EP2150232A2 - Verwendung eines aus amaranth gewonnenen wirkstoffbestandteils zur herstellung einer zusammensetzung zur aktivierung einer zellenergie und zum schutz der haut gegen oxidative beschädigung - Google Patents

Verwendung eines aus amaranth gewonnenen wirkstoffbestandteils zur herstellung einer zusammensetzung zur aktivierung einer zellenergie und zum schutz der haut gegen oxidative beschädigung

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Publication number
EP2150232A2
EP2150232A2 EP08805495A EP08805495A EP2150232A2 EP 2150232 A2 EP2150232 A2 EP 2150232A2 EP 08805495 A EP08805495 A EP 08805495A EP 08805495 A EP08805495 A EP 08805495A EP 2150232 A2 EP2150232 A2 EP 2150232A2
Authority
EP
European Patent Office
Prior art keywords
active ingredient
skin
composition
effective amount
active
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP08805495A
Other languages
English (en)
French (fr)
Inventor
Claude Dal Farra
Nouha Domloge
Jean-Marie Botto
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Ashland Specialties France
Original Assignee
Societe dExtraction des Principes Actifs SA
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Filing date
Publication date
Application filed by Societe dExtraction des Principes Actifs SA filed Critical Societe dExtraction des Principes Actifs SA
Publication of EP2150232A2 publication Critical patent/EP2150232A2/de
Withdrawn legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/482—Cassia, e.g. golden shower tree
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/68—Plantaginaceae (Plantain Family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/01—Hydrolysed proteins; Derivatives thereof
    • A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics 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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier 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
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations

Definitions

  • the present invention is in the cosmetic and pharmaceutical field, and more particularly in the field of dermatology.
  • the present invention relates to the use in a cosmetic composition or for the preparation of a pharmaceutical composition, of an effective amount of a peptide active ingredient derived from amaranth of the species Amaranthus hypochon ⁇ iacus; said active ingredient, or the composition containing it, being intended to activate cellular energy and protect the skin from oxidative damage.
  • the active ingredient can be used alone or in combination with at least one other active ingredient.
  • the invention still bears. on a cosmetic treatment process intended to protect the skin and the integuments from external aggressions and to fight against skin aging.
  • Said active ingredient, activator of cellular energy and skin protector can also be used to prepare pharmaceutical compositions intended to prevent or fight against pathologies related to oxidation processes, or certain pathologies of aging.
  • “superficial body growths” encompasses all the keratinous appendages present on the surface of the body, in particular the hairs, the eyelashes, the eyebrows, the nails and the hair.
  • the skin is a vital organ that covers the entire surface of the body and provides protective, sensitive, immune, metabolic or thermoregulatory functions.
  • the skin like other organs, is subject to aging.
  • one of the major mechanisms involved in aging processes is the accumulation of oxidative damage in essential molecules such as membrane lipids, proteins, DNA and especially mitochondrial DNA (mtDNA).
  • Oxidative damage is caused by free radicals, chemically unstable and highly reactive species, generated by intracellular metabolism or external aggression. These external aggressions include: UV radiation, toxins, air pollutants, food oxidants. In the skin, there is premature aging occurring in the areas exposed to radiation, characterized by phenomena of alterations of macromolecules (lipid peroxidation, carbonylation of proteins) affecting in particular elastin, collagen or fibronectin. It has also been possible to show a progressive decline of mitochondrial functions with age, .
  • the body has defense mechanisms, capable of trapping or transforming free radicals (enzymes, glutathione, vitamins A and E, coenzyme Q10, etc.).
  • these systems of antioxidant defenses often prove insufficient in the face of the many stresses and external aggressions to which organisms and the skin in particular are subjected.
  • Coenzyme Q10 (or ubiquinone) is a coenzyme found in mitochondrial complexes involved in oxidative phosphorylation leading to the production of ATP (Mitchell et al, 1976, Mitchell et al, 1990).
