Methods for treating or preventing skin conditions

Abstract

To provide methods and compositions for preventing and treating certain skin disorders caused by UV-induced, age-related, and post-inflammatory cutaneous changes.SOLUTION: A method of treating a dermal disorder in a mammalian subject in need thereof comprises topically administering to the subject a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereoisomer thereof, where the dermal disorder is at least one selected from the group consisting of static wrinkles, fine wrinkles, loss of skin firmness, ephelides, melasma, senile purpura, static rhytides, UV damage, prominent hand veins, field cancerization of epithelium such as in squamous cell carcinoma, keloids, and imbalance in a ratio of collagen isoforms.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 USC § 119(e) to U.S. Provisional Patent Application No. 62 / 744,486, filed October 11, 2018, which is incorporated by reference in its entirety. [Background technology]

[0002] 2. Background of the Invention Several skin disorders result from skin changes induced by environmental factors, hormonal changes, aging, and inflammatory episodes, such as freckles, solar lentigines, melasma, senile purpura, fine lines, static rhytides, prominent hand veins, actinic keratosis and epithelial mass carcinogenesis, keloid formation, imbalance in the ratio of collagen isoforms, etc.

[0003] Ephelides, solar lentigines, melasma, actinic keratosis, and squamous intraepithelial carcinoma are conditions caused by exposure to sunlight. Ephelides are small pigmented lesions commonly known as freckles. Solar lentigines are larger hyperpigmented lesions, while melasma is a larger patchy brown, tan, or blue-gray discoloration of the facial skin. Widespread carcinoma refers to the appearance of actinic damage and epidermal dysplasia over a large area of ​​sun-exposed skin. Actinic keratosis generally appears as pink, scaly papules or plaques, often occurring in areas of widespread carcinoma. If left untreated, widespread carcinoma and actinic keratosis can progress to squamous cell carcinoma.

[0004] Senile purpura, fine lines, static wrinkles, and prominent hand veins are conditions that commonly affect aging skin. Senile purpura is a common benign condition that usually develops after minor trauma and is characterized by the recurring formation of purple ecchymoses (bruises) caused by bleeding into the dermis from fragile capillaries that are easily damaged due to the thinning of the dermis, or scaffolding, of the skin. Static wrinkles are wrinkles that do not change with muscle movement, and prominent hand veins are a condition in which veins become more prominent due to very little body fat or thinning of the structures that support the veins in the hand. Fine lines are the result of sun damage and other external factors that cause the dermis to become irregular and the epidermis to lose its ability to retain moisture.

[0005] Keloids are scars composed primarily of type III (early) or type I (late) collagen. They form from the overgrowth of granulation tissue (type 3 collagen) at the site of a healed cutaneous wound, which is then gradually replaced by type 1 collagen. Keloids are an abnormal scarring response that results in a hard, hyperpigmented plaque whose borders extend beyond the area of ​​primary injury. Keloids disproportionately affect non-whites and are often pruritic, tender, and unsightly. Furthermore, when keloids occur near joints such as the knee or ankle, they can cause stiffness as well as limit range of motion.

[0006] There is a need in the art for compositions and methods that can be used to treat or prevent certain dermal disorders resulting from UV-induced, age-related, and post-inflammatory skin changes. The present invention addresses this need. Summary of the Invention

[0007] In one embodiment, the present invention provides a method for treating and / or preventing dermal disorders in a mammalian subject in need thereof. The method comprises topically administering to the subject a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof. In certain embodiments, the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin tautness, freckles, melasma, senile purpura, static rhytides, prominent hand veins, epithelial mass carcinogenesis such as in squamous cell carcinoma, keloids, and imbalance in collagen isoform ratios.

[0008] In another embodiment, the composition comprises from about 1% to about 0.0001% by weight of rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof. [The present invention 1001] A method for treating a dermal disorder in a mammalian subject in need thereof, comprising the step of topically administering to the subject a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytides, UV damage, prominent hand veins, epithelial mass carcinogenesis such as in squamous cell carcinoma, keloids, and imbalance in the ratio of collagen isoforms. [The present invention 1002] The method of claim 1001, wherein epithelial widespread carcinogenesis includes epidermal dysplasia and actin damage. [The present invention 1003] The method of claim 10, wherein the subject is a human. [The present invention 1004] The method of claim 1001, wherein the composition comprises about 0.1% by weight to about 0.0001% by weight of rapamycin. [The present invention 1005] The method of claim 10, wherein the composition comprises about 0.001% by weight to about 0.0001% by weight of rapamycin. [The present invention 1006] The method of claim 10, wherein said composition further comprises a dermatologically acceptable carrier. [The present invention 1007] The method of the present invention 1006, wherein the dermatologically acceptable carrier is at least one selected from the group consisting of solvents, lubricants, emollients, emulsifiers, moisturizers, thickening waxes, emollients, fragrances, preservatives, and artificial colorants. [The present invention 1008] The method of claim 1006, wherein the dermatologically acceptable carrier comprises petrolatum. [The present invention 1009] The method of claim 10, wherein the composition is applied to the site of a dermal disorder in a subject. [The present invention 1010] A kit for treating and / or preventing a dermal disorder in a mammalian subject in need thereof, comprising a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, an applicator, and instructions for topically administering the composition to the subject, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytides, prominent hand veins, epithelial mass carcinogenesis, keloids, and imbalance in the ratio of collagen isoforms. [Brief description of the drawings]

