BR112014017913B1 - COSMETIC IN THE FORM OF AN EMULSION, KIT AND PROCESS FOR IMPROVING SKIN APPEARANCE - Google Patents

Description

COSMETICS IN THE FORM OF AN EMULSION, KIT AND PROCESS FOR IMPROVING SKIN APPEARANCE

REFERENCE TO RELATED APPLICATIONS [0001] This application is a non-provisional application that claims the priority benefit of provisional application US 61 / 589,592, filed on January 23, 2012, the contents of which are incorporated herein by reference.

FIELD OF THE INVENTION [0002] The present invention in general relates to cosmetics and their components and, in particular, to a cosmetic that contains protein extracts of hatching eggs and their formulations that are useful to improve the quality of the skin.

BACKGROUND OF THE INVENTION [0003] Traditional cosmetic formulations have included various oils and lipophilic substances that formulate a protective layer on the skin to mitigate the effects of bad weather. The inclusion of a sunscreen in a cosmetic also mitigates ultraviolet damage to the skin associated with aging and the production of free radicals. Other conventional components for cosmetic compositions are emollients that transmit appreciable amounts of water to the skin, which in combination with oils affect the amount of moisture released or retained from the skin to disguise wrinkles. Unfortunately, such conventional cosmetic formulations have failed to achieve long-term, chemical improvement in skin quality.

[0004] Observation of fish incubator workers, whose hands had exceptional skin quality, even with prolonged exposure to cold water, was the genesis of the isolation of a new class of proteins associated with fish eggs. Such proteins are detailed, for example, in US 6,346,245; 6,592,866; US 2011/0280882; WO2011 / 06434 and US 2009/0274770. Proteins in isolation included

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2/34 zonases, leucolectins, very acid proteins (VAPS) and chorelisins. At least some of these proteins are effective in treating autoimmune and inflammatory disorders, as well as damaged skin, all with minimal side effects. Although these proteins have improved the function associated with weather-damaged skin and various environmental stressors, such as exposure to air conditioning or air pollutants, these proteins appear to have little or no effect on the other attributes associated with damaged and aged skin. In addition, while the inclusion of such isolated fish roe proteins has been contemplated, the ability to maintain protein activity and provide storage stability in a cosmetic formulation represents a technical challenge.

[0005] Thus, there is a need for a stable cosmetic in storage, which treats the multiple aspects of skin aging, while incorporating the anti-inflammatory attributes of eggs and, in particular, proteins derived from fish eggs. There is also a need for a cosmetic that is essentially free of synthetic cosmetic components that have unwanted deleterious effects on the skin and bioaccumulation problems.

SUMMARY OF THE INVENTION [0006] A composition is provided, which includes a fish roe protein isolate with an anti-inflammatory effect on the mammalian skin. A natural product extract is also present, which includes unsaturated fatty acids and sterols. The extract of natural products has antioxidant activity. An emulsifier or solubilizing agent, or a combination of these methods is provided to form a mixture of the isolate and the extract. A composition is also provided that includes a hatching egg protein isolate with an anti-inflammatory effect on the mammalian skin. At least one biocide of protective activity of the egg protein isolate hatching during 3 months of

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3/34 storage at 20 ° C is present while the activity, when applied to a subject's skin after storage, remains at 80% fresh and unformulated protein. An emulsifier forms a mixture of hatching egg protein isolate that has a buffered aqueous phase at a pH between 5.6 and 7.9. A kit is even supplied with a composition mentioned in a non-sterile, multipurpose packaging, together with instructions for its application on the skin to promote skin rejuvenation.

[0007] A process of producing a cosmetic, including a hatching egg protein isolate having activity on the skin of a mammal including the formation of at least one aqueous phase and at least one hydrophobic phase. An emulsifier and at least one biocide are present in at least one of the aqueous phase and at least one hydrophobic phase. In at least one aqueous phase and in at least one hydrophobic phase, they are combined to form an emulsion. The emulsion, or aqueous phase, is at least buffered at a pH between 5.5 and 7.9 prior to the addition of the hatching egg protein isolate to the emulsion.

[0008] A process to improve the appearance of the skin is provided that includes applying the composition to the skin of a mammal at a level between 1 x 10 ^-6 to 0.1 mg of the isolate and 2 x 10 ^-7 to 0.02 mg of one of the unsaturated fatty acids per square centimeter of skin at least three times a week to achieve improved skin appearance.

DESCRIPTION OF THE INVENTION [0009] The present invention is useful as a cosmetic with skin restorative attributes. In one embodiment, a cosmetic of the invention is formulated to provide a synergistic effect between the various components. In yet another embodiment, a cosmetic composition of the invention is formulated without synthetic components. A cosmetic, in all embodiments of the invention, maintains the activity of protein components, having storage stability at room temperature, and

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4/34 in response to the microbial challenge through non-sterile processing and use subject to microbial contamination. A formulation process is provided that provides a cosmetic formulation with these attributes of maintained protein activity and storage stability under the conditions of use and storage of cosmetics.

[0010] In accordance with the present invention, active hatching egg proteins and, in particular, isolated fish roe protein with anti-inflammatory effect on mammalian skin are combined with a natural product extract containing unsaturated fatty acids and at least one sterol. The extract has, simultaneously, an antioxidant activity and a collagenase inhibiting activity and is combined with an emulsifying agent that stabilizes the mixture containing the isolated protein and the extract. It is appreciated that several extracts are optionally used here simultaneously to achieve the requirements of composition and activity. It has surprisingly been discovered that the activity of hatching egg proteins, and in particular fish roe protein and natural products, work synergistically in some embodiments of the invention to achieve the superior appearance of the skin and, in some embodiments, skin rejuvenation. , compared to any ingredient used alone in a controlled formulation. In still other embodiments, the storage stability of a cosmetic of the invention for at least 3 months at 20 ° C is provided while the protein component activity, when applied to the subject's skin after storage remains within 80% of the non-protein formulated and recently isolated.

[0011] A fish roe protein isolate, for example, includes a zonase, a leucolectin, or a combination of these and has the properties described in US 6,346,245, col. 4, line 15 col. 5, line 8; US 2009/0274770 A1 [0321] - [0326] and US 2011/0280882 [0157] - [0194]; crude extracts that contain these proteins, and combinations of these.

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Exemplary sources of fish eggs spawn for protein isolation include sturgeon roe, salmon, white fish, coregono, cod, whiting, and burbot. It is appreciated that other sources of egg proteins operative here include amphibian egg coatings; such as tadpoles, and salamanders; cases of reptile eggs, cases of bird eggs. Typically, an egg - or in particular, the isolated fish roe protein containing at least some of the above proteins constitutes 0.0001 to 10% of the total weight of a cosmetic formulation of the invention with 0.01 to 10 percent of the extract of protein to be active. It is preferred that the amount of protein present in a cosmetic of the invention depends on factors, including illustratively, miscibility with other components of another cosmetic, and degree of damage to the skin.

