WO2011019668A1 - Topical cosmeceutical formulation and methods of making and using same - Google Patents

Abstract

This invention relates to topical cosmeceuticals, and formulations thereof. In one embodiment, it relates to heat-enhanced transdermal delivery of cosmeceuticals using liposome-encapsulated delivery vehicles

Description

TOPICAL COSMECEUTICAL FORMULATION AND

METHODS OF MAKING AND USING SAME

CROSS-REFERENCE TO RELATED APPLICATIONS

[001] This application claims priority to United States Provisional Patent Application No. 61/232,465 entitled "Topical Cosmeceutical Formulation and Methods of Making and Using Same" and filed August 9, 2009. The contents of this United States

Provisional Patent Application are incorporated herein by reference in its entirety as if set forth verbatim.

FIELD

[002] This invention relates to topical cosmeceuticals, and formulations thereof. In one embodiment, it relates to heat-enhanced transdermal delivery of cosmeceuticals using liposome-encapsulated delivery vehicles.

BACKGROUND

[003] Cosmeceuticals (cosmetic products with biologically active components) are formulated to enhance the efficacy of cosmetics. In one instance, cosmeceutical formulations are administered topically to minimize or reverse the effects of aging on the appearance of skin. Complex cosmeceutical formulations are often formulated into a delivery system, such as particles, emulsions, liposomes, or the like.

[004] Liposomes can be used as delivery systems for pharmaceuticals and other topically administered materials. When liposomes are applied to the skin, they are able to flow into and through the stratum comeum, and may also be able to merge with cellular membranes and release their active materials within the cell. Thus, the liposomal delivery system is quite effective and efficient as most aqueous-based materials are not able to penetrate the skin's permeability barrier and reach the dermal layer.

[005] Therefore, new formulations and methods and systems for their administration that enhance their efficacy, including delivery vehicles and mechanisms, as well as combinations of cosmecutical ingredients are described herein. SUMMARY

[006] The foregoing needs are met, to a great extent, by the present disclosure, wherein in one of several aspects of the disclosure, a cosmeceutical composition for topical administration is provided, comprising: a yeast cell lysate; a fatty acid-peptide conjugate; a plant stem cell extract; an enzyme or coenzyme having antioxidant activity; and a microalgae cell extract.

[007] Another aspect includes the above formulation incorporated into a liposomal delivery system, and/or when the beneficial effects of the composition is enhanced when heated above room temperature (e.g. above 24⁰C).

[008] In yet another aspect of the disclosure, a method for enhancing the delivery of the above composition to skin is provided, comprising: topically applying the composition topically to the skin; and applying a controlled heat to the skin during or after administration.

[009] In another embodiment, the step of applying controlled heat may be achieved with an exothermically heated material. In various embodiments, the exothermically heated material may be in the form of a mitt, bootie, bonnet, mask or body strip.

[0010] In yet another aspect of the disclosure, an apparatus for heat- enhanced topical delivery of the above composition is provided, comprising: an amount of the composition of claim 1 applied to a skin-side of a self-heating fabricated material, wherein the fabricated material further comprises an exothermic material, wherein the exothermic material is activatable by an aqueous solution. In various embodiments of the present disclosure, the aqueous solution may be saline.

[0011] Other aspects of the invention are provided throughout the specification.

DETAILED DESCRIPTION

[0012] The following description relates generally to a topically administered

cosmeceutical formulation comprising at least five ingredients that are optimized for their synergistic effect on the appearance of skin. This cosmeceutϊcal formulation includes, as the active ingredients: yeast cell lysates, fatty acid-peptide conjugate, plant stem cell extract, bacterial antioxidant enzyme extract and niicroalgae cell extract.

[0013] It should be understood that the terms '"a" and "an^"' as used above and elsewhere herein refer to "one or more'^" of the enumerated components. It will be clear to one of ordinary skill in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms "a," "an" and "at least one" are used

interchangeably in this application.

[0014] Further, unless defined otherwise, all terms used herein have the same meaning as are commonly understood by one of skill in the art to which this invention belongs. AU patents, patent applications and publications referred to throughout the disclosure herein are incorporated by reference in their entirety.

Formulation

[0015] The active ingredients of the cosmeceutical formulation of the present invention are described in further detail below. a. Yeast Cell Lysates

[0016] Yeast cell lysates, which include live yeast cell derivatives ("LYCD"), are effective in the field of skin treatment and can be applied in the form of lotions, creams and oils. A wide variety of yeast cell lysates are contemplated to be effective in the various formulations of the present invention.

[0017] In one embodiment, the yeast cell lysate may be a stressed yeast cell lysate which is capable of enhancing oxygen respiration, promoting skin soothing and smoothing effects and simulating collagen synthesis. This stressed yeast cell lysate may contain tissue respiratory factor ("TRF"'), which is a live yeast cell derivative that has been shown to have therapeutic applications in skin cosmetics.

[0018] Live yeast cell derivatives can be manufactured by fermenting various types of yeast culture, such as Saccharomyces cerevisiae. One method of producing a stressed yeast cell lysate is by subjecting yeast cells to irradiation with ultraviolet light and heat, which causes the yeast cells to produce a variety of protective substances. Once this has occurred, the yeast cell walls are broken down through use of a proteolytic enzyme (i.e., they are lysed), and then centrifuged to leave only cellular protoplasm behind.

[0019] TRF is composed of the low molecular weight glycosidic : peptide fractions of the lysate, with a ratio of 1 :3. The residual glycopeptide linkages are through the amino acid asparagine residues. Because TRF is prepared from cell lysates of live yeast cells, additional trace quantities of coenzymes, vitamins, amino acids and minerals, which are also included in the lysate, are co-isolated, which enhances the therapeutic capabilities of TRF in these pharmaceutical/cosmetic preparations. TRF's moisturizing effect is accomplished by increasing uptake of moisture by nascent protein and increasing oxygen utilization in the skin.

