Active ingredient comprising cyclitols obtained from a Ginkgo biloba extract and its cosmetic uses

Abstract

The present invention relates to an active ingredient comprising cyclitols obtained from a Ginkgo biloba extract, a cosmetic composition comprising it and its anti-aging cosmetic uses.

Description

Title of the invention: Active ingredient comprising cyclitols obtained from a Ginkgo biloba extract and its cosmetic uses. Technical field

[0001] The invention relates to an active ingredient comprising cyclitols obtained from a Ginkgo biloba extract, a cosmetic composition comprising it and its anti-aging cosmetic uses.

[0002] State of the art

[0003] Over the past few decades, research has made major advances in understanding the mechanisms related to aging. It is now well established that the entry of cells into senescence is one of the central factors in tissue aging.

[0004] In the skin, the more a fibroblast divides, the more it ages. However, fibroblasts have a limited number of divisions which, when exhausted, triggers the entry of the cell into senescence. They can no longer divide but remain metabolically active and capable of influencing the fate of the tissue. They then develop a particular phenotype allowing them to communicate with cells present in their environment (fibroblasts, immune cells, keratinocytes, etc.). This communication notably involves the secretion of a significant quantity of inflammatory molecules grouped under the term SASP (senescence-associated secretory phenotype), including interleukin 6 (IL-6), and metalloproteinases 1 and 3 (MMP-1 and MMP-3).

[0005] In the long term, the accumulation of these molecules leads to an alteration in the functionality of the surrounding cells. The ability of fibroblasts to synthesize the components of the extracellular matrix also deteriorates. The dermis loses density, and the epidermis struggles to renew itself, resulting in the appearance of wrinkles, a loss of elasticity, and uneven skin tone.

[0006] Immune cells are capable of efficiently identifying and eliminating these senescent cells, preventing them from causing damage and thus premature aging of tissues. In young skin, senescent cells, via SASPs, attract immune cells, which recognize them through specific ligands belonging to the MIC (MHC Class 1 Polypeptide-Related Sequence) family, notably MICA and MICB, in order to eliminate them.

[0007] Among the immune cells responsible for eliminating senescent cells, NK (Natural Killer) cells are the best known. They possess specific cell death activating receptors (NKG2D receptors) on their surface. MICA / B ligands. Through the action of immune cells on senescent cells, the skin is preserved from premature aging.

[0008] In aged skin, senescent fibroblasts have developed strategies that allow them to deceive the immune system, leading to their accumulation in the dermis and promoting skin aging. To achieve this, senescent fibroblasts have developed a phenotype that allows them to cleave the specific ligand (MICA / MICB), but also to increase the expression on their surface of a "self" protein, called HLA-E (class I histocompatibility antigen, alpha chain E), which is specific to the NKG2A receptor of NK cells, enabling them to recognize senescent fibroblasts as healthy cells to be preserved.

[0009] In order to combat this phenomenon, the cosmetics industry initially sought active ingredients capable of delaying the entry of cells into senescence, and then focused on targeting these senescent cells. However, the molecules capable of eliminating senescent cells mostly exhibit significant toxicity due to their lack of specificity.

[0010] There is therefore a need for a new cosmetic active ingredient that can specifically and non-toxically target senescent cells in order to eliminate them. Summary of the invention

[0011] This is the objective of the present invention. In this context, the inventor became interested in the mechanism of senolysis, that is to say the elimination of senescent fibroblasts from the skin, and observed the harmful effects of the prolonged spread of SASP on their environment.

[0012] The inventor then discovered a new active ingredient, obtained from Ginkgo biloba, that specifically targets senescent fibroblasts, restoring senolysis by immune cells, thus preserving the skin from natural aging. Through the action of this new active ingredient, the density and elasticity of the dermis are improved, wrinkles are reduced, and the complexion's radiance is enhanced.

[0013] Ginkgo biloba is a tree with exceptional longevity, as it can live for over 1000 years, and even up to 3000 years for some varieties. Beyond this remarkable longevity, its immune system, surprisingly, does not weaken over time, the foliage remains as exuberant as ever, trunk growth does not slow down, and the seeds remain highly fertile.

