METHOD FOR THE VALORIZATION OF CANANGA ODORATA PLANT WASTE AND CANANGA ODORATA EXTRACT

Abstract

A process for valorizing Cananga Odorata waste to obtain a Cananga Odorata extract, said process comprising a first solid / liquid extraction step carried out on said plant waste using an extraction solvent, followed by a second solid / liquid separation step and a third recovery step of the liquid phase constituting the extract, the extraction solvent being anhydrous and non-polar. Use of said extract for the cosmetic treatment of the skin and / or mucous membranes and cosmetic and / or dermatological compositions comprising said extract.

Description

Title of the invention: METHOD FOR THE VALORIZATION OF A PLANT WASTE FROM CANANGA ODORATA AND CANANGA ODORATA EXTRACT technical field

[0001] The invention relates to a process for valorizing Cananga odorata waste to obtain an extract of Cananga odorata. It also relates to an extract of Cananga odorata, the use of said extract for the cosmetic treatment of the skin and / or mucous membranes, as well as a cosmetic and / or dermatological composition comprising said extract. Previous technique

[0002] Life on Earth follows a 24-hour rhythm largely determined by the daily oscillations of light, due to the Earth's axial rotation. It is now well established that our bodies respond to day / night cycles known as circadian rhythms. This multi-oscillatory network allows our bodies to adapt to environmental changes and adjust our internal clocks accordingly.

[0003] The central clock that regulates the 24-hour cycle throughout our body is located in the hypothalamus in the brain. This area of ​​the brain responds to light received through the retina of the eye. The central clock communicates this synchronizing information to peripheral organs and tissues via neural and hormonal signals. Similar, so-called peripheral clocks have been discovered within the cells of almost all tissues, including skin.

[0004] Indeed, skin cells, particularly keratinocytes, melanocytes, and fibroblasts, also possess an independent circadian clock system, which consequently regulates the overall homeostasis of human skin tissue. These peripheral clocks function in the same way as the central clock, by regulating cell activity through the expression of clock genes that control and synchronize molecular and cellular mechanisms according to the day / night cycle. Thus, our skin experiences different phases in rhythm with the day / night cycle: the activation of these clock genes allows the skin to prepare for, react to, and adapt to daily changes in the external environment.

[0005] The skin is naturally exposed to diurnal changes, including temperature, light, humidity, and UV radiation, due to its role as an interface between the body and the external environment. The skin's circadian clock responds to daily fluctuations and adjusts the periodicity of skin functions: hydration and transepidermal water loss (TEWL). trans-epidermal water), keratinocyte proliferation rate, capillary blood flow, sebum production, temperature, surface pH and facial wrinkles in humans.

[0006] In the skin, clock genes control and synchronize cellular activities such as DNA damage repair, cell proliferation, cell cycle, apoptosis, and metabolism.

[0007] This chronobiological phenomenon shows that during the day, most of the cellular energy is used to protect the skin, while during the night, most of the energy is used to restore, repair, regenerate, and prepare skin cells. Cellular activity is much greater during the night, with cells multiplying intensely and a higher metabolism (peak cell division around 1 a.m.).

[0008] Cellular aging processes are closely linked to an imbalance in the pro / antioxidant balance that develops gradually with age in relation to a dysfunction of circadian clocks. Several studies have shown differences in protein oxidation at different times of day, indicating altered sensitivity to oxidative stress depending on the circadian clock. The circadian clock plays a vital role in maintaining ROS (Reactive Oxygen Species) at normal levels and protecting cells and tissues from oxidative damage. In particular, the central molecular clock protein Bmall not only regulates the circadian rhythm but also plays a role in redox homeostasis and enhancing cell survival under oxidative conditions. There is evidence of a role for Bmall in regulating ROS in several tissue types (pancreas, brain, skin).The overall suppression of Bmall produces an accelerated aging phenotype, which is a consequence of increased oxidative stress.

[0009] Given the potentially harmful nature of ROS, antioxidants are produced to inhibit the oxidation of biological molecules and balance the oxidative state of cells. The transcription factor Nrf2 plays a major role in regulating the expression of antioxidant proteins, protecting cells against oxidative damage triggered by injury or inflammation. Under normal conditions, Nrf2 is tightly bound to Keapl in the cytoplasm and has a very short lifespan of 10 to 30 minutes (degradation by the proteasome). Under stress, ROS levels increase, Nrf2 dissociates from Keapl and translocates to the nucleus. In the nucleus, Nrf2 binds to the ARE (Antioxidant Response Element) and thus controls antioxidant capacity.

[0010] The Nrf2 antioxidant pathway is regulated by the circadian clock. Several studies demonstrate the direct role of Bmall in the regulation of Nrf2 to control the Antioxidant and anti-inflammatory responses. Indeed, Bmall binds to the Nrf2 / ARE promoter via the E-Box element, thereby regulating the rhythmic activation of the Nrf2 transcription factor. Bmall suppression decreases the Nrf2 response and leads to ROS accumulation and increased production of pro-inflammatory cytokines. The interaction of circadian components with antioxidant pathways via Nrf2 plays a crucial role in many ROS-induced diseases. Conversely, cells overexpressing Bmall have shown increased levels of mRNA for the antioxidant enzymes NQO1, SOD, GPX, and Nrf2, and this expression is closely linked to a decrease in ROS. This interconnected loop between Bmall and Nrf2 has also been demonstrated in the epidermis, particularly in a culture of human keratinocytes exposed to ozone.

[0011] Thus, we have seen that day / night cycles, called circadian rhythms, allow our body to adapt to environmental changes and adjust our internal clocks accordingly. These clocks are present in almost all tissues, including skin tissue. They regulate cell activity through the expression of clock genes that control and synchronize molecular and cellular mechanisms according to the day / night cycles. During cellular aging, an imbalance in the pro- and antioxidant balance develops in relation to a dysfunction of the circadian clock. The global suppression of the central molecular clock protein Bmall reduces the response of the Nrf2 antioxidant pathway, which is a consequence of increased oxidative stress and an accelerated aging phenotype.

[0012] Skin protection mechanisms are dysregulated during intrinsic and extrinsic aging. These protective mechanisms are primarily activated at night in parallel with the cell's biological clock (activation of the "night clock" genes BMAL1 and CLOCK). Thanks to the activation of these nocturnal protective mechanisms, such as the stimulation of the Nrf2 antioxidant pathway and the activation of transcription of genes encoding antioxidant enzymes, the cell is able to cope with daytime environmental stress, namely photopollution, to protect its proteome from oxidation and carbonylation. Therefore, it is important to act both day and night for optimal protection of skin tissue.

[0013] A first problem that the invention proposes to solve is to provide an active ingredient that stimulates the nocturnal and diurnal mechanisms of skin protection against aggressions and skin aging.

[0014] There are many active ingredients that can protect the skin, including products derived from the Ylang-Ylang flower (Cananga odorata) which are particularly known for their beneficial effect against skin aging.

[0015] Ylang-Ylang (Cananga odorata), whose name means "flower of flowers", is a species belonging to the Annonaceae family, native to the humid forests of Southeast Asia.

[0016] This is a fast-growing tree, reaching 25 to 30 meters in height in its wild form, in its native habitat. Its bark is gray, and its evergreen leaves are alternate, lanceolate, and without stipules, and are bright green. Its flowers, axillary or grouped in cymes, are composed of three valvate sepals and six drooping, strap-like petals. They are initially white, then turn green, and finally become yellow. Their base turns red when the flower matures, at which point it is very fragrant. Flowering occurs throughout the year, but is particularly abundant during the warm and humid season (December to April / May). The fruits, composed of ovoid mericarps, have black carpels.

[0017] Ylang-ylang is now mainly cultivated in the islands of the Indian Ocean (Comoros, Madagascar, Mayotte) for its flowers, which contain an essential oil highly prized in the perfume, health, and cosmetics industries. Once mature, the flowers are harvested by hand between dusk and dawn, when the flower's fragrance is at its peak.

[0018] Ylang-ylang is a plant widely used throughout the world in the form of essential oil extracted from its flowers. This essential oil is obtained by hydrodistillation or, more commonly, steam distillation of freshly picked flowers. The plant residues resulting from the hydrodistillation of the flowers are called press cake and constitute waste for the industry.

[0019] Ylang-ylang essential oil is well known for its cosmetic applications. It is primarily the essential oils derived from the flowers of Cananga odorata that are described for use on skin and hair.

[0020] These essential oils of Cananga odorata flowers contain more than a hundred volatile compounds belonging to several phytochemical families including sesquiterpenes such as germacrene D, beta-caryophyllene and alpha-farnesene, esters such as methyl benzoate, benzyl benzoate, benzyl acetate and geranyl acetate, monoterpenols such as linalool and phenylpropanoid compounds such as p-methylanisole.

[0021] However, among the volatile compounds constituting these essential oils of Cananga odorata, a significant number are allergens. And the negative effects of Ylang-Ylang essential oil are widely known and described in the literature as potentially causing skin irritation and hyperpigmentation.

[0022] Several compounds identified in Ylang-Ylang flower essential oil are on the list of molecules classified as "allergens" by the regulations European regulation concerning cosmetic products (Commission Regulation (EU) 2023 / 1545 of 26 July 2023 amending Regulation (EC) No 1223 / 2009 of the European Parliament and of the Council as regards the labelling of allergenic perfume substances in cosmetic products).

[0023] Essential oil manufacturers communicate the following allergen contents: Linalool: 2 to 16%; Benzyl benzoate: 5 to 12%; Geraniol: 0.2 to 5%; Famesol: 1.5 to 5%; Benzyl salicylate: 1 to 4%; Eugenol: 0.1 to 0.9%; Benzyl alcohol: 0.1 to 0.5%; Isoeugenol: 0.1 to 3.0%.

[0024] The total content of allergenic compounds in the essential oils of Cananga odorata flower can therefore vary between, approximately, 10% and 46% of the overall composition.

[0025] Document FR 3 110 416 describes a process for obtaining Ylang-Ylang concrete and absolute, comprising numerous steps, some of which are very energy-intensive, such as the removal of the organic solvent from the liquid phase to obtain a concrete, and the removal of waxes present in the concrete to obtain an absolute. The extraction solvents used, notably hexane and ethyl acetate, have the disadvantage of toxicity and environmental pollution. Finally, and most importantly, another significant issue is the presence in the products obtained of a large quantity of compounds classified as allergens, with concentrations ranging from 36% to 67%.

[0026] A second problem that the invention aims to solve is to provide an active ingredient that reduces or eliminates allergenic risks, and is therefore very low in allergenic compounds.

[0027] In parallel with this issue, the applicant is particularly concerned and sensitive to the environmental issues related to the development and creation of new products, and pays particular attention to eco-design.

[0028] Eco-design is defined by ADEME (French Agency for Environment and Energy Management) as a preventive and innovative approach which makes it possible to reduce the negative impacts of a product on the environment over its entire life cycle (LCA for Life Cycle Analysis), while maintaining its qualities of use.

[0029] The invention therefore also falls within a context of reducing the environmental footprint of industrial and consumption activities.

[0030] According to the European Union, up to 80% of the environmental impacts of a product, device, or process are linked to decisions made during the design phase. Eco-design is therefore a major challenge in reducing the negative impacts of human production and consumption on the environment.

[0031] In other words, the problem that the invention aims to solve is that of providing an effective active ingredient to stimulate the nocturnal and diurnal mechanisms of skin protection against aggressions and skin aging, while reducing or eliminating the allergenic risks associated with the use of the active ingredient, and attempting to offer a more eco-responsible solution. Description of the invention

[0032] The applicant found, quite surprisingly, that it was possible to obtain an extract of Cananga odorata that met the needs mentioned above by valorizing a plant waste of Cananga odorata.