[0020] For example, it has been shown that TRF can increase oxygen respiration in cultured human dermal fibroblasts by up to 1 17% in comparison to an untreated control. Further, it has also been demonstrated that TRF can promote procollagen synthesis and significantly increase hyaluronic acid synthesis in human dermal fibroblasts when compared to an untreated control.

[0021] One particularly effective TRF is commercially available as Biodynes-TRF and is manufactured by Brooks Industries (South Plainfield, NJ.). b. Fatty Acid-Peptide Conjugates

[0022] Formulations of fatty acid-peptide conjugates in the form of biomimetic oligopeptides used in the various formulations of the present invention may be natural or synthetic oligopeptides having a sequence of 20 amino acids or less, and sometimes 3, 4 and 5 amino acids, which correspond to a fragment of a protein of the extracellular matrix in the dermis, such as al-pro-collagen, a2-collagen 1 , elastin, tropoelastin, fibronectin, laminin-5 or immunoglobulin IgG.

[0023] Biomimetic oligopeptides, upon application, may act as messengers of a fake aggression on skin cells, thereby triggering activation of a mechanism similar to wound healing or matrix renewal. They have been found to be able to exercise feedback control on the process of connective tissue renewal and cell proliferation, for example by stimulating neo-synthesis of collagen I, III, IV, fibronectin, Iaminin-5 and/or

glycosaminoglucans by fibroblasts.

[0024] Non-limiting examples of biomimetic oligopeptides used in the compositions of the present invention may include the tripeptide Palmitoyl-GIycyl-Histidyl-Lysine (PaI- GHK), the tetrapeptide Palmitoyl-Glycyl-Glutaminyl-Prolyl- Arginine (PaI-GQPR) and a mixture of PaI-GHK and PaI-GQPR, also known as Matrixyl™ 3000 which is manufactured by Sederma S.A.S. (Le Perray-en Yvelines Cedex, France). c. Plant Stem Cell Extracts

[0025] A number of plant cells have been used in cosmetic formulations for the treatment of various skin conditions. For example, dedifferentiated plant cells, or piant "stem cells" have been studied for their therapeutic applications for treating skin. One such plant, known as Uttwiler spatlauber, is a variety of a Swiss apple that has been studied for its excellent storability properties.

[0026] Plant stem cells have a complex matrix of constituents made up of salts, acids, polyphenols, sugars, fats, proteins and other components. In addition to known components, there is also an unknown fraction of components which may have cosmetic applications. Specifically, it has been shown that plant stem cells contain specific epigenetic factors whose function is to maintain the self-renewal capacity of stem cells.

[0027] Plant cell extracts have been used for many years in skin care formulations. For instance, it is known to solubilize plant cells in suspension cultures and extract the oil and water soluble agents into empty liposomes to improve both the stability of the extracted agents and their transportation into skin.

[0028] One method of extracting plant stem cells involves triggering a plant's wound healing mechanism in order to induce the formation of callus cells. This wound healing tissue consists of stem cells. The callus cells, including stem cells, can then be harvested and cultivated in a suspension and homogenized together with liposomes to encapsulate and stabilize both oil and water soluble components. One such process, PhytoCellTec™ (Mibelle AG Biochemistry, Switzerland) involves an eight step procedure for preparing plant stem cells: 1) selecting/taking away a small piece of the plan; 2) wounding of plant material to induce callus formation; 3) incubation on agar plates; 4) harvesting of developed callus; 5) cultivation until complete dedifferentiation to obtain stem cells; 6) transfer of the stem cells into a suspension (liquid media); 7) disruption of the stem cells wall; and 8) encapsulation of the stem cell content into liposomes.

[0029] At least one plant stem cell extract is present in the formulations of the present invention. A non-limiting example of a particularly effective plant stem cell is supplied as PhytoCellTec™ from Malm domestica, manufactured by Mibelle AG Biochemistry (Switzerland). d. Bacterial Antioxidant Enzyme Extracts

[0030] The formulations of the present invention also further comprise a safe and effective amount of one or more enzymes, enzyme inhibitors or enzyme activators (coenzymes). Examples of enzymes are lipases, proteases, catalase,

superoxidedismutase, amylases, glucuronidases, peroxidases, in particular glutathione peroxidase or lactoperoxidase, ceramidases, hyaluronidases. All of these enzymes may be obtained by extraction from bacteria using fermentation processes. In one

embodiment, the enzyme is an anti-oxidant selected from the group of superoxide dismutase, catalase and peroxidase. Furthermore, also contemplated in various formualations of the present invention are enzymes and enzyme complexes that exhibit superoxide dismutase-like properties, glutathione peroxidase-like activity and catalase- like activity.

[0031] In one exemplary embodiment, the enzyme or coenzyme activity is involved in the protection against oxidation and is enhanced by applying effective amounts of heat. A number of skin treatment formulations may exhibit a change in the rate of activity due to the presence of heat. In some formulations, the heat- activated ingredient becomes active or exhibits an increased level of enzymatic activity when it is heated to a temperature range of between 40° C and 60° C. For example, various enzyme complexes exhibit higher superoxide dismutase-like activity proportional to increases in heat.

Alternative enzyme complexes can exhibit higher glutathione peroxi date- like activity or catalase-like activity as well in response to higher temperatures.

[0032] In addition, various enzyme complexes can also exhibit greater stability of enzymatic activity in response to higher temperatures. Further, some enzyme complexes may also exhibit higher levels of enzymatic activity or stability in response to an increase in UVA irradiation as well. One non-limiting example of a particularly effective heat enhanced ingredient is found in an extract of Thermus thennophilus, which was discovered in a deep-sea hydrothermal vent in Guaymas Basin (Gulf of California at 2000 m depth). The fermentation of Thermus thennophilus for producing proteins is disclosed in WO 02/066668 A2, incorporated herein by reference. The use of extracts from Thermus thermophilus cultures in cosmetic compositions is known to modulate the cutaneous concentration of ceramides, to stimulate the immune system to provide protection as a detoxifying agent and against free radicals, especially oxygen peroxide.