[0009] For the purpose of illustrating the invention, there are shown in the drawings certain embodiments of the invention. However, the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings. [Figure 1] FIG. 1 shows that rapamycin treatment reduces p16 levels in human skin. [Diagram 2] FIG. 2 shows that biopsies treated with rapamycin demonstrate a reduction in histological markers of aged / damaged skin compared to biopsies treated with placebo. [Diagram 3]FIG. 3 shows that the rapamycin-treated biopsies show a more orderly arrangement of the epithelial layers compared to the placebo-treated biopsies. [Figure 4] FIG. 4 shows that the rapamycin-treated biopsies show a more ordered arrangement of collagen fibers and less solar elastosis compared to the placebo-treated biopsies. [Diagram 5] FIG. 5 shows that rapamycin-treated skin had cytokeratin 5 / 6 staining more focally located in the basal layer of the skin, whereas placebo-treated skin had cytokeratin 5 / 6 staining in the granular layer, suggesting incomplete differentiation that is typical of aged skin. [Figure 6] FIG. 6 shows that in placebo-treated skin, cytokeratin 5 / 6 (CK 5 / 6) staining shows positive staining in the granular layer and extends into the stratum corneum, whereas in rapamycin-treated biopsies, the pattern of CK 5 / 6 staining is more restricted to the basal layer. [Figure 7] Figure 7 shows the clinical improvement of a 67-year-old woman after six months of application of rapamycin cream. For comparison, the placebo-treated left hand is also shown. [Figure 8] FIG. 8 shows that purpura was significantly reduced after 4.5 months of rapamycin treatment. [Figure 9] 9A-9C show that rapamycin protects against UV-induced cell death. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] Detailed Description of the Invention In one aspect, the present invention relates to a method for preventing and / or treating various dermal and epidermal disorders, including, but not limited to, freckles, melasma, senile purpura, static wrinkles, prominent hand veins, epidermal wide-area carcinogenesis, keloid formation, and imbalances in the ratio of collagen isoforms.

[0011] In certain embodiments, the present invention provides compositions comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof. In other embodiments, the compositions comprise rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof, as the only component that exhibits activity against dermal disorders contemplated herein. In yet other embodiments, the compositions comprise rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof, as the only component present in a concentration and / or amount sufficient to exhibit activity against dermal disorders contemplated herein. In yet other embodiments, the compositions are formulated for topical administration.

[0012] Proper differentiation is crucial for skin function, for example, providing a protective layer against water loss, protecting against UV damage from sun exposure, and acting as a barrier against pathogens. These protective functions require orderly differentiation of cells within the epidermis. With aging, differentiation patterns in the skin become disrupted, resulting in a dysfunctional barrier that renders the skin vulnerable to wounds, tears, and / or UV damage. As demonstrated herein, nanomolar concentrations of rapamycin result in more orderly differentiation within the epidermis of human skin while simultaneously reducing markers of aging and cellular dysfunction. p16, a key protein involved in cellular aging in the dermis and epidermis, is downregulated by rapamycin, a cellular ... INK4A The expression of rapamycin is decreased. This protein is associated with age-related changes in the skin (Waaijer, et al., 2016, J Gerontol A Biol Sci Med Sci. 71(8):1022-1028; Waaijer, et al., 2012, Aging Cell 11(4):722-725). Furthermore, histological changes associated with UV damage in the skin are also attenuated by rapamycin treatment. These changes translate into clinical improvements in skin firmness and wrinkles in the elderly.

[0013] definition As used herein, each of the following terms has the meaning associated with it in this section.

[0014] Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. For the most part, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics and chemistry are well known and commonly used in the art.

[0015] The articles (a) and (an) are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0016] The term "about" as used herein will be understood by those of skill in the art and will vary to some extent depending on the context in which it is used. As used herein, the term "about" when referring to a measurable value, such as an amount, duration, and the like, is meant to include a ±20% variation from the particular value, that is, a ±10%, ±5%, ±1%, or ±0.1% variation, such variations being reasonable for performing the disclosed methods.

[0017] As used herein, "dermatologically acceptable carrier" or "dermatologically acceptable excipient" refers to a composition or ingredient suitable for use in contact with human keratinous tissue without undue toxicity, incompatibility, instability, allergic response, and the like.

[0018] A "disease" is a state of health in an animal in which the animal is unable to maintain homeostasis, and if such a disease is not ameliorated, the animal's health will continue to deteriorate. A "disorder" in an animal is a state of health in which the animal is able to maintain homeostasis, but the animal's health is no better than it would be without the disorder. If left untreated, the disorder does not necessarily cause a further deterioration in the animal's health.

[0019] As used herein, the term "effective amount" or "therapeutically effective amount" or "pharmaceutical effective amount" of a compound is used interchangeably to refer to the amount of a compound that is sufficient to produce beneficial effects on the subject to which the compound is administered.As used herein, the term "treat" means to reduce the frequency of symptoms experienced by a patient or subject, or to administer a drug or compound to reduce the severity of symptoms experienced.The appropriate therapeutic amount in each case can be determined by those skilled in the art using routine experimentation.

[0020] As used herein, "instructional material" includes publications, records, diagrams, or any other means of expression that can be used to communicate the usefulness of a compound, composition, assay, or method of the invention in a kit for suppressing or reducing a systemic immune response in a subject. The instructional material of the kit of the invention may, for example, be affixed to a container containing an identified compound, composition, assay, or method of the invention, or may be shipped together with a container containing an identified compound, composition, assay, or method. Alternatively, the instructional material may be shipped separately from the container, with the intention that the recipient will use the instructional material and the compound, composition, assay, or method cooperatively.

