Method of reducing pigmentation and combinations thereof ii

Abstract

The invention relates to methods of skincare. In particular, the present specification teaches a method of reducing skin pigmentation in a subject. Combinations for reducing skin pigmentation are also defined herein, and the combinations comprises 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

Description

[0001] Method of Reducing Pigmentation and Combinations Thereof II

[0002] Field of Invention

[0003] The invention relates to methods of skincare. In particular, the present specification teaches a method of reducing skin pigmentation in a subject. Combinations for reducing skin pigmentation are also defined herein.

[0004] Background

[0005] Skin pigmentation is a physiological process for producing and distributing melanin, in which the melanin protects the skin from potential damage caused by ultraviolet (UV) radiation. UV radiation exposure triggers the release of mediators, such as reactive oxygen species (ROS) and alpha-melanocyte stimulating hormones (a-MSH) from the keratinocytes and melanocytes present in the skin. This, in turn, stimulates the production of melanin (melanogenesis) in the melanocytes. These melanin pigments are packed into melanosome granules, which are transferred out to the surrounding keratinocytes, forming protective nuclei caps against further UV-induced damage.

[0006] The two main types of melanin that contribute to skin and hair pigmentation are eumelanin and pheomelanin. The balance between these two types of melanin, where eumelanin contributes to the dark brown-black pigmentation, while pheomelanin contributes to the reddish-yellow tinge, determines skin color and UV protection level for each individual. These processes can be pathologically altered by certain triggers, including chronic UV exposure and aging (e.g., solar lentigines), hormonal changes (e g., melasma), as well as immune- and oxidative stress-induced loss of melanocytes, resulting in hypopigmentary conditions, e.g., vitiligo. These conditions result in skin areas with highly visible depigmentation or hyperpigmentation in all regions of the body, including the faces, arms, and limbs. Hence, these conditions come with very significant psychosocial and emotional impacts as well as a reduction in the quality of life of the subject.

[0007] Traditional treatments for hyperpigmentary disorders include pharmacological agents,including corticosteroids and hydroquinone. These lighten skin tone through the inhibition of melanocyte maturation or through inhibition of tyrosinase activity or the cAMP pathway which are the main factors driving pigmentation. Many currently used agents have limited efficacy in the cutaneous treatment of the disorders due to issues including difficulty penetrating the human skin barrier. In addition, many actives exhibit adverse side effects including prickling sensation, contact dermatitis, irritation, high toxicity and sensitivity.

[0008] It would be desirable to overcome or ameliorate at least one of the above-described problems.

[0009] Summary

[0010] Disclosed herein is a method of reducing skin pigmentation in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0011] Disclosed herein is a method of inhibiting melanin production in a melanocyte or transfer of melanin in the melanocyte to a keratinocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to inhibit melanin production in the melanocyte or transfer to the keratinocyte.

[0012] Disclosed herein is a method of inhibiting tyrosinase in a melanocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to inhibit tyrosinase in the melanocyte.Disclosed herein is a method of lightening or whitening skin in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0013] Disclosed herein is a method of treating a hyperpigmentation or a hypermelanosis disorder in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0014] Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, for use in reducing pigmentation in a subject.

[0015] Disclosed herein is the use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, in the manufacture of a medicament for reducing pigmentation in a subject.

[0016] Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, for use in treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0017] Disclosed herein is the use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extractcomprising ginsenoside RO in the manufacture of a medicament for treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0018] Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

[0019] Disclosed herein is a composition comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

[0020] Brief Description of Drawings

[0021] Embodiments of the present invention are hereafter described, by way of non-limiting example only, with reference to the accompanying drawings in which:

[0022] Figure 1: Synergistic effect of the combination of sclareolide (SC) and ginsenoside RO (GIN RO) on the 3D Reconstructed Human Pigmented Epidermal (RHPE) model under baseline NO UVB-irradiated condition.

[0023] Detailed Description

[0024] The present specification teaches a method of reducing skin pigmentation in a subject.

[0025] The specification teaches a method of reducing skin pigmentation in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside, to the subject. The ginsenoside may be ginsenoside RO, ginsenoside Rbl, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rgl and / or ginsenoside Rhl.Disclosed herein is a method of reducing skin pigmentation in a subject, the method comprising administering a combination comprising 1) sclareolide, a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0026] Without being bound by theory, the inventors have found that sclareolide and ginsenoside RO can reduce epidermal pigmentation in an in vitro primary melanocyte assay (by reducing tyrosinase activity and melanin levels) and reducing pigmentation in the 2D in vitro co-culture system with primary melanocytes and primary keratinocytes (by reducing melanin transfer or melanin synthesis). Here the synergistic effect of sclareolide and ginsenoside RO is shown in the 3D Reconstructed Human Pigmented Epidermal (RHPE) model, where stronger anti -pigmentation effects are observed when compared to single actives alone. Sclareolide and ginsenoside RO, in combination, may be used in commercial cosmetic skincare products to promote skin lightening or whitening in normal or hyperpigmented skin.

[0027] While drug combinations are theoretically feasible, on a practical level, the effects of drug combinations are unpredictable. The effect of co-administrating multiple drugs may often be described as synergistic, additive or antagonistic. This illustrates that not all drug combinations provide useful or beneficial effects. It is not predictable from the outset that specific combinations will provide a synergistic effect. In some cases, the combination may even amplify the adverse side effects, which is not desirable. In one embodiment, the combination of sclareolide and ginsenoside RO act to synergistically reduce skin pigmentation in a subject with minimal adverse side effects. Sclareolide and ginsenoside RO have different structures and chemical classes (such that sclareolide is a diterpene lactone while ginsenoside RO is a triterpene saponin), and are commonly used in different applications, hence, the effects of combination may be unpredictable.Sclareolide is a bicyclic diterpene lactone. Sclareolide has a chemical structure of

[0028]

[0029] Sclareolide (CAS Number 564-20-5) is a compound prepared by chemical modification or by biotransformation of the labdan type diterpene sclareol. Sclareol is present in stems, leaves and flowering parts of clary sage (Salvia sclarea L.) and its isolation from this source has been described (US 3,060,172). The source of sclareolide can be derived (extracted) naturally from either species of the Salvia genus or can be synthetically obtained as substantially pure sclareolide. The substantially pure sclareolide may contain more than 70 percent sclareolide. The sclareolide may be obtained from commercially available sources, such as SymBright® 2036 manufactured by Symrise, or Sclarance® manufactured by Ashland.