[0033] Specifically, various Thermus thermophilus extracts have been shown to exhibit anti-oxidant properties such as removal of superoxide anions and peroxidase activity, as well as an increase in resistance to UVA-induced peroxidation. Thermus thermophilus extracts have also been shown to confer upon UVA- irradiated fibroblasts the ability to retain anchor collagen at a high rate and retain high levels of catalase activity. Various other benefits may include reduced membrane lipid peroxidation and reduced UVA- induced oxidative lesions of DNA in fibroblasts. A Thermus thermophilus extract is available commercially from Sederma, (Le Perray-en Yvelines Cedex, France) under the product label Venuceane™. e. Microalgae Extracts

[0034] A safe and effective amount of a microalgae-derived active ingredient is also included in the various foπnulations of the present invention. Various microalgae can be found in inhospitable environments such as salt lakes and undergo a variety of stresses such as high osmotic pressure, high UV-levels and high oxygen levels. Thus, because they are capable of withstanding inhospitable environments, various microalgae extracts have been shown to confer beneficial effects in cosmeceutical applications. Microalgae are a rich natural source of vitamins, minerals and other components essential for skin care. In particular, carotenoids are an abundant component of microalgae extracts.

[0035] Although a number of compounds derived from algae may be utilized in the present invention, an active ingredient derived from Dunaliella salina is particularly effective in the treatment of skin. Extracts of Dunaliella salina have been shown to stimulate ATP-production of mitochondria and improve cell turnover in skin cells. A commercially optimized form of Dunaliella salina is available from Pentapharm (Basel, Switzerland) under the trade name PEPHA^®-CTIVE.

[0036] The ingredients described above can be mixed in an aqueous solution at varying amounts wherein the concentrations of each active ingredient can be optimized using routine optimization. For example, the concentration of ingredients may be prepared as follows:

[0037] The various formulations may also optionally include a wide range of excipients. Some suitable excipients commonly used in the cosmetic and personal care industry are described in The CTFA Cosmetic Ingredient Handbook, (9^l Ed., 2002), which is incorporated by reference herein. These ingredients will be used in amounts which are conventional.

Liposomes

[0038] Certain cosmetic formulations contain ingredients that must be capable of penetrating through various external layers of the skin to interact with cells blow these external layers. This sometimes requires the use of complex delivery systems to make otherwise unpenetrating and unstable compounds more efficacious. In one aspect, ingredients that are highly charged, insoluble and/or have a high molecular weight sometimes have limited usefulness unless they are coupled to a carrier or incorporated into a delivery vehicle.

[0039] The problems associated with successful administration of cosmetic formulations are compounded by the fact that certain types of compounds cannot exhibit their effects unless they are able to penetrate the skin and contact cells beneath the skin's external layers. For example, the use of compounds depends on their ability to interact with cellular components at or near the dermis level. However, such compounds exhibit poor penetrability when delivered in aqueous solution alone.

[0040] Many different approaches have been suggested for enhancing skin absorption of cosmetic formulations. Delivery vehicles such as liposomes have been described for use in the cosmetic field. An advantage of using liposome formulations is the ability of these substances to mimic naturally occurring cellular membrane substituents. This encourages absoiption and intracellular uptake, which results in delivery of the liposome contents into the inner skin layers, and cell cytoplasm.

[0041] Liposomes are tiny spherical lipid vesicles which form when phospholipids hydrate in water. These vesicles' membranes are similar in structure to the outer membranes of biological cells. Phospholipids are natural moisture barrier components of the skin, aiding in cellular repairs and turnover. Under low shear mixing conditions, they tend to be multilamellar structures comprised of oriented bilayers of phospholipids molecules surrounding concentric aqueous compartment. Further processing can produce unilamellar liposomes, which are small in size, uniform, and stable

[0042] One can think of a liposome as a hollow, empty, cell-like membrane. Liposomes are generally produced using soy lecithin, whose main compound is phosphatidylcholine. Lecithin processed through a microfluidizer forms the unilamellar structure mentioned earlier. Because phosphatidylcholine is known as a penetration enhancer, this property is usually associated with liposomes. Liposomes are the vesicles which transport cosmetic agents more easily into the horny layer; moreover, the conditioning effect causes the horny layer to become a depot for these agents. Measurements of systematically active pharmaceuticals/cosmeceuticals reveal that an increase of penetration is not necessarily correlative with an increase of permeation. Actually, permeation of active agents is often slowed by phosphatidylcholine in such a way that a high permeation peck in the beginning of the application is prevented. Instead, a more continuous permeation takes material out of the horny layer depot into the living part of the body over a longer period of time. This property makes the incorporation of phosphatidylcholine and liposomes very attractive or the application of vitamins, provitamins and other substances influencing the regenerating ability of the living epidermis.

[0043] Liposomes can generally be made available in two foπns, dispersed and emulsified. Dispersed liposomes are produced with solutions of water-soluble agents which are captured in the interior compartments of the vesicle, and are present in the continuous space of the dispersion. Dispersed liposomes are generally 200nm or less in diameter, and are translucent in appearance. Emulsified liposomes capture lipids within the bilayer of the vesicle, which tends to enlarge the size of the vesicle. Emulsified liposomes range in size up to 300nm, and are opaque in appearance.

[0044] The capture capability of one gram of phospholipids-based liposome with an average diameter of 70nm is 2.75 ml/gm of aqueous solution in a dispersed liposome. The lipid capture capacity in an emulsified liposome is 1.67 ml/gm. Spherical liposomes of this size may contain an excess of 30,000 phospholipid molecules with an aggregate "Molecular Weight" exceeding 24,000,000. Regardless of capture capability, all cosmetic liposmes contain "free" material in the continuous phase.