[0021] The term "modulate" as used herein means, with respect to a disease state or condition associated with the binding of a compound of the invention to a receptor contemplated by the invention, to directly or indirectly improve or alleviate a disease state or condition that was suboptimal, and in many cases debilitating, even life-threatening, prior to administration of a compound according to the invention. Modulation can occur through agonistic, antagonistic, or mixed agonistic / antagonistic activity (depending on the receptor site).

[0022] As used herein, the term "pharmaceutical acceptable" refers to a relatively non-toxic substance, such as a carrier or diluent, that does not abrogate the biological activity or properties of the composition; i.e., the substance can be administered to an individual without causing undesired biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained.

[0023] The term "pharmaceutical composition" or "composition" as used herein refers to a mixture of at least one compound useful in the present invention with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or excipients. Pharmaceutical compositions facilitate the administration of the compound to an organism. There are many techniques in the art for administering compounds, including, but not limited to, intravenous, oral, aerosol, parenteral, intraocular, pulmonary, intracranial and topical administration. In certain embodiments, administration includes topical administration.

[0024] As used herein, a "subject" refers to a human or non-human mammal. Non-human mammals include farm animals and pets, such as, for example, sheep, cows, pigs, dogs, cats and murines. In certain aspects, the subject is a human.

[0025] As used herein, "topical administration" or "topical application" refers to a medication applied to a body surface such as the skin or mucous membranes.

[0026] The term "treatment" or "treating" as used herein is defined as the application or administration of a therapeutic agent, i.e., a composition useful in the present invention (alone or in combination with another agent), to a subject, or to an isolated tissue or cell line from a subject having a disease or disorder, a symptom of a disease or disorder, or a potential for developing a disease or disorder (e.g., for diagnostic or ex vivo applications) for the purpose of curing, healing, mitigating, alleviating, altering, relieving, improving, enhancing, or affecting the disease or disorder, the symptom of the disease or disorder, or the potential for developing a disease or disorder. Such treatments can be specifically tailored or modified for each individual based on knowledge gained from the field of pharmacogenomics.

[0027] Throughout this disclosure, various aspects of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and should not be construed as an inflexible limitation on the scope of the invention. Thus, the description of a range should be considered to have specifically disclosed all possible subranges as well as individual numerical values ​​within that range. For example, the description of a range such as 1-6 should be considered to have specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc., as well as individual numerical values ​​within that range, such as 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This is true regardless of the breadth of the range.

[0028] composition In one embodiment, the compositions of the invention comprise a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof. Rapamycin is (3S,6R,7E,9R,10R,12R,14S,15E,17E,19E,21S,23S,26R,27R,34aS)-9,10,12,13,14,21,22,23,24,25,26,27,32,33,34,34a-hexadecahydro-9,27-dihydroxy-3-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-[(1R)-2-[(1S,3R,4R)-4-hydroxy-3-[(1R)-2-[(1R)-2-hydroxy-3 ... It is also known as [1,4-oxaazacyclohentriacontin-1,5,11,28,29(4H,6H,31H)-pentone, which has the following structure: TIFF2025024020000001.tif47128

[0029] In certain embodiments, modified forms of rapamycin can be used that improve delivery to specific intracellular compartments or organelles, such as mitochondria, nuclei, lysosomes, and / or the endoplasmic reticulum, etc. Modified forms of rapamycin include ester, amide, ether derivatives, and the like.

[0030] Other mTORC1 inhibitors besides rapamycin are also useful within the methods presented herein to achieve the therapeutic effects described herein. In certain embodiments, the mTORC1 inhibitor is BEZ235, everolimus, AZD8055, temsirolimus, KU-0063794, PI-103, torkinib, tacrolimus, ridaforolimus, INK-128, voxtalisib, Torin-1, omipalisib, OSI-027, PF-04691502, apitolisib, GSK1059615, WYE -354, jedatolicib, AZD-2014, Torin-2, WYE-125132, BGT226, Palomid-529, PP121, WYE-687, CH5132799, Way-600, ETP-46464, GDC-0349, XL388, and zotarolimus, or salts, solvates, enantiomers, or diastereomers thereof. In another embodiment, the mTORC1 inhibitor is at least one selected from the group consisting of rapamycin, ridaforolimus, and everolimus, or salts, solvates, enantiomers, or diastereomers thereof. In yet another embodiment, the mTORC1 inhibitor is rapamycin, or salts, solvates, enantiomers, or diastereomers thereof. A therapeutically effective amount of any of the mTORC1 inhibitors described herein can be from about 0.0001% to about 1%, 0.0005% to about 0.95%, about 0.001% to about 0.85%, 0.002% to about 0.75%, about 0.005% to about 0.5%, about 0.008% to about 0.25%, about 0.01% to about 0.2%, about 0.02% to about 0.15%, about 0.0001% to about 0.001%, about 0.0001% to about 0.01%, or about 0.03% to about 0.1% (w / w) of the composition.A therapeutically effective amount of an mTORC1 inhibitor that can be used in the compositions described herein can be about 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or about 0.0001% (w / w) of the composition.

[0031] In certain embodiments, the therapeutically effective amount of rapamycin in the composition ranges from about 0.0001% to about 1%, 0.0005% to about 0.95%, about 0.001% to about 0.85%, 0.002% to about 0.75%, about 0.005% to about 0.5%, about 0.008% to about 0.25%, about 0.01% to about 0.2%, about 0.02% to about 0.15%, about 0.0001% to about 0.001%, about 0.0001% to about 0.01%, or about 0.03% to about 0.1% (w / w) of the composition. In certain embodiments, the therapeutically effective amount of rapamycin in the composition ranges from about 0.0001% to about 0.001% (w / w). In other embodiments, the therapeutically effective amount by weight of rapamycin in the composition ranges from about 0.0001% to about 0.01% (w / w).

