WO2023237833A1 - Anti-ageing composition with rose extracts and peptides - Google Patents

Anti-ageing composition with rose extracts and peptides Download PDF

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Publication number
WO2023237833A1
WO2023237833A1 PCT/FR2023/050797 FR2023050797W WO2023237833A1 WO 2023237833 A1 WO2023237833 A1 WO 2023237833A1 FR 2023050797 W FR2023050797 W FR 2023050797W WO 2023237833 A1 WO2023237833 A1 WO 2023237833A1
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Prior art keywords
rosewood
composition
tetrapeptide
skin
evanrat
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PCT/FR2023/050797
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French (fr)
Inventor
Jocelyne Franchi
Carine Nizard
Valérie GORZELANCZYK
Laure VERT
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L V M H Recherche
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Publication of WO2023237833A1 publication Critical patent/WO2023237833A1/en

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    • 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
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations

Definitions

  • the present invention relates to a cosmetic composition, in particular a cosmetic care composition intended to prevent and/or reduce the signs of skin aging, in particular of the skin of the face and/or body.
  • the skin is the body's first protective barrier against the environment. It is daily subject to the effects of factors of exogenous origin (e.g.: UV radiation, temperature variations, atmospheric pollution, cigarette smoke, etc.) and/or endogenous (e.g.: hormones, etc.).
  • exogenous origin e.g.: UV radiation, temperature variations, atmospheric pollution, cigarette smoke, etc.
  • endogenous e.g.: hormones, etc.
  • Aging leads to changes in the skin of the body and/or face, including the lips, areas particularly subject to the deleterious effects of the environment, such as wrinkles and fine lines, skin having lost its firmness, suppleness and its elasticity, a reduction in the thickness of the epidermis linked in particular to a slowdown in the renewal of skin cells.
  • a large number of anti-aging active ingredients such as peptides, vitamins, antioxidants and plant extracts are known from the prior art. Mention may in particular be made of the peptides Acetyl Hexapeptide-8 from the company Lipotec and Palmitoyl Tetrapeptide-7 from the company Sederma.
  • the Applicant has demonstrated a synergistic anti-aging in vitro effect between the peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7) and an aqueous extract of rosewood of the Evanrat variety. It also demonstrated anti-aging synergy between peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7), an extract aqueous rosewood of the Evanrat variety and a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety.
  • the present invention therefore relates in particular to a cosmetic composition
  • a cosmetic composition comprising, in a physiologically acceptable medium:
  • the present invention also relates to a non-therapeutic cosmetic process for caring for and/or making up the skin and/or lips comprising the application to said skin and/or said lips, in particular the skin, of a cosmetic composition such as as defined in the present invention.
  • the invention also relates to a combination of cosmetic active ingredients comprising:
  • an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
  • the invention further relates to the non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined in the present invention, as an active association for preventing and/or reducing the signs of skin aging, in particular loss of density. and/or firmness of the skin, reduction in the thickness of the epidermis, reduction differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines.
  • the invention also relates to a cosmetic set comprising:
  • an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium, characterized in that at least one of the active ingredients (i) to (iv) is packaged in a composition distinct from the other active ingredients, and in particular each active ingredient is packaged in a distinct composition.
  • the invention finally relates to a cosmetic process comprising the successive application, to the skin and/or lips, of the compositions of the cosmetic assembly as defined in the present invention.
  • FIG. 1 Effect of the different active ingredients and combinations of active ingredients according to the invention on the protein expression of the KI67 factor in cultures of reconstructed human skin.
  • FIG. 2 Effect of the different active ingredients and combinations of active ingredients according to the invention on the thickness of the epidermis in cultures of reconstructed human skin.
  • FIG. 3 Effect of the different active ingredients and combinations of active ingredients according to the invention on the protein expression of Collagen VII in cultures of reconstructed human skin.
  • the present invention therefore relates in particular to a cosmetic composition
  • a cosmetic composition comprising, in a physiologically acceptable medium: (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
  • the roses or rose bushes used as plant material for the preparation of the extracts used according to the invention correspond to the rose bush or rose 'Jardin de Granville®', a hybrid variety offered exclusively by "Roses ancient André Eve S.A.S” and protected by Certificate of Obtention Plant under number 20110345 with the species name Rosa L. and the variety the name EVANRAT.
  • This bush rose belongs to the group of modern hybrids, which, from May to October, is permanently covered with roses, thus showing an excellent repeating character.
  • the aqueous extract of rosewood of the Evanrat variety or 'Jardin de Granville®' rose, preferably summer rosewood, used according to the invention is obtained by enzymatic extraction using a cosmetically acceptable polar solvent. , according to the techniques described in application FR3090380.
  • polar solvent means that the solvent has a Polarity index value which is equal to or greater than a value of 4.
  • the polarity index is a quantity calculated on the basis of thermodynamic quantities (solubility and change of state) which highlights the more or less polar nature of a molecule. Refer, for the polarity indices of solvents, to the article by L.R. SNYDER: Classification of the solvent properties of common liquids; Journal of Chromatography, 92 (1974), 223-230.
  • the preferred polar solvents are those consisting of a compound comprising at least one polar covalent bond of the OH type.
  • a cosmetically acceptable polar solvent a solvent or a mixture of solvents chosen from water, C1-C4 alcohols, such as ethanol, glycols, such as ethylene glycol, glycerol, butylene glycol and propylene glycol, and mixtures thereof.
  • water is used.
  • said extract is characterized in that it comprises water in a content ranging from 94 to 96%, a dry extract content ranging from 3 to 5% and preservatives in a content ranging from 0, 5 to 1% by weight relative to the total weight of the extract.
  • the rosewood according to the invention is obtained from freshly cut whole roses and is isolated from the rest of the plant by manual or mechanical separation.
  • the rosewoods are directly used for extraction.
  • the rosewood is previously dried before extraction.
  • the rosewood is crushed before the extraction stage.
  • Rosewood includes monosaccharide sugars (fructose, glucose), and polysaccharides in particular disaccharides, polyphenols (catechin), amino acids (mainly aspartic acid, tyrosine and arginine), minerals (ash, sodium, potassium, calcium), flavonoids (mostly glycosylated) and tannins.
  • monosaccharide sugars fructtose, glucose
  • polysaccharides in particular disaccharides
  • polyphenols catechin
  • amino acids mainly aspartic acid, tyrosine and arginine
  • minerals ash, sodium, potassium, calcium
  • flavonoids mostly glycosylated
  • the aqueous rosewood extract according to the invention is an extract concentrated in polar natural compounds.
  • the aqueous extract of rosewood according to the invention is distinct from rose water. This is a rosewood extract using the extraction of natural rosewood compounds in the presence of aqueous solvents (polar) with or without the addition of a pH modifier, and an implementation, in the formulation, in the form of an aqueous solution or concentrate, the solvent of the concentrate possibly being the extraction solvent and/or an additional solvent.
  • the aqueous extract of rosewood according to the invention further comprises in a small proportion an extract of rose flowers representing from 0.01 to 1%, preferably from 0.05 to 0.5%, preferably 0.1 to 0.2% of the total aqueous rose extract according to the invention.
  • the original plant material comprises rosewood and rose flowers in a proportion 0.01% flowers for 99.99% wood to 1% flowers for 99% wood.
  • the aqueous extract of rosewood according to the invention can be obtained according to different eco-extraction processes known to those skilled in the art and in particular that described below.
  • rosewood of the Evanrat variety preferably the Jardin de Granville® rose, preferably wood of roses previously dried.
  • the wood can be used fresh, frozen or previously dried using conventional drying tools known to those skilled in the art, such as drying in the open air, in ovens, by freeze-drying or zeodration.
  • drying in the open air, in ovens by freeze-drying or zeodration.
  • the wood may have been dried and/or crushed.
  • the wood is used wet then crushed.
  • the wood is crushed before extraction, for example by means of a mortar, cryo-grinding, a mixer, a traditional grinder or a centrifuge according to methods known to those skilled in the art. job.
  • the aqueous extract of rosewood according to the invention can be obtained in particular according to enzymatic extraction processes.
  • the enzymes will be adapted to the walls to be hydrolyzed to access these said molecules.
  • a cellulase to release glucose and cellulose; a hemicellulase to release reduced sugar mono- and oligomers; a pectinase to release uronic acids; and advantageously a protease to hydrolyze the structural proteins and improve the lysis of the raw material.
  • eco-extraction is carried out using the protocol described in patent application WO2011045387 (corresponding to patent application FR2351461) in the name of the National Polytechnic Institute of Lorraine INPL.
  • This is an alternative technology that meets regulatory and environmental constraints while remaining economically competitive.
  • This innovative process makes it possible to extract from plant material, in a single step, three types of products, namely an oil (rich in polyphenol nutrients, sterols, vitamin E, etc.), an aqueous extract as well as a cake ( solid phase).
  • the aqueous extract of rosewood used according to the invention is obtained by an enzymatic extraction process comprising the steps comprising the steps: a) addition of water to the wood of previously crushed rose having an appropriate particle size, b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously also at least one protease, c) incubation with stirring of the wood of previously crushed rose and the enzymatic mixture to release into the reaction medium oils, proteins and fermentable sugars, for a period depending on the desired yields, d) separation of the reaction medium to obtain free oil, an aqueous phase containing fermentable proteins and sugars, and a solid phase, and e) separation of the aqueous phase and possible addition of preservatives.
  • step a) the appropriate particle size of the plant material is advantageously obtained by grinding said plant material.
  • the grinding must be as fine as possible to promote the action of the enzymes. Ideally, all particles should be close to 50 pm in size, preferably close to 10 pm.
  • the mass of water added to the plant material is equal to 1 to 2 times the mass of said plant material and does not exceed this quantity.
  • step b) the ratio between the pectinase activity and the cellulase activity being at least 0.14, preferably between 0.3 and 2.5, and more preferably between 0.35 and 0.45, and the ratio between the pectinase activity and the hemicellulase activity being at least 7.10 -3 ; preferably between 1.10 -2 and 0.5, and more preferably between 1.10 -2 and 2.10 -2 .
  • the enzymatic mixture used contains at least one cellulase, at least one hemicellulase, at least one pectinase, and advantageously also at least one protease.
  • the cellulase(s), hemicellulase(s) and pectinase(s) represent 75% of the enzymatic mixture and the protease(s) represent 25% of said enzymatic mixture.
  • the cellulases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture
  • the hemicellulases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture
  • the pectinases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture.
  • the proteases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture.
  • the enzymatic mixture used contains 25% cellulases, 25% hemicellulases, 25% pectinases and 25% proteases.
  • the quantity used of the enzymatic mixture as defined above is between 0.25% and 10%, preferably between 1% and 6%, by volume of the water/plant material mixture.
  • the incubation according to step c) is carried out for 2 to 20 hours, preferably between 4 and 12 hours, at a temperature between 25°C and 75°C, preferably between 40°C and 60°C. , and preferably around 50°C.
  • the hydrolysis reaction is then stopped by deactivation of the enzymes by heating preferably between 80°C and 105°C for for example 5 to 20 minutes.
  • reaction medium is separated, according to step d), using all suitable known separation techniques, such as centrifugation or decantation.
  • the mixture is pre-filtered through a 0.8mm then 0.5mm sieve in order to remove the most important part of the solid phase, then sterilizing filtration is advantageously carried out on clarifying plates with a pore size of 0.2pm, at a pressure of 2 Bars.
  • the aqueous phase obtained after sterilizing filtration is stabilized with preservatives (citric acid, potassium sorbate, sodium benzoate).
  • the cosmetic composition according to the invention is characterized in that the aqueous extract of rosewood is obtained by an enzymatic extraction process comprising the steps: a) addition of water to the wood of previously crushed rose having an appropriate particle size, b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously also at least one protease, c) incubation with stirring of the previously crushed rosewood and the enzymatic mixture to release fermentable oils, proteins and sugars into the reaction medium, for a period depending on the desired yields, d) separation of the reaction medium to obtain free oil, an aqueous phase containing fermentable proteins and sugars, and a solid phase, e) separation of the aqueous phase and possible addition of preservatives.
  • an enzymatic extraction process comprising the steps: a) addition of water to the wood of previously crushed rose having an appropriate particle size, b) addition of an en
  • the aqueous extract of rosewood used according to the invention is characterized in particular by the presence of amino acids (aspartic acid, tyrosine, arginine) and total sugars (glucose, fructose).
  • the aqueous rosewood extract used according to the invention is also characterized by the fact that it is not a fermented extract.
  • the aqueous extract of rosewood used according to the invention comprises water in a content ranging from 94 to 96%, a dry extract content ranging from 1 to 6% and preferably from 3 to 5 % and preservatives in a content ranging from 0.5 to 1% by weight relative to the total weight of the extract.
  • This rosewood extract according to the invention is called INCI water (and) rose extract (and) citric acid (and) potassium sorbate (and) sodium benzoate.
  • the aqueous extract of rosewood used according to the invention will generally be present in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
  • compositions of the invention also comprise an acetyl hexapeptide and a palmitoyl tetrapeptide, in particular the following peptides:
  • Acetyl hexapeptide 8 from the company LIPOTEC marketed under the name ARGIRELINE® Amplified peptide with INCI name: WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE.
  • the commercial product ARGIRELINE® Amplified peptide containing acetyl hexapeptide-8 is present in the composition of the invention in a content ranging from 0.1% to 10% by weight relative to the total weight of the composition , preferably from 1% to 5% by weight relative to the total weight of the composition.
  • acetyl hexapeptide-8 is present in the composition of the invention in a peptide content ranging from 0.005% to 0.5% by weight relative to the total weight of the composition, preferably from 0.05% to 0.25% by weight relative to the total weight of the composition.
  • Palmitoyl tetrapeptide-7 which is a synthetic peptide with the sequence Palmitoyl-Gly-GIn-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with the INCI name: Glycerin, Lactic Acid and Palmitoyl Tetrapeptide-7.
  • the commercial product RIGIN® containing palmitoyl tetrapeptide-7 is present in the composition of the invention in a content ranging from 0.1% to 10% by weight relative to the total weight of the composition, preferably from 2% to 5% by weight relative to the total weight of the composition.
  • palmitoyl tetrapeptide-7 is present in the composition of the invention in a peptide content ranging from 0.005% to 0.5% by weight relative to the total weight of the composition, preferably from 0.1% to 0. 25% by weight relative to the total weight of the composition.
  • composition of the invention further comprises a bioactive fraction isolated from roses as described below.
  • composition of the invention will generally include:
  • bioactive fraction or “bioactive fraction isolated from roses” or “Sérum Rose De Granville®”, “Zêta” or “Zêta fraction”, we mean an extract of roses of the Evanrat variety, or rose 'Jardin de Granville®' , comprising the enzymes, proteins, sugars, ions and other active molecules present in the cytosol of the cells making up the different plant tissues of roses.
  • the extract according to the invention is distinct from a cryoextract of pink petals as described in application FR3066388.
  • the rose extract in the form of an isolated bioactive fraction used according to the invention can be prepared from fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, frozen, freeze-dried, or any mixture thereof. In the context of the invention, fresh roses are preferably used.
  • summer roses will be used to prepare the bioactive fraction isolated from roses, in particular summer rose petals.
  • winter roses in particular winter rose petals.
  • petals of roses of the Evanrat variety are rich in monosaccharide sugars (fructose, glucose, sucrose), organic acids (citric acid, malic acid), polyphenols (catechin), vitamin C, amino acids (mainly aspartic acid, glutamic acid, asparagine and glutamine), minerals (ash, potassium, calcium), and carotenoids.
  • rich in monosaccharide sugars is meant a rose extract in the form of an isolated bioactive fraction, in which said extract comprises at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose.
  • a rose extract in the form of an isolated bioactive fraction, in which said extract comprises at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
  • the composition according to the invention further comprises a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
  • a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
  • calcium improves epidermal differentiation and strengthens the structure of the skin, while potassium amplifies skin hydration and boosts energy assimilation.
  • Phyto-sugars fructtose, glucose, sucrose
  • the process implemented to obtain the isolated bioactive fraction used according to the invention comprises the main steps of: a) Cleaning of the plant material, maceration, pressing then mechanical separation of the plant material to obtain an intracellular colloidal dispersion (ICD) and a material enriched with fiber (fraction A); b) “Treatment A” then mechanical separation of the Intracellular Colloidal Dispersion (ICD) to obtain Supernatant A and a Membrane Fraction (Fraction B); c) “Treatment B” then mechanical separation of Supernatant A to obtain Supernatant B and Fraction C (Cytoplasmic Fraction); d) “Treatment C” then mechanical separation of Supernatant B to give a “Bioactive Serum Fraction” and a Fraction D (Precipitate); and e) Optionally, mixing the “bioactive serum fraction” with at least one preservative and/or stabilizer.
  • the extract of fresh roses preferably fresh rose petals, of the Evanrat variety or 'Jardin de Granville®' rose obtained at the end of this process, is a "bioactive serum fraction" or "bioactive fraction” in the sense of the invention.
  • “Cleaning” means the removal of debris from fresh Granville® roses, and preferably fresh petals, before further processing, in a manner that avoids injury to the plant, or the removal of valuable components. For example, it can be carried out by low pressure rinsing with potable water under conditions where washing with runoff water would not contain appreciably plant pigments. Excess wash water is then removed from the washed plants.
  • maceration we mean the act of transforming fresh Granville® roses, and preferably fresh petals, into smaller particles to break their integrity and subsequently facilitate the expulsion of the liquid intracellular colloidal dispersion (ICD).
  • suitable macerating instruments include, but are not limited to, devices such as a crusher, grinding stone, or grinder (e.g., knife mill, hammer mill, etc.).
  • the maceration step may include temperature monitoring and selection of maceration parameters ensuring that there is no significant increase in the temperature of the plant material during this step.
  • pressing we mean the separation of the liquid material from fresh Granville® roses, and preferably from fresh petals, by application of mechanical force. This includes, but is not limited to, techniques such as ambient gravity drainage, pressing by a heavy object, centrifugal force from a rotating expeller, piston pressure from a hydraulic press, or rollers or a screw of the appropriate type for a press.
  • fiber-enriched material or “MEF” or “FEM” (Fiber Enriched Material) we mean a solid and/or semi-solid fraction enriched with fresh Granville® rose fibers, preferably fresh petals, of which the Liquid intracellular colloid dispersion (ICD) was removed by pressing.
  • ICD Liquid intracellular colloid dispersion
  • Intracellular colloidal dispersion or “DCI” or “ICD” (Intracellular Colloidal Dispersion) is meant the liquid material expelled by pressing fresh Granville® roses, preferably fresh petals.
  • the resulting liquid contains dispersed solid and/or semi-solid particles and, potentially, water-immiscible liquid droplets of various sizes (collectively referred to as particles), in a contiguous aqueous medium.
  • the particles mainly consist of plant cell organelles, organelle fragments and fiber-enriched residual materials.
  • the aqueous medium is mainly made up of cytosols and vacuoles.
  • separation we mean the separation of solid and/or semi-solid particles, and droplets of non-aqueous liquid from an aqueous liquid by exploiting the density and/or size of the particles. This includes, but is not limited to, techniques such as dewatering, filtration (including filtration using a pressure gradient), skimming, ambient gravity sedimentation, decantation, centrifugation, or a combination of the above. Preferably, continuous flow mechanical separation will be used, but this does not exclude batch processing. “Separation 1” and “Separation 2” refer to the respective steps of the process, carried out with respective parameters.
  • supernatant is meant an aqueous material from which particles have been separated.
  • Supernatant A and Supernatant B designate the supernatants resulting from the respective separation steps of the process.
  • precipitate refers to particles from which aqueous material has been separated.
  • Fraction B and “Fraction C” designate the precipitates resulting from the respective separation steps of the process.
  • fresh Granville® rose serum fraction and “fresh Granville® rose petal serum fraction” designate compositions produced by the process as shown above without preservatives and/or stabilizers added to protect the composition of the ingredient against environmental factors such as temperature, atmosphere (e.g., oxygen), light and microorganisms.
  • preservatives and/or stabilizers we mean substances which, when added to a “fraction of fresh Granville® rose serum”, preferably a “fraction of fresh Granville® rose petal serum”, protect it against environmental factors such as temperature, atmosphere (e.g. oxygen), light and microorganisms.
  • Particular suitable substances may include, without limitation, a preservative, a stabilizer and/or a mixture thereof.
  • Granville® rose serum or “fresh Granville® rose extract” as used herein means a combination of a fraction of fresh Granville® rose serum and preservatives and/or stabilizers.
  • Gramville® rose petal serum or “Granville® fresh rose petal extract” as used herein means a combination of a fraction of Granville® fresh rose petal serum and preservatives and /or stabilizers.
  • the extract used according to the invention is obtained by implementing the extraction process disclosed in patents EP2919757, JP 6130924, CN
  • rose flowers Roshaville®
  • 4 to 5 cm of stem are harvested in such a way as to avoid chopping or crushing the collected biomass to avoid disruption of the cellular structure of the flowers.
  • Collected plants can be tested for viability using an OS5p multi-mode chlorophyll fluorometer (Opti-Sciences Inc, Hudson, NH, USA).