[0033] According to a first aspect, the invention relates to a process for valorizing a plant waste of Cananga odorata to obtain an extract of Cananga odorata, said process comprising a first solid / liquid extraction step carried out on said plant waste using an extraction solvent, followed by a second solid / liquid separation step and a third recovery step of the liquid phase constituting the extract.

[0034] The invention is characterized in that the extraction solvent is anhydrous and nonpolar. Preferably, the extraction solvent is miscible with water. By "anhydrous," we mean an extraction solvent containing no added water, and only traces of water with a content of less than 1% by weight. By "nonpolar," we mean an extraction solvent whose resulting dipole moment is zero: either because the solvent contains no polar groups, or because the solvent contains polar groups whose geometry causes the dipole moment to vanish. By "miscible with water," we mean that the extraction solvent has the capacity to form a homogeneous mixture with water.

[0035] In another aspect, the invention relates to an extract of Cananga odorata obtained according to the process of the invention. The invention also relates to an extract of Cananga odorata characterized in that it comprises an allergenic compound content of less than 0.1% by weight relative to the total weight of the extract, the allergenic compounds being defined according to EU Regulation 2023 / 1545.

[0036] According to another aspect, the invention relates to the non-therapeutic use of the extract of Cananga odorata according to the invention, or of a composition comprising it, for the cosmetic treatment of the skin and / or mucous membranes.

[0037] Finally, according to another aspect, the invention relates to a cosmetic and / or dermatological composition comprising at least the extract of Cananga odorata according to the invention.

[0038] The invention is characterized by the valorization of a plant waste product considered as an "ultimate" waste product in the extraction of molecules of interest from Cananga odorata, particularly in processes for obtaining Cananga odorata essential oil. In other words, the raw material on which the process according to the invention is carried out is not the fresh or dried Ylang-Ylang (Cananga odorata) flower, but a Cananga odorata plant waste product resulting from an extraction process, preferably an extraction process for obtaining Cananga odorata essential oil. The plant residues resulting from the hydrodistillation or steam distillation of Cananga odorata flowers to obtain Cananga odorata essential oil are called press cakes and constitute a waste product for this sector.

[0039] The term “Cananga odorata plant waste” means a fresh or dried, whole or ground press cake obtained from fresh or dried Ylang Ylang flowers, advantageously a ground dry press cake obtained from fresh flowers. The Cananga odorata flower press cake is preferably the waste product from the Cananga odorata essential oil extraction process.

[0040] Contrary to expectations, it is by using a waste product considered "ultimate" in the manufacture of Cananga odorata essential oil that, thanks to the specific process of the invention, an effective active ingredient can be obtained to stimulate the nocturnal and diurnal mechanisms of skin protection against aggressions and skin aging, while reducing or eliminating the allergenic risks associated with the use of the active ingredient.

[0041] Advantageously, the extraction solvent comprises at least one diol comprising 3 or 4 carbon atoms, preferably at least 10% by weight relative to the total weight of the extraction solvent.

[0042] In a particular and preferred embodiment, the extraction solvent is either made up of a diol comprising 3 or 4 carbon atoms, or made up of a mixture of betaine, glycerin and a diol comprising 3 or 4 carbon atoms, or made up of a mixture of sorbitol, a diol comprising 3 or 4 carbon atoms and glycerin.

[0043] Preferably, the extraction solvent is either made up of propanediol and / or butanediol, or made up of a mixture of betaine, propanediol and / or butanediol and glycerin, or made up of a mixture of sorbitol, propanediol and / or butanediol and glycerin.

[0044] Preferably, the extraction solvent may consist solely of propanediol or solely of butanediol, or a mixture of the two. Preferably, the extraction solvent may consist of a mixture of betaine, glycerin, and propanediol, or of a mixture of betaine, glycerin, and of butanediol, or be composed of a mixture of betaine, glycerin, propanediol and butanediol. Preferably, the extraction solvent may be composed of a mixture of sorbitol, glycerin and propanediol, or be composed of a mixture of sorbitol, glycerin and butanediol, or be composed of a mixture of sorbitol, glycerin, propanediol and butanediol.

[0045] Propanediol is preferably propane-1,3-diol or propane-1,2-diol according to the IUPAC (International Union of Pure and Applied Chemistry) nomenclature, and more preferably propane-1,3-diol. Propane-1,2-diol is also called propylene glycol. These solvents belong to the glycol family. They can be obtained from sugars of vegetable origin and from processes for transforming these sugars listed and approved by the COSMOS standard.

[0046] Butanediol is preferably butane-1,3-diol according to IUPAC nomenclature, also known as butylene glycol. Similarly, it can be obtained from sugars of vegetable origin and from processes for transforming these sugars listed and approved by the COSMOS standard.

[0047] Certification according to the international COSMOS standard makes it possible to label natural or organic products by studying all aspects of the supply, manufacture, marketing and control of cosmetic products.

[0048] Betaine refers to 2-(trimethylazaniumyl)acetate according to IUPAC nomenclature (syn.: trimethylglycine, betaine glycine), which belongs to the family of quaternary ammonium compounds derived from amino acids. This betaine may be available in anhydrous or hydrated form. It is preferably in anhydrous form. It can be obtained from plant material and processing methods listed and approved by the COSMOS standard. It is generally extracted from sugar beet.

[0049] Sorbitol is defined as (2R,3R,4R,5S)-hexane-1,2,3,4,5,6-hexol according to IUPAC nomenclature, belonging to the polyol family. It can be obtained from plant material and processing methods listed and approved by the COSMOS standard.

[0050] Glycerin is defined as propane-1,2,3-triol according to IUPAC nomenclature, belonging to the glycol family. It can be obtained from vegetable matter and transformation processes listed and approved by the COSMOS standard.

[0051] The invention thus offers the advantage, thanks to these extraction solvents, of providing an eco-design benefit because said solvents are "agro-solvents," that is to say, obtained from plant matter, therefore from renewable materials. Preferably, the extraction solvents are produced using a process approved by the COSMOS standard, which is environmentally friendly.

[0052] Another advantage of the extract according to the invention is that it can be used directly by the formulator for the preparation of cosmetic and / or dermatological compositions, without requiring prior removal of the solvent from the extract. Indeed, these extraction solvents are so-called "ingredient" solvents, that is to say, solvents that can be used in the composition of said cosmetic and / or dermatological compositions. They are compatible and non-toxic to humans and the environment.

[0053] Another advantage of the invention is that it provides an extract with a reduced water content, thus avoiding the need to add additives or preservatives to stabilize the extract's shelf life. By "reduced water content extract," we mean an extract obtained by extraction from the fresh or dried plant using an anhydrous solvent, as defined above, said extract being characterized by a residual water content advantageously less than 20% by weight, preferably less than 5%.

[0054] Furthermore, when the components of this solvent are present in certain molar ratios, the solvent possesses the properties of a NaDES (Natural Deep Eutectic Solvent), that is, a eutectic solvent composed of molecules found in nature and linked together by intermolecular interactions, in particular hydrogen bonds. The solvent is notably liquid at room temperature, which facilitates the implementation of the process leading to the extract of the invention. The extraction solvent according to the invention is preferably a NaDES solvent.

[0055] In the case where the extraction solvent consists of a mixture of betaine, glycerin and diol comprising 3 to 4 carbon atoms, for example propanediol (mixture noted in this case BGP), the molar proportions between the different substances are preferably those set out below.

[0056] The molar proportions between betaine and diol comprising 3 to 4 carbon atoms are preferably between 1:0.5 and 1:1.4, or between 1:3.5 and 1:7, more preferably between 1:4 and 1:6, even more preferably 1:5.

[0057] The molar proportions between betaine and glycerin are preferably between 1:0.5 and 1:1.4, or between 1:3.5 and 1:7, more preferably between 1:0.7 and 1:1.2, even more preferably 1:1.

[0058] The molar proportions between glycerin and diol comprising 3 to 4 carbon atoms are preferably between 1:7 and 7:1, more preferably between 1:5 and 5:1, even more preferably 1:5.

[0059] The molar proportions of betaine, glycerin, and the diol comprising 3 to 4 carbon atoms are preferably between 1:1:3 and 1:1:7, even more preferably between 1:1:4 and 1:1:6, and even more preferably about 1:1:5. In a particularly preferred mode, the extraction solvent is a betaine NaDES / glycerin / diol comprising 3 to 4 carbon atoms having these molar proportions, and particularly that having a molar ratio 1:1:5.

[0060] In the case where the extraction solvent consists of a mixture of sorbitol, a diol comprising 3 to 4 carbon atoms, for example propanediol, and glycerin (mixture noted in this case SPG), the molar proportions between the different substances are preferably those shown below.

[0061] The molar proportions between sorbitol and diol comprising 3 to 4 carbon atoms are preferably between 1:0.5 and 1:7, more preferably between 1:0.7 and 1:3.5, more preferably between 1:0.8 and 1:1.4, and even more preferably 1:1.

[0062] The molar proportions between sorbitol and glycerin are preferably between 1:7 and 1:0.5, more preferably between 1:6 and 1:1.5, more preferably between 1:5.5 and 1:3.5, even more preferably 1:5.

[0063] The molar proportions between the diol comprising 3 to 4 carbon atoms and glycerin are preferably between 1:1 and 1:7, more preferably between 1:4 and 1:6, even more preferably 1:5.

[0064] The molar proportions between sorbitol, the diol comprising 3 to 4 carbon atoms and glycerin are preferably between 1:1:3 and 1:1:7, even more preferably 1:1:4 and 1:1:6, and even more preferably about 1:1:5. In a particularly preferred mode, the extraction solvent is a NaDES sorbitol / diol comprising 3 to 4 carbon atoms / glycerin having these molar proportions, and particularly one having a molar ratio of 1:1:5.

[0065] In practice, the solvent used can be produced by mixing either betaine, glycerin, and a diol comprising 3 to 4 carbon atoms, or sorbitol, glycerin, and a diol comprising 3 to 4 carbon atoms, in a stirred reactor, until a homogeneous, clear, colorless liquid mixture is obtained. This mixture can be prepared at a temperature between 2°C and 100°C and for 0.5 to 6 hours, preferably at 40°C to 70°C for 1 to 2 hours.

[0066] The extraction solvent is preferably free of hexane and ethyl acetate.

[0067] The process of valorizing a plant waste of Cananga odorata to obtain an extract of Cananga odorata involves a solid / liquid extraction step.

[0068] Solid / liquid extraction can be carried out by various techniques well known to those skilled in the art, such as maceration, remaceration, digestion, dynamic maceration, decoction, fluid bed extraction, microwave-assisted extraction, ultrasonic-assisted extraction, counter-current extraction, percolation, repercolation, leaching, reduced-pressure extraction, diacolation.

[0069] In practice, the plant / solvent mass ratio applied for the extraction step is between 1 / 99 and 25 / 75, advantageously between 3 / 97 and 15 / 85, preferably between 5 / 95 and 10 / 90. The extraction step is preferably carried out at a temperature between 2°C and 100°C, more preferably between 20°C and 80°C. The extraction step can be maintained for a period from a few minutes to several days.

[0070] In order to optimize the extraction of active compounds while protecting these compounds from oxidation by atmospheric oxygen, the solid / liquid extraction step is advantageously carried out under agitation and / or under a nitrogen atmosphere.

[0071] According to the invention, the solid / liquid extraction step is followed by a solid / liquid separation step, the objective being to recover the liquid phase, also called the solid / liquid separation filtrate, containing the active material. This separation can be carried out by any technique known to those skilled in the art, in particular draining, pressing, dewatering, centrifugation, or filtration.

[0072] Optionally, the liquid / solid separation step may be followed by a concentration and / or fractionation step, which yields a concentrate or fraction in liquid or semi-solid form, depending on the concentration factor. In practice, the concentration or fractionation step may be carried out by evaporation under reduced pressure, ultrafiltration, nanofiltration, reverse osmosis, or chromatography.

