Isolated bioactive fraction of Evanrat variety roses for soothing cosmetic use on the skin and / or lips

Abstract

The present invention relates to the cosmetic use of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as a soothing agent for the skin and / or lips.

Description

Title of the invention: Isolated bioactive fraction of roses of the Evanrat variety for soothing cosmetic use on the skin and / or lips FIELD OF INVENTION

[0001] The present invention relates to the use of an extract of Evanrat variety roses, also known as 'Jardin de Granville®' roses, in the form of an isolated bioactive fraction. The invention further relates to the use of a composition comprising said extract of Evanrat variety roses, or 'Jardin de Granville®' roses, in the form of an isolated bioactive fraction. According to the present invention, this extract is used for cosmetic purposes, as a soothing agent, and in particular to prevent and / or reduce irritation and micro-inflammation. STATE OF THE ART

[0002] The skin is the body's first protective barrier against the environment.

[0003] It contains a dense nervous system, essentially made up of free nerve endings penetrating to the outermost layers of the skin. These extensions of sensory neurons, or nerve fibers, play a major role in our perception of the environment and allow us to adapt to it more effectively.

[0004] Due to this direct contact with our environment, the skin is continually subjected to stresses of exogenous or endogenous origin that may induce non-pathological skin reactions. An exogenous stress can be of biological or chemical origin, for example xenobiotics, antigens, allergens, cosmetic products, compounds likely to cause skin irritation, peels; or of environmental origin (temperature, climate, UV radiation, air pollution, in particular heavy metals, ozone, cigarette smoke...) or even of mechanical origin (friction, shaving, cleaning...).

[0005] All these factors act on the skin and its nervous system. By disrupting its balance, they can generate phenomena of irritation and / or micro-inflammation, and ultimately contribute to the appearance of redness and / or itching; unsightly skin reactions that can also cause a feeling of discomfort.

[0006] To protect against these daily aggressions, there is a constant need to find new agents capable of preventing and / or reducing skin reactions due to exogenous and / or endogenous stresses and capable of soothing the skin.

[0007] In this context, the Applicant has demonstrated in vitro the soothing effects of an extract of roses of the Evanrat variety, in the form of a "bioactive serum fraction". Unexpectedly, the Applicant was able to show that this extract acts as a cutaneous neuroprotective agent.

[0008] In particular, this extract is capable of promoting the survival and / or neuritic growth of human sensory neurons located in the skin. This extract also increases the number of TRPV1 receptors involved in the transmission of pain and itching sensations. Furthermore, it reduces the release of CGRP neuropeptides and pro-inflammatory cytokines induced by irritant stress.

[0009] All of these effects make it possible to consider using this extract as a soothing agent for the skin and / or lips, in particular to prevent and / or reduce irritation and micro-inflammation. Description of the invention

[0010] A first object of the invention relates to the cosmetic use of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as a soothing agent for the skin and / or lips.

[0011] Preferably, said rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, is used as an agent intended to prevent and / or reduce the phenomena of irritation of the skin and / or lips.

[0012] Preferably, said irritation phenomena are induced in particular by at least one condition chosen from among atmospheric pollution, stress, irritant molecules, thermal shocks, and / or friction.

[0013] Preferably, said extract is present in a cosmetic composition in a content ranging from 0.001% to 50%, in particular from 0.01% to 20%, preferably from 0.01% to 10% and preferably again from 0.011% to 5% by weight of raw material in relation to the total weight of said cosmetic composition.

[0014] Preferably, said cosmetic composition further comprises at least one cosmetic adjuvant chosen from antioxidants, perfumes, vitamins, thickening agents, emollients, moisturizing agents, anti-aging agents, lifting agents, tightening agents, plumping agents, soothing agents, anti-pollution agents, brightening or depigmenting agents, fillers, mother-of-pearl and mixtures thereof.

[0015] Preferably, said cosmetic composition 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, especially for the skin, a balm, a gloss or a stick, especially for the lips.

[0016] Preferably, said extract or cosmetic composition is intended for application to subjects with irritated and / or stressed skin and / or lips.

[0017] Preferably, said extract or cosmetic composition is intended for topical application on the skin and / or lips and in particular on the skin of the face and / or neck.

[0018] Preferably, said extract is obtained by a process comprising the steps:

[0019] a) cleaning of plant material, maceration, pressing then mechanical separation of plant material to obtain an intracellular colloidal dispersion (ICD) and a material enriched in fibers (fraction A);

[0020] 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);

[0021] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0022] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "Bioactive Serum Fraction" and a D fraction (precipitate); and

[0023] e) Optionally, mixing the "bioactive serum fraction" with at least one preservative and / or stabilizer. DESCRIPTION OF THE FIGURES

[0024] [Fig. 1] extraction process implemented to obtain the extract used according to the invention DETAILED DESCRIPTION OF THE INVENTION Definitions

[0025] The term “bioactive fraction,” “bioactive serum fraction,” “serum fraction,” or “isolated bioactive fraction of roses” refers to an extract of roses of the Evanrat variety, or the 'Jardin de Granville®' rose, comprising the enzymes, proteins, sugars, ions, and other active molecules present in the cytosol of the cells composing the various plant tissues of the roses. The extract according to the invention is distinct from a cryoextract of rose petals as described in application FR3066388.

[0026] The following definitions are related to the steps of the process of preparing the extract according to the invention.

[0027] By "cleaning" is meant the removal of debris from fresh Granville® roses, and preferably fresh petals, before further processing, in a manner that avoids damaging the plant or removing valuable components. For example, it can be carried out by low-pressure rinsing with potable water under conditions where the Washing with runoff water would not contain any significant amount of plant pigments. Excess wash water is then removed from the washed plants.

[0028] “Maceration” refers to the process 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 maceration instruments include, but are not limited to, devices such as a crusher, a grinding wheel, or a grinder (e.g., a knife mill, a hammer mill, etc.). To avoid temperature-induced degradation of the plant material, the maceration step may include temperature monitoring and the selection of maceration parameters to ensure that there is no significant increase in the temperature of the plant material during this step.

[0029] “Pressing” means the separation of the liquid matter from fresh Granville® roses, and preferably from fresh petals, by the application of a mechanical force. This includes, but is not limited to, techniques such as ambient gravity drainage, pressing with a heavy object, the centrifugal force of a rotary expeller, the piston pressure of a hydraulic press, or rollers or a screw of a suitable type for a press.

[0030] By "fibre-enriched material" or "FEM" or "FEM" (Fibre Enriched Material), we mean a solid and / or semi-solid fraction enriched with fibers from fresh Granville® roses, preferably fresh petals, from which the liquid intracellular colloidal dispersion (ICD) has been removed by pressing.

[0031] By "intracellular colloidal dispersion" or "ICD" refers to 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 mainly of plant cell organelles, organelle fragments, and fiber-enriched residual material. The aqueous medium consists mainly of cytosols and vacuoles.

[0032] By "adjustment" is meant the modification of the activities of hydroxide and hydronium ions in the aqueous medium of the DCI or of aqueous fractions produced by subsequent treatment of the DCI, the hydronium ion activity remaining within the range found in viable plant cells (e.g., between pH 3 and pH 9). This alteration can be accomplished, for example, by a separation process such as S / L filtration, centrifugation, membrane filtration (with bipolar membranes, ultrafiltration and microfiltration, reverse osmosis), and / or by the addition of a weak acid or weak alkali / base, and by precipitation. The adjustment parameters are selected to be sufficient for a particular change in physicochemical parameters, such as pH or the surface potential at the electrolyte-air interface. Such adjustments facilitate subsequent destabilization and / or separation steps; or create the conditions for good preservation and stabilization.