  • the live fresh flowers including the petals, the pistil and the stamen
  • the stem including the sepal and the receptacle, and packed in storage bags and are placed )s immediately at negative temperatures between -20°C and -80°C, such as -20°C, -40°C or -60°C.
  • the flowers can thus be stored for long periods, such as several months, or several years, before being used for the extraction process.
  • the live fresh flowers, including the petals, pistil and stamen are removed from the stem, including the sepal and receptacle, and packed into storage bags and placed in storage at a temperature between 0 and 20°C and preferably at a temperature between 2 and 6°C until the harvest is complete.
  • the roses preferably rose petals
  • the roses are immediately rinsed by spraying with water at 10°C to 15°C for 0.1 to 0.3 minutes with a flow rate of 5 to 6 liters per minute. Excess water is preferably removed from the rinsed flowers by allowing them to drain for at least 1 minute.
  • the rinsed flowers can then undergo maceration, pressing, and separation by mechanical, roller, hydraulic, or juice extractor pressing to extract the contents of the liquid intercellular colloidal dispersion (DC I) from the fiber-enriched material (“Fraction A ").
  • DC I liquid intercellular colloidal dispersion
  • the yield of fraction A is between 30% and 65%, preferably between 35% and 60% and even more preferably between 40% and 55% (weight/weight).
  • the DCI typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and even more preferably from 6% to 12% dry matter.
  • the ICD can be frozen for storage. Typically, it is frozen at -20°C.
  • the ICD When the ICD has been frozen for storage, it must first be thawed gently. Typically, it is placed to thaw at 4°C or in ice.
  • “Treatment A” is carried out by a destabilization treatment of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz.
  • the parameters of the destabilization treatment are set to obtain the reduction in the value of the real component of the low frequency dielectric constant (e'o) of approximately 20 Farads per meter (F/m) compared to its value before the treatment .
  • This processing degrades the stability of the ICD by causing agglomeration and/or aggregation of particles (i.e., organelles, organelle fragments, residual fibrous material) into assemblies large and stable enough to allow and/or or improve mechanical separation.
  • the intracellular colloidal suspension obtained as a relatively stable colloidal dispersion composed of a continuous phase (cytoplasm and vacuole contents) and a dispersed phase (suspended organelles and their fragments).
  • DLVO Derjaguin-Laundau-Verwey-Overbeek
  • DLVO theory describes the interaction and potential energy of particles based on their parameters, their distance from each other and the characteristics of the continuous phase. Changing the values of the variables affecting the repulsive force affects the stability of the dispersion. Under normal conditions of colloidal stability, an increase in potential energy as particles approach each other constitutes a potential energy barrier that cannot be overcome without external energy input. This energy barrier keeps the particles separated and the dispersion stable. The conditions modified during treatment A allow the repulsive force of the double layer to decrease to the point that there is no longer a potential energy barrier and the particles can approach and agglomerate freely.
  • Restoring the initial conditions does not restore stability, because the particles have irreversibly agglomerated. They are therefore easily removed by mechanical means (Koganov et al., sofw journal 2017).
  • the step of mechanical separation of the DCI is carried out by centrifugation in order to produce “Supernatant A” and “Fraction B”.
  • “supernatant A” has a turbidity below about 100 NTU.
  • Fraction B typically comprises from 10% to 30% dry matter, preferably from 13% to 27% dry matter, and even more preferably approximately 15.0% to 25.0% dry matter.
  • Treatment B is carried out by adjusting the pH level in the “supernatant A” by titration with, for example, an alkali until a pH greater than 6 is obtained, preferably a pH ranging from 6.5 to 7.5.
  • an alkali preferably a pH ranging from 6.5 to 7.5.
  • potassium carbonate will be used as the preferred alkali.
  • the step of mechanical separation of “Supernatant A” is preferably carried out by centrifugation in order to produce “Supernatant B” and “Fraction C”.
  • the “fraction C” typically comprises from 5% to 25% dry matter, preferably from 8% to 22% dry matter, and even more preferably approximately 10.0% to 20.0% dry matter.
  • Treatment C is carried out by adjusting the pH level in the “supernatant B”, in particular by titration with, for example, acid until a pH lower than 4.5 is obtained.
  • a solution of citric acid will be used as the preferred acid.
  • the step of mechanical separation of “Supernatant B” is preferably carried out by centrifugation in order to produce the “fresh Granville® rose serum fraction”, preferably the “serum fraction”. of fresh Granville® rose petals” (Unpreserved Extract) and “Fraction D”.
  • the “Granville® fresh rose petal serum fraction” typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and even more preferably from 6.0% to 10.0% dry matter.
  • the serum fraction obtained at the end of step d) is mixed with at least one preservative or at least one stabilizer to give a finished ingredient, or with a combination of these to give the fresh Granville® rose extract or “Granville® Rose Serum”, preferably extract of fresh Granville® rose petals or “Granville® Rose Petal Serum”.
  • Particularly suitable stabilizing agents may include, without limitation, a preservative, a stabilizer and/or mixtures thereof.
  • Suitable preservatives and stabilizers for use in the present invention include, but are not limited to, potassium sorbate, sodium benzoate, sodium metabisulfite, glycerin, propylene glycol, dipropylene glycol, butylene glycol, pentylene glycol, hexylene glycol and caprylyl glycol.
  • the stabilizing agents can comprise at least one preservative, at least one stabilizer, at least one antioxidant or mixtures thereof.
  • the bioactive fraction isolated from roses and used according to the invention is obtained by the process comprising the steps of: a) cleaning the plant material, maceration, pressing then mechanical separation of the plant material to obtain a dispersion intracellular colloid (DCI) and a fiber-enriched material (fraction A); b) destabilization of the DCI with electromagnetic waves then mechanical separation of the intracellular colloidal dispersion (DCI) to obtain supernatant A and a Membrane Fraction (fraction B); c) adjustment of the pH level in supernatant A until obtaining a pH greater than 6, preferably a pH ranging from 6 to 7, then mechanical separation of supernatant A to obtain supernatant B and fraction C (cytoplasmic fraction) ; d) adjustment of the pH level in the supernatant B until obtaining a pH lower than 4.5 then mechanical separation of the supernatant B to give a “Bioactive serum fraction” and a fraction D (precipitate); and e) Optionally, mixing the “bioactive serum fraction” and a fraction
  • the bioactive fraction isolated from roses, preferably from rose petals, used according to the invention comprises water in a content ranging from 90 to 94%, a dry extract content ranging from 6 to 10% .
  • This bioactive fraction isolated from roses used according to the invention is called INCI name Rosa Hybrid Flower Extract.
  • the bioactive fraction isolated from roses, preferably from rose petals, obtained according to the invention comprises water in a content ranging from 89 to 93%, a dry extract content ranging from 6 to 10 % and preservatives and/or stabilizers in a content ranging from 0.3 to 1% by weight relative to the total weight of the extract.
  • the bioactive fraction isolated from roses may be present in the composition of the invention in a content ranging from 0.1 to 1% by weight, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
  • the cosmetic composition according to the invention comprises: 1. an aqueous extract of rosewood of the Evanrat variety in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition,
  • hexapeptide with the INCI name Acetyl Hexapeptide-8 in a content ranging from 0.05% to 0.25% by weight (of peptide) relative to the total weight of the composition,
  • a tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7 in a content ranging from 0.1% to 0.25% by weight (of peptide) relative to to the total weight of the composition, and
  • a bioactive fraction isolated from fresh roses of the Evanrat variety in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
  • cosmetic composition any composition for cosmetic purposes, that is to say aesthetic, which can be brought into contact with the superficial parts of the human body and more particularly with keratin materials, in particular the skin and/or the lips, particularly the skin of the face and/or neck.
  • physiologically acceptable medium any excipient suitable for topical use, in contact with keratin materials, without risk of toxicity, incompatibility, instability and/or allergic response.
  • the physiologically acceptable medium generally represents 1 to 99% by weight, relative to the total weight of said composition.
  • topical use or “topical application” is meant a composition intended for application to keratin materials. In fact, an oral composition is not intended for topical application to the skin.
  • the cosmetic composition according to the invention is characterized in that it is a care and/or makeup composition for the skin and/or lips, in particular a composition skin care.
  • a composition for topical application according to the invention may for example be in the form of a cream, oil-in-water emulsion, or water-in-oil or multiple emulsion, solution, suspension, gel, milk, lotion, serum , balm, stick, or even powder.
  • the cosmetic composition of the invention is in the form of a cream, oil-in-water emulsion, or water-in-oil or multiple emulsion, solution, suspension, gel, milk, lotion, or serum .
  • said composition used according to the invention is in the form of a cream or a serum.
  • the aqueous phase of the composition according to the invention comprises water and optionally a water-soluble solvent.
  • water-soluble solvent' is meant a compound that is liquid at room temperature and miscible with water (miscibility in water greater than 50% by weight at 25°C and atmospheric pressure).
  • lower C1 -C5 mono-alcohols such as ethanol, isopropanol and their mixtures
  • C2-C8 glycols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, dipropylene glycol, and mixtures thereof
  • C2-C32 polyols such as polyglycerols, polyethylene glycols, and mixtures thereof, and mixtures thereof.
  • hydrophilic gelling agents may also include hydrophilic gelling agents, antioxidants, preservatives and mixtures thereof.
  • the cosmetic composition according to the invention may also comprise a fatty phase (solid fatty substances) or oily phase.
  • oil phase is meant an oil or a mixture of oils which may or may not be miscible with each other.
  • oil is meant, within the meaning of the invention, a fatty substance, not soluble in water, liquid at 25°C and atmospheric pressure. These oils can be volatile or non-volatile, vegetable, mineral or synthetic.
  • An oily phase according to the invention may comprise natural, hydrocarbon, silicone oils, and mixtures thereof.
  • the content of fatty or oily phase in the cosmetic composition of the invention will generally range from 0.2% to 45%, preferably from 0.5% to 30%, and more preferably from 2% to 25% by weight per relative to the total weight of said composition.
  • composition of the invention may also comprise any additive usually used in cosmetics such as antioxidants, perfumes, cosmetic active agents, such as for example emollient agents, moisturizing agents, vitamins, anti-aging agents, lifting agents, tightening agents, plumping agents, lightening agents, fillers, pearls and their mixtures.
  • cosmetic active agents such as for example emollient agents, moisturizing agents, vitamins, anti-aging agents, lifting agents, tightening agents, plumping agents, lightening agents, fillers, pearls and their mixtures.
  • the invention relates to a cosmetic composition for topical application to the skin and/or lips, in particular the skin of the face and/or body, comprising in a physiologically acceptable medium, at least one aqueous extract of rosewood, Acetyl Hexapeptide-8, Palmitoyl tetrapeptide-7 and optionally an isolated bioactive fraction obtained from fresh rose, and at least one cosmetic adjuvant chosen from the group consisting of antioxidants, perfumes , emollient agents, moisturizing agents, vitamins, anti-aging agents, lifting agents, tightening agents, plumping agents, lightening agents, fillers, pearls and mixtures thereof.
  • the invention also relates to a non-therapeutic cosmetic process for caring for and/or making up the skin and/or lips comprising the application to the skin and/or lips, in particular the skin, of a composition such as defined according to the invention.
  • skin and/or lips we mean in particular healthy skin and/or lips, that is to say not presenting disorders or disorders which would relate to a pathological state (subjects ' unhealthy', suffering from a pathology).
  • the application of the composition as defined according to the invention to the skin and/or lips makes it possible to prevent and/or reduce the signs of skin aging, in particular the loss of density and/or firmness. of the skin, the reduction in the thickness of the epidermis, the reduction in the differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines. Association of assets and uses
  • the present invention also relates to a combination of cosmetic active ingredients comprising:
  • an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
  • the invention also relates to the non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined above, as an active association for preventing and/or reducing the signs of skin aging, in particular loss of density and/or or firmness of the skin, the reduction in the thickness of the epidermis, the reduction in the differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines.
  • the present invention also relates to a cosmetic set comprising:
  • an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium, characterized in that at least one of the active ingredients ( i) to (iv) is packaged in a composition distinct from the other active ingredients, and in particular each active ingredient is packaged in a distinct composition.
  • Another object is a cosmetic process comprising the successive application, to the skin and/or lips, of the compositions of the cosmetic set as defined above.
  • Fibroblasts from elderly donors were trypsinized and counted.
  • the cells were suspended in the bioink consisting of alginate, gelatin and fibrinogen (LabSkin Creations) at a rate of 250,000 cells per mL of ink.
  • the bioinks were transferred into sterile syringes mounted on 10 gauge cannulas.
  • the printed objects were modeled with the software Slic3r (GNU Affero General Public License) and Repetier (Hot-World GmbH & Co. KG).
  • the dermis were printed using a 3D printer (LabSkin Creations) to a size of 1.5cm x 1.5cm and had two layers of cells extruded into the bioink.
  • the constructs were then polymerized for 1 hour in a solution of calcium and thrombin (LabSkin Creations) allowing the consolidation of the bioink. After three rinses in a saline buffer, the equivalent dermis were cultured in a container of DMEM supplemented with 10% calf serum and 1% antibiotics and antifungal and placed in the incubator (37°C, 5 % CO2) for 12 days. The culture medium was renewed every 2 days.
  • Confluent keratinocytes were trypsinized and then counted. The equivalent of 500,000 cells was seeded on the surface of each bioprinted dermis. After 30 minutes of adhesion at 37°C, culture medium containing DMEM supplemented with 10% fetal bovine serum, hydrocortisone and insulin was added to submerge the bioprinted dermo-epidermal constructs.
  • bioprinted dermo-epidermal assemblies were cultured for 7 days in culture medium containing DMEM supplemented with 10% fetal calf serum, hydrocortisone and insulin allowing the proliferation of keratinocytes. They were then raised at the air/liquid interface in order to initiate the differentiation of the epidermis. This final 7-day culture step made it possible to obtain multistratified epidermis.
  • Treatments with active ingredients alone or in combinations were dissolved in the culture media.
  • the final concentrations of active ingredients are specified for each example.
  • Step 1 passage through a garden shredder to obtain sections of two centimeters
  • Step 2 passage through an industrial shredder
  • the enzymatic extraction was carried out in a reactor with a maximum capacity of 70 I.
  • the reactor is stirred by propeller and is thermostatically controlled by a double jacket.
  • the mixture was raised to a temperature of 85°C for 1 min and then stabilized at 50°C.
  • HEL1 PR1 Enzymatic cocktail comprising a mixture of cellulases, hemicellulases and pectinases at 75% and proteases at 25%, were added to the corresponding reaction mixture (5% w/w). The reaction is continued for 4 hours, the temperature is maintained at 50°C.
  • the mixture was heated to 85°C for 1 min to inactivate the enzymes, then returned to 35°C.
  • the mixture is pre-filtered through a 0.8mm then 0.5mm sieve in order to remove the most important part of the solid phase, then on a sheet. Sterilizing filtration
  • the aqueous phase obtained after sterilizing filtration is stabilized with the following preservatives:
  • an aqueous extract of rosewood is obtained with 3.7% by weight of dry matter relative to the total weight of the extract.
  • the bioactive fraction of rose petals used according to the invention is obtained according to the following protocol: a1) cleaning the fresh rose petals by spraying with water at a temperature of approximately 12°C, for 0.1 to 0.3 minutes at a flow rate of 5 to 6 liters per minute, a2) maceration, pressing then mechanical separation of the roses using a mechanical screw press (model CP-6 Vincent Corporation, FL) to extract the contents the liquid intercellular colloidal dispersion (DCI) of the fiber-enriched material (“Fraction A”); b1) destabilization of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz, b2) centrifugation of the intracellular colloidal dispersion (DCI) to obtain the supernatant A and a Membrane Fraction ( fraction B); c1) adjustment of the pH level in the supernatant A using an alkali (Le., potassium carbonate), in order to obtain a pH ranging from 6 to 7, c2) mechanical separation of the supernatant A to obtain the super
  • Granville® rose petal serum also called “Zeta serum”, or “Zf serum” or “ZETA FRACTION” or “ZETA fraction” in the following examples
  • Zf serum also called “Zf serum” or “ZETA FRACTION” or “ZETA fraction” in the following examples
  • ZETA fraction contains 8% dry matter (active ingredient), 91.5 % by weight water, 0.15% by weight potassium sorbate and 0.3% by weight sodium benzoate. These contents are by weight relative to the total weight of the extract.
  • the INCI name for this extract is Rosa Hybrid Flower Extract, Potassium Sorbate, and Sodium Benzoate.
  • Acetyl hexapeptide 8 from the company LIPOTEC is marketed under the name ARGIRELINE® Amplified peptide with INCI name: WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE.
  • Acetyl hexapeptide 8 means the commercial product ARGIRELINE® Amplified peptide comprising 0.05% peptide.
  • the percentages indicated in the examples correspond to the percentages by weight of commercial product in culture media or in cosmetic formulas.
  • Palmitoyl tetrapeptide-7 is a synthetic peptide with the sequence Palmitoyl-Gly-GIn-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with the INCI name: Glycerin, Lactic Acis and Palmitoyl Tetrapeptide-7.
  • “Palmitoyl tetrapeptide-7” means the commercial product RIGIN® comprising 0.05% peptide. The percentages indicated in the examples correspond to the percentages by weight of commercial product in culture media or in cosmetic formulas.
  • the concentrations of active ingredients or combinations were selected based on the pre-performed cytotoxicity results.
  • the reconstructed skins were treated during dermal reconstruction, from D5 to D 12 and epidermal from D14 to D18 then from D20 to D26.
  • Paraffin sections of 5 ⁇ m were made on the different skin samples reconstructed by bioprinting. After deparaffinization and rehydration, the samples were stained with Hematoxylin, Phloxine and Saffron (HPS) in three successive 5 min baths. After rinsing, the sections were dehydrated before being mounted on coverslips with a hydrophobic xylene-based mounting medium.
  • HPS Hematoxylin, Phloxine and Saffron
  • Image analysis was carried out with Imaged software (Rasband, WS, Imaged, US National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997-2017). For each parameter a statistical analysis was carried out with a Student test, which is significant if its value is less than 0.05. The results are indicated according to: * P ⁇ 0.05, ** P ⁇ 0.01 and *** P ⁇ 0.001.
  • the thickness of the cohesive layers of the epidermis is measured using a Euclidean distance map.
  • the pixels corresponding to the epidermis are segmented from the other pixels of the image.
  • the segmented image is then binarized and converted into a Euclidean distance map.
  • the basement membrane of the epidermis is selected and applied to the distance map to measure at each point the distance between the basement membrane and the terminal layers of the epidermis.
  • the value obtained corresponds to the average distance between the basement membrane and the terminal layers of the epidermis. 9 images are analyzed for each condition.
  • Example 2 Effects of the active ingredients and combinations according to the invention on the KI67 factor The effect of the active ingredients alone or in combinations according to the invention on the production of the KI67 factor by the skin samples reconstructed by bioprinting was analyzed. Each test was performed in triplicate.
  • the skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations (dissolved in the culture media): the aqueous extract of rosewood alone (final concentration 0.07%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%) + aqueous extract of rosewood (final concentration 0.025%).
  • Example 3 Effects of active ingredients and combinations according to the invention on the thickness of the dermis
  • the skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations (dissolved in the culture media): the aqueous extract of rosewood alone (final concentration 0.07%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%) + aqueous extract of rosewood (final concentration 0.025%).
  • the skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations according to the invention (dissolved in the culture media): the bioactive fraction isolated from fresh roses, otherwise called ZETA fraction, alone (final concentration 0.05% ) ; a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.21 g/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.06%) + aqueous extract of rosewood (final concentration 0.005%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.10pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.003%) + aqueous extract of rosewood (final concentration 0.004%) + ZETA fraction ( final concentration 0.003%).
  • Example 5 Cosmetic formulations according to the invention 5.1 Composition in the form of an emulsion
  • Carbomer (Carbopol® 981) 0.3%
  • Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)
  • Palmitoyl tetrapeptide-7 (RIGIN® comprising
  • this composition gives tone, elasticity and firmness, the features are enhanced, the face is reshaped.
  • Carbomer (Carbopol® 981) 0.3%
  • Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 2% (i.e. 0.001% peptide) Palmitoyl tetrapeptide-7 (RIGIN® comprising
  • this composition gives tone, elasticity and firmness, the features are enhanced, the face is reshaped.
  • Lecithin and sodium acrylates copolymer (Lecigel PCR negative) 1.20%
  • Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)
  • Palmitoyl tetrapeptide-7 (RIGIN® comprising 0.05% peptide) 4% (i.e. 0.002% peptide)

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Abstract

The present invention relates to a cosmetic composition comprising, in a physiologically acceptable medium: (i) an aqueous extract of rose wood of the Evanrat variety, preferably of summer rose wood, obtained by enzymatic extraction by means of a cosmetically acceptable polar solvent, (ii) a hexapeptide with the INCI name: Acetyl Hexapeptide-8, (iii) a tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, and (iv) optionally also an extract of roses in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety.

