Topical compositions for the treatment of alopecia areata

Abstract

The disclosure relates to a pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally a co-solvent, for use in the topical treatment of alopecia areata.

Description

[0001] DRM23P03PC2

[0002] TITLE: TOPICAL COMPOSITIONS FOR THE TREATMENT OF ALOPECIA AREATA

[0003] Description

[0004] BACKGROUND OF THE INVENTION

[0005] Alopecia areata is an autoimmune disease affecting hair follicles, that is characterized by hair loss. Current treatments that are available for alopecia areata are directed at mitigating or containing the immunological attack processes associated with this disease, and / or towards the stimulation of hair regrowth. Available treatment options include the use of corticosteroids, minoxidil, phototherapy, platelet-rich plasma or contact allergens. However, such treatment options are associated also with mild, to in some cases, serious side effects.

[0006] Orally administered cyclosporine has been investigated as a treatment option for alopecia areata, either as a monotherapy or in combination with other treatment such as oral corticosteroids, however with variable outcomes. Oral, or in other words systemic treatments may however also result in unwanted or adverse side effects, in particular over long-term period of use, which may be necessary for treatment of alopecia areata and associated conditions, where therapeutic support may be required for the maintenance of hair regrowth. With only a first and also oral treatment for alopecia areata (baricitinib, a JAK inhibitor) only approved in 2022, there remains a need for an effective topical treatment of alopecia areata, without side effect profiles that is typically associated with systemic medication.

[0007] WO 2012 / 160179 A2 describes pharmaceutical compositions for topical administration comprising a semifluorinated alkane and an active ingredient for the treatment of a disease or condition affecting a skin or skin appendage. It is described that where the disease or condition affecting the skin is psoriasis, the treatment may be with an immunosuppressant drug as the active ingredient, with tacrolimus being a preferred compound.

[0008] It is thus an object of the present invention to provide a liquid pharmaceutical composition comprising cyclosporine which may be topically administered for use in the treatment of alopecia areata, for treating hair loss and / or for stimulating hair growth, and which provides for an effective, and moreover targeted treatment of said disease and associated conditions. It is also an object to provide a composition which may overcome one or more disadvantages of known compositions and treatments. Further objects of the invention will be clear on the basis of the following description of the invention, examples and claims. SUMMARY OF THE INVENTION

[0009] In a first aspect, the invention relates to a liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally a co-solvent, for use in the topical treatment of alopecia areata. In a further aspect, the invention relates to the pharmaceutical composition, for use in the topical treatment of hair loss. In yet a further aspect, the invention relates to the pharmaceutical composition for use in the topical stimulation of hair growth.

[0010] In yet another aspect, the invention relates to kit comprising a composition, or composition for use as described in any preceding claim, a container adapted for holding the composition and optionally a means for dispensing the composition, and / or instructions for use.

[0011] DETAILED DESCRIPTION OF THE INVENTION

[0012] In a first aspect, the present disclosure relates to a liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally, a co-solvent, wherein the composition is for use in the topical treatment of alopecia areata.

[0013] Alopecia areata is an autoimmune disease affecting hair follicles, that is characterized by hair loss. It is hypothesized that alopecia areata arises from a breakdown of immune privilege with respect to the hair follicle, with hair loss arising from a resulting state of active inflammation and immune attack on the hair follicle. This can result for example in weakening of the hair shaft, and disruption in the general normal homeostasis state of hair growth cycles. Biopsies of skin affected by alopecia areata typically show lymphocytic infiltrate around the bulb or lower part of the hair follicle.

[0014] Alopecia areata may be distinguished from other types of alopecia, such as androgenic alopecia, male pattern alopecia, female pattern alopecia, or other non-scarring types of alopecia such as, but not limited to trichotillomania, or mechanical traction- related alopecia. Alopecia areata may be transient In alopecia areata, the hair follicle is preserved and is nonscarring, scarring types of alopecia where the hair follicle is damaged and / or destroyed and where hair loss may be irreversible.

[0015] The liquid pharmaceutical compositions for use according to the present disclosure comprise, as a pharmaceutically active ingredient, cyclosporine. Cyclosporine (synonyms include cyclosporin A, CsA, or ciclosporin A) is an immunosuppressant drug that is widely used in connection with organ transplantation to reduce the activity of the patient’s immune system and thereby, the risk of organ rejection. Cyclosporine is thought to bind to the cytosolic protein cyclophilin (immunophilin) of immunocompetent lymphocytes, especially T-lymphocytes. This complex of cyclosporin and cyclophilin inhibits calcineurin, which, under normal circumstances, is responsible for activating the transcription of interleukin 2. It also inhibits lymphokine production and interleukin release and, therefore, leads to a reduced function of effector T-cells.

[0016] Cyclosporine is typically provided as a colourless or white powder. According to the present disclosure, the liquid pharmaceutical compositions comprise of cyclosporine dissolved in a semifluorinated alkane, or if a co-solvent is used, in a liquid pharmaceutical composition comprising cyclosporine dissolved in a mixture of a semifluorinated alkane and a co-solvent

[0017] In some embodiments, the concentration of cyclosporine in the composition according to the present disclosure is at a concentration of 1 to 30 mg / ml, 1 to 25 mg / ml, 1 to 10 mg / ml, 1 to 5 mg / ml, 5 to 20 mg / ml, or 3 to 15 mg / ml. Preferably, the concentration of cyclosporine in the composition according to the present disclosure is at a concentration of 8 to 12 mg / ml, or 9 to 11 mg / ml, more preferably the concentration of cyclosporine in the composition is about 10 mg / ml.

