Anti-aging composition with rose extracts and peptides

Abstract

The present invention relates to a cosmetic composition comprising, in a physiologically acceptable medium: An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, A hexapeptide with INCI name: Acetyl Hexapeptide-8, A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, andOptionally furthermore a rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety.

Description

Title of the invention: Anti-aging composition with rose extracts and peptides FIELD OF THE INVENTION

[0001] 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. STATE OF THE ART

[0002] The skin is the body's first barrier of protection against the environment. It is subjected daily to the effects of factors of exogenous origin (e.g. UV radiation, temperature variations, atmospheric pollution, cigarette smoke, etc.) and / or endogenous origin (e.g. hormones, etc.).

[0003] Aging causes changes in the skin of the body and / or face, including the lips, areas particularly subject to the harmful effects of the environment, such as wrinkles and fine lines, skin that has lost its firmness, suppleness and elasticity, a reduction in the thickness of the epidermis linked in particular to a slowdown in the renewal of skin cells.

[0004] A large number of anti-aging active ingredients are known from the prior art, such as peptides, vitamins, antioxidants and plant extracts. Examples include the peptides Acetyl Hexapeptide-8 from Lipotec and Palmitoyl Tetrapeptide-7 from Sederma.

[0005] The use of certain rose extracts in anti-aging is also known, such as the aqueous extract of rosewood from the Evanrat variety described in patent application FR3090380.

[0006] However, there is a constant need to find new compounds or new combinations of active ingredients having improved properties in the prevention and / or aging of the skin.

[0007] Unexpectedly, the Applicant has demonstrated a synergistic in vitro anti-aging effect between the peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7) and an aqueous extract of rosewood of the Evanrat variety. It has also demonstrated an anti-aging synergy between the peptides (Acetyl Hexapeptide-8 and Palmitoyl tetrapeptide-7), an aqueous extract of 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. Statement of the invention

[0008] The present invention therefore relates in particular to a cosmetic composition comprising, in a physiologically acceptable environment:

[0009] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with the INCI name: Acetyl Hexapeptide-8, and iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7.

[0010] 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 as defined in the present invention.

[0011] The invention also relates to a combination of cosmetic active ingredients comprising:

[0012] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with INCI name: Acetyl Hexapeptide-8, iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and iv. Optionally, in addition, 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.

[0013] 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 combination for preventing and / or reducing 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.

[0014] The invention also relates to a cosmetic assembly comprising:

[0015] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with INCI name: Acetyl Hexapeptide-8, iii. A tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and iv. Optionally, in addition, 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,

[0016] 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.

[0017] The invention finally relates to a cosmetic process comprising the successive application, to the skin and / or the lips, of the compositions of the cosmetic assembly as defined in the present invention. DESCRIPTION OF FIGURES

[0018] [Fig.l] 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.

[0019] [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.

[0020] [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. DETAILED DESCRIPTION OF THE INVENTION

[0021] Thus, the present invention relates in particular to a cosmetic composition comprising, in a physiologically acceptable medium:

[0022] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with the INCI name: Acetyl Hexapeptide-8, and iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with the INCI name: Palmitoyl Tetrapeptide-7.

[0023] 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 anciennes André Eve SAS" and protected by Plant Variety Certificate under No. 20110345 with the species name Rosa L. and the variety name EVANRAT. This bush rose bush belongs to the group of modern hybrids, which, from May to October, is permanently covered in roses, thus showing an excellent remontant character. Rosewood extract and preparation process

[0024] The aqueous extract of rosewood of the Evanrat variety or rose 'Jardin de Granville®', 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.

[0025] 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 a process of eco-extraction of rosewood or rosewood in the description.

[0026] The expression "polar solvent" means that the solvent has a Polarity Index value that 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 character of a molecule. For the polarity indices of solvents, refer to the article by LR SNYDER: Classification of the solvent properties of common liquids; Journal of Chromatography, 92 (1974), 223-230.

[0027] The preferred polar solvents are those consisting of a compound comprising at least one polar covalent bond of OH type.

[0028] As cosmetically acceptable polar solvent, a solvent or a mixture of solvents is 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.

[0029] According to a particular embodiment, 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.

[0030] Plant material

[0031] There are two types of rosewood depending on the time of year they are harvested:

[0032] - Winter woods, generally harvested between the months of November and April

[0033] - Summer woods, generally harvested between the months of May and October.

[0034] According to a particular and preferred mode, summer woods will be used. In the description, we will speak indifferently of rosewood or summer rosewood.

[0035] The rosewoods according to the invention are obtained from freshly cut whole roses and are isolated from the rest of the plant by manual or mechanical separation.

[0036] In a particular embodiment, the rosewoods are directly used for extraction.

[0037] In another embodiment, the rosewoods are previously dried before extraction.

[0038] According to a particular and preferred embodiment, the rosewood is ground prior to the extraction step.

[0039] Rosewoods include monosaccharide sugars (fructose, glucose), and polysaccharides in particular disaccharides, polyphenols (catechin), acids amino acids (mainly aspartic acid, tyrosine and arginine), minerals (ash, sodium, potassium, calcium), flavonoids (mostly glycosylated) and tannins.