[0073] Preferably, the solid / liquid separation filtrate or concentrate further undergoes one or more clarification steps. To carry out this clarification step, a person skilled in the art may use any type of filtration known in the relevant field.

[0074] Finally, for the purpose of packaging, the process for obtaining the extract according to the invention may include a sterilizing filtration. Sterilizing filtration is conventionally carried out by filtering the product through a filter comprising pores with a diameter of 0.22 µm. Preferably, the sterilizing filtration step is the final step of the process.

[0075] The invention also relates to an extract of Cananga odorata obtained according to the process of the invention. The process of the invention makes it possible to obtain an extract of Cananga odorata that reduces or eliminates allergenic risks, thus offering the cosmetics industry an active ingredient perfectly in line with the expectations of this field.

[0076] Unlike the prior art extracts of Cananga odorata which include large quantities of allergenic compounds, generally between 10% and 46% by weight for the essential oil, and between 36% and 67% by weight for the extracts according to document FR3110416, the extract according to the invention includes only a very small quantity of allergenic compounds.

[0077] According to another aspect, the invention relates to an extract of Cananga odorata comprising an allergenic compound content of less than 0.1% by weight compared to the total weight of the extract, the allergenic compounds being defined according to EU Regulation 2023 / 1545. The Cananga odorata extract according to the invention is devoid of allergenic effect.

[0078] By allergenic compounds, we mean the molecules listed and classified as "allergens" by European regulations concerning cosmetic products: Commission Regulation (EU) 2023 / 1545 of 26 July 2023 amending Regulation (EC) No 1223 / 2009 of the European Parliament and of the Council as regards the labelling of allergenic perfume substances in cosmetic products.

[0079] According to the invention, the Cananga odorata extract preferably comprises a benzyl benzoate content of less than 0.05% by weight relative to the total weight of the extract, and / or a benzyl salicylate content of less than 0.05% by weight relative to the total weight of the extract.

[0080] One of the advantages of the invention therefore lies in the fact that the Cananga odorata extract of the invention can be used directly by the formulator for the preparation of cosmetic and / or dermatological compositions, without requiring prior removal of the extraction solvent. The extraction solvent is considered an "ingredient solvent".

[0081] According to the invention, the Cananga odorata extract is preferably non-oily. It preferably has a reduced water content, that is, a water content of less than 20% by weight, preferably less than 5% by weight. It is preferably non-polar. The Cananga odorata extract is most preferably non-oily and contains less than 20% by weight of water, and more preferably non-oily, non-polar, and contains less than 20% by weight of water. By "non-oily," it is understood that the extract does not contain oil, or traces of oil with a content of less than 1% by weight.

[0082] According to another aspect, the invention relates to the non-therapeutic use of the extract of Cananga odorata according to the invention, or of a composition comprising it, for the cosmetic treatment of the skin and / or mucous membranes.

[0083] The Applicant has further discovered that the extract of the invention makes it possible to stimulate the nocturnal and diurnal mechanisms of protection of the skin against aggressions and skin aging.

[0084] The extract according to the invention has particularly interesting biological properties, in particular for improving the appearance of the skin and / or mucous membranes, for improving skin resistance and / or elasticity, and / or for the treatment or prevention of skin aging, wrinkles, or skin laxity.

[0085] The invention relates to the non-therapeutic use of the extract of Cananga odorata according to the invention, or of a composition comprising it, to combat skin aging and / or protect the skin from external aggressions.

[0086] The invention relates to the non-therapeutic use of the extract of Cananga odorata according to the invention, or of a composition comprising it, to increase skin hydration, and / or strengthen the skin barrier function, and / or improve skin microrelief, and / or improve skin radiance and complexion.

[0087] The invention relates to the non-therapeutic use of the Cananga odorata extract according to the invention, or of a composition comprising it, to increase the transcriptomic expression of antioxidant enzymes and / or to stimulate the Nrf2 antioxidant pathway, and / or to increase protection against stress-induced protein carbonylation in skin tissue.

[0088] Indeed, as detailed in the experimental part, the Cananga odorata extract according to the invention has a biological efficacy on the activation of the Nrf2 antioxidant pathway and on the increase of the transcriptomic expression of antioxidant genes, in cultures of keratinocytes that are not synchronized and are synchronized in "night" mode, i.e. day and night.

[0089] The demonstrated efficiencies are respectively as follows: - Increased expression of the Nrf2 transcription factor in the nucleus of unsynchronized and synchronized keratinocytes in "night" mode. - Increased transcriptomic expression of the oxidation defense genes GPX2, GPX3, TXN, NQO1 and HM0X1 in monolayer cultures of unsynchronized keratinocytes.

[0090] Furthermore, the Cananga odorata extract according to the invention possesses biological efficacy in protecting against oxidation and protein carbonylation in skin excipients subjected to environmental stress and photopollution. The demonstrated efficacies are as follows: - Decrease in the level of protein carbonylation in human skin expiants (whole skin) subjected to stress: urban particles and UVA irradiation. - Decrease in the level of protein carbonylation in the epidermal compartment of human skin expiants subjected to stress: urban particles and UVA irradiation. - Decrease in the level of protein carbonylation in the dermal compartment of human skin expiants subjected to stress: urban particles and UVA irradiation.

[0091] From this perspective, and according to another aspect, the invention relates to a cosmetic and / or dermatological composition comprising at least the extract of Cananga odorata according to the invention or obtained according to the process of the invention. The cosmetic and / or dermatological composition is suitable for topical application on the skin and / or mucous membranes, and / or hair and nails.

[0092] Preferably, in the composition of the invention, the extract according to the invention represents between 0.1% and 10%, preferably between 0.5% and 5% by weight of the composition.

[0093] The cosmetic and / or dermatological composition according to the invention can be presented in all the galenic forms normally used for a rinsed or non-rinsed topical application, for example in anhydrous form, in the form of an oil-in-water emulsion, a water-in-oil emulsion, a multiple emulsion, a silicone emulsion, a microemulsion, a nanoemulsion, a gel, an aqueous solution or a hydroalcoholic solution.

[0094] This composition may be more or less fluid and may be in the form of a white or colored cream, an ointment, a milk, a lotion, a serum, a gel, a cleansing base, or a stick. It may be used in numerous treatments for the skin, lips, and hair, including the scalp, in particular to protect and / or treat the skin, lips, and / or hair, and / or to apply makeup to the skin and / or lips.

[0095] The cosmetic and / or dermatological composition may contain excipients commonly used in the cosmetic and dermatological fields, such as fats, detergent and / or conditioning surfactants, emulsifiers and co-emulsifiers, hydrophilic or lipophilic gelling agents, preservatives, antioxidants, solvents, exfoliating agents, perfumes, fillers, hydrophilic and lipophilic sunscreens, colorants, acidic or basic neutralizing agents, pro-penetrating agents, and polymers. These types of excipients are all well known to those skilled in the art.

[0096] In practice, the quantities of these different excipients are those classically used in the fields considered, and the sum of the excipients can represent between 0.01% and 99.99% of the total weight of the composition.

[0097] Suitable fats include mineral oils, animal oils such as lanolin, vegetable oils, synthetic oils of natural and / or petrochemical origin such as isopropyl myristate, octyldodecanol, isostearyl isostearate, decyl oleate and isopropyl palmitate, and silicone oils such as cyclomethicone and dimethicone. Fatty alcohols, fatty acids, waxes and gums, and in particular silicone elastomers, may also be used as fats.

[0098] Suitable detergent and / or conditioning surfactants include non-ionic, anionic, cationic or amphoteric surfactants, and mixtures thereof, such as, for example, alkyl sulfates, alkyl ether sulfates such as sodium lauryl ether sulfate, alkyl betaines such as cocamidopropyl betaine, or quaternary ammonium salts.

[0099] Suitable emulsifiers and co-emulsifiers include, for example, polyglycerin and fatty acid esters, sucrose and fatty acid esters, sorbitan and fatty acid esters, oxyethylenated fatty acid and sorbitan esters, fatty alcohol and PEG ethers, glycerin and fatty acid esters, alkyl sulfates, alkyl ether sulfates, alkyl phosphates, alkyl polyglucosides, alkyl polypentosides, and dimethicone copolyols.

[0100] Suitable hydrophilic gelling agents may be cited for example carboxyvinyl polymers, acrylic copolymers (carbomers) such as acrylate / alkylacrylate copolymers, polyacrylamides, polysaccharides such as xanthan gum, guar gum, natural gums such as cellulose gum and derivatives, starches and their derivatives, clays and 2-acrylamido-2-methylpropane copolymers.

[0101] Suitable lipophilic gelling agents include, for example, modified clays such as bentones, metallic salts of fatty acids, hydrophobic silica, ethylcellulose, dextrose esters, hydrogenated castor oil and its derivatives, polyurethanes and magnesium salts.

[0102] Suitable preservatives may be cited for example benzoic, sorbic, propionic, salicylic, dehydroacetic acids and their salts, benzyl alcohol, ethylhexylglycerin, parabens, their salts and esters, triclosan, imidazolidinyl urea, phenoxyethanol, DMDM ​​hydantoin, diazolidinyl urea, chlorphenesin.

[0103] Suitable antioxidants may be cited for example chelating agents such as EDTA and its salts, sodium metabisulfite, salicylic, ascorbic and citric acids and their salts, sodium tartrate, sodium gluconate, carotenoids and tocopherols.

[0104] Examples of solvents that can be used in cosmetic composition (distinct from the extraction solvent) include water, ethanol, glycerin, propylene glycol, propanediol, butylene glycol, and sorbitol.

[0105] Suitable exfoliating agents include, for example, chemical exfoliants such as AHAs, and physical exfoliants such as natural or synthetic powders.

[0106] Suitable fillers may be cited for example talc, kaolin, mica, serecite, magnesium carbonate, aluminium silicate, magnesium silicate, organic powders such as nylon.

[0107] Suitable colorants may be cited for example lipophilic colorants, hydrophilic colorants, pigments and mother-of-pearls commonly used in cosmetic or dermatological compositions, and mixtures thereof.

[0108] Suitable neutralizing agents include basic agents such as, for example, sodium hydroxide, triethanolamine, aminomethyl propanol and potassium hydroxide, and acidic agents such as, for example, citric acid and lactic acid.

[0109] Suitable pro-penetrating agents include, for example, alcohols and glycols (ethanol, propylene glycol), ethoxydiglycol, alcohols and fatty acids (oleic acid), fatty acid esters, dimethyl isosorbide.

[0110] The composition of the invention may also contain, in addition to the extract according to the invention, other active ingredients. Suitable active ingredients include, for example, free radical scavengers or, more generally, antioxidants, whitening agents, pigments, emollients, moisturizers, anti-seborrheic agents, anti-inflammatories, anti-acne agents, keratolytic and / or desquamating agents, anti-wrinkle and tightening agents, draining agents, anti-irritant agents, soothing agents, vitamins and their mixtures, mattifying agents, anti-aging actives such as retinol, healing agents, antiseptics, and essential oils.

[0111] The process according to the invention, and the extract according to the invention thus make it possible to provide an effective active ingredient to stimulate the nocturnal and diurnal mechanisms of skin protection against aggressions and skin aging, while reducing or eliminating the allergenic risks associated with the use of the active ingredient, and offering a more eco-responsible solution.

[0112] The eco-design advantages of the invention are as follows: - The valorization of waste to extract an active ingredient of interest, exhibiting biological efficacy for skin care; - The use of agro-sourced solvents; - The non-toxicity of the extraction solvents for humans and the environment; - The use of solvents called "ingredient solvents", allowing the extract to be used directly in cosmetic and / or dermatological compositions; - Reducing the number of process steps with the possibility of eliminating energy-intensive solvent evaporation steps; - The possibility of avoiding the addition of additives or preservatives to stabilize the preservation of the extract; - The biodegradability of extraction solvents.