[0018] Semifluorinated alkanes are linear or branched alkanes where some of the hydrogen atoms are replaced by fluorine atoms. In one embodiment, the semifluorinated alkane (which may be abbreviated as SFAs) described and used according to the present disclosure comprises of one linear non-fluorinated hydrocarbon segment and of one linear perfluorinated hydrocarbon segment with the perfluorinated hydrocarbon segment attached to the nonfluorinated hydrocarbon segment In a preferred embodiment, the semifluorinated alkanes used in the context of the present disclosure are preferably liquid semifluorinated alkanes.

[0019] In one embodiment, the semifluorinated alkanes have the chemical formula F(CF2)n(CH2)mH, wherein n and m are integers defining the number of carbons in the perfluorinated hydrocarbon segment and non-fluorinated hydrocarbon segment respectively. In some embodiments, the semifluorinated alkane featured in the compositions according to the present disclosure is a semifluorinated alkane of formula F(CF2)n(CH2)mH, wherein n is an integer selected from 4 to 6 and m is an integer selected from 4 to 10. In a further embodiment, the semifluorinated alkanes is a semifluorinated alkane of formula F(CF2)n(CH2)mH, wherein n is an integer selected from 4 to 6 and m is an integer selected from 4 to 8.

[0020] A nomenclature which is also frequently used for linear semifluorinated alkanes designates the perfluorinated hydrocarbon segment as Fn and a non- fluorinated segment as Hm, i.e. FnHm, wherein F means a perfluorinated hydrocarbon segment, H means a non-fluorinated segment, and n and m define the number of carbon atoms of the respective segment For example, F3H3 is used for perfluoropropylpropane, F(CF2)3(CH2)3H. This type of nomenclature is usually used for compounds having linear i.e. unbranched segments.

[0021] Therefore, unless otherwise indicated, it should be assumed that F3H3 means 1-perfluoropropylpropane, rather than its structural isomers e.g. branched isomers such as 2-perfluoropropylpropane, 1 -perfluoroisopropylpropane or 2 -perfluoroisopropylpropane.

[0022] In one embodiment of the present disclosure, the composition may comprise a semifluorinated alkane selected from F4H5, F4H6, F4H8, F6H4, F6H8, F6H10. In another embodiment, the composition comprises a semifluorinated alkane selected from the group consisting of F4H5, F4H6, F4H8, F6H4, F6H8, F6H10, their structural isomers, and mixtures thereof. The chemical formula of F4H5, F4H6, F4H8, F6H4, F6H8, F6H10 maybe expressed respectively as F(CF2)4(CH2)5H, F(CF2)4(CH2)6H, F(CF2)4(CH2)8H, F(CF2)6(CH2)4H, F(CF2)6(CH2)8H and F(CF2)6(CH2)10H. In another embodiment, the composition comprises a semifluorinated alkane selected from F4H5, F6H8, F4H8, or their structural isomers, and any mixture of these semifluorinated alkanes.

[0023] In some embodiments, the semifluorinated alkane featured in the composition according to the present disclosure is F4H5 (1-perfluorobutyl-pentane) or F6H8 (1 -perfluorohexyl-octane), their structure isomers or any mixtures thereof. F4H5, or 1-perfluorobutyl-pentane, with the chemical formula F(CF2)4(CH2)5H, is an inert, water-insoluble liquid, with a density of 1.284 g / cm3at 25 °C and refractive index of 1.3204 at 20 °C. Alternative nomenclature for this compound includes F4H5, wherein F denotes a linear perfluorinated alkane segment comprising 4 carbon atoms and wherein H denotes a linear and non-fluorinated alkane segment of 5 carbon atoms. The semifluorinated alkane, F6H8, where F denotes a linear perfluorinated alkane segment comprising 6 carbons and H denotes a linear and non-fluorinated alkane segment of 8 carbon atoms, is also known as 1-perfluorohexyl-octane and has the chemical formula F(CF2)6(CH2)8H. F6H8 is also a chemically and physiologically inert, water-insoluble liquid, and has a density of 1.35 g / cm3 at 25 °C and refractive index of 1.3432 at 20 °C. In one embodiment, the liquid pharmaceutical composition comprises, or essentially consists of cyclosporine dissolved in F6H8, or dissolved in a mixture of F6H8 and a co-solvent, wherein the co-solvent is such as described herein.

[0024] In an optional embodiment, the composition may comprise more than one semifluorinated alkane. In one embodiment, the composition comprises 1-perfluorohexyl-octane and 2-per fluorohexyl- octane, optionally wherein the 2 -perfluorohexyl-octane is present in an amount of up to 2 % (w / w), or up to 1 % (w / w), up to 0.5 % (w / w), or up to 0.2 % (w / w); or in an amount of 0.1 % to 2% (w / w), or 0.01% to 1 % (w / w) or 0.01% to 0.5 % (w / w) or 0.5% to 5 % (w / w) in respect total weight of the mixture of the two semifluorinated alkanes. In one embodiment, the composition comprises 1-perfluorohexyl-octane and 2-perfluorohexyl-octane, optionally wherein the 2 -perfluorohexyl-octane is present in an amount of up to 2 % (v / v), or up to 1 % (v / v), up to 0.5 % (v / v), or up to 0.2 % (v / v); or in an amount of 0.1 % to 2% (v / v), or 0.01% to 1 % (v / v) or 0.01% to 0.5 % (v / v) or 0.5% to 5 % (v / v) in respect total volume of the mixture of the two semifluorinated alkanes.