[0045] Some of the earliest and most effective liposomes for skin care cosmetics were primarily composed of phospholipid choline esters of phosphoric acid and a mixture of fatty acid diglycerid.es. Phosphatides are constituents of living cells and affect cell permeability, metabolism, and body function. In liposome forms, cosmetic ingredients exhibit better stability, penetration, and efficacy at lower usage levels.

[0046] The general hypothesis for the mechanism of action of liposomes is understood as follows:

[0047] 1. When liposomes are "filled" with various actives, they can penetrate the stratum corneum and bring hydration and actives through the top layers of the epidermis, with penetration stopping at the "end" of the living epidermal cells.

[0048] 2. Liposomes are hydrated bilayer structures, which are readily compatible with the peridermal lipids (the skin's primary moisture barrier), which exist as lamellar layers.

[0049] 3. When liposomes are applied to the skin, they interact with proteins, carbohydrates and the lipids that compose cell membranes, in a manner similar to that of the cell membranes' own lipid bilayer. It is believed that liposomes partially bind to keratin in the homy layer of the skin.

[0050] A special quality of liposomes is that they enable water-soluble and water- insoluble materials to be used together in a formulation without the use of surfactants or emulsifiers. Water-soluble materials are dissolved in the water in which the

phospholipids are hydrated, and when the liposomes form, these materials are trapped in the aqueous center. The liposome wall, being a phospholipid membrane, holds fat- soluble materials, such as oil.

[0051] The following are various exemplary benefits of liposomes:

1. Reinforce the barrier function of the skin and reduce trans- epidermal water loss, thus providing an ideal moisturizing effect. 2. Can bring difficult-to-penetrate materials into the epidermis.

3. Pleasantly smooth feeling on skin.

4. Polyunsaturated vegetable phospholipids such as lecithin may enhance skin cell metabolism by making the membranes more fluid.

5. Phospholipids, particularly polyunsaturated phospholipids, may have an additional benefit: the liposomes ultimately reach the sebaceous glands, where the linoleic acid in these compounds can supplement the supply of efa (essential fatty acid). These glands require linoleic acid to function, and an insufficient supply can lead to increased pimple and blackhead formation.

6. Due to the similarity between liposomes and the membranes of skin cells, the releasing of materials captured within the liposome walls when in contact with skin cell membranes may be more than a coincidence.

7. Controlled delivery system

8. Biodegradable and non-toxic

9. Carry either/both oil and water payloads

10. Protein stabilization

11. Controlled hydration

12. Anti-acne properties

13. Liposomes have excellent substantivity to keratin (skin and hair) and moisturizers encapsulated in liposomes have been shown to be resistant to washing and removal from the skin.

[0052] Liposomal preparations may be dosage dependant, requiring a good level of high- quality liposomes within a given formulation, In addition to the dosage dependence for efficacy, the following principles are important in topical liposome formulations: [0053] Surfactants significantly affect the behavior of liposomes. Soaps rapidly (albeit reversibly) break liposomes down into smaller particles; thus, liposomal liquid soaps or shampoos generally cannot easily be formulated. However, liposomes are quite stable in products with amphiphilic surfactants, such as ethanol or ethylene glycol (the vesicles then to be relatively small in alcohol-containing products.)

[0054] Liposomes which are most useful for the delivery of cosmetic ingredients are often complex formulations containing mixtures of different lipophilic substituents. These complex mixtures allow for optimization of the physical properties of the liposomes, such as pH sensitivity, temperature sensitivity and size. The inventors have recognized that certain temperature sensitive liposome constituents can be used to formulate cosmetic ingredient-containing liposomes that destabilize at increased temperature. This has been shown to promote delivery of the liposome contents after application.

[0055] The inclusion of sterols in liposomes is also possible. In general, the presence of sterols in liposome formulations results in enhanced stability, both in vitro and in vivo. Liposome formulations for the delivery of biomolecules which contain organic acid derivatives of sterols, such as cholesterol or vitamin D, have been reported to be easier to formulate than their non-derivatized water-insoluble equivalents

[0056] Lipid aggregates can take the form of completely closed structures made up of a lipid bilayer containing an encapsulated aqueous volume (i.e. unilamellar liposomes), or they may contain more than one concentric lipid bilayer separated by an aqueous volume (i.e. multilamellar liposomes). Each lipid bilayer is composed of two lipid monolayers, each of which has a hydrophobic (nonpolar) "tail" region and a hydrophilic (polar) "head" region. In the bilayer, the hydrophobic "tails" of the lipid monolayers orient toward the inside of the bilayer, while the hydrophilic "heads" orient toward the outside of the bilayer. It is within the aqueous phase that the biomolecules become entrapped in the liposome, unless the biomolecule is in the form of a lipid-biomolecule conjugate, in which case the biomolecule may become embedded within the bilayer. [0057] Liposomes may be made by a variety of techniques known in the art. (See, for example, Bangham et al ., J. MoI. Biol., 13: 238-252(1965), incorporated by reference in its entirety herein). These methods generally involve first dissolving and mixing the lipids in an organic solvent, followed by evaporation. Then an appropriate amount of the aqueous phase is mixed with the lipid phase, and then allowed to incubate for a sufficient time for the liposome to form. The aqueous phase will generally consist of the biomolecules in suspension with other solutes, such as buffers or sugars.

[0058] In addition to the biomolecule to be encapsulated, the liposomes of the present invention are comprised of the lipids, such as an anionic lipid (for example,

phosphatidylglycerol, phosphaticid acid or a similar anionic phospholipid analog), or a neutral lipid (for example, phosphatidylcholine or phosphatidylethanolamine). The present liposomal formulations may further include a lysolipid, such as

lysophosphatidylcholine, lysophosphatidylethanol amine, or a lyso form of a cationic lipid species.

[0059] The liposome can also include optional substituents, such as sterols, glycolipids, tissue or organ targeting substances such as antibodies or proteins, fatty acids, or any other natural or synthetic lipophilic or amph philic compounds.

[0060] Suitable sterols for inclusion into the liposome include, but are not limited to, cholesterol and Vitamin D, and are included in the liposome formulations as stabilizers.