[0033] By "destabilization" is meant the treatment of the adjusted DCI using electromagnetic waves to transiently modify physical properties (such as e'0, which is the real component of the low-frequency dielectric constant). It was unexpectedly found that particular modifications degrade the stability of the DCI by causing the agglomeration and / or aggregation of particles into assemblies that are sufficiently large and stable to permit and / or improve, after separation into fractions, certain desirable properties.

[0034] “Separation” means the separation of solid and / or semi-solid particles and non-aqueous liquid droplets from an aqueous liquid by exploiting particle density and / or size. This includes, but is not limited to, techniques such as draining, filtration (including pressure gradient filtration), skimming, ambient gravity sedimentation, decantation, centrifugation, or a combination thereof. Preferably, continuous flow mechanical separation is used, but this does not preclude batch processing. “Separation 1” and “Separation 2” refer to the respective process steps, carried out with respective parameters.

[0035] The term "supernatant" refers to an aqueous material from which particles have been separated. "Supernatant A" and "Supernatant B" denote the supernatants resulting from the respective separation steps of the process.

[0036] The term "precipitate" refers to the particles from which an aqueous material has been separated. "Fraction B" and "Fraction C" denote the precipitates resulting from the respective separation steps of the process.

[0037] The terms "serum fraction of fresh Granville® roses" and "serum fraction of fresh Granville® rose petals" refer to compositions produced by the process as shown above and in [Fig. 1] without preservatives and / or stabilizers added to protect the ingredient composition against environmental factors such as temperature, atmosphere (e.g., oxygen), light and microorganisms.

[0038] By "preservatives and / or stabilizers" is meant substances which, when added to a "serum fraction of fresh Granville® roses", preferably a "serum fraction of fresh Granville® rose petals", protect it against environmental factors such as temperature, atmosphere (e.g. oxygen), light and microorganisms. Suitable substances include ingredients may include, without limitation, a preservative, a stabilizer and / or a mixture thereof.

[0039] The term “Granville® rose serum” or “extract of fresh Granville® roses” as used herein means a combination of a fraction of serum from fresh Granville® roses and preservatives and / or stabilizers.

[0040] 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.

[0041] Rose extract: Bioactive fraction isolated from roses and preparation process

[0042] The present invention relates to the cosmetic use of a rose extract obtained by a specific process, notably involving a destabilization step using electromagnetic waves. The extract obtained from this process has superior activity compared to previously known aqueous rose extracts.

[0043] This extract is also called “Granville Rose Serum®” Plant material

[0044] In the context of the present invention, plant material will be used from roses of the Evanrat variety, also called 'Jardin de Granville®' roses, and preferably even more so from rose petals of the Evanrat variety, or 'Jardin de Granville®' roses.

[0045] The 'Jardin de Granville®' rose is a hybrid variety offered exclusively by "Roses anciennes André Eve SAS" and protected by Plant Variety Certificate No. 20110345, with the species name Rosa L. and the variety name EVANRAT. This bush rose belongs to the group of modern hybrids, which, from May to October, are continuously covered in roses, thus demonstrating excellent repeat flowering.

[0046] The extract used in the present invention is therefore an extract of roses, more particularly an extract of roses of the Evanrat variety, or 'Jardin de Granville®' roses.

[0047] The invention preferably uses selected roses, whose properties are preserved by an organic environment and farming method.

[0048] The rose extract used in the present invention can be prepared from fresh, frozen, freeze-dried roses, or any mixture thereof. In the context of the invention, fresh roses are preferably used.

[0049] Fresh, i.e., living plants have maximum metabolic activity, thus representing an optimal source for capturing the full spectrum of natural complexes and compounds and preserving their properties. Therefore, the use of fresh roses is preferred. Plant viability can be verified, in particular, by measuring chlorophyll fluorescence.

[0050] Thus, the rose extract used in the present invention is a rose extract from the variety Evanrat, or roses 'Jardin de Granville®', in the form of an isolated bioactive fraction, obtained from fresh roses.

[0051] Two types of roses are distinguished according to the time of year they are harvested: - Winter roses, generally harvested between November and April, - Summer roses, generally harvested between May and October.

[0052] According to one particular embodiment, the invention uses summer roses, in particular summer rose petals. According to another embodiment, winter roses, in particular winter rose petals, will preferably be used.

[0053] The extract used according to the invention can be prepared from different parts of the plant, it can therefore be a leaf extract, a bud extract, a flower (petal) extract, a sepal extract, a wood (stem) extract, a root extract or mixtures thereof.

[0054] Advantageously, the extract used according to the invention is a petal extract.

[0055] According to a preferred embodiment, the extract used is an extract of fresh petals, and more particularly an extract of fresh petals of roses of the Evanrat variety, or 'Jardin de Granville®' roses.

[0056] 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.

[0057] Rich in monosaccharide sugars means 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 selected from fructose, glucose and sucrose.

[0058] By mineral-rich, 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 selected from potassium, calcium and sodium.

[0059] Advantageously, calcium improves epidermal differentiation and strengthens the skin's structure, while potassium enhances skin hydration and boosts energy assimilation. Phyto-sugars (fructose, glucose, sucrose) help to saturate skin cells with energy.

[0060] The extract of Evanrat variety roses, or 'Jardin de Granville®' roses, preferably the extract of Evanrat variety rose petals or 'Jardin de Granville®' roses, used in the context of the present invention is in the form of an isolated bioactive fraction and more particularly in the form of a bioactive serum fraction. Compared to known aqueous extracts of the prior art (e.g., Aqueous cryo-extract of rose flowers (petals) (application FR3066388) is a higher concentration of natural compounds, including sugars such as fructose and glucose, as well as polyphenols and minerals. Therefore, due to the specific process used to obtain this extract, the rose extract used in the context of the invention is distinct from rose water. Extraction process

[0061] The extract of fresh roses of the Evanrat variety or 'Jardin de Granville®' roses used in the context of the present invention, also called "Granville Rose Serum", is advantageously obtained by implementing the extraction process described below as well as in [Fig. 1].

[0062] Preferably, fresh roses of the Evanrat variety or 'Jardin de Granville®' roses are used as plant material. According to a preferred method, this refers to fresh petals of roses of the Evanrat variety or 'Jardin de Granville®' roses.

[0063] Advantageously, said process does not require the addition of any solvent or exogenous liquid.

[0064] The process implemented to obtain the extract used according to the invention comprises the main steps of:

[0065] a) cleaning of plant material, maceration, pressing then mechanical separation of plant material to obtain an intracellular colloidal dispersion (ICD) and a material enriched in fibers (fraction A);

[0066] b) “Treatment A” then mechanical separation of the Intracellular Colloidal Dispersion (ICD) to obtain the Supernatant A and a Membrane Fraction (Fraction B);

[0067] c) “Treatment B” then mechanical separation of the Supernatant A to obtain the Supernatant B and the Fraction C (Cytoplasmic Fraction);

[0068] d) “Treatment C” then mechanical separation of the Supernatant B to give a “Bioactive serum fraction” and a D fraction (Precipitate); and

[0069] e) Optionally, mixing the "bioactive serum fraction" with at least one preservative and / or stabilizer.

[0070] The extract of fresh roses, preferably fresh rose petals, of the Evanrat variety or 'Jardin de Granville®' rose obtained by this process, is a "bioactive serum fraction" or "bioactive fraction". This is the extract used in the context of the present invention.

[0071] 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, CN1929851B, U.S. patents No. 8,734,861 and No. 7,473,435; U.S. patent application No. 16 / 078925.