[0030] By way of example, sclareolide may be found in various plant, such as plants of the Salvia sp. (including S. sclarea, S. yosgadensis, S. amplexicaulis, S. candidissima, and . amplexicaulis). It may be derived from natural sources, such as plant-derived sclareol, or may be completely or partially synthesized from a related compound using methods of synthesis known in the art. The compound may be used in cosmetics, perfumes, skin care products, and other personal care formulations.

[0031] Ginsenosides are a class of steroidal glycosides and triterpene saponins found predominantly in plants of the Panax genus, including Panax ginseng, Panax quinquefolius, and Panax notoginseng. These compounds represent the principal bioactive constituents of ginseng Structurally, each ginsenoside comprises a hydrophobic aglycone (sapogenin) linked to one or more hydrophilic sugar moieties, with the nature, number, and position of the sugar groups critically influencing biological function.

[0032] Ginsenosides are generally classified according to the structure of their aglycone skeleton into two major categories: dammarane-type and oleanane-type saponins.Dammarane-type ginsenosides, which constitute the majority of known ginsenosides, include the protopanaxadiol (PPD) subclass, characterized by sugar substitution typically at carbon positions C-3 and / or C-20, and the protopanaxatriol (PPT) subclass, featuring sugar moieties primarily at C-6 and / or C-20 Representative PPD-type ginsenosides include Rbl, Rb2, Rc, Rd, and Rg3, whereas representative PPT-type ginsenosides include Re, Rgl, and Rg2. In contrast, oleanane-type ginsenosides, such as ginsenoside RO, possess a pentacyclic triterpene aglycone and are known for specific activities including anti-inflammatory and immunomodulating effects.

[0033] In some embodiments, the ginsenoside is a ginsenoside RO. In another embodiment, the

[0034] ginsenoside has a chemical structure

[0035]

[0036] The present disclosure pertains to compounds and their various forms, including ionic forms, tautomers, isomers, polymorphs, pseudopolymorphs, amorphous forms, solvates, co-crystals, chelates, esters, prodrugs, and protected forms. The disclosure also encompasses methods of utilizing these compounds for various purposes. It should be noted that terms like "crystalline form," "polymorph," can be used interchangeably to include all crystalline and amorphous forms, such as polymorphs, pseudopolymorphs, solvates (including hydrates), co-crystals, unsolvated polymorphs (including anhydrates), conformational polymorphs, amorphous forms, and mixtures thereof, unless a specific crystalline or amorphous form is specified. In certain embodiments, the compounds and their subgroups include polymorphs, solvates, co-crystals, isomers, tautomers, and / or oxides. In other embodiments, they may include polymorphs, solvates, and / or co-crystals.

[0037] As used herein, the term “salt” refers to pharmaceutically acceptable salts commonly used to form alkali metal salts of free acids and to form addition salts of free bases. The nature of the salt is not critical, provided that it is pharmaceutically acceptable. The term “salts” also includes solvates of addition salts, such as hydrates, as well as polymorphsof addition salts. Suitable pharmaceutically acceptable acid addition salts can be prepared from an inorganic acid or from an organic acid. Non-limiting examples of such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric, and phosphoric acid Appropriate organic acids can be selected from aliphatic, cycloaliphatic, aromatic, arylaliphatic, and heterocyclyl containing carboxylic acids and sulfonic acids, for example formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic, stearic, salicylic, p-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, toluenesulfonic, 2-hydroxyethanesulfonic, sulfanilic, cyclohexylaminosulfonic, algenic, 3 -hydroxybutyric, galactaric and galacturonic acid.

[0038] As used herein, the term “solvate” refers to a form of a compound in which molecules of a solvent are physically or chemically associated with molecules or ions of the compound. The association may occur through hydrogen bonding, coordination, van der Waals interactions, or other intennolecular forces, and may involve stoichiometric or non-stoichiometric amounts of solvent. A solvate may include but not limited to crystalline or amorphous forms in which solvent molecules are incorporated into the solid-state lattice or are otherwise retained within the material. When the associated solvent is water, the solvate is referred to as a hydrate.

[0039] As used herein, the term “isomer” includes especially optical isomers (for example essentially pure enantiomers, essentially pure diastereomers, and mixtures thereof) as well as conformation isomers (i.e. isomers that differ only in their angles of at least one chemical bond), position isomers (particularly tautomers), and geometric isomers (e.g. cis-trans isomers).

[0040] Compounds described herein can comprise one or more asymmetric centers, and thus can exist in various isomeric forms, c. ., enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of an individual enantiomer, diastereomer or geometric isomer, or can be in the form of a mixture of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomer. Isomers can be isolated from mixtures by methods known to those skilledin the art, including chiral high pressure liquid chromatography (HPLC) and the fonnation and crystallization of chiral salts; or preferred isomers can be prepared by asymmetric syntheses. The invention additionally encompasses compounds described herein as individual isomers substantially free of other isomers, and alternatively, as mixtures of various isomers. "Optically-enriched," as used herein, means that the compound is made up of a significantly greater proportion of one enantiomer. In certain embodiments, the compound of the present invention is made up of at least about 90% by weight of a preferred enantiomer. In other embodiments, the compound is made up of at least about 95%, 98%, or 99% by weight of a preferred enantiomer. Preferred enantiomers may be isolated from racemic mixtures by any method known to those skilled in the art, including chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of chiral salts or prepared by asymmetric syntheses. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions p. 268 (E.L Eliel, Ed., Univ of Notre Dame Press, Notre Dame, IN 1972).

[0041] As used herein, the term “derivative” refers to a compound that is formed from another compound by a chemical modification, where part of the original molecule is replaced, removed, or altered. The new compound retains the core structure or backbone of the original substance but differs by one or more functional groups or substituents.

[0042] It will be appreciated that a prodrug of the compound is also within the scope and spirit of the invention. Thus, the compound can be administered to a subject in the form of a pharmaceutically acceptable prodrug. The term “prodrug” is used in its broadest sense and encompasses those derivatives that are converted in vivo to the compound of the invention. Such derivatives would readily occur to those skilled in the art. Other texts which generally describe prodrugs (and the preparation thereof) include: Design of Prodrugs, 1985, H. Bundgaard (Elsevier); The Practice of Medicinal Chemistry, 1996, Camille G. Wermuth et al., Chapter 31 (Academic Press), and A Textbook of Drug Design and Development, 1991, Bundgaard et al., Chapter 5, (Harwood Academic Publishers). For example, the carboxylic acid moiety may be protected as an ester.In one embodiment, the sclareolide, or a salt, solvate, derivative or prodrug thereof, or botanical extract comprising sclareolide, is provided at a concentration of less than 1 mM, less than 500 pM, less than 100 pM, less than 50 pM, less than 10 pM, less than 5 pM, less than 1 pM, less than 500 nM, less than 300 nM, less than 100 nM, such as between 50 nM to ImM, between 50 nM to 500 pM, between 50 nM and 100 pM, between 50 nM and 50 pM, between 50 nM and 10 pM, between 50 nM and 5 pM, between 50 nM and 1 pM, between 50 nM and 500 nM, between 50 nM and 300 nM, or between 50 nM and 100 nM.