[0025] In some embodiments, the composition according to the present disclosure essentially consists of cyclosporine, and at least one of the semifluorinated alkanes as described herein. In other words, there are no further excipients such as the co-solvent present For example, in one embodiment, the composition consists of cyclosporine and a semifluorinated alkane selected from the group consisting of F4H5, F4H6, F4H8, F6H4, F6H8, F6H10, or their structural isomers and any combinations thereof.

[0026] In other embodiments, the composition according to the present disclosure essentially consists of cyclosporine, a semifluorinated alkane and optionally, a co-solvent.

[0027] As used herein, the term "consists” and related terms "consisting” or "consist” is to be understood as meaning that no other features, other than those prefaced by the term are present. In the context of compositions of the present disclosure, if any other constituent or component is present in the composition other than those prefaced by such term, then it is present only in trace or residual amounts such as to confer no technical advantage or relevance in respect of the object of the disclosure, such as may be further understood by the term ‘essentially” or "substantially” used in conjunction with these terms (e.g. ‘essentially consisting of’). In contrast, the term ‘comprising” or related terms "comprises” or "comprise” in the context of compositions, is to be understood as meaning that other features, other than those prefaced by the term may be present in the composition. Also as used herein, the term ‘a’ or ‘an’, or ‘the’ in relation to a semifluorinated alkane, or the co-solvent or any other constituent part of the composition does not exclude the plurality unless the context clearly indicates or requires otherwise. The terms ‘a’, ‘an’ and ‘the’ may be understood to correspond to ‘at least one’ or as ‘one or more’ unless defined otherwise, or unless the context clearly indicates otherwise.

[0028] In one embodiment of the present disclosure, the amount of semifluorinated alkane (or mixture of semifluorinated alkanes) in the composition is in an amount of at least 50 to 90% (w / w), or 85 to 95% (w / w), or 87 to 92% (w / w), or 88 to 91% (w / w) or 89 to 90% (w / w)„ with respect to the total weight of the composition. In some embodiments, the amount of semifluorinated alkane in the composition is in an amount of at least 50%, 60%, 70%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92% or at least 95%, 96%, 97%, 98%, or at least 99% (w / w) with respect to the total weight of the composition.

[0029] In other embodiments according to the present disclosure, the pharmaceutical composition comprises from about 80% (w / w) to about 99% (w / w), or from about 87% (w / w) to about 92% (w / w), or from about 88 % (w / w) to about 91% (w / w), or from about 89 % (w / w) to about 90% (w / w)or from about 90 % (w / w) to about 99.9% (w / w) of a semifluorinated alkane as based on the total weight of the composition.

[0030] In another embodiment, the pharmaceutical composition comprises at least about 50% to 99 % (w / w), or at least 87% to 92% (w / w), or 88 % (w / w) to 91% (w / w), or 89 % (w / w) to 90% (w / w) or at least about 90 to 95%, 96%, 97%, 98%, or 99% (w / w) of 1 -perfluorohexyl-octane (F6H8), and optionally 2-perfluorohexyl-octane, based on the total weight of the pharmaceutical composition. In a further embodiment, the pharmaceutical composition comprises at least about 90% to 99% (w / w), or at least or at least 87% to 92% (w / w), or at least 88 % (w / w) to 91% (w / w), or at least 89 % (w / w) to 90% (w / w) of 1-perfluorohexyl-octane (F6H8), and up to about 0.2%, 0.3%, 0.4%, 0.5% (w / w) or up to about 1 % (w / w) of 2-perfluorohexyl-octane, based on the total weight of the pharmaceutical composition.

[0031] In one embodiment of the present disclosure, the amount of semifluorinated alkane (or mixture of semifluorinated alkanes) in the composition is in an amount of at least 50 to 90% (v / v), or 85 to 95% (v / v), or 87 to 92% (v / v), or 88 to 91% (v / v) or 89 to 90% (v / v)„ with respect to the total weight of the composition. In some embodiments, the amount of semifluorinated alkane in the composition is in an amount of at least 50%, 60%, 70%, 80%, 85%, 87%, 88%, 89%, 90%, 91%, 92% or at least 95%, 96%, 97%, 98%, or at least 99% (v / v) with respect to the total weight of the composition. In other embodiments according to the present disclosure, the pharmaceutical composition comprises from about 80% (v / v) to about 99% (v / v), or from about 87% (v / v) to about 92% (v / v), or from about 88 % (v / v) to about 91% (v / v), or from about 89 % (v / v) to about 90% (v / v)or from about 90 % (v / v) to about 99.9% (v / v) of a semifluorinated alkane as based on the total weight of the composition.

[0032] In another embodiment, the pharmaceutical composition comprises at least about 50% to 99 % (v / v), or at least 87% to 92% (v / v), or 88 % (v / v) to 91% (v / v), or 89 % (v / v) to 90% (v / v) or at least about 90 to 95%, 96%, 97%, 98%, or 99% (v / v) of 1-perfluorohexyl-octane (F6H8), and optionally 2-perfluorohexyl-octane, based on the total weight of the pharmaceutical composition. In a further embodiment, the pharmaceutical composition comprises at least about 90% to 99% (v / v), or at least or at least 87% to 92% (v / v), or at least 88 % (v / v) to 91% (v / v), or at least 89 % (v / v) to 90% (v / v) of 1-perfluorohexyl-octane (F6H8), and up to about 0.2%, 0.3%, 0.4%, 0.5% (v / v) or up to about 1 % (v / v) of 2-per fluorohexyl- octane, based on the total weight of the pharmaceutical composition.