[0072] Advantageously, this extraction process does not involve solvents or- Organic compounds, such as hexane, can cause numerous safety problems for facilities and personnel, as well as issues with human health and environmental protection. This technology reduces emissions of volatile organic compounds (VOCs). It is a clean process that extracts high-quality proteins, sugars, and other co-products directly into an aqueous extract without the addition of solvents, making them directly usable in cosmetics.

[0073] Preferably, the flowers of roses (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 in order to prevent disruption of the cellular structure of the flowers. The viability of the collected plants can be tested using an OS5p multimode chlorophyll fluorometer (Opti-Sciences Inc, Hudson, NH, USA).

[0074] According to a particular embodiment, 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 immediately placed 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.

[0075] 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°C and 20°C and preferably at a temperature between 2°C and 6°C until harvesting is complete. Step a

[0076] 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 at 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 pressing to extract the contents of the liquid intercellular colloidal dispersion (ICD) from the fiber-enriched material (“Fraction A”).

[0077] 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).

[0078] The INN typically comprises 2% to 20% dry matter, preferably 4% to 16% dry matter, and even more preferably 6% to 12% dry matter.

[0079] At this stage, the INN can be frozen for storage. Typically, it is frozen at - 20°C. Step b

[0080] When the INN has been frozen for storage, it must first be gently thawed. Typically, it is placed to thaw at 4°C or in ice.

[0081] “Treatment A” is carried out by destabilizing the DCI with electromagnetic waves produced by magnetrons operating at a frequency between 2.45 and 5.8 GHz. The parameters of the destabilization treatment are set to reduce 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, organelle fragments, residual fibrous material) into assemblies large and stable enough to allow and / or improve mechanical separation.

[0082] Indeed, the resulting intracellular colloidal suspension is considered 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 each other unless they have sufficient 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 the potential energy of the particles as a function of their parameters, their distance from each other, and the characteristics of the continuous phase.Modifying the values ​​of variables affecting the repulsive force impacts the stability of the dispersion. Under normal colloidal stability conditions, 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 modified conditions during treatment A allow the repulsive force of the double layer to decrease to the point where the potential energy barrier disappears, and the particles can approach and agglomerate freely.

[0083] 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., softw journal 2017).

[0084] Preferably, following "treatment A", the mechanical separation step of the ICD is carried out by centrifugation in order to produce the "Supernatant A" and the " Fraction B.

[0085] Typically, the "supernatant A" has a turbidity of less than about 100 NTU.

[0086] The “fraction B” typically comprises 10% to 30% dry matter, preferably 13% to 27% dry matter, and even more preferably about 15.0% to 25.0% dry matter. Step c

[0087] The "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.

[0088] Following the "treatment B", the mechanical separation step of the "Supernatant A" is preferably carried out by centrifugation in order to produce the "Supernatant B" and the "Fraction C".

[0089] The “fraction C” typically comprises from 5% to 25% dry matter, preferably from 8% to 22% dry matter, and even more preferably about 10.0% to 20.0% dry matter. Step d

[0090] The "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 below 4.5 is obtained. Typically, a citric acid solution will be used as the preferred acid.

[0091] Following the "treatment C", the mechanical separation step of the "Supernatant B" is preferably carried out by centrifugation in order to produce the "serum fraction of fresh Granville® roses", preferably the "serum fraction of fresh Granville® rose petals" (Non-Preserved Extract) and the "Fraction D".

[0092] The “serum fraction of fresh Granville® rose petals” typically comprises 2% to 20% dry matter, preferably 4% to 16% dry matter, and even more preferably 6.0% to 10.0% dry matter. Step e

[0093] According to some 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 the fresh Granville® rose petal extract or "Granville® Rose Petal Serum".

[0094] Particularly suitable stabilizing agents may include, without limitation, a preservative, a stabilizer, and / or mixtures thereof. Preservatives and stabilizers suitable for use in the present invention include These 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 comprise at least one preservative, at least one stabilizer, at least one antioxidant, or mixtures thereof.

[0095] Thus, according to a preferred embodiment, the process implemented to obtain the extract used according to the invention comprises the steps of:

[0096] a) cleaning of plant material, maceration, pressing then mechanical separation of plant material to obtain an intracellular colloidal dispersion (ICD) and a material enriched in fibers (fraction A);

[0097] 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);

[0098] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0099] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "Bioactive Serum Fraction" and a D fraction (precipitate); and

[0100] e) Optionally, mixing of the “bioactive serum fraction” with at least one preservative and / or stabilizer.

[0101] Preferably, the process used to obtain the extract used according to the invention comprises the steps of:

[0102] a) cleaning of fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, maceration, pressing then mechanical separation of the roses to obtain an intracellular colloidal dispersion (INN) and a material enriched in fibers (fraction A);

[0103] 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);

[0104] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0105] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "serum fraction of fresh Granville® roses" and a fraction D (precipitate); and

[0106] e) Optionally, mixing of the “serum fraction of fresh Granville® roses” with at least one preservative and / or stabilizer.

[0107] Preferably, the process implemented to obtain the extract used according to The invention comprises the steps of:

[0108] a) cleaning of fresh rose petals of the Evanrat variety, or 'Jardin de Granville®' roses, maceration, pressing then mechanical separation of the roses to obtain an intracellular colloidal dispersion (INN) and a material enriched in fibers (fraction A);

[0109] 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);

[0110] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0111] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "serum fraction of fresh Granville® rose petals" and a fraction D (precipitate); and

[0112] e) Optionally, mixing of the “fragment of fresh Granville® rose petal serum” with at least one preservative and / or stabilizer.

[0113] Preferably, the invention relates to the use of an extract in the form of a bioactive fraction isolated from fresh roses of the Evanrat variety or 'Jardin de Granville®' roses, characterized in that it is obtained by a process comprising the steps:

[0114] a) cleaning of plant material, maceration, pressing then mechanical separation of plant material to obtain an intracellular colloidal dispersion (ICD) and a fiber-enriched material (fraction A);

[0115] 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);

[0116] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0117] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "Bioactive Serum Fraction" and a D fraction (precipitate); and

[0118] e) Optionally, mixing of the “bioactive serum fraction” with at least one preservative and / or stabilizer.

[0119] Preferably, the invention relates to the use of an extract in the form of a bioactive fraction isolated from fresh roses of the Evanrat variety or 'Jardin de Granville®' roses, characterized in that it is obtained by a process comprising the steps:

[0120] a) cleaning fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, maceration, pressing then mechanical separation of the roses to obtain a dispersion intracellular colloidal (ICD) and a fiber-enriched material (fraction A);

[0121] 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);

[0122] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0123] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "serum fraction of fresh Granville® roses" and a fraction D (precipitate); and

[0124] e) Optionally, mixing of the “serum fraction of fresh Granville® roses” with at least one preservative and / or stabilizer.

[0125] Preferably, the invention relates to the use of an extract in the form of a bioactive fraction isolated from fresh roses of the Evanrat variety or 'Jardin de Granville®' roses, characterized in that it is obtained by a process comprising the steps:

[0126] a) cleaning of fresh rose petals of the Evanrat variety, or 'Jardin de Granville®' roses, maceration, pressing then mechanical separation of the roses to obtain an intracellular colloidal dispersion (INN) and a material enriched in fibers (fraction A);

[0127] 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);

[0128] c) adjusting the pH level in the supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, then mechanically separating the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0129] d) adjusting the pH level in supernatant B until a pH below 4.5 is obtained, then mechanically separating supernatant B to give a "Granville® fresh rose petal serum fraction" and a D fraction (precipitate); and

[0130] e) Optionally, mixing of the “fragment of fresh Granville® rose petal serum” with at least one preservative and / or stabilizer.

