Method of promoting tropoelastin synthesis

Abstract

The invention relates to methods of skincare. In particular, the present specification teaches a method of promoting tropoelastin synthesis comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to a subject, as well as compositions as defined herein.

Description

[0001] METHOD OF PROMOTING TROPOELASTIN SYNTHESIS

[0002] Field of Invention

[0003] The invention relates to methods of skincare. In particular, the present specification teaches a method of promoting tropoclastin synthesis in a subject as well as compositions as defined herein.

[0004] Background

[0005] Elastic fibres are an integral component of the extracellular matrix (ECM) involved in giving connective tissues elasticity and mechanical resilience. They comprise multiple components, including a crosslinked elastin core surrounded by a mantle of microfibrils made up of mainly fibrillin. Elastic fibres are designed to be durable enough to last a lifetime, with de-novo synthesis occurring during neonatal phase and rarely in adults. Precisely because of this, however, elastic fibres are also prone to accumulate damages over time. Elastic fibres isolated from older individuals are more susceptible to enzymatic attacks, which may be aggravated by extrinsic factors including UV radiation and lifestyle choices. In addition, elastic fibres and other long-living proteins are also susceptible to non-cnzymatic glycation, which further alters their mechanical property.

[0006] Due to this increased degradation and insufficient repairs of damaged elastic fibres, the net result contributes to an aged skin where there is a loss of elasticity and sagging. The proper re- stimulation of elastin synthesis is therefore a potential goal in anti-aging product formulation. However, most anti-ageing strategies are centred on collagen and rarely on elastin, save for retinol which was shown to boost the production of both. Thus, there remains space for innovation, as consumers are constantly seeking novel ingredients that are safer and potentially more efficacious.

[0007] It would be desirable to overcome or ameliorate at least one of the above-described problems, or at least to provide a useful alternative. Summary

[0008] Disclosed herein is a method of promoting tropoelastin synthesis in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0009] Disclosed herein is a method of promoting tropoelastin synthesis in a dermal fibroblast, the method comprising contacting the dermal fibroblast with berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine.

[0010] Disclosed herein is a method of improving the appearance of aging or sagging skin in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0011] Disclosed herein is a method of improving skin firmness and / or elasticity in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0012] Disclosed herein is a method of promoting a youthful appearance in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0013] Disclosed herein is berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine for use in promoting tropoelastin synthesis in a subject.

[0014] Disclosed herein is the use of berberine, or a salt, solvate, derivative or prod rug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for promoting tropoelastin synthesis in a subject.

[0015] Disclosed herein is the use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for treating a cutaneous wound. Disclosed herein is the use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for treating a disease associated with a loss of elastin fibres.

[0016] Disclosed herein is a composition comprising berberine, or a salt, solvate, derivative or prodrug thereof, or comprising a botanical extract comprising berberine.

[0017] Brief Description of Drawings

[0018] Embodiments of the present invention are hereafter described, by way of non-limiting example only, with reference to the accompanying drawings in which:

[0019] Figure 1. Berberine hydrogen sulphate induces tropoclastin synthesis in dermal fibroblasts. Fold induction of tropoelastin in dermal fibroblasts treated with either benchmark compound TGF-p l . or berberine hydrogen sulphate (BBR-HS), relative to the respective vehicle controls. NT, non-treated control.

[0020] Figure 2: Other salt forms of berberine can similarly induce tropoelastin. Other salt forms of Berberine can similarly induce elastin. DAPI nuclei counts (top row) and corresponding elastin quantification (bottom row) from dermal fibroblasts treated with the indicated concentrations of berberine, berberine chloride, berberine hydrogen sulphate and berberine sulfate from different vendors (TargetMol or Selleck Chemicals).

[0021] Figure 3: Comparison of Berberine hydrogen sulphate’s efficacy against cosmetic benchmarks. Tropoelastin quantification from the supernatant of dermal fibroblasts treated with the indicated concentrations of berberine hydrogen sulphate (BHS), or other known cosmetic ingredients. The black dash line represents baseline of vehicle control set to 1.

[0022] Figure 4 shows the structure of berberine hydrogen sulphate.

[0023] Figure 5: (A) Representative data showing immunofluorescence staining for elastin (green) and nuclei (red) in ex vivo human skin treated with either vehicle control or BBR-HS in the cell culture medium. (B) Quantification data for samples in (A). A total of 9 different fields of view were imaged in 3 replicates per sample and quantified as % coverage of staining. Error bars are mean ± SD. ** p<0.01.

[0024] Figure 6: Tropoelastin fold-induction relative to non-treated control from the supernatant of dermal fibroblasts treated with increasing concentrations of Berberis vulgaris root / bark botanical extract containing BBR. All error bars are mean ± SEM.