Description

Composition anti-âge avec extraits de rose et peptides Anti-aging composition with rose extracts and peptides
DOMAINE DE L'INVENTION FIELD OF THE INVENTION
La présente invention concerne une composition cosmétique, en particulier une composition cosmétique de soin destinée à prévenir et/ou diminuer les signes du vieillissement cutané, en particulier de la peau du visage et/ou du corps. The present invention relates to a cosmetic composition, in particular a cosmetic care composition intended to prevent and/or reduce the signs of skin aging, in particular of the skin of the face and/or body.
ETAT DE LA TECHNIQUE STATE OF THE ART
La peau est la première barrière de protection de l'organisme vis-à-vis de l'environnement. Elle est quotidiennement soumise aux effets de facteurs d’origine exogène (ex : rayonnements UV, variations de température, pollution atmosphérique, fumée de cigarette...) et/ou endogène (ex : hormones...). The skin is the body's first protective barrier against the environment. It is daily subject to the effects of factors of exogenous origin (e.g.: UV radiation, temperature variations, atmospheric pollution, cigarette smoke, etc.) and/or endogenous (e.g.: hormones, etc.).
Le vieillissement entraîne des altérations au niveau de la peau du corps et/ou du visage y compris des lèvres, zones particulièrement soumises aux effets délétères de l’environnement, telles que des rides et ridules, une peau ayant perdu sa fermeté, sa souplesse et son élasticité, une diminution de l’épaisseur de l’épiderme liée notamment à un ralentissement du renouvellement des cellules de la peau. Aging leads to changes in the skin of the body and/or face, including the lips, areas particularly subject to the deleterious effects of the environment, such as wrinkles and fine lines, skin having lost its firmness, suppleness and its elasticity, a reduction in the thickness of the epidermis linked in particular to a slowdown in the renewal of skin cells.
On connaît de l’art antérieur de très nombreux actifs anti-âge tels que des peptides, des vitamines, des antioxydants et des extraits végétaux. On peut citer notamment les peptides Acetyl Hexapeptide-8 de la société Lipotec et Palmitoyl Tetrapeptide-7 de la société Sederma. A large number of anti-aging active ingredients such as peptides, vitamins, antioxidants and plant extracts are known from the prior art. Mention may in particular be made of the peptides Acetyl Hexapeptide-8 from the company Lipotec and Palmitoyl Tetrapeptide-7 from the company Sederma.
On connaît également l’utilisation de certains extraits de roses en anti-âge comme l’extrait aqueux de bois de rose de la variété Evanrat décrit dans la demande de brevet FR3090380. We also know the use of certain rose extracts for anti-aging, such as the aqueous extract of rosewood of the Evanrat variety described in patent application FR3090380.
Toutefois, il existe un besoin constant de trouver de nouveaux composés ou de nouvelles associations d’actifs ayant des propriétés améliorées dans la prévention et/ou le vieillissement de la peau. However, there is a constant need to find new compounds or new combinations of active ingredients with improved properties in the prevention and/or aging of the skin.
De manière inattendue, la Demanderesse a mis en évidence un effet in vitro anti-âge synergique entre les peptides (Acetyl Hexapeptide-8 et le Palmitoyl tétrapeptide-7) et un extrait aqueux de bois de rose de la variété Evanrat. Elle a également démontré une synergie anti-âge entre les peptides (Acetyl Hexapeptide-8 et le Palmitoyl tétrapeptide-7), un extrait aqueux de bois de rose de la variété Evanrat et un extrait de roses sous la forme d’une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat. Unexpectedly, the Applicant has demonstrated a synergistic anti-aging in vitro effect between the peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7) and an aqueous extract of rosewood of the Evanrat variety. It also demonstrated anti-aging synergy between peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7), an extract aqueous rosewood of the Evanrat variety and a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety.
EXPOSE DE L'INVENTION STATEMENT OF THE INVENTION
La présente invention porte donc notamment sur une composition cosmétique comprenant, dans un milieu physiologiquement acceptable : The present invention therefore relates in particular to a cosmetic composition comprising, in a physiologically acceptable medium:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, et (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8, and
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7. (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7.
La présente invention porte encore sur un procédé cosmétique non thérapeutique de soin et/ou de maquillage de la peau et/ou des lèvres comprenant l’application sur ladite peau et/ou lesdites lèvres, en particulier la peau, d’une composition cosmétique telle que définie dans la présente invention. The present invention also relates to a non-therapeutic cosmetic process for caring for and/or making up the skin and/or lips comprising the application to said skin and/or said lips, in particular the skin, of a cosmetic composition such as as defined in the present invention.
L’invention porte également sur une association d’actifs cosmétiques comprenant : The invention also relates to a combination of cosmetic active ingredients comprising:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8,
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7, et (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, and
(iv) Eventuellement en outre une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. (iv) Optionally also an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
L’invention porte en outre sur l’utilisation cosmétique non thérapeutique de l’association d’actifs cosmétiques telle que définie dans la présente invention, comme association active pour prévenir et/ou diminuer les signes du vieillissement cutané, en particulier la perte de densité et/ou de fermeté de la peau, la diminution de l’épaisseur de l’épiderme, la diminution de la différenciation et/ou prolifération des cellules épidermiques, et/ou l’apparition de rides et/ou ridules. The invention further relates to the non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined in the present invention, as an active association for preventing and/or reducing the signs of skin aging, in particular loss of density. and/or firmness of the skin, reduction in the thickness of the epidermis, reduction differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines.
L’invention porte encore sur un ensemble cosmétique comprenant : The invention also relates to a cosmetic set comprising:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8,
(iii) Un tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7, et (iii) A tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and
(iv) Eventuellement en outre une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium, caractérisé en ce que l’un au moins des actifs (i) à (iv) est conditionné dans une composition distincte des autres actifs, et en particulier chaque actif est conditionné dans une composition distincte. (iv) Optionally also an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium, characterized in that at least one of the active ingredients (i) to (iv) is packaged in a composition distinct from the other active ingredients, and in particular each active ingredient is packaged in a distinct composition.
L’invention porte enfin sur un procédé cosmétique comprenant l’application successive, sur la peau et/ou les lèvres, des compositions de l’ensemble cosmétique tel que défini dans la présente invention. The invention finally relates to a cosmetic process comprising the successive application, to the skin and/or lips, of the compositions of the cosmetic assembly as defined in the present invention.
DESCRIPTION DES FIGURES DESCRIPTION OF FIGURES
[Fig. 1] Effet des différents actifs et associations d’actifs selon l’invention sur l’expression protéique du facteur KI67 dans des cultures de peau humaine reconstruite. [Fig. 1] Effect of the different active ingredients and combinations of active ingredients according to the invention on the protein expression of the KI67 factor in cultures of reconstructed human skin.
[Fig. 2] Effet des différents actifs et associations d’actifs selon l’invention sur l’épaisseur de l’épiderme dans des cultures de peau humaine reconstruite. [Fig. 2] Effect of the different active ingredients and combinations of active ingredients according to the invention on the thickness of the epidermis in cultures of reconstructed human skin.
[Fig. 3] Effet des différents actifs et associations d’actifs selon l’invention sur l’expression protéique de Collagène VII dans des cultures de peau humaine reconstruite. [Fig. 3] Effect of the different active ingredients and combinations of active ingredients according to the invention on the protein expression of Collagen VII in cultures of reconstructed human skin.
DESCRIPTION DETAILLEE DE L'INVENTION DETAILED DESCRIPTION OF THE INVENTION
Ainsi, la présente invention porte donc notamment sur une composition cosmétique comprenant, dans un milieu physiologiquement acceptable : (i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, Thus, the present invention therefore relates in particular to a cosmetic composition comprising, in a physiologically acceptable medium: (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, et (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8, and
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7. (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7.
Les roses ou rosiers utilisés comme matériel végétal pour la préparation des extraits utilisés selon l’invention correspondent au rosier ou rose ‘Jardin de Granville®’, une variété hybride proposée exclusivement par « Roses anciennes André Eve S.A.S » et protégée par Certificat d’Obtention Végétale sous le N°20110345 avec pour espèce la dénomination Rosa L. et pour la variété la dénomination EVANRAT. Ce rosier buisson appartient au groupe des hybrides modernes, qui, de mai à octobre, se couvre de roses en permanence, montrant ainsi un excellent caractère remontant. The roses or rose bushes used as plant material for the preparation of the extracts used according to the invention correspond to the rose bush or rose 'Jardin de Granville®', a hybrid variety offered exclusively by "Roses ancien André Eve S.A.S" and protected by Certificate of Obtention Plant under number 20110345 with the species name Rosa L. and the variety the name EVANRAT. This bush rose belongs to the group of modern hybrids, which, from May to October, is permanently covered with roses, thus showing an excellent repeating character.
Extrait de bois de rose et procédé de préparation Rosewood extract and preparation process
L’extrait aqueux de bois de rose de la variété Evanrat ou rose ‘Jardin de Granville®’, de préférence de bois de rose d’été, utilisé selon l’invention est obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, selon les techniques décrites dans la demande FR3090380. The aqueous extract of rosewood of the Evanrat variety or 'Jardin de Granville®' rose, preferably summer rosewood, used according to the invention is obtained by enzymatic extraction using a cosmetically acceptable polar solvent. , according to the techniques described in application FR3090380.
On parlera indifféremment d’extrait aqueux de bois de rose ou de bois de rosier ou d’extrait polaire de bois de rose ou de bois de rosier ou d’extrait hydrophile de bois de rose ou de bois de rosier ou d’éco-extrait obtenu par un procédé d’éco-extraction de bois de rose ou de bois de rosier dans la description. We will speak indifferently of aqueous extract of rosewood or rosewood or polar extract of rosewood or rosewood or hydrophilic extract of rosewood or rosewood or eco-extract obtained by an eco-extraction process of rosewood or rosewood in the description.
L’expression « solvant polaire » signifie que le solvant a une valeur d’indice de Polarité qui est égale ou supérieure à une valeur de 4. L’indice de polarité est une grandeur calculée sur la base de grandeurs thermodynamiques (de solubilité et de changement d’état) qui met en évidence le caractère plus ou moins polaire d’une molécule. On se reportera, pour les indices de polarité des solvants, à l'article de L.R. SNYDER : Classification of the solvent properties of common liquids ; Journal of Chromatography, 92 (1974), 223-230. The expression "polar solvent" means that the solvent has a Polarity index value which is equal to or greater than a value of 4. The polarity index is a quantity calculated on the basis of thermodynamic quantities (solubility and change of state) which highlights the more or less polar nature of a molecule. Refer, for the polarity indices of solvents, to the article by L.R. SNYDER: Classification of the solvent properties of common liquids; Journal of Chromatography, 92 (1974), 223-230.
Les solvants polaires préférés sont ceux constitués d’un composé comprenant au moins une liaison covalente polaire de type O-H. A titre de solvant polaire cosmétiquement acceptable, on choisit un solvant ou un mélange de solvants choisi(s) parmi l’eau, les alcools en C1-C4, tel que l’éthanol, les glycols, tels que l’éthylène glycol, le glycérol, le butylèneglycol et le propylèneglycol, et leurs mélanges. De préférence, on utilise l’eau. The preferred polar solvents are those consisting of a compound comprising at least one polar covalent bond of the OH type. As a cosmetically acceptable polar solvent, a solvent or a mixture of solvents chosen from water, C1-C4 alcohols, such as ethanol, glycols, such as ethylene glycol, glycerol, butylene glycol and propylene glycol, and mixtures thereof. Preferably, water is used.
Selon un mode particulier, ledit extrait est caractérisé en ce qu’il comprend de l’eau en une teneur allant de 94 à 96%, une teneur en extrait sec allant de 3 à 5% et des conservateurs en une teneur allant de 0,5 à 1 % en poids par rapport au poids total de l’extrait. According to a particular mode, said extract is characterized in that it comprises water in a content ranging from 94 to 96%, a dry extract content ranging from 3 to 5% and preservatives in a content ranging from 0, 5 to 1% by weight relative to the total weight of the extract.
Matériel végétal Plant material
On distingue deux types de bois de rose selon le moment de l’année où ils sont récoltés :There are two types of rosewood depending on the time of year they are harvested:
- Les bois d’hiver, généralement récoltés entre les mois de novembre et d’avril - Winter wood, generally harvested between the months of November and April
- Les bois d’été, généralement récoltés entre les mois de mai et d’octobre. - Summer wood, generally harvested between the months of May and October.
Selon un mode particulier et préféré, on utilisera des bois d’été. On parlera indifféremment dans la description de bois de rose ou bois de rose d’été. According to a particular and preferred method, we will use summer wood. We will speak indifferently in the description of rosewood or summer rosewood.
Les bois de rose selon l’invention sont obtenus à partir de roses entières fraichement coupées et sont isolés du reste de la plante par séparation manuelle ou mécanique. The rosewood according to the invention is obtained from freshly cut whole roses and is isolated from the rest of the plant by manual or mechanical separation.
Dans un mode de réalisation particulier, les bois de rose sont directement utilisés en vue de l’extraction. In a particular embodiment, the rosewoods are directly used for extraction.
Dans un autre mode de réalisation, les bois de rose sont préalablement séchés avant extraction. In another embodiment, the rosewood is previously dried before extraction.
Selon un mode particulier et préféré, le bois de rose est broyé préalablement à l’étape d’extraction. According to a particular and preferred method, the rosewood is crushed before the extraction stage.
Les bois de rose comprennent des sucres monosaccharides (fructose, glucose), et polysaccharides en particulier disaccharides, des polyphénols (catéchine), des acides aminés (majoritairement acide aspartique tyrosine et arginine), des minéraux (cendres, sodium, potassium, calcium), des flavonoïdes (pour la plupart glycosylés) et des tanins. Rosewood includes monosaccharide sugars (fructose, glucose), and polysaccharides in particular disaccharides, polyphenols (catechin), amino acids (mainly aspartic acid, tyrosine and arginine), minerals (ash, sodium, potassium, calcium), flavonoids (mostly glycosylated) and tannins.
En particulier, le calcium améliore la différenciation épidermique et renforce la structure de la peau, tandis que le potassium amplifie l’hydratation cutanée et booste l’assimilation de l’énergie. Les phytosucres (fructose, glucose) sont destinés à gorger les cellules cutanées d’énergie. L’extrait aqueux de bois de rose selon l’invention est un extrait concentré en composés naturels polaires. L’extrait aqueux de bois de rose selon l’invention est distinct d’une eau de rose. Il s’agit ici d’un extrait de bois de rose mettant en oeuvre l’extraction de composés naturels de bois de rose en présence de solvants aqueux (polaire) avec ou sans ajout de modificateur de pH, et une mise en oeuvre, dans la formulation, sous la forme d’une solution ou concentré aqueux, le solvant du concentré pouvant être le solvant d’extraction et/ou un solvant additionnel. In particular, calcium improves epidermal differentiation and strengthens the structure of the skin, while potassium amplifies skin hydration and boosts energy assimilation. Phytosugars (fructose, glucose) are intended to fill skin cells with energy. The aqueous rosewood extract according to the invention is an extract concentrated in polar natural compounds. The aqueous extract of rosewood according to the invention is distinct from rose water. This is a rosewood extract using the extraction of natural rosewood compounds in the presence of aqueous solvents (polar) with or without the addition of a pH modifier, and an implementation, in the formulation, in the form of an aqueous solution or concentrate, the solvent of the concentrate possibly being the extraction solvent and/or an additional solvent.
Selon un mode particulier, l’extrait aqueux de bois de rose selon l’invention comprend en outre en faible proportion un extrait de fleurs de rose représentant de 0,01 à 1 %, de préférence de 0,05 à 0,5%, de préférence 0,1 à 0,2% de l’extrait aqueux total de rose selon l’invention. Dans ce mode de réalisation, le matériel végétal d’origine comprend des bois de rose et des fleurs de rose dans une proportion 0,01 % fleurs pour 99,99% de bois à 1 % fleurs pour 99% de bois. According to a particular embodiment, the aqueous extract of rosewood according to the invention further comprises in a small proportion an extract of rose flowers representing from 0.01 to 1%, preferably from 0.05 to 0.5%, preferably 0.1 to 0.2% of the total aqueous rose extract according to the invention. In this embodiment, the original plant material comprises rosewood and rose flowers in a proportion 0.01% flowers for 99.99% wood to 1% flowers for 99% wood.
Procédé d’extraction Extraction process
L’extrait aqueux de bois de rose selon l’invention peut être obtenu selon différents procédés d’éco-extraction connus de l’homme du métier et notamment celui décrit ci-après. The aqueous extract of rosewood according to the invention can be obtained according to different eco-extraction processes known to those skilled in the art and in particular that described below.
Selon un mode particulier et préféré, on utilisera comme matériel végétal dans les procédés d’extraction adaptés à l’invention décrits ci-après, des bois de rose de la variété Evanrat, de préférence la rose Jardin de Granville®, de préférence des bois de rose préalablement séchés. According to a particular and preferred mode, we will use as plant material in the extraction processes adapted to the invention described below, rosewood of the Evanrat variety, preferably the Jardin de Granville® rose, preferably wood of roses previously dried.
Les bois peuvent être utilisés frais, congelés ou préalablement séchés à l’aide d’outils de séchage conventionnels et connus de l’homme du métier, comme par exemple le séchage à l’air libre, en étuves, par lyophilisation ou zéodratation. Ainsi, selon un mode de réalisation, et préalablement à l’étape d’extraction elle-même, les bois peuvent avoir été séchés et/ou broyés. Selon un autre mode de réalisation, les bois sont utilisés humides puis broyés. The wood can be used fresh, frozen or previously dried using conventional drying tools known to those skilled in the art, such as drying in the open air, in ovens, by freeze-drying or zeodration. Thus, according to one embodiment, and prior to the extraction step itself, the wood may have been dried and/or crushed. According to another embodiment, the wood is used wet then crushed.
Selon un mode particulier, les bois sont broyés avant extraction, par exemple au moyen d’un mortier, d’un cryobroyage, d’un mixeur, d’un broyeur traditionnel ou d’un centrifugeur selon les méthodes connues de l’homme du métier. According to a particular method, the wood is crushed before extraction, for example by means of a mortar, cryo-grinding, a mixer, a traditional grinder or a centrifuge according to methods known to those skilled in the art. job.
L’extrait aqueux de bois de rose selon l’invention peut être obtenu en particulier selon des procédés d’extraction enzymatique. Selon les molécules que l’on cherche à extraire, les enzymes seront adaptées aux parois à hydrolyser pour accéder à ces dites molécules. Avantageusement, on pourra notamment utiliser des enzymes de type cellulase, hémicellulase, pectinase ou protéase, seules ou en combinaison. The aqueous extract of rosewood according to the invention can be obtained in particular according to enzymatic extraction processes. Depending on the molecules that we seek to extract, the enzymes will be adapted to the walls to be hydrolyzed to access these said molecules. Advantageously, it is possible in particular to use enzymes of the cellulase, hemicellulase, pectinase or protease type, alone or in combination.
On pourra ainsi utiliser une cellulase pour libérer le glucose et la cellulose ; une hemicellulase pour libérer les mono et oligomères de sucres réduits ; une pectinase pour libérer les acides uroniques ; et avantageusement une protéase pour hydrolyser les protéines de structure et améliorer la lyse de la matière première. We can thus use a cellulase to release glucose and cellulose; a hemicellulase to release reduced sugar mono- and oligomers; a pectinase to release uronic acids; and advantageously a protease to hydrolyze the structural proteins and improve the lysis of the raw material.
Dans un mode de réalisation préféré, l’éco-extraction est réalisée au moyen du protocole décrit dans la demande de brevet WO2011045387 (correspondant à la demande de brevet FR2351461) au nom de l’institut National Polytechnique de Lorraine INPL. Il s’agit d’une technologie alternative répondant aux contraintes réglementaires et environnementales tout en restant compétitif économiquement. Ce procédé innovant, permet d’extraire de la matière végétale, en une seule étape, trois types de produits à savoir une huile (riche en nutriments polyphénols, stérols, vitamine E...), un extrait aqueux ainsi qu’un tourteau (phase solide).In a preferred embodiment, eco-extraction is carried out using the protocol described in patent application WO2011045387 (corresponding to patent application FR2351461) in the name of the National Polytechnic Institute of Lorraine INPL. This is an alternative technology that meets regulatory and environmental constraints while remaining economically competitive. This innovative process makes it possible to extract from plant material, in a single step, three types of products, namely an oil (rich in polyphenol nutrients, sterols, vitamin E, etc.), an aqueous extract as well as a cake ( solid phase).