[0040] In particular, calcium improves epidermal differentiation and strengthens skin structure, while potassium amplifies skin hydration and boosts energy assimilation. Phytosugars (fructose, glucose) are intended to fill skin cells with energy.

[0041] The aqueous extract of rosewood 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 implementing the extraction of natural compounds of rosewood in the presence of aqueous (polar) solvents 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.

[0042] According to a particular embodiment, the aqueous extract of rosewood according to the invention further comprises in 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 extract of rose according to the invention. In this embodiment, the original plant material comprises rosewoods and rose flowers in a proportion of 0.01% flowers for 99.99% of wood to 1% flowers for 99% of wood.

[0043] Extraction process

[0044] 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.

[0045] According to a particular and preferred embodiment, rosewoods of the Evanrat variety, preferably the Jardin de Granville® rose, preferably previously dried rosewoods, will be used as plant material in the extraction processes adapted to the invention described below.

[0046] The woods can be used fresh, frozen or previously dried using conventional drying tools known to those skilled in the art, such as for example 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 woods may have been dried and / or ground. According to another embodiment, the woods are used wet and then ground.

[0047] According to a particular embodiment, the wood is ground before extraction, for example by means of a mortar, cryogenic grinding, a mixer, a traditional grinder or a centrifuge according to methods known to those skilled in the art.

[0048] The aqueous extract of rosewood according to the invention can be obtained in particular according to enzymatic extraction processes.

[0049] Depending on the molecules that one seeks to extract, the enzymes will be adapted to the walls to be hydrolyzed to access these said molecules. Advantageously, one can in particular use cellulase, hemicellulase, pectinase or protease type enzymes, alone or in combination.

[0050] It will thus be possible to use a cellulase to release the glucose and the cellulose; a hemicellulase to release the mono and oligomers of reduced sugars; a pectinase to release the uronic acids; and advantageously a protease to hydrolyze the structural proteins and improve the lysis of the raw material.

[0051] 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 three types of products from plant matter in a single step, namely an oil (rich in nutrients polyphenols, sterols, vitamin E, etc.), an aqueous extract and a cake (solid phase).

[0052] 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:

[0053] a) adding water to the previously ground rosewood having a suitable particle size,

[0054] b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously in addition at least one protease,

[0055] c) incubation with stirring of the previously ground rosewood and the enzymatic mixture to release into the reaction medium oils, proteins and fermentable sugars, for a duration depending on the desired yields,

[0056] d) separation of the reaction medium to obtain free oil, an aqueous phase containing proteins and fermentable sugars, and a solid phase, and

[0057] e) separation of the aqueous phase and possible addition of preservatives.

[0058] Said rosewood extract used according to the invention being obtained after the separation step (e).

[0059] According to a particular embodiment, 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 the particles must have a size close to 50 μm, preferably close to 10 μm.

[0060] Preferably, the mass of water added to the plant material is equal to 1 to 2 times the mass of said plant matter and does not exceed this quantity.

[0061] In particular in step b), the ratio between the activity of the pectinase and the activity of the cellulase 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 activity of the pectinase and the activity of the hemicellulase being at least 7.103; preferably between 1.102 and 0.5, and more preferably between 1.102 and 2.102.

[0062] More particularly, the enzymatic mixture used contains at least one cellulase, at least one hemicellulase, at least one pectinase, and advantageously in addition at least one protease. According to a particular embodiment, the cellulase(s), hemi-cellulase(s) and pectinase(s) represent 75% of the enzymatic mixture and the protease(s) represent 25% of said enzymatic mixture. According to a particular embodiment, 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.

[0063] In a preferred embodiment the enzymatic mixture used contains 25% cellulases, 25% hemicellulases, 25% pectinases and 25% proteases.

[0064] 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.

[0065] 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.

[0066] The hydrolysis reaction is then stopped by deactivating the enzymes by heating, preferably between 80°C and 105°C for, for example, 5 to 20 minutes.

[0067] Then the reaction medium is separated, according to step d), using all suitable known separation techniques, such as centrifugation or decantation.

[0068] According to a particular embodiment, the mixture is pre-filtered on a 0.8mm then 0.5mm sieve in order to remove the largest part of the solid phase, then a sterilizing filtration is advantageously carried out on clarifying plates with a pore size of 0.2pm, at a pressure of 2 Bars.

[0069] The aqueous phase obtained after sterilizing filtration is stabilized with preservatives (citric acid, potassium sorbate, sodium benzoate).

[0070] 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:

[0071] a) adding water to the previously ground rosewood having a suitable particle size,

[0072] b) addition of an enzymatic mixture containing at least one cellulase, at least one hemicellulase and at least one pectinase, and advantageously in addition at least one protease,

[0073] c) incubation with stirring of the previously ground rosewood and the enzymatic mixture to release into the reaction medium oils, proteins and fermentable sugars, for a duration depending on the desired yields,

[0074] d) separation of the reaction medium to obtain free oil, an aqueous phase containing proteins and fermentable sugars, and a solid phase,

[0075] e) separation of the aqueous phase and possible addition of preservatives.

[0076] 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).