[0061] An important aspect is the discovery that the efficacy of the liposome

formulations are enhanced by the presence of temperature sensitive substituents. See, for example, U.S. Patent No. 6,200,598, incorporated by reference in its entirety herein. Accordingly, the liposomes of the present invention can be designed to be sensitive to alterations in the temperature of the surrounding environment. The temperature- sensitivity of such liposomes allows the release of compounds entrapped within the interior aqueous space of the liposome, and/or the release of compounds associated with the lipid bilayer, at a target site that is either heated or that is at an intrinsically higher temperature than the site of application, which would be expected for application to the skin, since the inner skin layers are at a higher temperature than the external skin surface. [0062] Liposome bilayers of the present invention may include (in addition to a primary or main lipid component) lysolipid, or another surface active agent(s). The inclusion of lysolipid or another surface active agent in the liposome bϋayer enhances the release of compounds when the liposome temperature reaches the gel-to-liquid crystalline phase transition temperature of the primary lipid component. The presence of the lysolipid (or other surface active agent) also causes the liposome to release its contents at a slightly lower temperature than that achieved with liposomes composed solely of phospholipids. Liposomes of the present invention are particularly useful in skin delivery, where the liposome contains a compound to be delivered to the inner skin layers. The skin is either artificially heated above body temperature so that it is at or above the gel-to-liquid crystalline phase transition temperature, or the liposome is formulated such that the gel- to-liquid crystalline phase transition temperature of the liposome is at or near body temperature,

[0063] When liposomes are incubated for several minutes at temperatures in the region of the gel-to-liquid crystalline phase transition temperature (Tc) of the primary lipid composing the liposome, the liposome bilayer becomes permeable and releases solutes entrapped within the liposome into the surrounding environment. Such thermally- sensitive liposomes have been proposed. See, e.g., Yatvin et al., Science 202:1290 (1978), incoiporated by reference in its entirety herein.

[0064] U.S. Patent No. 5,094,854 (Ogawa et al.), incoiporated by reference in its entirety herein, discloses liposomes in which the osmotic pressure of the cosmetic component- containing solution entrapped in liposomes is 1.2-2.5 times higher than body temperature. The temperature range in which the liposome membrane becomes permeable to material release is stated to be in the range of 40-45 degrees centigrade.

[0065] Liposomes can comprise a lipid possessing a gel-to-iiquid crystalline transition temperature in the hyperthermic range (e.g., the range of from approximately 38-45 degrees centigrade). In one embodiment, phospholipids with a phase-transition temperature of from about 38-45 degrees centigrade are utilized. Such phospholipids include those whose acyl groups are saturated. A particularly preferred phospholipid is dipalmitoylphosphatidylcholine (DPPC). DPPC is a common saturated chain (C 16) phospholipid with a bilayer transition of 41.5 degrees centigrade. (Blume, Biochemistry 22:5436 (1983); Albon and Sturtevant, Proc. Natl. Acad. Sci. USA 75:2258 (1978)). Thermosensitive liposomes containing DPPC and other lipids that have a similar or higher transition temperature, and that mix ideally with DPPC (such 1,2-Dipalmitoyl-sn- Glycero-3-[Phospho-rac-(l -glycerol)] (DPPG) (Tc=41.5 degrees centigrade) and 1,2- Distearoyl-sn-GlyceiO-3-Phosphocholine (DSPC) (Tc=55.1. degrees centigrade.)) have been studied. Kastumi Iga et al, Intl. J. Pharmaceutics, 57:241 (1989); Bassett et al, J. Urology, 135:612 (1985); Gaber et al, Pharmacol. Res. 12:1407 (1995). Thermosensitive liposomes containing DPPC and cholesterol have also been described. Demel and De Kruyff, Biochim. Biophys. Acta. 457:109 (1976). Each of the above documents are individually incoiporated by reference herein.

|0066] The "primary lipid" in a liposome bilayer can be the main lipid component of liposome bilayer material. Thus, in a liposome bilayer composed of 70 mole % phospholipid and 30 mole % lysolipid, the phospholipid can be the primary lipid.

[0067] Liposomes suitable for the present invention can incorporate a relatively-water soluble surface active agent, such as a lysolipid, into a bilayer composed primarily of a relatively water-insoluble molecule, such as a di-chain phospholipid (e.g., DPPC).

Incorporation of the surface active agent in the gel phase of the primary lipid component enhances the release of contents from the resulting liposome when heated to the gel- liquid crystalline phase transition temperature of the primary lipid. Effective surface active agents are lysolipids, and a particularly effective surface active agent is monopalmitoylphosphatidylcholine (MPPC), Suitable surface-active agents are those that are compatible with the primary lipid of the bilayer, and that desorb when the lipid melts to the liquid phase. Additional suitable surface-active agents for use in

phospholipid bilayers include palmitoyl alcohols, stearoyl alcohols, palmitoyl, stearoyl, polyethylene glycol, glyceryl monopalmitate, glyceryl monooleate, ceramides, PEG- ceramides, and therapeutic lipids. Therapeutic lipids include, for example, C- 18 ether linked lysophoshpatidylchohline. [0068] In another embodiment, the liposomes incoiporate a bilayer-compatible lysolipid in a phospholipid bilayer, the lysolipid present in the bilayer at a concentration sufficient to enhance the release of contents (e.g., active agent or therapeutic agent) from the liposome, compared to the release of contents that would be achieved using a liposome composed of only lipid alone (i.e., without lysolipid). The contents may be contained within the interior of the liposome or in the liposome membrane. By "enhanced release", it is meant that either (a) that a greater percentage of contents is released at a given temperature, compared to the amount of contents released at that temperature by a lipsome with a bilayer composed of phospholipid only; or (b) the contents entrapped in the interior of the liposome are released at a lower temperature than the temperature at which release of contents would occur using a lipsome with a bilayer composed of phospholipid only.

