositions for the Control of Microbial Diseases ofthe

Mail

Field of the Invention

The présent invention relates to the use of copper silicate for treating and 5 preventing microbial infections and diseases and in particuiar fungal diseases ofthe naii such as onychomycosis. The présent invention also relates to coppersilicate compositions adapted for topical administration.

Backqround Art

Microbial diseases of the naii are a considérable public health concern. Whilst 10 there are diseases and disorders caused by baçteria, the most prévalent andclinically significant naii diseases are caused by fungi.

Onychomycosis is a fungal disease of nails caused by a range of fungi, principallydermatophytes - Trichophyton species such as Trichophyton mentagrophytes andTrichophyton rubrum, but also Trichophyton megninii, Trichophyton schoenieinii 15 and Trichophyton tonsurans and Candida species, such as C. aibicans. Infectionis of the naii unit (the naii matrix, bed or plate) and although not life threatening itcan cause inconvenience, pain, discomfort and sometimes serious physical andoccupational limitations.

There is presently a range of oral médications avaiiable for' treating 20 - onychomycosis. However, these are generaliy expensive, suffer from limitedeffectiveness, require close mônitoring from practitioners and can cause seriousside effects such as iiver and heart damage.

Whilst there is a clear need for an effective topical agent for treatingonychomycosis, the agents presently avaiiable hâve iimited efficacy. The most 25 widely used topical agent is Penlac®. However, the clinical studies concerningthis agent report low efficacy. 012808 -2-

The main difficulty faced when deveioping an effective topical agent for the naii isthat the nail plate is thick, hard and dense, and represents a formidable barrier fordrugs preventing them from reaching the infection site in a therapeuticallyrequired quantity. Adding to this problem is the fact that onychomycosis is 5 associated with a thickening of the naii plate. To overcome this problem prior arttopical agents for treating onychomycosis hâve incorporated agents that increasethe permeability of the naii plate. However, these topical agents contain multipleagents i.e, a softener and an anti-fungal and thus are reiatively expensive toproduce. Furthermore, such agents hâve met with limited success. 10 The présent invention seeks to overcome the above problems by providing a safeand effective topical anti-microbial that is capable of treating microbial infectionsof the nail.

Summarv of the Invention

The présent invention provides a method of treating or preventing microbial 15 infection of a subject’s nail, the method comprising the step of topîcallyadministering to said nail an effective amount of copper silicate.

The infections treated or prevented by the method of the présent invention maymanifest themselves into disease States that may also be treated according to theprésent invention. Thus, the présent invention aiso provides a method of treating 20 or preventing a microbial disease of a subject’s nail, the method comprising thestep of topically administering to the nail an effective amount of copper siiicate.

The présent invention also provides a composition adapted for topicaladministration comprising an anti-microbial effective amount of copper siiicate.

Brief Description of the Figures 25 Figure 1 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6 and week 12) of a dermatophyte infection of a first subject’snaii, according to one exemplary method of the présent invention; 012808 -3-

Figure 2 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6 and week 12) of a dermatophyte infection of. a second subject'snail, âccording to one exempiary method of the présent invention;

Figure 3 is a sériés of photos showing the time course of treatment (Day 0:untreated and week 6) of a dermatophyte infection of a third subject’s nail,âccording to one exempiary method of the présent invention;

Figure 4 is a sériés of photos showing the time course of treatment (Day 0:untreated and week 6) of a dermatophyte infection of a fourth subject’s nail,âccording to one exempiary method of the présent invention;

Figure 5 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 12 and week 12 - post-treatment) of a dermatophyte infection ofa fifth subject’s nail, âccording to one exempiary method of the présent invention;

Figure 6 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6, week 26 and week 12 - post-treatment) of a dermatophyteinfection of a sixth subject’s nail, âccording to one exempiary method of theprésent invention; and

Figure 7 is a sériés of photos showing the extended time course of treatment(Day 0: untreated, week 6, week 26 and week 12 - post-treatment) of adermatophyte infection of the second subject's nail, âccording to one exempiarymethod of the présent invention.

Petaifed Description of the Invention

The présent invention provides a method of treating or preventing mîcrobialinfection of a subject’s nail, the method comprising the step of topicallyadministering to said nail an effective amount of copper silicate.

For the purposes of the présent invention the term “topically administering”includes, but is not fimited to, application through the nail plate with a device 012808 -4- adapted to perforate the nail plate and application between the nail plate and nailbed with a device adapted to be inserted therebetween.

For the purposes of the présent invention the term "nail” refers to one or more ofthe éléments of the nail unit (the nail matrix, bed or plate) and also includes toe 5 and fingemails and animal nail like features such as hooves, trotters and claws.