[0012] It is to be understood that in cases where a range of values is provided since the set is intended to cover not only the end point values of the range, but also the intermediate values of the range as being expressly included in the range and varying by the last significant figure of the interval. For example, the range quoted from 1 to 4 is intended to include 1-2, 1-3, 2-4, 3-4, and

1-4.

[0013] A composition of the invention also contains a natural product extract. The natural product extract includes an unsaturated fatty acid and a sterol, in an amount to provide an antioxidant activity and collagenase inhibition activity greater than 10% compared to the control, as measured by the Oxygen Radical Absorption Capacity (ORACFL) through tests and lipoperoxidation marker in culture medium with an extract of natural product present in 1% and irradiated with ultraviolet B radiation in an amount of 150 mJ per square centimeter, respectively. R.L. Prior, et al. Assays for Hydrophillic and Lipophillic Antioxidant Capacity (oxygen radical absorbance capacity (ORACFL)) of Plasma and other Biological and Food

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Samples. J. Agric. Food Chem. (2003) 51: 3273-3279. It is appreciated that natural extracts often contain mixtures of saturated fatty acids and one or more sterols, and other components of different dermatological activities. An extract product used in a composition of the invention also inhibits collagenase activity in some modalities, more than 5% and some modalities, as measured against a control for an extract of 0.1%, by weight, in the control using a protocol visual measurement of the anti-collagenase test in human biopsies with Cyro skin slices to be incubated 30 minutes at 37 ° C, with and without an extract followed by three hours of incubation at 37 ° C without (negative control) or with 15 microliters per milliliter of collagenase followed by washing of biopsy samples followed by staining and microscopy for visual measurement of Masson's trichomes. In some embodiments, a natural product extract used in a composition of the invention includes at least omega-3, omega-6 and omega-9 fatty acids. In still other embodiments, unsaturated fatty acids are present in the extract of natural product in an amount of at least 20 milligrams per 100 grams of extract. A natural product extract of the invention also includes at least one type of sterol, sterol illustratively including cholesterol, campesterol, stigmasterol, sitosterol, hydroxylated forms and combinations thereof. In some embodiments, at least one sterol is present in a natural product extract in an amount greater than 50 mg per 100 g of extract. Typically, a natural product extract is present in the composition of the invention from 0.01 to 10 percent by weight and, in some embodiments, from 0.1 to 3 percent by weight. Determining factors in the choice of the amount of natural product extract in an illustrative way include stabilization of egg proteins in hatching cosmetics and glycosides in the presence of egg proteins in hatching, degree of skin damage and useful life of cosmetics.

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7/34 [0014] A natural product extract according to the present invention is easily formed by conventional techniques such as the extraction of a part of a plant, such as leaves, stems, buds, roots, fruits, and combinations thereof with a first solvent that is either hydrophilic or hydrophobic and filtering the mixture after being in contact with the solvent for times of 1-48 hours, the amount of solvent is usually between 1 and 20 volumes relative to that of the plant material. Extraction by hydrophilicity-reducing solvents and elution by conventional techniques allows one to produce and extract an inventive natural product. Representative fatty acid / sterol extraction techniques are found in J.S. O'Brien and A.A. Benson, J. Lipid Res., 5, 1964, pp 432-436. The organic solvent illustratively includes ethers, furans, petroleum distillates, alkanes, halogenated alkanes, alcohols, acetates and combinations thereof. After sufficient interaction between the phases, typically between 20 minutes and 48 hours, the organic phase is isolated to obtain an extract according to the present invention. Suitable sources of extracts include red, green, brown algae and tuft-forming cyanobacteria collectively referred to as “algae” and plants or components that have the same, purple fruits, black red, orange or leaf colorations where the leaf coloring is measured in the presence or absence of chlorophyll. Seaweed represents a common source of natural product extracts of the invention with high levels of unsaturated fatty acids and sterols. It is appreciated that synthetic and / or animal sources of unsaturated fatty acids and sterols are readily used in a composition of the invention.

[0015] In addition to the hatching egg protein isolate and the natural product extract, the inventive composition in some modalities includes one or more other components normally used in cosmetic products, with the proviso that these components maintain the isolate attributes hatching egg protein and that

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8/34 the extracts of natural products are maintained. Representatives of conventional cosmetic components, which are easily formulated here in an illustrative manner include optionally deionized water constituting the bulk by weight of the inventive cosmetic; petroleum solids and waxes, such as lanolin, ceresin, microcrystalline wax, carnauba wax, C10-C30 fatty acids, C10-C30 alcohols, oils such as olive oil, jojoba oil, castor oil, sunflower oil, sunflower oil hazelnut, shea butter, safflower oil, and viscosity modifiers / stabilizers such as guar gum, xanthan gum, carrageenan, hydroxyethyl cellulose, sclerotia gum, carbomers, polyacrylates, and Cw-C30 acrylate acrylate copolymers; hydrophilic moisturizers, such as glycerin, propanediol, sodium lactate, sodium PCA, emollients such as dicaprilil ether, myristyl myristate, C10-C30, C10-C30 caprates, containing caprylic triglycerides and / or capric acids, and C10- C30 glycerides; coemulsifiers, such as cetyl alcohol, stearyl alcohol, glyceryl stearate, stearic acid, glycol stearate, glyceryl distearate, and emulsifiers such as nonionic surfactants and anionic surfactants, including illustratively lactylate, caproyl lactylate salts, cocoyl-lactylate, stearoyl-lactylate, cetearyl olivate, sorbitan olivate, glyceryl oleate, in which the typical cation citrate includes sodium, potassium, ammonium, calcium and magnesium salts, silicone oils such as inorganic dyes and accompaniments inorganic pigments, metal-treated or silicon-treated inorganic or organic pigments and organic dyes, non-ionic surfactants, polysiloxanes, preservatives, water pH modifiers, vitamins and vitamin metabolites such as vitamin E, vitamin K, or complexes vitamin B, essential oils; ultraviolet light absorbing compounds, perfumes, moisturizers, improving blood flow agents, inducing agents a feeling of cold; antiperspirants; active, and biocides.

