WO2008015342A2 - Use of a plant extract as active agent for increasing melanin synthesis in melanocytes - Google Patents

Use of a plant extract as active agent for increasing melanin synthesis in melanocytes Download PDF

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Publication number
WO2008015342A2
WO2008015342A2 PCT/FR2007/001334 FR2007001334W WO2008015342A2 WO 2008015342 A2 WO2008015342 A2 WO 2008015342A2 FR 2007001334 W FR2007001334 W FR 2007001334W WO 2008015342 A2 WO2008015342 A2 WO 2008015342A2
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WIPO (PCT)
Prior art keywords
extract
use according
skin
composition
hair
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Application number
PCT/FR2007/001334
Other languages
French (fr)
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WO2008015342A3 (en
Inventor
Claude Dal Farra
Nouha Domloge
Dominique Peyronel
Original Assignee
Societe D'extraction Des Principes Actifs Sa (Vincience)
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Filing date
Publication date
Priority claimed from FR0607096A external-priority patent/FR2904541B1/en
Priority claimed from FR0607095A external-priority patent/FR2904540B1/en
Priority claimed from FR0607093A external-priority patent/FR2904555B1/en
Priority claimed from FR0607101A external-priority patent/FR2904543B1/en
Priority claimed from FR0607106A external-priority patent/FR2904547B1/en
Application filed by Societe D'extraction Des Principes Actifs Sa (Vincience) filed Critical Societe D'extraction Des Principes Actifs Sa (Vincience)
Publication of WO2008015342A2 publication Critical patent/WO2008015342A2/en
Publication of WO2008015342A3 publication Critical patent/WO2008015342A3/en

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Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/55Linaceae (Flax family), e.g. Linum
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/28Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/31Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/48Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/88Liliopsida (monocotyledons)
    • A61K36/899Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9794Liliopsida [monocotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/18Antioxidants, e.g. antiradicals
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/04Preparations for care of the skin for chemically tanning the skin

Definitions

  • the present invention relates to the use of a plant extract in a cosmetic composition; the extract or composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb, in order to give it a tanned appearance, or to prepare the skin for exposure to the sun.
  • the present invention also relates to the use of a plant extract for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb for therapeutic purposes, for repigmenting depigmented skin, for example in the case of vitiligo, or for pigmenting hair or hair, especially in the treatment and prevention of canities.
  • the plant extract is mainly composed of protein and peptide compounds and is obtained from plants selected from the genera Linum, Zea, Helianthus and or species Glycine max and Brassica napus.
  • the color of human hair and skin is a function of various factors including seasons of the year, race, sex and age. It is mainly determined by the concentration of melanin produced by melanocytes. Melanocytes are specialized cells that, via particular organelles, melanosomes, synthesize melanin. The synthesis of melanin or melanogenesis is a complex process whose precise mechanisms are not yet elucidated and which schematically involves the following steps: Tyrosine * Dopa * " Dopaquinone * " Dopachrome * " Melanin
  • the melanocyte is involved in the epidermal melanic unit which comprises a melanocyte surrounded by about 40 neighboring keratinocytes.
  • melanin As melanin is synthesized in the melanosomes, they move from the perinuclear region to the end of the melanocyte dendrites.
  • phagocytosis By phagocytosis, the end of the dendrites is captured by keratinocytes, degraded membranes and redistributed melanosomes in the keratinocytes, where they will provide protection up to the natural desquamation of the cells.
  • the production of melanin, as well as its transport, are regulated by various factors such as, for example, UV radiation, hormones or chemicals.
  • an increase in exposure to UV radiation causes the synthesis of pigments and browning of the skin.
  • the pigmentation of hair and hair requires the presence of melanocytes in the bulb of the hair follicle.
  • the hair follicle is a tabular invagination of the epidemic that penetrates to the deep layers of the dermis.
  • the melanocytes in the bulb of the hair follicle are in an active state, that is, they synthesize melanin. These pigments are transmitted to the keratinocytes intended to form the hair shaft, which will lead to the growth of a pigmented hair or hair.
  • canitia naturally whitening of hair
  • melanin melanin
  • exogenous dye a dye, or a chemical substance, is meant to react with the superficial layers of the epidermis to cause the appearance of a pigmentation of the skin completely independent of the presence of melanin.
  • DHA dihydroxyacetone
  • vitiligo which is an autoimmune disease characterized by the appearance on the skin of white patches related to a lack of pigmentation or pityriasis versicolor, a superficial mycosis. causing the appearance of light spots that may appear immediately or after exposure to the sun.
  • the main object of the present invention is to provide a novel active agent which significantly promotes melanin synthesis in the melanocytes of the epidermis and the hair bulb.
  • the inventors have in fact demonstrated a biological activity and, more particularly dermatological and cosmetic, plant extracts obtained from plants selected from the genera Linum, Zea, Helianthus and or species Glycine max and Brassica napus.
  • the use of plant extracts obtained from this type of plant in the field of cosmetics has already been described in numerous patents, such as patent FR 2,698,785, which describes a composition for regulating the melanocyte system based on the fractions.
  • pro-pigmenting active agent a compound capable of promoting the increase of melanin synthesis in the melanocytes of the epidemic or hair bulb.
  • hair and hair we mean all the hairy appendages and in particular also eyelashes and eyebrows.
  • compositions according to the invention may be applied by any appropriate route, in particular oral, parenteral or external topical, and their formulation will be adapted by those skilled in the art, in particular for cosmetic or dermatological compositions.
  • the compositions according to the invention are intended for topical administration. They contain a physiologically acceptable medium, in particular a cosmetologically or pharmaceutically acceptable medium, in particular a dermatologically acceptable medium.
  • the subject of the invention is primarily the use in a cosmetic composition of a plant extract, consisting mainly of compounds of protein and peptide nature; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb.
  • extract refers to any isolated substance or preparation obtained from plant material. It is obtained, for example, by dissolving the active components by means of solvents (such as water, alcohol, etc.) then by concentration and purification of these active ingredients, for example by evaporation of the solvents.
  • solvents such as water, alcohol, etc.
  • one of the many plants of the genera Linum, Zea, Helianthus or species Glycine max and Brassica napus is used.
  • the plants of the genus Helianthus the species Helianthus annuus L. (sunflower) can advantageously be used according to the invention.
  • Linum plants it is advantageous to use, according to the invention, the species Linum usitatissimum L.
  • the Zea mays L. species can advantageously be used according to the invention.
  • the plant material used will be the grain and preferentially the grain freed from its envelope by a dehulling step. Any method of extraction or purification known to those skilled in the art can be used to prepare the hydrolyzate according to the invention.
  • the plant is ground using a plant mill.
  • the powder thus obtained may subsequently be "delipidated” with the aid of a conventional organic solvent (such as, for example, an alcohol, hexane or acetone).
  • a residue enriched in proteins is obtained, commonly called “cake”.
  • a dephenolization step can then be performed.
  • dephenolized is meant a product that contains less than 0.005% phenols and polyphenols (eg, chlorogenic acid, caffeic acid and quinic acid).
  • polyphenols eg, chlorogenic acid, caffeic acid and quinic acid.
  • the step of the process corresponding to the dephenolization is important insofar as the polyphenols are undesirable compounds in the cosmetic compositions. Indeed, they have the disadvantage of binding to proteins, which gives them a tanning effect, proteins becoming thereby insoluble in water.
  • polyphenols are colored and evolve over time, which causes an unsightly effect of the compositions, including creams, which contain them.
  • the extraction of proteins from the plant is then conventionally carried out (Osborne, 1924); the crushed plant being suspended in an alkaline solution.
  • the soluble fraction is collected after centrifugation and filtration steps, this crude solution then constituting a first form of the extract containing the proteins, carbohydrates and optionally lipids.
  • Proteins are then precipitated by varying the ionic strength and acidifying the medium to remove soluble components and nucleic acids.
  • the precipitate is then washed with a solvent such as, for example, ethanol or methanol and the solvent is evaporated by drying in vacuo.
  • the protein-rich precipitate is redissolved in water or other solvent and then constitutes a more purified form of the extract.
  • the extraction can also be carried out in neutral or acidic medium.
  • the precipitation step is then carried out using a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate).
  • the precipitate obtained can be separated from the precipitating agents by dialysis after redissolving in water or another solvent. At this stage, the extract obtained contains at least 70% of compounds of protein and peptide nature.
  • the isolated protein fraction according to the invention is then hydrolyzed under mild conditions to generate polypeptides and soluble peptides.
  • Hydrolysis is defined as a chemical reaction involving the cleavage of a molecule by water, this reaction being possible in a neutral, acidic or basic medium.
  • the hydrolysis is carried out chemically and / or advantageously by proteolytic enzymes.
  • endoproteases and exopeptidases of plant origin papain, bromelain, ficin
  • papain, bromelain, ficin originating from microorganisms may be mentioned. (Aspergillus, Rhizopus, Bacillus, etc.), or by so-called "industrial” enzymes such as alkalases, flavourzyme, etc.
  • the culture medium thus hydrolyzed is centrifuged and filtered until a protein hydrolyzate is obtained.
  • the solution obtained constitutes the active extract.
  • the active extract may be further purified by fractionation, in particular by a chromatographic or purified method and concentrated by a dialysis method. Any of the more or less purified forms of the extract is then solubilized in water or in any mixture containing water, and then sterilized by ultrafiltration.
  • the plant extract obtained according to the invention is analyzed qualitatively and quantitatively for its content of compounds of protein and peptide nature. Protein and peptide compounds are understood to mean protein fragments, peptides and free amino acids present in the mixture.
  • the active plant extract contains a quantity of protein and peptide compounds representing between 30 and 90% of the total weight of the dry matter, more particularly this amount is between 60 and 90% of the total weight of the dry matter. and 80% of the total weight of the dry matter, for extracts of soybean, corn and sunflower, or more particularly that amount is between 50 to 80% for extracts of flax and rapeseed.