[0077] The aqueous extract of rosewood used according to the invention is also characterized by the fact that it is not a fermented extract.

[0078] According to a particular embodiment, 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 extract of rosewood according to the invention is called in the INCI name water (and) rose extract (and) citric acid (and) potassium sorbate (and) sodium benzoate.

[0079] 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

[0080] The compositions of the invention also comprise an acetyl hexapeptide and a palmitoyl tetrapeptide, in particular the following peptides:

[0081] 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.

[0082] According to a particular embodiment, the commercial product ARGIRELINE® Amplified peptide containing Tacetyl 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.

[0083] 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.

[0084] Palmitoyl tetrapeptide-7 which is a synthetic peptide of sequence Palmitoyl-Gly-Gln-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with INCI name: Glycerin, Lactic Acid and Palmitoyl Te-trapeptide-7.

[0085] According to a particular embodiment, the commercial product RIGIN® containing palmitoyl te-trapeptide-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.

[0086] 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.

[0087] According to a particular and preferred embodiment, the composition of the invention further comprises a bioactive fraction isolated from roses as described below.

[0088] Thus, the composition of the invention will generally comprise:

[0089] 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. The hexapeptide with 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, and iii. The tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7 in a content ranging from 0.1% to 0.25% by weight (of peptide) relative to the total weight of the composition, and 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.

[0090] Bioactive fraction isolated from roses and process for its preparation

[0091] By "bioactive fraction" or "isolated bioactive fraction of roses" or "Rose De Granville® Serum", "Zêta" or "Zêta fraction", is meant 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 rose petals as described in application FR3066388.

[0092] Plant material

[0093] 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.

[0094] According to a particular embodiment, summer roses will be used to prepare the bioactive fraction isolated from roses, in particular summer rose petals. According to another embodiment, winter roses will preferably be used, in particular winter rose petals.

[0095] 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 carotene.

[0096] 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.

[0097] By mineral-rich is meant a rose extract in the form of an isolated bioactive fraction, wherein said extract comprises at least 500 mg / Kg of at least one mineral chosen from potassium, calcium and sodium.

[0098] 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.

[0099] Advantageously, calcium improves epidermal differentiation and strengthens the skin's structure, while potassium amplifies skin hydration and boosts energy assimilation. Phyto-sugars (fructose, glucose, sucrose) help to fill skin cells with energy.

[0100] Extraction process

[0101] Fresh roses of the Evanrat variety or 'Jardin de Granville®' roses are used as plant material. According to a preferred method, these are fresh petals of roses of the Evanrat variety or 'Jardin de Granville®' roses.

[0102] Advantageously, said method does not require the addition of any solvent or exogenous liquid.

[0103] The process implemented to obtain the isolated bioactive fraction used according to the invention comprises the main steps of:

[0104] 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 in fibers (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.

[0105] 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" within the meaning of the invention.

[0106] By "cleaning" is meant 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 may be carried out by low-pressure rinsing with potable water under conditions where the runoff wash would not substantially contain plant pigments. Excess wash water is then removed from the washed plants.

[0107] By "maceration" is meant the act of transforming the fresh Granville® roses, and preferably the fresh petals, into smaller particles to break their integrity and subsequently facilitate the expulsion of the liquid intracellular colloidal dispersion (ICD). Examples of suitable maceration instruments include, but are not limited to, devices such as a crusher, grinder, or mill (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.

[0108] By "pressing" is meant the separation of liquid material from fresh Granville® roses, and preferably fresh petals, by the 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 a type suitable for a press.

[0109] By “fiber-enriched material” or “MEF” or “FEM” (Fiber Enriched Material), means a solid and / or semi-solid fraction enriched with fresh Granville® rose fibers, preferably fresh petals, from which the liquid intracellular colloidal dispersion (ICD) has been removed by pressing.

[0110] By "intracellular colloidal dispersion" or "ICD" or "DCI" 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 consist primarily of plant cell organelles, organelle fragments, and fiber-enriched residual material. The aqueous medium consists primarily of cytosols and vacuoles.

[0111] By "separation" is meant the separation of solid and / or semi-solid particles, and non-aqueous liquid droplets 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 preclude batch processing. “Separation 1” and “Separation 2” refer to the respective process steps, carried out with respective parameters.

[0112] By "supernatant" is meant an aqueous material from which particles have been separated. "Supernatant A" and "Supernatant B" mean the supernatants resulting from the respective separation steps of the process.

[0113] By "precipitate" is meant the particles from which an aqueous material has been separated. "Fraction B" and "Fraction C" mean the precipitates resulting from the respective separation steps of the process.

[0114] The terms "fresh Granville® Rose serum fraction" and "fresh Granville® Rose petal serum fraction" refer to compositions produced by the process as shown above without added preservatives and / or stabilizers to protect the ingredient composition from environmental factors such as temperature, atmosphere (e.g., oxygen), light, and microorganisms.

[0115] By "preservatives and / or stabilizers" is meant substances which, when added to a "fraction of serum from fresh Granville® roses", preferably a "fraction of serum from fresh Granville® roses petals", protect it against environmental factors such as temperature, atmosphere (e.g. oxygen), light and microorganisms. Suitable substances part Particulars may include, without limitation, a preservative, a stabilizer and / or a mixture thereof.