[0113] The invention and the advantages arising therefrom will become clear from the following embodiments. Examples

[0114] Example 1: Preparation and characteristics of NaDES solvents

[0115] The NaDES solvents Betaine / Glycerin / Propanediol in molar proportions of 1:1:5 and Sorbitol / Propanediol / Glycerin in molar proportions of 1:1:5 are obtained by mixing all the compounds for 30 minutes to 1 hour in a stirred reactor equipped with a double jacket through which a heat transfer fluid heated to 70 °C circulates. The solvents obtained are clear, colorless liquids. The propanediol used is propane-1,3-diol, and the betaine used is anhydrous.

[0116] These two solvents belong to the category of NaLTTM (Natural Low Transition Temperature Mixture) and more specifically to the category of NaDES (Natural Deep Eutectic Solvent). The methodology applied to characterize them is that described by Caprin B., Charton V., Rodier J.-D., et al. “Scrutiny of the supramolecular structure of bio-sourced fructose / glycerol / water ternary mixtures Towards green low transition temperature mixtures. Journal of Molecular Liquids, 2021, 337, pp. 116428.”

[0117] NaDES Betaine / Glycerin / Propanediol with a molar ratio of 1:1:5 is characterized by a single, low glass transition temperature: Tg = -108°C. NaDES Sorbitol / Propanediol / Glycerin with a molar ratio of 1:1:5 is characterized by a single, low glass transition temperature: Tg = -74°C. This glass transition temperature is determined by DSC analysis using a DSC-Q20 instrument (TA Instruments) equipped with a liquid nitrogen cooling system. The samples are placed in airtight aluminum capsules under a nitrogen flow (50 mL / min) and subjected to three temperature cycles: 1) from -140°C to 70°C at 10°C / min; 2) from 70°C to -140°C at 10°C / min; 3) from -140°C to 70°C at 10°C / min. The glass transition temperature (Tg) is determined from the second heating ramp by considering the midpoint of the thermal transition.

[0118] NaDES Betaine / Glycerin / Propanediol with a molar ratio of 1:1:5 (designated solvent BGP115) and NaDES Sorbitol / Propanediol / Glycerin with a molar ratio of 1:1:5 (designated solvent SPG115) are characterized by the existence of intermolecular hydrogen interactions, determined by 2D NOESY NMR analysis with an AVANCE III 400 MHz spectrometer in 1H. The intermolecular hydrogen interactions are evidenced by the presence of off-diagonal correlation spots. The supramolecular nature of the Betaine / Glycerin / Propanediol solvent in 1:1:5 molar ratios and of the Sorbitol / Propanediol / Glycerin solvent in 1:1:5 molar ratios is demonstrated.

[0119] Example 2: Preparation of extracts from Cananga odorata flower press cake

[0120] The plant-based raw material used for the preparation of the extracts is a Press cake obtained from the hydrodistillation of Ylang-Ylang flowers (Cananga odorata). corresponds to the solid residue of flowers taken at the end of the production of the complete essential oil, i.e. after approximately 14 to 18 hours of hydrodistillation.

[0121] This hydrodistillation cake is not currently used for any other purpose and therefore constitutes a final waste product for the essential oil manufacturing process. Its use as a plant-based raw material allows it to be reclassified from waste to co-product. As such, the impacts associated with the disposal of this waste, particularly environmental impacts, are avoided by reclassifying it as a co-product.

[0122] The fresh flowers used to prepare the flower press cakes are organically grown and were harvested in Mayotte in 2021. These flowers were processed by hydrodistillation on the day of harvest to extract the essential oil. The flower press cakes were then collected and dried. Finally, the dried flower press cakes were ground using a knife mill.

[0123] Process 1: Propanediol - IA Extract (PDO Solvent)

[0124] Dried and crushed flower cakes are extracted with 1,3-propanediol (denoted Propanediol) at a plant mass ratio of 5%, under continuous mechanical stirring, at a temperature of 80°C for 4 hours. A solid / liquid separation using nylon cloth is then carried out, and the resulting extract is clarified under pressure on cellulose plates and a cellulose acetate membrane to a cutoff of 0.2 µm. An extract of Cananga odorata is thus obtained and designated "Extract IA".

[0125] Process 2: NaDES BGP115 - Extract 2A (Solvent BGP115)

[0126] Dried and ground flower press cakes are extracted with the NaDES BGP115 solvent obtained in Example 1, with a plant mass ratio of 5%, under continuous mechanical stirring, and at a temperature of 80°C for a period of 4 hours. A solid / liquid separation on nylon cloth is then carried out, and the resulting extract is clarified under pressure on cellulose plates and on a cellulose acetate membrane to a cutoff point of 0.2 µm. An extract of Cananga odorata is thus obtained and designated "Extract 2A".

[0127] Process 3: NaDES SPG115 - Extract 3A (Solvent SPG115)

[0128] Dried and ground flower press cakes are extracted with the NaDES SPG115 solvent obtained in Example 1, with a plant mass ratio of 3%, under continuous mechanical stirring, and at a temperature of 80°C for a period of 4 hours. A solid / liquid separation on nylon cloth is then carried out, and the resulting extract is clarified under pressure on cellulose plates and on a cellulose acetate membrane to a cutoff point of 0.2 µm. An extract of Cananga odorata is thus obtained and designated "Extract 3A".

[0129] Example 3: Physico-chemical characteristics of extracts from flower cakes

[0130] The visual appearance, odor, color parameters L*a*b* and Gardner Index, and pH of extracts IA (Solvent Pdo), 2A (Solvent BGP115) and 3A (Solvent SPG115) from the flower cakes obtained in Example 2 are determined according to the following methods.

[0131] The visual appearance is determined for the pure extract placed in a colorless and transparent glass tube with a diameter of 1cm.

[0132] The odor is evaluated by an internal panel trained in olfactory evaluation methods.

[0133] The Gardner and L*a*b* color indices are measured on the pure vat extract 10mm square PMMA plastic optical path, with 10° observer and D65 illuminant (Konica Minolta equipment).

[0134] The pH is measured on the product diluted to Uz (m / m) in water, with an "Inlab Science" electrode (Metler equipment).

[0135] The complete results are shown in Table 1.

[0136] [Tables 1] Visual Appearance Odor Gardner Index Parameters Lsa'b* pH Extract 1A Clear orange / amber liquid Floral odor but not characteristic of Ylang-Ylang, with a "toasted" note. 8.5 83'5 / 64 4.8 Extract 2A Clear amber liquid Ylang-Yang odor slightly greener and less floral than that of the essential oil 10.6 69 / 16 / 74 5.4 Extract 3A Clear yellow-orange liquid Ylang-Yang odor, close to that of the essential oil with a "toasty" note 3.8 931-2 / 21 5.3

[0137] Extracts IA (Solvent Pdo), 2A (Solvent BGP115) and 3A (Solvent SPG115), having been prepared with similar extraction parameters but different solvents, exhibit different color and odor characteristics.

[0138] A number of compounds in Ylang-Ylang (Cananga odorata) essential oil, listed as allergens according to Regulation (EU) 2023 / 1545, having been identified in the prior art, the 57 molecules classified as allergens are also sought and quantified where applicable in each of these extracts. The analysis was performed by GC-MS. The limit of quantification for all the compounds analyzed is 1 mg / kg.

[0139] The results are shown in the following table 2.

[0140] [Tables2] Molecules Extract 1A Extract 2A Extract 3A mg / kg Benzaldehyde < 1 1 < 1 Benzyl alcohol 10 36 5 Benzyl benzoate 314 322 55 Benzyl carboxylate 7 6 <1 Benzyl salicylate 345 291 49 Beta-caryophyllene 14 2 <1 Cinnamyl alcohol 11 12 3 Isoeuphenol 7 7 1 Geranium acetate 11 8 <T TOTAL 719 883 113

[0141] Only 9 out of 57 compounds were detected in extracts IA (Pdo Solvent), 2A (BGP115 Solvent), and 3A (SPG115 Solvent) using the method employed. The remaining compounds on the list of 57 allergenic molecules were not detected. Extracts IA (Pdo Solvent), 2A (BGP115 Solvent), and 3A (SPG115 Solvent) contain less than 0.1% by weight of allergenic compounds and are considered to be non-allergenic.

[0142] Among these 9 compounds, two molecules are predominant in extracts IA (Pdo Solvent), 2A (BGP115 Solvent), and 3A (SPG115 Solvent): benzyl benzoate and benzyl salicylate. Extracts IA (Pdo Solvent) and 2A (BGP115 Solvent) have similar levels of each of the quantified molecules. Extract 2A (BGP115 Solvent) contains slightly fewer allergenic molecules than extract IA (Pdo Solvent). Extract 3A (SPG115 Solvent) has the lowest levels of these two molecules.

[0143] The total concentration of allergenic molecules in these extracts (< 0.1% by weight) is much lower than the levels analyzed in the essential oil, as well as in the absolute described in patent FR3110416. Indeed, manufacturers of Cananga odorata flower essential oil report total allergenic compound levels ranging from 10 to 46% by weight of essential oil. Extracts IA (PDO solvent), 2A (BGP115 solvent), and 3A (SPG115 solvent) therefore contain allergenic compound levels approximately 140 to 600 times lower than those of the essential oil, depending on the extract. Patent FR3110416 describes a composition in compounds classified as allergens of between 36 and 67% extract, i.e. concentrations much higher than that of extracts IA (Solvent Pdo), 2A (Solvent BGP115) and 3A (Solvent SPG115).These extracts IA (Pdo Solvent), 2A (BGP115 Solvent) and 3A (SPG115 Solvent) therefore present much lower allergenic risks than those that the essential oil might present.

[0144] Example 4: Effect of extract IA (solvent Pdo), extract 2A (solvent BGP115) and extract 3A (solvent SPG115) of Cananga odorata flower cakes on the transcriptomic expression of genes encoding antioxidant enzymes in normal human keratinocyte cultures.

[0145] Principle of method.

[0146] We used the qRT-PCR technique to measure the expression of the genes of interest GPX2, GPX3, TXN, NQO1 and HMOX-1 which respectively encode Glutathione Peroxidase 2, Glutathione Peroxidase 3, Thioredoxine, NAD(P)H Quinone Dehydrogenase 1 and Heme oxygenase-1, in monolayer cultures of normal human keratinocytes treated or not with extract IA (solvent Pdo) or extract 2A (solvent BGP115) or extract 3A (solvent SPG115) of Cananga odorata flower cakes.

[0147] Protocol.

[0148] Normal primary human keratinocytes are treated for 24 h with extract IA (Pdo solvent) or extract 2A (BGP115 solvent) or extract 3A (SPG115 solvent) of Cananga odorata at final concentrations of 0.5% and 0.25%. Untreated (NT) cells are used as a control. The cells are washed with phosphate-buffered saline and then lysed for RNA extraction. After RNA quantification and quality validation, the relative expression of the GPX2, GPX3, TXN, NQO1, and HMOX-1 genes is assessed by RT-qPCR.

[0149] The study is based on 3 independent experiments (n=3) for extracts IA (Pdo solvent) and 2A (BGP115 solvent) and on 2 independent experiments (n=2) for extract 3A (SPG115 solvent). The statistical test is a One-Way ANOVA followed by a Tukey post-test to compare the expression of antioxidant genes in untreated normal human keratinocyte cultures with respect to normal human keratinocyte cultures treated with extract IA (Pdo solvent) or extract 2A (BGP115 solvent) or extract 3A (SPG115 solvent) of Cananga odorata flower meal.

[0150] Results.