  • the subject of the invention is the use of a plant extract according to the invention for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or of the hair bulb in order to repigment the depigmented skin, for example in the case of vitiligo, or in order to pigment hair or hair , especially in the treatment and prevention of canities.
  • the invention also relates to the use of a plant extract according to the invention for the preparation of a pharmaceutical composition; the extract or the composition being intended to attenuate the pigmentary disorders associated with skin hypopigmentation, in particular vitiligo.
  • the invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended to promote the pigmentation of the hairs and / or the hair.
  • the invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended to prepare the skin for exposure to the sun.
  • the invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended, by increasing melanin synthesis, to protect the skin from sunlight.
  • an extract according to the invention or the composition containing it is intended to protect the skin from environmental stresses.
  • the extract is first solubilized in one or more cosmetically or pharmaceutically acceptable solvents such as water, glycerol, propylene glycol, butylene glycol, ethoxylated or propoxylated diglycols, ethanol, propanol or isopropanol.
  • one or more cosmetically or pharmaceutically acceptable solvents such as water, glycerol, propylene glycol, butylene glycol, ethoxylated or propoxylated diglycols, ethanol, propanol or isopropanol.
  • the extract is first solubilized in a cosmetic or pharmaceutical vector such as liposomes or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites, and more generally solubilized in or attached to any cosmetically or pharmaceutically acceptable carrier.
  • a cosmetic or pharmaceutical vector such as liposomes or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites, and more generally solubilized in or attached to any cosmetically or pharmaceutically acceptable carrier.
  • the sunflower and corn extracts are used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in one amount representing from 0.05% to 10% of the total weight of the composition.
  • the flax, soya and rapeseed extracts are used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing from 0.05% to 5% of the total weight of the composition.
  • the content of protein and peptide compounds in the compositions of the present invention will then be from 0.00003% to 18% and preferably from 0.015% to 9% of the total weight of the composition, for plant extracts of the present invention.
  • sunflower and corn containing between 30 and 90% of compounds of protein and peptide nature.
  • the content of protein and peptide compounds in the compositions of the present invention will then be from 0.0006% to 16% and preferably from 0.03% to 8% of the total weight of the composition, for plant extracts of sunflower and corn, containing between 60 and 80% of compounds of protein and peptide nature.
  • the content of protein and peptide compounds in the compositions of the present invention will then be from 0.00003% to 18% and preferably from 0.015% to 9% of the total weight of the composition, for plant extracts of flax, soybean and rapeseed, containing between 30 and 90% of compounds of protein and peptide nature.
  • the content of protein and peptide compounds in the compositions of the present invention will then be from 0.0006% to 16% and preferably from 0.03% to 4% of the total weight of the composition, for plant extracts of soya , containing between 60 and 80% of compounds of protein and peptide nature.
  • the content of protein and peptide compounds in the compositions of the present invention will then be from 0.0005% to 16% and preferably from 0.025% to 4% of the total weight of the composition, for plant extracts of flax and rapeseed, containing between 50 and 80% of compounds of protein and peptide nature.
  • a pro-pigmenting compound according to the invention offers outlets in self-tanning products and products promoting tanning.
  • the invention relates to the use of extract for the manufacture of products for artificial tanning and / or browning of the skin.
  • the invention also relates to the use of extract to improve the intensity and / or the homogeneity and / or the persistence of the coloration produced on the skin and / or the hair.
  • compositions according to the present invention will be in a form suitable for topical administration to the skin, and cover all cosmetic or dermatological forms.
  • These compositions must therefore contain a medium cosmetically and / or dermatologically acceptable, that is to say compatible with the skin, hair or hair.
  • These compositions may especially be in the form of creams, O / W emulsions, W / O emulsions or multiple emulsions, solutions, suspensions, gels, milks, lotions, sticks or even powders, suitable for application to the skin, lips and / or the hair.
  • compositions comprise the excipients necessary for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes.
  • the composition that may be used according to the invention may in particular consist of a composition for hair care, and in particular a shampoo, a conditioner, a setting lotion, a treatment lotion, a cream or a styling gel, a restructuring lotion for hair, a mask, etc.
  • the cosmetic composition according to the invention can be used in particular in treatments using an application that is followed or not followed by rinsing, or in the form of shampoo. It can also be in the form of dye or mascara to be applied with a brush or a comb, in particular on eyelashes, eyebrows or hair.
  • plant extracts mainly consisting of protein and peptide compounds, according to the invention may be used as the only pro-pigmenting agent or in combination with at least one other active agent, in a composition cosmetic or for the preparation of a dermatological and / or pharmaceutical composition.
  • compositions that can be used according to the invention also contain at least one other agent that promotes the pigmentation of the skin, hair and / or hair.
  • agents are in particular tyrosinase substrates, such as tyrosine or L-DOPA, prostaglandins, or activating compounds of the cAMP pathway such as pro-opiomelanocortin derivatives, adenosine, or forkoline or its derivatives.
  • Plant extracts such as bitter orange (citrus aurantium) or chrysanthemum (Chrysanthemum morifoliurri), described in particular in patents FR 2845285 and EP1014934.
  • Such compounds are also found in the family of exogenous dyes of the superficial layers of the epidermis, such as dihydroxyacetone (DHA), perythrulose, extracts of henna leaves, described in particular in patents EP 0742002 and FR2779958.
  • DHA dihydroxyacetone
  • perythrulose perythrulose
  • extracts of henna leaves described in particular in patents EP 0742002 and FR2779958.
  • Example 1 Preparation of a Linseed Extract (Linum usitatissimum L.)
  • 1 kg of flaxseed is delipidated by the action of an organic solvent: hexane.
  • a residue enriched in proteins (the cake) is obtained.
  • the residue is then diluted in a slightly basic aqueous solution and then hydrolyzed by adding an enzyme: papain, at 60 ° C. for 4 hours with stirring.
  • This mixture is then heated to 80 ° C, centrifuged and filtered until a clear solution is obtained. It is then concentrated under vacuum, then filtered again on filter plates and sterilizing cartridge.
  • a hydrolyzate titrating at 30 g / l of compounds of protein nature is then obtained.
  • the hydrolyzed extract contains a quantity of protein compounds representing approximately 65% of the total weight of the dry matter. This hydrolyzate is redissolved in dipropylene glycol.
  • Example 2 Preparation of Dephenolated Extracts from Corn (Zea Mays) 100 grams of maize cake are initially dephenolized to a phenol content of less than 0.005%.
  • the product is enzymatically hydrolyzed in aqueous medium by an enzymatic cocktail (protease, lipase and cellulase) at different pH and temperatures for 4 to 8 hours and then filtered.
  • Activated carbons are then used to obtain a product that satisfies the cosmetic criteria in terms of odor and color.
  • the product is finally sterilized by sterilizing filtration and heating.
  • the hydrolyzate thus obtained contains more than 80% of proteins and less than 0.005% of phenols.
  • EXAMPLE 3 Preparation of Dephenolated Extracts from Sunflower (Helianthus annuus) 100 grams of sunflower cake are initially dephenolized until a phenol content of less than 0.005% is obtained. Then the proteins are precipitated in order to increase their concentration. Then, the product is enzymatically hydrolyzed in aqueous medium by an enzymatic cocktail (protease, lipase and cellulase) at different pH and temperatures for 4 to 8 hours and then filtered. Activated carbons are then used to obtain a product that satisfies the cosmetic criteria in terms of odor and color. The product is finally sterilized by sterilizing filtration and heating. The hydrolyzate thus obtained contains more than 80% of proteins and less than 0.005% of phenols.
  • the cakes are the solid residues obtained after extraction of the oil from the soya beans. They represent 50 to 75% of the seed mass.
  • the cakes are suspended in an aqueous alkaline solution (1/10 dilution). This mixture is stirred for a time long enough to allow the solubilization of the soluble fractions.
  • the extraction temperature is variable (between 4 and 80 ° C); preferably the operation will be performed cold.
  • the medium is clarified by centrifugation and then filtered through a plate filter.
  • This filtrate which contains the soluble fractions of soybeans, is then precipitated by varying the ionic strength in a neutral or acidic medium to remove soluble carbohydrate components, lipids, and nucleic acids.
  • the supernatant is removed and the precipitate is then washed with a solvent such as, for example, ethanol or methanol and the solvent is evaporated by drying in vacuo.
  • the protein-rich precipitate is dissolved in water or another solvent.
  • the extraction can also be carried out in neutral or acidic medium.
  • the crude protein extract is then subjected to a series of gentle and selective hydrolyses consisting of chemical and enzymatic hydrolyses.
  • hydrolysis in the presence of exopeptidases (papain, ficin).
  • the hydrolyzate is then purified and membrane-dialyzed to remove the high molecular weight compounds.
  • a light-colored hydrolyzate titrant of 15 to 30 g / l of dry extract and containing a quantity of compounds of protein and peptide nature representing approximately 70% of the total weight of the dry matter is then obtained.
  • the hydrolyzate is then diluted so that the protein concentration determined by the Lowry method is between 0.5 and 2 g / l.
  • Example 5 Preparation of an extract of rapeseed (Brassica napus).
  • a first step 1 kg of the rapeseeds are delipidated by the action of an organic solvent: hexane. After drying the product, a residue enriched in proteins (the cake) is obtained. The cake is then diluted in a slightly basic aqueous solution and then hydrolyzed by adding an enzyme: papain, at 60 ° C. for 4 hours with stirring. This mixture is then heated to 80 ° C., centrifuged and filtered until a clear solution is obtained. It is then concentrated under vacuum, then filtered again on filter plates and sterilizing cartridge. A brown-colored hydrolyzate titrating at 30 g / l of protein-like compounds is then obtained. The hydrolyzed extract contains a quantity of protein compounds representing approximately 65% of the total weight of the dry matter. This hydrolyzate is redissolved in dipropylene glycol.
  • compositions were obtained by simple mixing of the various components. The quantities indicated are given as a percentage of weight.
  • Aqueous phase Aqueous phase :