[0043] In one embodiment, the ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or botanical extract comprising ginsenoside R0, is provided at a concentration of less than 1 mM, less than 500 pM, less than 100 pM, less than 50 pM, less than 10 pM, less than 5 pM, less than 1 pM, less than 500 nM, less than 300 nM, less than 100 nM, such as such as between 50 nM to ImM, between 50 nM to 500 pM, between 50 nM and 100 pM, between 50 nM and 50 pM, between 100 nM and 50 pM, between 500 nM and 50 pM, between 1 pM and 50 pM, between 1 pM and 30 pM, or between 1 pM and 10 pM.

[0044] The sclareolide, or a salt, solvate, derivative or prodrug thereof may, for example, be sclareol, sclareolide acetate, sclareolide epoxide, sclareolide oxime, sclareolide semicarbazone, sclareolide phenylhydrazone, sclareolide tosylate, sclareolide benzoate, sclareolide-indole conjugates, aminoalkyl sclareolide, ambradiol, ambroxide, or a salt, solvate, derivative or prodrug thereof.

[0045] The botanical extract comprising ginsenoside R0 may comprise one or more other ginsenoside such as ginsenoside Rbl, Rc, Rd, Re, Rgl and / or Rhl. The botanical extract may be Ginseng Herbasec® or other ginseng extract.

[0046] In some embodiments, the sclareolide is a stereoisomer thereof, such as isosclareolide.

[0047] In some embodiments, the ginsenoside R0 is a stereoisomer thereof, such as 20(S)-ginsenoside R0 and 20(R)-ginsenoside R0.In one embodiment, the method is a non-therapeutic or cosmetic method.

[0048] In some embodiments, the sclareolide and / or ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside RO, is / are capable of reducing skin pigmentation in a subject.

[0049] The term “reducing skin pigmentation” as used herein, refers to decreasing the amount, concentration, or visible appearance of melanin in the skin, including epidermal or dermal layers, resulting in a measurable or visible lightening of the skin, such as in a hyperpigmented skin area, or a more even skin tone.

[0050] In some embodiments, the sclareolide and / or ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside RO is capable of reducing skin pigmentation in skin or in a 3D RHPE model by at least about 0.1% (including 0.1%, 0.5%, 1%, 2%, 3%, 5%, 7.5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, and 100%) after at least 7 days (including after 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, or more) of applying the sclareolide and / or ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside R0.

[0051] In some embodiments, the sclareolide and / or ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside R0, is / are capable of inhibiting melanin production or transfer in a melanocyte or keratinocyte.

[0052] In some embodiments, the sclareolide and / or ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside R0, is / are capable of inhibiting tyrosinase in a melanocyte.In some embodiments, the sclareolide and / or ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside RO, is / are capable of lightening or whitening skin in a subject.

[0053] In some embodiments, the sclareolide and / or ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside RO, is / are capable of treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0054] The term “subject” as used throughout the specification is to be understood to mean a human or may be a domestic or companion animal. While it is particularly contemplated that the methods of the invention are for treatment of humans, they are also applicable to veterinary treatments, including treatment of companion animals such as dogs and cats, and domestic animals such as horses, cattle and sheep, or zoo animals such as primates, felids, canids, bovids, and ungulates. The “subject” may include a person, a patient or individual, and may be of any age or gender.

[0055] In some embodiments, the subject is a mammalian subject. In another embodiment, the subject is a human subject.

[0056] In some embodiments, the combination lightens the subject’s pigmented skin.

[0057] As used herein, the term “sample” includes tissues, cells, body fluids and isolates thereof etc., isolated from a subject, as well as tissues, cells and fluids etc present within a subject (i.e. the sample is in vivo). Examples of samples include: whole blood, blood fluids (e g. serum and plasm), lymph and cystic fluids, sputum, stool, tears, mucus, hair, skin, ascitic fluid, cystic fluid, urine, nipple exudates, nipple aspirates, sections of tissues such as biopsy and autopsy samples, frozen sections taken for histologic purposes, archival samples, explants and primary and / or transformed cell cultures derived from patient tissues etc.

[0058] A sample can be a biological sample which refers to the fact that it is derived or obtained from a living organism. The organism can be in vivo (e g. a whole organism) or can bein vitro (e.g., cells or organs grown in culture). A "biological sample" also refers to a cell or population of cells or a quantity of tissue or fluid from a subject. Most often, a sample has been removed from a subject, but the term "biological sample" can also refer to cells or tissue analyzed in vivo, i.e., without removal from the subject. Often, a “biological sample” will contain cells from a subject, but the term can also refer to non-cellular biological material, such as non-cellular fractions of blood, saliva, or urine.

[0059] Biological samples also include explants and primary and / or transformed cell cultures derived from patient tissues. A biological sample can be provided by removing a sample of cells from subject, but can also be accomplished by using previously isolated cells or cellular extracts (e g. isolated by another person, at another time, and / or for another purpose). Archival tissues, such as those having treatment or outcome history may also be used. Biological samples include, but are not limited to, tissue biopsies, scrapes (e g. buccal scrapes), skin, whole blood, plasma, serum, urine, saliva, cell culture, or cerebrospinal fluid.

[0060] In some embodiments, the skin cell or tissue may be collected from a subject and the melanin level in the collected sample may be analysed using methods known in the art. The level of production or synthesis of melanin may be detected or measured using methods known in the art.

[0061] The methods collecting skin samples for measuring the level of melanin may include skin biopsy (such as punch, shave, or excision), surgical skin resection, tape stripping method, suction blister, swabbing or lavage, primary skin cell isolation, and skin explants.

[0062] In some embodiments, the melanin level may directly be measured from the face of the subject.

[0063] The terms “detecting”, “determining”, “measuring”, “evaluating”, “assessing” and “assaying” are used interchangeably herein to refer to any form of measurement, and include determining if an element is present or not. These terms include both quantitative and / or qualitative determinations. Assessing may be relative or absolute.The method as defined herein may comprise measuring or visualising the levels of melanin in a sample.