[0025] Figure 7: Different berberine-containing botanical extracts can similarly induce tropoelastin expression. Tropoelastin fold-induction relative to non-treated control from the supernatant of dermal fibroblasts treated with different concentrations of 3 botanical extracts. Error bars are ± SEM from at least 3 independent experiments.

[0026] Detailed Description

[0027] The present specification teaches a method of promoting tropoelastin synthesis in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0028] As used herein, the term “tropoelastin” refers to the precursor of elastin, wherein the elastin is a component of the extracellular matrix (ECM) and provides tissues with elasticity and flexibility. The elastin fibres are formed through multimerization and crosslinking of tropoelastin monomers in the presence of elastic proteins including fibrillins, fibulins, and lysyl oxsidases. During development, cells, such as fibroblast, smooth muscle cells, and endothelial cells secrete soluble monomers of tropoelastin, which are then assembled by the enzyme lysyl oxidase within the ECM to form elastin fibres, which are insoluble polymers with high stability. The process of elastin fibre formation is also known as “elastic fibre formation”.

[0029] Without being bound by theory, the inventors have identified the heteropentacyclic alkaloid, berberine, as a natural ingredient that induces tropoelastin synthesis in dermal fibroblasts. The in vitro data showed that salt forms of berberine (i.e. berberine hydrogen sulphate, berberine hydrochloride and berberine sulfate), as well as botanical extracts containing them (Berberis vulgaris, Phellodendron amurense, Coptis chinensis), are able to induce tropoelastin synthesis with minimal toxicity at low treatment dosage.

[0030] The botanical extract may be any botanical extract that contains berberine. For example, the botanical extract can be a botanical extract from Berberis vulgaris, Berberis aristata, Coptis chinensis, Coptis japonic a, Phellodendron amurense bark extract, etc.

[0031] In one embodiment, the salt of berberine is berberine hydrogen sulphate, berberine hydrochloride or berberine sulfate. Tn one embodiment, the salt of berberine has a structure

[0032] As used herein, the term “derivative” refers to a chemical compound that is derived from berberine, comprising substitution and / or removal of a functional group through a chemical reaction.

[0033] In one embodiment, the derivative of berberine is a protoberberine alkaloid, optionally wherein the protoberberine alkaloid is epiberberine, palmatine, coptisine, jatrorrhizine or berberrubine.

[0034] As used herein, the term “prodrug” refers a compound which is in a form that requires activation, such as through enzymatic and / or biotransformation, before the berberine can have a significant therapeutic effect on the subject in which the inhibitor is administered to. The biotransformation process includes metabolism process of the subject that the agent or the inhibitor is administered to.

[0035] In one embodiment, berberine may act as a prodrug to the berberine metabolite. Examples of berberine metabolites include but arc not limited to berberrubine, thalifendine, demethyleneberberine, jatrorrhizine, palmatine, columbamine and dihydroberberine. 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 polymorphs of 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, cmbonic (pamoic), mcthancsulfonic, cthancsulfonic, bcnzcncsulfonic, pantothenic, toluenesulfonic, 2-hydroxyethanesulfonic, sulfanilic, cyclohcxylaminosulfonic, algcnic, 3-hydroxybutyric, galactaric and galacturonic acid.

[0036] As used herein, the term “botanical extract” refers to concentrated plant materials or naturally occurring compounds present in a plant material that is isolated using methods known in the art, such as immersion in a suitable solvent, boiling, filtration, distillation, pulverization, dilution, freezing etc. The isolated compound such as berberine or its salts and derivatives may be further fabricated into a composition that is suitable for promoting the tropoelastin and elastin production in a subject.

[0037] In some embodiments, berberine or its salts and / or derivatives may be extracted from different parts of the plant, including but not limited to flower, seed, rhizome, root, bark, leaf, or fruit.

[0038] In some embodiments, berberine or its salts and / or derivatives may be extracted from the plant of Berberis vulgaris, Phellodendron amurense, or Coptis chinensis using a suitable solvent. Examples for suitable solvent for extracting berberine from botanical sources include but are not limited to water, ethanol, methanol, hexane, acetone, chloroform, dichloromethane, petroleum ether, ethyl acetate, and 1 ,3-butylene glycol. In some preferred embodiments, the solvents are water, ethanol, or 1,3-butylene glycol. In some embodiments, different solvents may be used simultaneously or consequently in any combination of compounds or concentrations.

[0039] In one embodiment, berberine may be extracted from Berberis vulgaris using water as a solvent. The water may be about 25% to about 100%, such as about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% volume to weight ratio of the plant material.