Surprisingly, it has been found that the copper silicate administered to a nailaccording to the présent invention delivers either an effective anti-microbial doseto the infected area or a dose sufficient to prevent Infection with a mlcrobial nailpathogen. In this regard, the applicant has discovered that upon administration of 10 the copper silicate to the nail, therapeutically effective amounts of the copper areable to reach the site of infection. The précisé mode of this effect has not beenestablished conclusively, but the Applicant is aware that the copper silicatepersists aî the site of application and/or the site of infection.

Topical administration may be achieved in any one of a number of ways. 15 Preferabiy, the compound is applied to the nail by painting, wiping, dabbing orspraying the nail with the compound. Aiternatively, topical administration may beachieved by submerging the nail in a copper silicate solution or contacting the nailwith a means for dispensing an effective amount of copper silicate. Dispensingmeans can be varîed and include slow release carriers and materials impregnated 20 with copper silicate such as patches, bandages, cotton wool and gauze.Preferabiy, the dispensing means is adapted to be removably fixed to the nail todeliver the copper silicate over a predetermined time and/or at a predetermineddose.

The methods of the présent invention may be used to treat mlcrobial, such as 25 fungal, infection in any animal and more particularly any mammal. However,preferabiy, the subject is human. Thus the présent invention also provides for theuse of copper silicate for preparing a médicament for treating microbial infectionssuch as fungal infections including onychomycosis. 012808 -1-

Use of Topical Compositions for the Control of Microbial Diseases of the

Nail

Field of the Invention

The présent invention relates to the use of copper silicate for treating and 5 preventing microbial infections and diseases and in particular fungal diseases ofthe nail such as onychomycosis. The présent invention also relates to coppersilicate compositions adapted for topical administration.

Background Art

Microbial diseases of the nail are a considérable public health concern. Whilst 10 there are diseases and disorders caused by bacteria, the most prévalent andclinically significant nail diseases are caused by fungi.

Onychomycosis is a fungal disease of nails caused by a range of fungi, principallydermatophytes - Trichophyton species such as Trichophyton mentagrophytes andTrichophyton rubrum, but also Trichophyton megninii, Trichophyton schoenleinii 15 and Trichophyton tonsurans and Candida species, such as C. aibicans. Infectionis of the nail unit (the nai! matrix, bed or plate) and although not life threatening itcan cause inconvenience, pain, discomfort and sometimes serious physical andoccupational limitations. —There is presently a range of oral médications available for treating 20 onychomycosis. However, these are generally expensive, suffer from limitedeffectiveness, require close monitoring from practitioners and can cause seriousside effects such as liver and heart damage.

Whilst there is a clear need for an effective topical agent for treatingonychomycosis, the agents presently available hâve limited efficacy. The most 25 widely used topical agent is Penlac®. However, the clinical studies concerningthis agent report low efficacy. 012808 - 2 -

The main difTiculty faced when developing an effective topical agent for the nail isthat the nail plate is thick, hard and dense, and represents a formidable barrier fordrugs preventing them from reaching the infection site in a therapeuticallyrequired quantity. Adding to this problem is the fact that onychomycosis is 5 associated with a thickening of the nail plate. To overcome this problem prior arttopical agents for treating onychomycosis hâve incorporated agents that increasethe permeability of the nail plate. However, these topical agents contain multipleagents i.e. a softener and an anti-fungal and thus are relatively expensive toproduce. Furthermore, such agents hâve met with limited success. 10 The présent invention seeks to overcome the above problème by providing a safeand effective topical anti-microbial that is capable of treating microbial infectionsof the nail.

Summary of the Invention

The présent invention provides a method of treating or preventing microbial 15 infection of a subject’s nail, the method comprising the step of topicallyadministering to said nail an effective amount of copper silicate.

The infections treated or prevented bÿ the method of the présent invention maymanifest themselves into disease states that may also be treated according to theprésent invention. Thus, the présent invention also provides a method of treating 20 or preventing a microbial disease of a subject’s nail, the method comprising thestep of topically administering to the nail an effective amount of copper silicate.

The présent invention also provides a composition adapted for topicaladministration comprising an anti-microbial effective amount of copper silicate.

Brief Description of the Figures 25 Figure 1 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6 and week 12) of a dermatophyte infection of a first subject’snail, according to one exemplary method of the présent invention; 012808 -3-

Figure 2 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6 and week 12) of a dermatophyte infection of a second subject’snail, according to one exemplary method of the présent invention;

Figure 3 is a sériés of photos showing the time course of treatment (Day 0:5 untreated and week 6) of a dermatophyte infection of a third subject’s nail, according to one exemplary method of the présent invention;

Figure 4 is a sériés of photos showing the time course of treatment (Day 0:untreated and week 6) of a dermatophyte infection of a fourth subject’s nail,according to one exemplary method of the présent invention; 10 Figure 5 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 12 and week 12 - post-treatment) of a dermatophyte infection ofa fifth subject’s nail, according to one exemplary method of the présent invention;

Figure 6 is a sériés of photos showing the time course of treatment (Day 0:untreated, week 6, week 26 and week 12 - post-treatment) of a dermatophyte 15 infection of a sixth subject’s nail, according to one exemplary method of theprésent invention; and

Figure 7 is a sériés of photos showing the extended time course of treatment(Day 0: untreated, week 6, week 26 and week 12 - post-treatment) of adermatophyte infection of the second subject’s nail, according to one exemplary 20 method of the présent invention.