[0014] Given the undeniable qualities of Ginkgo biloba, cosmetic active ingredients based on Ginkgo biloba are already known to combat aging; however, unlike the present invention, these aim to delay the entry of cells into senescence as much as possible by preserving fibroblasts.

[0015] The inventor has identified a particular extract of Ginkgo biloba enriched in cyclitols, this which enabled him to develop a new anti-aging cosmetic active ingredient, allowing the restoration of the natural senolysis of fibroblasts, restoring the immune system's ability to eliminate senescent cells and thus meeting the needs of the present invention.

[0016] In particular, the extract according to the invention makes it possible to: - significantly reduce the secretion of central inflammatory molecules, in particular the secretion of IL-6, MMP-1 and MMP-3; - significantly limit the cleavage of the immunogenic ligand MICA; and - to improve the lysis of refractory senescent fibroblasts by NK cells compared to untreated cells.

[0017] Thus, the invention relates to a cosmetic active ingredient comprising at least one extract of Ginkgo biloba, said extract comprising cyclitols.

[0018] Preferably, said extract is obtained from the leaves of Ginkgo biloba, more preferably from the young leaves, that is to say the green leaves, in particular before they take on their characteristic golden yellow tint.

[0019] In order to further enrich the cyclitol content in the extract, by allowing the release of the less accessible cyclitols, hydrolysis, preferably enzymatic hydrolysis, is carried out. Thus, the extract according to the invention is preferably a hydrolysate, more preferably an enzymatic hydrolysate.

[0020] According to a particular aspect of the invention, the cyclitol-rich extract is a filtered hydrolysate, unlike an extract obtained from maceration or infusion. The filtration performed is frontal filtration, which can be carried out by screening or sieving using a membrane filter. This filtration can be performed by reverse osmosis, microfiltration, or ultrafiltration, depending on the size of the molecules that those skilled in the art wish to obtain.

[0021] The extract according to the invention is particularly rich in cyclitols, also, the cyclitols preferably represent at least 5% by weight of the total dry weight of the extract.

[0022] The extract according to the invention may also comprise other molecules, preferably including carbohydrates. The carbohydrates may be of different types, in different forms, and be more or less polymerized. Thus, the extract according to the invention comprises monosaccharides.

[0023] Preferably, the extract according to the invention also comprises carbohydrates with a degree of polymerization ranging from 2 to 34. The degree of polymerization (DP) defines the length of a polymer chain, that is, the number of monomer units present in the chain. A degree of polymerization of 2, or DP2, corresponds to the presence of two monomers. More preferably, the monosaccharides represent 80% by weight of the total weight of the carbohydrates, and / or the carbo hydrates having a degree of polymerization between 2 and 34 represent 20% by weight of the total weight of carbohydrates.

[0024] The active ingredient according to the invention advantageously comprises at least one excipient selected from a preservative, an antioxidant, a stabilizer, a spray-drying agent, and / or a combination thereof. Furthermore, the active ingredient according to the invention preferably consists of Ginkgo biloba extract and at least one excipient.

[0025] According to a particularly preferred object, the cosmetic active ingredient according to the invention is capable of being obtained by a process comprising the following steps: a. solubilization of at least 50 g / L of Ginkgo biloba in water, b. Enzymatic hydrolysis, c. Separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, d. Enzymatic inactivation, and e. possibly concentration and sterilizing filtration.

[0026] Also, according to another aspect, the invention also relates to a preparation process comprising the above steps, to obtain the active ingredient according to the invention.

[0027] According to another aspect, the invention also relates to anti-aging cosmetic uses; preferably, said anti-aging cosmetic uses of the active ingredient according to the invention aim to reduce wrinkles and / or improve skin tone evenness, and / or revive skin radiance. To achieve this, the active ingredient according to the invention acts on senescent fibroblasts by preventing them from deceiving the immune system and preventing their accumulation in the dermis.

[0028] Also, the invention also relates to the use of the active principle to restore the natural elimination of senescent fibroblasts, or senolysis, by immune cells.

[0029] On the other hand, said active principle according to the invention is intended for topical application on the skin, preferably to increase cell renewal and / or dermal density, and / or dermal elasticity.