[0040] In one embodiment, berberine may be extracted from Berberis vulgaris using ethanol as a solvent. The ethanol may be about 25% to about 100%, such as about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% volume to weight ratio of the plant material.

[0041] In one embodiment, berberine may be extracted from Phellodendron amurense using 1,3-butylene glycol as a solvent. The 1,3-butylene glycol may be about 25% to about 100%, such as about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% volume to weight ratio of the plant material.

[0042] In one embodiment, berberine may be extracted from Coptis chinensis using 1,3- butylene glycol as a solvent. The 1,3-butylene glycol may be about 25% to about 100%, such as about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% volume to weight ratio of the plant material.

[0043] The method may comprise promoting elastin synthesis in dermal fibroblast of a skin tissue in the subject.

[0044] The method may comprise promoting tropoelastin synthesis in a fibroblast of a subject, such as fibroblast from the lung, blood vessel or intestine. This may be useful to treat any disease linked a loss of elastin fibres including but not limited to lung emphysema or vascular aneurysm.

[0045] In some embodiments, a subject may be suffering from a condition associated with loss of elastin fibres and / or loss of tropoelastin monomers, such as aneurysms, atherosclerosis and loss of skin elasticity. In some embodiments, the loss of elastin may be congenital (genetically-linked) or acquired. In one embodiment, the loss of elastin is caused by genetic variation in the elastin ELN) gene. The disease and / or condition a subject may be suffering from includes but is not limited to autosomal dominant cutis laxa (ADCL), supravalvar aortic stenosis (SVAS), Williams Beuren Syndrome (WBS) and mid-dermal elastolysis (MDE).

[0046] In some embodiments, a subject may be suffering from a condition associated with UV- induced or UV -related disease, such as photoaging or dark spots.

[0047] In some embodiments, a subject may be suffering from a cutaneous wound. The term “cutaneous wound” refers to an injury or a disruption of normal anatomic structure and function of the skin that occurs in a subject. The cause of cutaneous wound include but are not limited to abrasion, burn, trauma, surgical incision, cut, scratch, needle-prick and long-term wound or lesion.

[0048] The method may comprise promoting tropoelastin synthesis in dermal fibroblasts of a skin tissue in the subject.

[0049] The method may comprise promoting elastin synthesis in dermal fibroblasts of a skin tissue in the subject.

[0050] The method may comprise promoting elastic fibre formation in demal fibroblast of a skin tissue in the subject.

[0051] As used herein, the tern “promoting”, “increasing”, “improving”, and “enhancing” may be used interchangeably when referring to promoting tropoelastin synthesis. The berberine may increase of tropoelastin synthesis in a subject. The increase may be, for example, an increase of at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%x 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% increase or any increase between 5-100% as compared to a reference level.

[0052] As used herein, the term “reference” refers to a subject that is not administered with a composition comprising berberine for promoting tropoelastin synthesis or before a subject is administered with a composition comprising berberine for promoting tropoelastin synthesis.

[0053] In one embodiment, the method improves the appearance of aging or sagging skin in the subject. In another embodiment, the method improves skin firmness and / or elasticity in a subject. In another embodiment, the method promotes a youthful appearance in a subject. The method may also increase skin volume or decrease symptoms of photoaging.

[0054] In one embodiment, the method to measure skin firmness and / or elasticity is skin cutometry using a suitable device, such as a skin cutometer. The method is disclosed in patent US7556605B2, entitled “Methods for determining elastic and viscoelastic properties of skin” is hereby incorporated in its entirety by reference.

[0055] As used herein, the term “promoting”, “increasing”, “improving”, and “enhancing” may be used interchangeably when referring to improving skin firmness and / or elasticity in a subject. The berberine may increase or improve skin firmness and / or elasticity in a subject. The increase may be, for example, an increase of at least about 1%, about 2%, about 3%, about 5%, about 7.5%, about 10%, at least about 15%, at least about 20%, 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% increase or any increase between 1-100% as compared to a reference level. In one embodiment, the method to assess appearance of aging or sagging skin and / or youthful appearance 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 fullface 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 puffiness, 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 skin sagging, 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.

[0056] The berberine may improve or decrease the skin sagging appearance, decrease skin aging appearance or promote a youthful appearance of the skin in a subject. The berberine may increase the score of the skin in a subject. The improvement or decrease of aging or sagging appearance may be measured from the presence of aging features (such as fine lines, wrinkles, dark spots, etc) on the image of the face of the subject, and may be measured the frequency and area coverage of the aging feature on the image of the face of the subject. The higher the frequency and / or the larger the area coverage of the aging features is indicative that the subject has an aged skin, and may be assigned a lower score compared to a reference.