[0151] We quantified the transcriptomic expression of the GPX2, GPX3, TXN, NQO1, and HMOX-1 genes in monolayer cultures of normal human primary keratinocytes treated or untreated with extract IA (Pdo solvent), extract 2A (BGP115 solvent), or extract 3A (SPG115 solvent). Results are expressed as a percentage relative to NT cells. Values ​​are expressed as mean ± standard deviation over the 2 or 3 experiments (n=2 or n=3).

[0152] The results for extract IA (solvent Pdo) are recorded in the following tables 3 to 7.

[0153] [Tables3] GPX2 Extract IA (Pdo) Exp 1 Exp 2 txp 3 Untreated Solvent Pdo 019587 IW 0.5% Of25% Untreated Solvent Pdo 019587 (PDpJ 0.5% 0.25% fton treated Solvent py.o 015048 ÊM 0.5% 104 103 94 1-66 132 173 543 415 550 448 415 183 101 151 153 171 323 392 379 366 444 103 ICI 95 164 163 478' 491 335 Average 1QO 175 547 426 160 382 379 100 166 451 Standard deviation 6 s 5 19 4 10 61 18 4 5 58 % stimulation at 0.5% vs NT1SQ% n=3 experiments. 449% ± 82 {"** p<0.001") Statistical test vs untreated: One-WayANGAA followed by a post-test of "" p <D,<3apl

[0154]

[0155] Table 3: GPX2 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cakes at 0.5% and 0.25%. [Tables 4] Extract IA (Pdo) Exp 1 Exp 2 Exp S Won trotted Solvent Pdo 019587 {PDoJ 0.5% 0.25% Untreated Solvent iPdo D19S87 (PDo) 0.5% 0.25% treated Pdo DÎKM8 (PDo) 0.5% 101 97 law 144 134 125 465 213 422 194 133 105 100 95 162 148 148 272 185 385 174 422 92 93 131 128 126 436 457 344 Mean 100 134 444 198 100 153 360 ISO ioe 128 416 Standard deviation 2 10 30 13 5 8 78 8 14 3 64 % stimulation at 0.5% vs NT 100% 9=3 experiments 402%±66 r*** p0.001) Statistical test vs untreated: One-Way ANOVA followed by a Tokey post-test, p<0.0003

[0156] Table 4: GPX3 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cake at 0.5% and 0.25%.

[0157] [Tables5] Exp 1 Exp 2 Exp 3 < Extract IA (Pdo) Untreated Solvent Pdo D19587 (PDS) $.5® 0.25% Untreated Solvent Pdo 019587 (PDo| 0.5% 0.25% Untreated Solvent Pdo D19O48 (PDü) 0.5% 102 33 237 133 104 85 253 142 107 94 20ü .105 95 232- 136 100 37 2 SC 128 Wi 105 223 33 91 127 95 84 303 92 98 "173 TXN Average 100 93 235 132 100 85 282 135 100 BS 200 Standard deviation 7 2 4 5 5 2 26 10 8 6 2S % Q,S% stimulation vs 2«%±43 ("*• p<0.0001) nt ira% ! is statistical vs untreated: One-Way ANOVA followed by a Tukey post-test, experiments p<0.0001

[0158] Table 5: Expression of the TXN gene (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0159] [Tableauxô] Exo 2 Exp 4 Extract IA jPcfa) Untreated Soivarrt pdo D19587 (PDo) O,S% 0.25% Untreated Solvent Pria 019587 (PDo) 0.5% 0.25% Untreated Solvent Pdo D19048 (PDo] 0.5% 101' .133. 388 375 182 134 430 322 ICI 125 322 W7 155 416 .370 104 135 458 296 110 143 320 91 149 330 94 128 477 9G S135 275 NQO1 Average 100 145 432 375 132 4SS 309 100 135 30$ Standard deviation 8 12 20 5 5 4 24 18 30 9 27 % stimulation 8.5% vs 386%±7 (,,M pc0.000l} NT 100% n=3 experiments Statistical test vs untreated: One-Way ANOVA followed by post-Tukév test, **** p <O,®EU

[0160]

[0161] Table 6: NQO1 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cake at 0.5% and 0.25%. [Paintings?] HMOX -i Extract !To{Pdo) Exp 1 Exp 2 Exp 3 Untreated Solvent Pdo D195S7 (PDo) 0.5 0.25 % % Untreated Solvent Pdo 019587 (PDO) 0.5 8.25 % % Untreated Solvent Pdo D1904 8 {PDO) 0.5% 92 113 130 167 145 12Q 444 121 441 3S4 . 79 99 129 151 138 173 307 572 906 99 101 157 165 182 259 Mean» e 10Û 149 120 423 W2 154 573 493 100 169 196 Standard deviation 11 16 ï 34 25 18 305 112 1 13 89 % stimulat! on 386%±287 (ns) at 0.5% vs MT100% eXpérieriC Statistical test vs untreated; One-way ANOVA followed by a Tukey post-test, ns {not significant)

[0162] Table 7: Expression of the HMOX-1 gene (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cake at 0.5% and 0.25%.

[0163] The AI ​​extract (Pdo solvent) of Cananga odorata flower cake obtained according to the invention induces a significant increase in the transcriptomic expression of the GPX2, GPX3, TXN, and NQO1 genes at 0.5% in 2D cultures of normal human primary keratinocytes. The extraction solvent alone does not induce an increase in the expression of the GPX2, GPX3, TXN, and NQO1 genes (data not shown). The AI ​​extract (Pdo solvent) of Cananga odorata exhibits antioxidant capacity by activating several target genes encoding antioxidant enzymes.

[0164] The results for extract 2A (solvent BGP115) are recorded in the following tables 8 to 12.

[0165] [Tables8] E*p 1 Exs 7 Ew 3 Extract 2A. (BGP115) Untreated Scfvant BGP1Î5 D19S48 (BGP115J 0.5% 0.25% Untreated Soivarst BGP115 D19548 (BGPllSj 0.5% 0.25% Untreated Servant BGP115 D19S48 (8GP115) 0.5% 104 127 414 522 103 130 353 420 103 98. 430 103 124 426 480 101 131 353 4® 101 105 375 94 133 4S3 95 Ï3S 339 95 481 GPXZ Average 100 128 420 498 100 132 350 415 160 102 429 Standard deviation & 5 8 21 4 3 10 21 4 5 53 » stimulation at 0.5% vs 397% ± 49 (**** p <O,0OOl) NT 160% n=3 expériences Test statistique « Ton traité: C ne -Way ANOVA suivi d'un post-test de Tuitey, **“ p<Q.æcfi

[0166] Table 8: GPX2 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0167] [Tables9] Extract 2A {SGP115J Exp 1 Ex» 2 Exp 3 Untreated Solvent BGP115 313548 (BGP11S) 0.5% 0.25% Untreated Solvent BGP115 D19S48 (BGP115) 0.5% 0.25% Untreated Solvent BGP115 D19548 (BGP115) 0.5% 101 37 101 W5 94 336 210 316 204 138 IBS 100 95 120 108 105 290 133 324 139 290 10 U) L" wm £ 112 9G 451 344 587 GPX3 Mean 100 95 326 204 IGG 111 301 191 iœ 101 461 Standard deviation 2 10 14 6 5 8 20 3 16 122 % stimulation at 0.5% vs NT 100% 357% ± 102 {*«* pcO.DOOl) n=3 experiments Statistical test vs untreated; One-Way ANOVA followed by a Tukey post-test, ” '' p <O,0Wi

[0168]

[0169] Table 9: GPX3 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5% and 0.25%. [Tables 10] Exp 1 ;txp Exp 3 Extract 2A (SGP115) Untreated Solvent BGP115 D1954S (8GP115J 0.5 0.2% 5% Untreated Following BGP11 5 D19548 {8GP115j 0.25 0.5% % Untreated Solvent 8GP115 019548 (BGP11Sj 0.5% 1G2 9Q 205 260 104 94 249 150 107 S& 240 1Q6 34 200 150 100 99 277 150 101 S4 218 93 83 160 95 96 283: 160 92 235 Mean 100 86 203 1.57 100 96 270 157 100 91 248 Standard deviation 7 4 4 6 5 3 18 6 8 10 34 % stimulation n 245% ± 35 {***' p<0.9001) at 9.5% vs NT 100% n=3 Statistical test vs untreated: One-Way ANOVA followed by a Tukey post-test, “*** p<0.9001

[0170] Table 10: Expression of the TXN gene (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0171] [Tableauxll] NQOl Extract 2A (BGP115) Exp 1 Exp 2 Exp 3 Untreated Savant BGP11 5 D19548 (BGP115) 05 0.25% % Untreated Solvent 8GP11 5 D19548 (BGP115) 0.5 0.25% % Untreated Solvent SGP11 S 01954 S (BGP115) 0.5% 101 107 91 119 123 123 388 468 421 437 495 102 :W 94 12:4 144 126 487 390 477 382 458 101 12.0 90 103 86 431 437 499 Average 1OS 123 405 467 100 131 474 386 100 95 456 Standard deviation 8 5 23 29 5 11 15 6 10 12 38 » stimulation at 0.5¾ vs biT10ü% n=3. experiments-: 458% ± 37 P'””' p <O,OQOl) Test statistique vs non traité: One -Waÿ AMJVA Suivi d'un post-test de Tukey, p<0,0001

[0172] Table 11: NQO1 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cake at 0.5% and 0.25%.

[0173] [Tables 12] Exp 1 Exp 2 Extract 2A {8GP115} Untreated Sotvaot BGP115 D19548 (BGP11S) 0.5% 0.25% Untreated SoîvaRt BGP115 D19548 {BGP115} 0.5% 0.25% 96 92 113 115 118 400 535 520 499 285 450 79 99 ±29 133 121 119 351 445 36 372 754 HMOX- 1 Mean 100 117 402 495 102 126 404 409 Standard deviation 11 2 118 43 25 10 341 52 % stimulation at 0.5% vs NT 100% 403% ±228 p<0.01) n-2 experiments s is statistical vs untreated: One-way ANOVA followed by a Tukey post-test, p <Q,01

[0174] Table 12: Expression of the HMOX-1 gene (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0175] Extract 2A (solvent BGP115) of Cananga odorata flower cake obtained according to the invention induces a significant increase in the transcriptomic expression of the GPX2, GPX3, TXN, NQO1, and HMOX-1 genes at 0.5% in 2D cultures of normal human primary keratinocytes. The extraction solvent alone does not induce an increase in the expression of the GPX2, GPX3, TXN, NQO1, and HMOX-1 genes (data not shown). Extract 2A (solvent BGP115) of Cananga odorata exhibits antioxidant capacity by activating several target genes encoding antioxidant enzymes.

[0176] The results for extract 3A (Solvent SPG115) are recorded in the following tables 13 to 15.

[0177] [Tables 13] GPX2 Extract 3A {SPG115) Eœ 1 Ex a 2 ton treated Solvent SPG115 019395 (5PG115) 0.5% 0.25% My treated Solvent SPG115 019305 (SPG115) 0.5% 0.25% SS 97 128 93 101 222 148 219 159 230 153 37 9S 10? 130 97 110 178 118 148 203 159 Average ISO 10? 224 153 108 112 187 142 Standard deviation 4 18 6 S 6 17 23 21 % stimulation at 0.5% VS NT 100% n=2 209% ± 24 (** p<0.0001j Statistical test vs untreated: One-Way ANOVA followed by a post-test of Tufey, Σ<0.0301

[0178] Table 13: GPX2 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 3A (solvent SPG115) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0179] [Tables 14] Extract 3A(SPGH5) Exp 1 Ex? 2 Untreated Solvent SPG115 019305 {SPG115} 0.5% 0.25% untreated Solvent SPG115 M9305 (SPG115J 0.5% 0.25% 103- 96 101 109 97 100 251 158 225: 174: 230 154 95 95 111 111 104 110 202 128 216- 148 228 147' Mean 100 182 235 162 100 108 209 141 Standard deviation 4 6 14 11 9 4 7 % stimulation at 0.5% VS NT 100% 222% ± 18 J • " ' p<0.0001) Statistical test vs untreated: One-Way ANQVA followed by post-Tukeÿ test, n=2 " - p <o,æoi

[0180] Table 14: NQO1 gene expression (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 3A (solvent SPG115) of Cananga odorata flower cake at 0.5% and 0.25%.