[0032] In certain embodiments, the therapeutically effective amount of rapamycin in the composition is about 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or about 0.0001% (w / w). In certain embodiments, compositions containing 0.001% w / w or less of rapamycin do not inhibit proliferation of dermal cells and maintain the proliferation ability of dermal cells. In certain embodiments, compositions containing 0.001% w / w or less of rapamycin prevent or reduce senescence of dermal cells. In certain embodiments, compositions containing about 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or about 0.0001% (w / w) of rapamycin do not inhibit proliferation of dermal cells, maintain the proliferation ability of dermal cells, and prevent or reduce senescence of dermal cells.

[0033] In certain embodiments, the composition of the present invention further comprises a dermatologically acceptable carrier. The composition of the present invention may comprise about 60% to about 99.9%, alternatively about 70% to about 95%, alternatively about 80% to about 90% of the dermatologically acceptable carrier. In certain embodiments, the dermatologically acceptable carrier is at least one selected from the group consisting of solvents, lubricants, emollients, emulsifiers, moisturizers, thickening waxes, softeners, fragrances, preservatives, and artificial colors. In other embodiments, the dermatologically acceptable carrier is at least one selected from the group consisting of water, fatty alcohols, and volatile organic alcohols. One non-limiting example of a dermatologically acceptable carrier is petrolatum.

[0034] method In one aspect, the present invention is a method for preventing and / or early treatment of UV-induced pigmentation disorders that are the cause of freckles and melasma. In another aspect, the present invention provides a method for preventing and / or treating age-related dermal disorders, such as, but not limited to, senile purpura, static wrinkles, and / or prominent hand veins. In yet another aspect, the present invention provides a method for preventing and / or treating epithelial wide-field carcinogenesis, such as in squamous intraepithelial carcinoma. In yet another aspect, the present invention provides a method for preventing and / or treating keloids. In yet another aspect, the present invention further provides a method for maintaining an optimal ratio of collagen isoforms to prevent aging. In yet another aspect, the present invention further provides a kit for treating or preventing dermal disorders due to skin changes occurring in a mammalian subject in need thereof.

[0035] In certain aspects, the methods comprise topically administering to the subject a therapeutically effective amount of rapamycin, optionally formulated in a skin-tolerable composition.

[0036] In certain embodiments, the composition comprises a therapeutically effective amount of rapamycin. In yet other embodiments, the composition further comprises a dermatologically acceptable carrier. In yet other embodiments, the composition is applied topically to the affected skin area of ​​the subject.

[0037] In certain embodiments, topical formulations of the compositions contemplated herein are used to treat UV-induced, age-related, and post-inflammatory skin changes.

[0038] In certain embodiments, the present invention provides a topical cream comprising a therapeutically effective amount of rapamycin for treating or preventing UV-induced, age-related, and post-inflammatory skin changes.

[0039] In certain embodiments, the skin changes are characterized by the p16 INK4AThe assessment is carried out by measuring aging-associated proteins such as aging-associated markers such as cytokeratin 5 / 6, differentiation markers such as cytokeratin 5 / 6, collagen fiber organization, and clinical measures such as wrinkle severity and skin firmness.

[0040] formulation The relative amounts of active ingredient, dermatologically acceptable carrier, and any additional ingredients in the pharmaceutical compositions of the present invention will vary depending on the identity, size, and condition of the subject being treated. By way of example, the compositions can contain from about 0.0001% to about 1% (w / w) rapamycin. In certain embodiments, the compositions comprise about 0.1%, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or about 0.0001% (w / w) rapamycin.

[0041] In certain embodiments, the therapeutically effective amount by weight of rapamycin in the composition ranges from about 0.0001% to about 1%, 0.0005% to about 0.95%, about 0.001% to about 0.85%, 0.002% to about 0.75%, about 0.005% to about 0.5%, about 0.008% to about 0.25%, about 0.01% to about 0.2%, about 0.02% to about 0.15%, or about 0.03% to about 0.1%.

[0042] Although the description of pharmaceutical compositions provided herein is directed primarily to pharmaceutical compositions suitable for ethical administration to humans, those skilled in the art will understand that such compositions are generally suitable for administration to any type of animal. Modifications of compositions suitable for administration to humans to provide pharmaceutical compositions suitable for administration to a variety of animals are well understood, and veterinary pharmacologists of ordinary skill can design and implement such modifications with no more than routine experimentation, if at all. Subjects to which the administration of the pharmaceutical compositions of the present invention is contemplated include, but are not limited to, humans and other primates, mammals, including commercially relevant mammals such as cows, pigs, horses, sheep, cats and dogs.

[0043] The compositions of the present invention can be administered to a mammal several times per day, or less frequently, such as once per day, once per week, once every two weeks, once per month, or even less frequently, such as once every few months, once per year or less.

[0044] The dosing regimen for administering the compositions of the present invention may be once a day or twice a day. The frequency and concentration of application of the active agent depends on the condition of the skin and the response of the dermis. Application may be continued to achieve the desired effect on the dermis, or the frequency of application may be reduced after satisfactory results are obtained. In certain embodiments, administration requires a minimum of 6-8 weeks to achieve results. Application may be continued beyond the first 6-8 weeks to obtain continued improvement, and once results are achieved, the frequency of application may be reduced. Application may be continued for several years at various levels of application based on the relative severity of the lesion at any one time.