Detailed Description of the Invention

The présent invention provides a method of treating or preventing microbia!infection of a subject’s nail, the method comprising the step of topicailyadministering to said nail an effective amount of copper silicate. 25 For the purposes of the présent invention the term “topicaily administering"includes, but is not limited to, application through the nail plate with a device 012808 -4- adapted to perforate the nail plate and application between the nail plate and nailbed with a device adapted to be inserted therebetween.

For the purposes of the présent invention the term “nail" refers to one or more ofthe éléments of the nail unit (the nail matrix, bed or plate) and also includes toe 5 and fingernails and animal nail like features such as hooves, trotters and claws.

Surprisingly, it has been found that the copper silicate administered to a nailaccording to the présent invention delivers either an effective anti-microbial doseto the infected area or a dose sufficient to prevent infection with a microbial nailpathogen. In this regard, the applicant has discovered that upon administration of 10 the copper silicate to the nail, therapeutically effective amounts of the copper areable to reach the site of infection. The précisé mode of this effect has not beenestablished conclusively, but the Applicant is aware that the copper silicatepersiste at the site of application and/or the site of infection. 'Topical administration may be achieved in any one of a number of ways. 15 Preferably, the compound is applied to the nail by painting, wiping, dabbing orspraying the nail with the compound. Alternatively, topical administration may beachieved by submerging the nail in a copper silicate solution or contacting the nailwith a means for dispensing an effective amount of copper silicate. Dispensingmeans can be varied and include slow release carriers and materials impregnated 20 with copper silicate such as patches, bandages, cotton wool and gauze.Preferably, the dispensing means is adapted to be removably fixed to the nail todeliver the copper silicate over a predetermined time and/or at a predetermineddose.

The methods of the présent invention may be used to treat microbial, such as 25 fungal, infection in any animal and more particularly any mammal. However,preferably, the subject is human. Thus the présent invention also provides for theuse of copper silicate for preparing a médicament for treating microbial infectionssuch as fungal infections including onychomycosis. -5- 012808

It is common for nails to be infected with a range of microbes such as bacteriaand fungi (including yeast). Thus, the microbial infection may be bacterial.However, the method of the présent invention is particuiarly usefu! for treatingfungal infections. 5 The amount of copper silicate applied in the method of the présent invention willbe sufficient to effectiveiy treat or prevent the infection and thus will necessarilyvary depending at least on the severity and type of the infection, the strength ofthe composition and the manner in which individual patients respond to thetreatment However, preferably a composition containing 1-100 pg of copper is 10 applied to each infected nail, more preferably a composition containing 2-50 pg ofcopper is applied to each infected nail and even more preferably, a compositioncontaining 5-20 pg of copper is applied to each infected nail. In one particularform of the invention a composition containing at least about 10 pg of copper isapplied to each infected nail. 15 'Similarly, the frequency with which, and the duration for which, the copper silicateis applied will be sufficient to effectiveiy treat or prevent the infection and thus willalso vary depending at least on the severity and type of infection, the strength ofthe composition and the manner in which individual patients respond to thetreatment. Preferably, the copper silicate is applied at least once, twice or three 20 times a day for at least 1,3, 6,12, 26, 30, 40, 45, 48 or 52 weeks. Alternatively, itmay be applied once every 2 - 14 days for as long as necessary to treat orprevent infection.

Thus, the présent invention also provides a method of treating or preventingmicrobial infection of a subject’s nail, the method comprising the step of 25 repeatedly topically administering to said nail an effective amount of coppersilicate.

As indicated above the method of the présent invention may be applied to treatfungal infections. In this regard, fungal infections which may be treated accordingto the présent invention include infections from dermatophytes, such as 30 Trichophyton species including Trichophyton mentagrophytes fincluding varieties 012808 -6- mentagrophytes and interdigitales), Trichophyton rubrum, Trichopbyton megninii,Tnchophyton schoenleinii, Trichophyton tonsurans; yeasts, such as Candidaspecies, most notably Candida albicans, Candida glabrata, Candida ciferrii;Microsporum species including Microsporum canis and Microsporum gypseum;Malassezia species including Malassezia furfur and Malassezia sympodialis;Epidermophyton species such as Epidermophyton floccosum; Scopularosisspecies; Acremonium species and Aspergillus species.