[0057] The improvement in the score may be, for example, an increase of at least about 0.1%, about 0.5%, about 1%, about 2%, about 3%, about 5%, about 7.5%, about 10%, at least about 15%, at least about 20%, 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% increase or any increase between 0.1-100% as compared to a reference level. The improvement in the score may be, for example, about 1 times, about 1.5 times, about 2 times, about 3 times, about 4 times, about 5 times, about 8 times, about 10 times, about 15 times, or about 20 times as compared to a reference level.

[0058] In one embodiment, the berberine, or a salt, solvate, derivative or prodrug thereof, or botanical extract comprising berberine, is provided at a dose of less than 15 u M (or less than 4.95 pg / ml), less than 10 pM (or less than 3.3 pg / ml), less than 5 pM (or less than 1.65 pg / ml) or less than 1 pM (or less than O.33pg / ml), such as between 100 nM to 15 pM, between 200 nM to 15 pM, between 300 nM to 1 pM, between 400 nM to 15 pM, between 500 nM to 15 pM, between 100 nM to 10 pM, between 200 nM to 10 pM, between 300 nM to 10 pM, between 400 nM to 10 pM, between 500 nM to 10 pM, between 100 nM to 5 pM, between 200 nM to 5 pM, between 300 nM to 5 pM, between 400 nM to 5 pM, between 500 nM to 5 pM, between 100 nM to 1 pM, between 200 nM to 1 pM, between 300 nM to 1 pM, between 400 nM to 1 pM or between 500 nM to 1 pM.

[0059] The method may be a non-therapeutic or cosmetic method.

[0060] 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 be a mammalian subject. The “subject” may include a person, a patient or individual, and may be of any age or gender.

[0061] 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. The term “administering” refers to contacting, applying, or providing berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0062] In one embodiment, the method comprises topically and / or transdermally administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

[0063] In one embodiment, the method further comprises administering retinol to the subject.

[0064] In one embodiment, the method further comprises administering TGF-pi to the subject.

[0065] By “effective amount”, in the context of treating or preventing a condition is meant the administration of an amount of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine 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.

[0066] The present specification also teaches a method of promoting tropoelastin synthesis in a dermal fibroblast, the method comprising contacting the dermal fibroblast w'ith berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine.

[0067] Disclosed herein is a method of improving the appearance of aging or sagging skin in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject. Disclosed herein is berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine for use in promoting tropoelastin synthesis in a subject.

[0068] Disclosed herein is the use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for promoting tropoelastin synthesis in a subject.

[0069] Disclosed herein is the use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for treating a cutaneous wound. The medicament may promote de novo synthesis of tropoelastin within the wound site.

[0070] Disclosed herein is a composition comprising berberine, or a salt, solvate, derivative or prodrug thereof, or comprising a botanical extract comprising berberine.

[0071] The composition may be a topical composition. The present invention may comprise berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine for topical application to skin for a period of time and in an amount sufficient to improve the appearance of aging or sagging skin.

[0072] In some embodiments, the composition may be formulated into a cosmetic formulation. In some embodiments, the composition may be formulated into a pharmaceutical formulation.

[0073] The composition may comprise less than 0.0005% (w / v), less than 0.0001% (w / v), less than 0.00005% (w / v), less than 0.0004% (w / v), less than 0.0003% (w / v), less than 0.0002% (w / v) or less than 0.00001% (w / v) of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine.

[0074] 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 'ater; vegetable oils; mineral oils; esters such as octyl palmitate, isopropyl myristate and 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.

[0075] 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. The composition of the invention may be in any pharmaceutical form normally used for topical application, in particular in the form of an aqueous, aqucous-alcoholic or oily solution, an oil-in-watcr or water-in-oil or multiple emulsion, an aqueous or oily gel, a liquid, pasty or solid anhydrous product, a dispersion of oil in an aqueous phase with the aid of spherules, these spherules possibly being polymeric nanoparticles such as nanospheres and nanocapsules or better still lipid vesicles of ionic and / or non-ionic type.

[0076] This composition may be relatively fluid and have the appearance of a white, clear, or colored cream, an ointment, a milk, a lotion, a serum, a cream, a paste, a foam, a gel, an oil or a spray. It may optionally be applied to the skin in aerosol form. It may also be in solid form and, for example, in the form of a stick. It can be used as a care product and / or as a make-up product.

[0077] In some embodiments, the berberine can be supplemented with any of the additives discussed below and can be incorporated into creams, lotions, salves, liniments, ointments, gels, pastes, tonics, tinctures, unguents, soaps, orals, pills, tablets, capsules, and lip balms discussed below. Thus, the form of the compositions of the invention is not limited. The present composition may further comprise one or more of the following additional ingredients: anesthetics, 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.