[0033] The liquid pharmaceutical composition of the present disclosure is a solution, preferably a clear solution. The term "solution” may be used interchangeably with the term ‘a clear solution” and as understood herein, refers to a liquid solution in which all solutes (e.g. cyclosporine) are fully dissolvable or dissolved under room temperature conditions i.e. between 15 and 25 °C. The clear solution does not comprise of any particulate or solid phase components and preferably has a refractive index approximate to that of water (i.e. 1.333) or else a refractive index proximal to that of the solvent or mixture of solvents at room temperature. The liquid pharmaceutical compositions according to the present disclosure are not emulsions or are not suspensions.

[0034] The compositions according to the present disclosure may optionally comprise a co-solvent A co-solvent as understood herein is a compound which is suitable for enhancing the solubility of or can assist in solubilizing cyclosporine and is a preferably a liquid that is physiologically compatible for application to the skin and / or hair follicles, and moreover, is a compound that is fully miscible with the semifluorinated alkane. That is, the co-solvent mixes with the semifluorinated alkane to form a coherent and single phase, or a solution that may be used then to solvate or dissolve cyclosporine. In an optional embodiment, the co-solvent may be a compound which is first dissolved in the semifluorinated alkane, but which in combination with the semifluorinated alkane may act as a solvent for cyclosporine and / or optionally any other solute excipients which may be comprised in the composition. In some embodiments, the composition for use in topical treatment of according to the present disclosure comprises a co-solvent. The co-solvent may be an alcohol, such as an alkyl alcohol. In some embodiment, the alcohol is selected from ethanol and isopropanol. In other embodiments, the co-solvent is selected from the group consisting of ethanol, isopropanol, N-methyl pyrrolidone, dimethylformamide, dimethylsulfoxide and combinations thereof. In further embodiments, the co-solvent is selected from isopropanol, N-methyl pyrrolidone and combinations thereof.

[0035] In some embodiments, the composition for use according to the present disclosure comprises cyclosporine dissolved in a mixture of a semifluorinated alkane and a co-solvent. In some embodiments, the co-solvent is present in the composition in an amount of 1 to 20 % (w / w), or of 5 to 20 % (w / w), or of 5 to 10 % (w / w), based on the total weight of the composition. In other embodiments, the amount of co-solvent in the composition is present in the composition in an amount of 1 to 20 % (v / v), or of 5 to 20 % (v / v), or of 5 to 10 % (v / v), based on the total volume of the composition.

[0036] In some embodiments, the co- solvent is isopropanol, in an amount of 9% to 11% (w / w), or in an amount of 10% (w / w) relative to the total weight of the composition, or in an amount of 8 to 12 % (v / v), or in an amount of 9% to 11% (v / v), or in an amount of 10% (v / v) relative to the total volume of the composition. In other embodiments, the co-solvent is N-methyl pyrrolidone in an amount of 5 % (w / w) relative to the total weight of the composition.

[0037] As understood herein, the term "% (w / v)" unless otherwise indicated refers to the amount of a component of a composition as a weight percentage in relation to the total volume of the composition (with "w” denoting weight and "v” denoting volume). For example, a 0.1 % (w / v) would correspond to 1.0 mg of a component in 1 mL of the composition. The term "% (w / w)” or alternatively, "wt %" as used herein and unless otherwise indicated refers to the amount of a component of a composition as a weight percentage in relation to the total weight of the composition with ‘w’ denoting weight The term % (v / v), or volume percent as used herein, and unless otherwise indicated, refers to the volume of a component of a composition in relation to the total volume of the composition.

[0038] As understood herein, the term ‘essentially free’, or alternatively ‘substantially free’ or ‘component- free’ in reference to a composition constituent or component refers to the presence of said component in no more than trace amounts and that if present in trace amounts the component does not provide any technical contribution or material advantage to the composition. The compositions described herein are, in one embodiment, essentially free of water. In another embodiment, the composition as described herein may be essentially free of a preservative, such as an anti-microbial preservative. In other embodiments, the composition as described herein are essentially free of a surfactant

[0039] The pharmaceutical composition for use according to the embodiments of the present disclosure is preferably a liquid with low viscosity. In some embodiments, the composition has a dynamic viscosity at room temperature of between 1 and 4 mPas. In some embodiments, the composition is a sprayable composition, allowing for specific targeting or localized application of the composition to well-defined or select areas of the skin. Preferably, the compositions according to the present disclosure may have advantageous over other topical formulations of cyclosporine in oil with respect to improved compliance, or tolerance of applying the composition to the skin. Oil-based formulations may for example be greasy, can lead to contamination of clothing or other objects, and may as such be inconvenient for the subject or patient.

[0040] The term ‘about’ as used herein and in reference or connection to a parameter, for example such as the amount or concentration of a compound dissolved or suspended in the composition includes the precise value as defined, as well as any value falling within the degree of variability usually observed in measuring or determining these parameters using the standard techniques and equipment known in the art and field.

[0041] In some embodiments, the composition for use according to the present disclosure comprises or consists of cyclosporine dissolved in 1 -perfluorohexyloctane and isopropanol, and optionally 2 -perfluorohexyloctane.