Thus, the présent invention also provides a method of treating or preventinginfection of a subject’s nail with one or more of: dermatophytes, such asTrichophyton species including Trichophyton mentagrophytes (including varietiesmentagrophytes and interdigitales), Trichophyton rubrum, Tnchophyton megninii,Trichophyton schoenleinii, Trichophyton tonsurans; yeasts, such as Candidaspecies, most notably Candida albicans, Candida glabrata, Candida ciferrii;Microsporum species including Microsporum canis and Microsporum gypseum;Malassezia species including Malassezia furfur and Malassezia sympodialis;Epidermophyton species such as Epidermophyton floccosum; Scopularosisspecies; Acremonium species and Aspergillus species, the method comprisingthe step of topically administering to said nail an effective amount of coppersilicate.

Fungal nail infections that may be treated using the methods of the présentinvention are usually characterized by tarnished white, yellowed, or blackenednails. The nails may pull away from the pink nail bed along the sides or outer - edges, and infections are usually exacerbated by hot, damp conditions, such asinside footwear or in environments where hands or feet are continually exposed tomoisture. The fungal infections can spread from toe to toe, foot to foot, and footto hand.

Methods of treating or preventing microbial diseases

The présent invention also provides a method of treating or preventing a microbialdisease of a subject’s nail, the method comprising the step of topicallyadministering to the nail an effective^mount of copper silicate. 012808 -7-

Whilst the method of îhe présent invention may be applied to treat a wide rangeof microbial diseases of the nail it is particulariy useful for treating fungal diseasesof the nail. Thus, the présent invention also provides a method of treating orpreventing a fungal disease of a subject's nail, the method comprising the step of 5 topically administering to the nail an effective amount of copper silicate.

Spécifie fungal diseases that can be treated by the methods of the présentinvention include onychomycosis such as distal subungual onychomycosis,superficial white onychomycosis, proximal white subungual onychomycosis; totalsecondary dystrophie onychomycosis; and total dystrophie primary 10 onychomycosis.

In some situations, it may be désirable to combine the method of treating orpreventing a fungal disease according to the présent invention with otherthérapies. Thus, the présent invention also provides a method of treating orpreventing a fungal disease of a subject’s nail, the method comprising the step(s) 15 of topically administering to said nail an effective amount of copper silicate and (ii)administering to said subject an effective amount of at least one other anti-fungalagent.

The other anti-fungal agent may be varied and includes, for example, one or moreorally administrable anti-fungal agents selected from the group consisting of: 20 miconazole, kétoconazole, itraconazole, fluconazole, econazole, ciclopirox,oxiconazole, clotrimazole, terbinafine, naftifine, pharmaceutically acceptable salts _ . thereof and stereoisomers thereof, Lamisil™, Sporanox™ and Diflucan™.

The présent invention also provides for the use of copper silicate for preparing amédicament for treating or preventing a disease caused by a fungal infection. 25 Topical Formulations

The présent invention also provides a composition adapted for topicaladministration to a nail comprising an anti-microbial effective amount of coppersilicate. 012808 -8-

The form of the composition of the présent invention may also be varied providedit retains its anti-microbial properties. Preferably, the composition is a solution.However, the composition may also be in solid form provided it is properlyformulated. In this regard, the composition couid comprise copper silicate in the 5 form of a micronized solid such as chrysocolla.

When the copper silicate is provided in the form of a solution, it preferably isprovided as an aqueous acidified solution. Acidified solutions are particularlypreferred because copper silicate is more soluble at acidic pH. Particularlypreferred pHs are 3-6, 4-6 and 5-6. In one example, the copper silicate is 10 prepared according to the methods described and claimed in US patent5,474,972.

The composition adapted for topical administration may be in the form of any oneof the following: solution, lotion, suspension, émulsion, cream, gel, ointment,liniment and salve. Particularly preferred forms are ointments, creams or gels. 15 Ointments generally are prepared using either (1) an oleaginous base, i.e., oneconsisting of fixed oils or hydrocarbons, such as white petroleum or minerai oil, or(2) an absorbent base, i.e., one consisting of an anhydrous substance orsubstances that can absorb water, for example anhydrous lanolin. Customarily,following formation of the base, whether oleaginous or absorbent, the active 20 ingrédient is added to an amount affording the desired concentration.

Creams are oil/water émulsions. They consist of an oil phase (internai phase),comprising typically fixed oils, hydrocarbons and the like, waxes, petroleum,minerai oil and the like and an aqueous phase (continuous phase), comprisingwater and any water-soluble substances, such as added salts. The two phases 25 are stabilised by use of an emulsifying agent, for example, a surface active agent,such as sodium lauryl sulfate; hydrophilic colloids, such as acacia colloïdal clays,veegum and the like. For the purposes of the présent invention, the compoundmay be added to the water phase prior to formation of the émulsion, in an amountto achieve the desired concentration. 012808 -9-

Gels comprise a base selected from an oieaginous base, water, or an emulsion-suspension base. To the base is added a gelling agent that forms a matrix in thebase, încreasing its viscosity. Examples of gelling agents are hydroxypropylcellulose, acrylic acid polymers and the like. For the purposes of the présent 5 invention the compound may be added to the formulation at the desiredconcentration at a point preceding addition of the gelling agent.