[0116] The term "Granville® Rose Serum" or "Fresh Granville® Rose Extract" as used herein means a combination of a fresh Granville® Rose Serum fraction and preservatives and / or stabilizers.

[0117] The term "Granville® rose petal serum" or "Granville® fresh rose petal extract" as used herein means a combination of a Granville® fresh rose petal serum fraction and preservatives and / or stabilizers. In a preferred embodiment, the extract used according to the invention is obtained by carrying out the extraction method disclosed in EP2919757, JP 6130924, CN ZL201380057567.6, EP2491939B1, CN1929851B, US Patent Nos. 8,734,861 and 7,473,435; US Patent Application No. 16 / 078925.

[0118] Preferably, rose flowers (Roses de Granville®) and 4 to 5 cm of stem are harvested in a manner that avoids chopping or crushing the collected biomass to avoid disruption of the cellular structure of the flowers. The viability of the collected plants can be tested using an OS5p multi-mode chlorophyll fluorometer (Opti-Sciences Inc, Hudson, NH, USA).

[0119] According to a particular embodiment, the fresh living 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 immediately at negative temperatures between -20°C and -80°C, such as for example -20°C, -40°C or -60°C. The flowers can thus be stored for long periods, such as for example several months, or several years, before being used for the purposes of the extraction process.

[0120] According to a preferred embodiment, the fresh live flowers, including the petals, pistil and stamen, are removed from the stem, including the sepal and receptacle, and packed in 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 harvesting is complete.

[0121] Step a

[0122] Once harvesting is complete, the roses, preferably the 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 may then undergo maceration, pressing, and separation by mechanical, roller, hydraulic, or juice extractor pressing to extract the contents of the liquid intercellular colloidal dispersion (ICD) from the fiber-enriched material ("Fraction A").

[0123] At the end of these steps, the yield of fraction A is between 30% and 65%, preferably between 35% and 60% and more preferably between 40% and 55% (weight / weight).

[0124] The DCI typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and more preferably from 6% to 12% dry matter.

[0125] At this stage, the DCI can be frozen for storage. Typically, it is frozen at -20°C.

[0126] Step b

[0127] When the DCI has been frozen for storage, it must first be gently thawed. Typically, it is placed to thaw at 4°C or in ice.

[0128] “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 decrease in the value of the real component of the low-frequency dielectric constant (e'o) by approximately 20 Farads per meter (F / m) compared to its value before the treatment. This treatment degrades the stability of the DCI by causing the agglomeration and / or aggregation of particles (i.e., organelles, fragments of organelles, residual fibrous material) into assemblies sufficiently large and stable to allow and / or improve mechanical separation.

[0129] Indeed, the resulting intracellular colloidal suspension is considered to be 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 electric double layers. The energy barrier resulting from the repulsive force prevents the particles of the dispersed phase from approaching each other unless they have enough energy to overcome this barrier, in which case the attractive force will bring them into contact (they will then adhere irreversibly). The DLVO theory describes the interaction and potential energy of the particles as a function of 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 altered conditions in Treatment A allow the repulsive force of the double layer to decrease to the point where there is no longer a potential energy barrier and the particles . can approach and congregate freely.

[0130] Restoring the initial conditions does not restore stability, because the particles have irreversibly agglomerated. They are thus easily removed by mechanical means (Koganov et al., sofw journal 2017).

[0131] 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”.

[0132] Typically, "supernatant A" has a turbidity of less than about 100 NTU.

[0133] “Fraction B” typically comprises from 10% to 30% dry matter, preferably from 13% to 27% dry matter, and more preferably about 15.0% to 25.0% dry matter.

[0134] Step c

[0135] “Treatment B” is carried out by adjusting the pH level in “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.

[0136] 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”.

[0137] “Fraction C” typically comprises from 5% to 25% dry matter, preferably from 8% to 22% dry matter, and more preferably about 10.0% to 20.0% dry matter.

[0138] Step d

[0139] “Treatment C” is carried out by adjusting the pH level in “supernatant B”, in particular by titration with, for example, acid until a pH of less than 4.5 is obtained. Typically, a citric acid solution will be used as the preferred acid.

[0140] 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 “fraction of serum from fresh Granville® roses”, preferably the “fraction of serum from fresh Granville® roses petals” (Non-Preserved Extract) and “Fraction D”.

[0141] The “Granville® fresh rose petal serum fraction” typically comprises from 2% to 20% dry matter, preferably from 4% to 16% dry matter, and more preferably from 6.0% to 10.0% dry matter.

[0142] Step e

[0143] 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 thereof to give the rose extract de Granville® fresh or “Serum Rose De Granville®”, preferably the extract of fresh petals of roses de Granville® or “Serum of petals of Roses De Granville®”.

[0144] 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 may include at least one preservative, at least one stabilizer, at least one antioxidant, or mixtures thereof.