[0030] The invention also relates to a cosmetic composition for topical application comprising at least the active ingredient according to any of the preceding embodiments and a physiologically acceptable medium, preferably the active ingredient represents at least 0.1% by weight of the total weight of the composition.

[0031] Other features and advantages will become apparent from the detailed description of the invention, the examples and figures that will follow. Detailed description of the invention

[0032] Definition

[0033] For the purposes of this invention, "cosmetic active ingredient" means an extract comprising at least one molecule, preferably a set of molecules having a cosmetic effect on the skin when applied topically.

[0034] For the purposes of this invention, "naturally derived ingredient" means a cosmetic ingredient obtained through defined chemical and / or biological processes aimed at chemically modifying it. These chemical modifications are intentional, and the structures of the resulting molecules are therefore no longer identical to molecules found in nature.

[0035] For the purposes of this invention, "Ginkgo biloba hydrolysate" means an active ingredient derived from a tree of the species Ginkgo biloba, preferably from Ginkgo biloba leaves, obtained by a process comprising at least one hydrolysis step of Ginkgo biloba. The term "Ginkgo biloba hydrolysate" excludes molecules produced solely by fermentation of Ginkgo biloba by a microorganism or by infusion / decoction / maceration of Ginkgo biloba.

[0036] For the purposes of this invention, "filtered hydrolysate of Ginkgo biloba" means an active ingredient derived from a tree of the species Ginkgo biloba, preferably from the leaves of Ginkgo biloba, obtained by a process comprising at least one step of hydrolysis and filtration of Ginkgo biloba, more preferably the filtration step being carried out during the step of separating the soluble and insoluble phases after the hydrolysis step. The term "filtered hydrolysate of Ginkgo biloba" excludes molecules produced solely by fermentation of Ginkgo biloba by a microorganism or by infusion / decoction / maceration of Ginkgo biloba.

[0037] For the purposes of this invention, "young Ginkgo biloba leaves" means green Ginkgo biloba leaves obtained / harvested before they turn yellow, in particular before they develop their characteristic golden yellow tint.

[0038] Active ingredient according to the invention

[0039] The present invention therefore relates to a cosmetic active ingredient comprising at least one extract of Ginkgo biloba, characterized in that the extract comprises cyclitols. The cosmetic active ingredient according to the invention is a natural-derived ingredient within the meaning of ISO 16128.

[0040] Ginkgo biloba is the oldest known tree species on Earth. It belongs to the Ginkgoaceae family and is also known as the maidenhair tree or silver apricot tree. Native to Asia, this tree contains powerful antioxidants such as flavonoids and terpenes, which are particularly known to slow down cell aging. It is also known to improve circulation. It is beneficial for blood circulation and overall well-being. Ginkgo biloba is therefore commonly used for its numerous health benefits for humans.

[0041] Thanks to its properties and its particularly rich cyclitol content, the extract according to the invention has demonstrated cross-layered efficacy on the dermis and epidermis of mature skin. This ingredient, derived from natural sources and obtained from Ginkgo biloba, specifically targets senescent fibroblasts, restoring the natural senolysis of these fibroblasts and re-establishing the immune system's ability to eliminate senescent cells. This new active ingredient boosts epidermal renewal, improves dermal density, reduces wrinkles, helps the skin regain its elasticity, and enhances the complexion's radiance.

[0042] The cyclitols present in the extract according to the invention are cyclic polyols, which contain hydroxyl groups and are particularly stable under ordinary physiological conditions due to their cyclic structure. They are considered osmoprotectants, meaning they can act as osmolytes in plants, enabling the organism to survive significant osmotic stress. This family of molecules includes, in particular, pinnitol, myo-inositol, pinpollitol, and ciceritol. This type of molecule is present in various plants, such as Ginkgo biloba and the ash tree (Fraxinus excelsior), at concentrations and in proportions that vary depending on the species.

[0043] Also, the invention relates to an active ingredient comprising at least one extract of Ginkgo biloba rich in cyclitols, as well as to its cosmetic uses, preferably to reduce wrinkles and / or improve the evenness of the complexion, and / or revive the radiance of the complexion.