[0131] According to a particular method, the rose extract, preferably rose petal extract, used according to the invention comprises water in a content ranging from 90 to 94%, and a dry extract content ranging from 6 to 10%. This rose extract, preferably rose petal extract, in the form of an isolated bioactive fraction, used according to the invention, is designated by its INCI name: “Rosa Hybrid Flower Extract”.

[0132] According to another particular embodiment, the rose extract, preferably rose petal extract, used 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.

[0133] The extract of roses of the Evanrat variety, or 'Jardin de Granville®' roses, in particular of rose petals of the Evanrat variety, or 'Jardin de Granville®' roses, used according to the invention, comprises in particular amino acids (aspartic acid, tyrosine, arginine) total sugars (glucose, fructose), minerals and other co-products, which when applied to a keratinous material, in particular the skin, provides a benefit or an improvement in the appearance of the keratinous material, such as promoting and / or improving skin nutrition, promoting and / or improving the skin barrier function, promoting and / or improving the rhythmic process of skin cells, preventing and / or slowing down skin aging.

[0134] Furthermore, the Applicant has been able to show that the extract used according to the invention acts as a cutaneous neuroprotective agent. In particular, it soothes the skin and / or lips, notably by preventing and / or reducing irritation and micro-inflammation. Composition and dosage

[0135] The present invention also relates to the use of a composition comprising at least one extract in the form of an isolated bioactive fraction of fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses according to the invention, as defined above.

[0136] Preferably, said composition comprises at least one extract in the form of an isolated bioactive fraction of fresh rose petals of the Evanrat variety, or 'Jardin de Granville®' roses, as defined above.

[0137] The composition used in the present invention is preferably a cosmetic composition.

[0138] By "cosmetic composition" we mean any composition intended for cosmetic, i.e. aesthetic purposes, which may be brought into contact with the superficial parts of the human body and more particularly with keratinous materials, in particular the skin and / or lips, in particular the skin of the face and / or neck.

[0139] By “keratinous material” we mean the skin and / or its appendages, and the lips, in particular the skin of the face and / or body and the lips.

[0140] The present invention therefore relates to the use of a cosmetic composition comprising, in a physiologically acceptable medium, an effective quantity of at least one extract in the form of a bioactive fraction isolated from fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses as defined above.

[0141] By "physiologically acceptable medium" is meant any excipient suitable for topical use, in contact with keratinous materials, without risk of toxicity, incompatibility, instability and / or allergic response.

[0142] By "effective quantity" is meant the minimum quantity of "Jardin de Granville®" rose extract according to the invention which is necessary to obtain the beneficial effect according to the invention, namely a beneficial effect consisting of soothing the skin and / or lips, in particular preventing and / or reducing the phenomena of irritation, and / or preventing and / or reducing the phenomena of micro-inflammation.

[0143] The cosmetic composition according to the invention is a care composition for keratinous materials, in particular of the skin and / or lips and especially of the skin of the face and / or neck.

[0144] According to an advantageous embodiment, the extract in the form of an isolated bioactive fraction of fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, is present in the cosmetic composition, used according to the invention, in a content ranging from 0.001 to 50%, in particular from 0.01 to 20%, preferably from 0.01 to 10% and even more preferably from 0.011 to 5% by weight of raw material in relation to the total weight of said composition.

[0145] The physiologically acceptable medium generally represents from 1 to 99% by weight, relative to the total weight of said composition.

[0146] The cosmetic composition used according to the invention generally comprises, in addition to the "Jardin de Granville®" rose extract and the physiologically acceptable medium, one or more acceptable cosmetic excipients from among those known to the person skilled in the art in order to obtain a composition for topical application, 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 powder.

[0147] According to a particular mode, the cosmetic composition used according to 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, especially for the skin.

[0148] In a preferred embodiment, said composition used according to the invention is in the form of a cream or a serum, particularly for the skin.

[0149] In another preferred embodiment, said composition used according to the invention is in the form of a balm, a gloss or a stick, in particular for the lips.

[0150] The cosmetic composition used according to the invention may be in any pharmaceutical form suitable for topical application to the skin and / or lips, and in particular to the skin of the face and / or neck, comprising the "Jardin de Granville®" rose extract, preferably the "Jardin de Granville®" rose petal extract, and at least one cosmetic adjuvant chosen from among antioxidants, perfumes, vitamins, thickening agents, emollients, moisturizing agents, anti-aging agents, lifting agents, firming agents, plumping agents, soothing agents, anti-pollution agents, brightening or depigmenting agents, charges, mother-of-pearl and their mixtures.

[0151] Also, according to a particular embodiment of the invention, the cosmetic composition according to the invention may further comprise at least one cosmetic adjuvant chosen from the group consisting of: antioxidant agents, emollient agents, moisturizing agents, anti-aging agents, perfumes, and mixtures thereof.

[0152] Depending on the nature of the composition, one or more cosmetically acceptable excipients will be selected from emulsifiers, polymers, surfactants, rheology agents, electrolytes, pH adjusters, antioxidants, preservatives, colorants, and mixtures thereof.

[0153] By way of particular example, the cosmetic composition according to the invention may include gelling agents, antioxidants, preservatives and mixtures thereof.

[0154] The cosmetic composition according to the invention may further comprise a fatty (solid fats) or oily phase.

[0155] The term "oil phase" refers to an oil or a mixture of oils, whether miscible or immiscible. For the purposes of this invention, "oil" refers to 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.

[0156] An oily phase according to the invention may include natural, hydrocarbon, silicone oils, and mixtures thereof.

[0157] The oily or fatty phase content in the cosmetic composition of the invention will generally range from 0.2% to 45%, preferably from 0.5% to 30%, and even more preferably from 2% to 25% by weight relative to the total weight of said composition. Cosmetic and other uses

[0158] In the context of the invention, the extract described above and the cosmetic compositions containing it are not intended for therapeutic use in sick subjects. In particular, they are not intended for use in the prevention or treatment of phenomena and / or symptoms that are pathological in nature and would require medical treatment in sick subjects.

[0159] The present invention relates to the cosmetic use of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as a soothing agent for the skin and / or lips.

[0160] The term "soothing agent" refers to an agent capable of preventing and / or reducing skin reactions induced by exogenous and / or endogenous stress, in particular by protecting the skin against the effects of irritant stress. Irritant stressors within the meaning of the invention include, in particular, UV radiation, air pollution, stress, allergenic or irritating molecules, thermal shock, and / or friction, and preferably air pollution, stress, and ir- ritantes, thermal shocks, and / or friction.

[0161] The Applicant has indeed shown that the rose extract in the form of an isolated bioactive fraction obtained from fresh roses of the Evanrat variety and used according to the invention had a cutaneous neuroprotective effect.

[0162] The term “cutaneous neuroprotective agent” or “cutaneous neuroprotective agent” refers to an agent capable of preventing and / or reducing skin reactions induced by exogenous and / or endogenous stress, in particular by protecting human cutaneous sensory neurons against the effects of irritant stress. Irritant stressors within the meaning of the invention include, in particular, UV radiation, air pollution, stress, allergenic or irritating molecules, thermal shock, and / or friction, and preferably air pollution, stress, irritating molecules, thermal shock, and / or friction.

[0163] In particular, the term "cutaneous neuroprotective agent" means that the extract used promotes the survival and / or neuritic growth of human sensory neurons in the skin. It may also be referred to as a "cutaneous neuro-soothing agent".

[0164] According to a particular aspect, the invention also relates to the cosmetic use of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as an agent intended to prevent and / or reduce the phenomena of irritation of the skin and / or lips.

[0165] According to the present invention, "irritation" means that keratinous materials exhibit increased sensitivity to external stimuli such as wind or extreme temperatures, resulting in a feeling of discomfort. Irritated keratinous materials may have a reddish appearance, may also be slightly cracked, and may give an itchy sensation.