[0045] It will be appreciated that the amount of the composition of the present invention administered per day can be, by way of non-limiting example, administered every day, every other day, every third day, every fourth day, or every fifth day. The frequency of administration will depend on a variety of factors, including, but not limited to, the type and severity of the disease being treated, the type and age of the animal, etc., as will be readily apparent to one of skill in the art.

[0046] In certain embodiments, the compositions of the present invention are formulated with one or more dermatologically acceptable excipients or carriers.In certain embodiments, the pharmaceutical compositions of the present invention contain a therapeutically effective amount of rapamycin and a dermatologically acceptable carrier.Useful dermatologically acceptable carriers include, but are not limited to, glycerol, water, saline, ethanol and other dermatologically acceptable salt solutions, such as phosphates and salts of organic acids.Examples of these and other dermatologically acceptable carriers are described in Remington's Pharmaceutical Sciences (1991, Mack Publication Co., New Jersey).

[0047] The compositions of the present invention may comprise about 0.01% to about 1%, alternatively about 1% to about 10%, alternatively about 10% to about 50% of a dermatologically acceptable carrier. In certain embodiments, the dermatologically acceptable carrier is at least one selected from the group consisting of solvents, lubricants, emollients, emulsifiers, moisturizers, thickening waxes, emollients, fragrances, preservatives, and artificial colors. In other embodiments, the dermatologically acceptable carrier is at least one selected from the group consisting of water, fatty alcohols, and volatile organic alcohols. One non-limiting example of a dermatologically acceptable carrier is petrolatum.

[0048] The carrier may be a solvent or dispersion medium, including, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of a coating agent, such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Prevention of microbial activity can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, sodium chloride, or polyalcohols, such as mannitol, sorbitol, and the like, in the composition. Prolonged absorption of the injectable composition can be achieved by including in the composition an agent that delays absorption, for example, aluminum monostearate or gelatin.

[0049] The preparations may be used in admixture with conventional excipients, i.e., pharma- ceutically acceptable organic or inorganic carrier substances known in the art that are suitable for topical administration.The preparations are sterilized and, if necessary, mixed with auxiliary substances, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts that affect osmotic buffers, colorants, flavorings and / or aromatic substances, etc.They may also be combined with other active agents, such as other analgesics, if necessary.

[0050] As used herein, "additional ingredients" include, but are not limited to, one or more of the following ingredients: excipients; surfactants; dispersing agents; inert diluents; granulating and disintegrating agents; binders; lubricants; coloring agents; preservatives; physiologically degradable compositions, such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents; demulcents; buffers; salts; thickening agents; fillers; emulsifiers; antioxidants; antibiotics; antifungal agents; stabilizers; and pharmaceutically acceptable polymeric or hydrophobic materials. Other "additional ingredients" that can be included in the pharmaceutical compositions of the present invention are known in the art and are described, for example, in Genaro, ed. (1985, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA), which is incorporated herein by reference.

[0051] The composition of the present invention may include a preservative. Preservatives are used to prevent spoilage when exposed to pollutants in the environment. Examples of preservatives useful in the present invention include, but are not limited to, those selected from the group consisting of benzyl alcohol, sorbic acid, parabens, imidurea, and combinations thereof. A particularly preferred preservative is a combination of about 0.5%-2.0% benzyl alcohol and 0.05%-0.5% sorbic acid.

[0052] The composition preferably includes an antioxidant and / or a chelating agent (such as EDTA). Preferred antioxidants for some compounds are BHT, BHA, α-tocopherol and ascorbic acid in the preferred range of about 0.01% to 0.3% of the total weight of the composition, more preferably BHT in the range of 0.03% to 0.1%. Preferably, the chelating agent is present in an amount of 0.01% to 0.5% of the total weight of the composition. Particularly preferred chelating agents include aminopolycarboxylates (e.g., disodium ethylenediaminetetraacetate) and citric acid in the weight range of about 0.01% to 0.20% of the total weight of the composition, more preferably in the range of 0.02% to 0.10%. Chelating agents are useful for chelating metal ions in the composition that may be detrimental to the shelf life of the formulation.

[0053] Topical administration An obstacle to topical administration of pharmaceuticals is the stratum corneum of the epidermis. The stratum corneum is a highly resistant layer composed of proteins, cholesterol, sphingolipids, free fatty acids and various other lipids, and contains keratinocytes and viable cells. One of the factors limiting the penetration rate (flux) of a compound through the stratum corneum is the amount of active that can be loaded or applied to the skin surface. The more active that is applied per unit area of ​​skin, the greater the concentration gradient between the skin surface and the lower layers of the skin, and thus the greater the diffusion force of the active through the skin. Thus, a formulation containing a higher concentration of active is more likely to result in the penetration of more active into the skin at a more consistent rate compared to a formulation containing a lower concentration of active, all other things being equal.

[0054] Preparations suitable for topical administration include, but are not limited to, liquid or semi-liquid preparations, such as liniments, lotions, oil-in-water or water-in-oil emulsions, such as creams, ointments or pastes, and solutions or suspensions.These preparations can be applied directly to the skin, or by using swabs, applicators, spatulas, etc., or in the form of transdermal patches.In certain embodiments, the patch minimizes the loss of medicament due to washing, rubbing, scratching, and / or rubbing of the skin.In other embodiments, the patch enhances the absorption of medicament through the skin, while simultaneously minimizing the exposure of the skin to the medicament.