[0078] 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.

[0079] 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 the hair (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 w ashing 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.

[0080] The term “carrier” as used herein encompasses carriers, excipients, 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 berberine compound across a tissue layer such as the stratum comeum or stratum spinosum, and may be delivered to the target site.

[0081] The carrier of the berberine in a 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. Typical carriers for the topical compositions include water, alcohols, aloe vcra gel, allantoin, glycerin, vitamin A and E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbidc, combinations thereof, and the like. Preferred carriers include propylene glycol, dimethyl isosorbide, and water.

[0082] Creams refer to semi- solid emulsions of oil and water in approximately equal proportions. They are divided into two types: oil-in-water (O / W) creams, composed of small droplets of oil dispersed in a continuous phase; and water-in-oil (W / O) creams, composed of small droplets of water dispersed in a continuous oily phase. Creams can provide a barrier to protect the skin. This may be a physical barrier or a chemical barrier as with UV-absorbing compounds. To aid in the retention of moisture (especially water- in-oil creams), creams are usually used for a variety of purposes including cleansing, emollient effects, and as a vehicle for drug substances such as local anesthetics, antiinflammatories (NSAIDs or corticosteroids), hormones, antibiotics, antifungals or counter-irritants.

[0083] 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- 1,3-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.

[0084] 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. In some embodiments, 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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, shea butter, 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.

[0089] 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.

[0090] The amount of fragrances in the topical composition is typically about 0.001% to about 0.5%.

[0091] Ointments are compositions in which oil and water are provided in a ratio of from 7 : 1 to 2: 1, from 5: 1 to 3: 1, or 4: 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.

[0092] Lotions arc low- to medium-viscosity topical preparations. Most lotions arc 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. In another embodiment, a berberine can be provided to the individual as a composition comprising emulsifying ointment BP, isopropyl myristate, hydroxyethylcellulose, glycerol, phenoxyethanol, propylene glycol and water.

[0093] In another embodiment, a berberine can be formulated with a delivery vehicle. For example, in a suitable embodiment, a composition of the invention may comprise liposomes in which at least some of the inhibitor of lactic acid fermentation, is incorporated.

[0094] By way of further example, suitable deliver}' 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 nanocmulsions, nanoparticlcs of various sizes, microparticles of various sizes, polystyrene microspheres of various sizes, titanium dioxide particles and solid lipid particles.

[0095] 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.

[0096] In a suitable embodiment, a composition of the invention may be formulated for use in combination with a device which enhances transdermal penetration of the composition. The composition may thus be applied to the area requiring troproelastin and / or elastin production. Merely by way of example such devices may include devices which produce electrical current such as galvanic devices, lasers, devices which produce vibrations such as ultrasound, and massage devices which enhance blood circulation around the target cells or target areas.

[0097] In some embodiments, the berberine may be delivered to the subject via transdermal delivery mechanism. Non-limiting examples of transdermal delivery mechanisms include microneedles or dissolvable microneedles, and the microneedles may be fabricated into a form of microneedle patch. In some embodiments, the composition may include one or more bioenhancers. Bioenhancers include any compound or composition that aids in the transport of another compound across epithelial membranes. Bioenhancers include P-glycoprotein inhibitors, compounds that reverse P-glycoprotein-mediated efflux, limit metabolism of active agents, increase gastric emptying time and intestinal motility, reduce degradation of the active agent by hydrochloric acid, modify cell membrane permeability, produce a cholagogue effect, modify the bioenergetics and thermogenic properties of the active agent, suppress first pass metabolism, and inhibit metabolizing enzymes, stimulate gamma glutamyl transpeptidase, enhance the uptake of amino acids, and the like and combinations thereof. Tn some embodiments, the bioenhancers may be herbal or neutraceutical bioenhancers. Examples of bioenhancers encompassed by the invention include pipeline, quercetin, gcnistcin, naringin, sinomcninc, glycyrrhizin, nitrile glycosides cuminum cyminum, zingiver officinale, lysergol, allium sativum, aloe vera, and the like and combinations thereof. In some embodiments, the biocnhanccrs may be liposomes, microspheres, nanoparticles, transferosomes, ethosomes, nanoemulsions, microemulsions, lipid based systems, polymeric micelle formulations, ketoprofen- loaded solid lipid nanoparticles, and the like, which can be made from beeswax, carnauba wax, or other natural waxes and solid lipids, and combinations thereof. In some embodiments, the bioenhancers may be liposomal enhancers such as, for example, ginkgo biloba lipid-based systems, silybin lipid-based systems, ginseng lipid-based systems, hawthorn lipid-based systems, quercetin lipid-based systems, curcumin lipid- based systems, and the like and combinations thereof. In further embodiments, the bioenhancers may be capsaicin transferosomes, colchicine tranferosomes, vincristine tranferosomes, and the like and combinations thereof, which may find particular use as natural skin penetration agents.