The compositions of the présent invention may be produced by dissolving orcombining the copper silicate in an aqueous or non-aqueous carrier. In general,any liquid, cream, or gel, or similar substance that does not appreciably react with 10 the silicate or any other active ingrédient that may be introduced and which isnon-irritating is suitable.

Thus, the présent invention also provides a method of producing copper silicateadapted for topical administration comprising the step of dissolving or combiningthe copper silicate in an aqueous or non-aqueous topical carrier. 15 The composition of the présent invention may further comprise an auxiliary agentsuch as any one or more of: preservatives, stabilizers, emulsifiers, wetting agents,fragrances, colouring agents, odour controllers and thickeners such as naturalgums.

The concentration of the copper in the composition may be varied depending at 20 least on the severity and type of infection that the composition is to be used totreat or prevent. However, preferably, the concentration of the copper isapproximately 100 mg/L - 10 g/L (as Cu). More preferably, the copperconcentration is to a final concentration of approximately 1 g/L - 5 g/L or 2g/L - 3g/L (as Cu). 25 The présent invention will now be described with reference to the followingexamples. The description of the examples is in no way to limit the generality ofthe preceding description. 012808 -10-

Examples

Example 1 : In vitro funigicidal activitv of a copper silicate solution

Trials were carried out to assess the fungicidal activity of copper silicate against arange of fungi. The results of the study are set out in Tables 1a and 1b 5 hereunder.

Table 1a - Minimum inhibitory (MIC) and fungicidal (MFC) concentrations of acopper silicate solution (ppm Cu) for Candida spp., Malassezia spp. and variousdermatophytes.

Orgaiiism N MIC* MFC* Candida albicans 8 128 1024 Candida ciferrii 2 128 128 Candida glabrata 2 256 256 Malassezia furfur 6 512 >1024 Malassezia sympodialis 7 512 >1024 Epidermophyton floccosum 10 256 256 Microsporum canis 10 128 256 Microsporum gypseum 10 128 256 Trichophytonmentagrophytes varinterdigitales 10 256 256 Trichophytonmentagrophytes varmentagrophytes 10 256 256 Trichophyton rubrum 10 128 256 Trichophyton tonsurans 10 128 256 *Modal values 10 Table 1b: Minimum Inhibitory Concentration (MIC) and Minimum FungicidalConcentration (MFC) of copper silicate (% w/w as Cu) for various dermatophytesand non-dermatophytes that cause onychomycosis, and Candida spp.

Species

Trichophyton rubrumT. mentagrophytesvar interdigitaleT. mentagrophytesT. tonsurans

Epidermophyton floccosum 26 19 12 10 11 MIC* MFC* 0.0175 0.0350 0.0175 0.0350 0.0175 0.0350 0.0350 0.0350 0.0350 0.0350 012808 -11 -

Microsporum canis 9 0.0350 0.0700 M. gypseum 9 0.0350 0.1400 Aspergillus flavus 12 0.1400 >0.1400 A. fumigatus 12 0.0350 >0.1400 A. niger 14 0.0350 0.1400 A. terrus 7 0.0700 >0.1400 Fusarium spp. 16 0.0700 0.0700 Scopulariopsis spp. 5 0.0350 >0.1400 Scedosporium spp. 5 0.0350 >0.1400 Candida afbicans 10 0.0350 0.0700 C. tropicalis 10 0.0350 0.0700 C. parasi/osis 10 0.0350 0.0350 C. g/abrate 4 0.0350 0.1400 C. pseudotropicalis (kefyr) 1 0.0350 0.0350 C. krusei 1 0.0175 0.0175

Modal Values

Example 2: In vitro bactericidal activity of a copper silicate solution

Table 2: Minimum inhibitory concentrations (MIC) of copper silicate solution (ppmCu) for a variety of bacteria

Organisms n Range M1Cm MetR - Staphylococcus aureus 13 256-512 512 Mets - Staphylococcus aureus 9 256-512 NA Propionibacterium acné 19 4-256 256 Staphylococcus saprophyticus 4 1024 NA Streptococcus pneumoniae 27 ‘ 128-512 512 Streptococcus pyogenes 11 256 - 1024 512 Streptococcus spp. 8 128 - 1024 NA Acinetobacter spp. 13 512-1024 1024 Enterobacteriaceae * 113 1024- >1024 >1024 Enterococcus faecalis 12 64 - 256 256 Enterococcus faecium ' 2 128-256 NA Morganella morganii 7 >1024 NA ~Moraxella catarrhalis 3 <6 NA Pseudomonas aeruginosa 13 1024 ->1024 >1024 Stenotrophomonas maltophilia 12 1024-> 1024 >1024