Abstract

The present invention relates to the use of a plant extract, alone or in combination with at least one other active agent, in a cosmetic composition or for the preparation of a pharmaceutical composition, as active agent intended to promote increased melanin synthesis in the melanocytes of the epidermis or of the hair bulb. The plant extract is mainly constituted of compounds of protein and peptide nature and is obtained from plants selected from the genera Linum, Zea and Helianthus or the species Glycine max and Brassica napus.

Description

UTILISATION D'UN EXTRAIT VEGETAL EN TANT QU'AGENT ACTIF POUR AUGMENTER LA SYNTHESE DE MELANINE DANS LES MELANOCYTES USE OF PLANT EXTRACT AS ACTIVE AGENT TO INCREASE MELANIN SYNTHESIS IN MELANOCYTES
La présente invention concerne l'utilisation d'un extrait végétal dans une composition cosmétique ; l'extrait ou la composition étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épiderme ou du bulbe pileux, en vue de lui donner un aspect bronzé, ou de préparer la peau à une exposition au soleil. La présente invention concerne également l'utilisation d'un extrait végétal pour la préparation d'une composition pharmaceutique ; l'extrait ou la composition étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épiderme ou du bulbe pileux à des fins thérapeutiques, pour repigmenter la peau dépigmentée par exemple dans le cas du vitiligo, ou pour pigmenter les poils ou les cheveux, en particulier dans le traitement et la prévention de la canitie. L'extrait végétal est principalement constitué de composés de nature protéique et peptidique et est obtenu à partir de plantes choisies parmi les genres Linum, Zea, Helianthus et ou des espèces Glycine max et Brassica napus.The present invention relates to the use of a plant extract in a cosmetic composition; the extract or composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb, in order to give it a tanned appearance, or to prepare the skin for exposure to the sun. The present invention also relates to the use of a plant extract for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb for therapeutic purposes, for repigmenting depigmented skin, for example in the case of vitiligo, or for pigmenting hair or hair, especially in the treatment and prevention of canities. The plant extract is mainly composed of protein and peptide compounds and is obtained from plants selected from the genera Linum, Zea, Helianthus and or species Glycine max and Brassica napus.
La couleur des cheveux et de la peau humaine est fonction de différents facteurs et notamment des saisons de l'année, de la race, du sexe et de l'âge. Elle est principalement déterminée par la concentration de mélanine produite par les mélanocytes. Les mélanocytes sont les cellules spécialisées qui, par l'intermédiaire d'organites particuliers, les mélanosomes, synthétisent la mélanine. La synthèse de la mélanine ou mélanogénèse est un processus complexe dont les mécanismes précis ne sont pas encore élucidés et qui fait intervenir schématiquement les étapes suivantes : Tyrosine * Dopa *" Dopaquinone *" Dopachrome *" MélanineThe color of human hair and skin is a function of various factors including seasons of the year, race, sex and age. It is mainly determined by the concentration of melanin produced by melanocytes. Melanocytes are specialized cells that, via particular organelles, melanosomes, synthesize melanin. The synthesis of melanin or melanogenesis is a complex process whose precise mechanisms are not yet elucidated and which schematically involves the following steps: Tyrosine * Dopa * " Dopaquinone * " Dopachrome * " Melanin
Dans l'épiderme, le mélanocyte est impliqué dans l'unité mélanique épidermique qui comporte un mélanocyte entouré d'environ 40 kératinocytes voisins. Au fur et à mesure que la mélanine est synthétisée dans les mélanosomes, ceux-ci se déplacent de la région périnucléaire vers l'extrémité des dendrites des mélanocytes. Par phagocytose, l'extrémité des dendrites est capturée par les kératinocytes, les membranes dégradées et les mélanosomes redistribués dans les kératinocytes, où ils assureront une protection jusqu'à la desquamation naturelle des cellules. La production de mélanine, ainsi que son transport, sont régulés par différents facteurs tels que, par exemple, le rayonnement UV, les hormones ou les produits chimiques. Ainsi, une augmentation de l'exposition aux rayonnements UV provoque la synthèse de pigments et le brunissement de la peau. La pigmentation du cheveu et des poils requiert la présence de mélanocytes au niveau du bulbe du follicule pileux. Le follicule pileux est une invagination tabulaire de l'épidémie qui s'enfonce jusqu'aux couches profondes du derme. Les mélanocytes au niveau du bulbe du follicule pileux sont dans un état actif, c'est-à-dire qu'ils synthétisent de la mélanine. Ces pigments sont transmis aux kératinocytes destinés à former la tige pilaire, ce qui conduira à la pousse d'un cheveu ou d'un poil pigmenté.In the epidermis, the melanocyte is involved in the epidermal melanic unit which comprises a melanocyte surrounded by about 40 neighboring keratinocytes. As melanin is synthesized in the melanosomes, they move from the perinuclear region to the end of the melanocyte dendrites. By phagocytosis, the end of the dendrites is captured by keratinocytes, degraded membranes and redistributed melanosomes in the keratinocytes, where they will provide protection up to the natural desquamation of the cells. The production of melanin, as well as its transport, are regulated by various factors such as, for example, UV radiation, hormones or chemicals. Thus, an increase in exposure to UV radiation causes the synthesis of pigments and browning of the skin. The pigmentation of hair and hair requires the presence of melanocytes in the bulb of the hair follicle. The hair follicle is a tabular invagination of the epidemic that penetrates to the deep layers of the dermis. The melanocytes in the bulb of the hair follicle are in an active state, that is, they synthesize melanin. These pigments are transmitted to the keratinocytes intended to form the hair shaft, which will lead to the growth of a pigmented hair or hair.
Il est admis que la canitie (blanchiment naturel des cheveux) est associée à une diminution de mélanine dans la tige pilaire.It is accepted that canitia (natural whitening of hair) is associated with a decrease of melanin in the hair shaft.
De nos jours, il est important d'avoir bonne mine et une peau bronzée est toujours signe de bonne santé. Cependant, le bronzage naturel n'est pas toujours sans risque dans la mesure où il nécessite des expositions prolongées aux rayonnements UV, qui sont susceptibles en contrepartie d'induire une altération des cellules cutanées, notamment dans le cas des individus de phototypes clairs. La peau vieillit prématurément, devient sèche et se caractérise par de nombreuses rides et taches de vieillesse. Il est donc souhaitable de trouver une alternative au bronzage naturel qui soit compatible avec les exigences de santé des individus. D'autre part, on sait que dans la plupart des populations le maintien d'une coloration constante de la chevelure, signe de jeunesse, est une aspiration forte.Nowadays, it is important to look good and tanned skin is always a sign of good health. However, natural tanning is not always without risk insofar as it requires prolonged exposures to UV radiation, which are likely in return to induce an alteration of skin cells, especially in the case of individuals of clear phototypes. The skin ages prematurely, becomes dry and is characterized by many wrinkles and age spots. It is therefore desirable to find an alternative to natural tanning that is compatible with the health requirements of individuals. On the other hand, we know that in most populations the maintenance of a constant coloring of the hair, sign of youth, is a strong aspiration.
La recherche de composés pouvant favoriser la synthèse de la mélanine dans la peau et les cheveux est une préoccupation de la dermatologie et de la cosmétique. Ces nouveaux composés seraient notamment utiles comme alternative aux expositions solaires, pour préparer la peau et protéger celle-ci des rayons du soleil, pour obtenir un bronzage plus intense après une exposition au soleil, pour prolonger la pigmentation naturelle de la peau après une exposition au soleil ou pour prévenir et/ou limiter et/ou stopper le développement de la canitie et même maintenir la pigmentation naturelle des cheveux et/ou des poils gris ou blancs. On entend par pigmentation naturelle de la peau, la coloration de la peau ou des cheveux déterminée par la concentration en mélanine.The search for compounds that can promote the synthesis of melanin in the skin and hair is a concern of dermatology and cosmetics. These new compounds are particularly useful as an alternative to sun exposure, to prepare the skin and protect it from the sun's rays, to obtain a tan more intense after exposure to the sun, to prolong the natural pigmentation of the skin after exposure to the sun. sun or to prevent and / or limit and / or stop the development of canities and even maintain the natural pigmentation of hair and / or gray or white hairs. By natural pigmentation of the skin is meant the coloring of the skin or hair determined by the melanin concentration.
A cet égard, il a été proposé de nombreuses solutions par apport de colorants exogènes. On entend par colorant exogène, un colorant, ou une substance chimique, capable de réagir avec les couches superficielles de Fépiderme pour provoquer l'apparition d'une pigmentation de la peau tout à fait indépendante de la présence de mélanine. L'un des colorants exogènes le plus connu est la dihydroxyacétone, ou DHA. Toutefois, seule la stimulation de la pigmentation de la peau et/ou des cheveux par la voie naturelle permet une réelle protection vis à vis des rayonnements UV et reste la voie idéale de stimulation de la pigmentation. Ainsi il a été proposé dans l'art antérieur la préparation et l'utilisation d'activateurs de la biosynthèse de la mélanine (FR 2828097, FR 2831438, FR 2845285) faisant intervenir des hormones (alpha MSH ou ses dérivés WO2006037188) ou des prostaglandines (WO9511003).In this respect, many solutions have been proposed by adding exogenous dyes. By exogenous dye, a dye, or a chemical substance, is meant to react with the superficial layers of the epidermis to cause the appearance of a pigmentation of the skin completely independent of the presence of melanin. One of the best known exogenous dyes is dihydroxyacetone, or DHA. However, only the stimulation of the pigmentation of the skin and / or the hair by the natural way allows a real protection vis-à-vis the UV radiations and remains the ideal way of stimulation of the pigmentation. Thus, it has been proposed in the prior art the preparation and use of activators for the biosynthesis of melanin (FR 2828097, FR 2831438, FR 2845285) involving hormones (alpha MSH or its derivatives WO2006037188) or prostaglandins. (WO9511003).
D'autre part, il existe des pathologies liées à la pigmentation comme le vitiligo qui est une maladie auto-immune se caractérisant par l'apparition sur la peau de plaques blanches liées à un déficit de pigmentation ou encore le pityriasis versicolor, une mycose superficielle provoquant l'apparition de taches claires pouvant se manifester d'emblée ou après une exposition au soleil.On the other hand, there are pathologies related to pigmentation such as vitiligo which is an autoimmune disease characterized by the appearance on the skin of white patches related to a lack of pigmentation or pityriasis versicolor, a superficial mycosis. causing the appearance of light spots that may appear immediately or after exposure to the sun.