[0042] Preferably, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2 -perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2-perfluorohexyloctane; or the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (w / w) isopropanol and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0043] It is further preferred that the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2 -perfluorohexyloctane; or the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (w / w) isopropanol and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0044] Preferably, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2-perfluorohexyloctane; or the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (w / w) isopropanol and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0045] In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87 to 92% (w / w) 1-perfluorohexyloctane, 8 to 12% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87.5% (w / w) to 92% (w / w) 1-perfluorohexyloctane, 8 to 12 % (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2-perfluorohexyloctane; or the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87.7% (w / w) to 92% (w / w) 1-perfluorohexyloctane, 8 to 12% (w / w) isopropanol, and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0046] It is preferred that the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88 to 91% (w / w) 1-perfluorohexyloctane, 9 to 11% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88.5 to 91% (w / w) 1-perfluorohexyloctane, 9 to 11 % (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2-perfluorohexyloctane; or the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88.7% (w / w) to 91% (w / w) 1-perfluorohexyloctane, 9 to 11% (w / w) isopropanol, and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0047] It is preferred that the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89 to 90% (w / w) 1-perfluorohexyloctane, 10% (w / w) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (w / w) 2 -perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89.5 to 90% (w / w) 1-perfluorohexyloctane, 10 % (w / w) isopropanol, and optionally up to about 0.5% (w / w) 2 -perfluorohexyloctane; or the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89.7% (w / w) to 90% (w / w) 1-perfluorohexyloctane, 10% (w / w) isopropanol, and optionally up to about 0.3% (w / w) 2-perfluorohexyloctane.

[0048] Preferably, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2-perfluorohexyloctane; or the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12% (v / v) isopropanol and optionally up to about 0.3% (v / v) 2-perfluorohexyloctane.

[0049] It is further preferred that the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2-perfluorohexyloctane; or the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 9 to 11% (v / v) isopropanol and optionally up to about 0.3% (v / v) 2-perfluorohexyloctane.

[0050] Preferably, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2-perfluorohexyloctane; or the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 10% (v / v) isopropanol and optionally up to about 0.3% (v / v) 2-perfluorohexyloctane.

[0051] In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87 to 92% (v / v) 1-perfluorohexyloctane, 8 to 12% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87.5% (v / v) to 92% (v / v) 1 -perfluorohexyloctane, 8 to 12 % (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2 -perfluorohexyloctane; or the composition for use comprises or consists of 8 to 12 mg / ml cyclosporine dissolved in 87.7% (v / v) to 92% (v / v) 1 -perfluorohexyloctane, 8 to 12% (v / v) isopropanol, and optionally up to about 0.3% (v / v) 2 -perfluorohexyloctane.

[0052] It is preferred that the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88 to 91% (v / v) 1-perfluorohexyloctane, 9 to 11% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88.5 to 91% (v / v) 1-perfluorohexyloctane, 9 to 11 % (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2 -perfluorohexyloctane; or the composition for use comprises or consists of 9 to 11 mg / ml cyclosporine dissolved in 88.7% (v / v) to 91% (v / v) 1-perfluorohexyloctane, 9 to 11% (v / v) isopropanol, and optionally up to about 0.3% (v / v) 2-perfluorohexyloctane.

[0053] It is preferred that the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89 to 90% (v / v) 1-perfluorohexyloctane, 10% (v / v) isopropanol, and optionally up to about 0.3%, 0.5%, or up to about 1% (v / v) 2-perfluorohexyloctane. In some embodiments, the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89.5 to 90% (v / v) 1-perfluorohexyloctane, 10 % (v / v) isopropanol, and optionally up to about 0.5% (v / v) 2-perfluorohexyloctane; or the composition for use comprises or consists of 10 mg / ml cyclosporine dissolved in 89.7% (v / v) to 90% (v / v) 1-perfluorohexyloctane, 10% (v / v) isopropanol, and optionally up to about 0.3% (v / v) 2-perfluorohexyloctane.

[0054] As discussed herein, the compositions according to the present disclosure may be used for the topical treatment of alopecia areata. As understood and used herein, the term ‘alopecia areata’ generally refers to the disease as featured or present on an area, or combination of individual areas of skin with respect to any part or multiple parts of a subject’s anatomy that may be affected by the disease.

[0055] The topical treatment of alopecia areata is provided by topical application of a composition as described herein onto the skin of a subject in need thereof. The subject to be treated is preferably a human, but in some embodiments may also be a veterinary subject The composition according to the present disclosure may be topically administered to the skin via a number of methods insofar as the composition may be applied directly onto the surface of the skin where the treatment is needed.

[0056] The subject to be treated may have an area of their skin that is afflicted with alopecia areata, or afflicted with hair loss, and / or in need of hair growth, e.g. as a result of alopecia areata, to which the composition may be applied. In some embodiments, the area is an individual area, or in other words a well-defined or localized area of their skin anatomy where alopecia areata, hair loss or the need for hair growth is present, e.g. such as a patch of skin. In some embodiments, the subject has multiple individual areas of their skin which are afflicted with alopecia areata, hair loss or the need for hair growth, to which the composition is applied.

[0057] In some embodiments, the composition is topically applied to one or more individual areas of the skin (e.g. one, or plurality of individual areas of the skin), wherein the surface area of the individual areas of the skin independently do not exceed about 500 cm2, or does not exceed about 400, 300, 200, 100, 50 or 25 cm2. The shape of said individual areas maybe of any shape.