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9/34 [0016] As used here, an asset is defined as an organic compound applied to human skin to cleanse, beautify, promote attractiveness, or change the appearance that is effective in improving skin attributes, regardless of the fact that the compound is polar or non-polar in nature. As used herein, this term specifically excludes a hatching egg protein, a mixture of hatching egg proteins, and a natural product extract. Active organic compounds operative here illustratively include additional extracts, such as Songyi mpo mushroom, Acacia farnesiana, Euterpe oleracea, Malpighia glabra, Bixa Orellana, Prunus dulcis, Aloe vera, rhododendron ferrugineum, Amaranthus caudatus, Angelica archangelica, Pimpinella domisumum , Mentha suaveolens, avenanthramide oats, Prunus armeniaca, Arnika montana, Cynara scolymus, Asparagus officinalis, Persea americana, Cardiospermum halicacabum, Melissa officinalis, Bambusa vulgaris, Musa paradisiaca, Adansonia digitata, Berberis vulgaris, Epimus, Basilimum ursinum, Geum urbanum, Betula pubescens, Quassia amara, Nigella sativa, Ribes nigrum, Morus nigra, turnip, Camellia sinensis, Rubus fruticosus, Iris versicolor, Vaccinium myrtillus, Borago officinalis, Vicia faba, Menyanthes trifentum, Escagulum, Fagia, Ruscus aculeatus, Theobroma cacao, Acorus calamus, Carum carvi, Elattar ia cardamomum, Daucus carota sativus, Erythroxylum catuaba, Centaurium erythraea, Matricaria chamomilla, Prunus cerasus, Cicer arietinum, Chlorella vulgaris, Aronia melanocarpa, Cinchona pubescens, Cinnamomum verum, Syzygium aromaticum, Cocosea, Barbados, Cocos officinale, Plantago major, Coriandrum sativum, Zea mays, Papaver rhoeas, Centaurea cyanus, Gossypium herbaceum, Vaccinium vitis-idaea, Primula veris, Vaccinium macrocarpon, Cucumis sativus, Curcuma longa, Bellis perennis, Rosa damascena, Turnera diffus, Taraxic diffuse, Tara diffuse, Taraxum diffuse

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10/34 dactylifera, album Lamium, Rosa canina, Hylocereus undatus, Daemonorops drake, Echinacea angustifolia, Leontopodium alpinum, Sambucus nigra, Acanthopanax senticosus, Phyllanthus emblica, Eucalyptus globulus, Oenthera biennium, Euichrysumumen, Helichrysumenen, Helichrysumen carica, Linum usitatissimum, Plumeria alba, Fumaria officinalis, Rumex acetosa, Gardenia Yessminoides, Gentiana lutea, Zingiber officinale, Ginkgo biloba, Panax ginseng, Solidago virgaurea, Vitis vinifera, Citrus paradisi, Camellia sinensis, Paullinia cupana, grullis flana Lawsonia inermis, Hibiscus sabdariffa, Lonicera japonica, Humulus lupulus, Aesculus hippocastanum, Equisetum arvense, Sempervivum Tectorum, Hyssopus officinalis, Cetraria islandica, Baptisia tinctoria, Chondrus crispus, Communismis, Hedera helix, Jasmine , Actinidia chinensis, Pueraria lobata, Alchemilla vulgaris, Lavand ula angustifolia, Citrus limon, Cymbopogon cytratus, Glycyrrhiza glabra, Lilium candidum, Citrus aurantifolia, Tilia cordata, Nelumbo nucifera, Macadamia ternifolia, Magnolia biondii, Berberis aquifolium, Malva sylvestris, Ulifea, Mangifera indica, Calendula, Mangifera indica, Calendula , melon, Silybum marianum, Panicum miliaceum, Colophospermum mopane, Sorbus aucuparia, Verbascum thapsus, Guazuma ulmifolia, Myrtus communis, Tropaeolum majus, Azadirachta indica, Urtica dioica, Ascophyllum nodosum, Myristica fragrans, Quercustica fragrans, , Olea europaea, Allium cepa, Citrus sinensis, Mentha citrata, Origanum vulgare, Viola tricolor, Carica papaya, Capsicum annuum, Petroselinum crispum, Passiflora incarnata, Prunus persica, Pyrus communis, Centella asiatica, Paeonia officinalis, Piper nigrum, Miperha comosus, Prunus domestica, Punica granatum, Pulmonaria officinalis, Cucurbita pepo, Cydonia oblong a, Rubus idaeus, Trifolium pratense, Oryza sativa, Aspalathus linearis, Rosa centifolia, Rosmarinus officinalis, Secale cereale,

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Salvia officinalis, Schisandra chinensis, Hippophae rhamnoides, Sesamum indicum, Capsella bursa pastoris, Albizia julibrissin, Artemisia abrotanum, Mentha spicata, Triticum aestivum, Spirulina platensis, Picea abies, Hypericum perforatum, anthelia, Helminthia, Fragia, Stevia, Stevia , Tanacetum cinerariifolium, Citrus reticulata, Thymus vulgaris, Salvadora persica, Valeriana officinalis, Vanilla planifolia, Verbena officinalis, Viola odorata, Juglans regia, Nasturtium officinale, Citrullus lanatus, Triticum aestivum, Thymia salisis, Thymia salensis, Crimea, Salisbury virginia, Isatis tinctoria, Lycium barbarum, Artemisia absinthium, Achillea millefolium, Hymenaea courbaril, Cananga odorata, Tricholoma matsutake Singer, or their combinations. It is preferred that such extracts may also have a certain degree of antimicrobial activity which, in certain embodiments, increases the effectiveness of the biocide. Active organic compounds are typically used in a formulation of the invention from 0 to 1 percent by weight and, in some embodiments, up to 5 percent by weight. Although these active agents tend to be more tolerant of conventional cosmetic processing conditions and in many cases they are liable to be included in an aqueous or lipophilic phase other than hatching egg protein and natural product extract, which should be appreciated than organic compounds such assets in one embodiment are added after the formation of the emulsion through phase C or another, separately to the emulsion.

[0017] An inventive cosmetic is easily formulated into a water-based cosmetic in an illustrative way including a toner, a spray, a serum, a cream, a mask, a sunscreen, a cleanser, a moisturizer, a bath preparation, a liquid foundation, an astringent, a shaving preparation, a hair care product, and other skin care products, or a topical medicine illustratively including an anti-inflammatory agent. In a particular embodiment of the present invention, a cosmetic is provided that includes a

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12/34 plant-based emulsifier, such as a lactylate that is essentially free of silicone, oil, petrolatum, and free of synthetic parabens. An "substantially free" emulsifier of these compounds is defined herein as having less than 10 weight percent emulsifier of these compounds in total. An inventive formulation is easily applied to human skin in regions that, specifically, and illustratively include the face as a whole; the regions of the face around the mouth, eyes, forehead, temples, cheeks, chin, hand or foot; neck, scalp; elbow; chest; forearm, shoulders, legs, and their combinations.

[0018] An inventive cosmetic presents difficulties in the formulation to maintain the activity of the protein. In general, it is observed that heating and high pure mixture; Common cosmetic formulation techniques tend to denature the active protein. In addition, active proteins are prone to microbial degradation, especially when a cosmetic of the invention is processed in non-sterile conditions and stored at an ambient temperature of 20 ° C. Formulation of an inventive cosmetic requires that the hatching egg protein be exposed to a limited range of pH values, pure mixtures and heat during the formulation process, in order to keep the hatching egg protein present in it. To maintain hatching egg protein activity, a cosmetic formulation process includes buffering any cosmetic constituent phase to a pH between 5.6 and 8, before introducing proteins to incubate the eggs at that stage. Buffering is commonly practiced in the field and includes the addition of an acid, a base, an acid salt, or a combination of these to balance an aqueous solution or aqueous phase of an emulsion at a desired pH. It is appreciated that the pH measurement is routine and measured quantitatively by means of volumetric titration or with potentiometric electrodes, such as

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13/34 commercially available from Thermo Fisher Scientific (Waltham MA, USA), and measured quantitatively with litmus paper kits.