La présente invention a pour principal objectif de fournir un nouvel agent actif favorisant de façon importante la synthèse de mélanine dans les mélanocytes de Pépiderme et du bulbe pileux. Les inventeurs ont en effet mis en évidence une activité biologique et, plus particulièrement dermatologique et cosmétique, d'extraits végétaux obtenus à partir de plantes choisies parmi les genres Linum, Zea, Helianthus et ou des espèces Glycine max et Brassica napus. L'utilisation d'extraits végétaux obtenus à partir de ce type de plantes dans le domaine de la cosmétique a déjà été décrite dans de nombreux brevets, tels que le brevet FR 2 698 785 qui décrit une composition régulatrice du système mélanocytaire à base des fractions insaponifiables du maïs, ou le brevet WO 2004/01096 décrivant des compositions à base de polyphénols de lin, ou encore le brevet US 5 698 184 5, décrivant un actif pigmentant à base de peptones de soja complexées à des ions métalliques. Toutefois, à la connaissance de la demanderesse, l'utilisation d'extraits végétaux, principalement constitués de composés de nature protéique et peptidique, obtenus à partir de plantes choisies parmi les genres Linum, Zea, Helianthus ou des espèces Glycine max et Brassica napus, pour favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épidémie ou du bulbe pileux, n'a pas encore été exposée.The main object of the present invention is to provide a novel active agent which significantly promotes melanin synthesis in the melanocytes of the epidermis and the hair bulb. The inventors have in fact demonstrated a biological activity and, more particularly dermatological and cosmetic, plant extracts obtained from plants selected from the genera Linum, Zea, Helianthus and or species Glycine max and Brassica napus. The use of plant extracts obtained from this type of plant in the field of cosmetics has already been described in numerous patents, such as patent FR 2,698,785, which describes a composition for regulating the melanocyte system based on the fractions. unsaponifiable maize, or patent WO 2004/01096 describing compositions based on linolenic polyphenols, or patent US 5,698,184 5, describing a pigmenting agent based on soya peptones complexed with metal ions. However, to the knowledge of the applicant, the use of plant extracts, mainly consisting of protein and peptide compounds, obtained from plants selected from the genera Linum, Zea, Helianthus or species Glycine max and Brassica napus, to promote the increase of melanin synthesis in the melanocytes of the epidemic or hair bulb, has not yet been exposed.
Par agent actif pro-pigmentant, on entend un composé capable de favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épidémie ou du bulbe pileux.By pro-pigmenting active agent is meant a compound capable of promoting the increase of melanin synthesis in the melanocytes of the epidemic or hair bulb.
Par poils et cheveux on entend l'ensemble des annexes pileuses et notamment également les cils et les sourcils.By hair and hair we mean all the hairy appendages and in particular also eyelashes and eyebrows.
Les compositions selon l'invention pourront être appliquées par toute voie appropriée, notamment orale, parentérale ou topique externe, et leur formulation sera adaptée par l'homme du métier, en particulier pour des compositions cosmétiques ou dermatologiques. Avantageusement, les compositions selon l'invention sont destinées à une administration par voie topique. Elles contiennent un milieu physiologiquement acceptable, en particulier un milieu cosmétologiquement ou pharmaceutiquement, notamment dermatologiquement acceptable.The compositions according to the invention may be applied by any appropriate route, in particular oral, parenteral or external topical, and their formulation will be adapted by those skilled in the art, in particular for cosmetic or dermatological compositions. Advantageously, the compositions according to the invention are intended for topical administration. They contain a physiologically acceptable medium, in particular a cosmetologically or pharmaceutically acceptable medium, in particular a dermatologically acceptable medium.
Ainsi, l'invention a pour objet premier l'utilisation dans une composition cosmétique d'un extrait végétal, principalement constitué de composés de nature protéique et peptidique ; l'extrait ou la composition étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de Fépiderme ou du bulbe pileux.Thus, the subject of the invention is primarily the use in a cosmetic composition of a plant extract, consisting mainly of compounds of protein and peptide nature; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb.
Le terme "extrait" désigne toute substance ou préparation isolée, obtenue à partir de matière végétale. Il s'obtient, par exemple, par dissolution des composants actifs au moyen de solvants (comme l'eau, l'alcool, etc.) puis par concentration et purification de ces actifs, par exemple, par évaporation des solvants.The term "extract" refers to any isolated substance or preparation obtained from plant material. It is obtained, for example, by dissolving the active components by means of solvents (such as water, alcohol, etc.) then by concentration and purification of these active ingredients, for example by evaporation of the solvents.
Plus particulièrement selon l'invention on utilise une des nombreuses plantes des genres Linum, Zea, Helianthus ou des espèces Glycine max et Brassica napus. Lorsque l'on utilise les plantes du genre Helianthus on peut utiliser, avantageusement selon l'invention, l'espèce Helianthus annuus L. (tournesol). Lorsque l'on utilise les plantes du genre Linum, on peut utiliser, avantageusement selon l'invention, l'espèce Linum usitatissimum L.More particularly according to the invention, one of the many plants of the genera Linum, Zea, Helianthus or species Glycine max and Brassica napus is used. When using the plants of the genus Helianthus, the species Helianthus annuus L. (sunflower) can advantageously be used according to the invention. When Linum plants are used, it is advantageous to use, according to the invention, the species Linum usitatissimum L.
Lorsque l'on utilise les plantes du genre Zea on peut utiliser, avantageusement selon l'invention, l'espèce Zea mays L.When plants of the genus Zea are used, the Zea mays L. species can advantageously be used according to the invention.
Selon l'invention le matériel végétal utilisé sera le grain et préférentiellement le grain débarrassé de son enveloppe par une étape de décorticage. Toute méthode d'extraction ou de purification connue de l'homme du métier peut être utilisée afin de préparer l'hydrolysat selon l'invention.According to the invention, the plant material used will be the grain and preferentially the grain freed from its envelope by a dehulling step. Any method of extraction or purification known to those skilled in the art can be used to prepare the hydrolyzate according to the invention.
Plus particulièrement, dans une première étape, la plante est broyée à l'aide d'un broyeur à plantes. La poudre ainsi obtenue peut ultérieurement être "délipidée" à l'aide d'un solvant organique classique (comme par exemple un alcool, un hexane ou de l'acétone). Après séchage du produit ainsi obtenu, on obtient un résidu enrichi en protéines communément appelé « tourteau ».More particularly, in a first step, the plant is ground using a plant mill. The powder thus obtained may subsequently be "delipidated" with the aid of a conventional organic solvent (such as, for example, an alcohol, hexane or acetone). After drying of the product thus obtained, a residue enriched in proteins is obtained, commonly called "cake".
Une étape de déphénolisation peut ensuite être réalisée. On entend par le terme «déphénolisé» un produit qui contient moins de 0,005 % de phénols et de polyphénols (par exemple, l'acide chlorogénique, l'acide caféique et l'acide quinique). L'étape du procédé correspondant à la déphénolisation est importante dans la mesure où les polyphénols sont des composés indésirables dans les compositions cosmétiques. En effet, ils ont comme inconvénient de se lier aux protéines, ce qui leur confère un effet tannant, les protéines devenant par là même insolubles dans l'eau. Par ailleurs, les polyphénols sont colorés et évoluent dans le temps, ce qui provoque un effet inesthétique des compositions, notamment des crèmes, qui les contiennent.A dephenolization step can then be performed. By the term "dephenolized" is meant a product that contains less than 0.005% phenols and polyphenols (eg, chlorogenic acid, caffeic acid and quinic acid). The step of the process corresponding to the dephenolization is important insofar as the polyphenols are undesirable compounds in the cosmetic compositions. Indeed, they have the disadvantage of binding to proteins, which gives them a tanning effect, proteins becoming thereby insoluble in water. Furthermore, polyphenols are colored and evolve over time, which causes an unsightly effect of the compositions, including creams, which contain them.
On réalise ensuite classiquement l'extraction des protéines de la plante (Osborne, 1924) ; le broyât de plante étant mis en suspension dans une solution alcaline. La fraction soluble est recueillie après des étapes de centrifugation et de filtration, cette solution brute constituant alors une première forme de l'extrait contenant les protéines, les glucides et éventuellement des lipides. Les protéines sont ensuite précipitées en faisant varier la force ionique et en acidifiant le milieu, ce qui permet d'éliminer les composants solubles et les acides nucléiques. Le précipité est ensuite lavé à l'aide d'un solvant tel que, par exemple, l'éthanol ou le méthanol puis le solvant est évaporé par séchage sous vide. Le précipité riche en protéines est remis en solution dans l'eau ou un autre solvant et constitue alors une forme plus purifiée de l'extrait.The extraction of proteins from the plant is then conventionally carried out (Osborne, 1924); the crushed plant being suspended in an alkaline solution. The soluble fraction is collected after centrifugation and filtration steps, this crude solution then constituting a first form of the extract containing the proteins, carbohydrates and optionally lipids. Proteins are then precipitated by varying the ionic strength and acidifying the medium to remove soluble components and nucleic acids. The precipitate is then washed with a solvent such as, for example, ethanol or methanol and the solvent is evaporated by drying in vacuo. The protein-rich precipitate is redissolved in water or other solvent and then constitutes a more purified form of the extract.
L'extraction peut également être réalisée en milieu neutre ou acide. L'étape de précipitation s'effectue alors à l'aide d'un agent classique de précipitation tel que les sels (chlorure de sodium, sulfate d'ammonium). Le précipité obtenu peut être séparé des agents de précipitation par dialyse après remise en solution dans de l'eau ou un autre solvant. A ce stade, l'extrait obtenu contient au minimum 70 % de composés de nature protéique et peptidique.The extraction can also be carried out in neutral or acidic medium. The precipitation step is then carried out using a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate). The precipitate obtained can be separated from the precipitating agents by dialysis after redissolving in water or another solvent. At this stage, the extract obtained contains at least 70% of compounds of protein and peptide nature.