[0064] The methods of measuring a whitening or brightening or lightening effect on the skin of the subject, or the level of melanin in the skin sample, includes but not limited to objective colorimetric measurement, melanin index (MI) measurement, skin colorimeters, individual typology angle (ITA), multispectral or hyperspectral imaging, melanometry devices, reflectance spectrophotometry, combined melanin and hemoglobin measurement, clinical UV-induced melanization models, photography with standardized lighting, spectrophotometry or absorption spectroscopy, fluorescence spectroscopy, high-performance liquid chromatography (HPLC), electron paramagnetic resonance (EPR) spectroscopy, urine melanin test, and genetic testing. In some embodiments, the level of melanin or the melanin content in the skin sample is measured using Solvable™ (PerkinElmer) melanin assay.

[0065] In some embodiments, the method to assess appearance or lightening or whitening of the subject’s skin may comprise a full-face image analysis using a dedicated face analyser device (such as a facial image analysis technology employed in commercial systems) or a smartphone equipped with face analysis software application. The full face image analysis may comprise capturing multiple images of the face of the subject under multi -spectral lighting conditions (such as natural lighting, cross-polarised light, ultraviolet light), 3D image analysis, image processing, identification of wrinkles, fine lines, dark spots, textures, firmness, visible pores, redness, eye puffmess, dark circles, crow’ s feet, UV damage and clogged pores. The analysis may further comprise captured images processed by a computer algorithm or artificial intelligence software by comparing the obtained images to a database of images to obtain a score of dark spots, skin age, photo damage, or other suitable parameters. The analysis method may provide a quantitative and reproducible measurement that can be used to evaluate the efficacy of the compositions and methods disclosed herein.

[0066] The decrease in the level of melanin may be, for example, a decrease of at least about 0.1%, at least about 0.5%, at least about 1%, at least about 2%, at least about 3%, at least about 5%, at least about 7.5%, at least about 10%, at least about 15%, at least about20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90% or up to and including a 100% decrease or any decrease between 0.1-100% as compared to a reference level.

[0067] In some embodiments, the decrease in the melanin level is about 5% to about 10%, about 5% to about 20%, about 5% to about 30%, about 5% to about 50%, about 10% to about 20%, about 10% to about 30%, or about 10% to about 25%.

[0068] The term “reference” as used herein, refers to a subject that is not administered with a composition comprising sclareolide and / or ginsenoside R0, or a salt, solvate, derivate or prodrug thereof, for reducing skin pigmentation or reducing melanin level in the skin or before a subject is administered with a composition comprising sclareolide and / or ginsenoside R0, or a salt, solvate, derivate or prodrug thereof, for reducing skin pigmentation or reducing melanin level in the skin.

[0069] Tn some embodiments, a subject may be a healthy individual with normal skin pigmentation. The subject may wish to reduce skin pigmentation by administering a composition as disclosed herein. In some embodiments, the normal pigmentation may be on the skin of the face, neck, chest, tummy, lower back, upper back, buttocks, thighs, or the skin on any part of the body of an individual.

[0070] In some embodiments, a subject is suffering from a hyperpigmentation or a hypermelanosis disorder. The hyperpigmentation may appear as brown, black, gray, red or pink spots or patches on the skin of the subject. In some embodiments, the hyperpigmentation or hypermelanosis may be on the skin of the face, neck, chest, tummy, lower back, upper back, buttocks, thighs, or the skin on any part of the body of an individual. In some embodiments, the spots associated with hyperpigmentation or hypermelanosis are known as age spots, sun spots, solar lentigines, or liver spots.The hyperpigmentation or a hypermelanosis disorder may include melasma, post-inflammatory hyperpigmentation, sun spots, pregnancy or aging associated pigmentation, yeast or bacterial associated pigmentation.

[0071] In some embodiments, the hyperpigmentation or hypermelanosis disorder may be caused by melasma, inflammation (such as acne or eczema), genetics associated cause, adrenal disorders (such as Addison’s disease), medications (such as oral contraceptives or drugs that cause sensitivity to light), sun damage, or thyroid disorders.

[0072] As used herein, the term “treating” refers to partially or completely alleviating, ameliorating, improving, relieving, delaying onset of, inhibiting progression of, reducing severity of, and / or reducing incidence of one or more symptoms or features of a particular infection, disease, disorder, and / or condition.

[0073] The term “administering” refers to contacting, applying, or providing 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and / or 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0074] In one embodiment, the method comprises topically or transdermally administering 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and / or 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0075] In some embodiments, the method comprises topically or transdermally administering a combination or a composition to the subject. Modes of administration including topical and transdermal administration may be possible. Non-limiting examples of transdermal delivery mechanisms include microneedles or dissolvable microneedles, and the microneedles may be fabricated into a form of microneedle patch.

[0076] In a suitable embodiment, the administration method may be combined with a device which enhances transdermal penetration of a composition The composition may thus be applied to the area requiring depigmentation of the skin. Merely by way of example such devices may include devices which produce electrical current such as galvanicdevices, lasers, devices which produce vibrations such as ultrasound, and massage devices which enhance blood circulation around the target cells or target areas.

[0077] The sclareolide and / or ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide and / or ginsenoside RO as disclosed herein may be administered in a variety of routes including oral, transdermal, topical, parenteral (including subcutaneous, intravenous, intramuscular and intraperitoneal), vaginal, rectal, oropharyngeal, intrapulmonary and intranasal routes.

[0078] By “effective amount”, in the context of treating or preventing a condition is meant the administration of an amount of 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and / or 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to an individual in need of such treatment or prophylaxis, either in a single dose or as part of a series, that is effective for the prevention of incurring a symptom, holding in check such symptoms, and / or treating existing symptoms, of that condition. The effective amount will vary depending upon the health and physical condition of the individual to be treated, the taxonomic group of individual to be treated, the formulation of the composition, the assessment of the medical situation, and other relevant factors. It is expected that the amount will fall in a relatively broad range that can be determined through routine trials.

[0079] Dosing may occur at intervals of minutes, hours, days, weeks, months or years or continuously over any one of these periods. Suitable dosages may lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. In one embodiment, the dosage may be in the range of 1 mg to 500 mg per kg of body weight per dosage. In another embodiment, the dosage may be in the range of 1 mg to 250 mg per kg of body weight per dosage. In yet another embodiment, the dosage may be in the range of 1 mg to 100 mg per kg of body weight per dosage, such as up to 50 mg per body weight per dosage.In some embodiments, the sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, is provided at a concentration of about 0.001 % (w / v) to about 99 % (w / v) relative to the combination. In other embodiments, the range is about 0.01 % (w / v) to about 99% (w / v). In other embodiments, the range is about 0.1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 5 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 10 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 20 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 30 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 40 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 50 % (w / v) to about 99 % (w / v). Tn other embodiments, the range is about 60 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 70 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 80 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 90 % (w / v) to about 99 % (w / v).