[0019] While the pH values of an inventive cosmetic are adjusted between 5.6 and 7.9 for the maintenance of protein activity partially inhibits bacterial degradation of hatching egg proteins, biocides are present in a formulation of the invention to maintain the activity of the protein in response to organisms present during formulation and in response to the continuous sowing of the cosmetic with several microbes during the use of a multiple dose container through topical and respiratory contact, with an inventive aesthetic. As a result, a cosmetic formulation of the invention, in stable formulations for multiple use and storage, includes a broad spectrum biocide or a combination of biocides. A broad-spectrum biocide is defined herein as having activity against Gram-positive bacteria, Gram-negative bacteria and fungi associated with a healthy human subject. Biocides that operate here illustratively include benzoic acid, parabens, salicylic acid, carbonic acid, sorbic acid, alkyl p-hydroxybenzoates, p-chlorometacresol, hexachlorophene, benzalkonium chloride, chlorhexidine chloride, trichlorocarbanilide, phenoxyethanol, acylar , glutathione, malic acid, tartaric acid, ascorbic acid, ascorbates, essential oils from plants, mutacin proteins, and their combinations. In certain embodiments of the present invention, only the derived biocides are naturally present. Naturally derived biocides illustratively include fermentation filtrates, such as Lactobacillus, Streptococcus mutans, and Leuconostoc; bisabolol, eucalyptol, thymol, inositol, saponins, and natural plant extracts, such as Japanese honeysuckle (Lonicera japonica), Yangti (Rumex japonicus), Kushen (Sophora flavescens), candock, wild oregano, orange, sage, manifold, mallow , chuanxiong (Cnidii officinale Makino), Japanese green gentian (Swertia japonica Makino), bisabolol, thyme, danguii (Angelica sinensis),

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14/34 orange peel, birch, horsetail, dishcloth gourd, horse chestnut, saxifraga (Saxifrage stolonifera), arnica, lily, mugwort, peony, aloe, gardenia, as well as those detailed in MM Cowan, Clinical Microbiology Reviews, 12 ( 4) Oct. 1999, p. 564-582, or combinations thereof. These extracts are obtained by procedures detailed in Clinical Microbiology Reviews, 12 (4) Oct. 1999, pages 573-574 and, often using a hydrophilic organic solvent such as an alcohol C1-C8, polyhydric alcohols, water and aqueous alcohols . In a specific embodiment of the Japanese honeysuckle, floretine (apples), galangin (Helichrysum aureonitens) are used as naturally derived biocides, each alone or in combination. It is appreciated that, in addition to the biocidal properties, a natural extract often gives a fragrance to an inventive cosmetic. A biocide is present in multipurpose cosmetic packaging, in specific embodiments of the invention from 0.1 to 5 percent in total weight. It is appreciated that amounts of biocides in excess of 5 percent by total weight are readily included as desired.

[0020] A cosmetic of the invention is in certain modalities of pH buffered between 5.6 and 7.9 and in still other modalities between 6.5 and 6.8, while in other modalities still between 6.8 and 7.3. Without wishing to be bound by a particular theory, these pH values facilitate the stabilization and storage stability of egg extract such as isolated fish roe, such as and natural product extract components. The formulation of a cosmetic of the invention is facilitated by the pre-formulation of at least two components that include an oil-in-water emulsion, and a second phase of a water-in-oil or silicone emulsion with optional additional phases, such as , for example, a third phase that contains the active egg extract, such as isolated fish roe and / or natural product extract. Alternatively, it is appreciated that one or both of the hatching egg isolate or extract of natural products are easily dispersed either in the oil emulsion in

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15/34 water, water in oil, water in silicone emulsion or a serum formulation. It was surprisingly found that the pH buffering of several phases, before the addition of the active egg protein extract is crucial for establishing the storage stability for the cosmetic of the resulting invention, as well as the activity of the skin protein in question.

[0021] The components of a minority phase of an oil-in-water emulsion illustratively include oils, waxes, esters, ethers, butters, glycerol, or combinations as detailed above. The components of a major phase oil-in-water emulsion include water, aqueous salts, humectants, and various water-soluble materials, as specified above. An emulsifier of this type of system includes ionic surfactants, non-ionic surfactants, as specified above, and also including lecithin, liposomes and oleosomes. Other non-essential additives are readily provided as detailed above, which include viscosity modifiers, emulsion stabilizers, flavors, and the like. In specific embodiments, the oil-in-water emulsion has a pH value of 5.6-7.9. In specific embodiments, the oil-in-water emulsion has a pH value of 6.5 and 6.8.

[0022] The water-in-oil or water-in-silicone emulsion varies in the relative amounts of the components of the minority and majority phase and is preferably based on non-ionic surfactants and silicone derivatives as emulsifiers. Viscosity modifiers and / or emulsion stabilizers, illustratively, include water-soluble salts, resins, waxes, and their combinations, as detailed above. In specific embodiments, the internal phase is the pH adjusted to a pH value of 5.6 to 7.9. In certain embodiments, the internal phase pH value is 6.5 to 6.8. It is preferred that various vitamins such as vitamin E, vitamin K, ascorbic acid derivatives such as ascorbic acid glucosides are

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16/34 readily supplied in one or both of the aforementioned emulsion phases or supplied as a separate third phase.

[0023] A cosmetic of the invention is easily formed by a combination of premixed phases, as described above or, alternatively, all these components are mixed in a conventional sequence in the art as a unified mixture. A process for improving the appearance of the skin is envisaged that includes applying a composition of the invention to the skin of a mammal at a level of 1 x 10 ^-6 to 0.1 mg of isolate and 2 x 10 ^-7 to 0.02 mg of unsaturated fatty acids per square centimeter of skin at least three times a week to achieve improved skin appearance.

[0024] A summary of the typical ranges for the cosmetics of the invention is provided in Tables 1-3. It should be appreciated that other conventional base formulations are easily adjusted, as detailed here to stabilize hatching egg proteins, either alone or in combination with a natural product extract containing fatty acids and sterols. A variety of base formulations here conventional for a cosmetic are provided in EW Flick, “Cosmetic and Toiletry Formulations”, 2 ^nd Ed., Vol. 8; 2001, Noyes Publications, Norwich, New York, USA (ISBN 0-1855-1454-9).

[0025] Table 1. Typical intervals of the inventive cosmetic in oil-in-water emulsion, where the percentages are total percentage by weight. Three phases and four phase versions are provided here.