[0058] Alopecia areata may be classified into different sub-types depending on the clinical presentation or pattern of the hair loss with respect to the subject’s anatomy such as the head (e.g. the scalp), or specific areas or the whole of the body. Sub-types may include, but not be limited to patchy alopecia areata, alopecia areata totalis, ophiasis, diffuse alopecia areata and alopecia areata totalis. Patchy alopecia areata is characterized by at least one, or multiple patches of hair loss, optionally contiguous patches of hair loss; the patches are typically well-defined or may be round or oval in shape. Alopecia areata totalis refers to essentially complete or near complete loss of hair from the scalp of a subject Alopecia areata universalis may refer to essentially complete, or near complete loss of hair from the skin of all haircomprising parts of a subject’s body. Ophiasis refers to hair loss in a band shape, usually circumferencing at least one part of the head. Diffuse alopecia areata refers to the disease manifestation where loss of hair can be clinically determined, but where there are no distinct or discrete bald patches or bands, or individualized areas of hair loss. The compositions according to the present disclosure may be used for the topical treatment of any one of these types of alopecia areata.

[0059] In some embodiments, the composition is used for the topical treatment of alopecia areata, hair loss, and or for use in the topical stimulation of hair growth of one or more (individual) areas of the skin on the head or scalp of a subject Areas, or individual areas of the skin comprising hair-follicles on the head may include the face, beard, eyebrows, eyelashes, and the scalp. In some embodiments, the composition may be independently topically administered to one or more individual areas of the skin of the head of the subject selected from the face, the beard, eyebrows, eyelashes, and the scalp. In other embodiments, the composition according to the present disclosure is provided for topical administration to the scalp, i.e. for the topical treatment of alopecia areata, hair loss or for topical stimulation of hair growth on the scalp. In one embodiment, the composition is not used for the topical stimulation of hair growth of the eyelashes of a subject and / or is not used for the topical treatment of hypotrichosis of the eyelashes of a subject

[0060] The onset of alopecia areata, or hair loss may be triggered by a number of factors. In certain embodiments, the composition is used for in the treatment of a subject suffering from alopecia areata, or for the treatment of a subject suffering from hair loss, wherein the alopecia areata or hair loss is related to, or caused by chemotherapy.

[0061] Preferably, the composition is topically applied, or applicable to the skin without contact with the subject’s hand or fingers, during and / or after application of the composition to the skin or skin area. The composition may be applied using a relevant dispensing means, as will further be discussed below, directly onto the skin or targeted area of the skin requiring treatment, without the need for the composition to be massaged, or rubbed into the skin. Or in other words, the composition may, in some embodiments, be rapidly adsorbed onto the skin.

[0062] Advantageously, this may help a subject for example avoid, or reduce the risk of unnecessary contact or transfer of the composition and cyclosporine as an active ingredient to other parts of the body, or areas skin not requiring the treatment

[0063] In some embodiments, the composition according to the present disclosure is effective in adsorbing into the hair follicles and / or is effective in accumulating into hair follicles and / or associated sebaceous glands.

[0064] In a further and related aspect, the present disclosure relates to a liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally, a co-solvent, for use topically in the treatment of hair loss. The composition may be used i.e. applied topically to a hair-follicle comprising area of skin, such as described herein above which is generally afflicted with a loss of hair. In some embodiments, the hair loss is caused by alopecia areata. In other embodiments, the hair loss may be caused by other factors resulting in the degradation of the hair shaft of the hair follicle, or breakage of hair.

[0065] Preferably, the loss of hair is not permanent but is reversible, such that the application of the composition may act to prevent further, or additional hair loss, and / or to promote hair growth.

[0066] In this respect, in yet a further and related aspect, the present disclosure also relates to a liquid pharmaceutical composition for topical use in stimulating hair growth wherein the composition comprises cyclosporine dissolved in a semifluorinated alkane and optionally, a co-solvent such as described in any of the embodiments or combination of embodiments herein. In some embodiments, the liquid pharmaceutical composition for topical use in stimulating hair growth and optionally causing hypertrichosis, is topically administered to subjects suffering from hair loss and / or with abnormal hair pattern.

[0067] The compositions as described herein are preferably used for the treatment of a mammalian, or more preferably, a human subject. In some embodiments, the subject may be a veterinary subject.

[0068] The use of a pharmaceutical composition as described in any one of the above embodiments in the manufacture or preparation of a medicament or a medicine for the treatment of a subject in need thereof in relation to any one of the disease or medical conditions described herein is also provided for in the context of the present disclosure. Further provided for are also methods of treating subjects with, or suffering from any of the medical conditions or disorders described herein; or methods of preventing or ameliorating medical conditions or disorders in said subjects, wherein said methods may comprise a step of topically administering a composition as described herein to the skin of a subject in need thereof.

[0069] In yet another aspect, the present disclosure relates to a kit comprising a composition according to any one or combination of embodiments described herein, a container adapted for holding the composition and optionally a means for dispensing the composition and / or instructions for use, wherein the instructions for use comprise any one of the uses or methods of treatment as described herein.

[0070] In some embodiments of the kit, the kit comprises a means for dispensing the composition. Said means may be affixed or reversibly affixable to the container. In some embodiments, the dispensing means is a sponge applicator, or a spray applicator, e.g. a means for producing and / or targeting a spray or fine mist of the composition to the intended application area of the skin. In some embodiments, the container comprised in the kit is a single use container. In other, and more preferred embodiments, the container is a multi-use container. The disclosure further comprises the following items:

[0071] 1. A liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally a co-solvent, for use in the topical treatment of alopecia areata.

[0072] 2. A liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally a co-solvent, for use in the topical treatment of hairloss.

[0073] 3. A liquid pharmaceutical composition comprising cyclosporine dissolved in a semifluorinated alkane and optionally a co-solvent, for use in the topical stimulation of hair growth.