[0055] Topically administrable formulations contemplated herein can contain, for example, about 0.001% to about 1% (w / w) rapamycin, although the concentration of rapamycin can be as high as its solubility limit in the solvent. In one specific embodiment, the composition contains about 0.0001% to about 0.1% (w / w) rapamycin. Formulations for topical administration can further include one or more additional ingredients described herein.

[0056] Penetration enhancers can be used. These substances increase the rate of drug permeation through the skin. Typical enhancers in the art include ethanol, glycerol monolaurate, PGML (polyethylene glycol monolaurate), dimethyl sulfoxide, etc. Other enhancers include oleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkane carboxylic acids, dimethyl sulfoxide, polar lipids, or N-methyl-2-pyrrolidone.

[0057] One acceptable vehicle for local delivery of some of the compositions of the present invention can include liposomes, the composition of which and their uses are known in the art (e.g., U.S. Patent No. 6,323,219).

[0058] In another embodiment, the topical formulation further comprises other ingredients, such as adjuvants, antioxidants, chelating agents, surfactants, foaming agents, wetting agents, emulsifiers, thickening agents, buffers, preservatives, etc. In another embodiment, a penetration enhancer or permeation enhancer is added to the formulation, which is effective to improve the percutaneous penetration of the active ingredient into and through the stratum corneum, relative to compositions that do not contain a penetration enhancer. A variety of penetration enhancers are known to those skilled in the art, including oleic acid, oleyl alcohol, ethoxydiglycol, laurocapram, alkane carboxylic acids, dimethyl sulfoxide, polar lipids, or N-methyl-2-pyrrolidone. In another aspect, the topical formulation can further comprise a hydrotropic agent, which functions to increase the irregularity of the structure of the stratum corneum, thereby allowing for increased transport through the stratum corneum. A variety of hydrotropic agents, such as isopropyl alcohol, propylene glycol, or sodium xylene sulfonate, are known to those skilled in the art.

[0059] Additional inactive ingredients in topical formulations are well known in the art.These ingredients include, but are not limited to, moisturizers, emollients, pH stabilizers, chelating agents, gelling agents, thickeners, emulsifiers, binders, buffers, carriers, antioxidants, etc.Further examples of such ingredients are included in the Inactive Ingredients for Approved Drugs of the US Food & Drug Administration, available online.Further discussions, as well as the inactive ingredients that can be added to formulations, can be found in "The Science and Practice of Pharmacy", 21st Edition, Lippincott Williams & Wilkins, Philadelphia, Pa. (2006).

[0060] In certain aspects, the gel formulations of the present invention contain inorganic substances such as aluminum salts or organic polymers of natural or synthetic origin to allow for the release of rapamycin.

[0061] In yet another embodiment, the solution or spray formulations of the invention comprise a polymer solution of drug that is sprayed onto intact skin, resulting in a sustained release of rapamycin from the polymer matrix.

[0062] In yet other embodiments, cream or lotion formulations of the present invention contain polyoxyethylene, glycerol, paraffin, propylene glycol, and glycerol.

[0063] In yet another embodiment, an ointment formulation of the present invention contains petroleum, paraffin, steryl alcohol, and cholesterol.

[0064] Controlled Release Formulations and Drug Delivery Systems The controlled release or sustained release formulation of the pharmaceutical composition of the present invention can be manufactured using conventional technology.In some cases, the dosage form used is provided as a slow or controlled release of one or more active ingredients in the dosage form, and can use, for example, hydroxypropylmethylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multi-layer coatings, microparticles, liposomes, microspheres, or combinations thereof to provide desired release characteristics at various rates.Suitable controlled release formulations known to those skilled in the art can be easily selected for use with the pharmaceutical composition of the present invention, including those described herein.Therefore, single unit dosage forms suitable for topical administration, such as liniments, lotions, oil-in-water or water-in-oil emulsions suitable for controlled release, such as creams, ointments or pastes, transdermal patches, and solutions or suspensions, are encompassed by the present invention.

[0065] Most controlled-release pharmaceutical products share a common goal of improving drug therapy beyond that achieved by their non-controlled counterparts. Ideally, the medical use of an optimally designed controlled-release formulation is characterized by curing or suppressing symptoms in a minimum amount of time using a minimum amount of drug substance. Advantages of controlled-release formulations include extended activity of the drug, reduced dosing frequency, and improved patient compliance. In addition, controlled-release formulations can be used to affect the onset of action or other characteristics, such as the blood concentration of the drug, and therefore can affect the occurrence of side effects.

[0066] Most controlled release formulations are designed to initially release an amount of drug that rapidly produces the desired therapeutic effect, and then gradually and continuously release other amounts of drug to maintain this level of therapeutic effect over an extended period of time. In order to maintain this constant level of drug in the body, the drug must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body.

[0067] The controlled release of an active ingredient can be stimulated by various inducers, such as pH, temperature, enzymes, water, or other physiological conditions or compounds. In the context of the present invention, the term "controlled release component" is defined herein as one or more compounds that facilitate the controlled release of an active ingredient, including, but not limited to, polymers, polymer matrices, gels, permeable membranes, liposomes, microspheres, or combinations thereof.

[0068] In certain embodiments, the formulations of the present invention can be, but are not limited to, short-acting, rapid-offset, and controlled, including sustained-release, delayed-release, and pulsatile-release formulations.