Ainsi, dans un mode de réalisation de l’invention, l’extrait aqueux de bois de rose utilisé selon l’invention est obtenu par un procédé d’extraction enzymatique comprenant les étapes comprenant les étapes : a) addition d'eau au bois de rose préalablement broyé présentant une taille de particule appropriée, b) addition d'un mélange enzymatique contenant au moins une cellulase, au moins une hémicellulase et au moins une pectinase, et avantageusement en outre au moins une protéase, c) incubation sous agitation du bois de rose préalablement broyé et du mélange enzymatique pour libérer dans le milieu réactionnel des huiles, des protéines et des sucres fermentescibles, pendant une durée dépendant des rendements recherchés, d) séparation du milieu réactionnel pour obtenir de l'huile libre, une phase aqueuse contenant des protéines et des sucres fermentescibles, et une phase solide, et e) séparation de la phase aqueuse et ajout éventuel de conservateurs. Thus, in one embodiment of the invention, the aqueous extract of rosewood used according to the invention is obtained by an enzymatic extraction process comprising the steps comprising the steps: a) addition of water to the wood of previously crushed rose having an appropriate particle size, b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously also at least one protease, c) incubation with stirring of the wood of previously crushed rose and the enzymatic mixture to release into the reaction medium oils, proteins and fermentable sugars, for a period depending on the desired yields, d) separation of the reaction medium to obtain free oil, an aqueous phase containing fermentable proteins and sugars, and a solid phase, and e) separation of the aqueous phase and possible addition of preservatives.
Ledit extrait de bois de rose utilisé selon l’invention étant obtenu après l’étape de séparation (e). Selon un mode particulier, à l’étape a), la taille de particule appropriée de la matière végétale est avantageusement obtenue par broyage de ladite matière végétale. Le broyage doit être le plus fin possible pour favoriser l'action des enzymes. Idéalement, toutes les particules doivent avoir une taille proche de 50 pm, de préférence proche de 10 pm. Said rosewood extract used according to the invention being obtained after the separation step (e). According to a particular mode, in step a), the appropriate particle size of the plant material is advantageously obtained by grinding said plant material. The grinding must be as fine as possible to promote the action of the enzymes. Ideally, all particles should be close to 50 pm in size, preferably close to 10 pm.
De préférence, la masse d'eau ajoutée à la matière végétale est égale à 1 à 2 fois la masse de ladite matière végétale et ne dépasse pas cette quantité. Preferably, the mass of water added to the plant material is equal to 1 to 2 times the mass of said plant material and does not exceed this quantity.
En particulier à l’étape b), le rapport entre l'activité de la pectinase et l'activité de la cellulase étant d'au moins 0,14, de préférence compris entre 0,3 et 2,5, et plus préférentiellement entre 0,35 et 0,45, et le rapport entre l'activité de la pectinase et l'activité de l'hémicellulase étant d'au moins 7.10-3 ; de préférence compris entre 1.10-2 et 0,5, et plus préférentiellement entre 1.10-2 et 2.10-2. In particular in step b), the ratio between the pectinase activity and the cellulase activity being at least 0.14, preferably between 0.3 and 2.5, and more preferably between 0.35 and 0.45, and the ratio between the pectinase activity and the hemicellulase activity being at least 7.10 -3 ; preferably between 1.10 -2 and 0.5, and more preferably between 1.10 -2 and 2.10 -2 .
Plus particulièrement, le mélange enzymatique utilisé contient au moins une cellulase, au moins une hemicellulase, au moins une pectinase, et avantageusement en outre au moins une protéase. Selon un mode particulier, la ou les cellulase(s), hemicellulase(s) et pectinase(s) représentent 75% du mélange enzymatique et la ou les protéase(s) représentent 25% dudit mélange enzymatique. Selon un mode particulier, les cellulases sont présentes en une teneur allant de 1 à 50%, de préférence de 20 à 30% en poids du mélange enzymatique, les hémicellulases sont présentes en une teneur allant de 1 à 50%, de préférence de 20 à 30% en poids du mélange enzymatique et les pectinases sont présentes en une teneur allant de 1 à 50%, de préférence de 20 à 30% en poids du mélange enzymatique. Et avantageusement les protéases sont présentes en une teneur allant de 1 à 50%, de préférence de 20 à 30% en poids du mélange enzymatique. More particularly, the enzymatic mixture used contains at least one cellulase, at least one hemicellulase, at least one pectinase, and advantageously also at least one protease. According to a particular mode, the cellulase(s), hemicellulase(s) and pectinase(s) represent 75% of the enzymatic mixture and the protease(s) represent 25% of said enzymatic mixture. According to a particular mode, the cellulases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture, the hemicellulases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture and the pectinases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture. And advantageously the proteases are present in a content ranging from 1 to 50%, preferably from 20 to 30% by weight of the enzymatic mixture.
Dans un mode de réalisation préféré le mélange enzymatique utilisé contient 25% de cellulases, 25% d’hémicellulases, 25% de pectinases et 25% de protéases. In a preferred embodiment, the enzymatic mixture used contains 25% cellulases, 25% hemicellulases, 25% pectinases and 25% proteases.
De préférence, la quantité utilisée du mélange enzymatique tel que défini ci-dessus est comprise entre 0,25% et 10%, de préférence entre 1 % et 6%, en volume du mélange eau/matière végétale. Preferably, the quantity used of the enzymatic mixture as defined above is between 0.25% and 10%, preferably between 1% and 6%, by volume of the water/plant material mixture.
Avantageusement, l'incubation selon l'étape c) est réalisée pendant 2 à 20 heures, de préférence entre 4 et 12 heures, à une température comprise entre 25°C et 75°C, de préférence entre 40°C et 60°C, et de préférence autour de 50°C. La réaction d'hydrolyse est ensuite stoppée par désactivation des enzymes par chauffage de préférence entre 80°C et 105°C pendant par exemple 5 à 20 minutes Advantageously, the incubation according to step c) is carried out for 2 to 20 hours, preferably between 4 and 12 hours, at a temperature between 25°C and 75°C, preferably between 40°C and 60°C. , and preferably around 50°C. The hydrolysis reaction is then stopped by deactivation of the enzymes by heating preferably between 80°C and 105°C for for example 5 to 20 minutes.
Puis le milieu réactionnel est séparé, selon l'étape d), en utilisant toutes les techniques de séparation connues adaptées, telle que la centrifugation ou la décantation. Then the reaction medium is separated, according to step d), using all suitable known separation techniques, such as centrifugation or decantation.
Selon un mode particulier, le mélange est préfiltré sur tamis de 0.8mm puis 0.5mm afin de retirer la partie le plus importante de la phase solide, puis une filtration stérilisante est avantageusement réalisée sur plaques clarifiantes d’une taille de pores de 0.2pm, à une pression de 2 Bars. According to a particular mode, the mixture is pre-filtered through a 0.8mm then 0.5mm sieve in order to remove the most important part of the solid phase, then sterilizing filtration is advantageously carried out on clarifying plates with a pore size of 0.2pm, at a pressure of 2 Bars.
La phase aqueuse obtenue après filtration stérilisante est stabilisée avec les conservateurs (acide citrique, sorbate de potassium, benzoate de sodium). The aqueous phase obtained after sterilizing filtration is stabilized with preservatives (citric acid, potassium sorbate, sodium benzoate).
Selon un mode de réalisation particulier et préféré, la composition cosmétique selon l’invention est caractérisée en ce que l’extrait aqueux de bois de rose est obtenu par un procédé d’extraction enzymatique comprenant les étapes : a) addition d'eau au bois de rose préalablement broyé présentant une taille de particule appropriée, b) addition d'un mélange enzymatique contenant au moins une cellulase, au moins une hémicellulase et au moins une pectinase, et avantageusement en outre au moins une protéase, c) incubation sous agitation du bois de rose préalablement broyé et du mélange enzymatique pour libérer dans le milieu réactionnel des huiles, des protéines et des sucres fermentescibles, pendant une durée dépendant des rendements recherchés, d) séparation du milieu réactionnel pour obtenir de l'huile libre, une phase aqueuse contenant des protéines et des sucres fermentescibles, et une phase solide, e) séparation de la phase aqueuse et ajout éventuel de conservateurs. According to a particular and preferred embodiment, the cosmetic composition according to the invention is characterized in that the aqueous extract of rosewood is obtained by an enzymatic extraction process comprising the steps: a) addition of water to the wood of previously crushed rose having an appropriate particle size, b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously also at least one protease, c) incubation with stirring of the previously crushed rosewood and the enzymatic mixture to release fermentable oils, proteins and sugars into the reaction medium, for a period depending on the desired yields, d) separation of the reaction medium to obtain free oil, an aqueous phase containing fermentable proteins and sugars, and a solid phase, e) separation of the aqueous phase and possible addition of preservatives.
L’extrait aqueux de bois de rose utilisé selon l’invention est caractérisé notamment par la présence d’acides aminés (acide aspartique, tyrosine, arginine) et de sucres totaux (glucose, fructose). The aqueous extract of rosewood used according to the invention is characterized in particular by the presence of amino acids (aspartic acid, tyrosine, arginine) and total sugars (glucose, fructose).
L’extrait aqueux de bois de rose utilisé selon l’invention se caractérise également par le fait qu’il ne s’agit pas d’un extrait fermenté. Selon un mode particulier, l’extrait aqueux de bois de rose utilisé selon l’invention comprend de l’eau en une teneur allant de 94 à 96%, une teneur en extrait sec allant de 1 à 6% et préférentiellement de 3 à 5% et des conservateurs en une teneur allant de 0,5 à 1 % en poids par rapport au poids total de l’extrait. Cet extrait de bois de rose selon l’invention est dénommé en nom INCI water (and) rose extract (and) citric acid (and) potassium sorbate (and) sodium benzoate. The aqueous rosewood extract used according to the invention is also characterized by the fact that it is not a fermented extract. According to a particular mode, the aqueous extract of rosewood used according to the invention comprises water in a content ranging from 94 to 96%, a dry extract content ranging from 1 to 6% and preferably from 3 to 5 % and preservatives in a content ranging from 0.5 to 1% by weight relative to the total weight of the extract. This rosewood extract according to the invention is called INCI water (and) rose extract (and) citric acid (and) potassium sorbate (and) sodium benzoate.
L’extrait aqueux de bois de rose utilisé selon l’invention sera généralement présent en une teneur allant de 0,1 à 1 %, en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition. The aqueous extract of rosewood used according to the invention will generally be present in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
Peptides Peptides
Les compositions de l’invention comprennent également un acetyl hexapeptide et un palmitoyl tetrapeptide, en particulier les peptides suivants : The compositions of the invention also comprise an acetyl hexapeptide and a palmitoyl tetrapeptide, in particular the following peptides:
Acetyl hexapeptide 8 de la société LIPOTEC commercialisé sous la dénomination ARGIRELINE® Amplified peptide de nom INCI : WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE. Acetyl hexapeptide 8 from the company LIPOTEC marketed under the name ARGIRELINE® Amplified peptide with INCI name: WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE.
Selon un mode particulier, le produit commercial ARGIRELINE® Amplified peptide contenant l’acetyl hexapeptide-8 est présent dans la composition de l’invention en une teneur allant de 0,1% à 10% en poids par rapport au poids total de la composition, de préférence de 1 % à 5% en poids par rapport au poids total de la composition. According to a particular mode, the commercial product ARGIRELINE® Amplified peptide containing acetyl hexapeptide-8 is present in the composition of the invention in a content ranging from 0.1% to 10% by weight relative to the total weight of the composition , preferably from 1% to 5% by weight relative to the total weight of the composition.
En particulier l’acetyl hexapeptide-8 est présent dans la composition de l’invention en une teneur en peptide allant de 0,005% à 0,5% en poids par rapport au poids total de la composition, de préférence de 0,05% à 0,25% en poids par rapport au poids total de la composition. In particular, acetyl hexapeptide-8 is present in the composition of the invention in a peptide content ranging from 0.005% to 0.5% by weight relative to the total weight of the composition, preferably from 0.05% to 0.25% by weight relative to the total weight of the composition.
Palmitoyl tetrapeptide-7 qui est un peptide synthétique de séquence Palmitoyl-Gly-GIn-Pro- Arg (Palmitoyl-GQPR) de la société SEDERMA commercialisé sous la dénomination RIGIN® de nom INCI : Glycerin, Lactic Acid and Palmitoyl Tetrapeptide-7. Palmitoyl tetrapeptide-7 which is a synthetic peptide with the sequence Palmitoyl-Gly-GIn-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with the INCI name: Glycerin, Lactic Acid and Palmitoyl Tetrapeptide-7.
Selon un mode particulier, le produit commercial RIGIN® contenant le palmitoyl tetrapeptide- 7 est présent dans la composition de l’invention en une teneur allant de 0,1 % à 10% en poids par rapport au poids total de la composition, de préférence de 2% à 5% en poids par rapport au poids total de la composition. En particulier palmitoyl tetrapeptide-7 est présent dans la composition de l’invention en une teneur en peptide allant de 0,005% à 0,5% en poids par rapport au poids total de la composition, de préférence de 0,1 % à 0,25% en poids par rapport au poids total de la composition. According to a particular mode, the commercial product RIGIN® containing palmitoyl tetrapeptide-7 is present in the composition of the invention in a content ranging from 0.1% to 10% by weight relative to the total weight of the composition, preferably from 2% to 5% by weight relative to the total weight of the composition. In particular palmitoyl tetrapeptide-7 is present in the composition of the invention in a peptide content ranging from 0.005% to 0.5% by weight relative to the total weight of the composition, preferably from 0.1% to 0. 25% by weight relative to the total weight of the composition.
Selon un mode particulier et préféré, la composition de l’invention comprend en outre une fraction bioactive isolée de roses telle que décrite ci-après. According to a particular and preferred embodiment, the composition of the invention further comprises a bioactive fraction isolated from roses as described below.
Ainsi, la composition de l’invention comprendra généralement : Thus, the composition of the invention will generally include:
(i) L’extrait aqueux de bois de rose de la variété Evanrat en une teneur allant de 0,1 à 1 %, en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition, (i) The aqueous extract of rosewood of the Evanrat variety in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition,
(ii) L’hexapeptide de nom INCI : Acetyl Hexapeptide-8 en une teneur allant 0,05% à 0,25% en poids (de peptide) par rapport au poids total de la composition, et(ii) The INCI name hexapeptide: Acetyl Hexapeptide-8 in a content ranging from 0.05% to 0.25% by weight (of peptide) relative to the total weight of the composition, and
(iii) Le tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7 en une teneur allant de 0,1 % à 0,25% en poids (de peptide) par rapport au poids total de la composition, et (iii) The tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7 in a content ranging from 0.1% to 0.25% by weight (of peptide) per relative to the total weight of the composition, and
(iv) La fraction bioactive isolée de roses fraiches de la variété Evanrat, en une teneur allant de 0 à 1 %, notamment de 0,1 à 1% et en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition. (iv) The bioactive fraction isolated from fresh roses of the Evanrat variety, in a content ranging from 0 to 1%, in particular from 0.1 to 1% and in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
Fraction bioactive isolée de roses et procédé de préparation Bioactive fraction isolated from roses and method of preparation
Par « fraction bioactive » ou « fraction bioactive isolée de roses » ou « Sérum Rose De Granville® », « Zêta » ou « Zêta fraction », on entend un extrait de roses de la variété Evanrat, ou rose ‘Jardin de Granville®’, comprenant les enzymes, protéines, sucres, ions et autres molécules actives présentes dans le cytosol des cellules composant les différents tissus végétaux des roses. L’extrait selon l’invention est distinct d’un cryoextrait de pétales roses tel que décrit dans la demande FR3066388. By “bioactive fraction” or “bioactive fraction isolated from roses” or “Sérum Rose De Granville®”, “Zêta” or “Zêta fraction”, we mean an extract of roses of the Evanrat variety, or rose 'Jardin de Granville®' , comprising the enzymes, proteins, sugars, ions and other active molecules present in the cytosol of the cells making up the different plant tissues of roses. The extract according to the invention is distinct from a cryoextract of pink petals as described in application FR3066388.
Matériel végétal Plant material
L’extrait de roses sous la forme d’une fraction bioactive isolée utilisé selon l’invention peut être préparée à partir de roses fraiches de la variété Evanrat, ou roses ‘Jardin de Granville®’, congelées, lyophilisées, ou d’un mélange quelconque de celles-ci. Dans le contexte de l’invention, on utilise de préférence des roses fraiches. The rose extract in the form of an isolated bioactive fraction used according to the invention can be prepared from fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, frozen, freeze-dried, or any mixture thereof. In the context of the invention, fresh roses are preferably used.
Selon un mode particulier, on utilisera des roses d’été pour préparer la fraction bioactive isolée de roses, en particulier des pétales de roses d’été. Selon un autre mode, on utilisera de préférence, des roses d’hiver, en particulier des pétales de roses d’hiver. According to a particular method, summer roses will be used to prepare the bioactive fraction isolated from roses, in particular summer rose petals. According to another method, we will preferably use winter roses, in particular winter rose petals.
Il est particulièrement avantageux d’utiliser les pétales des roses de la variété Evanrat car ils sont riches en sucres monosaccharides (fructose, glucose, saccharose), des acides organiques (acide citrique, acide malique), des polyphénols (catéchine), de la vitamine C, des acides aminés (majoritairement acide aspartique, acide glutamique, asparagine et glutamine), des minéraux (cendres, potassium, calcium), et des caroténoïdes. It is particularly advantageous to use the petals of roses of the Evanrat variety because they are rich in monosaccharide sugars (fructose, glucose, sucrose), organic acids (citric acid, malic acid), polyphenols (catechin), vitamin C, amino acids (mainly aspartic acid, glutamic acid, asparagine and glutamine), minerals (ash, potassium, calcium), and carotenoids.
Par riche en sucres monosaccharides, on entend un extrait de roses sous la forme d’une fraction bioactive isolée, dans lequel ledit extrait comprend au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose. By rich in monosaccharide sugars is meant a rose extract in the form of an isolated bioactive fraction, in which said extract comprises at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose.
Par riche en minéraux, on entend un extrait de roses sous la forme d’une fraction bioactive isolée, dans lequel ledit extrait comprend au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. By rich in minerals, we mean a rose extract in the form of an isolated bioactive fraction, in which said extract comprises at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
Ainsi, selon un mode de réalisation, la composition selon l’invention comprend en outre un extrait de roses sous la forme d’une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. Thus, according to one embodiment, the composition according to the invention further comprises a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
Avantageusement, le calcium améliore la différenciation épidermique et renforce la structure de la peau, tandis que le potassium amplifie l’hydratation cutanée et booste l’assimilation de l’énergie. Les phyto-sucres (fructose, glucose, saccharose) permettent de gorger les cellules cutanées d’énergie. Advantageously, calcium improves epidermal differentiation and strengthens the structure of the skin, while potassium amplifies skin hydration and boosts energy assimilation. Phyto-sugars (fructose, glucose, sucrose) help fill skin cells with energy.
Procédé d’extraction Extraction process
On utilise comme matériel végétal des roses fraiches de la variété Evanrat ou roses ‘Jardin de Granville®’. Selon un mode préféré, il s’agit des pétales frais de roses de la variété Evanrat ou roses ‘Jardin de Granville®’. We use fresh roses of the Evanrat variety or ‘Jardin de Granville®’ roses as plant material. According to a preferred mode, these are fresh rose petals of the Evanrat variety or ‘Jardin de Granville®’ roses.
Avantageusement, ledit procédé ne requiert l’ajout d’aucun solvant ou liquide exogène. Le procédé mis en oeuvre pour obtenir la fraction bioactive isolée utilisée selon l’invention comprend les étapes principales de : a) Nettoyage du matériel végétal, macération, pressage puis séparation mécanique du matériel végétal pour obtenir une dispersion colloïdale intracellulaire (DCI) et un matériau enrichi en fibres (fraction A) ; b) « Traitement A » puis séparation mécanique de la Dispersion Colloïdale Intracellulaire (DCI) pour obtenir le Surnageant A et une Fraction Membranaire (Fraction B) ; c) « Traitement B » puis séparation mécanique du Surnageant A pour obtenir le Surnageant B et la Fraction C (Fraction Cytoplasmique) ; d) « Traitement C » puis séparation mécanique du Surnageant B pour donner une « Fraction sérique bioactive » et une Fraction D (Précipité) ; et e) Optionnellement, mélange de la « fraction sérique bioactive » avec au moins un conservateur et/ou stabilisant. Advantageously, said process does not require the addition of any exogenous solvent or liquid. The process implemented to obtain the isolated bioactive fraction used according to the invention comprises the main steps of: a) Cleaning of the plant material, maceration, pressing then mechanical separation of the plant material to obtain an intracellular colloidal dispersion (ICD) and a material enriched with fiber (fraction A); b) “Treatment A” then mechanical separation of the Intracellular Colloidal Dispersion (ICD) to obtain Supernatant A and a Membrane Fraction (Fraction B); c) “Treatment B” then mechanical separation of Supernatant A to obtain Supernatant B and Fraction C (Cytoplasmic Fraction); d) “Treatment C” then mechanical separation of Supernatant B to give a “Bioactive Serum Fraction” and a Fraction D (Precipitate); and e) Optionally, mixing the “bioactive serum fraction” with at least one preservative and/or stabilizer.
L’extrait de roses fraiches, de préférence de pétales de roses frais, de la variété Evanrat ou rose ‘Jardin de Granville®’ obtenu à l’issue de ce procédé, est une « fraction sérique bioactive » ou « fraction bioactive » au sens de l’invention. The extract of fresh roses, preferably fresh rose petals, of the Evanrat variety or 'Jardin de Granville®' rose obtained at the end of this process, is a "bioactive serum fraction" or "bioactive fraction" in the sense of the invention.