[0145] 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:

[0146] 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 in fibers (fraction A);

[0147] b) destabilization of the DCI with electromagnetic waves then mechanical separation of the intracellular colloidal dispersion (DCI) to obtain the supernatant A and a Membrane Fraction (fraction B);

[0148] c) adjusting the pH level in supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanical separation of supernatant A to obtain supernatant B and fraction C (cytoplasmic fraction);

[0149] d) adjusting the pH level in supernatant B until a pH of less than 4.5 is obtained, then mechanically separating supernatant B to give a “Bioactive Serum Fraction” and a fraction D (precipitate); and

[0150] e) Optionally, mixing the “bioactive serum fraction” with at least one preservative and / or stabilizer.

[0151] According to a particular embodiment, 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 in the INCI name Rosa Hybrid Flower Extract.

[0152] According to another particular embodiment, 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.

[0153] 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. weight relative to the total weight of the composition.

[0154] Thus, in a particular embodiment, the cosmetic composition according to the invention comprises:

[0155] 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. a hexapeptide with 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. a tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7 in a content ranging from 0.1% to 0.25% by weight (of peptide) relative to the total weight of the composition, and 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. Cosmetic and galenic composition

[0156] 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, in particular the skin of the face and / or the neck.

[0157] 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.

[0158] The physiologically acceptable medium generally represents from 1 to 99% by weight, relative to the total weight of said composition.

[0159] 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.

[0160] According to a particular embodiment, the cosmetic composition according to the invention is characterized in that it is a composition for caring for and / or making up the skin and / or the lips, in particular a skin care composition.

[0161] A composition for topical application according to the invention may be, for example, 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.

[0162] 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.

[0163] In a preferred embodiment, said composition used according to the invention is in the form of a cream or a serum.

[0164] The aqueous phase of the composition according to the invention comprises water and optionally a water-soluble solvent.

[0165] According to the invention, the term 'water-soluble solvent' means 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). In particular, mention may be made of:

[0166] - lower C1-C5 monoalcohols such as ethanol, isopropanol and their mixtures;

[0167] - C2-C8 glycols such as ethylene glycol, propylene glycol, 1,3-butylene glycol, dipropylene glycol, and mixtures thereof;

[0168] - C2-C32 polyols such as polyglycerols, polyethylene glycols, and their mixtures,

[0169] and mixtures thereof.

[0170] It may also comprise hydrophilic gelling agents, antioxidants, preservatives and mixtures thereof.

[0171] The cosmetic composition according to the invention may further comprise a fatty (solid fatty substances) or oily phase.

[0172] The term "oily phase" means an oil or a mixture of oils which are miscible with each other, or not. For the purposes of the invention, the term "oil" means a fatty substance, insoluble in water, liquid at 25°C and atmospheric pressure. These oils may be volatile or non-volatile, vegetable, mineral or synthetic.

[0173] An oily phase according to the invention may comprise natural, hydrocarbon, silicone oils, and their mixtures.

[0174] 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 relative to the total weight of said composition.

[0175] 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, pearlescent agents and mixtures thereof.

[0176] 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 rose fresh, 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, pearlescent agents and mixtures thereof. Cosmetic process

[0177] 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 as defined according to the invention.

[0178] By "skin and / or lips" according to the invention, we mean in particular healthy skin and / or lips, that is to say not presenting any troubles or disorders which would be a pathological condition ('unhealthy' subjects, suffering from a pathology).

[0179] According to a particular embodiment, the application of the composition as defined according to the invention to the skin and / or the 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

[0180] The present invention also relates to a combination of cosmetic active ingredients comprising:

[0181] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with INCI name: Acetyl Hexapeptide-8, iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and iv. Optionally, in addition, 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.

[0182] The invention also relates to the non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined above, as an active combination for preventing and / or reducing 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. Cosmetic set and use

[0183] The present invention also relates to a cosmetic assembly comprising:

[0184] i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with INCI name: Acetyl Hexapeptide-8, iii. A tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and iv. Optionally, in addition, 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,

[0185] 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.

[0186] Another object is a cosmetic process comprising the successive application, to the skin and / or the lips, of the compositions of the cosmetic assembly as defined above.

[0187] 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. EXAMPLES Example 1: Materials and methods 1.1 Preparation of cell cultures

[0188] The various tests of the ingredients alone or in combination according to the invention were carried out on samples of skin reconstructed by extrusion bioprinting with cells from a mature donor (= 42 years).

[0189] Cells used

[0190] All of the cells used were extracted by LabSkin Créations from surgical residues of donors who had undergone plastic or reconstructive surgery. The collection of these surgical residues was obtained in accordance with current French regulations and authorized by the Ministry of Research under number DC2020-4346.

[0191] In order to ensure their use in tissue engineering, primary cell banks were created at different passages depending on the cell type considered. The cells were secured from a microbiological point of view and are free of mycoplasmas.

[0192] [Tables 1] Cells of interest Donor characteristics Culture passage 3D thawing Fibroblasts NHDF Adult 42 years P5 P7 Keratinocytes NHEK Adult 42 years PI P3

[0193] Bioprinting and culture of reconstructed skins

[0194] Bioprinting of dermis

[0195] Fibroblasts from aged donors were trypsinized and counted. The cells were suspended in bioink consisting of alginate, gelatin and fibrinogen (LabSkin Créations) at a rate of 250,000 cells per mL of ink. The bioinks were transferred into sterile syringes mounted on 10 gauge cannulas.