[0069] The term sustained release is used in its conventional sense to refer to a formulation that can gradually release drug over an extended period of time to provide, but not necessarily, a substantially constant blood concentration of drug over an extended period of time, which can be as long as a month or longer, providing a longer release than would occur if the same amount of drug were administered in bolus form.

[0070] For sustained release, the compounds may be formulated with a suitable polymer or hydrophobic material that imparts sustained release properties to the compounds.

[0071] In certain aspects of the invention, the compositions of the invention are administered to a patient, alone or in combination with another agent, using a sustained release formulation.

[0072] The term delayed release, as used herein, refers in its conventional sense to a formulation that provides for an initial release of drug with some delay after drug administration, which may, but does not necessarily, include a delay of from about 10 minutes to about 12 hours.

[0073] The term pulsatile release, as used herein, is given its conventional meaning to refer to a formulation that provides release of drug in such a manner as to result in a pulsatile plasma profile of the drug following drug administration.

[0074] The term immediate release, as used herein, refers in its conventional sense to a formulation that provides release of drug immediately following drug administration.

[0075] As used herein, acute refers to any period up to and including about 24 hours, about 48 hours, about 72 hours after administration of the drug, and any whole or partial increments therein.

[0076] Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents are considered to be within the scope of the present invention and encompassed by the appended claims. For example, it should be understood that it is within the scope of this application to vary reaction conditions, including but not limited to reaction times, reaction sizes / volumes, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions (e.g., nitrogen atmosphere), and reducing / oxidizing agents, using art-recognized substitutes and no more than routine experimentation.

[0077] Whenever values ​​and ranges are provided herein, it is to be understood that all values ​​and ranges subsumed within those values ​​and ranges are included within the scope of the invention. Moreover, all values ​​that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the application.

[0078] The following examples further illustrate aspects of the present invention, but do not in any way limit the teachings or disclosure of the invention set forth herein. EXAMPLES

[0079] The present invention will now be described with reference to the following examples, which are provided for illustrative purposes only and the invention is not limited to these examples, but rather includes all variations that become apparent as a result of the teachings provided herein.

[0080] material and method Unless otherwise stated, all cell lines, starting materials, reagents and cell lines were obtained from commercial suppliers and used without further manipulation.

[0081] Patients were provided with a proprietary topical rapamycin formulation (0.001%; ​​equivalent to 10 μM) or an identical placebo formulation, except that it did not contain rapamycin. For seborrheic keratosis, patients were instructed to apply the formulation to a localized area of ​​skin up to 1 cm around the lesion before going to bed. Patients were instructed to apply the cream to the dorsum of the hand once daily before going to bed.

[0082] The cream contained the following ingredients: polyoxyethylene (40) stearate, polyoxyethylene (20) sorbitan monooleate, glycerol monostearate, paraffin, cetyl alcohol, mineral oil, water, propylene glycol, sorbitol, glycerol, and methylparaben.

[0083] Patients were provided with a 4-week supply of the cream, labeled with a number without any indication of whether it contained rapamycin, and were instructed to contact the study personnel at any time if they experienced any irritation or negative reactions.

[0084] Example 1: Human skin was treated with a formulation containing 1 micromolar rapamycin or the same formulation containing vehicle control for 240 days. 0.5 cc of the formulation was applied daily. Skin biopsies (N=8) were processed for immunohistochemistry and nuclear p16 was quantified using a Leica Aperio software system. As can be seen in Figure 1, rapamycin treatment reduced the levels of p16 in human skin.

[0085] Example 2: Hematoxylin-eosin staining of human skin biopsies taken from the dorsum of a patient's hand following application of a topical cream containing rapamycin (10 μM) or placebo is shown in Figure 2. Histological evidence of actinic (sun-induced) elastosis is indicated by the arrow. Similar to those shown in Figure 2, a decreased presence of these histological markers of aged / damaged skin was noted in multiple patient biopsies.

[0086] Example 3: Images of human skin biopsies taken from the dorsal surface of a patient's hand after application of a topical cream containing rapamycin (10 μM) or placebo are shown at 40x magnification in Figure 3. The rapamycin-treated biopsy shows a more orderly arrangement of the epithelial layers, including apical placement of nuclei within the basal layer and a distinct keratin layer.

[0087] Example 4: Images at 40x magnification of human skin biopsies taken from the dorsum of a patient's hand after application of a topical cream containing rapamycin (10 μM) or placebo are shown in Figure 4. Untreated skin shows signs of UV damage in the form of purple-stained elastin fibers and disorganized collagen fibers, known as solar elastosis. Biopsies treated with rapamycin show a more ordered arrangement of collagen fibers and less solar elastosis.

[0088] Example 5: Human skin biopsies taken from the dorsum of the patient's hand after application of a topical cream containing rapamycin (10 μM) or placebo were stained with an antibody that recognizes cytokeratin 5 / 6, a marker of epidermal basal cells (brown staining). Note that staining in rapamycin-treated skin is more focally located in the basal layer of the skin, whereas placebo-treated skin shows cytokeratin 5 / 6 (CK 5 / 6) staining in the granular layer, suggesting incomplete differentiation typical of aged skin.

[0089] Example 6: In placebo-treated skin, CK 5 / 6 staining at high magnification (40x) showed positive staining in the granular layer and extended into the stratum corneum, as shown in Figure 6, whereas in rapamycin-treated biopsies, the pattern of CK 5 / 6 staining remained restricted to the basal layer.

[0090] Example 7: A 67-year-old woman showed clinical improvement after six months of application of 10 μM (0.001%) rapamycin cream, as shown in Figure 7. For comparison, the placebo-treated left hand is also shown.