  • the other fundamental property of coenzyme Q10 is to be an antioxidant, neutralizing free radicals (Beyer et al 1990,
  • Coenzyme Q10 is a benzoquinone derivative flanked by a long isoprene side chain usually composed of ten isoprenoid units (hence the name Coenzyme Q10). Since this coenzyme is not soluble in water, it is found only in lipid membranes, such as the inner membrane of the mitochondria, where it can diffuse freely among membrane phospholipids.
  • Coenzyme Q10 can exist under three oxidation states: a reduced form (CoQH2 or UQH2), an oxidized form (CoQ10), and an intermediate form the radical ubisemiquinone (Q 0 ) - Coenzyme Q10 is present in the skin where it stimulates the natural functions of cells and acts as a defender vis-à-vis external aggressions.
  • a reduced form CoQH2 or UQH2
  • CoQ10 oxidized form
  • Q 0 radical ubisemiquinone
  • coenzyme Q10 The biosynthesis of coenzyme Q10 is made from tyrosine for the quinone nucleus, and from farnesyl pyrophosphate for the side chain.
  • the enzyme responsible for this latter reaction which is an essential step in the biosynthesis of coenzyme Q10, is transprenyl transferase (or polyprenyl transferase).
  • the main subject of the present invention is the use of a peptide active principle resulting from the hydrolysis of Amaranthus hypochondriacus amaranth, able to protect the skin from external aggressions and to fight against skin aging.
  • Said active ingredient may be used alone or in combination with at least one other active ingredient.
  • the inventors have demonstrated a therapeutic activity, and more particularly dermatological and cosmetic, of such an active ingredient.
  • this active ingredient when applied to the skin, has a high protective activity with respect to the oxidative damage to the skin and significantly promotes the synthesis of ATP, as well as that the synthesis or activity of the enzyme transprenyl transferase and coenzyme Q10.
  • active ingredient capable of increasing cellular energy and protecting the skin from oxidative damage means any substance of plant origin, and more particularly from amaranth of the species Amaranthus hypochondriacus capable of increasing the synthesis of intracellular ATP and to exhibit protective properties in cells or tissues subjected to oxidative stress of physicochemical or environmental origin.
  • the active ingredient capable of increasing cellular energy and protecting the skin from oxidative damage is, according to the invention, an extract of a peptide nature and comes from the hydrolysis of Amaranthus amaranth proteins. hypochondriacus.
  • eptide is understood to mean a mixture of compounds predominantly represented by peptides or polypeptides.
  • peptide refers to a sequence of two or more amino acids linked together by peptide bonds or modified peptide bonds.
  • biologically active is meant “which has an activity in vivo or in vitro characteristic of the activity of the active ingredient according to the invention”.
  • hydrolyzate or derived from hydrolysis refers to any substance or mixture of substances, or isolated preparation, obtained after hydrolysis of plant material.
  • the peptide active principle according to the invention is obtained by extraction of proteins of plant origin, followed by controlled hydrolysis which releases biologically active peptide fragments.
  • Amaranth is an annual plant of the family Amaranthaceae belonging to the genus Amaranthus, some of which are grown as ornamental plants for their spectacular flowering spikes, and sometimes as vegetables, for their spinach-like edible leaves or for their seeds.
  • the species used is Amaranthus hypochondriacus
  • the plant material used is the seed and preferably the seed removed from its shell by a dehulling step.
  • the plant is ground using a plant grinder.
  • the powder thus obtained may subsequently be "delipidated” with the aid of a conventional organic solvent (for example an alcohol, hexane or acetone).
  • a conventional organic solvent for example an alcohol, hexane or acetone.
  • the proteins of the plant are then extracted according to the conventional method (Osborne, 1924) modified; the plant meal is suspended in an alkaline solution containing an insoluble polyvinylpolypyrrolidone (PVPP) adsorbent product (0.01 - 20%); in fact, it has been observed that the hydrolysis and subsequent purification operations are facilitated by this means. The concentration of phenolic-type substances interacting with the proteins is thus reduced.
  • PVPP polyvinylpolypyrrolidone
  • 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.