Enterobacteriaceae spp. include Citrobacter freundîi (n=11), Enterobacter aerogenes (n=13),Enterobacter cloacae (n=13), Escherichia coli (n=25), Klebsiella oxytoca (n=10), Klebsiellapneumoniae (n=24), Profeus mirabilis (n=12) and Providencia spp.(n=5)

Example 3: Treatment of onychomycosis with copper silicate

This study évaluâtes copper silicate as a topical fungicide as a treatment ofonychomycosis and compares a solution and cream formulation. -12- 012808

Materials/Methods (A) Fungicidal solution (CSSOL1)

Ingrédients

Sodium silicate solution (30% SiO2,10% Na2O) 5 Empimin LSM 30AU™ - Huntsman International LLC (sodium alkyl ether sulfate)

Copper sulfate pentahydrateAcetic Acid (90% solution)

Water (deionised)

Technical Instructions 10 1. The solution is prepared as two separate parts (A and B) as follows: - Part A, Silicate Solution

Product Welght (g)

Water 56.7

Sodium silicate solution 60.8 (30% SiO2) 10% Na2O)

Empimin LSM 30AU™ 2.5 /ora/ 120g (~100ml)

Method: Add the sodium silicate solution (30% SiO2, 10% Na2O) and empiminLSM 30AU ™ to the water with stirring. Filter.

Part B. Copper Solution

Product Weight (g)

Water 88.2

Copper sulfate pentahydrate 11.5

Acetic acid (90%) 9.4

Total 109.1g (-100ml) 012808 -13-

Method: Add the copper sulfate pentahydrate to the water with stirring andcontinue stirring until a clear solution is obtained. Add the acetic acid with stirring.Filter. 2. The final solution is then produced by dilution of the combined parts. 5

Product Weight (g)

Water 880.0

Part A 24.0

Part B 109.1

Total 1013.1 (~1000mL)

Method: Add Part A to the water with stirring. Continue to stir while adding PartB. Filter.

The final product should be a light blue, clear solution with an acrid odour.

Product Spécifications: 10 pH: 3.3-3.7

Copper content: 2.8 +/- 0.28 g/L as Cu (B) Copper silicate solution concentrate for use in creams (CSC1)

Ingrédients Required

Sodium silicate solution (30% SiO2,10% Na2Û) 15 Copper sulfate pentahydrate

Acetic Acid (90% sofution)

Water (deionised)

Technical Instructions: 1. The solution is prepared as two separate parts (A and B) as follows: 20 Part A. Silicate Solution -14- 012808

Product

Water

Sodium silicate solution(30% SiO2, 10% Na2O)

Weight (g)59.260.8

Total 120g (~100mL)

Method: Add the sodium silicate solution (30% SiO2,10% Na2O) to the water withstirring. Filter.

Part B. Copper Solution

Product Weight (g)

Water 88.2

Copper sulfate pentahydrate 11.5

Acetic acid (90%) 9.4

Total 109.1g (~~100mL) 5 Method: Add the copper sulfate pentahydrate to the water with stirring andcontinue stirring untii a clear solution is obtained. Add the acetic acid with stirring.Filter. 10 2. The final concentrated solution is then produced by dilution of thecombined parts. CSC1 Concentrate Solution

Product

WaterPart APart B

Weight (g)130.024.0109.1

Total 263.1 (~250mL)

Method: Add Part A to the water with stirring. Continue to stir while adding PartB. Filter.

Copper content: 11.2 g/L as Cu 15 The final product should be a iight blue, clear solution with an acrid odour. 012808 -15- (C) Funqicidal cream 1. Formulation ingrédient %wt Phase A. Water 42.77 Copper silicate conc. 20.00 (CSC1 - pH adjusted to 5.0 with triethanolamine) Propylene glycol 5.00 Germaben IIE™ (Sutton Laboratories 0.50 Inc) Brij 78™ (Atlas Powder Company) 3.00 Sorbic Acid 0.10 Red no. 33 Solution (0.02%) 0.20 Phase B. Propyl paraben 0.20 Tween 60™ (Atlas Powder Company) 2.56 Stearyl alcohol 3.61 Ceteth-20 3.00 Cetyl alcohol 2.56 Cetearyl alcohol 2.00 Myristyl myristate 2.00 PEG-4 Olivaie 5.00 Octyl palmitate 4.00 Glyceryi stéarate (and) PEG-100 stéarate 3.00 20% Titanium dioxide paste 0.50 Total 100.00 5 2. Method a. Into the main mixing vessel which shouid be fitted with a variable speed“Silverson” mixer and a sweep-blade contra-rotating agitator, weigh theingrédients of Phase B. Stir until uniform. Adjust température to 75°C. 10 b. into a separate vessel weigh the phase A ingrédients, mix with a propellerstirrer until ail solids are dissolved. Adjust température to 75°C. c. Add phase A to phase B, using the “Silverson" mixer to form a smootheven émulsion. d. Switch on the sweep-blade agitator and begin cooling the batch. 15 e. Continue cooling and mixing until the température is 35°C. f. pH shouid be 5.3 - 5.5.