[0080] In one embodiment, the sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, is provided at a concentration of less than about 2 % (w / v) relative to the combination, such as less than about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v), or about 0.01% (w / v).

[0081] In some embodiments, the ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, is provided at a concentration of about 0.001 % (w / v) to about 99 % (w / v) relative to the combination. In other embodiments, the range is about 0.01 % (w / v) to about 99% (w / v). In other embodiments, the range is about 0.1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 5 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 10 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 20 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 30 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 40 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 50 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 60 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 70 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 80 %(w / v) to about 99 % (w / v). In other embodiments, the range is about 90 % (w / v) to about 99 % (w / v).

[0082] In one embodiment, the ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, is provided at a concentration of less than about 2 % (w / v) relative to the combination, such as less than about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v), or about 0.01% (w / v).

[0083] In some embodiments, the administration of a combination of the invention results in a beneficial effect, e.g., an additive or synergistic therapeutic effect, e.g., with regard to reducing skin pigmentation or reducing melanin content in a subject. In another embodiment, the administration of a combination of the invention results in further surprising beneficial effects , e.g., fewer side effects, compared to applying only one of the active ingredients used in the combination of the invention.

[0084] Further advantages of employing lower dosages or concentrations of the active ingredients of the combination of the invention include not only the possibility of using smaller concentrations, but also administering them less frequently, while still achieving the desired therapeutic and / or cosmetic effects. Such reduced concentration administered may also lessen the incidence of adverse side effects, such as skin irritation or skin redness, and improve safety during long-term use, minimise systemic toxicity and maintain drug efficacy. This is in accordance with the desires and requirements of the subjects to be treated

[0085] In some embodiments, the sclareolide and ginsenoside R0, or a salt, solvate, derivative, or prodrug thereof is provided at a mole ratio of sclareolide to ginsenoside R0 at about 100:1 to about 1:100. In some embodiments, the mole ratio is about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 12:1, about 10:1, about 8:1, about 6:1, about 5:1, about 4:1, about3:l, about 2:1, about 1.8:1, about 1.5:1, about 1.33:1, about 1.25:1, about 1.2:1, about 1.1:1, or about 1 : 1. In some embodiments, the mole ratio is about 1:1.1, about 1 : 1.2, about 1 : 1.25, about 1:1.33, about 1:1.5, about 1:1.8, about 1:2, about 1:3, about 1:4, about 1:5, about1:6, about 1:8, about 1:10, about 1:12, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, or about 1:100.

[0086] Disclosed herein is a method of inhibiting melanin production in a melanocyte or transfer of melanin in the melanocyte to a keratinocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to inhibit melanin production in the melanocyte or transfer to the keratinocyte.

[0087] Disclosed herein is a method of inhibiting tyrosinase in a melanocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to inhibit tyrosinase in the melanocyte.

[0088] Disclosed herein is a method of lightening or whitening skin in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to the subject.

[0089] Disclosed herein is a method of treating a hyperpigmentation or a hypermelanosis disorder in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to the subject.

[0090] Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, for use in reducing pigmentation in a subject.In some embodiments, the combination is capable of reducing skin pigmentation in a subject.

[0091] In some embodiments, the combination is capable of inhibiting melanin production or transfer in a melanocyte or keratinocyte.

[0092] In some embodiments, the combination is capable of inhibiting tyrosinase in a melanocyte.

[0093] In some embodiments, the combination is capable of lightening or whitening skin in a subject.

[0094] In some embodiments, the combination is capable of treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0095] Disclosed herein is the use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, in the manufacture of a medicament for reducing pigmentation in a subject.

[0096] In one embodiment, there is provided a combination of 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO for use in reducing pigmentation in a subject.

[0097] In one embodiment, there is provided a method of reducing pigmentation in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, for use in treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0098] Disclosed herein is the use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO in the manufacture of a medicament for treating a hyperpigmentation or a hypermelanosis disorder in a subject.

[0099] In one embodiment, there is provided a combination of 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO for use in the treatment of a hyperpigmentation or a hypermelanosis disorder.

[0100] In one embodiment, there is provided a method of treating hyperpigmentation or hypermelanosis in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

[0101] Disclosed herein is a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

[0102] As used herein, the term “combination” refers to a mixture of a composition comprising sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and another composition comprising ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.As such it will be appreciated that the combination of the different composition may be presented as a “kit of parts” for use in the purpose as disclosed herein. The kit may comprise a package where the combination of the compositions are supplied separately for co-admini strati on with instructions for use.

[0103] Disclosed herein is a composition comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

[0104] In some embodiments, the sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, is present in the composition at about 0.001 % (w / v) to about 99 % (w / v), relative to the composition. In other embodiments, the range is about 0.01 % (w / v) to about 99% (w / v). In other embodiments, the range is about 0.1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 5 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 10 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 20 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 30 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 40 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 50 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 60 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 70 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 80 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 90 % (w / v) to about 99 % (w / v).

[0105] In one embodiment, the composition comprises less than about 2 % (w / v), such as less than about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v) or about 0.01% (w / v), of sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, relative to the composition.

[0106] In some embodiments, the ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, is present in the composition at about 0.001 % (w / v) to about 99 % (w / v), relative to the composition. In otherembodiments, the range is about 0.01 % (w / v) to about 99% (w / v). In other embodiments, the range is about 0.1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 1 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 5 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 10 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 20 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 30 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 40 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 50 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 60 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 70 % (w / v) to about 99 % (w / v). In other embodiments, the range is about 80 % (w / v) to about 99 % (w / v). Tn other embodiments, the range is about 90 % (w / v) to about 99 % (w / v).

[0107] In one embodiment, the composition comprises less than 2 % (w / v), such as less than about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v) or about 0.01% (w / v), of ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, relative to the composition.

[0108] In one embodiment, the composition comprises sclareolide, or a salt, solvate, derivative or prodrug thereof, or botanical extract comprising sclareolide, at a concentration of less than 1 mM, less than 500 pM, less than 100 pM, less than 50 pM, less than 10 pM, less than 5 pM, less than 1 pM, less than 500 nM, less than 300 nM, less than 100 nM, such as between 50 nM to 1 mM, between 50 nM to 500 pM, between 50 nM and 100 pM, between 50 nM and 50 pM, between 50 nM and 10 pM, between 50 nM and 5 pM, between 50 nM and 1 pM, between 50 nM and 500 nM, between 50 nM and 300 nM, or between 50 nM and 100 nM.