  • the proteins are then precipitated by varying the ionic strength by acidifying the medium, which eliminates soluble components and nucleic acids.
  • the precipitate is then washed with an organic 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 other solvent and is then a more purified form of the hydrolyzate.
  • the extraction can also be carried out in neutral or acidic medium always in the presence of polyvinylpolypyrrolidone.
  • the precipitation step is then carried out using a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate) or an organic solvent (alcohol, acetone).
  • a conventional precipitation agent such as salts (sodium chloride, ammonium sulfate) or an organic solvent (alcohol, acetone).
  • the precipitate obtained can be separated from the precipitating agents by dialysis after redissolving in water or another solvent.
  • 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 of plant origin papain, bromelain, ficin
  • microorganisms Aspergillus, Rhizopus, Bacillus, etc.
  • the active hydrolyzate can be further purified in order to select the molecular weights and the nature of the peptides generated. Splitting can - -
  • any of the more or less purified forms of the hydrolyzate is then solubilized in water or in any mixture containing water, and then sterilized by ultrafiltration.
  • the vegetable hydrolyzate obtained according to the invention is analyzed qualitatively and quantitatively for its physico-chemical characteristics and its content of protein and peptide compounds.
  • Peptide-like 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.
  • the active plant hydrolyzate has a pH of between 4 and 7, and preferably between 5 and 6, a solids content of between 1 and 8 g / l, and preferably between 2 and 5 g / l, its content of peptide compounds is between 0.1 and 5 g / l, and preferably between 0.5 and 2 g / l, and its sugar content is 0.5 to 2 , 5 g / 1.
  • the peptide active ingredient according to the invention is first solubilized in one or more solvents conventionally used by those skilled in the art, such as water, glycerol, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols, petroleum jelly, a vegetable oil or any mixture of these solvents.
  • solvents conventionally used by those skilled in the art, such as water, glycerol, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols, petroleum jelly, a vegetable oil or any mixture of these solvents.
  • the active principle according to the invention is solubilized beforehand 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.
  • 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 to the skin.
  • These compositions must therefore contain a cosmetically and / or dermatologically acceptable medium, that is to say compatible with the skin and superficial body growths, and cover all cosmetic or dermatological forms.
  • These compositions may especially be in the form of creams, oil-in-water or water-in-oil emulsions, multiple emulsions, solutions, suspensions, gels, milks, lotions, sticks or powders, suitable for application to the skin, lips and / or integuments.
  • compositions comprise the excipients necessary for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes.
  • solvents such as solvents, thickeners, diluents, surfactants, antioxidants, dyes, preservatives, perfumes.
  • 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.
  • compositions that can be used also contain at least one other active ingredient promoting the action of the peptide active principle according to the invention.
  • the composition according to the invention may combine, with the active ingredient capable of increasing cellular energy and protecting the skin from oxidative damage, active ingredients having an antioxidant action, or stimulating the synthesis of dermal macromolecules, or stimulating energy metabolism.
  • active ingredient having an anti-radical or antioxidant action mention may be made of vitamin C, vitamin E or coenzyme Q10 or polyphenolic extracts of plants.
  • anti-radical active ingredients is meant any compound capable of trapping free radicals. These active principles are capable of blocking free radical chain reactions before the ultimate stages of degradation of the biological constituents of the skin and thus have an antioxidant activity.
  • the composition according to the invention may be a solar composition, that is to say a composition helping to protect against solar radiation. - -
  • active assisting solar protection such as, for example, sunscreens.
  • the effective amount of active ingredient corresponds to the amount of amaranth hydrolyzate obtained according to the invention, necessary to obtain the desired result, namely: to increase the synthesis of ATP, to protect the skin from oxidative damage and more generally to protect the skin. skin external aggression and prevent or treat skin aging.
  • the active principle derived from amaranth is present in the compositions of the invention at a concentration of from 0.0001% to about 20%, and preferentially at a concentration of between 0, Approximately 5% and 5% relative to the total weight of the final composition.