[0069] Manufacturing liposomes may involve preparation of gel phase liposomes containing a phospholipid, an appropriate surface active agent (such as lysolipid), and the cosmetic ingredients to be encapsulated. The composition contains a percentage of lysolipid such that the surface active agent does not destabilize the membrane at processing temperatures where the bilayer is in the liquid phase, nor at the surface temperature of the skin where the bilayer is in the gel phase. Optimization of percentage of liposome constituents can easily be accomplished by routine optimization. The bilayer becomes unstable and permeable at temperatures at or just above normal human body temperature, resulting in release of entrapped material from the liposome interior.

[0070] Liposomes may be prepared by any of a variety of techniques that are known in the art. See, e.g., U.S. Pat. No. 4,235,871; Published PCT applications WO 96/14057; New RRC, Liposomes: A practical approach, IRL Press, Oxford (1990), pages 33-104; Lasic D D, Liposomes from physics to applications, Elsevier Science Publishers, Amsterdam, 1993; Liposomes, Marcel Dekker, Inc., New York (1983). Each of the above documents are incorporated by reference herein.

[0071] Generally, the method of preparing liposomes involves mixing the bilayer components in the appropriate proportions in an organic solvent, which is thereafter evaporated to form a dried lipid film. The film is rehydrated (at temperatures above the phase transition temperature of the lipid mixture) using an aqueous solution containing an equilibrating amount of the surface active agent and the desired ingredients to be encapsulated. The liposomes formed after rehydration can be extruded to form liposomes of a desired size.

Other Delivery Vehicles

[0072] Topical delivery systems are broadly categorized as either transdermal or dermal. Transdermal delivery (TDD) is the controlled release of cosmetic formulation components through intact skin to obtain beneficial levels systematically and to affect specified targets for specific puiposes such as anti-aging, among others. Dermal ingredient delivery (DDD) is similar to TDD except that the specified target is the skin itself. Cosmetic ingredients, including those for antiaging, known as cosmeceuticals, presumably may have the same site of action. The cosmetic and/or transdermal route is indeed desirable by the industry but the success of DDD and cosmetic active ingredient delivery (CAID) technologies remains limited and faces many challenges, one of which is low skin permeability that hinders the development of DDD/CA1D for

macromolecules.

[0073] Overcoming the skin barrier safely and reversibly while enabling percutaneous absorption is a fundamental problem in the field of CAID, TDD and DDD. Advances have been made in recent decades to overcome skin barrier properties, including physical means such as iontophoresis, sonophoresis and microneedles; chemical means using penetration enhancers; and biochemical means such as enzyme inhibition.

[0074] Accordingly, the formulations of the present invention may be delivered in vehicles other than liposomes, either dermally or transdermal Iy. Such delivery vehicles are well characterized in the cosmetic arts. For example, see Rosen, Meyer R. Delivery System Handbook for Personal Care and Cosmetic Products - Technology, Applications and Formulations, William Andrew Publishing, the contents of which are incorporated by reference herein. [0075] By way of non- limiting example, common skin penetration enhancement techniques include electrically assisted techniquies (ionotophoresis, electroporation, sonophoresis and photophoresis); Stratum comeum bypass (microneedles, stratum corneum ablation by, for example, chemical peels, microdermabrasion, microscission, laser, etc); non-liposome vesicles and particles (nanoparticles, high velocity particles such as powder injection, etc); and stratum comeum modification (hydration, chemical penetration enhancers.)

[0076] Of course, other skin penetration techniques may be contemplated to be within the scope and spirit of the current disclosure.

Heating Mechanisms

[0077] In one embodiment, the cosmeceutical formulations of the present invention are administered to the skin's surface in conjunction with the administration of heat. As described above, heat is known to enhance the efficacy of transdermal delivery and activity of various topically administrated cosmeceutical ingredients by increasing skin permeability and rate-limiting membrane permeability, as well as by directly enhancing enzyme activity. Diffusion through the skin is a temperature-dependent process, so raising the skin temperature will add thermodynamic drive. Heat is known to increase the activity of many of the active ingredients of the topical formulation. In addition, it confers biophysical affects on the proteins, lipids, and carbohydrates in the cell membranes to which the topical formulation is placed. Heating prior to or during topical application of a substance will dilate penetration pathways in the skin, increase kinetic energy and the movement of compounds in the treated area, thus facilitating absorption. Heating the skin after the topical application of a material will increase that material's absoiption. By increasing skin permeability and rate-limiting membrane permeability, increases in localized temperature can be seen which greatly enhance permeation through the skin. a. Chemically Induced Heating

[0078] One suitable mechanism for generating heat is through the use of an exothermic composition that generates heat upon activation by a secondary, readily available activator substance. Examples of exothermic compositions that can be used may come from the combination of water with strong acids, combining alkalis and acids, polymerization, thermite reactions, aluminum-based reactions, magnesium-iron-based reactions, anhydride reactions, and so forth. One particularly suitable, non-toxic exothermic composition is Lava Gel^{1 M} (manufactured by Forever Young International, Inc., Escondido, California, USA) which is known to exhibit a very controlled, regulated temperature for an extended period of time, with simply the addition of water or an electrolyte solution, such as saline water (as the secondary compound). However, other exothermic compositions may be used, accordingly to design preference, including compositions that require activation or moderation by more than one secondary compound or element.

[0079] By use of a non-electrical or non-fossil fuel heating source, the mechanisms for application as described herein can be considered as self-contained units, portable, and also, in some instances, disposable with minimal to no environmental consequence. With a regulated, controlled exothermic reaction, overheating can be avoided, as well as burns that occur from such overheating. The disposable embodiments can be of limited or of single use, whereby complications arising from reuse can be obviated. Thus, with respect to sanitation aspects, a limited use or single use unit minimizes the occurrence of cross- contamination, as commonly seen in conventional application systems. Also, with limited or single use products, they are of smaller size than institutional products.

Therefore, the exemplary embodiments for application can also be easily shipped, easily stored (e.g., suitcase, handbag, etc.), and much more affordable for the individual user.