[0181] [Tables 15] Ex □ 2 Exp 2 Extract 3 A (SPG115) treated Solvent SPG11S 019305 (SPG115J 09% 0.25% treated Solvent SPG115 019305 (SPG115J 0.5% 0.25% 93 126 144 95 101 145 131 93 109 97 156 120: 92 84 158 121 SS 121 172 140 108 126 172 136 HMOX-1 Mean ICO ns 157 128 100 118 154 117 Standard deviation 8 16 14 21 8 31 21 22 % stimulation at 0.5% vs m 100% 156%±16j* sp<0.0l ) n ? Statistical test vs untreated: One-Way ANOVA followed by a Tukey post-test.

[0182] Table 15: Expression of the HMOX-1 gene (in % relative to the untreated condition) in monolayer cultures of normal human primary keratinocytes treated for 24h with extract 3A (solvent SPG115) of Cananga odorata flower cakes at 0.5% and 0.25%.

[0183] Extract 3A (SPG115 solvent) of Cananga odorata flower cakes The extract obtained according to the invention induces a significant increase in the transcriptomic expression of the GPX2, NQO1, and HMOX-1 genes at 0.5% in 2D cultures of normal human primary keratinocytes. The extraction solvent alone does not induce an increase in the expression of the GPX2, NQO1, and HMOX-1 genes (data not shown). Extract 3A (solvent SPG115) of Cananga odorata exhibits antioxidant capacity by activating several target genes encoding antioxidant enzymes.

[0184] Example 5: Effect of extract IA (solvent Pdo) and extract 2A (solvent BGP115) of Cananga odorata flower cakes on the activation of the Nrf2 antioxidant pathway, and more specifically on the translocation of the Nrf2 transcription factor into the nucleus of unsynchronized normal human keratinocytes.

[0185] Principle of the method.

[0186] We used in situ immunostaining coupled with image analysis to measure the nuclear expression of the transcription factor Nrf2 in monolayer cultures of normal human keratinocytes treated or not with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes.

[0187] Protocol.

[0188] Normal human keratinocytes at 70-80% confluence are treated for 6 hours with extract IA (Pdo solvent) or extract 2A (BGP115 solvent) of Cananga odorata at final concentrations of 0.5%, 0.25%, and 0.1%. Untreated (NT) cells are used as controls. The normal human keratinocytes are then washed with phosphate-buffered saline before being fixed (Antigenfix), permeabilized with 0.1% Triton for 5 minutes, and saturated with 1% BSA (bovine serum albumin) for 1 hour. The cells are incubated with the primary antibody (anti-Nrf2) overnight at 4°C, washed with PBS buffer, and incubated with the Alexa 594 fluorochrome-bound secondary antibody for 1 hour. Fluorescence is read on a Cytation 5 imaging spectrophotometer (Biotek) with appropriate filters. Fluorescence measurements by image analysis are performed using common acquisition parameters.

[0189] The study is based on 3 independent experiments (n=3). The statistical test is a One-Way ANOVA followed by a Tukey post-test to compare the nuclear expression of Nrf2 in untreated normal human keratinocyte cultures versus normal human keratinocyte cultures treated with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes.

[0190] Results.

[0191] We quantified the nuclear expression of Nrf2 in the nuclei of normal human keratinocytes treated or untreated with extract IA (Pdo solvent) or extract 2A (BGP115 solvent) of Cananga odorata flower meal. Results are expressed as the percentage of Nrf2-translocated cells per total cell count. Values ​​are expressed as mean ± standard deviation across three independent experiments (n=3). A one-way ANOVA followed by a Tukey post-test was performed versus the untreated condition.

[0192] The results are recorded in the following table 16.

[0193] [Tableauxlô] Extract IA, Pdo: % reflufe Nrf2 trarôio^uées on the number of total ceikdes Extract IA (Prfo) Exp 1 Ex o 2 Ex P 3 Untreated Solvani Pdo 819587 {PDo| 0 / 2 0, 0.5% 5% 1% Untreated SoSvafit Pdo D19S87 {PDo} 0.2 O, 0.5% 5% 1% Untreated Solvent Pdo D19587(PDo} 0.2 0, 0.5% 5% 1% 6 7 19 13 16 43 36 34 56 42 34 46 43 33 12 .11 8 28 21 13 78 53 28 76 54 24 68 45 23 21 12 18 21 19 16 57 41 " 64 32 Average 8 18 48 40 34 10 21 74 52 25 17 18 58 33 Standard deviation 3 2 7 4 1 2 s 5 3 3 5 3 5 5. - % Ntf2 translocated cells at 0.5% on the total number of cells 60% ± 12p <O,OOGl) expériences Test statistique vs non traité; One -Way ANÛVA suivi d’un post-test de Tukey, *♦** p<0,£> 301

[0194] Table 16: Translocated Nrf2 cells (as a % of total cells) in monolayer cultures of normal human keratinocytes treated for 6 h with IA extract (Pdo solvent) of Cananga odorata flower meal at 0.5%, 0.25%, and 0.1%. One-way ANOVA followed by a Tukey post-test was performed versus the untreated condition.

[0195] The IA extract (Pdo solvent) of Cananga odorata flower cake obtained according to the invention significantly increases the translocation of the transcription factor Nrf2 (60% of cells translocated to 0.5% vs. 12% for the untreated condition) into the nucleus of normal human keratinocytes compared to the untreated condition. The extraction solvent alone does not induce an increase in Nrf2 translocation into the nucleus of keratinocytes (data not shown). The IA extract (Pdo solvent) activates the Nrf2 antioxidant pathway in the nucleus of unsynchronized keratinocytes.

[0196] The same experiment is carried out with extract 2A (solvent BGP115). The results are recorded in Table 17 below.

[0197] [Tables 17] Extract 2A, 8GPH5;“Sr.eiluSes Nrf2 tisBsiaqiiéeapar ratio of total cells Extract 2A (66P115J Exp_l Exp_2 Exp_3 Untreated Soivant BGP11 S »19548 (BGP115) 0.5 0.25 0.1 % % % treated Solvent BGP11 5 D19S4SJBGP11S} O.2S 0.1 0.5% % » Untreated Next BGP11 S 019548 (SGP11S) 0.25 0.1 0.5% % % II S 16, 15 14 49 43 39 45 4S 40 4? 15 12 50 .53 33 40 45 26 40 46 26 12 19 18 11 7 78 53 59 47 §9 42 Mean 8 15 47 42 39 10 16 43 43 28 17 12 72 47 Standard deviation 3 1 2 7 1 2 5 6 4 4 5 6 5 6 % of Nif2 cells translocated to 0.5% on the total number of cells 54% ± 14 (*'** n=3 experiments Statistical test vs. untreated: One-Way AWORK followed by a Tukey post-test, **** p<0.0001; ***

[0198] Table 17: Translocated Nrf2 cells (as a % of total cells) in monolayer cultures of normal human keratinocytes treated for 6 h with extract 2A (solvent BGP115) of Cananga odorata flower meal at 0.5%, 0.25%, and 0.1%. One-way ANOVA followed by a Tukey post-test was performed versus the untreated condition.

[0199] Extract 2A (solvent BGP115) of Cananga odorata flower cake obtained according to the invention significantly increases the translocation of the transcription factor Nrf2 (54% of cells translocated to 0.5% vs. 12% for the untreated condition) into the nucleus of normal human keratinocytes compared to the untreated condition. The extraction solvent alone does not induce an increase in Nrf2 translocation into the nucleus of keratinocytes (data not shown). Extract 2A (solvent BGP115) activates the Nrf2 antioxidant pathway in the nucleus of unsynchronized keratinocytes.

[0200] Extract IA (solvent Pdo) and extract 2A (solvent BGP115) of Cananga odorata flower cakes obtained according to the invention have almost equivalent efficiencies on the translocation of the Nrf2 transcription factor into the nucleus of unsynchronized keratinocytes.

[0201] Example 6: Effect of extract IA (solvent Pdo) and extract 2A (solvent BGP115) of Cananga odorata flower cakes on the activation of the Nrf2 antioxidant pathway, and more specifically on the translocation of the Nrf2 transcription factor into the nucleus of synchronized normal human keratinocytes, in “night” mode.

[0202] 6.1 Study of the synchronization of normal human keratinocytes.

[0203] A monolayer model of keratinocytes in "night" mode was previously developed. For this purpose, keratinocyte cultures were synchronized (treatment with dexamethasone at IpM for 2 h) to model the circadian rhythm, i.e., the biphasic expression of the "night" (BMALI) and "day" (PER3) clock genes. Monitoring the expression of the clock genes from T0h to T72h, with a pause every 6 h using RT-qPCR (Polymerase Chain Reaction), allowed us to demonstrate that at T30h and T54h, the expression of the "night" gene (BMALI) was antiphase with the expression of the "day" gene (PER3). It should be noted that the amplitude between "night" and "day" expression was greater at T30h.

[0204] For the in vitro efficacy test, monitoring the translocation of the transcription factor Nrf2 in the nucleus of keratinocytes in "night" mode, we decided to perform the treatments with extracts IA (Pdo solvent) or 2A (BGP115 solvent) of Cananga odorata flower cakes between T24h and T30h (i.e., 6 hours of treatment equivalent to the initial protocol), that is, around the peak expression of the "night" clock gene BMALL

[0205] 6.2 Efficacy of extracts IA (solvent Pdo) or 2A (solvent BGP115) on translocation of Nrf2 into synchronized normal human keratinocyte cultures, in “night” mode.

[0206] Principle of the method.

[0207] We used the qRT-PCR (Polymerase Chain Reaction) technique to measure the expression of the BMAL1 and PER3 genes in monolayer cultures of normal human keratinocytes synchronized and treated or not with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes.

[0208] We used in situ immunostaining coupled with image analysis to measure the nuclear expression of the transcription factor Nrf2 in monolayer cultures of normal human keratinocytes synchronized and treated or not with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes.

[0209] Protocol.

[0210] Normal human keratinocytes with 70-80% confluence are synchronized for 2h with dexamethasone at IpM. The end of dexamethasone treatment corresponds to time TOh.

[0211] For the study of cell synchronization, and more specifically for monitoring the antiphase expression of the "day" and "night" genes (PER3 vs. BMAL1), kinetics are performed at T6h, T24h, and T30h. At each kinetic time point, the cell mats are rinsed with a phosphate buffer and then lysed with a highly denaturing buffer before purification by complementary hybridization of the total RNAs with The RNA extraction kit "MagMax™-96 Total RNA Isolation Kit" (Invitrogen). After quantification and validation of RNA quality, the relative expression of the "day" (PER3) and "night" (BMALI) genes is evaluated by RT-qPCR.

[0212] qRT-PCR is the most sensitive method for detecting and quantifying messenger RNA (mRNA) at the cellular level. RNA is first reverse-transcribed (RT) into complementary DNA (cDNA), which is more stable than RNA. It is the cDNA that is used to perform PCR. PCR allows real-time monitoring of the PCR amplification process by detecting the fluorescence emitted by the newly formed PCR products. Detection was performed using SYBR Green, a fluorescent organic compound that binds to nucleic acids. The relative quantification of the newly formed transcripts is performed using the tCA calculation method by normalizing the obtained expressions against a housekeeping gene, thus allowing comparison between two samples.