[0098] The amount of bioenhancer in the composition may be from about 0.05% to about 20% (w / w), relative to the total amount of the composition or in some embodiments, from about 0.1% to about 10% (w / w), relative to the total weight of the composition, from about 0.1% to about 5% (w / w), relative to the total amount of the composition, from about 0.1 % to about 2% (w / w), relative to the total amount of the composition, or any range or individual concentration encompassed by these example ranges. In some embodiments, the composition may further include an anti-inflammatory compound such as methotrexate, tofacitinib, 6-mercaptopurine, azathioprine sulphasalazine, mesalazine, olsalazine chloroquinine / hydroxychloroquinine, penicillamine, aurothiomalate (intramuscular and oral), azathioprine, cochicine, corticosteroids (oral, inhaled, and local injection), a beta-2 adrenoreceptor agonist (salbutamol, terbutaline, salmeteral), xanthine (theophylline, aminophylline), cromoglycate, nedocromil, ketotifen, ipratropium and oxitropium, cyclosporin, FK506, rapamycin, mycophenolate mofetil, leflunomide, an NSATD (e.g. ibuprofen), a corticosteroid (e. g. prednisolone), a phosphodiesterase inhibitor, an adensosine agonist, an antithrombotic agent, a complement inhibitor, an adrenergic agent, an agent that interferes with signalling by proinflammatory cytokines such as TNF or IL-1 (e.g., a NIK, IKK, p38 or MAP kinase inhibitor), an IL-1 converting enzyme inhibitor, a T-ccll signalling inhibitor (e.g. a kinase inhibitor), a metalloproteinase inhibitor, sulfasalazine, a 6-mcrcaptopurinc, an angiotensin converting enzyme inhibitor, a soluble cytokine receptor (e.g. soluble p55 or p75 TNF receptors and the derivatives p75TNFRigG (etanercept) and p55TNFRigG (Lenercept), siL-lRI, siL-lRII, siL-6R), an antiinflammatory cytokine (e.g. IL-4, IL-1 0, IL- 11 , IL- 13 and TGF), celecoxib, folic acid, hydroxychloroquine sulfate, rofecoxib, etanercept, infliximab, adalimumab, certolizumab, tocilizumab, abatacept, naproxen, valdecoxib, sulfasalazine, methylprednisolone, meloxicam, methylprednisolone acetate, gold sodium thiomalate, aspirin, triamcinolone acetonide, propoxyphene napsylate / apap, folate, nabumetone, diclofenac, piroxicam, etodolac, diclofenac sodium, oxaprozin, oxycodone HC1, hydrocodone bitartrate / apap, diclofenac sodium / misoprostol, fentanyl, anakinra, tramadol HC1, salsalate, sulindac, cyanocobalamin / fa / pyridoxine, acetaminophen, alendronate sodium, prednisolone, cortisone, betamethasone, morphine sulfate, lidocaine hydrochloride, indomethacin, glucosamine sulf / chondroitin, amitriptyline HCL sulfadiazine, oxycodone HCV acetaminophen, olopatadine HC1 misoprostol, naproxen sodium, omeprazole, cyclophosphamide, rituximab, IL-1 TRAP, MRA, CTLA4-IG, IL-18 BP, anti-IL-12, Anti-ILIS, BIRB-796, SCIO-469, VX-702, AMG- 548, VX-740, Roflumilast, IC-485, CDC-801, S1PI agonists (such as FTY720), a PKC family inhibitor (e.g. Ruboxistaurin or AEB-071 ) or Mesopram, budenoside; epidermal growth factor; a corticosteroid; cyclosporin, sulfasalazine; an aminosalicylate; 6- mercaptopurine; azathioprine; metronidazole; a lipoxygenase inhibitor; mesalamine; olsalazine; balsalazide; an antioxidant; a thromboxane inhibitor; an IL-1 receptor antagonist; an anti-IL-1 monoclonal antibody; an anti-IL-6 monoclonal antibody; a growth factor; an elastase inhibitor; a pyridinyl-imidazole compound; an antibody to or antagonist of other human cytokines or growth factors (e.g. TNF, LT, IL-1, IL-2, IL-6, IL-7, IL-8, IL-12, IL-15, IL-16, IL-23, EMAP-II, GM-CSF, FGF, and PDGF); a cell surface molecule (e.g. CD2, CD3, CD4, CD8, CD25, CD28, CD30, CD40, CD45, CD69, or CD90 or their ligands); methotrexate; cyclosporine; FK506; rapamycin; mycophenolate mofetil; leflunomide; an NSAID (e.g. ibuprofen); a corticosteroid (e.g. prednisolone); a phosphodiesterase inhibitor; an adenosine agonist; an antithrombotic agent; a complement inhibitor; an adrenergic agent; an agent that interferes with signalling by proinflammatory cytokines such as TNF 5 or IL-1 (e.g. a NIK, IKK, or MAP kinase inhibitor); an IL-1 converting enzyme inhibitor; a TNF converting enzyme inhibitor; a T-cell signalling inhibitor such as kinase inhibitors; a metalloproteinase inhibitor; sulfasalazine; azathioprine; a 6-mcrcaptopurinc; an angiotensin converting enzyme inhibitor; a soluble cytokine receptor (e.g. soluble p55 or p75 TNF receptors, siL-lRI, siL-lRII, siL-6R), an antiinflammatory cytokine (e.g. IL-4, IL-1 0, IL- 11, IL- 13 or TGF), therapeutic agents that target an intrinsic checkpoint blockade, such as, for example, the gene encoding Cytokine-inducible SH2-containing protein (CISH), antibody BGB-A317, Nivolumab, or Pembrolizumab, atezolizumab, avelumab, durvalumab, ipilimumab, and the like and combinations thereof. The amount of antiinflammatory in the composition may be from about 0.01% to about 5% (w / w), relative to the total amount of the composition, or in some embodiments, from about 0.1% to about 1% (w / w), relative to the total amount of the composition, or any range or individual concentration encompassed by these example ranges.