  • compositions may especially be in the form of an aqueous solution, hydroalcoholic or oily; an oil-in-water, water-in-oil emulsion or multiple emulsions; they may also be in the form of creams, suspensions or powders, suitable for application to the skin, mucous membranes, lips and / or integuments.
  • These compositions may be more or less fluid and have the appearance of a cream, lotion, milk, serum, ointment, gel, paste or paste. a foam. They can also be in solid form, as a stick, or be applied to the skin in the form of an aerosol. They can be used as a care product and / or as a make-up product for the skin.
  • compositions additionally comprise any additive commonly used in the field of application envisaged as well as the adjuvants necessary for their formulation, such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning principles, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc. .
  • additives such as solvents, thickeners, diluents, antioxidants, dyes, sunscreens, self-tanning principles, pigments, fillers, preservatives, perfumes, odor absorbers, cosmetic or pharmaceutical active ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.
  • these adjuvants and their proportions are chosen in such a way as not to harm the desirable advantageous properties of the composition according to the invention.
  • These adjuvants may, for example, correspond to 0.01 to 20% of the total weight of the composition.
  • the fatty phase may represent from 5 to 80% by weight and preferably from 5 to 50% by weight relative to the total weight of the composition.
  • the emulsifiers and co-emulsifiers used in the composition will be chosen from those conventionally used in the field under consideration. For example, they can be used in a proportion ranging from 0.3 to 30% by weight, relative to the total weight of the composition.
  • the active ingredient according to the invention can be used advantageously in a cosmetic composition or for the preparation of a pharmaceutical composition.
  • the active ingredient according to the invention may advantageously be used in a cosmetic composition to fight in a preventive and / or curative manner against the manifestations of cutaneous aging and, more specifically, in order to fight against and / or prevent aging.
  • -induced (photo-aging) Skin manifestations of aging means any changes in the external appearance of the skin due to aging such as, for example, wrinkles and fine lines, wilted skin, soft skin, thinned skin, lack of elasticity and / or or skin tone, dull and lackluster skin or skin pigmentation spots, but also any internal changes in the skin that do not systematically result in a modified external appearance such as, for example, any internal degradation of the skin. skin following exposure to ultraviolet (UV) radiation.
  • UV ultraviolet
  • the active ingredient according to the invention will make it possible, in particular, to combat the loss of elasticity and firmness of the skin.
  • the use of the active ingredient alone, or a composition containing it, will allow the skin and integuments to be protected and to better withstand environmental stresses.
  • an essential aspect of the invention is the use of the active principle according to the invention, in a cosmetic composition for the protection of the skin and integuments against oxidative damage by virtue of a protective activity with regard to the species reactive oxygen.
  • Said active ingredient being advantageously used as an antioxidant active ingredient, and / or as an anti-radical active ingredient, and / or as an anti-glycation active ingredient.
  • anti-radical active principle any compound capable of trapping free radicals before the ultimate stages of degradation of the biological constituents of the skin, so-called antioxidant compounds.
  • anti-glycation active principle any compound capable of limiting the cellular damage caused by glycation or glycoxidation reactions.
  • the active ingredient according to the invention will combat the aesthetic damage caused to the skin and / or hair by free radicals.
  • the active agent can be used advantageously in a cosmetic composition for the protection of the skin and integuments against all types of external aggression.
  • external aggression refers to the aggressions that the environment can produce. By way of example, mention may be made of aggressions such as pollution, UV, or irritating products such as surfactants, preservatives or perfumes.
  • Pollution is understood to mean both “external” pollution, due for example to diesel particles, ozone or heavy metals, and “internal” pollution, which may be due in particular to the solvent emissions of paints, glues , or wallpaper (such as toluene, styrene, xylene or benzaldehyde), or even cigarette smoke.
  • the active ingredient according to the invention may advantageously be used in a cosmetic composition or for the preparation of a pharmaceutical composition, as a photo-protective active ingredient and, more particularly, as a so-called “secondary" photo-protective active ingredient. .
  • the primary photoprotective active principles are distinguished from the secondary photoprotective active ingredients.