Administration

[0080] The formulations described above can be administered by any means that other topical formulations can be administered. For formulations dependent on heat efficacy, they can be applied to the skin prior to application of a heat-generating appliance (mechanism), or can be applied directly to, embedded in (wholly or in-part), and so forth, into the heat-generating appliance (mechanism). Thus, various combinations of modes for application and modifications thereto (prior to, during, after, partially, to the skin, to the appliance, and so forth) are considered within the scope of this disclosure. [0081] In various embodiments, the heat-inducing appliance (mechanism) may be, as non-limiting examples, in the form of self-heating booties, mitts, masks, body strips, bonnets, and so forth. Descriptions of these appliances are provided below.

[0082] A self-heating mitt or bootie can be devised by a multi-layered material (i.e. a "fabricated material") containing an exothermic material, bounded by permeable and/or non-permeable/semi-permeable layers. For example, an outer layer can be formed from an open-ended enclosure with a front and a back covering with a seal about its perimeter, or the outer layer may be of a single and unitary construction. The open-ended enclosure can be shaped in the form of a mitt or glove capable of receiving a hand, or a foot or other extremity of the body. A heating layer can be singly configured or symmetrically configured to form layers of heating panels. For example, the heating layer can be comprised of a first outer fabric layer, an exothermic layer, and a second outer fabric layer. A nonwoven fabric or polymeric material can be utilized.

[0083] A liner can be placed interior to the heating layer and may be of a water- impermeable material to protect a hand or foot from direct exposure to the heating layer. Non-limiting examples of water-impermeable materials include natural and synthetic rubbers, polyvinyl chloride, ethylene- vinyl acetate copolymers, polyurethanes, acrylic ester polymers, polyamides and the like. The liner can be provided with a topical having heat-enhancible or heat- activated skin properties as described above. The liner may also include therapeutic compounds for pain relief such as camphor, menthol, capsaicin and witch hazel. Herbal products such as natural extracts like green tea, grapefruit seeds and yucca root may also be suitable for use as a topical as well.

[0084] The liner may be "pre-configured" with the formulations or "post-configured" with the formulations. In some embodiments, a pre-configured mitt/bootie, may require the removal of a seal or exposure of the formulation by some physical action, such as breaking a seal, shaking, and so forth. The seal may be waterproof, if so desired. Also the removal of the seal may occur after the mitt/bootie is exposed to the activating solution. [0085] Based on the above-described mitt/bootie, etc., having the exothermic material, a convenient mechanism for generating heat is presented, as well as a mechanism for applying the above-described topicals/compositions.

[0086] A self-heating mask can be similarly devised using the multi-layered material described above. In one of various self-heating mask embodiments, a temperature controlled facial mask with area-specific treatments can include a substantially planar mask body which is formed with cut-outs for a person's eyes, nose, mouth, etc. On the front side of the mask body, the face mask may include a number of treatment zones. The treatment zones can be coated with skin treatments/topicals, in the various manners, as described above. On the backside of the mask body, a self-heating or cooling substrate may be applied to provide temperature control to the face mask.

[0087] In some cases, it may be desirable to provide a mask for only treating the areas around the eyes, and in that case, the entire mask might resemble an eye mask.

Alternatively, in certain treatment procedures, small masks may be made having unique shapes to contour match a particular body part, such as a neck or shoulder for area- specific treatments.

[0088] Alternative embodiments of the mask may include multiple pouches having heating and/or cooling properties. More specifically, a substrate can include a backing panel such as polyethylene or an absoiptive non-woven. An upper panel, such as an absorptive non-woven, can be positioned over the backing panel and a plurality of seals are made to form a number of pouches. Each pouch contains an material, such as for heating (or cooling), or may be empty to provide neither heating (or cooling).

[0089] The pouches can also operate as treatment area to provide a number of very area- specific treatment zones. Accordingly, the mask may be made with any number of treatment zones, and any combination of heating, cooling or ambient (no heating or cooling) zones. In some instances, cooling is beneficial to treat Rosacea.

[0090] Straps may be formed about the mask to tie the mask onto the wearer's face to apply pressure to the mask and increase the contact area on the face. In some embodiments, the cosmeceutical formulations of the present invention can be distributed in the mask.

[0091] Accordingly, different conditions/locations can be selectively treated, if so desired. With a self-heated mask, selective treatment using any one or more of the formulations described above can be realized. The mask may be "pre-con figured" with the formulations or "post-configured" with the formulations. In some embodiments, a pre-configured mask may require the removal of a seal or exposure of the formulation by some physical action, such as breaking a seal, shaking, and so forth. The seal may be wateiproof, if so desired. Also the removal of the seal may occur after the mask is exposed to the activating solution.

[0092] A self-heating treatment bonnet can be similarly devised using versions of the multi-layered material described above. Also, in one of various self-heating bonnet embodiments, a multi-layered disposable material can be utilized, (i.e., a "fabricated sheet"), housing an outer layer and an inner layer, with an intermediate layer of exothermic reactant. Activation of the exothermic reactant is achieved by applying or soaking the exemplary embodiment(s) in an activating solution, such as an electrolyte solution or water, which triggers the exothermic reaction to generate the desired heat. Application of the formulations described above can be made to the interior of the bonnet, or to the skin, prior to use of the self-heating bonnet, if so desired. As mentioned above, other methods, combinations of topical/formulation applications (prior to, after, partially, etc.) can be devised and are understood to be within the spirit of this disclosure.

[0093] The multi-layered disposable material can be such that it can be manufactured from plastic, water proof paper, woven fabrics, foil, natural fibers, and so forth. One layer of the multi-layered disposable material may be of a semi-permeable or even permeable material that permits entry of the activating solution into the exothermic reactant that is embedded or sandwiched therein. For example, an outer layer, inner layer, chamber and (optionally) activation solution entrance aperture can comprise the bonnet. In some embodiments, it may be desirable to have the outer layer as the water proof/limited permeability layer and the inner layer as the permeable material. In some applications, it is contemplated that the user may reverse the "wearing" of the bonnet during mid-treatment to go from a dry heat treatment (impermeable layer being the inner layer) to a wet heat treatment (impermeable layer being the outer layer), or vice versus. Therefore, the user can be afforded either option, if so desired.