Par « nettoyage » on entend l'élimination des débris des roses de Granville® fraîches, et de préférence des pétales frais, avant un traitement ultérieur, d'une manière qui évite de blesser la plante, ou l'élimination de composants précieux. Par exemple, il peut être effectué par rinçage à basse pression avec de l'eau potable dans des conditions où le lavage à l'eau de ruissellement ne contiendrait pas sensiblement de pigments végétaux. L'excès d'eau de lavage est ensuite éliminé des plantes lavées. “Cleaning” means the removal of debris from fresh Granville® roses, and preferably fresh petals, before further processing, in a manner that avoids injury to the plant, or the removal of valuable components. For example, it can be carried out by low pressure rinsing with potable water under conditions where washing with runoff water would not contain appreciably plant pigments. Excess wash water is then removed from the washed plants.
Par « macération » on entend le fait de transformer les roses de Granville® fraîches, et de préférence les pétales frais, en particules plus petites pour rompre leur intégrité et faciliter par la suite l'expulsion de la dispersion colloïdale intracellulaire (ICD) liquide. Des exemples d'instruments de macération appropriés comprennent, sans s'y limiter, des dispositifs tels qu'un concasseur, une meule ou un broyeur (par exemple, un broyeur à couteaux, un broyeur à marteaux, etc.). Pour éviter une dégradation du matériel végétal induite par la température, l'étape de macération peut inclure une surveillance de la température et la sélection des paramètres de macération garantissant qu'il n'y a pas d'augmentation significative de la température du matériel végétal au cours de cette étape. Par « pressage » on entend la séparation de la matière liquide des roses de Granville® fraîches, et de préférence des pétales frais, par application d'une force mécanique. Cela inclut, mais sans s'y limiter, des techniques telles que le drainage par gravité ambiante, le pressage par un objet lourd, la force centrifuge d'un expulseur rotatif, la pression du piston d'une presse hydraulique, ou des rouleaux ou une vis de type approprié pour une presse. By “maceration” we mean the act of transforming fresh Granville® roses, and preferably fresh petals, into smaller particles to break their integrity and subsequently facilitate the expulsion of the liquid intracellular colloidal dispersion (ICD). Examples of suitable macerating instruments include, but are not limited to, devices such as a crusher, grinding stone, or grinder (e.g., knife mill, hammer mill, etc.). To avoid temperature-induced degradation of the plant material, the maceration step may include temperature monitoring and selection of maceration parameters ensuring that there is no significant increase in the temperature of the plant material during this step. By “pressing” we mean the separation of the liquid material from fresh Granville® roses, and preferably from fresh petals, by application of mechanical force. This includes, but is not limited to, techniques such as ambient gravity drainage, pressing by a heavy object, centrifugal force from a rotating expeller, piston pressure from a hydraulic press, or rollers or a screw of the appropriate type for a press.
Par « matériau enrichi en fibres » ou « MEF » ou « FEM » (Fibre Enriched Material), on entend une fraction solide et/ou semi-solide enrichie en fibres de roses de Granville® fraîches, de préférence de pétales frais, dont la dispersion colloïdale intracellulaire liquide (ICD) a été éliminée par pressage. By “fiber-enriched material” or “MEF” or “FEM” (Fiber Enriched Material), we mean a solid and/or semi-solid fraction enriched with fresh Granville® rose fibers, preferably fresh petals, of which the Liquid intracellular colloid dispersion (ICD) was removed by pressing.
Par « dispersion colloïdale intracellulaire » ou « DCI » ou « ICD » (Intracellular Colloidal Dispersion) on entend la matière liquide expulsée par pressage de roses de Granville® fraîches, de préférence des pétales frais. Le liquide résultant contient des particules solides et/ou semi-solides dispersées et, potentiellement, des gouttelettes de liquides non miscibles à l'eau de diverses tailles (collectivement appelées particules), dans un milieu aqueux contigu. Les particules sont principalement constituées d'organites de cellules végétales, de fragments d'organites et de matières résiduelles enrichies en fibres. Le milieu aqueux est principalement constitué de cytosols et de vacuoles. By “intracellular colloidal dispersion” or “DCI” or “ICD” (Intracellular Colloidal Dispersion) is meant the liquid material expelled by pressing fresh Granville® roses, preferably fresh petals. The resulting liquid contains dispersed solid and/or semi-solid particles and, potentially, water-immiscible liquid droplets of various sizes (collectively referred to as particles), in a contiguous aqueous medium. The particles mainly consist of plant cell organelles, organelle fragments and fiber-enriched residual materials. The aqueous medium is mainly made up of cytosols and vacuoles.
Par « séparation » on entend la séparation de particules solides et/ou semi-solides, et de gouttelettes de liquide non aqueux à partir d'un liquide aqueux en exploitant la densité et/ou la taille des particules. Cela inclut, mais sans s'y limiter, des techniques telles que l’égouttage, la filtration (y compris la filtration utilisant un gradient de pression), l'écumage, la sédimentation par gravité ambiante, la décantation, la centrifugation ou une combinaison de ce qui précède. De préférence, on utilisera la séparation mécanique en flux continu, mais cela n'exclut pas le traitement par lots. « Séparation 1 » et « Séparation 2 » se réfèrent aux étapes respectives du procédé, effectuées avec des paramètres respectifs. By “separation” we mean the separation of solid and/or semi-solid particles, and droplets of non-aqueous liquid from an aqueous liquid by exploiting the density and/or size of the particles. This includes, but is not limited to, techniques such as dewatering, filtration (including filtration using a pressure gradient), skimming, ambient gravity sedimentation, decantation, centrifugation, or a combination of the above. Preferably, continuous flow mechanical separation will be used, but this does not exclude batch processing. “Separation 1” and “Separation 2” refer to the respective steps of the process, carried out with respective parameters.
Par « surnageant », il est fait référence à un matériau aqueux à partir duquel des particules ont été séparées. « Surnageant A » et « Surnageant B » désignent les surnageants résultant des étapes de séparation respectives du procédé. By "supernatant" is meant an aqueous material from which particles have been separated. “Supernatant A” and “Supernatant B” designate the supernatants resulting from the respective separation steps of the process.
Par « précipité », il est fait référence aux particules à partir desquelles un matériau aqueux a été séparé. « Fraction B » et « Fraction C » désignent les précipités résultant des étapes de séparation respectives du procédé. Les termes « fraction de sérum de roses de Granville® fraîches » et « fraction de sérum de pétales frais de roses de Granville® » désignent des compositions produites par le procédé tel que représenté ci-dessus sans conservateurs et/ou stabilisants ajoutés pour protéger la composition de l'ingrédient contre les facteurs environnementaux tels que la température, l'atmosphère (par exemple, l'oxygène), la lumière et les micro-organismes. By "precipitate" refers to particles from which aqueous material has been separated. “Fraction B” and “Fraction C” designate the precipitates resulting from the respective separation steps of the process. The terms “fresh Granville® rose serum fraction” and “fresh Granville® rose petal serum fraction” designate compositions produced by the process as shown above without preservatives and/or stabilizers added to protect the composition of the ingredient against environmental factors such as temperature, atmosphere (e.g., oxygen), light and microorganisms.
Par « conservateurs et/ou stabilisants » on entend des substances qui, lorsqu'elles sont ajoutées à une « fraction de sérum de roses de Granville® fraîches », de préférence une « fraction de sérum de pétales frais de roses de Granville® », la protègent contre les facteurs environnementaux tels que la température, l'atmosphère (par exemple l'oxygène), la lumière et les micro-organismes. Des substances appropriées particulières peuvent comprendre, sans limitation, un conservateur, un stabilisant et/ou un mélange de ceux-ci. By “preservatives and/or stabilizers” we mean substances which, when added to a “fraction of fresh Granville® rose serum”, preferably a “fraction of fresh Granville® rose petal serum”, protect it against environmental factors such as temperature, atmosphere (e.g. oxygen), light and microorganisms. Particular suitable substances may include, without limitation, a preservative, a stabilizer and/or a mixture thereof.
Le terme « sérum de roses de Granville® » ou « extrait de roses de Granville® fraîches » tel qu'utilisé ici signifie une combinaison d'une fraction de sérum de roses de Granville® fraîches et de conservateurs et/ou de stabilisants. The term “Granville® rose serum” or “fresh Granville® rose extract” as used herein means a combination of a fraction of fresh Granville® rose serum and preservatives and/or stabilizers.
Le terme « sérum de pétales de roses de Granville® » ou « extrait de pétales frais de roses de Granville® » tel qu'utilisé ici signifie une combinaison d'une fraction de sérum de pétales frais de roses de Granville® et de conservateurs et/ou de stabilisants. Dans un mode de réalisation préféré, l’extrait utilisé selon l’invention est obtenu par la mise en oeuvre du procédé d’extraction divulgué dans les brevets EP2919757, JP 6130924, CN The term “Granville® rose petal serum” or “Granville® fresh rose petal extract” as used herein means a combination of a fraction of Granville® fresh rose petal serum and preservatives and /or stabilizers. In a preferred embodiment, the extract used according to the invention is obtained by implementing the extraction process disclosed in patents EP2919757, JP 6130924, CN
ZL201380057567.6, EP2491939B1 , CN1929851 B, les brevets américains n° 8 734 861 et n° 7 473 435 ; la demande de brevet US n° 16/078925. ZL201380057567.6, EP2491939B1, CN1929851 B, US Patents No. 8,734,861 and No. 7,473,435; US patent application No. 16/078925.
De préférence, les fleurs de roses (Roses de Granville®) et 4 à 5 cm de tige sont récoltés de manière à éviter le hachage ou l’écrasement de la biomasse collectée pour éviter la perturbation de la structure cellulaire des fleurs. La viabilité des plantes collectées peut être testée à l’aide d’un fluoromètre à chlorophylle multimode OS5p (Opti-Sciences Inc, Hudson, NH, États-Unis). Preferably, rose flowers (Roses de Granville®) and 4 to 5 cm of stem are harvested in such a way as to avoid chopping or crushing the collected biomass to avoid disruption of the cellular structure of the flowers. Collected plants can be tested for viability using an OS5p multi-mode chlorophyll fluorometer (Opti-Sciences Inc, Hudson, NH, USA).
Selon un mode de réalisation particulier les fleurs fraîches vivantes, y compris les pétales, le pistil et l’étamine, ont été retirées de la tige, y compris le sépale et le réceptacle, et emballées dans des sacs de stockage et sont placé(e)s immédiatement à des températures négatives comprises entre -20°C et -80°C, comme par exemple -20°C, -40°C ou -60°C. Les fleurs peuvent ainsi être stockées sur de longues périodes, comme par exemple plusieurs mois, ou plusieurs années, avant d’être utilisées aux fins du procédé d’extraction. Selon un mode de réalisation préféré, les fleurs fraîches vivantes, y compris les pétales, le pistil et l’étamine, sont retirées de la tige, y compris le sépale et le réceptacle, et emballées dans des sacs de stockage et placées dans un stockage à une température comprise entre 0 et 20°C et de préférence à une température comprise entre 2 et 6°C jusqu’à ce que la récolte soit terminée. According to a particular embodiment the live fresh flowers, including the petals, the pistil and the stamen, have been removed from the stem, including the sepal and the receptacle, and packed in storage bags and are placed )s immediately at negative temperatures between -20°C and -80°C, such as -20°C, -40°C or -60°C. The flowers can thus be stored for long periods, such as several months, or several years, before being used for the extraction process. According to a preferred embodiment, the live fresh flowers, including the petals, pistil and stamen, are removed from the stem, including the sepal and receptacle, and packed into storage bags and placed in storage at a temperature between 0 and 20°C and preferably at a temperature between 2 and 6°C until the harvest is complete.
Etape a Step a
Une fois la récolte terminée, les roses, de préférence les pétales de roses, sont immédiatement rincés par pulvérisation avec de l'eau de 10°C à 15°C pendant 0,1 à 0,3 minutes avec un débit de 5 à 6 litres par minute. L'excès d'eau est de préférence éliminé des fleurs rincées en les laissant égoutter pendant au moins 1 minute. Les fleurs rincées peuvent alors subir une macération, un pressage et une séparation par pressage mécanique, à rouleaux, hydrauliques, ou extracteur à jus pour extraire le contenu de la dispersion colloïdale intercellulaire liquide (DC I) du matériau enrichi en fibres (« Fraction A »). After the harvest is completed, the roses, preferably rose petals, are immediately rinsed by spraying with water at 10°C to 15°C for 0.1 to 0.3 minutes with a flow rate of 5 to 6 liters per minute. Excess water is preferably removed from the rinsed flowers by allowing them to drain for at least 1 minute. The rinsed flowers can then undergo maceration, pressing, and separation by mechanical, roller, hydraulic, or juice extractor pressing to extract the contents of the liquid intercellular colloidal dispersion (DC I) from the fiber-enriched material (“Fraction A ").
A l’issue de ces étapes, le rendement de la fraction A est compris entre 30% et 65%, de préférence entre 35% et 60% et de manière encore préférée entre 40 % et 55 % (poids/poids). At the end of these steps, the yield of fraction A is between 30% and 65%, preferably between 35% and 60% and even more preferably between 40% and 55% (weight/weight).
Le DCI comprend typiquement de 2% à 20% de matière sèche, de préférence de 4% à 16% de matière sèche, et de manière encore préférée de 6 % à 12 % de matière sèche. The DCI typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and even more preferably from 6% to 12% dry matter.
A cette étape, le DCI peut être congelé pour stockage. Typiquement, il est congelé à -20°C. At this stage, the ICD can be frozen for storage. Typically, it is frozen at -20°C.
Etape b Step b
Lorsque le DCI a été congelé pour stockage, il doit préalablement être décongelé doucement. Typiquement, il est placé à décongeler à 4°C ou dans de la glace. When the ICD has been frozen for storage, it must first be thawed gently. Typically, it is placed to thaw at 4°C or in ice.
Le « traitement A » est réalisé par un traitement de déstabilisation du DCI avec des ondes électromagnétiques produites à partir de magnétrons fonctionnant à une fréquence comprise entre 2,45 et 5,8 GHz. Les paramètres du traitement de déstabilisation sont fixés pour obtenir la diminution de la valeur de la composante réelle de la constante diélectrique basse fréquence (e'o) d'environ 20 Farads par mètre (F/m) par rapport à sa valeur avant le traitement. Ce traitement dégrade la stabilité du DCI en provoquant l'agglomération et/ou l'agrégation de particules (c'est-à-dire des organites, fragments d'organites, matière fibreuse résiduelle) en assemblages suffisamment grands et stables pour permettre et/ou améliorer la séparation mécanique. En effet, on considère la suspension colloïdale intracellulaire obtenue comme une dispersion colloïdale relativement stable composée d'une phase continue (cytoplasme et contenu de vacuoles) et d'une phase dispersée (organites suspendus et leurs fragments). Selon la théorie de Derjaguin-Laundau-Verwey-Overbeek (DLVO), cette stabilité est maintenue par la somme des forces attractives de van der Waals et des forces répulsives des doubles couches électriques. La barrière énergétique résultant de la force répulsive empêche les particules de la phase dispersée de s'approcher à moins qu'elles n'aient suffisamment d'énergie pour surmonter cette barrière, auquel cas la force d'attraction les mettra en contact (elles adhéreront alors de manière irréversible). La théorie DLVO décrit l'interaction et l'énergie potentielle des particules en fonction de leurs paramètres, de leur distance les unes des autres et des caractéristiques de la phase continue. La modification des valeurs des variables affectant la force répulsive affecte la stabilité de la dispersion. Dans des conditions normales de stabilité colloïdale, une augmentation de l'énergie potentielle à mesure que les particules s'approchent les unes des autres constitue une barrière énergétique potentielle qu'il est impossible de surmonter sans apport énergétique externe. Cette barrière énergétique maintient les particules séparées et la dispersion stable. Les conditions modifiées lors du traitement A, permettent à la force de répulsion de la double couche de diminuer au point qu’il il n'existe plus de barrière énergétique potentielle et que les particules peuvent s'approcher et s'agglomérer librement. “Treatment A” is carried out by a destabilization treatment of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz. The parameters of the destabilization treatment are set to obtain the reduction in the value of the real component of the low frequency dielectric constant (e'o) of approximately 20 Farads per meter (F/m) compared to its value before the treatment . This processing degrades the stability of the ICD by causing agglomeration and/or aggregation of particles (i.e., organelles, organelle fragments, residual fibrous material) into assemblies large and stable enough to allow and/or or improve mechanical separation. Indeed, we consider the intracellular colloidal suspension obtained as a relatively stable colloidal dispersion composed of a continuous phase (cytoplasm and vacuole contents) and a dispersed phase (suspended organelles and their fragments). According to the Derjaguin-Laundau-Verwey-Overbeek (DLVO) theory, this stability is maintained by the sum of the attractive van der Waals forces and the repulsive forces of the electrical double layers. The energy barrier resulting from the repulsive force prevents the particles of the dispersed phase from approaching unless they have enough energy to overcome this barrier, in which case the force of attraction will bring them into contact (they will adhere then irreversibly). DLVO theory describes the interaction and potential energy of particles based on their parameters, their distance from each other and the characteristics of the continuous phase. Changing the values of the variables affecting the repulsive force affects the stability of the dispersion. Under normal conditions of colloidal stability, an increase in potential energy as particles approach each other constitutes a potential energy barrier that cannot be overcome without external energy input. This energy barrier keeps the particles separated and the dispersion stable. The conditions modified during treatment A allow the repulsive force of the double layer to decrease to the point that there is no longer a potential energy barrier and the particles can approach and agglomerate freely.
Le rétablissement des conditions initiales ne rend pas la stabilité, car les particules se sont irréversiblement agglomérées. Elles sont ainsi facilement éliminables par des moyens mécaniques (Koganov et coll., sofw journal 2017). Restoring the initial conditions does not restore stability, because the particles have irreversibly agglomerated. They are therefore easily removed by mechanical means (Koganov et al., sofw journal 2017).
De préférence, à l’issue du « traitement A », on réalise l’étape de séparation mécanique du DCI par centrifugation afin de produire le « Surnageant A » et la « Fraction B ». Preferably, at the end of “treatment A”, the step of mechanical separation of the DCI is carried out by centrifugation in order to produce “Supernatant A” and “Fraction B”.
Typiquement, le "surnageant A" a une turbidité inférieure à environ 100 NTU. Typically, "supernatant A" has a turbidity below about 100 NTU.
La « fraction B » comprend typiquement de 10% à 30% de matière sèche, de préférence de 13% à 27% de matière sèche, et de manière encore préférée environ 15,0 % à 25,0 % de matière sèche. “Fraction B” typically comprises from 10% to 30% dry matter, preferably from 13% to 27% dry matter, and even more preferably approximately 15.0% to 25.0% dry matter.
Etape c Step c
Le « traitement B » est réalisé par ajustement du niveau de pH dans le « surnageant A » par titrage avec, par exemple, un alcali jusqu'à obtenir un pH supérieur à 6, de préférence un pH allant de 6,5 à 7,5. Typiquement, on utilisera comme alkali préféré le carbonate de potassium. “Treatment B” is carried out by adjusting the pH level in the “supernatant A” by titration with, for example, an alkali until a pH greater than 6 is obtained, preferably a pH ranging from 6.5 to 7.5. Typically, potassium carbonate will be used as the preferred alkali.
A l’issue du « traitement B », on réalise de préférence l’étape de séparation mécanique du « Surnageant A » par centrifugation afin de produire le « Surnageant B » et la « Fraction C ». At the end of “treatment B”, the step of mechanical separation of “Supernatant A” is preferably carried out by centrifugation in order to produce “Supernatant B” and “Fraction C”.
La « fraction C » comprend typiquement de 5% à 25% de matière sèche, de préférence de 8% à 22% de matière sèche, et de manière encore préférée environ 10,0% à 20,0% de matière sèche. The “fraction C” typically comprises from 5% to 25% dry matter, preferably from 8% to 22% dry matter, and even more preferably approximately 10.0% to 20.0% dry matter.
Etape d Step d
Le « traitement C » est réalisé par ajustement du niveau de pH dans le « surnageant B », notamment par titrage avec, par exemple, de l'acide jusqu’à obtenir un pH inférieur à 4,5. Typiquement, on utilisera comme acide préféré une solution d’acide citrique. “Treatment C” is carried out by adjusting the pH level in the “supernatant B”, in particular by titration with, for example, acid until a pH lower than 4.5 is obtained. Typically, a solution of citric acid will be used as the preferred acid.
A l’issue du « traitement C », on réalise de préférence l’étape de séparation mécanique du « Surnageant B » par centrifugation afin de produire la « fraction de sérum de roses de Granville® fraîches », de préférence la « fraction de sérum de pétales frais de roses de Granville® » (Extrait Non Conservé) et la « Fraction D ». At the end of “treatment C”, the step of mechanical separation of “Supernatant B” is preferably carried out by centrifugation in order to produce the “fresh Granville® rose serum fraction”, preferably the “serum fraction”. of fresh Granville® rose petals” (Unpreserved Extract) and “Fraction D”.