[0026] Three-phase formulation

Phase A-1

Deionized water Remaining Viscosity modifier / stabilizer 0 to 5% Hydrophilic moisturizer 0 to 20% Vegetable oil / butter 1 to 20% Vegetable-based emollient 0 to 20%

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Coemulsifier 0 to 10% Secondary emulsifier 0 to 10% Lactylate emulsifier 0.1 to 10%

PH adjustment / buffering to pH 5.6 to 7.9 *

Biocide 0 to 5% Fragrance Non-polar active organic compounds 0.1 to 10% 0 to 5% 0 to 1%

Phase B-1

Deionized water Active (L-Ascorbic acid 2-Glucoside) Citric acid 1 to 20% 0 to 10% 0 to 1% Sodium citrate dihydrate 0 to 1% Sodium Hydroxide (20%) to pH 5.6 to 7.9 Polar active organic compounds Phase C-1 0 to 5% 0 to 1% Active (0.034% egg hatching protein) Active (natural product extract) Fourth stage formulation Phase A-2a Deionized water Viscosity modifier / stabilizer 0.001 to 10 0.001 to 10 Remaining 0 to 5% Hydrophilic moisturizer 0 to 20%

PH adjustment / buffering to pH 5.6 to 7.9 * Biocide ** 0 to 5% Fragrance Non-polar active organic compounds 0 to 10% 0 to 5% 0 to 1% Phase A-2b Vegetable oil / butter 1 to 20%

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Vegetable-based emollient 0 to 20% Coemulsifier 0 to 10% Secondary emulsifier 0 to 10% Lactylate emulsifier 0.1 to 10% Biocide ** 0 to 5% Fragrance 0 to 5% Non-polar active organic compounds 0 to 1% Phase B-1 Deionized water 1 to 20% Active (L-Ascorbic acid 2-Glucoside) 0 to 10% Citric acid 0 to 1% Sodium citrate dihydrate 0 to 1% Sodium Hydroxide (20%) to pH 5.6 to 7.9 0 to 5% Polar active organic compounds 0 to 1% Phase C-1 Active (0.034% active protein) 0.001 to 10% Active (natural product extract) 0.001 to 10%

* - pH adjustment can also be performed after the combination of Phases A and B, however, before the addition of Phases C-1.

** - Some amount of biocide is required, typically between 0.1 and 5 percent in total weight in non-sterile multipurpose formulations.

Table 2. Typical values of the cosmetic invention in silicone water emulsion, where the percentages are total percentages by weight.

Phase A

20% polysiloxane

Other silicone 1 to 10% Emulsifying / surfactant silicone 1 to 15% Emollient (non-silicone) 1 to 10% non-polar active organic compounds 0 to 1% Phase B

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19/34

Deionized water Glycol remaining 1 to 30% Alkali metal salt 0.2 to 2% Viscosity modifier / stabilizer 0.1 to 3% PH regulator Biocide ** 0 to 3% 0 to 2% Active polar organic compound Phase C 0 to 1% active (0.034% active protein) active (natural product extract) 0.001 to 10% 0.001 to 10%

Table 3. Typical ranges of inventive cosmetics in serum formulation, where percentages are total weight by weight

Phase A Deionized water Viscosity modifier Remaining 0.1 to 2% PH regulator Glycol 0.1 to 3% 0.01 to 10% Glycerin 0.01 to 10% Biocide 0.01 to 2% Polar active organic compound Phase B 0 to 1% Solubilizer 0.1 to 10% Non-polar active organic compound Phase C 0 to 1% active (0.034% active protein) active (natural product extract) 0.001 to 10% 0.001 to 10%

[0027] The present invention is further illustrated with reference to the following non-limiting examples.

EXAMPLE 1

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20/34 [0028] A cosmetic cream is prepared after the formulation of the three phases. Phase A is heated to 75-80 ° C, with sufficient agitation to form a homogeneous mixture. Phase B is pre-mixed in a separate vessel, at room temperature and added to Phase A, at 40 ° C or lower. Phase C is added to the one now combined with mixing phases A and B in a manner with sufficient mixing until a homogeneous cream is formed. Thus resulting in a homogeneous and stable storage cream.

EXAMPLE 2 [0029] A cosmetic cream in which the protein isolated from fish eggs and a natural product extract are components of Phase A which is provided, instead of the isolated fish egg protein the natural product extract present in a phase separated C, for example, 1. With phase A to be formed at room temperature to avoid denaturing the protein, thus obtaining a cream.

COMPARATIVE EXAMPLES [0030] A comparative composition with Example 1 was created as an isolated fish roe-free cream (Comparative of Example A). Another composition is devoid of a natural product extract (Comparative example B). The controlled formulation is free of both isolated fish roe protein and natural product extract (comparative Example C). A study group of 40 subjects is divided into four groups, each group with daily application of a cream from Example 1, or one of the comparative Examples AC for 12 weeks. The basis of skin elasticity is measured for each subject and at the conclusion of the study after normalizing the control of Comparative Example C, the composition of Example 1 shows superior Elasticity and smoothness compared to Comparative Examples AC. All compositions are normalized to a level between 1 x 10 ^-6 mg of the isolate and (if present)

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21/34 of 2 x 10 ^-7 mg of unsaturated fatty acids (when present) per square centimeter of skin per application.

EXAMPLE 3 [0031] Salmon eggs (Salmo salar) harvested fresh, unfertilized, from females in the reproductive phase (autumn), are kept on ice, and the extract is preferably made immediately. It is possible to freeze dried eggs in a cryoprotectant (for example, 1.5 M of 1,2-propanediol and 0.2 M of sucrose) without breaking the egg membrane. Freezing should be gradual (1 ° C / min) to -80 ° C. The eggs must be defrosted and kept on ice throughout the extract preparation process.

[0032] The eggs are washed twice in HBSS or with sea water, with protease inhibitors (10 pg / ml). The washing solution is removed and the eggs are lysed and homogenized in a pre-cooled glass-to-glass homogenizer Dounce. The lysate is transferred to Beckman Ultra Clear polymer centrifuge tubes (5 ml), while avoiding the transfer of egg coatings, and centrifuged for 15 min at 15,000 g at 4 ° C, in a Beckman ultracentrifuge using a SW55T1 rotor. Three fractions are thus obtained; upper lipid fraction, cytoplasmic medium fraction, and a lower fraction containing egg coatings and nucleic remains. The cytoplasmic fraction of the medium is the collected extract. This extract must contain more cytosolic organelles including mitochondria, lysosomes and peroxisomes, must be clear and viscous, and orange in color. Protease inhibitors (10 µg / ml of storage) are added and the extracts are kept at -80 ° C. Embryos of zebrafish in medium blastula are collected, liquid removed and frozen at -20 ° C to prepare the extract, the embryos are thawed on ice, lysed and homogenized by Dounce glass-glass homogenizer, in a small amount of sea water or HBSS (preferably less than 50% liquid v / v). The lysate is filtered through a sterile linen cloth and centrifuged at 5,000 g

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22/34 at 4 ° C for 20 minutes on an SX4250 rotor using a Beckman X-22R centrifuge. The cytoplasmic extract (supernatant) is collected and protease inhibitors (10 µg / ml) are added. The extract can be filtered by Millipore (0.22 micron MilliQ sterile filter). The extracts are kept at -80 ° C, the pH is measured by litmus paper extract, the protein concentration measured by Bradford assay, and osmolarity measured by an osmometer.