La fraction protéique isolée selon l'invention est ensuite hydrolysée dans des conditions ménagées pour générer des polypeptides et des peptides solubles. L'hydrolyse se définit comme étant une réaction chimique impliquant le clivage d'une molécule par de l'eau, cette réaction pouvant se faire en milieu neutre, acide ou basique. Selon l'invention, l'hydrolyse est réalisée par voie chimique et/ou de façon avantageuse par des enzymes protéolytiques. On peut alors citer l'utilisation des endoprotéases et exopeptidases d'origine végétale (papaïne, bromelaïhe, ficine), provenant de micro-organismes (Aspergillus, Rhizopus, Bacillus, etc.), ou bien par des enzymes dites « industrielles » telles que les alcalases, la flavourzyme, etc. Le milieu de culture ainsi hydrolyse est centrifugé, filtré jusqu'à l'obtention d'un hydrolysat de protéines. La solution obtenue constitue l'extrait actif. L'extrait actif peut être encore purifié par fractionnement, notamment par une méthode de type chromatogràphique ou purifié et concentré par un procédé de dialyse. L'une quelconque des formes plus ou moins purifiées de l'extrait est alors solubilisée dans de l'eau ou dans tout mélange contenant de l'eau, puis stérilisée par ultrafiltration. L'extrait végétal obtenu selon l'invention est analysé qualitativement et quantitativement pour sa teneur en composés de nature protéique et peptidique. On entend par composés de nature protéique et peptidique, les fragments de protéines, les peptides et les acides aminés libres présents dans le mélange. Les peptides, acides aminés et fragments de protéines sont dosés selon les techniques classiques, bien connues de l'homme du métier. Ainsi, selon un mode de réalisation avantageux de l'invention, l'extrait végétal actif contient une quantité de composés de nature protéique et peptidique représentant entre 30 et 90 % du poids total de la matière sèche, plus particulièrement cette quantité est comprise entre 60 et 80 % du poids total de la matière sèche, pour les extraits de soja, maïs et tournesol , ou plus particulièrement cette quantité est comprise entre 50 à 80% pour les extraits de lin et de colza.The isolated protein fraction according to the invention is then hydrolyzed under mild conditions to generate polypeptides and soluble peptides. Hydrolysis is defined as a chemical reaction involving the cleavage of a molecule by water, this reaction being possible in a neutral, acidic or basic medium. According to the invention, the hydrolysis is carried out chemically and / or advantageously by proteolytic enzymes. The use of endoproteases and exopeptidases of plant origin (papain, bromelain, ficin) originating from microorganisms may be mentioned. (Aspergillus, Rhizopus, Bacillus, etc.), or by so-called "industrial" enzymes such as alkalases, flavourzyme, etc. The culture medium thus hydrolyzed is centrifuged and filtered until a protein hydrolyzate is obtained. The solution obtained constitutes the active extract. The active extract may be further purified by fractionation, in particular by a chromatographic or purified method and concentrated by a dialysis method. Any of the more or less purified forms of the extract is then solubilized in water or in any mixture containing water, and then sterilized by ultrafiltration. The plant extract obtained according to the invention is analyzed qualitatively and quantitatively for its content of compounds of protein and peptide nature. Protein and peptide compounds are understood to mean protein fragments, peptides and free amino acids present in the mixture. The peptides, amino acids and protein fragments are determined according to conventional techniques, which are well known to those skilled in the art. Thus, according to an advantageous embodiment of the invention, the active plant extract contains a quantity of protein and peptide compounds representing between 30 and 90% of the total weight of the dry matter, more particularly this amount is between 60 and 90% of the total weight of the dry matter. and 80% of the total weight of the dry matter, for extracts of soybean, corn and sunflower, or more particularly that amount is between 50 to 80% for extracts of flax and rapeseed.
L'invention a pour deuxième objet l'utilisation d'un extrait végétal selon l'invention pour la préparation d'une composition pharmaceutique ; l'extrait ou la composition étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de Pépiderme ou du bulbe pileux afin de repigmenter la peau dépigmentée par exemple dans le cas du vitiligo, ou afin de pigmenter les poils ou les cheveux, en particulier dans le traitement et la prévention de la canitie.The subject of the invention is the use of a plant extract according to the invention for the preparation of a pharmaceutical composition; the extract or the composition being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or of the hair bulb in order to repigment the depigmented skin, for example in the case of vitiligo, or in order to pigment hair or hair , especially in the treatment and prevention of canities.
L'invention se rapporte aussi à l'utilisation d'un extrait végétal selon l'invention pour la préparation d'une composition pharmaceutique ; l'extrait ou la composition étant destinés à atténuer les désordres pigmentaires associés à une hypopigmentation de la peau, notamment le vitiligo.The invention also relates to the use of a plant extract according to the invention for the preparation of a pharmaceutical composition; the extract or the composition being intended to attenuate the pigmentary disorders associated with skin hypopigmentation, in particular vitiligo.
L'invention a également pour objet l'utilisation dans une composition cosmétique, ou pour la préparation d'une composition pharmaceutique, d'un extrait végétal selon l'invention ; l'extrait ou la composition étant destinés à favoriser la pigmentation des poils et/ou des cheveux.The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended to promote the pigmentation of the hairs and / or the hair.
L'invention se rapporte encore à l'utilisation dans une composition cosmétique, ou pour la préparation d'une composition pharmaceutique, d'un extrait végétal selon l'invention ; l'extrait ou la composition étant destinés à préparer la peau à une exposition au soleil.The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended to prepare the skin for exposure to the sun.
L'invention se rapporte encore à l'utilisation dans une composition cosmétique, ou pour la préparation d'une composition pharmaceutique, d'un extrait végétal selon l'invention ; l'extrait ou la composition étant destinés, par l'augmentation de la synthèse de mélanine, à protéger la peau des rayonnements du soleil.The invention also relates to the use in a cosmetic composition, or for the preparation of a pharmaceutical composition, of a plant extract according to the invention; the extract or the composition being intended, by increasing melanin synthesis, to protect the skin from sunlight.
Selon une autre forme de l'invention, l'utilisation d'un extrait selon l'invention ou la composition le contenant sont destinés à protéger la peau des stress environnementaux.According to another form of the invention, the use of an extract according to the invention or the composition containing it is intended to protect the skin from environmental stresses.
Selon un mode de réalisation avantageux de l'invention, l'extrait est préalablement solubilisé dans un ou plusieurs solvants cosmétiquement ou pharmaceutiquement acceptables comme l'eau, le glycérol, le propylène glycol, le butylène glycol, les diglycols éthoxylés ou propoxylés, Péthanol, le propanol ou Pisopropanol.According to an advantageous embodiment of the invention, the extract is first solubilized in one or more cosmetically or pharmaceutically acceptable solvents such as water, glycerol, propylene glycol, butylene glycol, ethoxylated or propoxylated diglycols, ethanol, propanol or isopropanol.
Selon encore un autre mode de réalisation avantageux de l'invention, l'extrait est préalablement solubilisé dans un vecteur cosmétique ou pharmaceutique comme les liposomes ou adsorbé sur des polymères organiques poudreux, des supports minéraux comme les talcs et bentonites, et plus généralement solubilisé dans, ou fixé sur, tout vecteur cosmétiquement ou pharmaceutiquement acceptable.According to yet another advantageous embodiment of the invention, the extract is first solubilized in a cosmetic or pharmaceutical vector such as liposomes or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites, and more generally solubilized in or attached to any cosmetically or pharmaceutically acceptable carrier.
Pour donner un ordre de grandeur, dans les compositions cosmétiques ou dermatologiques de la présente invention, les extraits de tournesol et de maïs sont utilisés en une quantité représentant de 0,0001 % à 20 % du poids total de la composition, et préférentiellement en une quantité représentant de 0,05 % à 10 % du poids total de la composition.To give an order of magnitude, in the cosmetic or dermatological compositions of the present invention, the sunflower and corn extracts are used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in one amount representing from 0.05% to 10% of the total weight of the composition.
Dans les mêmes conditions, les extraits de lin, de soja et de colza sont utilisés en une quantité représentant de 0,0001 % à 20 % du poids total de la composition, et préférentiellement en une quantité représentant de 0,05 % à 5 % du poids total de la composition. Ainsi, la teneur en composés de nature protéique et peptidique, dans les compositions de la présente invention, sera alors de 0,00003 % à 18% et préférentiellement de 0,015 % à 9 % du poids total de la composition, pour des extraits végétaux de tournesol et de maïs, contenant entre 30 et 90 % de composés de nature protéique et peptidique .Under the same conditions, the flax, soya and rapeseed extracts are used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing from 0.05% to 5% of the total weight of the composition. Thus, the content of protein and peptide compounds in the compositions of the present invention will then be from 0.00003% to 18% and preferably from 0.015% to 9% of the total weight of the composition, for plant extracts of the present invention. sunflower and corn, containing between 30 and 90% of compounds of protein and peptide nature.
La teneur en composés de nature protéique et peptidique dans les compositions de la présente invention, sera alors de 0,0006 % à 16 % et préférentiellement de 0,03 % à 8 % du poids total de la composition, pour des extraits végétaux de tournesol et de maïs, contenant entre 60 et 80 % de composés de nature protéique et peptidique. La teneur en composés de nature protéique et peptidique, dans les compositions de la présente invention, sera alors de 0,00003 % à 18% et préférentiellement de 0,015 % à 9 % du poids total de la composition, pour des extraits végétaux de lin, de soja et de colza, contenant entre 30 et 90 % de composés de nature protéique et peptidique . La teneur en composés de nature protéique et peptidique dans les compositions de la présente invention, sera alors de 0,0006 % à 16 % et préférentiellement de 0,03 % à 4 % du poids total de la composition, pour des extraits végétaux de soja, contenant entre 60 et 80 % de composés de nature protéique et peptidique.The content of protein and peptide compounds in the compositions of the present invention will then be from 0.0006% to 16% and preferably from 0.03% to 8% of the total weight of the composition, for plant extracts of sunflower and corn, containing between 60 and 80% of compounds of protein and peptide nature. The content of protein and peptide compounds in the compositions of the present invention will then be from 0.00003% to 18% and preferably from 0.015% to 9% of the total weight of the composition, for plant extracts of flax, soybean and rapeseed, containing between 30 and 90% of compounds of protein and peptide nature. The content of protein and peptide compounds in the compositions of the present invention will then be from 0.0006% to 16% and preferably from 0.03% to 4% of the total weight of the composition, for plant extracts of soya , containing between 60 and 80% of compounds of protein and peptide nature.
La teneur en composés de nature protéique et peptidique dans les compositions de la présente invention, sera alors de 0,0005 % à 16 % et préférentiellement de 0,025 % à 4 % du poids total de la composition, pour des extraits végétaux de lin et de colza, contenant entre 50 et 80 % de composés de nature protéique et peptidique.The content of protein and peptide compounds in the compositions of the present invention will then be from 0.0005% to 16% and preferably from 0.025% to 4% of the total weight of the composition, for plant extracts of flax and rapeseed, containing between 50 and 80% of compounds of protein and peptide nature.
Outre l'intérêt majeur de la pigmentation de la peau dans la protection solaire, un composé pro-pigmentant selon l'invention offre des débouchés dans les produits auto-bronzants et les produits favorisant le bronzage. Pour cela, l'invention se rapporte à l'utilisation d'extrait pour la fabrication de produits pour le bronzage et/ou le brunissage artificiels de la peau.In addition to the major advantage of the pigmentation of the skin in the sun protection, a pro-pigmenting compound according to the invention offers outlets in self-tanning products and products promoting tanning. For this, the invention relates to the use of extract for the manufacture of products for artificial tanning and / or browning of the skin.
L'invention se rapporte aussi à l'utilisation d'extrait pour améliorer l'intensité et/ou l'homogénéité et/ou la persistance de la coloration produite sur la peau et/ou les cheveux.The invention also relates to the use of extract to improve the intensity and / or the homogeneity and / or the persistence of the coloration produced on the skin and / or the hair.