[0094] As apparent from the above description, it is possible to configure a bonnet that has both the inner layer and outer layer as impermeable layers (dry heat), whereas venting gases from the exothermic reaction can be channeled out through the solution entrance hole, or other holes to provide venting. Conversely, it is possible to configure a bonnet that has both the inner layer and outer layer as permeable layers (wet heat). A viable modification of such a bonnet would be to have different rates of peπneability for the inner layer and outer layer, thus allowing different levels of wet heat to be applied to the user's head/hair. Another modification is to allow the seals between chambers to be water/solution proof, so as to allow a user to selectively activate different chambers as desired.

[009S] The bonnet may be "pre-configured" with the formulations or "post-configured" with the formulations at the "head-side" layer. In some embodiments, a pre-configured bonnet may require the removal of a seal or exposure of the formulation by some physical action, such as breaking a seal, shaking, and so forth. The seal may be waterproof, if so desired. Also the removal of the seal may occur after the bonnet is exposed to the activating solution.

[0096] A self-heating treatment body strip can be similarly devised using versions of the multi-layered material described above. To activate the self-heating multi-layered "fabricated sheet", the user may simply dip the sheet (with the permeable side exposed) into the activating solution (if using Lava Gel™, then water can be the activating solution) and letting the activating solution soak into the exothermic reactant that is bounded by the inner and outer layers. Some degree of shaking may be needed, to better distribute the activating solution. After a couple of minutes, the self-heating multi- layered fabricated sheet will rise to its desired/designed temperature and maintain itself at that temperature for an extended period of time. Using Lava Gel™, extended heat times of over an hour or more can be achieved.

[0097] In some embodiments, it may be desirable to have both inner layer and outer layer as impermeable layers, using a port or channel in either layer for introducing the exothermic activating solution. In these embodiments, no activating solution need be exposed to the elements (or drip out) after being applied to the self-heating multi-layered fabricated sheet, as well as the fact that with a permeable membrane or layer, heat can escape through the membrane, which could reduce its heating longevity.

[0098] The layers may be formed in strips (laterally alternating), whereby one strip or section of outer layer would be impermeable and another neighboring strip or section of outer layer would be permeable. Thus, different regions of the self-heating multi-layered fabricated sheet would be heated. An example would be the use of a wrap wherein only one section of the wrap would be self-heating, for localized heating (e.g., knee, calf, shoulder, etc.). In other embodiments, it is envisioned that the permeable layer may have an impermeable tear-off covering, and by tearing off appropriate impermeable tear- off cover(s), selected portions or sections of the self-heating multi-layered fabricated sheet would be heated,

[0099] A sleeve may be formed from the exemplary multi-layered fabricated sheet. The exemplary sleeve can be configured with shoulder-side opening, wrist-side opening, inner layer, outer layer and optional seams and optional protective area. The inner layer and outer layer may be any combination of permeable/impermeable materials, as discussed above. A leg or calf or other part of the body wrap may be contemplated.

[00100] Various portions or sections of the strip, sleeve, wrap, etc., may contain the topical/formulations described above, applied in any of the manners/combinations outlined.

[00101] In some embodiments, the strip, sleeve, wrap, etc., may require the removal of a seal or exposure of the formulation by some physical action, such as breaking a seal, shaking, and so forth. The seal may be waterproof, if so desired. Also the removal of the seal may occur after the appliance is exposed to the activating solution,

[00102] It will be understood that many additional changes in the details, materials, steps and arrangement of parts, which have been herein described and illustrated to explain the nature of the invention, may be made by those skilled in the art within the principle and scope of the invention as expressed in the appended claims.

[00103] All references cited herein are incorporated by reference as if each was individually identified to be incorporated by reference. Additionally, the attached Exhibits are individually incorporated by reference herein and are understood to constitute part of the disclosure of this invention.

Claims

CLAIMS What is claimed is:

1. A cosmeceutical composition for topical administration, comprising: a yeast cell lysate;

a fatty acid-peptide conjugate;

a plant stem cell extract;

an enzyme or coenzyme with antioxidant activity; and

a microalgae cell extract.

2. The composition of claim 1 which is formulated into a liposomal delivery system.

3. The composition of claim 1, wherein the yeast cell lysate comprises a tissue respiratory factor.

4. The composition of claim 1, wherein the cosmeceutical composition has an enhanced affect at a temperature above room temperature.

5. The composition of claim 1, wherein the fatty acid-peptide conjugate is an admixture of Palmitoyl-Glycyl-Histidyl-Lysine (PaI-GHK) and Palmitoyl-Glyeyl- Glutaminyl-Prolyl- Arginine (PaI-GQPR).

6. The composition of claim 1, wherein the plant stem cell extract comes from apple stem cells.

7. The composition of claim 1, wherein the enzyme is derived from Thermits thermophilic.

8. The composition of claim 1, wherein the microalgae cell extract is an extract of Dunaliella salina.

9. A method for enhancing delivery of the composition of claim 1 to skin, comprising: topically applying the composition to the skin; and applying a controlled heat to the skin during or after applying the composition.

10. The method of claim 9, wherein heat is applied with an exothermically heated material.

1 1. The method of claim 10, wherein the exothermically heated material is incoiporated into a mitt, bootie, bonnet, mask or body strip.

12. An apparatus for heat-enhanced topical delivery of the composition of claim 1 , comprising: an amount of the composition of claim 1 applied to a skin-side of a self-heating fabricated material,

wherein the fabricated material further comprises an exothermic material, and wherein the exothermic material is activatable by an aqueous solution.

13. The apparatus according to claim 12, wherein the aqueous solution is saline.