Subjects, not already receiving anti-fungal treatment, were assessed for clinicaldiagnosis of onychomycosis and photographe and nail clippings were taken.Those testing positive for onychomycosis were admitted to the trial. -16- 012808

The anti-fungal composition (cream or solution) was applied to the infected nailtwice daily for the duration of the treatment.

Estimâtes of the nail area involved were made during clinical visits andphotographs were taken. Nail clippings were collected for KOH and 5 dermatophyte culture. Subjects were assessed by a clinician for onychomycosisusing the following criteria: • “Not effective” is >10% nail involvement with KOH positive and culturepositive; • “Mycological cure” KOH négative and culture négative; 10 · “Effectively treated” is <10% nail involvement, with KOH and culture négative;and • “Complété cure” is clinical clearance of the target nail together with négative--- KOH and culture.

Results 15 The results presented hereunder are intérim results of a 12 month study.Patients entered the trial and commenced their treatment at different dates. Ofthe 18 patients enrolled in the trial, 13 had reached 6 weeks of treatment, and ofthese, 10 had reached 12 weeks of treatment, at the time of reporting theseresults. Patients were assessed at 0 (commencement), 6 weeks and 12 weeks. 20 The results of these assessments are tabulated below in Table 3. No différenceswere noted in efficacy between the cream and solution at any of theassessments. 012808 -17-

Table 3

Criteria % of Patients Day 0 (n=18) 6 Weeks (n=13) 12 Weeks (n=10) Mycology (KOH négativeor culture négative) 0 53 80 Mycology (KOH négativeand culture négative) 0 0 20 Clinical - New growthclear 0 61 70 Clinical -100% clear 0 0 30 Complété cure (KOH négative, culturenégative and 100%clear) 0 0 20

The effectiveness of the treatment method in the exampie is exempiified inFigures 1-4 that contain photographe of patient’s nails treated in the triai.

No adverse reactions were reported 5 Exampie 4: Treatment of onychomycosis with copper silicate

The triai reported in Example 3 was çontinued and the updated results are set outhereunder in Tables 4, 5 and 6 which list the clinical, mycoiogical and efficacymeasures, respectively.

Table 4

Treatment tinte (N=18) Post-treatment (N-18) Day 0 Week 6 Week 12 Week 26 Week 13 Week 26 % Nail Infected 0% (clear) 0 0 3 3 3 3 1-10% 1 2 1 1 1 2 11-50% 5 5 4 8 6 6 >50% 12 11 10 6 8 7 012808 15 15 3 3 -18-

New growth

Clear 10 11

Not clear 8 7 16 2 16 2

Note: One of the “not clear" patients had a damaged nail plate and was always assessed as “notclear".

Table 5

Treatment tîme (N=18) Post-treatment (N=18) Day 0 Week 6 Week 12 Week 26 Week 13 Week 26 KOH Positive 17 14 14 12 14 12 Négative 1 4 4 6 4 6 Dermatophvte culture Positive 18 8 7 6 8 9 Négative 0 10 11 12 10 9 Infecting orqanism T. rubrum 12 7 6 5 7 8 T. mentagrophytes v.interdigftale 6 1 1 1 1 1

Note: Between 39 weeks and 52 weeks (26 weeks post treatment) another patient who had beenassessed as culture négative reverted to culture positive with growth of T. rubrum beingrecorded. Ail 3 “clinical cure" patients were infected with T. mentagrophytes v. interdigitale.

Table 6

Treatment tinte Post-treatment Outcomes Week 6 Week 12 Week 26 Week 13 Week 26 . (N=18) (N=18) (N=18) (N=18) (N=18) Complété cure 0 2 3 3 3 Effective treatment 2 3 3 3 4 Mycological cure 4 3 5 3 4 Mycological change 6 8 7 7 5 Not effective 8# 6@ 8@ 9@ includes 2 patients that are KOH négative 'includes 1 patient that is KOH négative -19-

The effectiveness of the treatment method in the example is exemplified inFigures 5-7 that contain photographs of patient’s nails treated in the trial.

No adverse reactions hâve been reported.