  • Primary photoprotective active ingredients are substances that exercise physical power: they are able to absorb UV radiation and return it as heat to protect the skin.
  • Secondary photoprotective active ingredients are substances that usually have a biological effect; these are, for example, the active ingredients capable of limiting damage to DNA and membranes by the penetration of UV radiation into the skin.
  • the subject of the invention is also the use in a cosmetic composition of an effective amount of active ingredient as described above, for preventing damage to the skin caused by exposure to the sun or exposure to ionizing radiation during radiotherapy.
  • the subject of the invention is also the use in a cosmetic composition of an effective amount of active principle as described above, for increasing the synthesis of intracellular ATP of the cells of the skin.
  • the subject of the invention is also the use in a cosmetic composition of an effective amount of active principle as described above, for increasing the activity or the synthesis of the enzyme transprenyl transferase and / or coenzyme Q10, in the cells of the skin.
  • the invention also relates to the use in a cosmetic composition of an effective amount of active ingredient as described above to protect the skin from damage caused by free radicals.
  • the invention also consists in the use of an effective amount of active principle as described above, for the manufacture of a pharmaceutical composition intended to attenuate a pathology related to oxidation processes, or certain pathologies of aging.
  • the invention also consists of a cosmetic treatment method intended to protect the skin and the integuments from external aggressions and to fight against skin aging characterized by the application on the skin or integuments to be treated of a composition containing an effective amount of the active ingredient according to the invention.
  • the active agent is obtained from an extract of plants of the species Amaranthus hypochondriacus.
  • a first step 1 kg of shelled amaranth seeds are crushed in a cereal mill and the flour obtained is delipidated by the action of an organic solvent: hexane.
  • the powder obtained is suspended in an aqueous alkaline solution (dilution 1/10) pH 10 containing 1% polyvinylpolypyrrolidone (Polyclar V ISP). 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 amaranth, is then subjected to protein precipitation by varying the ionic strength in neutral or acidic medium, thereby eliminating soluble carbohydrate components, lipids and nucleic acids.
  • the medium is brought to pH 3.5.
  • 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 solvent such as, for example, ethanol or methanol
  • the protein-rich precipitate is dissolved in water or another solvent.
  • the crude protein extract is then subjected to a series of gentle and selective hydrolyses consisting of chemical and enzymatic hydrolyses in the presence of 0.5% PVPP (Polyclar V) and cysteine endopeptidases (papain, ficin). After reaction, the hydrolyzate is filtered on a plate and then on a sterilizing cartridge (0.2 ⁇ m).
  • PVPP Polyclar V
  • cysteine endopeptidases papain, ficin
  • a light-colored hydrolyzate of 15 to 30 g / l of dry extract is then obtained, which is then diluted so that the concentration of peptides determined by the method of Lowry is between 0.1 and 5 g. and preferably between 0.5 and 2 g / l.
  • the physicochemical analysis of the vegetable hydrolyzate, which constitutes the active ingredient shows that its pH is between 4 and 7, and preferably between 5 and 6, the dry extract is between 1 to 8 g / l and preferably between 2 and 5 g / 1, its content of peptide compounds is between 0.1 and 5 g / l and preferably between 0.5 to 2 g / 1 and its sugar content between 0.5 to 2, 5 g / 1.
  • a variant of the protocol of Example 1 consists in carrying out the same sequence of controlled and selective enzymatic hydrolyses but in the presence of 0.5% of PVPP.
  • a light-colored hydrolyzate of 15 to 30 g / l of dry extract is obtained after sterilizing filtration.
  • the solution is then ultrafiltered on a Millipore Helicon filter cartridge (eut off 1 kD).
  • the high molecular weights contained in the retentate are removed, the filtrate is preserved.
  • the concentration of compounds of peptide nature is determined by the method of Lowry, between 0.1 and 5 g / l and preferably between 0.5 and 2 g / l.
  • Another variant consists in carrying out a purification of the active principle, obtained according to Example 1 or 2, by ion exchange chromatography, on a TSK gel column (TosoHaas) with a pH 7 phosphate buffer.