[0099] In some embodiments, the composition may further include an antiseptic compound. The antiseptic compound is not particularly limited, and in some embodiments may include, for example, iodine, manuka honey, octcnidinc dihydrochloridc, phenol, polyhexanide, sodium chloride, sodium hypochlorite, calcium hypochlorite, sodium bicarbonate, methyl paraben, and sodium dehydroacetate. The amount of the antiseptic in the compositions may be from about 0.01 % to about 5% (w / w), relative to the total amount of the composition, or in some embodiments, from about 0.1% to about 1%, relative to the total amount of the composition, or any range or individual concentration encompassed by these example ranges.

[0100] In some embodiments, the composition may further include an anti-acne compound. The anti-acne agent is not limited and includes, for example, salicylic acid, benzoyl peroxide, and the like and combinations thereof. The amount of the anti-acne compound in the compositions may be from about 0.01% to about 5% (w / w), relative to the total amount of the composition, or in some embodiments, from about 0.1% to about 1% (w / w), relative to the total amount of the composition, or any range or individual concentration encompassed by these example ranges.

[0101] In some embodiments, the composition may further include analgesic agent. The analgesic agent is not particularly limited and includes, for example, methyl salicylate, codeine, morphine, methadone, pethidine, buprenorphine, hydromorphine, levorphanol, oxycodone, fentanyl, a non-steroidal anti-inflammatory drug, and the like and combinations thereof. The amount of the analgesic agent in the compositions may be from about 0.01% to about 5% (w / w), relative to the total amount of the composition or in some embodiments, from about 0.1% to about 1% (w / w), relative to the total amount of the composition, or any range or individual concentration encompassed by these example ranges.

[0102] In some embodiments, the composition may further include depigmentation agent. The depigmentation agent is not particularly limited and includes, but are not limited to: hydroquinone, monobenzyl ether of hydroquinone, azelaic acid, kojic acid, mequinol, retinoids, soy proteins, alpha-hydroxy acids, trichloroacetic acid, salicylic acid, hydroquinone-beta-D-glucopyranoside, paper mulberry, glabridin, 4-isopropylcetchol, aleosin, N-acetyl-4-S-cycteaminylphenol, N-propionyl-4-S-cysteaminylphenol, N- acetyl glucosamine, tranexaminc acid, undecylenoyl phenylalanine, an alpha MSH antagonist, phytic acid or combinations thereof.

[0103] The berberines as referred to herein can be present in the composition in an amount ranging from 0.001 % to 10%, e.g. from 0.001 % to 5%, 0.005% to 5%, 0.01 % to 5%, 0.1% to 5%, 1% to 5%, 0.001% to 1%, 0.001% to 0.1%, 0.001 to 0.01%, or 0.001 to 0.005% of the total weight of the composition.

[0104] 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).

[0105] 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.

[0106] Throughout this specification and the statements 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.

[0107] 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.

[0108] 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.

[0109] 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. 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.