Préférentiellement, les compositions selon la présente invention se présenteront sous une forme adaptée à l'administration par voie topique cutanée, et couvrent toutes les formes cosmétiques ou dermatologiques. Ces compositions doivent donc contenir un milieu cosmétiquement et/ou dermatologiquement acceptable, c'est-à-dire compatible avec la peau, les poils ou les cheveux. Ces compositions pourront notamment être sous forme de crèmes, émulsions H/E, E/H ou émulsions multiples, solutions, suspensions, gels, laits, lotions, sticks ou encore poudres, adaptés à une application sur la peau, les lèvres et/ou les cheveux.Preferably, the compositions according to the present invention will be in a form suitable for topical administration to the skin, and cover all cosmetic or dermatological forms. These compositions must therefore contain a medium cosmetically and / or dermatologically acceptable, that is to say compatible with the skin, hair or hair. These compositions may especially be in the form of creams, O / W emulsions, W / O emulsions or multiple emulsions, solutions, suspensions, gels, milks, lotions, sticks or even powders, suitable for application to the skin, lips and / or the hair.
Ces compositions comprennent les excipients nécessaires à leur formulation, tels que solvants, épaississants, diluants, tensioactifs, anti-oxydants, colorants, conservateurs, parfums.These compositions comprise the excipients necessary for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes.
Bien entendu, l'homme de métier veillera à choisir les éventuels composés complémentaires, actifs ou non-actifs, et/ou leur quantité, de telle sorte que les propriétés avantageuses du mélange ne soient pas, ou sensiblement pas, altérées par l'adjonction envisagée.Of course, those skilled in the art will take care to choose any additional compounds, active or non-active, and / or their quantity, so that the advantageous properties of the mixture are not, or not substantially, impaired by the addition considered.
La composition utilisable selon l'invention peut en particulier consister en une composition pour soins capillaires, et notamment un shampooing, un après-shampooing, une lotion de mise en plis, une lotion traitante, une crème ou un gel coiffant, une lotion restructurante pour les cheveux, un masque, etc. La composition cosmétique selon l'invention peut être utilisée notamment dans les traitements mettant en oeuvre une application qui est suivie ou non suivie d'un rinçage, ou encore sous forme de shampooing. Elle peut également se présenter sous forme de teinture ou de mascara à appliquer au pinceau ou au peigne, en particulier sur les cils, les sourcils ou les cheveux.The composition that may be used according to the invention may in particular consist of a composition for hair care, and in particular a shampoo, a conditioner, a setting lotion, a treatment lotion, a cream or a styling gel, a restructuring lotion for hair, a mask, etc. The cosmetic composition according to the invention can be used in particular in treatments using an application that is followed or not followed by rinsing, or in the form of shampoo. It can also be in the form of dye or mascara to be applied with a brush or a comb, in particular on eyelashes, eyebrows or hair.
Il est bien entendu évident que les extraits végétaux, principalement constitués de composés de nature protéique et peptidique, selon l'invention pourront être utilisés en tant que seul agent pro-pigmentant ou bien en association avec au moins un autre agent actif, dans une composition cosmétique ou pour la préparation d'une composition dermatologique et/ou pharmaceutique.It is, of course, obvious that the plant extracts, mainly consisting of protein and peptide compounds, according to the invention may be used as the only pro-pigmenting agent or in combination with at least one other active agent, in a composition cosmetic or for the preparation of a dermatological and / or pharmaceutical composition.
Avantageusement, les compositions utilisables selon l'invention contiennent en outre au moins un autre agent favorisant la pigmentation de la peau, des cheveux et/ou des poils. De tels composés sont notamment des substrats de la tyrosinase, tels que la tyrosine ou la L-DOPA, des prostaglandines, ou des composés activateurs de la voie de l'AMPc tels que des dérivés de pro-opiomélanocortines, l'adénosine, ou la forkoline ou ses dérivés. On peut également citer des extraits de végétaux tels que le bigaradier (citrus aurantium) ou de chrysanthème (Chrysanthemum morifoliurrî), décrits notamment dans les brevets FR 2845285 et EP1014934.Advantageously, the compositions that can be used according to the invention also contain at least one other agent that promotes the pigmentation of the skin, hair and / or hair. Such compounds are in particular tyrosinase substrates, such as tyrosine or L-DOPA, prostaglandins, or activating compounds of the cAMP pathway such as pro-opiomelanocortin derivatives, adenosine, or forkoline or its derivatives. Plant extracts such as bitter orange (citrus aurantium) or chrysanthemum (Chrysanthemum morifoliurri), described in particular in patents FR 2845285 and EP1014934.
De tels composés se trouvent également dans la famille des colorants exogènes des couches superficielles de l'épiderme, tels que la dihydroxyacétone (DHA), Pérythrulose, les extraits de feuilles de henné, décrits notamment dans les brevets EP 0742002, FR2779958.Such compounds are also found in the family of exogenous dyes of the superficial layers of the epidermis, such as dihydroxyacetone (DHA), perythrulose, extracts of henna leaves, described in particular in patents EP 0742002 and FR2779958.
D'autres avantages et caractéristiques de l'invention apparaîtront mieux à la lecture des exemples donnés à titre illustratif et non limitatif.Other advantages and characteristics of the invention will appear better on reading the examples given by way of illustration and not limitation.
Exemple 1 : Préparation d'un extrait de lin (Linum usitatissimum L.) Dans une première étape, 1 kg de graines de lin sont délipidées par l'action d'un solvant organique : Phexane. Après séchage du produit, on obtient un résidu enrichi en protéines (le tourteau). Le résidu est alors dilué dans une solution aqueuse légèrement basique puis hydrolyse par ajout d'une enzyme : la papaïne, à 60°C pendant 4 heures sous agitation. Ce mélange est ensuite chauffé à 80 °C, centrifugé et filtré jusqu'à l'obtention d'une solution limpide. Il est ensuite concentré sous vide, puis filtré de nouveau sur des filtres-plaques puis sur cartouche stérilisante. On obtient alors un hydrolysat titrant à 30 g/1 de composés de nature protéique. L'extrait hydrolyse contient une quantité de composés de nature protéique représentant environ 65 % du poids total de la matière sèche. Cet hydrolysat est remis en solution dans du dipropylène glycol.Example 1 Preparation of a Linseed Extract (Linum usitatissimum L.) In a first step, 1 kg of flaxseed is delipidated by the action of an organic solvent: hexane. After drying the product, a residue enriched in proteins (the cake) is obtained. The residue is then diluted in a slightly basic aqueous solution and then hydrolyzed by adding an enzyme: papain, at 60 ° C. for 4 hours with stirring. This mixture is then heated to 80 ° C, centrifuged and filtered until a clear solution is obtained. It is then concentrated under vacuum, then filtered again on filter plates and sterilizing cartridge. A hydrolyzate titrating at 30 g / l of compounds of protein nature is then obtained. The hydrolyzed extract contains a quantity of protein compounds representing approximately 65% of the total weight of the dry matter. This hydrolyzate is redissolved in dipropylene glycol.
Exemple 2 : Préparation d'extraits déphénolisés à partir de maïs (Zea mays) 100 grammes de tourteau de maïs sont dans un premier temps déphénolisés jusqu'à obtention d'une teneur en phénols inférieure à 0,005 %.Example 2 Preparation of Dephenolated Extracts from Corn (Zea Mays) 100 grams of maize cake are initially dephenolized to a phenol content of less than 0.005%.
Puis les protéines sont précipitées afin d'augmenter leur concentration.Then the proteins are precipitated in order to increase their concentration.
Ensuite, le produit est hydrolyse enzymatiquement en milieu aqueux par un cocktail enzymatique (protéase, lipase et cellulase) à différents pH et différentes températures pendant 4 à 8 heures puis filtré. Des charbons actifs sont ensuite utilisés pour obtenir un produit qui satisfasse les critères cosmétiques en terme d'odeur et de couleur. Le produit est enfin stérilisé par fîltration stérilisante et chauffage.Then, the product is enzymatically hydrolyzed in aqueous medium by an enzymatic cocktail (protease, lipase and cellulase) at different pH and temperatures for 4 to 8 hours and then filtered. Activated carbons are then used to obtain a product that satisfies the cosmetic criteria in terms of odor and color. The product is finally sterilized by sterilizing filtration and heating.
L'hydrolysat ainsi obtenu contient plus de 80 % de protéines et moins de 0,005 % de phénols. Exemple 3 : Préparation d'extraits déphénolisés à partir de tournesol (Helianthus annuus) 100 grammes de tourteau de tournesol sont dans un premier temps déphénolisés jusqu'à obtention d'une teneur en phénols inférieure à 0,005 %. Puis les protéines sont précipitées afin d'augmenter leur concentration. Ensuite, le produit est hydrolyse enzymatiquement en milieu aqueux par un cocktail enzymatique (protéase, lipase et cellulase) à différents pH et différentes températures pendant 4 à 8 heures puis filtré. Des charbons actifs sont ensuite utilisés pour obtenir un produit qui satisfasse les critères cosmétiques en terme d'odeur et de couleur. Le produit est enfin stérilisé par filtration stérilisante et chauffage. L'hydrolysat ainsi obtenu contient plus de 80 % de protéines et moins de 0,005 % de phénols.The hydrolyzate thus obtained contains more than 80% of proteins and less than 0.005% of phenols. EXAMPLE 3 Preparation of Dephenolated Extracts from Sunflower (Helianthus annuus) 100 grams of sunflower cake are initially dephenolized until a phenol content of less than 0.005% is obtained. Then the proteins are precipitated in order to increase their concentration. Then, the product is enzymatically hydrolyzed in aqueous medium by an enzymatic cocktail (protease, lipase and cellulase) at different pH and temperatures for 4 to 8 hours and then filtered. Activated carbons are then used to obtain a product that satisfies the cosmetic criteria in terms of odor and color. The product is finally sterilized by sterilizing filtration and heating. The hydrolyzate thus obtained contains more than 80% of proteins and less than 0.005% of phenols.
Exemple 4 : Préparation d'extraits à partir de tourteau de soja {Glycine max L.)EXAMPLE 4 Preparation of Extracts From Soybean Meal (Glycine max L.)
Les tourteaux sont les résidus solides obtenus après extraction de l'huile des graines de soja. Ils représentent de 50 à 75 % de la masse des graines. Les tourteaux sont mis en suspension dans une solution aqueuse alcaline (dilution au 1/10). Ce mélange est maintenu sous agitation pendant un temps suffisamment long pour permettre la solubilisation des fractions solubles. La température d'extraction est variable (comprise entre 4 et 80°C) ; préférentiellement l'opération sera réalisée à froid. Après cette phase d'extraction le milieu est clarifié par centrifugation puis filtré sur filtre à plaque. Ce filtrat qui contient les fractions solubles du soja est ensuite soumis à une précipitation des protéines en faisant varier la force ionique en milieu neutre ou acide, ce qui permet d'éliminer les composants glucidiques solubles, les lipides et les acides nucléiques. Le surnageant est éliminé et le précipité est ensuite lavé à l'aide d'un solvant tel que, par exemple, l'éthanol ou le méthanol puis le solvant est évaporé par séchage sous vide.The cakes are the solid residues obtained after extraction of the oil from the soya beans. They represent 50 to 75% of the seed mass. The cakes are suspended in an aqueous alkaline solution (1/10 dilution). This mixture is stirred for a time long enough to allow the solubilization of the soluble fractions. The extraction temperature is variable (between 4 and 80 ° C); preferably the operation will be performed cold. After this extraction phase, the medium is clarified by centrifugation and then filtered through a plate filter. This filtrate, which contains the soluble fractions of soybeans, is then precipitated by varying the ionic strength in a neutral or acidic medium to remove soluble carbohydrate components, lipids, and nucleic acids. The supernatant is removed and the precipitate is then washed with a solvent such as, for example, ethanol or methanol and the solvent is evaporated by drying in vacuo.