[0213] To monitor the translocation of the transcription factor Nrf2 into the nucleus of synchronized keratinocytes, cells are treated for 6 h (between T24h and T30h, at the peak expression of the "night" gene BMAL1) with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata at a final concentration of 0.5%. Untreated (NT) cells are used as a control. Normal human keratinocytes are then washed with phosphate-buffered saline before being fixed (Antigenfix), permeabilized with 0.1% Triton for 5 minutes, and saturated with 1% BSA (bovine serum albumin) for 1 hour. The cells are incubated with the primary antibody (anti-Nrf2) overnight at 4°C, washed in PBS buffer, and incubated with the secondary antibody bound to the Alexa 594 fluorochrome for 1 hour. Fluorescence is read on a Cytation 5 imaging spectrophotometer (Biotek) with appropriate filters.Fluorescence measurements by image analysis are performed using common acquisition parameters.

[0214] The study is based on 3 independent experiments (n=3). The statistical test is a One-Way ANOVA followed by a Tukey post-test to compare the nuclear expression of Nrf2 in untreated normal human keratinocyte cultures versus normal human keratinocyte cultures treated with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes.

[0215] Results.

[0216] We have quantified the transcriptomic expression of the "night" gene (BMAL1) and the "day" gene (PER3) in normal human keratinocytes, previously synchronized with dexamethasone.

[0217] Antiphase expression of the "night" gene (BMAL1) and the "day" gene (PER3) is confirmed in the T24h to T30h time window. Therefore, keratinocytes are synchronized and in "night" mode during this time window, which corresponds to 6 hours of treatment of keratinocytes with extracts IA (solvent Pdo) or 2A (solvent BGP115) of Cananga odorata flower cakes.

[0218] After validating keratinocyte synchronization in "night" mode, we quantified the nuclear expression of Nrf2 in the nuclei of normal human keratinocytes treated or untreated with extract IA (Pdo solvent) or extract 2A (BGP115 solvent) of Cananga odorata flower meal. The results are expressed as the percentage of Nrf2-translocated cells per total cell count. Values ​​are expressed as mean ± standard deviation over 3 independent experiments (n=3). A one-way ANOVA followed by a Tukey post-test was performed versus the untreated condition. The results are shown in Table 18 below.

[0219] [Tables 18] Extract IA, Pdo % cells Nrf 2 translocated on the total number of cells Extract IA (Pdo) Exp 1 Exp 2 Exd 3 Untreated Servant Pdo D19587 (PDo) 0.5% Untreated Solvent Pdo D19587 (PDo) 0.5% Untreated Solvent Pdo «3.5% 13.9 6.3 6.7 11.8 8.2 3.2 a 14.4' 56.6. 36.5 51.7 43.6 35.5 40.1 334 31.2 40.7 33.5 33.9 39 53.4 53.2 51.5 624 26.9 23.5 24.1 . 173 23.9 28.3 434 28.4 454 43 504 51.4 Mean w 12 59 35 33 58 24 27 49 Standard deviation 4 ï ■9 4 3 6 5 3 2 "NrfZ cells translocated at 0.5% of the total number of cells 52 S ± 5 (* p<0.001) in=3 experiments Statistical test vs untreated: One-Way ANOVA followed by a Tukey post-test, 4 ** p<3.001

[0220] Table 18: Translocated Nrf2 cells (as a % of total cells) in overnight cultures of normal human keratinocytes in monolayer, treated for 6 h (between T24h and T30h) with 0.5% IA extract (Pdo solvent) of Cananga odorata flower meal. One-way ANOVA followed by a Tukey post-test was performed vs. the untreated condition.

[0221] The IA extract (Pdo solvent) of Cananga odorata flower cake obtained according to the invention significantly increases the translocation of the transcription factor Nrf2 (52% IA extract at 0.5% vs. 23% for the untreated condition) into the nucleus of normal human keratinocytes, previously synchronized with dexamethasone, compared to the untreated condition. The extraction solvent alone does not induce an increase in Nrf2 translocation into the keratinocyte nucleus (data not shown). The IA extract (Pdo solvent) activates the Nrf2 antioxidant pathway in the The nuclei of synchronized keratinocytes are in "night" mode. The results are recorded in Table 19 below.

[0222] [Tables 19] Extract 2A, SG.P11S: % of transformed Wrf2 compared to the number of transformed cells Extract 2A ■ 8GP115) Exp 1 Exp 2 Exp 3 Untreated Solvent 8GP11 5 D19548 (BGP11S} 0.5% treated Solvent BGP11 D19548 {BGPHS| $5% Untreated Solvent 8GP11 5 D19548 {BGP115J 0.5% 13.9 63 6.7 1^3 8.7 12 16.2 43.6 25.5 23.6 36.5 40.1 35.2 31.2 42 323 353 623 523 56.6 503 263 23.5 243 173 323 33>6 363 313 54 493 43.6 38.4 fsteyeime 10 il 35 35 35 56 24 29 46 Standard deviation 4 4 10 4 S 5 5 4 7 SS celiirfes N?12 transioquëes at 0.5% on % number of total tests n=3 experiments 46% ±10 (*p<0.05J Statistical test vs untreated: One-Way ÀKC^Âsuivi d'an posttesi deTukey, p<0.05

[0223] Table 19: Translocated Nrf2 cells (as a % of total cells) in monolayer cultures of normal human keratinocytes in "night" mode treated for 6 h (between T24h and T30h) with extract 2A (solvent BGP115) of 0.5% Cananga odorata flower meal. Values ​​are expressed as mean ± standard deviation over 3 independent experiments (n=3). One-way ANOVA followed by a Tukey post-test was performed versus the untreated condition.

[0224] Extract 2A (solvent BGP115) of Cananga odorata flower cake obtained according to the invention significantly increases the translocation of the transcription factor Nrf2 (46% in extract 2A at 0.5% vs. 23% for the untreated condition) into the nucleus of normal human keratinocytes, previously synchronized with dexamethasone, compared to the untreated condition. The extraction solvent alone does not induce an increase in Nrf2 translocation into the nucleus of keratinocytes (data not shown). Extract 2A (solvent BGP115) activates the Nrf2 antioxidant pathway in the nucleus of synchronized keratinocytes, in "night mode".

[0225] Extract IA (solvent Pdo) and extract 2A (solvent BGP115) of Cananga odorata flower cakes obtained according to the invention retain their level of effectiveness on the translocation of the transcription factor Nrf2 into the nucleus of synchronized keratinocytes, in "night" mode. Thus, extracts IA (solvent Pdo) and 2A (solvent BGP115) function in both "day" and "night" modes.

[0226] Example 7: Effect of extract IA (solvent Pdo) and extract 2A (solvent BGP115) of Cananga odorata flower cakes on the level of oxidation, carbonylation of proteins in human skin expiants stressed by photopollution (exposure to urban particles followed by UVA irradiation) (Service provided by Oxiproteomics).

[0227] Principle of method.

[0228] Oxiproteomics used the patented "OxiProteomics® fluorescent probe" technique to visualize in situ oxidatively damaged (carbonylated) proteins on skin expiants treated with extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5%, 1% and 2%, before being exposed to pollution and UVA.

[0229] Protocol.

[0230] Skin expiants were obtained with the informed consent of a 30-year-old male (Caucasian, phototype 11-111) who had undergone abdominal surgery. Extract IA (solvent Pdo) or extract 2A (solvent BGP115) of Cananga odorata flower cake was applied topically to the skin expiants (30 µl / cm²) at concentrations of 0.5%, 1%, and 2% for 24 hours. The skin expiants were then subjected to stress conditions: a solution containing fine dust (0.375 µg / cm²; Fine Dust PM 10-like; ERMCZ100 Sigma-Aldrich) was applied topically for 30 minutes before UVA irradiation (6 J / cm²) using the OxiProteomics® irradiation system.

[0231] After exposure to stress, the skin expiants are transferred to fresh culture medium for 2 hours. Untreated, unstressed skin expiants are used as a control. After 2 hours of recovery, the skin samples are transferred to a cryogenic embedding medium, OCT (Optimal Cutting Temperature compound) for cryopreservation, frozen in liquid nitrogen, and stored at -80°C until analysis.

[0232] 5 µm skin sections are obtained using a cryostat and then fixed. Oxidation-damaged (carbonylated) proteins are labeled using a functionalized fluorescent probe, the "OxiProteomics® fluorescent probe" (a technique patented by Oxiproteomics), to bind specifically to carbonyl groups and DAPI for nuclear labeling. Fluorescence images are collected with an epifluorescence microscope (EVOS M5000 Imaging System) and analyzed with ImageJ software (Schneider, 2012).

[0233] The intensity of carbonylation was obtained by integrating the specific fluorescence signal normalized by the evaluated area. In each image, the carbonylation level was quantified independently for the different anatomical compartments (dermis, epidermis) and for the whole tissue. Three images per condition (one image per expiry) were used to quantify the levels of carbonylation. The mean value and standard deviation are obtained per condition and per anatomical compartment. Data processing and statistical analysis are performed using graphPad Prism.

[0234] Results.

[0235] In situ detection of protein oxidation and carbonylation levels was performed by epifluorescence microscopy. Densitometric analysis of the in situ carbonylation signal was performed using ImageJ software. Three images per condition were analyzed. The results are presented for the whole skin and by anatomical compartment, namely the epidermis and the dermis. Stress, the application of urban particles to the surface of skin excipients followed by UVA irradiation, induces an increase in the level of protein carbonylation in the tissues.

[0236] The results for extract IA (solvent Pdo) are recorded in the following tables 20 to 22.

[0237] [Tables20] Extract IA (following Pdo} Epidermis + Dermis Protein oxidation level % (normalized against the stressed condition) My treated Stress (PM + UV-A) 019587 (Pdpj (2%) + Stress 019587 (PdoHl%) + Stress D19587(Pdo} (0.5%) + Stress 43.5 91.7 61.3 54;O 54.3 4&2 97.5 38.4 64.5 6 2.3 n=3 55.5 110.8 52.5 30.4 58.8 Mean 51 100 51 66 59 EC 4 10 12 13 4 % decrease protein carbonylation level to 6.5% vs Stress (PM + UV-A)} h=l experiment is statistical vs stress: AMOVA &. QpmétVs muttipie comparisons test

[0238] Table 20: Quantification of the level of oxidation, protein carbonylation (%) in the epidermis and dermis of skin explants treated for 24h with IA extract (Pdo solvent) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with topical application of urban particles (0.375qg / cm2; Fine Dust PMIO-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0239] [Tables21] Extract IA jsoivant Pdo) Epidermis Protein oxidation level % (normalized relative to the stressed condition) Untreated Stress (PM + UV-A) 019587 {Pdo} (2%) + Stress D19587 (Pdo) ("£) + Stress D19587 {Pdo} {0.5%) + Stress n=ln=2 0=3 51 / 3 52.9 56.2 104..0 93.8 1G2.2 63.1 48.8 54.9 53.7 63.3 67..3 58.3 65.9 54.8- Mean E€ 53 3 100 S 56 7 61 9 60 6 % decrease in protein carbonylation rate at Q,S% vs Stress {PM + UV-A}) nJ experiment p0.OOl) Statistical test vs stress: ANOVA & Dunnett's multiple comparisons test

[0240] Table 21: Quantification of the level of oxidation, carbonylation of proteins (%) in the epidermis of skin explants treated for 24h with the IA extract (Pdo solvent) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with the topical application of urban particles (0.375qg / cm2; Fine Dust PM10-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0241] [Tables22] Dermis Protein oxidation level % (untreated compared to stressed content) Untreated Stress (PM + UV-A) D19S87 (Pdo; 2%) s-Stress D19S87 (Pdo) (1%) + Stress D19S87 (Pdo) ¢0.5%) + Stress n=1 ■ 4G,7 100 / 2 5$?1 44.9 49.0 fi=2 43.1 S3Z3 35.6 53.5 55.0' n=3 49.8 106.5 41.9 78.7 60.0 Mean 45 100 43 59 55 EC 5 7 7 18 5 % direction level of protein carbonylation s O,S% vs Stress (PM + UV-A) n=î experiment Statistical test vs s* - 45% ± 6 r*', p<0.081) results: ANOVA & Duncett's multiple comparator test

[0242] Table 22: Quantification of the level of oxidation, carbonylation of proteins (%) in the dermis of skin explants treated for 24h with the IA extract (Pdo solvent) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with the topical application of urban particles (0.375qg / cm2; Fine Dust PM10-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0243] As expected, exposure of skin expiants to photo-pollution induces a significant increase in the level of oxidation and protein carbonylation in all skin compartments, including the epidermis and dermis. Treatment of skin expiants with extract IA (lot D19587, solvent Pdo) of Cananga odorata flower cake showed significant protection against stress-induced protein carbonylation in whole skin tissue (-41% ± 4 vs stress at 0.5%, p<0.001), including the epidermis (-40% ± 6 vs stress at 0.5%, p<0.001) and the dermis (-45% ± 6 vs stress at 0.5%, p<0.001).