La « fraction de sérum de pétales frais de roses de Granville® » comprend typiquement de 2% à 20% de matière sèche, de préférence de 4% à 16% de matière sèche, et de manière encore préférée de de 6,0% à 10,0% de matière sèche. The “Granville® fresh rose petal serum fraction” typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and even more preferably from 6.0% to 10.0% dry matter.
Etape e Step e
Selon certains modes de réalisation, la fraction de sérum obtenue à l’issue de l’étape d) est mélangée avec au moins un conservateur ou au moins un stabilisant pour donner un ingrédient fini, ou avec une combinaison de ceux-ci pour donner l'extrait de roses de Granville® fraîche ou « Sérum Rose De Granville® », de préférence l'extrait de pétales frais de roses de Granville® ou « Sérum de pétales de Roses De Granville® ». According to certain embodiments, the serum fraction obtained at the end of step d) is mixed with at least one preservative or at least one stabilizer to give a finished ingredient, or with a combination of these to give the fresh Granville® rose extract or “Granville® Rose Serum”, preferably extract of fresh Granville® rose petals or “Granville® Rose Petal Serum”.
Des agents stabilisants particulièrement appropriés peuvent comprendre, sans limitation, un conservateur, un stabilisant et/ou des mélanges de ceux-ci. Les conservateurs et stabilisants appropriés à utiliser dans la présente invention comprennent, sans s'y limiter, le sorbate de potassium, le benzoate de sodium, le métabisulfite de sodium, la glycérine, le propylène glycol, le dipropylène glycol, le butylène glycol, le pentylène glycol, l'hexylène glycol et le caprylyl glycol. Dans un mode de réalisation particulier, les agents stabilisants peuvent comprendre au moins un conservateur, au moins un stabilisant, au moins un antioxydant ou leurs mélanges. Particularly suitable stabilizing agents may include, without limitation, a preservative, a stabilizer and/or mixtures thereof. Suitable preservatives and stabilizers for use in the present invention include, but are not limited to, potassium sorbate, sodium benzoate, sodium metabisulfite, glycerin, propylene glycol, dipropylene glycol, butylene glycol, pentylene glycol, hexylene glycol and caprylyl glycol. In a particular embodiment, the stabilizing agents can comprise at least one preservative, at least one stabilizer, at least one antioxidant or mixtures thereof.
Ainsi, selon un mode de réalisation, la fraction bioactive isolée de roses et utilisée selon l’invention est obtenue par le procédé comprenant les étapes de : a) nettoyage du matériel végétal, macération, pressage puis séparation mécanique du matériel végétal pour obtenir une dispersion colloïdale intracellulaire (DCI) et un matériau enrichi en fibres (fraction A) ; b) déstabilisation du DCI avec des ondes électromagnétiques puis séparation mécanique de la dispersion colloïdale intracellulaire (DCI) pour obtenir le surnageant A et une Fraction Membranaire (fraction B) ; c) ajustement du niveau de pH dans le surnageant A jusqu'à obtenir un pH supérieur à 6, de préférence un pH allant de 6 à 7, puis séparation mécanique du surnageant A pour obtenir le surnageant B et la fraction C (fraction cytoplasmique) ; d) ajustement du niveau de pH dans le surnageant B jusqu’à obtenir un pH inférieur à 4,5 puis séparation mécanique du surnageant B pour donner une « Fraction sérique bioactive » et une fraction D (précipité) ; et e) Optionnellement, mélange de la « fraction sérique bioactive » avec au moins un conservateur et/ou stabilisant. Thus, according to one embodiment, the bioactive fraction isolated from roses and used according to the invention is obtained by the process comprising the steps of: a) cleaning the plant material, maceration, pressing then mechanical separation of the plant material to obtain a dispersion intracellular colloid (DCI) and a fiber-enriched material (fraction A); b) destabilization of the DCI with electromagnetic waves then mechanical separation of the intracellular colloidal dispersion (DCI) to obtain supernatant A and a Membrane Fraction (fraction B); c) adjustment of the pH level in supernatant A until obtaining a pH greater than 6, preferably a pH ranging from 6 to 7, then mechanical separation of supernatant A to obtain supernatant B and fraction C (cytoplasmic fraction) ; d) adjustment of the pH level in the supernatant B until obtaining a pH lower than 4.5 then mechanical separation of the supernatant B to give a “Bioactive serum fraction” and a fraction D (precipitate); and e) Optionally, mixing the “bioactive serum fraction” with at least one preservative and/or stabilizer.
Selon un mode particulier, la fraction bioactive isolée de roses, de préférence de pétales de roses, utilisée selon l’invention comprend de l’eau en une teneur allant de 90 à 94%, une teneur en extrait sec allant de 6 à 10%. Cette fraction bioactive isolée de roses utilisée selon l’invention est dénommée en nom INCI Rosa Hybrid Flower Extract. According to a particular mode, the bioactive fraction isolated from roses, preferably from rose petals, used according to the invention comprises water in a content ranging from 90 to 94%, a dry extract content ranging from 6 to 10% . This bioactive fraction isolated from roses used according to the invention is called INCI name Rosa Hybrid Flower Extract.
Selon un autre mode particulier, la fraction bioactive isolée de roses, de préférence de pétales de roses, obtenu selon l’invention comprend de l’eau en une teneur allant de 89 à 93%, une teneur en extrait sec allant de 6 à 10% et des conservateurs et/ou stabilisants en une teneur allant de 0,3 à 1 % en poids par rapport au poids total de l’extrait. According to another particular mode, the bioactive fraction isolated from roses, preferably from rose petals, obtained according to the invention comprises water in a content ranging from 89 to 93%, a dry extract content ranging from 6 to 10 % and preservatives and/or stabilizers in a content ranging from 0.3 to 1% by weight relative to the total weight of the extract.
La fraction bioactive isolée de roses pourra être présente dans la composition de l’invention en une teneur allant de 0,1 à 1 % en poids, en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition. The bioactive fraction isolated from roses may be present in the composition of the invention in a content ranging from 0.1 to 1% by weight, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
Ainsi, dans un mode de réalisation particulier, la composition cosmétique selon l’invention comprend : 1 . un extrait aqueux de bois de rose de la variété Evanrat en une teneur allant de 0,1 à 1 %, en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition, Thus, in a particular embodiment, the cosmetic composition according to the invention comprises: 1. an aqueous extract of rosewood of the Evanrat variety in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition,
2. un hexapeptide de nom INCI : Acetyl Hexapeptide-8 en une teneur allant de 0,05% à 0,25% en poids (de peptide) par rapport au poids total de la composition,2. a hexapeptide with the INCI name: Acetyl Hexapeptide-8 in a content ranging from 0.05% to 0.25% by weight (of peptide) relative to the total weight of the composition,
3. un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7 en une teneur allant de 0,1 % à 0,25% en poids (de peptide) par rapport au poids total de la composition, et 3. a tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7 in a content ranging from 0.1% to 0.25% by weight (of peptide) relative to to the total weight of the composition, and
4. une fraction bioactive isolée de roses fraiches de la variété Evanrat, en une teneur allant de 0,1 à 1 %, en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition. 4. a bioactive fraction isolated from fresh roses of the Evanrat variety, in a content ranging from 0.1 to 1%, in particular from 0.2 to 0.5% by weight relative to the total weight of the composition.
Composition cosmétique et galénique Cosmetic and galenic composition
Par « composition cosmétique » on entend toute composition à visée cosmétique, c’est à dire esthétique, pouvant être mise en contact avec les parties superficielles du corps humain et plus particulièrement avec les matières kératiniques, en particulier la peau et/ou les lèvres, en particulier la peau du visage et/ou du cou. By “cosmetic composition” is meant any composition for cosmetic purposes, that is to say aesthetic, which can be brought into contact with the superficial parts of the human body and more particularly with keratin materials, in particular the skin and/or the lips, particularly the skin of the face and/or neck.
Par « milieu physiologiquement acceptable » on entend tout excipient convenant à une utilisation topique, en contact avec les matières kératiniques, sans risque de toxicité, d'incompatibilité, d'instabilité et/ou de réponse allergique. By “physiologically acceptable medium” is meant any excipient suitable for topical use, in contact with keratin materials, without risk of toxicity, incompatibility, instability and/or allergic response.
Le milieu physiologiquement acceptable représente généralement de 1 à 99% en poids, par rapport au poids total de ladite composition. The physiologically acceptable medium generally represents 1 to 99% by weight, relative to the total weight of said composition.
Par « utilisation topique » ou « application topique », on entend une composition destinée à une application sur les matières kératiniques. De fait, une composition orale n’est pas destinée à une application topique sur la peau. By “topical use” or “topical application” is meant a composition intended for application to keratin materials. In fact, an oral composition is not intended for topical application to the skin.
Selon un mode de réalisation particulier, la composition cosmétique selon l’invention est caractérisée en ce qu’il s’agit d’une composition de soin et/ou de maquillage de la peau et/ou des lèvres, en particulier d’une composition de soin de la peau. Une composition pour l’application topique selon l’invention pourra être par exemple sous forme d’une crème, émulsion huile-dans-eau, ou eau-dans-huile ou émulsion multiple, solution, suspension, gel, lait, lotion, sérum, de baume, de stick, ou encore de poudre. According to a particular embodiment, the cosmetic composition according to the invention is characterized in that it is a care and/or makeup composition for the skin and/or lips, in particular a composition skin care. A composition for topical application according to the invention may for example be in the form of a cream, oil-in-water emulsion, or water-in-oil or multiple emulsion, solution, suspension, gel, milk, lotion, serum , balm, stick, or even powder.
Selon un mode particulier, la composition cosmétique de l’invention est sous la forme d’une crème, émulsion huile-dans-eau, ou eau-dans-huile ou émulsion multiple, solution, suspension, gel, lait, lotion, ou sérum. According to a particular embodiment, the cosmetic composition of the invention is in the form of a cream, oil-in-water emulsion, or water-in-oil or multiple emulsion, solution, suspension, gel, milk, lotion, or serum .
Dans un mode de réalisation préféré, ladite composition utilisée selon l’invention est sous la forme d’une crème ou d’un sérum. In a preferred embodiment, said composition used according to the invention is in the form of a cream or a serum.
La phase aqueuse de la composition selon l'invention comprend de l'eau et éventuellement un solvant hydrosoluble. The aqueous phase of the composition according to the invention comprises water and optionally a water-soluble solvent.
On entend par ‘solvant hydrosoluble’ selon l’invention, un composé liquide à température ambiante et miscible à l'eau (miscibilité dans l'eau supérieure à 50 % en poids à 25 °C et pression atmosphérique). On peut citer notamment : les mono-alcools inférieurs en C1 -C5 tels que l'éthanol, l'isopropanol et leurs mélanges; les glycols en C2-C8 tels que l'éthylène glycol, le propylène glycol, le 1 ,3-butylène glycol, le dipropylène glycol, et leurs mélanges ; les polyols en C2-C32 tels que les polyglycérols, les polyéthylènes glycols, et leurs mélanges, et leurs mélanges. By 'water-soluble solvent' according to the invention is meant a compound that is liquid at room temperature and miscible with water (miscibility in water greater than 50% by weight at 25°C and atmospheric pressure). We can cite in particular: lower C1 -C5 mono-alcohols such as ethanol, isopropanol and their mixtures; C2-C8 glycols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, dipropylene glycol, and mixtures thereof; C2-C32 polyols such as polyglycerols, polyethylene glycols, and mixtures thereof, and mixtures thereof.
Elle peut comprendre également des gélifiants hydrophiles, des antioxydants, des conservateurs et leurs mélanges. It may also include hydrophilic gelling agents, antioxidants, preservatives and mixtures thereof.
La composition cosmétique selon l’invention peut comprendre en outre une phase grasse (corps gras solides) ou huileuse. The cosmetic composition according to the invention may also comprise a fatty phase (solid fatty substances) or oily phase.
On entend par « phase huileuse » une huile ou un mélange d'huiles miscibles entre elles, ou non. Par « huile », on entend, au sens de l'invention, un corps gras, non soluble dans l'eau, liquide à 25°C et pression atmosphérique. Ces huiles peuvent être volatiles ou non volatiles, végétale, minérale ou synthétique. By “oil phase” is meant an oil or a mixture of oils which may or may not be miscible with each other. By “oil” is meant, within the meaning of the invention, a fatty substance, not soluble in water, liquid at 25°C and atmospheric pressure. These oils can be volatile or non-volatile, vegetable, mineral or synthetic.
Une phase huileuse selon l’invention peut comprendre des huiles naturelles, hydrocarbonées, siliconées, et leurs mélanges. La teneur en phase grasse ou huileuse dans la composition cosmétique de l’invention ira généralement de 0,2 % à 45 %, de préférence de 0,5 % à 30 %, et de préférence encore de 2 % à 25 % en poids par rapport au poids total de ladite composition. An oily phase according to the invention may comprise natural, hydrocarbon, silicone oils, and mixtures thereof. The content of fatty or oily phase in the cosmetic composition of the invention will generally range from 0.2% to 45%, preferably from 0.5% to 30%, and more preferably from 2% to 25% by weight per relative to the total weight of said composition.
La composition de l'invention peut également comprendre tout additif usuellement utilisé en cosmétique tels que des antioxydants, des parfums, des agents actifs cosmétiques, comme par exemple des agents émollients, des agents hydratants, des vitamines, des agents antiâge, des agents liftants, des agents tenseurs, des agents repulpants, des agents éclaircissants, des charges, des nacres et leurs mélanges. The composition of the invention may also comprise any additive usually used in cosmetics such as antioxidants, perfumes, cosmetic active agents, such as for example emollient agents, moisturizing agents, vitamins, anti-aging agents, lifting agents, tightening agents, plumping agents, lightening agents, fillers, pearls and their mixtures.
Ainsi selon un mode de réalisation particulier, l’invention porte sur une composition cosmétique pour application topique sur la peau et/ou les lèvres, en particulier la peau du visage et/ou du corps, comprenant dans un milieu physiologiquement acceptable, au moins un extrait aqueux de bois de rose, de l’Acetyl Hexapeptide-8, du Palmitoyl tétrapeptide-7 et éventuellement une fraction bioactive isolée obtenue à partir de rose fraiche, et au moins un adjuvant cosmétique choisi dans le groupe constitué par les antioxydants, les parfums, les agents émollients, les agents hydratants, les vitamines, les agents anti-âge, les agents liftants, les agents tenseurs, les agents repulpants, les agents éclaircissants, les charges, les nacres et leurs mélanges. Thus according to a particular embodiment, the invention relates to a cosmetic composition for topical application to the skin and/or lips, in particular the skin of the face and/or body, comprising in a physiologically acceptable medium, at least one aqueous extract of rosewood, Acetyl Hexapeptide-8, Palmitoyl tetrapeptide-7 and optionally an isolated bioactive fraction obtained from fresh rose, and at least one cosmetic adjuvant chosen from the group consisting of antioxidants, perfumes , emollient agents, moisturizing agents, vitamins, anti-aging agents, lifting agents, tightening agents, plumping agents, lightening agents, fillers, pearls and mixtures thereof.
Procédé cosmétique Cosmetic process
L’invention porte aussi sur un procédé cosmétique non thérapeutique de soin et/ou de maquillage de la peau et/ou des lèvres comprenant l’application sur la peau et/ou les lèvres, en particulier la peau, d’une composition telle que définie selon l’invention. The invention also relates to a non-therapeutic cosmetic process for caring for and/or making up the skin and/or lips comprising the application to the skin and/or lips, in particular the skin, of a composition such as defined according to the invention.
Par « peau et/ou lèvres » selon l’invention, on entend notamment une peau et/ou des lèvres saines, c’est-à-dire ne présentant pas de troubles ou de désordres qui relèveraient d’un état pathologique (sujets ‘non sains’, atteints d’une pathologie). By “skin and/or lips” according to the invention, we mean in particular healthy skin and/or lips, that is to say not presenting disorders or disorders which would relate to a pathological state (subjects ' unhealthy', suffering from a pathology).
Selon un mode particulier, l’application de la composition telle que définie selon l’invention sur la peau et/ou les lèvres permet de prévenir et/ou diminuer les signes du vieillissement cutané, en particulier la perte de densité et/ou de fermeté de la peau, la diminution de l’épaisseur de l’épiderme, la diminution de la différenciation et/ou prolifération des cellules épidermiques, et/ou l’apparition de rides et/ou ridules. Association d’actifs et utilisations According to a particular mode, the application of the composition as defined according to the invention to the skin and/or lips makes it possible to prevent and/or reduce the signs of skin aging, in particular the loss of density and/or firmness. of the skin, the reduction in the thickness of the epidermis, the reduction in the differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines. Association of assets and uses
La présente invention concerne aussi une association d’actifs cosmétiques comprenant : The present invention also relates to a combination of cosmetic active ingredients comprising:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8,
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7, et (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, and
(iv) Eventuellement en outre une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. (iv) Optionally also an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
L’invention porte également sur l’utilisation cosmétique non thérapeutique de l’association d’actifs cosmétiques telle que définie ci-dessus, comme association active pour prévenir et/ou diminuer les signes du vieillissement cutané, en particulier la perte de densité et/ou de fermeté de la peau, la diminution du de l’épaisseur de l’épiderme, la diminution de la différenciation et/ou prolifération des cellules épidermiques, et/ou l’apparition de rides et/ou ridules. The invention also relates to the non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined above, as an active association for preventing and/or reducing the signs of skin aging, in particular loss of density and/or or firmness of the skin, the reduction in the thickness of the epidermis, the reduction in the differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines.
Ensemble cosmétique et utilisation Cosmetic set and use
La présente invention concerne aussi un ensemble cosmétique comprenant : The present invention also relates to a cosmetic set comprising:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8,
(iii) Un tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7, et (iii) A tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and
(iv) Eventuellement en outre une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium, caractérisé en ce que l’un au moins des actifs (i) à (iv) est conditionné dans une composition distincte des autres actifs, et en particulier chaque actif est conditionné dans une composition distincte. (iv) Optionally also an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium, characterized in that at least one of the active ingredients ( i) to (iv) is packaged in a composition distinct from the other active ingredients, and in particular each active ingredient is packaged in a distinct composition.
Un autre objet est un procédé cosmétique comprenant l’application successive, sur la peau et/ou les lèvres, des compositions de l’ensemble cosmétique tel que défini ci-dessus. Another object is a cosmetic process comprising the successive application, to the skin and/or lips, of the compositions of the cosmetic set as defined above.
L’invention va désormais être illustrée dans les exemples non limitatifs suivants. Sauf indication contraire, les pourcentages (%) sont exprimés en pourcentages (%) en poids par rapport au poids total de la composition. The invention will now be illustrated in the following non-limiting examples. Unless otherwise indicated, percentages (%) are expressed as percentages (%) by weight relative to the total weight of the composition.
EXEMPLES EXAMPLES
Exemple 1 : Matériels et méthodes 1.1 Préparation des cultures cellulaires Example 1: Materials and methods 1.1 Preparation of cell cultures
Les différents tests des ingrédients seuls ou en combinaison selon l’invention ont été effectués sur des échantillons de peau reconstruite par bioimpression par extrusion avec des cellules issues d’un donneur d’âge mature (= 42 ans). Cellules utilisées The various tests of the ingredients alone or in combination according to the invention were carried out on samples of skin reconstructed by bioprinting by extrusion with cells from a donor of mature age (= 42 years). Cells used
L’ensemble des cellules utilisées a été extrait par LabSkin Creations à partir de résidus opératoires de donneurs ayant subi une chirurgie plastie ou réparatrice. Le recueil de ces résidus opératoires a été obtenu selon la réglementation Française en vigueur et autorisée par le Ministère de la Recherche sous le numéro DC2020-4346. Afin de garantir leur utilisation en ingénierie tissulaire, des banques de cellules primaires ont été réalisées à différents passages selon le type cellulaire considéré. Les cellules ont été sécurisées d’un point de vue microbiologique et sont exempts de mycoplasmes. [Table 1]
Figure imgf000025_0001
Figure imgf000026_0001
All of the cells used were extracted by LabSkin Creations from surgical residues from donors who underwent plastic or reconstructive surgery. The collection of these operational residues was obtained according to current French regulations and authorized by the Ministry of Research under number DC2020-4346. In order to guarantee their use in tissue engineering, primary cell banks were produced at different passages depending on the cell type considered. The cells have been microbiologically safe and are free of mycoplasma. [Table 1]
Figure imgf000025_0001
Figure imgf000026_0001
Bioimpression et culture des peaux reconstruites Bioprinting and culture of reconstructed skins
Bioimpression des dermes Dermis bioprinting
Les fibroblastes issus de donneurs âgés ont été trypsinés et comptés. Les cellules ont été mises en suspension dans la bioencre constituée d’alginate, gélatine et fibrinogène (LabSkin Creations) à raison de 250,000 cellules par mL d’encre. Les bio-encres ont été transférées dans des seringues stériles montées sur des canules de 10 gauges. Fibroblasts from elderly donors were trypsinized and counted. The cells were suspended in the bioink consisting of alginate, gelatin and fibrinogen (LabSkin Creations) at a rate of 250,000 cells per mL of ink. The bioinks were transferred into sterile syringes mounted on 10 gauge cannulas.