[0196] The printed objects were modeled with Slic3r software (GNU Affero General Public License) and Repetier (Hot-World GmbH & Co. KG). The dermis were printed using a 3D printer (LabSkin Créations) to a size of 1.5cm x 1.5cm and comprised two layers of cells extruded into the bioink.

[0197] The constructs were then polymerized for 1 hour in a calcium and thrombin solution (LabSkin Créations) allowing consolidation of the bioink. After three rinses in a saline buffer, the equivalent dermis were cultured in DMEM containing 10% calf serum and 1% antibiotics and antifungals and placed in an incubator (37°C, 5% CO2) for 12 days. The culture medium was renewed every 2 days.

[0198] Epidermization

[0199] 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 calf serum, hydrocortisone and insulin was added to submerge the bioprinted dermo-epidermal constructs.

[0200] Maturation and differentiation

[0201] 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 multi-stratified epidermis.

[0202] Treatments

[0203] 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. Preparation of rosewood extract:

[0204] The rosewoods of the Evanrat variety, in particular the Jardin de Granville® rose variety, possibly with the addition of rose flowers, were reduced to powder in three stages.

[0205] Step 1: Pass through a garden shredder to obtain two-centimeter sections

[0206] Step 2: passing through an industrial grinder

[0207] Solution

[0208] The enzymatic extraction was carried out in a reactor with a maximum capacity of 701. The reactor is stirred by a propeller and is thermostatically controlled by a double jacket.

[0209] 16 kg of rosewood powder with 0.1% rose flowers are dispersed and suspended in 47.5 kg of water.

[0210] In order to deactivate the endogenous enzymes, the mixture was raised to a temperature of 85°C for 1 min and then stabilized at 50°C.

[0211] Lysis of plant walls

[0212] 0.8 Kg of a HEL1PR1 Enzyme Cocktail comprising a mixture of cellulases, 75% hemicellulases and pectinases and 25% proteases were added to the corresponding reaction mixture (5% w / w). The reaction was continued for 4 hours, the temperature was maintained at 50°C.

[0213] Inactivation of enzymes

[0214] The mixture was brought to 85°C for 1 min to inactivate the enzymes, then brought back to 35°C.

[0215] Pre-filtration

[0216] The mixture is pre-filtered on a 0.8mm then 0.5mm sieve in order to remove the largest part of the solid phase, then on a sheet.

[0217] Sterilizing filtration

[0218] Filtration was carried out on clarifying plates (PALL corporation) with a pore size of 0.2 pm, at a pressure of 2 Bars.

[0219] Formulation

[0220] The aqueous phase obtained after sterilizing filtration is stabilized with the following preservatives:

[0221] - Citric acid,

[0222] - Potassium sorbate and

[0223] - Sodium benzoate.

[0224] After filtration, an aqueous extract of rosewood is obtained at 3.7% by weight of dry matter relative to the total weight of the extract.

[0225] Preparation of the active fraction of fresh roses (ZETA fraction):

[0226] Fresh petals of roses of the Evanrat variety or “Jardin de Granville®” rose are used as plant material.

[0227] The bioactive fraction of rose petals used according to the invention is obtained according to the following protocol:

[0228] al) cleaning fresh rose petals by spraying with water at a temperature of about 12°C, for 0.1 to 0.3 minutes at a flow rate of 5 to 6 liters per minute,

[0229] a2) maceration, pressing then mechanical separation of the roses using a press mechanical screw (model CP-6 Vincent Corporation, FL) to extract the contents of the liquid intercellular colloidal dispersion (ICD) from the fiber-enriched material (“Fraction A”);

[0230] bl) destabilization of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz,

[0231] b2) centrifugation of the intracellular colloidal dispersion (ICD) to obtain the supernatant A and a Membrane Fraction (fraction B);

[0232] cl) adjusting the pH level in supernatant A with an alkali (i.e., potassium carbonate), in order to obtain a pH ranging from 6 to 7,

[0233] c2) mechanical separation of supernatant A to obtain supernatant B and the fraction C (cytoplasmic fraction);

[0234] dl) adjusting the pH level in supernatant B with citric acid to obtain a pH lower than 4.5,

[0235] d2) mechanical separation of the supernatant B to give a “serum fraction of fresh petals of Granville® roses” and a fraction D (precipitate); and

[0236] e) mixture of the “fraction of serum from fresh petals of Granville® roses” with potassium sorbate and sodium benzoate.

[0237] 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 material), 91.5% by weight of water, 0.15% by weight of potassium sorbate and 0.3% by weight of sodium benzoate. These contents are understood to be by weight relative to the total weight of the extract. The INCI name of this extract is Rosa Hybrid Flower Extract, Potassium Sorbate, and Sodium Benzoate.

[0238] 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.

[0239] The percentages indicated in the examples correspond to the percentages by weight of commercial product in the culture media or in the cosmetic formulas.