[0033] This general procedure is useful for preparing extracts of sea urchin, shrimp, fish eggs / roe or frog eggs. Briefly, roe collected from pregnant female fish shortly after they released their eggs in a spawning program (injected hCG hormone (1 ml / kg) 6 to 8 hours before the egg is released, usually at dawn (2-4 in the morning), or from pregnant frogs. Roe / eggs are freeze-dried or frozen at -20 ° C or used fresh. The roe is collected from different types of fish. For sea urchin, 0.5 M KCl is injected around the mouth to evoke egg spillage.The extract is prepared from eggs / roe by crushing (cell cracker or dounce homogenization) or centrifugation at different speeds to separate the cytoplasm with all the contents, with / without shell egg (pellucid zone), with / without nucleus / cytoplasm, with / without organelles, with / without lipids.Additional fractionation can be carried out to isolate one or more mRNA proteins, small peptides, carbohydrates and lipids. fatty acids s in the roe are oleic acid, linoleic acid, and omega-3 fatty acids.

[0034] After applying the aforementioned protocol, for salmon egg extracts, salmon egg extracts had, surprisingly, a high protein concentration ranging between 100-380 mg / ml, at a pH between 6.4- 6.8, and an osmolarity of about 350 mOsm. The extracts are clear and viscous and non-filterable (by filter

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23/34

0.45 micron MilliQ). The protein in the extract is easily precipitated by adding water or hydrated solutions with low buffering capacity, due to the high protein content and low pH. Extracts can be neutralized to a pH of 7.0 by adding alkaline (extract of 1-3 μΙ 1M NaOH / ml), after which dilution in water and hydrated solutions is possible. Zebrafish extracts had a protein concentration ranging between 23-26 mg / ml, at a pH between 6.4-6.8, and an osmolarity between 80-150 mOsm. The extracts are clear and non-viscous, filterable and easily diluted with water at all dilutions.

EXAMPLE 4 [0035] Salmon zones purified by gel filtration, plus affinity purification can be further purified to the degree of purity sequence by a final chromatography procedure. This process employs a PBE94 column, with a Tris-acetate buffer (10 mM, pH 9.0), in which subsequent elution is with a salt gradient (up to 1 M NaCl of salt) in this buffer. This step alone increases the catalytic activity of the zones by an additional 7.6 times, for a total purification of 714 times, and with a 28% yield from the starting material. This purification step leaves the zonase protein identity intact as a 28 kDa fraction. Thus, the step does not remove the main unbound protein contaminants from the zonase preparation. The molecular weight of the purified zonases is the same, as observed by the Western blotting technique for zonase portions present in the incubation liquid and in the "crude zonase".

[0036] What seems to happen in the third and last chromatographic procedure is that the small peptides, contaminants are removed. These peptides appear to be oligopeptides with about a dozen residues, probably from the eggshell and / or the salmon embryo. These peptide contaminants appear to have inhibitory effects on zonase catalysis, since their removal

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24/34 increases the catalytic activity of zonase. In addition, its presence interferes with the early stages of Edman sequencing of this zonase product. The two forms of zonases seen in this third purification step are linked in a slightly different way to the column matrix. However, both forms have similar amino acid sequences in their N-terminal portions.

[0037] The partial amino acid sequences of generated CNBr peptides established the zonases as distinct proteins. Structural analysis provided indications that zonases may have distinct catalytic domains and binding substrate, which may be responsible for their sensitivity to calcium chelating agents that act on macromolecular substrates (physiologically) (binding to inhibit, so catalysis is indirectly inhibited) , and also the sensitivity of serine protease inhibitors acting on small substrates (catalysis is directly inhibited).

EXAMPLE 5 [0038] Both zonase and leucolectin are purified from salmon incubation fluid. To improve the protein concentration of the incubation fluid, salmon eggs are transferred to minimal volumes of water before hatching. Highly synchronized hatches can be induced by high temperatures (ambient), or by deoxygenation (Oppen-Berntsen et al. 1990, Aquaculture, 86, pp 417-430), which produces a small amount of the highly concentrated preparation of crude zonase and proteins associated companies.

[0039] The initial purification of zonase involves filtration of salmon eggs incubated through a cloth. This filtrate can be frozen for years without any significant degradation of the zonase, before being thawed and used for further protein purification. This fact considerably facilitates the production of a starting material for

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25/34 the purification of salmon zonase and associated proteins, including leucolectin.

[0040] The next, optional step involves adjusting the filtered protein to 4M urea, to dissociate fragments from the salmon eggshell, which allowed its removal, along with foreign debris by low speed centrifugation (15,000 g; two times for 15 min). This material did not show any sign of clogging columns, which is characteristic of raw materials prepared in a different way than described above. This crude protein preparation is suitable for purification using conventional chromatographic techniques.

[0041] Hatching fluid leucolectin can be isolated in conjunction with zonase. From partially purified zonase preparations (as described above), leucolectin can be isolated by exclusion chromatography such as zonase in its native form which is substantially greater than Leucolectin. For a first separation, Superdex 16/60 columns are used, after which zonase can be removed by affinity chromatography on Benzamidine Sepharose columns.

[0042] For large scale preparations the use of ultra filtration is also suitable as zonase in its native form does not significantly penetrate ultra filters with an exclusion size of 100 kDa unlike leucolectin.

[0043] The buffers used are millimolar Tris (for example, 10 mM), at a pH close to neutrality or slightly alkaline (pH 7.5-8.5), containing 5 mM NaCl. The leucolectin protein is found to co-purify with zonase. The size of this new protein, estimated by chromatography under native conditions, is only timidly 30 kDa.

[0044] The estimated molecular weight of lectin is about 2530 kDa. The estimated pi for the corresponding salmon lectin is about

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26/34 pH = 6.5. The observed pis (Riste, unpublished) are pH 6.5-6.9 in salmon periviteline lectins, and from pH 6.4-6.6 in salmon leukocyte lectins (lectin is identified by means of Western blotting techniques).

EXAMPLE 6 [0045] A study to evaluate the safety and effectiveness of cosmetic products, in the form of a day cream and an eye cream using the formulations described above over a 12 week study period. A total of 40 female individuals, aged 37-60 years are selected for the study. All 40 participants completed the study.

[0046] Under the conditions of this study, the test materials identified as day cream and eye cream demonstrated the potential to improve fine facial lines, wrinkles, hyperpigmentation, uniform skin tone, skin clarity, sagging skin. skin, and tactile roughness over a 12-week period of use. In addition, the test materials are well tolerated and have not demonstrated the potential to cause significant skin irritation when applied to the face.