[0166] According to the present invention, the term "irritation phenomena" refers to phenomena of moderate intensity and whose scope is limited to the field of cosmetology. In particular, the irritation phenomena referred to are not pathological in nature and do not require medical treatment.

[0167] In particular, the terms "agent intended to prevent and / or reduce irritation" or "cutaneous neuroprotective agent intended to prevent and / or reduce irritation" mean that the extract used reduces the number of TRPV1 receptors responsible for transmitting pain and itching sensations. It also reduces the release of CGRP neuropeptides induced by irritant stress.

[0168] Another particular aspect of the invention relates to the cosmetic use of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as an agent intended to prevent and / or reduce the phenomena of micro-inflammation of the skin and / or lips.

[0169] According to the invention, "micro-inflammation" or "chronic micro-inflammation of the skin" refers to invisible (silent) inflammatory processes occurring locally in the skin. Over time, these repeated micro-inflammatory events contribute to skin aging.

[0170] According to the present invention, the term "micro-inflammation phenomena" refers to phenomena of moderate intensity and limited to the field of cosmetology. In particular, the micro-inflammation phenomena referred to are not pathological in nature and do not require medical treatment.

[0171] In particular, by "cutaneous neuroprotective agent intended to prevent and / or reduce micro-inflammation phenomena" it is understood that the extract used reduces the release of pro-inflammatory cytokines induced by irritant stress on the skin and / or lips.

[0172] Thus, the "keratinous materials" according to the invention are healthy keratinous materials (subjects "healthy"), that is to say, not exhibiting any disorders or abnormalities that would indicate a pathological condition (subjects "unhealthy", suffering from a pathology). The terms "healthy skin" and / or "lips" or "skin" and / or "lips" will be used interchangeably throughout the rest of the description.

[0173] In the context of the invention, said irritation phenomena and / or said micro-inflammation phenomena may be induced in particular by at least one condition selected from among UV radiation, air pollution, stress, allergenic or irritating molecules, thermal shock, and friction. They are preferably induced by at least one condition selected from among air pollution, stress, irritating molecules, thermal shock, and friction.

[0174] Thus, in the context of the invention, said extract is intended for application to subjects with irritated and / or stressed skin and / or lips.

[0175] In particular, said extract is intended for topical application on the skin and / or lips and in particular on the skin of the face and / or neck.

[0176] The extracts of fresh roses of the Evanrat variety, or 'Jardin de Granville®' roses, in the form of an isolated bioactive fraction, used for carrying out these processes and uses according to the invention are as described above in this application. In a particular and preferred embodiment, the rose extracts in the form of an isolated bioactive fraction are extracts in the form of an isolated bioactive fraction of rose petals, in particular roses of the Evanrat variety, and in particular roses from Jardin de Granville® rose bushes.

[0177] Other specific aspects of the invention relate to:

[0178] A rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, for its use as a cutaneous neuroprotective agent intended to soothe the skin and / or lips.

[0179] An extract of roses in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, for its use as a cutaneous neuroprotective agent intended to prevent and / or reduce the phenomena of irritation of the skin and / or lips.

[0180] An extract of roses in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, for its use in preventing and / or reducing skin and / or lip irritation.

[0181] An extract of roses in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, for its use as a cutaneous neuroprotective agent intended to prevent and / or reduce micro-inflammation phenomena of the skin and / or lips.

[0182] An extract of roses in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, for its use in preventing and / or reducing micro-inflammation of the skin and / or lips.

[0183] A method for soothing the skin and / or lips comprising administering an effective amount of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as described above, or a cosmetic composition comprising it.

[0184] A method for preventing and / or reducing skin and / or lip irritation phenomena comprising administering an effective amount of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as described above, or a cosmetic composition comprising it.

[0185] A method for preventing and / or reducing micro-inflammation of the skin and / or lips comprising administering an effective amount of a rose extract in the form of an isolated bioactive fraction, obtained from fresh roses of the Evanrat variety, as described above, or a cosmetic composition comprising it.

[0186] The invention will now be illustrated in the following non-limiting examples. Unless otherwise indicated, percentages are expressed by weight relative to the total weight of the composition. EXAMPLES Assets valued

[0187] Zeta-fraction of Granville rose® according to the invention

[0188] Fresh rose petals of the Evanrat variety are used as plant material or "Jardin de Granville®" rose.

[0189] The Granville® rose Zeta-fraction according to the invention is a bioactive fraction of rose petals, obtained in particular according to the following protocol:

[0190] a) cleaning 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,

[0191] a2) maceration, pressing and 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) of the fiber-enriched material ("Fraction A");

[0192] bl) destabilization of the DCI with electromagnetic waves produced from magnetrons operating at a frequency between 2.45 and 5.8 GHz,

[0193] b2) centrifugation of the intracellular colloidal dispersion (ICD) to obtain the supernatant A and a Membrane Fraction (fraction B);

[0194] cl) adjustment of the pH level in the supernatant A by an alkali (i.e., potassium carbonate), in order to obtain a pH ranging from 6 to 7,

[0195] c2) mechanical separation of the supernatant A to obtain the supernatant B and the fraction C (cytoplasmic fraction);

[0196] dl) adjusting the pH level in the supernatant B with citric acid to obtain a pH below 4.5,

[0197] d2) mechanical separation of the supernatant B to give a “serum fraction of fresh petals of Granville® roses and a D fraction (precipitate); and

[0198] e) mixture of the “fragment of fresh rose petal serum from Granville®” with potassium sorbate and sodium benzoate.

[0199] Granville® Rose Petal Serum (also referred to as "Zeta-fraction" or "Zf-fraction" in the following examples) contains 8% dry matter (active ingredient), 91.5% water by weight, 0.15% potassium sorbate by weight, and 0.3% sodium benzoate by weight. These amounts are expressed as a percentage of the total weight of the extract. The INCI name for this extract is Rosa Hybrid Flower Extract, Potassium Sorbate, and Sodium Benzoate. Positive test

[0200] Oxytocin (Sigma) was used as a positive control.

[0201] Oxytocin is a neuropeptide hormone known to reduce stress and regulate strong emotions. Its role in skin cells has been demonstrated by gene inactivation: oxytocin has a protective role, primarily as an antioxidant and anti-inflammatory agent. It prevents cellular senescence by reducing the release of pro-inflammatory mediators by senescent cells (dermal fibroblasts from donors of different ages) which are harmful to neighboring cells. via the activation of a defense signaling channel linked to its OXTR receiver.

[0202] The effect of the active ingredients was tested at the following concentrations:

[0203] - Zeta-fraction serum: concentrations 0.1%, 0.03% and 0.01%.

[0204] - Oxytocin: concentrations 0.2 mg / mL, 0.06 mg / mL, 0.02 mg / mL.

[0205] Example 1: Protective effect of the Zeta-fraction of Granville® rose on the cutaneous nervous system

[0206] The cutaneous neuroprotective effect of the active ingredients was evaluated, in particular by studying the release of CGRP (calcitonin gene-related peptide) neuropeptides, the release of pro-inflammatory cytokines, and the physiology of human sensory neurons in co-culture with innervated epidermis. These studies were conducted in the absence or presence of capsaicin stress. Cell cultures

[0207] The cell model used in the examples below was set up as follows:

[0208] Sensory neurons are derived from human induced pluripotent stem cells (hiPS cells), themselves obtained from human fibroblasts. The hiPS cells were seeded in sensory neuron differentiation medium (Ml) in 6-well plates pre-coated with a thin layer of Matrigel® (Dutscher). The cells were cultured for 6 days at 37°C and 5% CO2. The culture medium was changed every 2 days.