[0091] Example 8: A Caucasian female in her mid-60s was treated with rapamycin (0.001%) for 4.5 months. The patient was a cancer survivor who had undergone multiple chemotherapy treatments and had recurrent purpura with subsequent scarring. The results of treatment are shown in Figure 8, which shows that after the 4.5 month treatment period, there was a significant reduction in purpura. Treatment was discontinued at the end of the study.

[0092] Example 9: Figures 9A-9C show that rapamycin protects human cells from UV damage. Cells grown in the presence or absence of 1 nM rapamycin were exposed to increasing amounts of UV light and cell counts were measured after 24 hours. Percent cell loss is shown in Figure 9A. Cultures treated with rapamycin showed reduced cell death after UV irradiation (P=0.0044). Figure 9B shows a representative photograph of a control culture 24 hours after UV irradiation. Note the typical apoptotic features, such as rounded floating cells and typical cell blebbing. Figure 9C shows a parallel culture grown in 1 nM rapamycin, which shows a significant loss of apoptotic cells.

[0093] List of embodiments Exemplary embodiments are provided below, the numbering of which should not be construed as designating a level of importance.

[0094] Aspect 1 provides: A method for treating and / or preventing a dermal disorder in a mammalian subject in need thereof, comprising the step of topically administering to the subject a therapeutically effective amount of a composition comprising rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytide, UV damage, prominent hand veins, epithelial mass carcinogenesis such as in squamous cell carcinoma, keloids, and imbalance in the ratio of collagen isoforms.

[0095] Aspect 2 provides: The method of embodiment 1, wherein the epithelial wide-spectrum carcinogenesis comprises epidermal dysplasia and actin damage.

[0096] Aspect 3 provides: The method of embodiment 1-2, wherein the subject is a human.

[0097] Aspect 4 provides: The method of any one of embodiments 1 to 3, wherein the composition comprises from about 0.1% to about 0.0001% by weight of rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof.

[0098] Aspect 5 provides the following: The method of any one of embodiments 1 to 4, wherein the composition comprises from about 0.001% to about 0.0001% by weight of rapamycin, or a salt, solvate, enantiomer, or diastereomer thereof.

[0099] Aspect 6 provides the following: The method of any one of embodiments 1 to 5, wherein the composition further comprises a dermatologically acceptable carrier.

[0100] Aspect 7 provides the following: The method of any one of the preceding claims, wherein the dermatologically acceptable carrier is at least one selected from the group consisting of solvents, lubricants, emollients, emulsifiers, moisturizers, thickening waxes, emollients, fragrances, preservatives, and artificial colorants.

[0101] Aspect 8 provides the following: The method of any one of embodiments 1 to 7, wherein the dermatologically acceptable carrier comprises petrolatum.

[0102] Aspect 9 provides the following: The method of any one of embodiments 1 to 8, wherein the composition is applied to the site of a dermal disorder in a subject.

[0103] Aspect 10 provides the following: A kit for treating and / or preventing a dermal disorder in a mammalian subject in need thereof, comprising a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, an applicator, and instructions for topically administering the composition to the subject, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytides, prominent hand veins, epithelial mass carcinogenesis, keloids, and imbalance in the ratio of collagen isoforms.

[0104] Other embodiments The recitation of a list of elements in a definition of a variable herein includes definition of that variable as any single element or combination (or subcombination) of the listed elements. The recitation of an embodiment herein includes that embodiment as a single embodiment or in combination with other embodiments or portions thereof.

[0105] The disclosures of any and all patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety. Although the present invention has been disclosed in connection with certain embodiments, it is apparent that those skilled in the art can devise other embodiments and variations of the present invention without departing from the true spirit and scope of the present invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.

Claims

1. 1. A method for treating a dermal disorder in a mammalian subject in need thereof, comprising the step of topically administering to the subject a therapeutically effective amount of a composition comprising rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytide, UV damage, prominent hand veins, epithelial mass carcinogenesis such as in squamous cell carcinoma, keloids, and imbalance in the ratio of collagen isoforms.

2. The method of claim 1, wherein epithelial widespread carcinogenesis includes epidermal dysplasia and actin damage.

3. The method of claim 1 , wherein the subject is a human.

4. The method of claim 1, wherein the composition comprises from about 0.1% to about 0.0001% by weight of rapamycin.

5. The method of claim 1, wherein the composition comprises from about 0.001% to about 0.0001% by weight of rapamycin.

6. The method of claim 1 , wherein the composition further comprises a dermatologically acceptable carrier.

7. 7. The method of claim 6, wherein the dermatologically acceptable carrier is at least one selected from the group consisting of solvents, lubricants, emollients, emulsifiers, moisturizers, thickening waxes, emollients, fragrances, preservatives, and artificial colors.

8. 7. The method of claim 6, wherein the dermatologically acceptable carrier comprises petrolatum.

9. The method of claim 1 , wherein the composition is applied to the subject at the site of a dermal disorder.

10. 1. A kit for treating and / or preventing a dermal disorder in a mammalian subject in need thereof, comprising a composition comprising a therapeutically effective amount of rapamycin, or a salt, solvate, enantiomer or diastereomer thereof, an applicator, and instructions for topically administering the composition to the subject, wherein the dermal disorder is at least one selected from the group consisting of static wrinkles, fine lines, loss of skin turgor, freckles, melasma, senile purpura, static rhytides, prominent hand veins, epithelial mass carcinogenesis, keloids, and imbalance in collagen isoform ratios.

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