A ce stade, on obtient environ 50 grammes de poudre de couleur jaune clair d'extrait protéique brut contenant : Protéines : 75 % Glucides : 20 % - Lipides : 5 %At this stage, about 50 grams of light yellow powder of crude protein extract are obtained containing: Proteins: 75% Carbohydrates: 20% - Lipids: 5%
Le précipité riche en protéines est remis en solution dans l'eau ou un autre solvant. L'extraction peut également être réalisée en milieu neutre ou acide. L'extrait protéique brut est alors soumis à une série d'hydrolyses ménagées et sélectives consistant en des hydrolyses chimiques et enzymatiques. A titre d'exemple on peut citer l'hydrolyse en présence d'exopeptidases (papaïne, ficine).The protein-rich precipitate is dissolved in water or another solvent. The extraction can also be carried out in neutral or acidic medium. The crude protein extract is then subjected to a series of gentle and selective hydrolyses consisting of chemical and enzymatic hydrolyses. By way of example, mention may be made of hydrolysis in the presence of exopeptidases (papain, ficin).
L'hydrolysat est ensuite purifié puis dialyse sur membrane pour éliminer les composés de haut poids moléculaire.The hydrolyzate is then purified and membrane-dialyzed to remove the high molecular weight compounds.
On obtient alors un hydrolysat de couleur claire titrant de 15 à 30 g/1 d'extrait sec et contenant une quantité de composés de nature protéique et peptidique représentant environ 70 % du poids total de la matière sèche. L'hydrolysat est alors dilué de telle sorte que la concentration en protéines déterminée par la méthode de Lowry, soit comprise entre 0,5 et 2 g/1.A light-colored hydrolyzate titrant of 15 to 30 g / l of dry extract and containing a quantity of compounds of protein and peptide nature representing approximately 70% of the total weight of the dry matter is then obtained. The hydrolyzate is then diluted so that the protein concentration determined by the Lowry method is between 0.5 and 2 g / l.
Exemple 5 : Préparation d'un extrait de colza (Brassica napus).Example 5: Preparation of an extract of rapeseed (Brassica napus).
Dans une première étape, 1 kg des graines de colza sont délipidées par l'action d'un solvant organique : Phexane. Après séchage du produit, on obtient un résidu enrichi en protéines (le tourteau). Le tourteau est alors dilué dans une solution aqueuse légèrement basique puis hydrolyse par ajout d'une enzyme : la papaïne, à 600C pendant 4 heures sous agitation. Ce mélange est ensuite chauffé à 80 0C, centrifugé et filtré jusqu'à l'obtention d'une solution limpide. Il est ensuite concentré sous vide, puis filtré de nouveau sur des filtres-plaques puis sur cartouche stérilisante. On obtient alors un hydrolysat de couleur brune titrant à 30 g/1 de composés de nature protéique. L'extrait hydrolyse contient une quantité de composés de nature protéique représentant environ 65 % du poids total de la matière sèche. Cet hydrolysat est remis en solution dans du dipropylène glycol.In a first step, 1 kg of the rapeseeds are delipidated by the action of an organic solvent: hexane. After drying the product, a residue enriched in proteins (the cake) is obtained. The cake is then diluted in a slightly basic aqueous solution and then hydrolyzed by adding an enzyme: papain, at 60 ° C. for 4 hours with stirring. This mixture is then heated to 80 ° C., centrifuged and filtered until a clear solution is obtained. It is then concentrated under vacuum, then filtered again on filter plates and sterilizing cartridge. A brown-colored hydrolyzate titrating at 30 g / l of protein-like compounds is then obtained. The hydrolyzed extract contains a quantity of protein compounds representing approximately 65% of the total weight of the dry matter. This hydrolyzate is redissolved in dipropylene glycol.
Exemple 6 : Etude ex vivo de l'effet des extraits selon les exemples 1 à 5 sur la synthèse de mélanineEXAMPLE 6 Ex Vivo Study of the Effect of the Extracts According to Examples 1 to 5 on the Synthesis of Melanin
Le but de cette étude ex vivo est de mettre en évidence l'augmentation de la mélanisation apportée par les extraits selon les exemples 1 à 5. Protocole : Des biopsies de peau humaine de 6 mm de diamètre sont maintenues en culture ex vivo en présence d'un milieu spécifique (DMEM lg/L, HAMF 12, SVF et antibiotiques) sur des inserts déposés dans des plaques 6 puits. Les biopsies sont cultivées pendant 48 heures et reçoivent ou non 2 applications par jour d'extrait selon les exemples 1 à 5, à une concentration de 1 %. Les biopsies sont ensuite fixées dans le formol 9 %, NaCl (150 mM) pendant 10 heures puis incluses dans la paraffine. Des coupes de peau de 3 μm d'épaisseur sont ensuite réalisées et la mélanine est colorée spécifiquement par la technique de Fontana-Masson.The purpose of this ex vivo study is to demonstrate the increase in melanization provided by the extracts according to Examples 1 to 5. Protocol: Human skin biopsies 6 mm in diameter are maintained in culture ex vivo in the presence of a specific medium (DMEM lg / L, HAMF 12, FCS and antibiotics) on inserts deposited in 6-well plates. The biopsies are cultured for 48 hours and receive or not 2 applications per day of extract according to Examples 1 to 5, at a concentration of 1%. The biopsies are then fixed in 9% formalin, NaCl (150 mM) for 10 hours and then included in the paraffin. 3 μm thick skin sections are then made and the melanin is colored specifically by the Fontana-Masson technique.
Résultats : Les coupes de peau n'ayant pas reçu l'application d'extrait selon les exemples 1 à 5 présentent une coloration de faible intensité. Au contraire, les coupes de peau ayant reçu les applications d'extrait selon les exemples 1 à 5 présentent une coloration d'intensité nettement augmentée. De plus, la mélanine est située dans la couche basale, mais aussi transportée dans les couches supra-basales.Results: The skin sections that did not receive the application of extract according to Examples 1 to 5 have a low intensity coloration. On the contrary, the skin sections having received the extract applications according to Examples 1 to 5 show a markedly increased color intensity. In addition, melanin is located in the basal layer, but also transported in the supra-basal layers.
Conclusions : 48 heures de traitement ont permis d'induire une forte augmentation de la synthèse de mélanine par les mélanocytes et de stimuler l'ensemble du processus de répartition de la mélanine dans Pépiderme.Conclusions: 48 hours of treatment allowed to induce a strong increase of melanin synthesis by melanocytes and to stimulate the whole process of distribution of melanin in the epidermis.
Exemple 7 : Préparation de compositionsExample 7 Preparation of compositions
Ces compositions ont été obtenues par simple mélange des différents composants. Les quantités indiquées sont données en pourcentage de poids.These compositions were obtained by simple mixing of the various components. The quantities indicated are given as a percentage of weight.
1-Emulsion huile-dans-eau1-oil-in-water emulsion
Phase huileuse :Oily phase:
B Montanov 68 (Cetearyl Alcohol and Cetearyl Glucoside) 5.00 %B Montanov 68 (Cetearyl Alcohol and Cetearyl Glucoside) 5.00%
" Huile de Jojoba 5.00 % u Huile de Vaseline 5.00 %"Jojoba oil 5.00% u Vaseline oil 5.00%
" Isopropyl Palmitate 7.00 %"Isopropyl Palmitate 7.00%
Phase aqueuse :Aqueous phase :
" Glycérine 5.00 % m Allantoïne 0.10 % m Extrait selon l'exemple 1 1.00 %"Glycerin 5.00% m Allantoin 0.10% m Extract according to Example 1 1.00%
" Sepigel 305 (Polyacrylamide and C13-14 Isoparaffm and Laureth-7) 0.30 %Sepigel 305 (Polyacrylamide and C13-14 Isoparaffm and Laureth-7) 0.30%
" Conservateur 0.50 %"Conservative 0.50%
" Parfum 0.50 % m Eau qsp 100 %"Perfume 0.50% m Water qs 100%
2- Gel2- Gel
" Carbopol Ultrez lO (sol. à 2% ) 25.00 %"Carbopol Ultrez 10 (2% solids) 25.00%
B Triéthanolamine 0.50 %B Triethanolamine 0.50%
" Extrait selon l'exemple 2 5.00 % m Conservateur 0.20 %"Extract according to example 2 5.00% m Conservative 0.20%
" EDTA (séquestrant) 0.10 % m Parfum 0.50 % m Eau qsp 100 %"EDTA (sequestering) 0.10% m Perfume 0.50% m Water qs 100%
3- Lotion m Mono Propylène Glycol 1.00 % m Allantoïne 0.30 %3- Lotion m Mono Propylene Glycol 1.00% m Allantoin 0.30%
M Glycérine 1.00 % - Cetiol HE (PEG-7 Glyceryl Cocoate) 1.00 %M Glycerin 1.00% - Cetiol HE (PEG-7 Glyceryl Cocoate) 1.00%
« Extrait selon l'exemple 3 0.5 % m Conservateur 0.20 %"Extract according to example 3 0.5% m Preservative 0.20%
M Parfum 0.50 %M Perfume 0.50%
" Eau qsp 100 %"Water qs 100%
4-Emulsion autobronzante huile-dans-eau Phase huileuse : β Mélange mono/distéarate de glycérol 2.00 % B Alcool stéarylique 1.00 % m Acide stéarique d'huile de palme 1.50 %4-Oil-in-water self-tanning emulsion Oily phase: β Glycerol mono / distearate mixture 2.00% B Stearyl alcohol 1.00% m Palm oil stearic acid 1.50%
• PoIy diméthylsiloxane 2.00 %• Dimethylsiloxane poIy 2.00%
• Mélange poly diméthylsiloxane alpha-omega 3.00 % dihydroxyle/cyclopentadiméthylsiloxane (14,7/85,3)• Poly dimethylsiloxane mixture alpha-omega 3.00% dihydroxyl / cyclopentadimethylsiloxane (14.7 / 85.3)
• Lauroyl sarconisate d'isopropyle 15.00 % Phase aqueuse :• Lauroyl isopropyl sarconisate 15.00% Aqueous phase:
" Dihydroxyacétone 5.00 %"Dihydroxyacetone 5.00%
B Glycérine 5.00 %B Glycerin 5.00%
• Phosphate d'alcool hexadecylique, sel de potassium 1.00 %• Hexadecyl alcohol phosphate, potassium salt 1.00%
• Acide polyacrylique 0.30 % m Triéthanolamine qsp pH : 7• Polyacrylic acid 0.30% m Triethanolamine qsp pH: 7
M Acétate de dl-alpha-tocopheryle 0.50 % M Extrait selon l'exemple 4 1.00 % B Eau déminéralisée qsp 100.00 % 5-Emulsion autobronzante eau-dans-huileM Dl-alpha-tocopheryl acetate 0.50% M Extract according to Example 4 1.00% B Demineralized water qs 100.00% 5-Self-tanning water-in-oil emulsion
Phase huileuse :Oily phase:
B Polydiméthyl/methylcetyl methylsiloxane oxyéthyléné 2.00 % B Phényl triméthylsiloxy trisiloxane 3.00 %B Polydimethyl / methylcetyl methylsiloxane oxyethylenated 2.00% B Phenyl trimethylsiloxy trisiloxane 3.00%
B Lauroyl sarcosinate d'isopropyle 10.00 %B Lauroyl isopropyl sarcosinate 10.00%
B Isohexadecane 5.00 %B Isohexadecane 5.00%
B Cyclopenta dimethylsiloxane 3.00 %B Cyclopenta dimethylsiloxane 3.00%
Phase aqueuse : M Dihydroxyacétone 2.00 %Water phase: M Dihydroxyacetone 2.00%
B Glycérine 5.00 %B Glycerin 5.00%
• Sulfate de magnésium 0.70 %• Magnesium sulphate 0.70%
B Extrait selon l'exemple 5 1.00 %B Extract according to example 5 1.00%
B Eau déminéralisée qsp 100.00 %B Demineralized water qs 100.00%
6- Lotion autobronzante " Dihydroxyacétone 5.00 %6- Self tanning lotion "Dihydroxyacetone 5.00%
« Extrait lyophilisé d' Εclipta prostata 0.40 % m Ethanol à 96% 13.00 % u Ethoxydiglycol 7.00 %"Lyophilized extract of Εclipta prostata 0.40% m 96% ethanol 13.00% u Ethoxydiglycol 7.00%
" PPG-l-PEG-9-Lauryl Glycol Ether 5.00 %"PPG-1-PEG-9-Lauryl Glycol Ether 5.00%
B Parabens 0.25 %B Parabens 0.25%
B Extrait selon l'exemple 2 1.00 %B Extract according to example 2 1.00%
B Eau déminéralisée qsp 100.00 %B Demineralized water qs 100.00%
7- Emulsion huile-dans-eau autobronzante B Eau 66.80 %7- Self-tanning oil-in-water emulsion B Water 66.80%
B Ethoxydiglycol 5.00 %B Ethoxydiglycol 5.00%
B Xanthan gum 0.30 % B Cetyl hydroxyethylcellulose 0.30 %B Xanthan gum 0.30% B Cetyl hydroxyethylcellulose 0.30%
B Cetyl alcohol 2.00 %B Cetyl alcohol 2.00%
B Stearyl alcohol 2.00 %B Stearyl alcohol 2.00%
B Octyl palmitate 2.00 %B Octyl palmitate 2.00%
B Glucam P-20 distearate 2.00 % • Steareth-20 1.00 % " Dimethicone 1.00 % " Polysorbate 60 1.00 % β Dimethicone Copolyol/IPDI Copolymer 1.00 % β Glycerylstearate and PEG-100 stéarate 0.25 % " Actiplex 335 Lipo OP 0.1 % " Tocopheryl acétate 0.1 % " Eau 10.975 %B Glucam P-20 distearate 2.00% • Steareth-20 1.00% "Dimethicone 1.00%" Polysorbate 60 1.00% β Dimethicone Copolyol / IPDI Copolymer 1.00% β Glycerylstearate and PEG-100 Stearate 0.25% "Actiplex 335 Lipo OP 0.1%" Tocopheryl Acetate 0.1% "Water 10.975%
• Dihydroxyacetone 2.00 % M Nipaguard MPA 0.60 %• Dihydroxyacetone 2.00% M Nipaguard MPA 0.60%
" Panthenol, 50% aqueux 0.40 %"Panthenol, 50% aqueous 0.40%
" Sorbic acid 0.05 % β Sodium metabisulfite 0.025 %"Sorbic acid 0.05% β Sodium metabisulphite 0.025%
" Extrait selon l'exemple 3 1.00 % "Extract according to example 3 1.00%