  • the purpose of this study is to determine the influence of the active ingredient according to Example 1 on the synthesis of ATP.
  • the dermal fibroblasts are treated with a solution containing 1% of active principle according to Example 1, for a period ranging from 1 to 3 hours.
  • the wells are emptied of their medium and rinsed with 2 ml of cold PBS before adding 250 ⁇ l of a lysis buffer provided by the kit.
  • the cells of each well are then scraped and then harvested in 14 ml tubes.
  • Each well is rinsed with 2 x 500 ⁇ l of cold PBS and the whole is again harvested in the respective tubes. From these samples, dilution is carried out at 1/12000 in cold PBS before each reading.
  • the ATP assay is performed on these samples: 50 ⁇ L of this dilution are deposited in a lumen and 50 ⁇ L of luminol are added. After 10 seconds, the reading of the luminescence is triggered. The values are standardized with respect to the amount of protein for each sample. Measurements are made with one device: the LUMAC® / 3M Biocounter M2010A. Results
  • the ATP assays show that there is an increase in the amount of intracellular ATP of 11% after 1 hour and 67% after 3 hours of culture in cells treated with the active ingredient according to Example 1, in comparison of untreated cells.
  • the active ingredient according to Example 1 strongly activates the synthesis of intracellular ATP in cutaneous cells.
  • Example 1 vis-à-vis dermal fibroblasts subjected to oxidative stress caused by UVB radiation or by hydrogen peroxide (H 2 O 2 ).
  • H 2 O 2 hydrogen peroxide
  • protein carbonylation assays were performed. The carbonylation of proteins results from the oxidative cleavage of proteins or from the oxidation of arginine, lysine, proline or threonine residues. The determination of protein carbonylation is carried out by an EIA (Enzyme Immuno Assay) technique.
  • EIA Enzyme Immuno Assay
  • Fibroblasts in culture were placed in the presence of the active ingredient according to Example 1 at 1%, 72 hours before, during and 24 hours after the oxidative stress (UVB irradiation at 50 mJ / cm 2 or treatment with 2 mM of H 2 O 2 ). Untreated and non-oxidative stress controls are performed.
  • the measurement of carbonylation consists in using DNP (dinitrophenyl) which has the property of binding specifically on the carbonyl groups of the proteins.
  • DNP dinitrophenyl
  • the fixed DNP will then be assayed by an ELISA method, thanks to an anti-DNP antibody coupled to a peroxidase.
  • a range of oxidized BSA (bovine serum albumin) (whose concentration in carbonyl groups is known) is used for calibration.
  • the results obtained show a 30% decrease in the carbonylation of the proteins when the cells are treated with the active principle according to Example 1 according to the invention, compared with the untreated cells.
  • the active ingredient according to Example 1 effectively protects the skin cells against oxidative damage caused by UVB radiation or hydrogen peroxide.
  • the purpose of this study is to determine the protective effect of the active ingredient according to Example 1, vis-à-vis ex vivo epidermis culture subjected to stress by a glycating agent.
  • Human skin biopsies are maintained in culture ex vivo, treated with a 1% solution, 24 hours before, and another 24 hours after the contact with a glycogen agent (methyl glyoxal 5 or 10 mM).
  • a glycogen agent methyl glyoxal 5 or 10 mM.
  • Hematoxylin-Eosin (H & E) histological sections and stainings are used to evaluate the quality of cutaneous structures.
  • the active ingredient according to Example 1 protects the skin from a stress induced by glycation.
  • phase A and phase B are separately heated between 70 ° C. and 75 ° C.
  • Phase B is emulsified in phase A with stirring.
  • Phase C is added at 45 ° C, increasing stirring.
  • Phase D is then added when the temperature is below 40 ° C. Cooling is continued up to 25 ° C. with vigorous stirring.
  • phase A Prepares phase A with stirring. Incorporate the xanthan gum gradually, with deflocculating stirring. Phases C and D will be incorporated once the gel is complete. Phase E, prepared before perfect dissolution of the DHA, will be added later. Adjust the pH if necessary to 4 - 4.5. Color and perfume.