[0109] In one embodiment, the composition comprises ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or botanical extract comprising ginsenoside R0, at a concentration of less than 1 mM, less than 500 pM, less than 100 pM, less than 50 pM, less than 10 pM, less than 5 pM, less than 1 pM, less than 500 nM, less than 300 nM, less than 100 nM, such as between 50 nM to 1 mM, between 50 nM to 500 pM, between 50 nM and 100 pM, between 50 nM and 50 pM, between 50 nM and 10 pM, between50 nM and 5 LIM. between 50 nM and 1 pM, between 50 nM and 500 nM, between 50 nM and 300 nM, or between 50 nM and 100 nM.

[0110] In some embodiments, the composition comprises sclareolide and ginsenoside with a mole ratio of sclareolide to ginsenoside R0 at about 100:1 to about 1:100. In some embodiments, the mole ratio is about 100:1, about 90:1, about 80:1, about 70:1, about 60:1, about 50:1, about 40:1, about 30:1, about 20:1, about 15:1, about 12:1, about 10:1, about 8:1, about 6:1, about 5:1, about4:l, about 3:1, about2:l, about 1.8:1, about 1.5:1, about 1.33:1, about 1.25:1, about 1.2:1, about 1.1:1, or about 1:1. In some embodiments, the mole ratio is about 1:1.1, about 1:1.2, about 1:1.25, about 1:1.33, about 1:1.5, about 1:1.8, about 1 :2, about 1 :3, about 1 :4, about 1 :5, about 1 :6, about 1 :8, about 1 :10, about 1:12, about 1:15, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:70, about 1:80, about 1:90, or about 1:100.

[0111] In some embodiments, the composition has a pH value of about 3.5 to about 6.5, such as about 3.5, about 4.0, about 4.5, about 5.0, about 5.5, about 6.0, about 6.5, about 7.0, or about 7.5.

[0112] In some embodiments, the composition further comprises a tyrosinase inhibitor, a corticosteroid, a hydroquinone, a melanin synthesis inhibitor, a melanin transfer inhibitor, or a combination thereof.

[0113] Examples of tyrosinase inhibitor include, but are not limited to, tyrosinase-IN-25, tyrosinase-IN-46, tyrosinase-IN-1, tyrosinase-IN-2, tyrosinase-IN-3, tyrosinase-IN-4, persiconin, broussonin C, methyl rosmarinate, kurarinol, isolindleyin, swertiajaponin, 2-methoxycinnamic acid, dihydromorin, alashinol G, tetrapeptide-30, decapeptide- 12, (-)-2-fluorodeoxyarbutin, glycolic acid, tropinone, benzoyloxypaeoniflorin, (R)-trolox, neorauflavane, dihydroaltenuene B, (+)-2-fluorodeoxyarbutin, melanocin A, melanocin B, melanocin C, kazinol U, 4-butylresorcinol, litchinol B, 7-methoxy obtusifolin, allamandicin, chimonanthine, pseudostellarin G, pedalitin, sanggenon O, Mirificin, kojic acid, Viscumneoside III, Glycolic acid-d2, Hordenine sulfate, glabrene, AP736, chlorophorin, gnetol, oxyresveratrol, MHY908, Taraxinic acid, Dodoviscin A, 3-(2,4-Dihydroxyphenyl)propanoic acid, Oxyresveratrol 3'-O-P-D-glucopyranoside, D-Tyrosine, 5-Methylresorcinol monohydrate, (-)-Isodocarpin, 6-Acetylnimbandiol, p-tocopherol, and 6-Hydroxy kaempferol.

[0114] Examples of melanin synthesis inhibitor include, but are not limited to, melanocin A, melanocin B, melanocin C, tyrosinase-IN-25, tyrosinase-IN-46, tyrosinase-IN-1, tyrosinase-IN-2, tyrosinase-IN-3, tyrosinase-IN-4, swertiajaponin, decapeptide- 12, glycolic acid, neorauflavane, Pseudostellarin G, kojic acid, Glycolic acid-d2, Hordenine sulfate, chlorophorin, gnetol, MHY908, Dodoviscin A, Phenylthiourea, D-Tyrosine, 6-Acetylnimbandiol, (-)-Isodocarpin, hydroquinone, arbutin, azelaic acid, vitamin C, ascorbic acid, tranexamic acid, niacinamide, retinoid, tretinoin, retinol, liquirtin, 4-0-methylsappanol, velutin, tolprocarb, -tocopherol, epirosmanol, cetyl tranexamate hydrochloride, dehydroglyasperin D, 2-ethoxybenzamide, nonapeptide- 1 acetate salt, and cysteamine.

[0115] Examples of melanin transfer inhibitor include, but are not limited to, swertiajaponin, niacinamide, lectins, metformin, PAR-2 inhibitors, decanoic acid, nonapeptide- 1 acetate salt, nonapeptide- 1, paeonol, isoliquiritigenin, chitosan, retinoid, tretinoid, and retinol.

[0116] Examples of corticosteroids include, but are not limited to, fluocinolone acetonide, hydrocortisone, mometasone furoate, betamethasone, desonide, clobetasol propionate, and triamcinolone acetonide.

[0117] In some embodiments, the composition is a pharmaceutical composition. In a suitable embodiment, the composition may be formulated as a topical composition. In another embodiment, the composition may be formulated as a transdermal composition.

[0118] In some embodiments, the composition is a cosmetic or a non-therapeutic composition. The composition may be a skincare composition, such as a skin whitening or lightening composition.

[0119] The composition may include a cosmetic vehicle. Such vehicles may take the form of any known in the art suitable for application to the skin. These vehicles may include water; vegetable oils; mineral oils, esters such as octyl palmitate, isopropyl myristateand isopropyl palmitate; ethers such as dicapryl ether and dimethyl isosorbide; alcohols such as ethanol and isopropanol; fatty alcohols such as cetyl alcohol, cetearyl alcohol, stearyl alcohol and behenyl alcohol; isoparaffins such as isooctane, isododecane and isohexadecane; silicone oils such as cyclomethicone, dimethicone, dimethicone crosspolymer, polysiloxanes and their derivatives preferably organomodified derivatives, hydrocarbon oils such as mineral oil, petrolatum, isoeicosane and polyisobutene; polyols such as propylene glycol, glycerin, butylene glycol, pentylene glycol and hexylene glycol; waxes such as beeswax and botanical waxes; and mixtures of the foregoing.

[0120] The present composition can be made into any suitable product form, such as an aerosol, cake, cream, ointment, emulsion, essence, gel, lotion, paste, patch, pencil, serum, towelette, mask, spray and stick. If desired, the composition can be formulated into make-up cosmetics or soaps.