[0209] Under these conditions, hiPS differentiate into human sensory neurons. They express, in particular, the receptors for capsaicin (TRPV1), NGF (TrkA), and opiates (MOPr). They also synthesize Substance P and CGRP in their cytoplasm.

[0210] At this stage, the cells were dissociated and then seeded into 96-well plates covered with a thin layer of Matrigel® at a rate of 20,000 cells per well in differentiation medium.

[0211] After 9 days of culture, the sensory neurons were dissociated using Accutase and seeded into 24-well plates coated with a thin layer of Matrigel® at a rate of 120,000 cells per well, in differentiation medium. After 24 hours, reconstructed epidermis (StratiCell) (10-day maturation stage) was deposited over the neurons.

[0212] After 3 days of co-culture, the active ingredients were grown at 3 different concentrations. [3-Endorphin (Bachem)] was used as a positive control. Indeed, [3-Endorphin] is a peptide naturally produced by the body that acts as a neurotransmitter on opioid receptors. [3-Endorphin] has analgesic properties, notably by making neurons more sensitive to opioids.

[0213] The incubation conditions were as follows:

[0214] - Control environment;

[0215] - Control medium + Oxytocin at 0.2mg / ml, 0.06mg / ml or 0.02mg / ml;

[0216] - Control medium + Zeta-fraction serum at 0.1%, 0.03% or 0.01%;

[0217] - Control medium + [3-endorphin at 1 pM.

[0218] This treatment was repeated after 5 days and 8 days of culture.

[0219] After a total incubation period of 8 days, the cells were either stimulated or not stimulated for 30 minutes with capsaicin (Sigma) at a concentration of 1 OpM. Capsaicin, also known as 8-methyl-N-vanillyl-6-nonenamide, belongs to the vanilloid family. This irritant molecule is an active component of chili peppers (Capsicum). Capsaicin binds to TRPV1 (Transient Receptor Potential Vanilloid 7) receptors on sensory neurons, where it generates a sensation of discomfort. TRPV1 receptors are activated by thermal, mechanical, or chemical stimulation. Their response threshold is lower in individuals with sensitive skin.

[0220] Each culture condition was evaluated in triplicate.

[0221] At the end of the capsaicin stimulation, the supernatants were recovered and stored at -80°C for CGRP assay.

[0222] The co-cultures (neurons and epidermis) were then re-incubated in the presence of the active ingredients (oxytocin or Zeta-fraction) or [3-endorphin] for 6 hours and then for an additional 24 hours. After 6 hours and 24 hours, 50 pL of supernatant were collected for the measurement of cytokines II-1 [3, IL-6 and IL-8. GCRP assay

[0223] CGRP was measured using the anti-CGRP ELISA kit (Antibody-online), and the amounts of CGRP released in the presence of the active ingredients were compared to the control medium. Cytokine assay

[0224] Cytokines II-1 [3, IL-6 and IL-8 (BD Bioscience) were measured by flow cytometry and the amounts released in the presence of the active ingredients were compared to the control medium. Immunodetection

[0225] Neurons were labeled with an anti-3-tubulin antibody (Abcam), specifically to label neuronal processes, an anti-TRPV1 antibody (Sigma), and an anti-oxytocin receptor antibody (Sigma) for 12 hours at 4°C. These antibodies were detected by secondary antibodies coupled to different fluorochromes (Fisher Scientific) for 1 hour at room temperature and protected from light. Nuclei were identified using the Hoechst fluorescent marker (Sigma).

[0226] Per culture well, 2 sets of 20 photographs were taken with the automated microscope (In Cell 2200; GE Healthcare) at 20x magnification. A count of sensory neurons, a measurement of the length of neuronal processes, of TRPV1 receptor and oxytocin receptor expression were performed and the results were compared to the control condition.

[0227] At the end of the culture period (i.e., after 9 days of treatment), the epidermis was morphologically analyzed. The structure and thickness of each epidermis were compared to those of the control condition (treated with control medium). Statistics

[0228] In all the results presented below, statistical tests were performed using a One Way ANOVA test. Significance values ​​are represented as follows: * p<0.05; **p<0.01; *** p<0.001; **** p<0.0001 Results

[0229] Effect of the Zeta-fraction of Granville rose® on the release of neurovegetids CGRP

[0230] The neuropeptide CGRP (calcitonin gene-related peptide) is the first relay in the transmission of nociceptive information. A pro-inflammatory neurotransmitter, it is a mediator of pain.

[0231] CGPRs were measured after 8 days of treatment with the active ingredients + 30 minutes of treatment with capsaicin. The values ​​are normalized with respect to the number of sensory neurons.

[0232] The results of the assay are summarized in Tables 1 to 3.

[0233] [Tables 1] Treatment Variation / Untreated Control Beta-endorphin 1 pM -4% Zeta-fraction 0.1% -8% Zeta-fraction 0.03% -4% Zeta-fraction 0.01% +9% Oxytocin 0.2 mg / mL -16% Oxytocin 0.06 mg / mL +2% Oxytocin 0.02 mg / mL +7%

[0234] At baseline (in the absence of stress) no significant effect on CGRP neuropeptide release is observed.

[0235] [Tables2] Treatment Variation / Untreated Control Control Capsaicin lOpM +56%*

[0236] [Tables3] Treatment: active ingredients + CAPSAICIN 10 pM Variation / Capsaicin Control Capsaicin + Beta-endorphin 1 pM -49%** Capsaicin + Zeta-fraction 0.1% -74%**** Capsaicin + Zeta-fraction 0.03% -61%*** Capsaicin + Zeta-fraction 0.01% -69%*** Capsaicin + Oxytocin 0.2 mg / mL -39%* Capsaicin + Oxytocin 0.06 mg / mL -8% Capsaicin + Oxytocin 0.02 mg / mL -9%

[0237] It is observed that treatment with capsaicin results in a significant increase (+56%*) in the release of neuropeptide CGRP relative to the number of sensory neurons.

[0238] Treatment of the co-culture for 8 days with beta-endorphin at IpM before 30 minutes of capsaicin stress results in a significant decrease in the release of neuropeptide CGRP reported per number of neurons compared to the capsaicin control.

[0239] Treatment of the co-culture for 8 days with the active Zeta-fraction at concentrations of 0.1%, 0.03%, or 0.01% prior to capsaicin stress resulted in a significant decrease in the release of neuropeptide CGRP, relative to the number of neurons, compared to the capsaicin control. In the oxytocin-positive control, a significant decrease was observed only at the highest concentration.

[0240] Thus, Granville® rose Zeta-fraction serum inhibits the release of the pro-inflammatory neuropeptide CGRP more effectively than oxytocin.

[0241] Effect of the Zeta-fraction of Granville rose® on the number of TRPV1 receptors

[0242] TRPV1 (transient receptor potential vanilloid 7) receptors belong to the TRP family, a family of receptors sensitive to mechanical, thermal, and certain chemical stimuli. More specifically, neurons possessing TRPV1 receptors are responsible for transmitting pain and itching sensations when stimulated by capsaicin.

[0243] The number of TRPV1 receptors, normalized by the number of sensory neurons, was evaluated after 8 days of treatment with the actives + 30 minutes of treatment with capsaicin, and again 24 hours of treatment with the actives.

[0244] The results are summarized in Tables 4 to 6.

[0245] [Tables4] Treatment Variation / Untreated Control Beta-endorphin 1 pM +3% Zeta-fraction 0.1% -10% Zeta-fraction 0.03% -11%* Zeta-fraction 0.01% = Oxytocin 0.2 mg / mL +7%* Oxytocin 0.06 mg / mL +8%** Oxytocin 0.02 mg / mL +6%

[0246] Under basal conditions, an increase in the quantity of TRPV1 receptors is observed during treatment with the control agents beta-endorphin and oxytocin. Conversely, the trend is a decrease in the quantity of receptors during treatment with Zeta-fraction. The data are significant for the intermediate concentration.