Les objets imprimés ont été modélisés avec le logiciel Slic3r (GNU Affero General Public License) et Repetier (Hot-World GmbH & Co. KG). Les dermes ont été imprimés grâce à une imprimante 3D (LabSkin Creations) selon une taille de 1.5cm x 1.5cm et comportaient deux couches de cellules extrudées dans la bioencre. The printed objects were modeled with the software Slic3r (GNU Affero General Public License) and Repetier (Hot-World GmbH & Co. KG). The dermis were printed using a 3D printer (LabSkin Creations) to a size of 1.5cm x 1.5cm and had two layers of cells extruded into the bioink.
Les construits ont ensuite été polymérisés 1 heure dans une solution de calcium et thrombine (LabSkin Creations) permettant la consolidation de la bioencre. Après trois rinçages dans un tampon salin, les dermes équivalents ont été mis en culture dans du contenant du DMEM supplémenté de 10% de sérum de veau et de 1% d’antibiotiques et antifongique et placés à l’incubateur (37°C, 5% CO2) pendant 12 jours. Le milieu de culture a été renouvelé tous les 2 jours. The constructs were then polymerized for 1 hour in a solution of calcium and thrombin (LabSkin Creations) allowing the consolidation of the bioink. After three rinses in a saline buffer, the equivalent dermis were cultured in a container of DMEM supplemented with 10% calf serum and 1% antibiotics and antifungal and placed in the incubator (37°C, 5 % CO2) for 12 days. The culture medium was renewed every 2 days.
Épidermisation Epidermization
Les kératinocytes à confluence ont été trypsinés puis comptés. L’équivalent de 500,000 cellules a été ensemencé à la surface de chaque derme bioimprimé. Après 30 minutes d’adhésion à 37°C, du milieu de culture contenant du DMEM supplémenté de 10% de sérum de veau foetal, d’hydrocortisone et d’insuline a été ajouté pour submerger les construits dermo-épidermiques bioimprimés. Confluent keratinocytes were trypsinized and then counted. The equivalent of 500,000 cells was seeded on the surface of each bioprinted dermis. After 30 minutes of adhesion at 37°C, culture medium containing DMEM supplemented with 10% fetal bovine serum, hydrocortisone and insulin was added to submerge the bioprinted dermo-epidermal constructs.
Maturation et différenciation Maturation and differentiation
Les ensembles dermo-épidermiques bioimprimés ont été cultivés durant 7 jours dans du milieu de culture contenant du DMEM supplémenté de 10% de sérum de veau foetal, d’hydrocortisone et d’insuline permettant la prolifération des kératinocytes. Ils ont ensuite été élevés à l’interface air/liquide afin d’initier la différenciation des épidermes. Cette ultime étape de culture de 7 jours a permis d’obtenir des épidermes pluristratifiés. The bioprinted dermo-epidermal assemblies were cultured for 7 days in culture medium containing DMEM supplemented with 10% fetal calf serum, hydrocortisone and insulin allowing the proliferation of keratinocytes. They were then raised at the air/liquid interface in order to initiate the differentiation of the epidermis. This final 7-day culture step made it possible to obtain multistratified epidermis.
Traitements Treatments
Les traitements par les ingrédients actifs seuls ou en combinaisons ont été dissous dans les milieux de culture. Les concentrations finales en ingrédients actifs sont précisées pour chaque exemple. Treatments with active ingredients alone or in combinations were dissolved in the culture media. The final concentrations of active ingredients are specified for each example.
Préparation de l’extrait de bois de rose : Preparation of rosewood extract:
Les bois de rose de la variété Evanrat, en particulier de la variété rose Jardin de Granville®, éventuellement additionnés de fleurs de rose, ont été réduites en poudre en trois étapes. Rosewood of the Evanrat variety, in particular of the Jardin de Granville® rose variety, possibly with the addition of rose flowers, was reduced to powder in three stages.
Etape 1 : passage dans un broyeur de jardin afin d’obtenir des tronçons de deux centimètres Etape 2 : passage dans un broyeur industriel Step 1: passage through a garden shredder to obtain sections of two centimeters Step 2: passage through an industrial shredder
Mise en solution Dissolution
L’extraction enzymatique a été réalisée dans un réacteur d'une contenance maximale de 70I. Le réacteur est agité par hélice et est thermostaté par double enveloppe. The enzymatic extraction was carried out in a reactor with a maximum capacity of 70 I. The reactor is stirred by propeller and is thermostatically controlled by a double jacket.
16Kg de poudre de bois de rose à 0.1 % de fleurs de rose sont dispersés et mis en suspension dans 47,5Kg d'eau. 16Kg of rosewood powder with 0.1% rose flowers are dispersed and suspended in 47.5Kg of water.
Afin de désactiver les enzymes endogènes, le mélange a été élevé à une température de 85°C pendant 1 min puis stabilisé à 50°C. To deactivate endogenous enzymes, the mixture was raised to a temperature of 85°C for 1 min and then stabilized at 50°C.
Lyse des parois végétales Lysis of plant walls
0.8 Kg d’un cocktail Enzymatique HEL1 PR1 comprenant un mélange de cellulases, d’hémicellulases et de pectinases à 75% et des protéases à 25%, ont été ajoutés au mélange réactionnel correspondant (5% w/w). La réaction est poursuivie pendant 4 heures, la température est maintenue à 50°C. 0.8 Kg of an HEL1 PR1 Enzymatic cocktail comprising a mixture of cellulases, hemicellulases and pectinases at 75% and proteases at 25%, were added to the corresponding reaction mixture (5% w/w). The reaction is continued for 4 hours, the temperature is maintained at 50°C.
Inactivation des enzymes Inactivation of enzymes
Le mélange a été porté à 85°C pendant 1 min afin d'inactiver les enzymes, puis ramené à 35°C. The mixture was heated to 85°C for 1 min to inactivate the enzymes, then returned to 35°C.
Pré filtration Pre filtration
Le mélange est préfiltré sur tamis de 0.8mm puis 0.5mm afin de retirer la partie le plus importante de la phase solide, puis sur un drap. Filtration stérilisante The mixture is pre-filtered through a 0.8mm then 0.5mm sieve in order to remove the most important part of the solid phase, then on a sheet. Sterilizing filtration
Une filtration a été réalisée sur plaques clarifiantes (PALL corporation) d’une taille de pores de 0.2pm, à une pression de 2 Bars. Filtration was carried out on clarifying plates (PALL corporation) with a pore size of 0.2 pm, at a pressure of 2 Bars.
Formulation Formulation
La phase aqueuse obtenue après filtration stérilisante est stabilisée avec les conservateurs suivants : The aqueous phase obtained after sterilizing filtration is stabilized with the following preservatives:
- Acide citrique, - Citric acid,
- Sorbate de potassium et - Potassium sorbate and
- Benzoate de sodium. - Sodium benzoate.
On obtient, après filtration, un extrait aqueux de bois de rose à 3.7% en poids de matière sèche par rapport au poids total de l’extrait. After filtration, an aqueous extract of rosewood is obtained with 3.7% by weight of dry matter relative to the total weight of the extract.
Préparation de la fraction active de roses fraîches (ZETA fraction) : Preparation of the active fraction of fresh roses (ZETA fraction):
On utilise comme matériel végétal des pétales frais de roses de la variété Evanrat ou rose « Jardin de Granville® ». We use fresh rose petals of the Evanrat variety or “Jardin de Granville®” rose as plant material.
La fraction bioactive de pétales de rose utilisée selon l’invention est obtenue selon le protocole suivant : a1 ) nettoyage des pétales frais de roses par pulvérisation avec de l'eau d’une température d’environ 12°C, pendant 0,1 à 0,3 minutes à un débit de 5 à 6 litres par minute, a2) macération, pressage puis séparation mécanique des roses à l'aide d'une presse à vis mécanique (modèle CP-6 Vincent Corporation, FL) pour extraire le contenu de la dispersion colloïdale intercellulaire liquide (DCI) du matériau enrichi en fibres (« Fraction A ») ; b1 ) déstabilisation du DCI avec des ondes électromagnétiques produites à partir de magnétrons fonctionnant à une fréquence comprise entre 2,45 et 5,8 GHz, b2) centrifugation de la dispersion colloïdale intracellulaire (DCI) pour obtenir le surnageant A et une Fraction Membranaire (fraction B) ; c1 ) ajustement du niveau de pH dans le surnageant A par un alkali (Le., le carbonate de potassium), afin d’obtenir un pH allant de 6 à 7, c2) séparation mécanique du surnageant A pour obtenir le surnageant B et la fraction C (fraction cytoplasmique) ; d1 ) ajustement du niveau de pH dans le surnageant B par de l’acide citrique afin d’obtenir un pH inférieur à 4,5, d2) séparation mécanique du surnageant B pour donner une « fraction de sérum de pétales frais de roses de Granville® » et une fraction D (précipité) ; et e) mélange de la « fraction de sérum de pétales frais de roses de Granville® » avec du sorbate de potassium et du benzoate de sodium. The bioactive fraction of rose petals used according to the invention is obtained according to the following protocol: a1) cleaning the fresh rose petals by spraying with water at a temperature of approximately 12°C, for 0.1 to 0.3 minutes at a flow rate of 5 to 6 liters per minute, a2) maceration, pressing then mechanical separation of the roses using a mechanical screw press (model CP-6 Vincent Corporation, FL) to extract the contents the liquid intercellular colloidal dispersion (DCI) of the fiber-enriched material (“Fraction A”); b1) destabilization of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz, b2) centrifugation of the intracellular colloidal dispersion (DCI) to obtain the supernatant A and a Membrane Fraction ( fraction B); c1) adjustment of the pH level in the supernatant A using an alkali (Le., potassium carbonate), in order to obtain a pH ranging from 6 to 7, c2) mechanical separation of the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction); d1) adjustment of the pH level in the supernatant B with citric acid in order to obtain a pH lower than 4.5, d2) mechanical separation of the supernatant B to give a “fraction of fresh Granville® rose petal serum” and a fraction D (precipitate); and e) mixing “Granville® fresh rose petal serum fraction” with potassium sorbate and sodium benzoate.
Le sérum de pétales de roses de Granville® (aussi appelé « Zêta sérum », ou « Zf sérum » ou « ZETA FRACTION » ou « ZETA fraction » dans les exemples suivants) contient 8% matière sèche (matière active), 91 ,5% en poids d’eau, 0,15% en poids de sorbate de potassium et 0,3% en poids de benzoate de sodium. Ces teneurs s’entendent en en poids par rapport au poids total de l’extrait. Le nom INCI de cet extrait est Rosa Hybrid Flower Extract, Potassium Sorbate, and Sodium Benzoate. Granville® rose petal serum (also called “Zeta serum”, or “Zf serum” or “ZETA FRACTION” or “ZETA fraction” in the following examples) contains 8% dry matter (active ingredient), 91.5 % by weight water, 0.15% by weight potassium sorbate and 0.3% by weight sodium benzoate. These contents are by weight relative to the total weight of the extract. The INCI name for this extract is Rosa Hybrid Flower Extract, Potassium Sorbate, and Sodium Benzoate.
Acetyl hexapeptide 8 de la société LIPOTEC est commercialisé sous la dénomination ARGIRELINE® Amplified peptide de nom INCI : WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE. Dans les exemples, on entend par « Acetyl hexapeptide 8 » le produit commercial ARGIRELINE® Amplified peptide comprenant 0,05% de peptide. Acetyl hexapeptide 8 from the company LIPOTEC is marketed under the name ARGIRELINE® Amplified peptide with INCI name: WATER and ACETYL HEXAPEPTIDE-8 and SODIUM BENZOATE. In the examples, “Acetyl hexapeptide 8” means the commercial product ARGIRELINE® Amplified peptide comprising 0.05% peptide.
Les pourcentages indiqués dans les exemples correspondent aux pourcentages en poids de produit commercial dans les milieux de culture ou dans les formules cosmétiques. The percentages indicated in the examples correspond to the percentages by weight of commercial product in culture media or in cosmetic formulas.
Palmitoyl tetrapeptide-7 est un peptide synthétique de séquence Palmitoyl-Gly-GIn-Pro-Arg (Palmitoyl-GQPR) de la société SEDERMA commercialisé sous la dénomination RIGIN® de nom INCI : Glycerin, Lactic Acis and Palmitoyl Tetrapeptide-7. Dans les exemples, on entend par « Palmitoyl tetrapeptide-7» le produit commercial RIGIN® comprenant 0,05% de peptide. Les pourcentages indiqués dans les exemples correspondent aux pourcentages en poids de produit commercial dans les milieux de culture ou dans les formules cosmétiques. Palmitoyl tetrapeptide-7 is a synthetic peptide with the sequence Palmitoyl-Gly-GIn-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with the INCI name: Glycerin, Lactic Acis and Palmitoyl Tetrapeptide-7. In the examples, “Palmitoyl tetrapeptide-7” means the commercial product RIGIN® comprising 0.05% peptide. The percentages indicated in the examples correspond to the percentages by weight of commercial product in culture media or in cosmetic formulas.
Les concentrations des actifs ou des combinaisons ont été sélectionnées sur la base des résultats de cytotoxicité pré-effectué. The concentrations of active ingredients or combinations were selected based on the pre-performed cytotoxicity results.
Les peaux reconstruites ont été traitées pendant la reconstruction dermique, de J5 à J 12 et épidermique de J14 à J18 puis de J20 à J26. The reconstructed skins were treated during dermal reconstruction, from D5 to D 12 and epidermal from D14 to D18 then from D20 to D26.
Arrêt des cultures Stopping cultivation
La culture des échantillons des 12 conditions a été arrêtée au jour 26. La moitié des échantillons a été fixée en solution de formol tamponné 4% (Alphapath, France) puis inclus en paraffine. 1.2 Méthodes d’analyse des échantillons de peau reconstruite par bioimpression après tests The culture of samples from the 12 conditions was stopped on day 26. Half of the samples were fixed in 4% buffered formalin solution (Alphapath, France) then embedded in paraffin. 1.2 Methods for analyzing skin samples reconstructed by bioprinting after testing
Les différents traitements (ingrédients actifs seuls ou en combinaison) ont été testés sur les échantillons de peau reconstruite par bioimpression selon l’exemple 1. Différentes analyses ont été effectuées sur ces échantillons de peau reconstruite par bioimpression pour déterminer l’effet de chaque traitement sur la peau. The different treatments (active ingredients alone or in combination) were tested on the skin samples reconstructed by bioprinting according to Example 1. Different analyzes were carried out on these skin samples reconstructed by bioprinting to determine the effect of each treatment on the skin.
Coloration Hématoxyline-Phloxine- Safran Hematoxylin-Phloxine-Saffron coloring
Des coupes paraffine de 5 pm ont été réalisées sur les différents échantillons de peau reconstruite par bioimpression. Après déparaffinage et réhydratation, les échantillons ont été colorés à l’Hématoxyline, Phloxine et Safran (HPS) par trois bains successifs de 5 min. Après rinçage, les coupes ont été déshydratées avant d’être montées sur lamelle avec un milieu de montage hydrophobe à base de xylène. Paraffin sections of 5 μm were made on the different skin samples reconstructed by bioprinting. After deparaffinization and rehydration, the samples were stained with Hematoxylin, Phloxine and Saffron (HPS) in three successive 5 min baths. After rinsing, the sections were dehydrated before being mounted on coverslips with a hydrophobic xylene-based mounting medium.
Immunofluorescence sur coupes paraffine Immunofluorescence on paraffin sections
L’ensemble des marquages a été réalisé sur des coupes de 5pm d’épaisseur. Après un démasquage à la chaleur, une étape de saturation en PBS/BSA 5% a permis de bloquer les sites antigèniques aspécifiques. Les cellules ont ensuite été incubées sur la nuit avec un anticorps primaire spécifique de l’antigène d’intérêt : All markings were made on sections 5 pm thick. After unmasking with heat, a saturation step in PBS/BSA 5% made it possible to block the non-specific antigenic sites. The cells were then incubated overnight with a primary antibody specific for the antigen of interest:
- Anticorps anti Ki-67 : 1/50ème monoclonal, souris, Dako ; - Anti-Ki-67 antibody: 1/50th monoclonal, mouse, Dako;
- Anticorps anti Collagène VII : 1/50ème, monoclonal, souris, Santa Cruz. - Anti-Collagen VII antibody: 1/50th, monoclonal, mouse, Santa Cruz.
Le lendemain, après des étapes de rinçages, les coupes ont été incubées avec un anticorps secondaire Alexa-Fluor 568 anti-souris dilué au 1/1000ème (Thermo Fisher Scientific) et le colorant nucléaire Hoechst (Thermo Fisher Scientific). Un témoin négatif sans anticorps primaire a été réalisé en parallèle. The next day, after rinsing steps, the sections were incubated with an anti-mouse Alexa-Fluor 568 secondary antibody diluted 1/1000 (Thermo Fisher Scientific) and Hoechst nuclear dye (Thermo Fisher Scientific). A negative control without primary antibody was carried out in parallel.
Acquisition des images Image acquisition
Les colorations histologiques et marquages immunohistologiques ont été observés avec le système microscope optique Axio Observer D1 / caméra haute résolution Axiocam (Zeiss, Le Pecq, France). Les images ont été acquises à l’aide du logiciel ZEN pro 2012 et sauvegardées en format « tif » (haute résolution). Histological staining and immunohistological markings were observed with the Axio Observer D1 optical microscope / Axiocam high-resolution camera system (Zeiss, Le Pecq, France). Images were acquired using ZEN pro 2012 software and saved in “tif” format (high resolution).
Analyse d’images Image analysis
L’analyse des images a été réalisée avec le logiciel Imaged (Rasband, W.S., Imaged, U. S. National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997-2017). Pour chaque paramètre une analyse statistique a été réalisée avec un test Student, qui est significatif si sa valeur est inférieure à 0,05. Les résultats sont indiqués selon : * P < 0.05, ** P < 0.01 et *** P < 0.001. Image analysis was carried out with Imaged software (Rasband, WS, Imaged, US National Institutes of Health, Bethesda, Maryland, USA, http://imagej.nih.gov/ij/, 1997-2017). For each parameter a statistical analysis was carried out with a Student test, which is significant if its value is less than 0.05. The results are indicated according to: * P < 0.05, ** P < 0.01 and *** P < 0.001.
Mesure épaisseur Thickness measurement
L’épaisseur des couches cohésives de l’épiderme est mesurée à l’aide d’une carte de distance euclidienne. Les pixels correspondant à l’épiderme sont segmentés des autres pixels de l’image. L’image segmentée est ensuite binarisée puis convertie en carte de distance euclidienne. La membrane basale de l’épiderme est sélectionnée et appliquée sur la carte de distance pour mesurer en chaque point la distance entre la membrane basale aux couches terminales de l’épiderme. La valeur obtenue correspond alors à la distance moyenne entre la membrane basale et les couches terminales de l’épiderme. 9 images sont analysées pour chaque condition. The thickness of the cohesive layers of the epidermis is measured using a Euclidean distance map. The pixels corresponding to the epidermis are segmented from the other pixels of the image. The segmented image is then binarized and converted into a Euclidean distance map. The basement membrane of the epidermis is selected and applied to the distance map to measure at each point the distance between the basement membrane and the terminal layers of the epidermis. The value obtained then corresponds to the average distance between the basement membrane and the terminal layers of the epidermis. 9 images are analyzed for each condition.
Exemple 2 : Effets des actifs et combinaisons selon l’invention sur le facteur KI67 L’effet des actifs seuls ou en combinaisons selon l’invention, sur la production du facteur KI67 par les échantillons de peau reconstruite par bioimpression a été analysé. Chaque test a été effectué en triplicate. Example 2: Effects of the active ingredients and combinations according to the invention on the KI67 factor The effect of the active ingredients alone or in combinations according to the invention on the production of the KI67 factor by the skin samples reconstructed by bioprinting was analyzed. Each test was performed in triplicate.
Les échantillons de peau reconstruite par bioimpression ont été testés respectivement avec les actifs et combinaisons suivantes (dissous dans les milieux de culture) : l’extrait aqueux de bois de rose seul (concentration finale 0,07%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,62pg/ml) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,19%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,62pg/ml ) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,19%) + l’extrait aqueux de bois de rose (concentration finale 0,025%). The skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations (dissolved in the culture media): the aqueous extract of rosewood alone (final concentration 0.07%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%) + aqueous extract of rosewood (final concentration 0.025%).
Un témoin négatif non traité a été effectué, sans composition selon l’invention. An untreated negative control was carried out, without composition according to the invention.