[0074] 4. The composition for use according to any preceding item, wherein the concentration of cyclosporine in the composition is 1 to 30 mg / ml, 1 to 25 mg / ml, 1 to 10 mg / ml, or 1 to 5 mg / ml, 5 to 20 mg / ml, or 3 to 15 mg / ml.

[0075] 5. The composition for use according to any preceding item, wherein composition comprises 50 to 99 % (w / w) of the semifluorinated alkane based on the total weight of the composition.

[0076] 6. The composition for use according to any preceding item, wherein the semifluorinated alkane is a semifluorinated alkane of formula F(CF2)n(CH2)mH, wherein n is an integer selected from 4 to 6 and m is an integer selected from 4 to 10.

[0077] 7. The composition for use according to any preceding item, wherein the semifluorinated alkane is selected from the group consisting of F4H5, F4H6, F4H8, F6H6, F6H4, F6H8 and F6H10; or is selected from the group consisting of F4H5, F6H8 and F4H8; or wherein the semifluorinated alkane is F6H8.

[0078] 8. The composition for use according to any preceding item, wherein the co-solvent is selected from the group consisting of ethanol, isopropanol, N-methyl pyrrolidone, dimethylformamide, dimethylsulfoxide and combinations thereof; or wherein the cosolvent is selected from isopropanol, N-methyl pyrrolidone and combinations thereof.

[0079] 9. The composition for use according to any preceding item, wherein the co-solvent is present in the composition in an amount of 1 to 20 % (w / w) based on the total weight of the composition.

[0080] 10. The composition for use according to any preceding item, wherein the co-solvent is: a) isopropanol, in an amount of 10% (w / w) relative to the total weight of the composition; or

[0081] b) N-methyl pyrrolidone, in an amount of 5 % (w / w) relative to the total weight of the composition.

[0082] The composition for use according to any preceding item, wherein the composition is in the form of a clear solution.

[0083] The composition for use according to any preceding item, wherein the composition essentially consists of cyclosporine, the semifluorinated alkane and optionally, the cosolvent

[0084] The composition for use according to any preceding item, wherein the composition has a dynamic viscosity at room temperature of between 1 and 4 mPas.

[0085] The composition for use according to any preceding item, wherein the composition is topically administered to the skin of a subject in need thereof.

[0086] The composition for use according to item 14, wherein the composition is topically administered to an area of skin of a subject that is afflicted with alopecia areata or hair loss, and / or in need of hair growth.

[0087] The composition for use according to any preceding item, wherein the composition is applied to one or more individual areas of the skin, wherein the total surface area of said areas of the skin independently do not exceed about 500 cm2, or does not exceed about 400, 300, 200, 100, 50 or 25 cm2.

[0088] The composition for use according to any preceding item, wherein the composition is independently topically administered to one or more individual areas of the skin on the head or the scalp of a subject

[0089] The composition for use according to item 16 or 17, wherein the individual area of the skin comprises hair follicles; optionally wherein the individual area of the skin of the subject’s head is selected from the face, beard, scalp, eyebrows, and eyelashes.

[0090] The composition for use according to any preceding item, wherein the composition is effective in adsorbing into the hair follicles and / or is effective in accumulating into hair follicles and / or associated sebaceous glands.

[0091] The composition for use according to any preceding item, wherein the composition is topically applied, or applicable to the skin without contact with the subject’s hand or fingers during or after application of the composition to the skin or skin area. 21. The composition for use according to any preceding item, wherein the composition is for use in the treatment of a subject suffering from alopecia areata or hair loss, wherein the alopecia areata or hair loss is related to, or caused by chemotherapy. 22. A kit comprising a composition, or composition for use as described in any preceding item, a container adapted for holding the composition and optionally a means for dispensing the composition, and / or instructions for use.

[0092] 23. The kit according to item 22, wherein the dispensing means is a sponge applicator or a spray applicator.

[0093] 24. The kit according to item 22 to 23, wherein the container is a single-use container, or a multi-use container.

[0094] The following examples serve to illustrate the invention, however, should not be understood as restricting the scope of the invention.

[0095] EXAMPLES

[0096] Example 1 - Preparation of compositions for use in the treatment of alopecia areata

[0097] Solutions comprising cyclosporine in a semifluorinated alkane are prepared by dissolving the cyclosporine in the semifluorinated alkane, and where a co-solvent is used, by dissolving the cyclosporine in a combination of the semifluorinated alkane and the co-solvent If a co-solvent is used, the solution is prepared by a step of first mixing the semifluorinated alkane with the co-solvent followed by a second step of dissolving the cyclosporine in said mixture. The following compositions are prepared according to these methods:

[0098] Table 1

[0099] Cyclosporine concentration Semifluorinated Co-solvent

[0100] alkane

[0101] 10 mg / ml F6H8 10 wt% isopropanol

[0102] 10 mg / ml F6H8 10 wt% isopropanol

[0103] 25 mg / ml F6H8 5 wt% N- methyl pyrrolidone (NMP) 25 mg / ml F6H8 5 wt% N- methyl pyrrolidone (NMP)

[0104]

[0105] 10 mg / ml F4H5 10 wt% isopropanol 10 mg / ml F4H5 10 wt% isopropanol

[0106] 25 mg / ml F4H5 5 wt% N- methyl pyrrolidone (NMP) 25 mg / ml F4H5 5 wt% N- methyl pyrrolidone (NMP) 10 mg / ml F4H8 10 wt% isopropanol

[0107] 25 mg / ml F4H8 5 wt% N- methyl pyrrolidone (NMP) 1 mg / ml F6H8 —

[0108] 1 mg / ml F4H5 —

[0109]