[0121] The present composition may further comprise one or more of the following additional ingredients: anaesthetics, anti-allergenics, antimicrobials, antifungals, antiinflammatories, antiseptics, chelating agents, colorants, depigmenting agents, emollients, exfoliants, fragrances, emulsifiers, humectants, insect repellents, lubricants, moisturizers, pharmaceutical agents, preservatives, skin penetration enhancers, skin protectants, stabilizers, sunscreens, surfactants, thickeners, viscosity modifiers, and vitamins.

[0122] In some embodiments, the composition comprising can be applied and left on the skin of the subject for at least 1, 2, 3, 4, 5, 6, 7 or 8 hours or more without removing or washing off the composition. In some embodiment, the composition can be re-applied on the skin of the subject every 8 hours with or without washing off the composition present on the skin of the subject.

[0123] The composition may be applied directly to the skin by any suitable means, including, by hand, with a spatula, scoop or spoon, spraying, dusting, painting and pouring. The composition is preferably massaged into the hair or skin. While some oils, such as rosehip, jojoba, argan, olive, coconut, and sunflower oils, are rapidly absorbed into thehair (about 2-4 hours), others, such as avocado, castor, evening primrose and macadamia nut oils, are absorbed more slowly, (about 6-8 hours). Thus, while some benefit may be obtained while the oil-based compositions are left on for 2-4 hours, oil-based compositions are preferably left on for at least about 6-8 hours, or longer, such as 8-12 hours, and may conveniently be left on overnight before washing out. Compositions comprising solid components may be mixed with a suitable cosmetically acceptable carrier, in the sense of being compatible with the other ingredients of the composition and not injurious to the subject. Some examples of a suitable carrier include water, oils, waxes, and butters extracted from seeds, such as cocoa butter, shea butter and mango butter and mixtures thereof. The solid components may be mixed with a suitable carrier before applying to the skin, and then massaged in. Such compositions may be left in for any suitable period of time, and in some embodiments, are preferably left on for about 10-40 minutes, such as about 15-30 minutes, before washing or rinsing out. A treatment process may be performed as a one-off treatment or may be used on a regular basis over a period of time, for example, the composition may be applied daily, once or twice weekly, fortnightly or monthly.

[0124] The term “excipient” as used herein encompasses carriers and diluents, meaning a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material involved in carrying or transporting the composition comprising sclareolide and / or ginsenoside RO, and may be delivered to the target site.

[0125] The carrier of the topical composition may further comprise one or more ingredients selected from the group consisting of emollients, propellants, solvents, humectants, thickeners, powders, fragrances and pH-adjusting additive.

[0126] The amount of emollient in the topical composition is typically about 5% to about 95%. Non-limiting examples of emollients include stearyl alcohol, glyceryl monoricinoleate, glyceryl monostearate, propane- 1,2-diol, butane-l,3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, iso-butyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, polydimethylsiloxane, di-n-butyl sebacate, iso-propyl myristate, iso-propyl palmitate, iso-propyl stearate, butyl stearate, polyethylene glycol, triethylene glycol,lanolin, sesame oil, coconut oil, arachis oil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof.

[0127] Oils include cannabidiol oil and various plant derived oils containing cannabidiol, such as, hempseed oil, Echinacea purpurea Echinacea angustifolia, Acmella oleracea, Helichrysum umbraculigerum, Radula marginata, and the like. The cannabidiol isolated from such plants or made synthetically may be formulated with an oil such as, for example, olive oil, grapeseed oil, tea tree oil, almond oil, avocado oil, sesame oil, evening primrose oil, sunflower oil, kukui nut oil, jojoba oil, walnut oil, peanut oil, pecan oil, macadamia nut oil, coconut oil, and the like and combinations thereof.

[0128] The amount of propellant in the topical composition is typically about 5% to about 95%. Non-limiting examples of propellants include propane, butane, iso-butane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof.

[0129] The amount of solvent in the topical composition is typically about 5% to about 95%. Non-limiting examples of solvents include water, ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulphoxide, dimethyl formamide, tetrahydrofuran, and combinations thereof. Preferred solvents include ethyl alcohol.

[0130] The amount of humectant in the topical composition is typically about 5% to about 95%. Non-limiting examples of humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. Preferred humectants include glycerin.

[0131] The amount of thickener in the topical composition is typically 0% to about 95%. Nonlimiting examples of thickeners include xanthan gum, guar hum / hydroxypropyl guar, cellulose derivatives (such as hydroxyethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and carboxymethylcellulose), alginates, pectin, agar, starch derivatives, synthetic polymer thickeners, stearic acid, palmitic acid, waxes, sheabutter, cocoa butter, mineral thickeners, hydroxypropyl starch phosphate, PEG- 150 distearate, PEG-150 pentaerythrityl tetrastearate, lauryl glucoside, sodium chloride, sodium sulfate, sodium polystyrene sulfonate, protein-based thickeners (such as collagen, hydrolyzed collagen, and gelatin), and silicone-based thickeners.

[0132] The amount of powder in the topical composition is typically 0% to about 95%. Nonlimiting examples of powders include chalk, talc, fullers earth, kaolin, starch, gums, colloidal silicon dioxide, sodium polyacrylate, tetra alkyl ammonium smectites, trialkyl aryl ammonium smectites, chemically modified magnesium aluminum silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and combinations thereof.

[0133] The amount of fragrances in the topical composition is typically about 0.001% to about 0.5%.

[0134] Ointments are compositions in which oil and water are provided in a ratio of from 7: 1 to 2: 1, from 5: l to 3: l, or4: 1 Ointments are generally formulated using oils, waxes, water, alcohols, petroleum products, water, and other agents to prepare compositions with various viscosities and solvent properties. Commonly used compositions include oleaginous base (White Ointment), absorption base, W / O emulsion base (Cold Cream type base), O / W emulsion base (Hydrophilic Ointment), water soluble base, in addition to others. These preparations are used to dissolve or suspend substances or products with medicinal or cosmetic value.

[0135] Lotions are low- to medium-viscosity topical preparations. Most lotions are oil-in-water emulsions containing an emulsifier such as cetyl alcohol to prevent separation of these two phases. Lotions can include fragrances, glycerol, petroleum jelly, dyes, preservatives, proteins and stabilizing agents.

[0136] In some embodiments, the composition as disclosed herein further comprises an emulsifier. The emulsifier may be selected from emulsifying ointment BP, isopropylmyristate, hydroxyethylcellulose, glycerol, phenoxyethanol, propylene glycol, and a combination thereof.