[0247] [Tables5] Treatment Variation / Control untreated Control capsaicin 1OpM +2%

[0248] [Tableauxô] Treatment: active ingredients + CAPSAICIN 10 pM Variation / Capsaicin Control Capsaicin + Beta-endorphin 1 pM -7% Capsaicin + Zeta-fraction 0.1% -11%* Capsaicin + Zeta-fraction 0.03% -4% Capsaicin + Zeta-fraction 0.01% +2% Capsaicin + Oxytocin 0.2 mg / mL -9% Capsaicin + Oxytocin 0.06 mg / mL -5% Capsaicin + Oxytocin 0.02 mg / mL =

[0249] The results show that capsaicin stress does not influence the number of TRPV1 receptors.

[0250] Under these stress conditions, a tendency towards a reduction in the number of TRPV1 receptors is observed for the different conditions evaluated. Only treatment with the Zeta-fraction active ingredient at the highest concentration results in a significant decrease in the number of these receptors.

[0251] Effect of the Zeta-fraction of Granville rose® on cytokine release

[0252] Several pro-inflammatory cytokines were measured after 8 days of treatment with The active ingredients are then treated with capsaicin for 30 minutes, followed by another 6 or 24 hours of treatment with the active ingredients. Values ​​are normalized to the number of sensory neurons.

[0253] The results of the assay are summarized in tables 7 to 9.

[0254] [Tables?] IL-1b IL-6 IL-8 6h 24h 6h 24h 6h 24h Treatment Variation / Control Untreated Beta-endorphin 1 pM -39% -61%* -52%* -14% +7% -21% Zeta-fraction 0.1% -58% -76%** -70%** -57%* -44% -61%* Zeta-fraction 0.03% -63% -82%** -72%** -70%** -47% -68%** Zeta-fraction 0.01% -30% -44% -49%* -39% +7% -8% Oxytocin 0.2 mg / mL -32% -50% -42% -11% -18% -37% Oxytocin 0.06 mg / mL -23% -38% -40% -20% -16% -30% Oxytocin 0.02 mg / mL -10% +19% -8% +18% -6% -20%

[0255] At baseline (in the absence of stress), treatments with beta-endorphin or oxytocin result in variations, generally small or insignificant decreases, in the release of the various cytokines targeted. Conversely, the active ingredient Zeta-fraction at the highest concentrations (0.1% and 0.03%) leads to a significant decrease in the release of the various cytokines IL-1b, IL-6, and IL-8, particularly after an additional 24 hours of treatment.

[0256] [Tables8] IL-1b IL-6 IL-8 6h 24h 6h 24h 6h 24h Treatment Variation / Untreated Control Capsaicin 10 pM +122%* +354%** ** +82%** +178% +236%** * +100%

[0257] [Tables9] IL-1b IL-6 IL-8 6h 24h 6h 24h 6h 24h Treatment: active ingredients + CAPSAICIN 10 pM Variation / Control capsaicin Beta-endorphin 1 pM -38% -69%*** * -58%*** -21% -46%* -3% Zeta-fraction 0.1% -88%** -90%*** * -88%*** * -68% -86%*** * -72% Zeta-fraction 0.03% -82%*** -93%*** * -89%*** * -91%* -78%*** -73% Zeta-fraction 0.01% -56%* -82%*** * -75%*** * -75%* -60%** -41% Oxytocin 0.2 mg / mL -2% -66% -36% -27% -41% -36% Oxytocin 0.06 mg / mL -19% -33% -25% +1% -10% +31% Oxytocin 0.02 mg / mL +38% -4% — +23% +4% +65%

[0258] It is observed that treatment with capsaicin results in a strong, although not always significant, increase in the release of the various cytokines studied.

[0259] Beta-endorphin control often significantly induces a reduction in the release of interleukins IL-1b, IL-6 and IL-8. Regarding oxytocin, the results observed are generally not significant.

[0260] For its part, treatment with the active ingredient Zeta-fraction has the effect of strongly and significantly reducing the release of interleukins at both times tested.

[0261] Granville® Rose Zeta-fraction serum inhibits the release of pro-inflammatory cytokines more effectively than oxytocin.

[0262] Effect of the Zeta-fraction of Granville® rose on the survival of sensory neurons

[0263] The survival of sensory neurons was evaluated after 8 days of treatment with the active ingredients + 30 minutes of treatment with capsaicin, and again 24 hours of treatment with the active ingredients.

[0264] The results are summarized in Tables 10 to 12.

[0265] [TableauxlO] Treatment Variation / Control Untreated Beta-endorphin 1 pM +16% Zeta-fraction 0.1% +112%*** Zeta-fraction 0.03% +161%**** Zeta-fraction 0.01% +66% Oxytocin 0.2 mg / mL +63% Oxytocin 0.06 mg / mL +48% Oxytocin 0.02 mg / mL +13%

[0266] Under basal conditions, a strong and significant increase in the survival of sensory neurons is observed with the active Zeta-fraction, mainly at the two highest concentrations.

[0267] [Tableauxll] Treatment Variation / Untreated Control Control Capsaicin lOpM -66%*

[0268] [Tables 12] Treatment: active ingredients + CAPSAICIN 10 pM Variation / Capsaicin Control Capsaicin + Beta-endorphin 1 pM +79% Capsaicin + Zeta-fraction 0.1% +466%**** Capsaicin + Zeta-fraction 0.03% +478%**** Capsaicin + Zeta-fraction 0.01% +231%** Capsaicin + Oxytocin 0.2 mg / mL +65% Capsaicin + Oxytocin 0.06 mg / mL = Capsaicin + Oxytocin 0.02 mg / mL -1%

[0269] As expected, an increase in cell death is observed in the presence of capsaicin.

[0270] Under these stress conditions, treatment with the active ingredient Zeta-fraction strongly promotes the survival of sensory neurons. A trend towards increased cell survival is observed with beta-endorphin and oxytocin at the highest concentration.

[0271] -> Granville rose® serum (Zeta-fraction) promotes neuronal survival sensory receptors more effectively than oxytocin.

[0272] Effect of the Zeta-fraction of Granville® rose on the length of extensions sensory neurons

[0273] The length of the sensory neuron extensions was assessed after 8 days of treatment with the actives + 30 minutes of treatment with capsaicin, and again 24 hours of treatment with the actives.

[0274] The results are summarized in Tables 13 to 15.

[0275] [Tables 13] Treatment Variation / Untreated Control Beta-endorphin 1 pM +80% Zeta-fraction 0.1% +177%** Zeta-fraction 0.03% +260%*** Zeta-fraction 0.01% +77% Oxytocin 0.2 mg / mL +56% Oxytocin 0.06 mg / mL +116%* Oxytocin 0.02 mg / mL +17%

[0276] Under basal conditions, a strong and significant increase in the length of sensory neuron processes is observed with the active ingredient Zeta-fraction at the highest concentrations, as well as with oxytocin at the intermediate concentration. Regarding the other conditions, a tendency towards an increase in process length is observed, often quite marked, although without reaching statistical significance.

[0277] [Tables 14] Treatment Variation / Untreated Control Control Capsaicin lOpM -67%*

[0278] [Tables 15] Treatment: Active ingredients + CAPSAICIN 10 pM Variation / Capsaicin Control Capsaicin + Beta-endorphin 1 pM 158% Capsaicin + Zeta-fraction 0.1% +486%**** Capsaicin + Zeta-fraction 0.03% +554%**** Capsaicin + Zeta-fraction 0.01% +195% Capsaicin + Oxytocin 0.2 mg / mL +50% Capsaicin + Oxytocin 0.06 mg / mL 73% Capsaicin + Oxytocin 0.02 mg / mL +43%

[0279] As expected, in the presence of capsaicin, a decrease in the length of the processes of sensory neurons is observed.