[0047] Statistically significant improvements in the Packman and Gans scores for the superficial facial lines of the face and the Visual Analogue Scale (VAS) score in facial hyperpigmentation, uniformity of skin tone, skin clarity, sagging skin, and tactile roughness are observed after two, six and twelve times a week of two daily applications. No statistically significant increases in irritation (erythema, edema, dryness, burning, burning and tugging / dryness) from baseline are seen at any post-treatment interval.

EXAMPLE 7 [0048] A study to evaluate the effectiveness of biocide preservatives against microbial contamination in cosmetic products in

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27/34 form of a daily cream and eye cream using the formulations above disclosed during two periods of four weeks of study is carried out. The methods employed are CTFA Microbiological Guidelines, Section 20, M-3, A Method for Testing the Preservation of Miscible Water Hygiene Products and USP 34, Section 61, Neutralization / Removal of Antimicrobial Activity. Test organisms included Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger. For product category 2, the preservative is considered effective in the sample examined in the case of a). The concentrations of viable bacteria demonstrated at least a 3.0 log reduction (99.9%) of the initial count at 7 days, and not for the increase in the duration of the test period and b). The concentration of viable yeasts and molds demonstrated at least a reduction of 1.0 log (90.0%) from the initial count at 7 days, and not to increase the duration of the test period.

[0049] The results of the study showed that day cream and eye cream conformed to the acceptance criteria for product category 2.

EXAMPLE 8 [0050] A study is conducted to determine the comedogenic potential of test materials, in the form of day cream and eye cream, with the formulations described above, determining whether the test materials caused microcomponents after four weeks of repeated application under conditions of occlusive dressing, in relation to a negative control (dressing without dose). Lanolin Acetylated Alcohol served as positive control material in order to validate the study. The tests are carried out over a 4-week study period. A total of 15 individuals, aged 22 to 45 years, are selected for the study. All 15 participants completed the study.

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28/34 [0051] The evaluation of the follicular biopsy species collected from the test sites indicated that the test and control materials generated an overall average evaluation score of 0.5 (1-24% very small hard masses). The average proportion of microcomponents per follicle is 1% for test and negative control materials, and 6% for positive control. There are no statistically significant differences in the proportions of microcomponents to follicles between the test and negative control materials, while the positive control demonstrated a significantly greater relationship than the negative control.

[0052] The study showed that there are no adverse experiences reported during the study period. The test materials did not show a potential to cause comedogenicity. The microcomponent ratios per follicle are not statistically different from the locations treated with each of the test materials and locations where an untreated negative control dressing is applied.

EXAMPLE 9 [0053] A study is conducted to determine the potential for skin irritation and sensitization of a test material in the form of eye cream, with the formulations described above. The tests are carried out during a study period of 8 weeks. This study started with 225 individuals. Eleven study subjects discontinued the study for reasons unrelated to the test material. A total of 214 patients completed the study.

[0054] Tests are performed by applying dressings with the test material to a test area that is cleaned with 70% isopropyl alcohol and left to dry. The test material is applied to the upper back (between the shoulder blades) and is allowed to remain in direct contact with the skin for a period of 24 hours. Dressings are applied to the same location on Monday, Wednesday and Friday for a total of nine applications during the Induction Period. This calendar

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29/34 may have been modified to allow missed visits or holidays. If a subject is unable to report on an assigned test date, the test material is applied on 2 consecutive days, during the Induction Phase and / or a composition day is added at the end of the induction phase. The sites are classified by a skin irritation supervision technician 24 hours after the removal of the dressings by the subjects on Tuesday and Thursday and 48 hours after the removal of the dressings on Saturday, unless the patch application schedule has been changed as described above.

[0055] Locations are classified according to the following scoring system:

Dermal Score Scale

No visible skin reaction ± Little noticeable erythema + Mild erythema + Well-defined erythema + Erythema and severe edema + Erythema and edema with vesiculation [0056] If a “2+” or greater reaction has occurred, the test material is applied to a site adjacent virgin. If a “2+” or greater reaction occurred at the new location, a dressing is not applied to the subject again during the Induction Phase, but is changed on the appropriate study day. At the discretion of the Study Director, the patch locations with scores below a “2+” may have been changed.

[0057] After approximately a 2-week rest period, the changed dressings are applied to previously untreated test sites on the back. After 24 hours, the patches are removed by a CRL technician and the test sites are evaluated for skin reactions. The test sites are reassessed at 48 and 72 hours. Subjects that exhibit

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30/34 reactions during the study's Phase of Change may have been asked to return for a 96-hour reading.

[0058] Based on a test population of 214 individuals and under the conditions of the present study, the test material, eye cream, has not shown a potential to induce skin irritation or sensitization.

EXAMPLE 10 [0059] A series of studies is conducted to determine the irritation and dermal sensitization potential of a test material in the form of day cream and eye cream, with the formulations described above, with additional sun protection factor (SPF) component added to formulations. SPF is defined as the ratio of the amount of energy required to produce minimal erythema on protected skin to the amount of energy required to produce minimal erythema on untreated skin calculated as follows:

SPF = Protected skin MED J / m ² MED

Unprotected Control Site J / m ² [0060] I. A study is conducted to evaluate the effectiveness of a sunscreen combined with a day cream, as formulated above by determining the sun protection factor (SPF) using the Test Method (May 2006) of the International Sun Protection Factor (SPF), under static conditions. A standard sunscreen formulation is tested simultaneously with the test material. The standard is Padimate O / Oxybenzone SPF Standard (COLIPA P2) (16.63 ± 3.43). The place of application of the test / control material is the back and it is at random that the area of the back for treatment with an individual test material is not the same for all individuals. A total of 10 male and female individuals, aged 25-57 years and, in general, in good health, are selected for the study. There are no adverse events reported during the course of the study.

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31/34 [0061] II. A study is conducted to evaluate the effectiveness of a sunscreen product with an SPF 30 combined with a day cream, as formulated above, determining the ultraviolet A (UVA) radiation protection factor (PFA), under static conditions, according to the Japanese Cosmetic Industrial Association (JCIA) UVA Protection Measurement Standard (01/01196). A total of 10 adult individuals participated in the study. The Xenon Arc Multi-Port Solar Simulator (150w, Model 15S, Solar Light Company, Philadelphia, PA) which has a spectrum of continuous ultraviolet light emission in the region of UVA and UVB radiation (290-400 nm), is used as a irradiation source of ultraviolet light and filtered to provide a spectrum of solar energy, base. The following filters are used to ensure adequate spectral results: Schott WG33513 mm and UG1111 mm. The lamp output is measured immediately before each subject's test begins with a UV intensity meter (Model PMA2100, Solar Light Company, Philadelphia, PA). The spectral distributions of the optical output of the solar simulators are validated annually.