Claims

REVENDICATIONS
1. Utilisation d'un extrait végétal obtenu à partir de plantes choisies parmi les genres Linum, Zea, Helianthus ou les espèces Glycine max et Brassica napus, principalement constitué de composés de nature protéique et peptidique, seul ou en association avec au moins un autre agent actif, dans une composition cosmétique, l'extrait ou la composition le contenant étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épiderme ou du bulbe pileux et à améliorer la pigmentation naturelle de la peau.1. Use of a plant extract obtained from plants selected from the genera Linum, Zea, Helianthus or the species Glycine max and Brassica napus, mainly consisting of protein and peptide compounds, alone or in combination with at least one other active agent in a cosmetic composition, the extract or the composition containing it being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb and to improve the natural pigmentation of the skin.
2. Utilisation d'un extrait végétal obtenu à partir de plantes choisies parmi les genres Linum, Zea, Helianthus ou les espèces Glycine max et Brassica napus, principalement constitué de composés de nature protéique et peptidique, seul ou en association avec au moins un autre agent actif, pour la préparation d'une composition dermatologique et/ou pharmaceutique, l'extrait ou la composition le contenant étant destinés à favoriser l'augmentation de la synthèse de mélanine dans les mélanocytes de l'épiderme ou du bulbe pileux et à atténuer les désordres pigmentaires associés à une hypopigmentation de la peau.2. Use of a plant extract obtained from plants selected from the genera Linum, Zea, Helianthus or the species Glycine max and Brassica napus, mainly consisting of protein and peptide compounds, alone or in combination with at least one other active agent, for the preparation of a dermatological and / or pharmaceutical composition, the extract or the composition containing it being intended to promote the increase of melanin synthesis in the melanocytes of the epidermis or hair bulb and to attenuate pigmentary disorders associated with hypopigmentation of the skin.
3. Utilisation selon les revendications 1 ou 2, caractérisée en ce que les plantes appartenant au genre Linum sont de l'espèce Linum usitatissimum L. (lin).3. Use according to claims 1 or 2, characterized in that the plants belonging to the genus Linum are of the species Linum usitatissimum L. (flax).
4. Utilisation selon les revendications 1 ou 2, caractérisée en ce que les plantes appartenant au genre Zea sont de l'espèce Zea mays L. (maïs).4. Use according to claims 1 or 2, characterized in that the plants belonging to the genus Zea are of the species Zea mays L. (corn).
5. Utilisation selon les revendications 1 ou 2, caractérisée en ce que les plantes appartenant au genre Helianthus sont de l'espèce Helianthus annuus L. (tournesol).5. Use according to claims 1 or 2, characterized in that the plants belonging to the genus Helianthus are of the species Helianthus annuus L. (sunflower).
6. Utilisation selon l'une des revendications précédentes, caractérisée en ce que l'extrait végétal contient une quantité de composés de nature protéique et peptidique, représentant entre 30 % et 90 % du poids total de la matière sèche et plus particulièrement entre 60 % et 80 % du poids total de la matière sèche. 6. Use according to one of the preceding claims, characterized in that the plant extract contains a quantity of compounds of protein and peptide nature, representing between 30% and 90% of the total weight of the dry matter and more particularly between 60% and 80% of the total weight of the dry matter.
7. Utilisation selon l'une quelconque des revendications précédentes, caractérisée en ce que l'extrait est utilisé en une quantité représentant de 0,0001 % à 20 % du poids total de la composition, et préférentiellement en une quantité représentant de 0,05 % à 10 % pour les extraits de maïs et de tournesol, ou préférentiellement en une quantité représentant de 0,05 % à 5 % du poids total de la composition, pour les extraits de lin, de soja et de colza.7. Use according to any one of the preceding claims, characterized in that the extract is used in an amount representing from 0.0001% to 20% of the total weight of the composition, and preferably in an amount representing 0.05 % to 10% for the extracts of corn and sunflower, or preferably in an amount representing from 0.05% to 5% of the total weight of the composition, for extracts of flax, soybean and rapeseed.
8. Utilisation selon l'une quelconque des revendications précédentes, caractérisée en ce que la composition se présente sous une forme adaptée à l'application par voie topique comprenant un milieu cosmétiquement ou dermatologiquement acceptable.8. Use according to any one of the preceding claims, characterized in that the composition is in a form suitable for topical application comprising a cosmetically or dermatologically acceptable medium.
9. Utilisation selon l'une quelconque des revendications précédentes, caractérisée en ce que la composition contient en outre au moins un colorant exogène des couches superficielles de l'épidémie et/ou au moins un agent actif pro-pigmentant différent dudit extrait végétal selon les revendications 1 à 6, choisi parmi les substrats de la tyrosinase ou les prostaglandines ou les composés activateurs de la voie de l'AMPc ou les extraits végétaux pigmentants.9. Use according to any one of the preceding claims, characterized in that the composition also contains at least one exogenous dye epidermis surface layers and / or at least one pro-pigmenting active agent different from said plant extract according to the Claims 1 to 6, selected from tyrosinase substrates or prostaglandins or activating compounds of the cAMP pathway or plant extracts pigment.
10. Utilisation selon l'une quelconque des revendications précédentes caractérisée en ce que l'extrait ou la composition le contenant sont destinés, par l'augmentation de la synthèse de mélanine, à préparer la peau à une exposition au soleil ou à protéger la peau des rayonnements du soleil.10. Use according to any one of the preceding claims, characterized in that the extract or the composition containing it is intended, by increasing the synthesis of melanin, to prepare the skin for exposure to the sun or to protect the skin. radiation from the sun.
11. Utilisation selon l'une quelconque des revendications précédentes caractérisée en ce que l'extrait ou la composition le contenant sont destinés à protéger la peau des agressions extérieures, en particulier des rayons UV.11. Use according to any one of the preceding claims characterized in that the extract or composition containing it is intended to protect the skin from external aggressions, in particular UV rays.
12. Utilisation selon l'une quelconque des revendications précédentes caractérisée en ce que l'extrait ou la composition le contenant sont destinés au bronzage et/ou au brunissage artificiels de la peau. 12. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended for artificial tanning and / or browning of the skin.
13. Utilisation selon l'une quelconque des revendications précédentes caractérisée en ce que l'extrait ou la composition le contenant sont destinés à améliorer l'intensité et/ou l'homogénéité et/ou la persistance de la pigmentation de la peau et/ou des cheveux.13. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended to improve the intensity and / or homogeneity and / or the persistence of the pigmentation of the skin and / or hair.
14. Utilisation selon l'une quelconque des revendications précédentes caractérisée en ce que l'extrait ou la composition le contenant sont destinés à prévenir ou à traiter le blanchiment des poils et/ou des cheveux. 14. Use according to any one of the preceding claims characterized in that the extract or the composition containing it are intended to prevent or treat the whitening of hair and / or hair.
PCT/FR2007/001334 2006-08-03 2007-08-02 Use of a plant extract as active agent for increasing melanin synthesis in melanocytes WO2008015342A2 (en)

Applications Claiming Priority (10)

Application Number Priority Date Filing Date Title
FR0607101 2006-08-03
FR0607093 2006-08-03
FR0607096A FR2904541B1 (en) 2006-08-03 2006-08-03 USE OF SOYBEAN EXTRACT AS ACTIVE AGENT FOR INCREASING THE SYNTHESIS OF MELANIN IN MELANOCYTES
FR0607095A FR2904540B1 (en) 2006-08-03 2006-08-03 USE OF SUNFLOWER EXTRACT AS ACTIVE AGENT FOR INCREASING THE SYNTHESIS OF MELANIN IN MELANOCYTES
FR0607106 2006-08-03
FR0607095 2006-08-03
FR0607093A FR2904555B1 (en) 2006-08-03 2006-08-03 USE OF A COLZA EXTRACT AS AN ACTIVE AGENT TO INCREASE THE SYNTHESIS OF MELANIN IN MELANOCYTES
FR0607101A FR2904543B1 (en) 2006-08-03 2006-08-03 USE OF MAIZE EXTRACT AS AN ACTIVE AGENT TO INCREASE MELANIN SYNTHESIS IN MELANOCYTES
FR0607106A FR2904547B1 (en) 2006-08-03 2006-08-03 USE OF FLAX EXTRACT AS ACTIVE AGENT FOR INCREASING THE SYNTHESIS OF MELANIN IN MELANOCYTES
FR0607096 2006-08-03

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WO2010029115A1 (en) * 2008-09-10 2010-03-18 L V M H Recherche Methods useful in studying or modulating skin or hair pigmentation, plant extracts for use in compositions and cosmetic care method
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US8257753B2 (en) 2007-07-20 2012-09-04 Isp Investments Inc. Use of an active principle originating from flax in a composition intended to reinforce the barrier function of the skin and to protect the skin and the appendages against external aggressions
WO2014029948A2 (en) * 2012-08-24 2014-02-27 Isp Investments Inc. Flax extract and cosmetic composition comprising said extract for increasing the level of intracellular coenzyme q10
ITBO20130279A1 (en) * 2013-05-31 2014-12-01 Phenbiox S R L COMPOSITION FOR BODY CARE
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US8257753B2 (en) 2007-07-20 2012-09-04 Isp Investments Inc. Use of an active principle originating from flax in a composition intended to reinforce the barrier function of the skin and to protect the skin and the appendages against external aggressions
US8349803B2 (en) 2007-07-20 2013-01-08 Isp Investments Inc. Use of an active principle from flax for use in a composition for activating cytochrome c
FR2918893A1 (en) * 2007-07-20 2009-01-23 Vincience Sa Use of active ingredient of natural peptide, obtained from hydrolysis of flax protein (Linum), in cosmetic composition or for preparing pharmaceutical composition, e.g. to activate cytochrome c and stimulate mitochondria
WO2009043991A3 (en) * 2007-07-20 2009-11-26 Societe D'extraction Des Principes Actifs S.A. (Vincience) Use of an active principle from flax for use in a composition for activating cytochrome c
WO2009043991A2 (en) 2007-07-20 2009-04-09 Societe D'extraction Des Principes Actifs S.A. (Vincience) Use of an active principle from flax for use in a composition for activating cytochrome c
FR2925326A1 (en) * 2007-12-21 2009-06-26 Vincience Sa Use of maize (Zea mays L.) peptide hydrolysate, as active ingredient to activate the synthesis of aquaporins with other active ingredients, and in cosmetic or nutraceutical composition to improve hydration and barrier function of skin
FR2925325A1 (en) * 2007-12-21 2009-06-26 Vincience Sa Use of rapeseed (Brassica napus) peptide hydrolysate, as active ingredient to activate the synthesis of aquaporins with other active ingredients, and in cosmetic or nutraceutical composition to improve hydration and barrier function of skin
WO2010029115A1 (en) * 2008-09-10 2010-03-18 L V M H Recherche Methods useful in studying or modulating skin or hair pigmentation, plant extracts for use in compositions and cosmetic care method
US9668960B2 (en) 2008-09-10 2017-06-06 L V M H Recherche Methods useful in studying or modulating skin or hair pigmentation, plant extracts for use in compositions and cosmetic care method
KR101494375B1 (en) * 2008-09-10 2015-02-17 엘브이엠에이취 러쉐르쉐 Methods useful in studying or modulating skin or hair pigmentation, plant extracts for use in compositions and cosmetic care method
US8409633B2 (en) 2008-09-10 2013-04-02 L V M H Recherche Methods useful in studying or modulating skin or hair pigmentation, plant extracts for use in compositions and cosmetic care method
EP2260831A1 (en) * 2009-06-12 2010-12-15 L V M H Recherche Plant extracts modulating Myo-X for use in compositions
FR2956818A1 (en) * 2010-02-26 2011-09-02 Isp Investments Inc Cosmetic use of peptide hydrolyzate from flax (Linum kind), as active agent in composition to soothe the skin, to prevent the damage or restore the skin barrier function and for preventing or treating skin inflammations e.g. reddening
WO2014029948A2 (en) * 2012-08-24 2014-02-27 Isp Investments Inc. Flax extract and cosmetic composition comprising said extract for increasing the level of intracellular coenzyme q10
FR2994656A1 (en) * 2012-08-24 2014-02-28 Isp Investments Inc FLAX EXTRACT AND COSMETIC COMPOSITION COMPRISING SAID EXTRACT FOR INCREASING THE INTRACELLULAR Q10 COENZYME LEVEL
WO2014029948A3 (en) * 2012-08-24 2014-07-10 Isp Investments Inc. Flax extract and cosmetic composition comprising said extract for increasing the level of intracellular coenzyme q10
WO2014090526A3 (en) * 2012-12-12 2014-12-11 Henkel Ag & Co. Kgaa Active substance combination for skin lightening i
ITBO20130279A1 (en) * 2013-05-31 2014-12-01 Phenbiox S R L COMPOSITION FOR BODY CARE

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