  • phase A Prepare and melt phase A at 65-70 ° C. Heat phase C at 65-70 ° C. Phase B is added to phase A just before emulsifying A in B. At about 45 ° C, the carbomer is neutralized by addition of phase D. Phase E is then added with gentle stirring and cooling is continued to 25 ° C. Phase F is then added if desired.
  • phase A Prepare phase A and heat to 75 ° C with stirring.
  • Prepare phase B by dispersing the carbopol, then the xanthan gum with stirring. Let rest. Heat at 75 ° C. At temperature, emulsify A in B with rotor-stator stirring. Neutralize with phase C with rapid stirring. After cooling to 40 ° C., add the phase D and then the phase E. The cooling is continued with gentle stirring and the F phase added.

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EP08805495A 2007-04-27 2008-04-23 Verwendung eines aus amaranth gewonnenen wirkstoffbestandteils zur herstellung einer zusammensetzung zur aktivierung einer zellenergie und zum schutz der haut gegen oxidative beschädigung Withdrawn EP2150232A2 (de)

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FR0703062A FR2915385B1 (fr) 2007-04-27 2007-04-27 Utilisation d'un principe actif issu de l'amarante (amaranthus) pour preparer une composition destinee a activer l'energie cellulaire et a proteger la peau des dommages oxydatifs.
PCT/FR2008/000576 WO2008145853A2 (fr) 2007-04-27 2008-04-23 Utlisation d'un principe actif issu de l'amarante (amaranthus) pour preparer une composition destinee a activer l'energie cellulaire et a proteger la peau des dommages oxydatifs

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EP2150232A2 true EP2150232A2 (de) 2010-02-10

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FR2948876B1 (fr) * 2009-08-07 2011-11-04 Oreal Association d'un rayonnement lumineux et d'un agent de nature peptidique augmentant l'expression d'un substrat de la cytochrome c oxydase pour ameliorer notamment l'apparence de la peau et/ou du cheveu
WO2011124861A2 (fr) * 2010-04-09 2011-10-13 Greentech Utilisation de composés oligosaccharides dérivés du xanthane comme agent énergisant
FR2958648B1 (fr) * 2010-04-09 2015-10-30 Greentech Nouveaux composes oligosaccharides et utilisation comme agent energisant
JP2023103165A (ja) * 2022-01-13 2023-07-26 株式会社日清製粉グループ本社 Ａｔｐ産生促進剤、抗炎症剤及び飲食品

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US5670635A (en) * 1993-11-29 1997-09-23 Datta; Asis Seed storage protein with nutritionally balanced amino acid composition
EP1382326A1 (de) * 2002-07-17 2004-01-21 Cognis France S.A. Verfahren zum Schutz der Haut gegen die Alterung
WO2004043482A1 (fr) * 2002-11-08 2004-05-27 Societe D'extraction Des Principes Actifs (Vincience) Composition cosmetique ou pharmaceutique comprenant des peptides, possedant le motif arg-gly-ser
DE10307116A1 (de) * 2003-02-19 2004-09-02 Hans Schwarzkopf & Henkel Gmbh & Co. Kg Haarbehandlungsmittel mit neuartiger Wirkstoffkombination
RU2266750C2 (ru) * 2003-07-31 2005-12-27 Одинец Алексей Глебович Способ получения биологически активной сыворотки, обогащенной пептидами
WO2005097060A1 (fr) * 2004-03-12 2005-10-20 Societe D'extraction Des Principes Actifs Sa (Vincience) Utilisation de peptides en tant qu’agent antioxydant pour la preparation d’ une composition cosmetique et/ou pharmaceutique
US7399491B2 (en) * 2004-12-28 2008-07-15 Council Of Scientific And Industrial Research Health promoting functional foods fortified with herbs

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WO2008145853A2 (fr) 2008-12-04
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FR2915385A1 (fr) 2008-10-31
WO2008145853A3 (fr) 2009-03-05

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