[0240] Palmitoyl tetrapeptide-7 is a synthetic peptide of sequence Palmitoyl-Gly-Gln-Pro-Arg (Palmitoyl-GQPR) from the company SEDERMA marketed under the name RIGIN® with INCI name: Glycerin, Lactic Acis and Palmitoyl Te-trapeptide-7. In the examples, the term “Palmitoyl tetrapeptide-7” means the commercial product RIGIN® comprising 0.05% of peptide. The percentages indicated in the examples correspond to the percentages by weight of commercial product in the culture media or in the cosmetic formulas.

[0241] The concentrations of the active ingredients or combinations were selected based on pre-performed cytotoxicity results.

[0242] The reconstructed skins were treated during dermal reconstruction, from D5 to D12 and epidermal reconstruction from D14 to D18 then from D20 to D26.

[0243] Stopping crops

[0244] 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) and then embedded in paraffin.

[0245] 1.2 Methods for analyzing bioprinted skin samples after tests

[0246] The different treatments (active ingredients alone or in combination) were tested on the samples of skin reconstructed by bioprinting according to example 1. Different analyses were carried out on these samples of skin reconstructed by bioprinting to determine the effect of each treatment on the skin.

[0247] Hematoxylin-Phloxine-Saffron Staining

[0248] 5 μm paraffin sections were made on the different samples of skin reconstructed by bioprinting. After deparaffinization and rehydration, the samples were stained with Hematoxylin, Phloxine and Saffron (HPS) by three successive 5 min baths. After rinsing, the sections were dehydrated before being mounted on a slide with a xylene-based hydrophobic mounting medium.

[0249] Immunofluorescence on paraffin sections

[0250] All labeling was performed on 5 pm thick sections. After heat unmasking, a saturation step in PBS / BSA 5% allowed blocking of nonspecific antigenic sites. The cells were then incubated overnight with a primary antibody specific to the antigen of interest:

[0251] - Anti Ki-67 antibody: 1 / 50th monoclonal, mouse, Dako;

[0252] - Anti-Collagen VII antibody: 1 / 50th, monoclonal, mouse, Santa Cruz.

[0253] The next day, after rinsing steps, the sections were incubated with an Alexa-Fluor 568 anti-mouse secondary antibody diluted to 1 / 1000 (Thermo Fisher Scientific) and the Hoechst nuclear stain (Thermo Fisher Scientific). A negative control without primary antibody was carried out in parallel.

[0254] Image acquisition

[0255] Histological stainings and immunohistological markings were observed with the Axio Observer DI optical microscope system / Axiocam high-resolution camera (Zeiss, Le Pecq, France). Images were acquired using ZEN pro 2012 software and saved in “tif” format (high resolution).

[0256] Image analysis

[0257] Image analysis was performed with ImageJ software (Rasband, WS, ImageJ, US National Institutes of Health, Bethesda, Maryland, USA, http: / / imagej.nih.gov / ij / , 1997-2017). For each parameter, a statistical analysis was performed with a Student test, which is significant if its value is less than 0.05. The results are indicated as: * P < 0.05, ** P < 0.01 and *** P < 0.001.

[0258] Thickness measurement

[0259] 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.

[0260] Example 2: Effects of the active ingredients and combinations according to the invention on the KI67 factor

[0261] 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 carried out in triplicate.

[0262] The bioprinted skin samples were tested respectively with the following active ingredients and combinations (dissolved in the culture media):

[0263] - 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%).

[0264] An untreated negative control was carried out, without composition according to the invention.

[0265] Results

[0266] The results are shown in [Fig.l]. 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 makes it possible to 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.

[0267] Example 3: Effects of the active ingredients and combinations according to the invention on the thickness of the dermis

[0268] The effect of the active ingredients alone or in combinations according to the invention on the thickness of the dermis of the skin samples reconstructed by bioprinting was analyzed. Each test was carried out in triplicate.

[0269] The bioprinted skin samples were tested respectively with the following active ingredients and combinations (dissolved in the culture media):

[0270] - 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%).

[0271] An untreated negative control was carried out, without composition according to the invention.

[0272] Results

[0273] The results are shown in [Fig.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 for combating the thinning of the epidermis observed with age.

[0274] 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 in addition a bioactive fraction isolated from rose petals, makes it possible to obtain a synergistic anti-aging effect.

[0275] Example 4: Effects of the active ingredients alone or in combinations according to the invention on Collagen VII

[0276] The effect of the active ingredients alone or in combinations according to the invention, on the production of Collagen VII by the samples of skin reconstructed by bioprinting, was analyzed. Each test was carried out in triplicate.

[0277] The samples of skin reconstructed by bioprinting were tested respectively with the following active ingredients and combinations according to the invention (dissolved in the culture media):

[0278] - 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 0.21pg / 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%).

[0279] An untreated negative control was carried out, without active ingredients or combinations of active ingredients according to the invention.

[0280] Results

[0281] The results are shown in [Fig.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 VII collagen, essential for the good structure of the skin. Indeed, collagen VII is an essential component of the extracellular matrix, and gives the skin its firmness. This combination is therefore very advantageous to prevent the loss of expression of collagen VII observed with aging, and therefore combat the signs of aging.