[0137] In some embodiments, the composition as disclosed herein is formulated with a delivery vehicle. For example, in a suitable embodiment, the composition as disclosed herein is complexed with or encapulsated within a liposome.

[0138] By way of further example, suitable delivery vehicles include those selected from the group consisting of liposomes, amphoteric and cationic liposomes, niosomes, lipophilic formulations, aqueous-alcoholic solutions, hydrophilic formulations, water-in-oil nanoemulsions, nanoparticles of various sizes, microparticles of various sizes, polystyrene microspheres of various sizes, titanium dioxide particles and solid lipid particles.

[0139] Suitably liposomes employed in the compositions or methods of the invention may be ones that have a particular affinity for fibroblast, smooth muscle cells, endothelial cells, or any other suitable target cell types. By way of example suitable liposomes include, amphoteric and cationic liposomes and niosomes

[0140] As used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (or).

[0141] As used in this application, the singular form "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the term "an agent" includes a plurality of agents, including mixtures thereof.

[0142] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.The reference in this specification to any prior publication (or information derived from it), or to any matter which is known, is not, and should not be taken as an acknowledgment or admission or any form of suggestion that that prior publication (or information derived from it) or known matter forms part of the common general knowledge in the field of endeavor to which this specification relates.

[0143] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications, which fall within the spirit and scope. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.

[0144] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0145] Certain embodiments of the invention will now be described with reference to the following examples which are intended for the purpose of illustration only and are not intended to limit the scope of the generality hereinbefore described.

[0146] EXAMPLES

[0147] Methods

[0148] The sclareolide and / or ginsenoside RO were dissolved in DMSO at a desired concentration. The RHPE samples were pre-equilibrated for 24 h in EPI-100-NMM-113 media (MatTek) before any treatment. Once equilibrated, the RHPE samples were treated with sclareolide, ginsenoside RO or a combination thereof for a total of 14 days. For melanin quantification, the RHPE samples were incubated in 1% sodium bicarbonate for 30 min, followed by incubation with Solvable™ at 60 °C for at least 16 hours. Melanin levels were determined using a spectrophotometer at 405 nm.Sclareolide 100 nM, and ginsenoside RO 10 pM, in combination, showed a clear antipigmentation effect under baseline NO UVB condition after 14 days (treated every 2 days) in the 3D RHPE samples compared with the vehicle 0.5% DMSO control samples and single actives (see Figure 1) Melanin quantification normalized by protein showed a 23% reduction in the samples (n=3) treated with combination of 100 nM sclareolide and 10 pM ginsenoside R0 compared with the untreated controls (0.5% DMSO). Single compound treatment of 100 nM sclareolide or 10 pM ginsenoside R0 showed 6% and 3% melanin reduction, respectively. Dotted horizontal lines represent threshold of 15% and 60%, as labeled. Solid black horizontal solid line represents mean DMSO control, (one-way ANOVA, **p < 0.01, ***p < 0.001). Error bars represent mean ± SEM.

[0149] Results: Combination of lOOnM sclareolide and lOpM ginsenoside R0 shows robust anti -pigmentation effects compared to single actives in the 3D Reconstructed Human Pigmented Epidermal (RHPE) model under baseline NO UVB-irradiated condition.

Claims

CLAIMS1. A method of reducing skin pigmentation in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

2. The method of claim 1, wherein the subject is a mammalian subject, such as a human subject.

3. The method of claims 1 or 2, wherein the combination lightens the subject’s pigmented skin.

4. The method of claim 3, wherein the subject is suffering from a hyperpigmentation or a hypermelanosis disorder.

5. The method of any one of claims 1 to 4, wherein the sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, is provided at a concentration of less than 1 mM (such as between 50 nM to 1 mM).

6. The method of any one of claims 1 to 5, wherein the ginsenoside RO, a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, is provided at a concentration of less than 1 mM (such as between 50 nM to ImM).

7. The method of any one of claims 1 to 6, wherein the sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, is provided at a concentration of less than about 2 % (w / v) relative to the combination.

8. The method of claims 1 to 7, wherein the ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, is provided at a concentration of less than about 2 % (w / v) relative to the combination.

9. The method of any one of claims 1 to 8, wherein the sclareolide and ginsenoside RO, or a salt, solvate, derivative, or prodrug thereof is provided at a mole ratio of sclareolide to ginsenoside RO at about 100: 1 to about 1 : 100.

10. The method of any one of claims 1 to 9, wherein the method is a non-therapeutic or cosmetic method.

11. The method of any one of claims 1 to 10, wherein the method comprises topically or transdermally administering the combination (either concurrently or sequentially) to the subject.

12. A method of inhibiting melanin production in a melanocyte or transfer of melanin in the melanocyte to a keratinocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to inhibit melanin production in the melanocyte or transfer to the keratinocyte.

13. A method of inhibiting tyrosinase in a melanocyte, the method comprising contacting the melanocyte with a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to inhibit tyrosinase in the melanocyte.

14. A method of lightening or whitening skin in a subject, the method comprising administering a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside R0, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside R0, to the subject.

15. A method of treating a hyperpigmentation or a hypermelanosis disorder in a subject, the method comprising administering a combination comprising 1)sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, to the subject.

16. A combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, for use in reducing pigmentation in a subject.

17. A combination comprising 1) sclareolide, ora salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, for use in the treatment of a hyperpigmentation or a hypermelanosis disorder.

18. Use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO, in the manufacture of a medicament for reducing pigmentation in a subject.

19. Use of a combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO in the manufacture of a medicament for treating a hyperpigmentation or a hypermelanosis disorder in a subject.

20. A combination comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

21. A composition comprising 1) sclareolide, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising sclareolide, and 2) ginsenoside RO, ora salt, solvate, derivative or prodrug thereof, or a botanical extract comprising ginsenoside RO.

22. The composition of claim 21, wherein the composition comprises less than 2 % (w / v) of sclareolide, or a salt, solvate, derivative or prodrug thereof, relative to the composition.

23. The composition of claims 21 or 22, wherein the composition comprises less than 2 % (w / v) of ginsenoside RO, or a salt, solvate, derivative or prodrug thereof, relative to the composition.

24. The composition of any one of claims 21 to 23, wherein the mole ratio of sclareolide to ginsenoside RO, or a salt, solvate, derivative, or prodrug thereof is about 100:1 to about 1:100.

25. The composition of any one of claims 21 to 24, wherein the pH value is about 3.5 to about 7.5.

26. The composition of any one of claims 21 to 25, wherein the composition further comprises a tyrosinase inhibitor, a corticosteroid, a hydroquinone, a melanin synthesis inhibitor, a melanin transfer inhibitor, or a combination thereof.

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