[0062] Pigment darkening is a brown-gray to brown-black gray reaction seen in human skin after exposure to UVA rays, due to a photo-oxidation reaction in which a colorless melanin precursor is oxidized to become melanin pigmented. A Minimal Persistent Pigment Darkening (MPPD) is defined as the dose of UVA light irradiation required to induce such a response in untreated skin. Before the test phase, each subject's MPPD is determined by a progressive geometric sequence of UV light exposures. The increment of the UVA radiation dose is 1.25. Two to four hours after irradiation, sites are assessed for pigment browning according to the following scoring system:

Negative, Reaction not visible

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32/34 ± Little noticeable pigment darkening (minimum)

1+ Areolas distinct from unmistakable pigment darkening (moderate)

2+ Pronounced or well-defined pigment browning [0063] Test sites measuring 5 x 10 cm are described with a surgical marking pen on the subject's back between the shoulder blades and the waist line, lateral to the midline. These zones are designated for standard test material or sunscreen. An adjacent site is designated for a concurrent MPPD determination on untreated skin. After applying the product, each test area is subdivided into about 1-cm ² locations that are used for exposure to UVA light of the defined series. A 0.1 ml portion of test material or standard is applied to the appropriate test site and spread evenly over the site using a finger (equivalent to 2pl / cm ² ). Irradiation of the sites started between 15 minutes and 30 minutes after application. Exposure times are selected for each treated site based on the previously determined unprotected skin MPPD and the anticipated PFA of the test material or standard. All test sites are evaluated two to four hours after exposure to determine the minimum persistent pigment browning response.

[0064] A UVA radiation protection factor (PFA) is defined as the ratio of the amount of time required to produce an MPPD on protected skin to the amount of time required to produce an MPPD on untreated skin calculated as follows :

PFA = MPPD Protected Skin MPPD

Unprotected Control Location

Standard error must be within 10% of the measured values.

[0065] The entire value of the average PFA value of the treated irradiated sites determines the degree of protection from UVA. For all PFA values

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33/34 equivalent to 2 or higher, the following classification is applied to determine the degree of protection:

PFA PA value (Degree of protection of UVA) or more, but less than 4 PA + or more, but less than 8 PA + + or more PA + + + [0066] The Day Cream of the test material submitted with SPF 30 UVA exhibited an average PFA Protection Value of UVA of 6.5 in the 10 subjects.

[0067] III. A study is carried out to evaluate a day cream test material as formulated above, containing broad spectrum protection sunscreen actives, determining its critical wavelength in accordance with the FDA (Federal Register / Vol 76, No 117 / Friday, June 17, 2011 / Rules and Regulations).

[0068] The absorbance of a sunscreen is integrated (added) from 290 nm along the UV wavelengths until the sum reaches 90% of the total absorbance of the sunscreen in the ultraviolet region (290-400 nm ). The wavelength at which the combined absorption reaches 90% of the total absorbance is defined as the “critical wavelength” and is considered a measure of the range of sun protection. A sunscreen that has a significant part of its UVA absorption can be classified as "broad spectrum", when the critical wavelength is greater than 370 nm.

[0069] The spectral transmittance for each wavelength across the total UV spectrum (290 and 400 nm) is determined. Transmittance values are measured at 1 nanometer intervals.

[0070] The Critical Wavelength of the SPF day cream test material is 371.00 nm, which meets the minimum Wavelength criteria of 370 nm required for “broad spectrum” labeling.

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34/34 [0071] A similar study is conducted to evaluate a test of day cream material as formulated above, containing sunscreen actives for broad spectrum protection, determining in vitro the UVA protection factor (UVA-PF), its SPF / UVA-PF ratio and the critical wavelength of the day cream is in accordance with the COLIPA Guidelines, 2011.

[0072] Any patents or publications mentioned in this specification are hereby incorporated by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference.

[0073] The foregoing description is illustrative of the particular embodiments of the invention, but is not intended to be a limitation on its practice.

Claims (21)

1. Cosmetic in the form of an emulsion, the cosmetic characterized by the fact that it comprises: a fish roe protein isolate obtained from hatching fish eggs, wherein the isolate comprises an active protein that constitutes from 1 x 10 ^-8 to 1 x 10 ^-5 percent in total weight of the cosmetic; and a natural product extract comprising unsaturated fatty acids and sterols. 2. Cosmetic, according to claim 1, characterized by the fact that the natural product extract is an algae extract. 3. Cosmetic, according to claim 1, characterized by the fact that the active protein is zonase. Cosmetic according to any one of claims 1 to 3, characterized in that it additionally comprises 2-glucoside ascorbic acid or ascorbate salt. 5. Cosmetic, according to claim 1, characterized by the fact that the cosmetic is substantially free of silicone, petroleum jelly, synthetic paraben and mineral oil. 6. Cosmetic, according to claim 1, characterized by the fact that said fish roe protein isolate comprises active leukolectin. 7. Cosmetic according to any one of claims 1 to 6, characterized by the fact that fish eggs are salmon eggs. 8. Cosmetic according to any one of claims 1 to 6, characterized by the fact that fish eggs are eggs from at least one of white fish, coregono, cod, capelin and auction fish. 9. Cosmetic, characterized by the fact that it comprises: an hatching fish egg protein isolate obtained from hatching fish eggs comprising an active protein that Petition 870190052447, of 06/04/2019, p. 47/77 2/3 constitutes from 1 x 10 ^-8 to 1 x 10 ^-5 percent in total weight of the cosmetic; and at least one biocide. 10. Cosmetic, according to claim 9, characterized by the fact that the cosmetic is in the form of an emulsion. 11. Cosmetic, according to claim 10, characterized by the fact that the emulsion has an aqueous phase having a pH between 5.6 and 7.9. 12. Cosmetic, according to claim 10, characterized by the fact that it comprises Japanese honeysuckle extract. 13. Cosmetic according to claim 10, characterized by the fact that it comprises a natural product extract including unsaturated fatty acids and sterols. 14. Cosmetic, according to claim 13, characterized by the fact that the extract is derived from red algae, brown algae or tuft-forming cyanobacteria. 15. Cosmetic according to claim 10, characterized by the fact that the cosmetic additionally comprises ascorbic acid 2-glucoside or an ascorbate salt. 16. Cosmetic, according to claim 10, characterized by the fact that the active protein is zonase. 17. Cosmetic, according to claim 10, characterized by the fact that the active protein is leucolectin. 18. Cosmetic, according to claim 10, characterized by the fact that, on average, in a test population of human subjects, application of the cosmetic to the facial skin of each subject does not produce dermal irritation. 19. Cosmetic, according to claim 18, characterized by the fact that said active protein is zonase or leucolectin. Petition 870190052447, of 06/04/2019, p. 48/77 3/3 20. Kit, characterized by the fact that it comprises a cosmetic as defined in any one of claims 1 to 19 in a multipurpose package together with instructions for applying it to the skin to promote skin rejuvenation. 21. Process for improving the appearance of the skin, characterized by the fact that it comprises: application to a human's skin of a cosmetic as defined in any one of claims 1-8, 13, or 14 at a level of from 1 x 10 ^-6 to 0.1 mg of said fish roe protein isolate and 2 x 10 ^-7 to 0.02 mg of said unsaturated fatty acids per square centimeter of skin, at least three times a week to obtain an improvement in the appearance of the skin.