[0110] EXAMPLES

[0111] Methods

[0112] Human dermal fibroblasts were treated with 10 ng / ml TGF-01 or 1 pM BBR-HS. Seventy two hours later, cell culture supernatants were harvested and used to assay for monomeric elastin using a commercial ELTSA kit (see Figure 1). Error bars are ± SEM of 30 independent experiments, with two technical replicates each Statistical significance was assessed using one-way analysis of variance (ANOVA) followed by Tukey’s multiple-comparison test. ** p < 0.01, ****. p < 0.0001.

[0113] Human dermal fibroblasts were treated with the indicated concentration of berberine and their salt derivatives in Figure 2. Twenty-four hours later, cell culture supernatants were harvested and used to assay for elastin using a commercial ELISA kit (see Figure 2). Error bars are ± SEM of independent experiments.

[0114] Human dermal fibroblasts were treated with the indicated concentration of compounds in Figure 3. Seventy two hours later, cell culture supernatants were harvested and used to assay for tropoelastin using a commercial ELISA kit (see Figure 3). Error bars are ± SEM of three independent experiments.

[0115] Human dermal fibroblasts were treated with the indicated concentration of compounds in Figure 4. Seventy two hours later, cell culture supernatants were harvested and used to assay for tropoelastin using a commercial ELISA kit (see Figure 4). Error bars are ± SEM of three independent experiments.

Claims

CLAIMS1. A method of promoting tropoelastin synthesis in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

2. The method of claim 1, wherein the salt of berberine is berberine hydrogen sulphate, berberine hydrochloride or berberine sulfate.

3. The method of claims 1 or 2, wherein the derivative of berberine is a protoberberine alkaloid, optionally wherein the protoberberine alkaloid is epiberberine, palmatine, coptisine, jatrorrhizine or berberrubine.

4. The method of any one of claims 1 to 3, wherein the subject is a mammalian subject, such as a human subject.

5. The method of any one of claims 1 to 4, wherein the method comprises promoting elastin synthesis in dermal fibroblast of a skin tissue in the subject.

6. The method of any one of claims 1 to 5, wherein the method comprises promoting tropoelastin synthesis in dermal fibroblasts of a skin tissue in the subject.

7. The method of any one of claims 1 to 6, wherein berberine, or a salt, solvate, derivative or prodrug thereof is provided at a dose of less than 1 pM (or less than 0.33pg / ml).

8. The method of any one of claims 1 to 7, wherein the method a) improves the appearance of aging or sagging skin; b) improves skin firmness and / or elasticity or c) promoting a youthful appearance.

9. The method of claim 8, wherein the method is a non-therapeutic or cosmetic method.

10. The method of any one of claims 1 to 9, wherein the method comprises administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the skin of the subject.

11. The method of any one of claims 1 to 10, wherein the botanical extract is extracted from Berberis vulgaris, Phellodendron amurense, or Coptis chinensis.

12. The method of any one of claims 1 to 1 1 , wherein the method comprises topically and / or transdermally administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

13. The method of any one of claims 1 to 12, wherein the method further comprises administering retinol to the subject.

14. A method of promoting tropoelastin synthesis in a dermal fibroblast, the method comprising contacting the dermal fibroblast with berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine.

15. A method of improving the appearance of aging or sagging skin in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

16. A method of improving skin firmness and / or elasticity in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

17. A method of promoting a youthful appearance in a subject, the method comprising administering berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine, to the subject.

18. Berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine for use in promoting tropoelastin synthesis in a subject.

19. Use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for promoting tropoelastin synthesis in a subject.

20. Use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for treating a cutaneous wound.

21. The use of claim 20, wherein the medicament promotes de novo synthesis of tropoelastin within the wound site.

22. Use of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine in the manufacture of a medicament for treating a disease associated with a loss of tropoelastin and / or elastin fibres.

23. The use of claim 22, wherein the disease is lung emphysema, vascular aneurysm autosomal dominant cutis laxa (ADCL), supravalvar aortic stenosis (SVAS), Williams Beuren Syndrome (WBS) or mid-dermal elastolysis (MDE).

24. The use of claim 23, wherein the disease is UV-induced, such as photoaging.

25. A composition comprising berberine, or a salt, solvate, derivative or prodrug thereof, or comprising a botanical extract comprising berberine.

26. The composition of claim 25, wherein the composition comprises less than 0.0005% (w / v) of berberine, or a salt, solvate, derivative or prodrug thereof, or a botanical extract comprising berberine.

27. The composition of claims 25 or 26, wherein the composition is a topical composition.

28. The composition of any one of claims 25 to 27, wherein the composition is a cosmetic composition.

29. The composition of any one of claims 25 to 27, wherein the composition is a pharmaceutical composition.

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