Summary

At week 26, 3 patients achieved the primary endpoint of “complété cure" andmaintained their status 26 weeks post-treatment at 52 weeks. By 26 weeks posttreatment 4 (22%) patients were “effectively treated", an increase by one patientover 26 and 39 weeks which only included the 3 “complété cure” patients. “Mycological cure” was achieved in 5 (28%) patients after 26 weeks, but 2 ofthese had reverted to KOH positive at 39 weeks. At 52 weeks the “mycologicalcure” rate was 22%. “Mycological change”, or the change from culture positive to culture négative, wasachieved by another 7 patients at weeks 26 and 39. This gave a total of 12culture négative patients at 26 weeks and 10 at 39 weeks.

In total, 67% of patients were culture négative at the end of treatment, 55% at 13weeks post-treatment, and 50% (9 patients) ai the end of the trial.

The primary and secondary endpoints of “complété cure” and “effectivetreatment” are both dépendent upon the mycological cure rate. “Mycologicalcure" is defined as both KOH and culture négative, but patients may not becomeKOH négative until the diseased nail has grown out. It is usual for a great toenailto take 12-18 months to totally regrow. At the commencement of this trial, 12 ofthe 18 patients had >50% of their target nail infected, and of these 7 had cuticle(lunula) involvement. As a resuit of extensive disease, “mycological cure” andthus “effective treatment" and “complété cure” may not hâve been achievable bythese patients in the time allotted to this study. 012808 -20-

The treatment time of 26 weeks is considerably shorter than for most topicals thatare applied for a minimum of 48 weeks. It is évident from this trial that patientswith >50% of the nail plate diseased would hâve benefited from an extendedtreatment time. The results at 26 weeks show that many (50%) had significant 5 ciinical improvement with a réduction in the extent of their disease to <50% of thenail plate.

The présent invention includes any and ali modifications and adaptationsapparent to one skiiled in the art.

Throughout the spécification, unless the context requires otherwise, the word10 “comprise” or variations such as “comprises” or “comprising”, wilf be understoodto impiy the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

Claims (19)

1. 21 - 012808 The Claims Defining the Invention are as Foilows

   1. Use of copper silicate to préparé a médicament for treating a microbial naïlinfection.
2. 2. Use according to claim 1 wherein the microbial infection is a fungal naiiinfection.
3. 3. Use according to claim 1 or 2 wherein the infection comprises an infectionwith at least one dermatophyte.
4. 4. Use according to claim 3 wherein the dermatophyte is sefected from thegroup comprising Trichophyton spedes including Trichophytonmentagrophytes (including varieties mentagrophytes and interdigitales),Trichophyton rubrum, Trichophyton megninii, Trichophyton schoenleinii andTrichophyton tonsurans.
5. 5. Use according to claim 1 or 2 wherein the infection comprises an infectionwith at least a microbe sefected from the group comprising; Gandida species,Candida albicans, Candida glabrata, Candida cifeniï, Microsporum speciesincluding Microsporum canis and Microsporum gypseum-, Malassezia speciesincluding Malassezia furfur and Malassezia sympodialis; Epidermophytonspecies such as Epidermophyton floccosum; Scopufarosis species;Acremonium species and Aspergilius species.
6. 6. Use according to any one of the preceding claims wherein the médicamentcontains at least 1-100 pg of copper.
7. 7. Use according to any one of the preceding claims wherein the médicamentcontains at least 2-50 pg of copper.
8. 8. Use according to any one of the preceding claims wherein the médicamentcontains at least 10 pg of copper.
9. 9. Use according to any one of the preceding claims wherein the médicament isadapted to be applied at least 1-3 times a day. 22 012808
10. 10. Use according to any one of the preceding daims wherein the médicament isadapted to be applied at least once every 2-14 days.
11. 11. Use according to any one of the preceding daims where in the médicament isa topical médicament.
12. 12. Use of copper silicate to préparé a médicament for treating or preventing a disease selected from the group comprising: onychomycosis, distalsubungual onychomycosis, superficial white onychomycosis, proximal whitesubungual onychomycosis; total secondary dystrophie onychomycosis; andtotal dystrophie primary onychomycosis.
13. 13. Use of copper silicate for preparing a médicament for treating or preventing onychomycosis.
14. 14. Use according to daim 12 or 13 where in the médicament is a topicalmédicament.
15. 15. A composition adapted for topical administration to a nail comprising an anti- 15 microbial effective amount of copper silicate.
16. 16. A composition according to daim 15 in the form of a solution.
17. 17. A composition according to daim 16 in the form of an aqueous acidifiedsolution.
18. 18. A composition according to daim 15 in the form of a cream. 20 19. A method of producing a copper silicate composition adapted for topical - -administration comprising the step of dissolving or combining the copper silicate in an aqueous or non-aqueous topical carrier.
19. 20. A composition according to any one of daims 15 to 19 comprisingapproximately 100 mg/L -10 g/L (as Cu).