Résultats Results
Les résultats sont représentés sur la figure 1 . La significativité des différences observées entre les différentes conditions testées est mesurée par le test de Student, qui est dit significatif si sa valeur est inférieure à 0,05 (* p<0,05 ; ** p<0,01 ; *** p<0,001) ; ns= non significatif. Le mélange d’hexapeptide Acetyl Hexapeptide-8, de tétrapeptide Palmitoyl Tetrapeptide-7, et de l’extrait aqueux de bois de rose permet de stimuler KI67, marqueur de prolifération cellulaire, au niveau de l’épiderme. Ce résultat démontre que cette association stimule la prolifération des couches basales de l’épiderme et donc favorise son renouvellement. The results are shown in Figure 1. The significance of the differences observed between the different conditions tested is measured by the Student test, which is said to be significant if its value is less than 0.05 (* p<0.05; ** p<0.01; *** p<0.001); ns= not significant. The mixture of hexapeptide Acetyl Hexapeptide-8, tetrapeptide Palmitoyl Tetrapeptide-7, and aqueous extract of rosewood helps stimulate KI67, a marker of cell proliferation, at the level of the epidermis. This result demonstrates that this association stimulates the proliferation of the basal layers of the epidermis and therefore promotes its renewal.
Exemple 3 : Effets des actifs et combinaisons selon l’invention sur l’épaisseur du derme Example 3: Effects of active ingredients and combinations according to the invention on the thickness of the dermis
L’effet des actifs seuls ou en combinaisons selon l’invention, sur l’épaisseur du derme des échantillons de peau reconstruite par bioimpression a été analysé. Chaque test a été effectué en triplicate. The effect of the active ingredients alone or in combinations according to the invention on the thickness of the dermis of skin samples reconstructed by bioprinting was analyzed. Each test was performed in triplicate.
Les échantillons de peau reconstruite par bioimpression ont été testés respectivement avec les actifs et combinaisons suivantes (dissous dans les milieux de culture): l’extrait aqueux de bois de rose seul (concentration finale 0,07%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,62pg/ml) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,19%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,62pg/ml) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,19%) + l’extrait aqueux de bois de rose (concentration finale 0,025%). The skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations (dissolved in the culture media): the aqueous extract of rosewood alone (final concentration 0.07%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.62pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.19%) + aqueous extract of rosewood (final concentration 0.025%).
Un témoin négatif non traité a été effectué, sans composition selon l’invention. An untreated negative control was carried out, without composition according to the invention.
Résultats Results
Les résultats sont représentés sur la figure 2 La significativité des différences observées entre les différentes conditions testées est mesurée par le test de Student, qui est dit significatif si sa valeur est inférieure à 0,05 (* p<0,05 ; ** p<0,01 ; *** p<0,001) ; ns= non significatif. Ces résultats démontrent que le mélange contenant l’hexapeptide Acetyl Hexapeptide-8, le tétrapeptide Palmitoyl Tetrapeptide-7, et l’extrait aqueux de bois de rose augmente l’épaisseur de l’épiderme. Cette association est donc très intéressante pour combattre l’amincissement de l’épiderme observé avec l’âge. The results are represented in Figure 2 The significance of the differences observed between the different conditions tested is measured by the Student test, which is said to be significant if its value is less than 0.05 (* p<0.05; ** p <0.01; ***p<0.001); ns= not significant. These results demonstrate that the mixture containing the hexapeptide Acetyl Hexapeptide-8, the tetrapeptide Palmitoyl Tetrapeptide-7, and the aqueous extract of rosewood increases the thickness of the epidermis. This association is therefore very interesting to combat the thinning of the epidermis observed with age.
L’ensemble de ces résultats montre que l’utilisation combinée d’un extrait aqueux de bois de rose, d’un acetyl hexapeptide-8, d’un palmitoyl tetrapeptide-8 et éventuellement en outre d’une fraction bioactive isolée de pétales de roses, permet d’obtenir un effet synergique antiâge. Exemple 4 : Effets des actifs seuls ou en combinaisons selon l’invention sur le Collagène VII All of these results show that the combined use of an aqueous extract of rosewood, an acetyl hexapeptide-8, a palmitoyl tetrapeptide-8 and possibly also a bioactive fraction isolated from petals of roses, allows you to obtain a synergistic anti-aging effect. Example 4: Effects of active ingredients alone or in combinations according to the invention on Collagen VII
L’effet des actifs seuls ou en combinaisons selon l’invention, sur la production de Collagène VII par les échantillons de peau reconstruite par bioimpression, a été analysé. Chaque test a été effectué en triplicate. The effect of the active ingredients alone or in combinations according to the invention, on the production of Collagen VII by skin samples reconstructed by bioprinting, was analyzed. Each test was performed in triplicate.
Les échantillons de peau reconstruite par bioimpression ont été testés respectivement avec les actifs et combinaisons suivantes selon l’invention (dissous dans les milieux de culture) : la fraction bioactive isolée de roses fraiches autrement nommée ZETA fraction, seule (concentration finale 0,05%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,21 g/ml) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,06%) + l’extrait aqueux de bois de rose (concentration finale 0,005%) ; un mélange d’hexapeptide Acetyl Hexapeptide-8 (concentration finale 0,10pg/ml) et de tétrapeptide Palmitoyl Tetrapeptide-7 (concentration finale 0,003%) + l’extrait aqueux de bois de rose (concentration finale 0,004%) + ZETA fraction (concentration finale 0,003%). The skin samples reconstructed by bioprinting were tested respectively with the following active ingredients and combinations according to the invention (dissolved in the culture media): the bioactive fraction isolated from fresh roses, otherwise called ZETA fraction, alone (final concentration 0.05% ) ; a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.21 g/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.06%) + aqueous extract of rosewood (final concentration 0.005%); a mixture of hexapeptide Acetyl Hexapeptide-8 (final concentration 0.10pg/ml) and tetrapeptide Palmitoyl Tetrapeptide-7 (final concentration 0.003%) + aqueous extract of rosewood (final concentration 0.004%) + ZETA fraction ( final concentration 0.003%).
Un témoin négatif non traité a été effectué, sans actifs ni combinaisons d’actifs selon l’invention. An untreated negative control was carried out, without active ingredients or combinations of active ingredients according to the invention.
Résultats Results
Les résultats sont représentés sur la figure 3. La significativité des différences observées entre les différentes conditions testées est mesurée par le test de Student, qui est dit significatif si sa valeur est inférieure à 0,05 (* p<0,05 ; ** p<0,01 ; *** p<0,001) ; ns= non significatif. Ces résultats montrent que le mélange contenant l’hexapeptide Acetyl Hexapeptide-8, le tétrapeptide Palmitoyl Tetrapeptide-7, l’extrait aqueux de bois de rose et la ZETA fraction stimule de façon très significative et de façon synergique l’expression du collagène de type VII, essentiel pour la bonne structure de la peau. En effet, le collagène VII est une composante essentielle de la matrice extra cellulaire, et donne à la peau sa fermeté. Cette association est donc très avantageuse pour empêcher la perte d’expression du collagène VII observé avec le vieillissement, et donc combattre les signes de l’âge. The results are represented in Figure 3. The significance of the differences observed between the different conditions tested is measured by the Student test, which is said to be significant if its value is less than 0.05 (* p<0.05; ** p<0.01; *** p<0.001); ns= not significant. These results show that the mixture containing the hexapeptide Acetyl Hexapeptide-8, the tetrapeptide Palmitoyl Tetrapeptide-7, the aqueous extract of rosewood and the ZETA fraction very significantly and synergistically stimulates the expression of type collagen. VII, essential for the good structure of the skin. Indeed, collagen VII is an essential component of the extra cellular matrix, and gives the skin its firmness. This association is therefore very advantageous for preventing the loss of expression of collagen VII observed with aging, and therefore combating the signs of aging.
Exemple 5 : Formulations cosmétiques selon l’invention 5.1 Composition sous la forme d’une émulsion Example 5: Cosmetic formulations according to the invention 5.1 Composition in the form of an emulsion
Eau déminéralisée qsp 100,0% Demineralized water qsp 100.0%
Glycols 20,0% Glycols 20.0%
Conservateurs 0,6% Preservatives 0.6%
Chélatant 0,04% Chelating 0.04%
Carbomer (Carbopol® 981 ) 0,3% Carbomer (Carbopol® 981) 0.3%
Sodium polyacrylate (Covacryl® MV60) 0,2% Sodium polyacrylate (Covacryl® MV60) 0.2%
Sodium Hydroxide 0,15% Sodium Hydroxide 0.15%
Extrait aqueux de bois de rose * 1% Aqueous extract of rosewood * 1%
Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprenant 0.05% de peptide) 3% (soit 0,0015% peptide)Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)
Palmitoyl tetrapeptide-7 (RIGIN® comprenant Palmitoyl tetrapeptide-7 (RIGIN® comprising
0.05% de peptide) 4% (soit 0,002% peptide)0.05% peptide) 4% (i.e. 0.002% peptide)
Huile végétale, esters 16% Vegetable oil, esters 16%
Anti-oxydant 0,2% Anti-oxidant 0.2%
Concentré de parfum 0,4% Perfume concentrate 0.4%
Steareth-2 0,8% Steareth-2 0.8%
Steareth-21 1 ,5% Steareth-21 1.5%
* tel que décrit dans l’exemple 1 * as described in example 1
Appliquée sur la peau, en particulier la peau du visage, cette composition confère tonicité, élasticité et fermeté, les traits sont réhaussés, le visage est regalbé. Applied to the skin, in particular the skin of the face, this composition gives tone, elasticity and firmness, the features are enhanced, the face is reshaped.
5.2 : Composition sous la forme d’une crème riche pour le visage Eau déminéralisée qsp 100,0% 5.2: Composition in the form of a rich facial cream Demineralized water qsp 100.0%
Glycols 13,0% Glycols 13.0%
Carbomer (Carbopol® 981 ) 0,3% Carbomer (Carbopol® 981) 0.3%
Sodium polyacrylate (Covacryl® MV60) 0,2% Sodium polyacrylate (Covacryl® MV60) 0.2%
Sodium Hydroxide 0,15% Sodium Hydroxide 0.15%
Extrait aqueux de bois de rose * 0,5% Aqueous extract of rosewood * 0.5%
ZETA fraction ** 0,3% ZETA fraction ** 0.3%
Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprenant 0.05% de peptide) 2% (soit 0,001% peptide) Palmitoyl tetrapeptide-7 (RIGIN® comprenant Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 2% (i.e. 0.001% peptide) Palmitoyl tetrapeptide-7 (RIGIN® comprising
0.05% de peptide) 3% (soit 0,0015% peptide)0.05% peptide) 3% (i.e. 0.0015% peptide)
Huile végétale, esters 16% Vegetable oil, esters 16%
Triglycérides d’acides gras 4% Triglycerides of fatty acids 4%
Beurre de karité 1 % Shea butter 1%
Antioxydantt 0,2% Antioxidant 0.2%
Concentré de parfum 0,4% Perfume concentrate 0.4%
Steareth-2 0,8% Steareth-2 0.8%
Steareth-21 1 ,5% Steareth-21 1.5%
Conservateurs qs Conservatives qs
* et ** tel que décrit dans l’exemple 1 * and ** as described in Example 1
Appliquée sur la peau du visage, cette composition confère tonicité, élasticité et fermeté, les traits sont réhaussés, le visage est regalbé. Applied to the skin of the face, this composition gives tone, elasticity and firmness, the features are enhanced, the face is reshaped.
5.3 : Composition sous la forme d’un gel-sérum pour le contour des yeux 5.3: Composition in the form of a gel-serum for the eye contour
Eau purifiée Qsp 100.00% Purified water Qsp 100.00%
Glycols 13,0% Glycols 13.0%
Conservateurs 0,60% Preservatives 0.60%
Carbomer 0,80% Carbomer 0.80%
Glyceryl stearate citrate 0,70% Glyceryl stearate citrate 0.70%
Lécithine et sodium acrylates copolymer (Lecigel PCR négatif) 1 ,20% Lecithin and sodium acrylates copolymer (Lecigel PCR negative) 1.20%
Isostearate isostearyle 9,0% Isostearyl isostearate 9.0%
Bis-diglyceryl polyacyladipate-2 (Softisan 649 MB) 1 ,0% Bis-diglyceryl polyacyladipate-2 (Softisan 649 MB) 1.0%
Silice 2,0% Silica 2.0%
Nacres 1 ,0% Mother-of-pearl 1.0%
Extrait de Centella asiatica 0,5% Centella asiatica extract 0.5%
Extrait de marron d’inde 0,1 % Horse chestnut extract 0.1%
Extrait aqueux de bois de rose * 0,5% Aqueous extract of rosewood * 0.5%
Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprenant 0.05% de peptide) 3% (soit 0,0015% peptide)Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)
Palmitoyl tetrapeptide-7 (RIGIN® comprenant 0.05% de peptide) 4% (soit 0,002% peptide)Palmitoyl tetrapeptide-7 (RIGIN® comprising 0.05% peptide) 4% (i.e. 0.002% peptide)
ZETA Fraction ** 0,3% Tocopheryl acetate 0,10%ZETA Fraction ** 0.3% Tocopheryl acetate 0.10%
Parfum de rose 0,20% Rose scent 0.20%
* et ** tels que décrits dans l’exemple 1 Après application sur le contour des yeux de ce gel-sérum contribue à défroisser rides et ridules, la zone du regard est raffermie. La jeunesse du regard se reconstitue au fil des applications. * and ** as described in example 1 After application to the eye contour, this gel-serum helps to smooth away wrinkles and fine lines, the eye area is firmed. The youthfulness of the eyes is restored over the course of applications.

Claims

REVENDICATIONS
1. Composition cosmétique comprenant, dans un milieu physiologiquement acceptable :1. Cosmetic composition comprising, in a physiologically acceptable environment:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, et (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8, and
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7. (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7.
2. Composition cosmétique selon la revendication 1 , caractérisée en ce que l’extrait aqueux de bois de rose est obtenu par un procédé d’extraction enzymatique comprenant les étapes : a) addition d'eau au bois de rose préalablement broyé présentant une taille de particule appropriée, b) addition d'un mélange enzymatique contenant au moins une cellulase, au moins une hémicellulase et au moins une pectinase, et avantageusement en outre au moins une protéase, c) incubation sous agitation du bois de rose préalablement broyé et du mélange enzymatique pour libérer dans le milieu réactionnel des huiles, des protéines et des sucres fermentescibles, pendant une durée dépendant des rendements recherchés, d) séparation du milieu réactionnel pour obtenir de l'huile libre, une phase aqueuse contenant des protéines et des sucres fermentescibles, et une phase solide, e) séparation de la phase aqueuse et ajout éventuel de conservateurs. 2. Cosmetic composition according to claim 1, characterized in that the aqueous extract of rosewood is obtained by an enzymatic extraction process comprising the steps: a) addition of water to the previously crushed rosewood having a size of appropriate particle, b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously also at least one protease, c) incubation with stirring of the previously ground rosewood and the mixture enzymatic to release fermentable oils, proteins and sugars into the reaction medium, for a period depending on the desired yields, d) separation of the reaction medium to obtain free oil, an aqueous phase containing fermentable proteins and sugars, and a solid phase, e) separation of the aqueous phase and possible addition of preservatives.
3. Composition cosmétique selon la revendication 1 ou la revendication 2, caractérisée en ce qu’elle comprend en outre un extrait de roses sous la forme d’une fraction bioactive isolée obtenue à partir de roses fraîches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. 3. Cosmetic composition according to claim 1 or claim 2, characterized in that it further comprises a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
4. Composition cosmétique selon la revendication 1 ou 2, caractérisée en ce que : (i) L’extrait aqueux de bois de rose de la variété Evanrat est présent en une teneur allant de 0,1 % à 1 %, en particulier de 0,2% à 0,5% en poids par rapport au poids total de la composition, 4. Cosmetic composition according to claim 1 or 2, characterized in that: (i) The aqueous extract of rosewood of the Evanrat variety is present in a content ranging from 0.1% to 1%, in particular from 0.2% to 0.5% by weight relative to the total weight of the composition,
(ii) L’hexapeptide de nom INCI : Acetyl Hexapeptide-8 est présent en une teneur allant de 0,05% à 0,25% en poids par rapport au poids total de la composition, et (ii) The hexapeptide with INCI name: Acetyl Hexapeptide-8 is present in a content ranging from 0.05% to 0.25% by weight relative to the total weight of the composition, and
(iii) Le tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7 est présent en une teneur allant de 0,1 % à 0,25% en poids par rapport au poids total de la composition. (iii) The tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, is present in a content ranging from 0.1% to 0.25% by weight relative to the total weight of the composition.
5. Composition cosmétique selon la revendication 3, caractérisée en ce que : 5. Cosmetic composition according to claim 3, characterized in that:
(i) L’extrait aqueux de bois de rose de la variété Evanrat est présent en une teneur allant de 0,1 % à 1 %, en particulier de 0,2% à 0,5% en poids par rapport au poids total de la composition, (i) The aqueous extract of rosewood of the Evanrat variety is present in a content ranging from 0.1% to 1%, in particular from 0.2% to 0.5% by weight relative to the total weight of the composition,
(ii) L’hexapeptide de nom INCI : Acetyl Hexapeptide-8 est présent en une teneur allant de 0,05% à 0,25% en poids par rapport au poids total de la composition, (ii) The INCI name hexapeptide: Acetyl Hexapeptide-8 is present in a content ranging from 0.05% to 0.25% by weight relative to the total weight of the composition,
(iii) Le tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7 est présent en une teneur allant de 0,1 % à 0,25% en poids par rapport au poids total de la composition, et (iii) The tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, is present in a content ranging from 0.1% to 0.25% by weight relative to the total weight of the composition, and
(iv) La fraction bioactive isolée de roses fraiches de la variété Evanrat, est présente en une teneur allant de 0,1 à 1 % et en particulier de 0,2 à 0,5% en poids par rapport au poids total de la composition. (iv) The bioactive fraction isolated from fresh roses of the Evanrat variety is present in a content ranging from 0.1 to 1% and in particular from 0.2 to 0.5% by weight relative to the total weight of the composition .
6. Composition cosmétique selon l’une quelconque des revendications 1 à 5, caractérisée en ce qu’il s’agit d’une composition de soin et/ou de maquillage de la peau et/ou des lèvres, en particulier d’une composition de soin de la peau. 6. Cosmetic composition according to any one of claims 1 to 5, characterized in that it is a care and/or makeup composition for the skin and/or lips, in particular a composition skin care.
7. Procédé cosmétique non thérapeutique de soin et/ou de maquillage de la peau et/ou des lèvres comprenant l’application sur ladite peau et/ou lesdites lèvres, en particulier la peau, d’une composition telle que définie dans l’une quelconque des revendications 7. Non-therapeutic cosmetic process for caring for and/or making up the skin and/or lips comprising the application to said skin and/or said lips, in particular the skin, of a composition as defined in one any of the claims
8. Procédé cosmétique selon la revendication 7, caractérisé en ce que l’application de la composition sur la peau et/ou les lèvres permet de prévenir et/ou diminuer les signes du vieillissement cutané. 8. Cosmetic process according to claim 7, characterized in that the application of the composition to the skin and/or lips makes it possible to prevent and/or reduce the signs of skin aging.
9. Association d’actifs cosmétiques comprenant : 9. Combination of cosmetic active ingredients including:
(i) Un extrait aqueux de bois de rose de la variété Evanrat, de préférence de bois de rose d’été, obtenu par extraction enzymatique au moyen d’un solvant polaire cosmétiquement acceptable, (i) An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent,
(ii) Un hexapeptide de nom INCI : Acetyl Hexapeptide-8, (ii) A hexapeptide with the INCI name: Acetyl Hexapeptide-8,
(iii) Un tétrapeptide Pal-GQPR-NH2 ou Pal-GQPR-OH, en particulier le tétrapeptide de nom INCI : Palmitoyl Tetrapeptide-7, et (iii) A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7, and
(iv) Eventuellement en outre une fraction bioactive isolée obtenue à partir de roses fraiches de la variété Evanrat et comprenant au moins 500 pg/pL d’au moins un sucre monosaccharide choisi parmi le fructose, glucose et le saccharose, et/ou au moins 500 mg/Kg d’au moins un minéral choisi parmi le potassium, le calcium et le sodium. (iv) Optionally also an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and comprising at least 500 pg/pL of at least one monosaccharide sugar chosen from fructose, glucose and sucrose, and/or at least 500 mg/Kg of at least one mineral chosen from potassium, calcium and sodium.
10. Utilisation cosmétique non thérapeutique de l’association d’actifs cosmétiques telle que définie dans la revendication 9, comme association active pour prévenir et/ou diminuer les signes du vieillissement cutané, en particulier la perte de densité et/ou de fermeté de la peau, la diminution de l’épaisseur de l’épiderme, la diminution de la différenciation et/ou prolifération des cellules épidermiques, et/ou l’apparition de rides et/ou ridules. 10. Non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined in claim 9, as an active association for preventing and/or reducing the signs of skin aging, in particular the loss of density and/or firmness of the skin. skin, the reduction in the thickness of the epidermis, the reduction in the differentiation and/or proliferation of epidermal cells, and/or the appearance of wrinkles and/or fine lines.
PCT/FR2023/050797 2022-06-07 2023-06-06 Anti-ageing composition with rose extracts and peptides WO2023237833A1 (en)

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