[0282] Example 5: Cosmetic formulations according to the invention

[0283] 5.1 Composition in the form of an emulsion

[0284] Demineralized water qsp 100.0%

[0285] Glycols 20.0%

[0286] Preservatives 0.6%

[0287] Chelating agent 0.04%

[0288] Carbomer (Carbopol® 981) 0.3%

[0289] Sodium polyacrylate (Covacryl® MV60) 0.2%

[0290] Sodium Hydroxide 0.15%

[0291] Aqueous extract of rosewood * 1%

[0292] Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide

[0293] comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)

[0294] Palmitoyl tetrapeptide-7 (RIGIN® comprising

[0295] 0.05% peptide) 4% (i.e. 0.002% peptide)

[0296] Vegetable oil, esters 16%

[0297] Antioxidant 0.2%

[0298] Perfume concentrate 0.4%

[0299] Steareth-2 0.8%

[0300] Steareth-21 1.5%

[0301] * as described in Example 1

[0302] Applied to the skin, particularly the skin of the face, this composition provides tone, elasticity and firmness, the features are enhanced, the face is reshaped.

[0303] 5.2: Composition in the form of a rich face cream

[0304] Demineralized water qsp 100.0%

[0305] Glycols 13.0%

[0306] Carbomer (Carbopol® 981) 0.3%

[0307] Sodium polyacrylate (Covacryl® MV60) 0.2%

[0308] Sodium Hydroxide 0.15%

[0309] Aqueous extract of rosewood * 0.5% ZETA fraction ** 0.3%

[0310] Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide

[0311] comprising 0.05% peptide) 2% (i.e. 0.001% peptide)

[0312] Palmitoyl tetrapeptide-7 (RIGIN® comprising

[0313] 0.05% peptide) 3% (i.e. 0.0015% peptide)

[0314] Vegetable oil, esters 16%

[0315] Triglycerides of fatty acids 4%

[0316] Shea butter 1%

[0317] Antioxidant 0.2%

[0318] Perfume concentrate 0.4%

[0319] Steareth-2 0.8%

[0320] Steareth-21 1.5%

[0321] Preservatives qs

[0322] * and ** as described in Example 1

[0323] Applied to the skin of the face, this composition gives tone, elasticity and firmness, the features are enhanced, the face is reshaped.

[0324] 5.3: Composition in the form of a gel-serum for the eye contour

[0325] Purified water Qsp 100.00%

[0326] Glycols 13.0%

[0327] Preservatives 0.60%

[0328] Carbomer 0.80%

[0329] Glyceryl stearate citrate 0.70%

[0330] Lecithin and sodium acrylates copolymer (Lecigel PCR negative) 1.20%

[0331] Isostearyl isostearate 9.0%

[0332] Bis-diglyceryl polyacyladipate-2 (Softisan 649 MB) 1.0%

[0333] Silica 2.0%

[0334] Mother-of-pearl 1.0%

[0335] Centella asiatica extract 0.5%

[0336] Horse chestnut extract 0.1%

[0337] Aqueous extract of rosewood * 0.5%

[0338] Acetyl hexapeptide-8 (ARGIRELINE® Amplified peptide

[0339] comprising 0.05% peptide) 3% (i.e. 0.0015% peptide)

[0340] Palmitoyl tetrapeptide-7 (RIGIN® comprising

[0341] 0.05% peptide) 4% (i.e. 0.002% peptide) ZETA Fraction ** 0.3%

[0342] Tocopheryl acetate 0.10%

[0343] Rose fragrance 0.20%

[0344] * and ** as described in Example 1

[0345] After application to the eye contour of this gel-serum, it helps to smooth out wrinkles and fine lines, the eye area is firmed. The youthfulness of the eyes is restored with each application.

Claims

Claims

1. 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. A hexapeptide with INCI name: Acetyl Hexapeptide-8, and iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7.

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 ground rosewood 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 in addition at least one protease, c) incubation with stirring of the previously ground rosewood 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 proteins and fermentable sugars, and a solid phase, e) separation of the aqueous phase and optional addition of preservatives.

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. Cosmetic composition according to claim 1 or 2, characterized in

5.

6.

7. 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. 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. The tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with 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. Cosmetic composition according to claim 3, 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. 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, 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. 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. 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 skin care composition. Non-therapeutic cosmetic process for the care and / or makeup of the

8.

9. skin and / or lips comprising the application to said skin and / or said lips, in particular the skin, of a composition as defined in any one of claims 1 to 6. Cosmetic process according to claim 7, characterized in that the application of the composition to the skin and / or the lips makes it possible to prevent and / or reduce the signs of skin aging. Association of cosmetic active ingredients including: i. An aqueous extract of rosewood of the Evanrat variety, preferably summer rosewood, obtained by enzymatic extraction using a cosmetically acceptable polar solvent, ii. A hexapeptide with INCI name: Acetyl Hexapeptide-8, iii. A tetrapeptide Pal-GQPR-NH2 or Pal-GQPR-OH, in particular the tetrapeptide with INCI name: Palmitoyl Tetrapeptide-7, and iv. Optionally, in addition, 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. Non-therapeutic cosmetic use of the combination of cosmetic active ingredients as defined in claim 9, as an active combination for preventing and / or reducing 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.

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