[0244] The results for extract 2A (solvent BGP115) are recorded in the following tables 23 to 25.

[0245] [Tables23] Extract 2A (solvent 8GP115) Epidermis + Dermis Protein oxidation level % (normalized relative to the stressed condition) Treated specimen Stress (PM + UV-A) OI9548 (SSP11S) (2%) + Stress OI19545 (BGP11S) (1%) + Stress OI19548 (BGP15) (0.5%) + Stress Fi— 1 43.5 91.7 42.8 645 523 43.2 973 59.6 74.9 54.3 n=3 55.5 113.8 55.2 66.9 58.5 Mean SI wo 56 69 59 ET 4 w 12 9 6 % decrease in protein oxidation level at 0.5% vs Stress (PM + UV-A) n=1 experiment - 41% ± 6 Statistical test vs. stress: ANOVA & Dunn's multiple comparisons test

[0246] Table 23: Quantification of the level of oxidation, protein carbonylation (%) in the epidermis and dermis of skin explants treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with the topical application of urban particles (0.375qg / cm2; Fine Dust PMIO-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0247] [Tables24] Extract 2A (SGPilSj solvent) Epidermis Protein oxidation level % (normalized to stress condition) Untreated Stress (PM + WA) D19548 (BGP115) {2%} Stress D19548 (BGP115) (1%) + Stress D19S48 {BGP115J (0.5%) + Stress n=1 51.3 46.4 64.1 55.7 n=2 52.9 93.8 66.8 61.8 55.2 102.2 73.3 58.3 59.1 Mean 53 ISO 62 66 59 3 5 14 2 3 % decrease in protein carbonyl levels at 0.5% vs Stress (PM + UV-A) n=1 experiment Statistical test v - 41% t ï (*« p <d,n»l| s stress: anova & dunnetfs muitipis compariso-rss test

[0248] Table 24: Quantification of the level of oxidation, carbonylation of proteins (%) in the epidermis of skin explants treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with the topical application of urban particles (0.375qg / cm2; Fine Dust PM10-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0249] [Tables25] Extract 2A (solvent BGF115) Dermis Protein oxidation level % (normalized to stressed condition) Untreated Stress (PM + UV-A) D13548 (BGP11S) (2%) + Stress D1SS48 (BGP115) (1%) + Stress D1354S (8GP115) (0.5%) + Stress n=ln=2 40.7 43.1 49.8 100.2 93.3 106.5 59.8 68.2 65.2 653 65.(6 72.5 56.5 54.7 52.8 Mean EC 45 5 100 7 65 4 55 2 % decrease in protein carbonylation level to O,S% vs Stress (PM + UV-A) experiment Statistical test y -45%±2(***, p<0.001) stress: ANOVA & Outwett's multiple comparisons test

[0250] Table 25: Quantification of the level of oxidation, carbonylation of proteins (%) in the dermis of skin explants treated for 24h with extract 2A (solvent BGP115) of Cananga odorata flower cakes at 0.5%, 1% and 2%, then stressed with the topical application of urban particles (0.375qg / cm2; Fine Dust PM10-like; ERMCZ100) followed by UVA irradiation (6J / cm2).

[0251] As expected, exposure of skin expiants to photo-pollution induces a significant increase in the level of oxidation and protein carbonylation in all skin compartments, including the epidermis and dermis. Treatment of skin expiants with extract 2A (solvent BGP115) of Cananga odorata flower cake showed significant protection against stress-induced protein carbonylation in whole skin tissue (-41% ± 6 vs stress at 0.5%, p<0.001), including the epidermis (-41% ± 3 vs stress at 0.5%, p<0.001) and the dermis (-45% ± 2 vs stress at 0.5%, p<0.001).

[0252] Example 8: Composition Examples

[0253] Compositions have been formulated with extracts of Cananga odorata IA (solvent Pdo), 2A (solvent BGP115), and 3A (solvent SPG115) according to the invention. A serum composition is described in Table 26. A night cream composition is described in Table 27. A mask composition is described in Table 28.

[0254] SERUM

[0255] [Tables26] Phase A1 Ingredient Names DEMINERALIZED WATER Composition (Mass) 53 88 A1 BUTYLENE GLYCOL 15.00 A1 CARBOPOL ULTREZ 21 0.20 A2 ARISTOFLEXAVC 0.50 A3 GLYCERIN 12.00 A3 XANTHAN GUM 0.20 A4 DISODIUM EDTA 0.10 B EMULFREE® CBG 5.00 B DJMETHICONE 10.00 C SODIUM HYDROXIDE (10% SOL.) 0.40 D PHENOXYETHANOL, ETHYLHEXYLGLYCERIN 0.70 E Cananga odorata extract according to the invention 2.00 P UNICERT® RED 07004 J (1.0% SOL.) 0.02 TOTAL 100.00

[0256] NIGHT CREAM

[0257] [Tables27] Phase Ingredient Name Composition (% by mass) A1 ORIGINAL ORGANIC LEMON EXTRACT 15.0 A1 DEMINERALIZED WATER 31.4 A1 GLYCERIN 5.0 A2 MODIFIED POTATO STARCH 2.0 A3 EUMULGIN® SG 0.2 A3 DEMINERALIZED WATER 10.0 B: EMULIUM® MELLIFERA MB 5.0 B: DICAPRYLYL CARBONATE 15.0 B GETEARYL ALCOHOL 1.5 B UPOCR™ A SG 10.0 C EUXYL™ O12 1.2 C Cananga extract (odorants according to the invention) 2.0 C FRAGRANCE 0.2 D CITRIC ACID (10% SOL) 1.5 TOTAL 100.0

[0258] MASK

[0259] [Tables28] Phase Name of ingredients Composition (% by mass) A EMUL1UM® DELTA MB 7.0 A LABRA.F.AC™ CG MB 4.0 A LIPOCIRE^ASG 4.0 A MOD MB 2.0 A DIMETHICOE 4.0 B DEMINERALIZED WATER 45.6 B AVICEL® PC 591 3.0 C GLYCERIN 17.0 c XANTHAN GUM 0.1 D MAGNESIUM ALUMINUM SILICATE, KAOLIN 10.0 D Cananga odorafa extract according to the invention 2.0 D PARFUM 0.3 D PHENOXYETHANOL, ETHYLHEXYLGLYCERIN 10 TOTAL 100

[0260] Extracts IA (solvent Pdo), 2A (solvent BGP115) and 3A (solvent SPG115) incorporate perfectly into the compositions and provide the expected benefits as described in the preceding examples.

[0261] Extracts IA (solvent Pdo), 2A (solvent BGP115) and 3A (solvent SPG115) have the advantage of being able to be used directly by the formulator for the preparation cosmetic and / or dermatological compositions can be prepared without prior solvent removal from the extract. These extraction solvents are considered "ingredient" solvents, meaning they can be used in the composition of these cosmetic and / or dermatological compositions. They are compatible with and non-toxic to humans and the environment.

Claims

Demands

1. A process for valorizing a Cananga odorata plant waste to obtain a Cananga odorata extract, said process comprising a first solid / liquid extraction step carried out on said plant waste using an extraction solvent, followed by a second solid / liquid separation step and a third recovery step of the liquid phase constituting the extract, characterized in that the extraction solvent is anhydrous and non-polar.

2. A process according to claim 1, characterized in that the Cananga odorata plant waste consists of a fresh or dry, whole or ground cake obtained from fresh or dried Ylang Ylang flowers.

3. A process according to any one of the preceding claims characterized in that the extraction solvent comprises at least one diol comprising 3 or 4 carbon atoms, preferably at least 10% by weight relative to the total weight of the extraction solvent.

4. A process according to any one of the preceding claims characterized in that the extraction solvent is either made up of a diol comprising 3 or 4 carbon atoms, or made up of a mixture of betaine, glycerin and a diol comprising 3 or 4 carbon atoms, or made up of a mixture of sorbitol, a diol comprising 3 to 4 carbon atoms and glycerin.

5. A process according to any one of the preceding claims characterized in that the extraction solvent consists either of a mixture of betaine, glycerin and a diol comprising 3 or 4 carbon atoms in molar proportions between 1:1:3 and 1:1:7, preferably 1:1:5, or of a mixture of sorbitol, a diol comprising 3 or 4 carbon atoms, and glycerin in molar proportions between 1:1:3 and 1:1:7, preferably 1:1:5, the diol comprising 3 or 4 carbon atoms being preferably propanediol, the extraction solvent being preferably a NaDES solvent.

6. Extract of Cananga odorata obtained according to the process according to any one of claims 1 to 5.

7. Extract of Cananga odorata characterized in that it comprises an allergenic compound content of less than 0.1% by weight relative to to the total weight of the extract, the allergenic compounds being defined according to EU regulation 2023 / 1545.

8. Extract of Cananga odorata according to any one of claims 6 or 7, characterized in that it is non-oily and contains less than 20% by weight of water.

9. Non-therapeutic use of the Cananga odorata extract obtained according to the process according to any one of claims 1 to 5, or of the extract according to any one of claims 6 to 8, or of a composition comprising it, for the cosmetic treatment of the skin and / or mucous membranes, for the improvement of the appearance of the skin and / or mucous membranes, for the improvement of skin resistance and / or elasticity, and / or for the treatment or prevention of skin aging, wrinkles, or skin laxity.

10. Non-therapeutic use of the Cananga odorata extract obtained according to the process according to any one of claims 1 to 5, or of the extract according to any one of claims 6 to 8, or of a composition comprising it, to combat skin aging and / or protect the skin from external aggressions.

11. Non-therapeutic use of the Cananga odorata extract obtained according to the process according to any one of claims 1 to 5, or of the extract according to any one of claims 6 to 8, or of a composition comprising it, to increase skin hydration, and / or strengthen the skin barrier function, and / or improve skin microrelief, and / or improve skin radiance and tone.

12. Extract of Cananga odorata obtained according to the process according to any one of claims 1 to 5, or extract according to any one of claims 6 to 8, or composition comprising thereon, for use to increase the transcriptomic expression of antioxidant enzymes and / or to stimulate the Nrf2 antioxidant pathway, and / or to increase protection against stress-induced protein carbonylation in skin tissue.

13. Cosmetic and / or dermatological composition comprising at least the extract of Cananga odorata obtained according to the process according to any one of claims 1 to 5, or the extract according to any one of claims 6 to 8.

14. Composition according to claim 13, characterized in that the extract of Cananga odorata represents between 0.1% and 10% by weight of said composition.

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