[0280] Under these stressful conditions, treatment with the active ingredient Zeta-fraction significantly and strongly increases the length of sensory neuron extensions. A similar trend is observed with treatment using beta-endorphin or oxytocin; however, the values ​​obtained do not reach statistical significance.

[0281] -> The effect of the Zeta-fraction on the increase in the total length of the pro The lengthening of sensory neurons is even more pronounced under stressful conditions. Conclusions

[0282] The various active ingredients tested (oxytocin and Zeta-fraction of Granville rose®), as well as capsaicin and beta-endorphin, all have a significant impact on the survival of sensory neurons. Thus, depending on the treatment conditions, the number of sensory neurons can vary considerably. For example, three times more neurons survive under basal conditions (without stress) compared to the capsaicin stress condition. To ensure that these variations do not affect the analysis of the results (such as the expression of the different receptors, the amount of CGRP released, or cytokines induced), these parameters were normalized by the number of sensory neurons observed.

[0283] The Zeta-fraction of Granville® rose has a major positive impact on the survival of sensory neurons whether in basal or stress conditions.

[0284] Under basal conditions, it moderately inhibits the expression of TRPV1 receptors but does not modulate CGRP release. However, it induces an anti-inflammatory effect with a decrease in the release of pro-inflammatory cytokines.

[0285] Under capsaicin stress conditions, the Zeta-fraction of Granville rose® moderately inhibits the expression of TRPV1 receptors. It also significantly inhibits It specifically inhibits the release of the neuropeptide CGRP and pro-inflammatory cytokines. Furthermore, it helps preserve the survival of sensory neurons and protect their neuronal extensions.

[0286] In conclusion, these experiments show that the Zeta-fraction of Granville rose® possesses protective and soothing properties at the three concentrations tested: 0.1%, 0.03%, and 0.01%. The observed effect is generally greater with the active ingredient according to the invention (Zeta-fraction of Granville rose®) than with oxytocin. Thus, the Zeta-fraction of Granville rose® according to the invention has a neuroprotective and / or neuro-soothing effect on the skin and / or lips and can prevent and / or reduce irritation and / or micro-inflammation. Example 2: Cosmetic formulations

[0287] Composition in the form of an aqueous facial gel

[0288] [Tableauxlô] Cosmetic Ingredients: Zeta-fraction (bioactive fraction isolated from rose petals according to the invention) * 50.00% Preservatives 0.50% Sugar 0.20% Carbomer 0.70% Purified water q.s. 100% Tetrasodium EDTA powder 0.20% Sodium hydroxide 0.17%

[0289] * as described above

[0290] The isolated bioactive fraction of rose petals according to the invention and the preservatives are mixed and homogenized at room temperature with stirring. The sugars are added with stirring, then the carbomer, then the water and EDTA with stirring. The aqueous gel is then neutralized by adding sodium hydroxide with stirring until a homogeneous gel is obtained.

[0291] Applied to the skin of the face, this aqueous gel gives a destressing and soothing effect to the skin.

[0292] Composition in the form of a micronutrient salt-serum for the face

[0293] [Tables 17] Cosmetic Ingredients Content Purified Water Qsp 100.00% Glycols 13.0% Preservatives 0.60% Carbomer 0.80% Glyceryl Stearate Citrate 0.70% Lecithin and Sodium Acrylates Copolymer (Lecigel PCR Negative) 1.20% Isostearyl Isostearate 9.0% Bis-Diglyceryl Polyacyladipate-2 (Softisan 649 MB) 1.0% Silica 2.0% Nacres 1.0% Rose Cryoextract* 3.0% Centella Asiatica Extract 0.5% Horse Chestnut Extract 0.1% Zeta-fraction (bioactive fraction isolated from rose petals according to the invention)* 0.3% Tocopheryl Acetate 0.10%

[0294] * as described above

[0295] The aqueous phase ingredients (water, glycols, and carbomer) are mixed at 80°C with stirring. Preservatives are then added, followed by gelling agents at 80°C with stirring, and the temperature is then lowered to 75°C. Surfactants are then emulsified in the aqueous phase with stirring. The fillers and pearlescent pigments, previously mixed and homogenized, are added at 40°C. The cryo-extract, the isolated bioactive fraction of rose petals according to the invention, and the other extracts are then added at 40°C, with stirring until the mixture reaches 30°C.

[0296] After application to the face, signs of micro-inflammation are reduced, the skin is soothed.

Claims

Demands

1. Non-therapeutic cosmetic use of a rose extract in the form of an isolated bioactive serum fraction, obtained from fresh roses of the Evanrat variety, to soothe healthy skin and / or healthy lips, characterized in that said extract is obtained by a process comprising the steps: a) cleaning of the plant material, maceration, pressing and then mechanical separation of the plant material to obtain an intracellular colloidal dispersion (ICD) and a fiber-enriched material (fraction A); b) destabilization of the ICD with electromagnetic waves and then mechanical separation of the intracellular colloidal dispersion (ICD) to obtain supernatant A and a Membrane Fraction (fraction B); c) adjustment of the pH level in supernatant A until a pH greater than 6 is obtained, preferably a pH ranging from 6 to 7, and then mechanical separation of supernatant A to obtain supernatant B and fraction C (cytoplasmic fraction);d) Adjustment of the pH level in supernatant B until a pH below 4.5 is obtained, then mechanical separation of supernatant B to give a "Bioactive Serum Fraction" and a fraction D (precipitate); and e) Optionally, mixing of the "Bioactive Serum Fraction" with at least one preservative and / or stabilizer.

2. Non-therapeutic cosmetic use according to claim 1, characterized in that the rose extract in the form of an isolated bioactive serum fraction is obtained from fresh rose petals of the Evanrat variety.

3. Non-therapeutic cosmetic use according to claim 1, characterized in that the isolated bioactive serum fraction comprises sugars, polyphenols and minerals.

4. Non-therapeutic cosmetic use according to any one of claims 1 to 3, to prevent and / or reduce non-pathological phenomena of irritation of healthy skin and / or healthy lips.

5. Use according to any one of claims 1 to 4, characterized in that said non-pathological irritation phenomena are induced in particular by at least one condition selected from air pollution, stress, irritant molecules, thermal shocks, and / or friction.

6. Use according to any one of claims 1 to 5, characterized in that said extract is present in a cosmetic composition in a content ranging from 0.001% to 50%, in particular from 0.01% to 20%, preferably from 0.01% to 10% and preferably still from 0.011% to 5% by weight of raw material in relation to the total weight of said cosmetic composition.

7. Use according to claim 6, characterized in that said cosmetic composition further comprises at least one cosmetic adjuvant selected from antioxidants, perfumes, vitamins, thickening agents, emollients, moisturizing agents, anti-aging agents, lifting agents, tightening agents, plumping agents, soothing agents, anti-pollution agents, brightening or depigmenting agents, fillers, mother-of-pearl and mixtures thereof.

8. Use according to claim 6 or claim 7, characterized in that said cosmetic composition 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 especially for the skin, a balm, a gloss or a stick especially for the lips.

9. Cosmetic use according to any one of claims 1 to 8 characterized in that said extract or cosmetic composition is intended for application to subjects with irritated and / or stressed but healthy skin and / or lips.

10. Cosmetic use according to any one of claims 1 to 9, characterized in that said extract or cosmetic composition is intended for topical application on healthy skin and / or healthy lips and in particular on the skin of the face and / or neck.

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