[0110] Example 2 - Experimental mouse model of alopecia areata

[0111] Experimental alopecia areata was induced in C3H / HeJ mice by skin grafting (Gund R, Christiano AM. Impaired autophagy promotes hair loss in the C3H / HeJ mouse model of alopecia areata. Autophagy. 2023 Jan;19(l):296-305). In brief, female donor mice with chronic alopecia areata were euthanized, and full-thickness skin grafts of ~2 cm in diameter from donor mice were grafted onto the back of 10- week old normal-haired female recipient C3H / HeJ mice. Hair loss typically began at around 4-6 weeks after grafting. Mice that were sham grafted were used as controls. ImageJ software was used to quantify % area of skin exhibiting hair loss. Briefly, area of skin without hair shafts is measured and normalized to total body area. The area under the curve (AUC) was measured for line curves obtained by measuring % area showing hair loss over time after grafting.

[0112] Grafted alopecia areata mice were treated with the test substance starting 6 weeks after grafting for their ability to reverse the hair loss in the area of the grafted areata skin. The test substances included:

[0113] (A) verum: 10 mg / mL cyclosporine dissolved in 1 -perfluorohexyloctane (F6H8) and 10 % (v / v) isopropanol

[0114] (B) control (Vehicle): 1 -perfluorohexyloctane (F6H8)

[0115] The test substances were applied topically over a period of six months as a spray to the grafted areata skin, which was about 15 cm2.

[0116] Result: The verum samples (A) demonstrated re-growth of hair in the area of the grafted areata skin in the mouse model starting after three months of treatment, with full re-growth of hair after 6 months. In contrast, no growth was observed with the control samples (B), as shown when comparing % of re-growth over the treatment period.

Claims

DRM23P03PC2Claims1. A liquid pharmaceutical composition comprising 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane and 8 to 12 % (v / v) isopropanol, for use in the topical treatment of alopecia areata.

2. A liquid pharmaceutical composition comprising 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane and 8 to 12 % (v / v) isopropanol, for use in the topical treatment of hair loss.

3. A liquid pharmaceutical composition comprising 8 to 12 mg / ml cyclosporine dissolved in 1-perfluorohexyloctane, 8 to 12 % (v / v) isopropanol, for use in the topical stimulation of hair growth.

4. The composition for use according to any preceding claim, wherein the concentration of cyclosporine in the composition is 9 to 11 mg / ml or about 10 mg / ml.

5. The composition for use according to any preceding claim, wherein the composition comprises isopropanol at 9 to 11 % (v / v), or at about 10% (v / v).

6. The composition for use according to any preceding claim, wherein composition comprises up to about 1% (v / v) 2-perfluorohexyloctane.

7. The composition for use according to any preceding claim, wherein the composition is a sprayable composition.

8. The composition for use according to any preceding claim, wherein the composition is in the form of a clear solution.

9. The composition for use according to any preceding claim, wherein the composition essentially consists of cyclosporine, 1-perfluorohexyloctane, isopropanol, and optionally 2-perfluorohexyloctane.

10. The composition for use according to any preceding claim, wherein the composition consists of 8 to 12 mg / ml cyclosporine dissolved in 87.5 to 92% (v / v) 1- perfluorohexyloctane, 8 to 12 % (v / v) isopropanol, and optionally, up to 0.5% (v / v) 2-perfluorohexyloctane.

11. The composition for use according to any preceding claim, wherein the composition consists of 10 mg / ml cyclosporine dissolved in 89.5 to 90% (v / v) 1- perfluorohexyloctane, 10 % (v / v) isopropanol, and optionally, up to 0.5% (v / v) 2- perfluorohexyloctane.

12. The composition for use according to any preceding claim, wherein the composition has a dynamic viscosity at room temperature of between 1 and 4 mPas.

13. The composition for use according to any preceding claim, wherein the composition is topically administered to the skin of a subject in need thereof.

14. The composition for use according to any preceding claim, wherein the composition is topically administered to an area of skin of a subject that is afflicted with alopecia areata or hair loss, and / or in need of hair growth.

15. The composition for use according to any preceding claim, wherein the composition is applied to one or more individual areas of the skin, wherein the total surface area of said areas of the skin independently do not exceed about 500 cm2, or does not exceed about 400, 300, 200, 100, 50 or 25 cm2.

16. The composition for use according to any preceding claim, wherein the composition is independently topically administered to one or more individual areas of the skin on the head or the scalp of a subject17. The composition for use according to claim 15 or 16, wherein the individual area of the skin comprises hair follicles; optionally wherein the individual area of the skin of the subject’s head is selected from the face, beard, scalp, eyebrows, and eyelashes.

18. The composition for use according to any preceding claim, wherein the composition is effective in adsorbing into the hair follicles and / or is effective in accumulating into hair follicles and / or associated sebaceous glands.

19. The composition for use according to any preceding claim, wherein the composition is topically applied, or applicable to the skin without contact with the subject’s hand or fingers during or after application of the composition to the skin or skin area.

20. The composition for use according to any preceding claim, wherein the composition is for use in the treatment of a subject suffering from alopecia areata or hair loss, wherein the alopecia areata or hair loss is related to or caused by chemotherapy.

21. A kit comprising a composition, or composition for use as described in any preceding claim, a container adapted for holding the composition and optionally a means for dispensing the composition, and / or instructions for use.

22. The kit according to claim 21, wherein the dispensing means is a sponge applicator or a spray applicator.

23. The kit according to claim 21 to 22, wherein the container is a single-use container, or a multi-use container.

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