[0044] In order to develop such an ingredient derived from nature (according to the definitions of ISO 16128) enriched in cyclitols, the extract included in the active principle according to the invention is preferably obtained from Ginkgo biloba leaves, more preferably from young Ginkgo biloba leaves.

[0045] In order to increase the cyclitol content, the extract is preferably a hydrolysate, more preferably a filtered hydrolysate, even more preferably a filtered enzymatic hydrolysate, which allows the release of the cyclitols that are difficult to access.

[0046] Also, according to a particularly preferred object of the invention, the extract comprises at least 5% cyclitols, by weight of the total dry weight of the extract.

[0047] The quantification of cyclitols can be determined by ion chromatography. Calibration solutions are prepared from a standard (Pinnitol) with a concentration between 2 and 10 mg / L. The extract according to the invention is first diluted in water so that the cyclitol content corresponds to the calibration range. Finally, the quantity of cyclitols in the samples is determined using a calibration curve.

[0048] The dry matter content of the extract can be determined by weighing the residues obtained from drying the extract according to the invention at 105°C in an oven until a constant weight is obtained. Preferably, the extract according to the invention in liquid form has a dry matter content of between 20 g / l and 50 g / l.

[0049] The inventor continued the characterization of the extract according to the invention, which preferentially comprises carbohydrates. The carbohydrate content can notably be determined using the DUBOIS method (Dubois M. et al., Analytical chemistry, 28, 3, 350-356, 1956).

[0050] The molar masses of the carbohydrates were also determined. Furthermore, the carbohydrates present in the extract according to the invention preferably have a degree of polymerization between 1 and 34.

[0051] In a particularly preferred manner, monosaccharides having a degree of polymerization of 1, i.e. a molar mass less than 180 Da, represent 80% by weight of the total weight of carbohydrates, and / or carbohydrates having a degree of polymerization between 2 and 34 and therefore a molar mass between 360 and 6120 Da represent at least 20% by weight of the total weight of carbohydrates.

[0052] The extract according to the invention is also composed of other molecules, such as proteins, uronic acids, organic acids and mineral ash.

[0053] The distribution and quantity of the protein fraction can be determined by the determination of total nitrogen according to the Kjeldahl method (reference: Official method of analysis of the AOC, 12th ed. W. Horwitz, ED, New York, 15-60, 1975). Preferably, the extract comprises 2% peptides by weight of dry matter of the extract according to the Kjeldahl method, having a molecular mass of less than 2,000 Da.

[0054] The inventor also determined the mineral ash content. The mineral ash content can be determined by weighing the residues from the incineration of samples of the active ingredient according to the invention at 550°C in an electric muffle furnace. Preferably, the active ingredient according to the invention has a mineral ash content of between 25% and 40% by weight of the dry matter of the active ingredient.

[0055] The inventor also determined the organic acid content. This was determined by ion chromatography (DIONEX). Preferably, the active ingredient according to the invention has an organic acid content of between 20 and 40% by weight of dry matter.

[0056] The inventor has finally determined the uronic acid content. The reaction between uronic acids and sodium tetraborate, in the presence of metahydroxydiphenyl, produces a pink color that allows for quantification using a spectrophotometer at 250 nm. The active ingredient according to the invention contains between 5% and 13% uronic acids by weight. of dry materials.

[0057] The active ingredient according to the invention may also include other ingredients added by those skilled in the art, in particular excipients. The active ingredient according to the invention preferably comprises at least one excipient selected from a preservative, an antioxidant, a stabilizer, a spray-on support, and combinations thereof. Such excipients stabilize, preserve, and prevent the degradation of the extract, in particular of the molecules contained within the extract, such as the cyclitols responsible for the anti-aging cosmetic efficacy of said active ingredient.

[0058] The cosmetic active ingredient according to the invention can be in different forms, preferably in liquid form, in solid form or in film form.

[0059] When in liquid form, the active ingredient according to the invention consists exclusively of Ginkgo biloba extract accompanied by excipients, preferably stabilizer and / or preservative, and / or antioxidant.

[0060] The extract in liquid form is preferably a clear aqueous solution with a faint odor and a light yellow to yellow color. However, it can be more colored and / or decolorized by any process known to those skilled in the art.

[0061] When in solid form, the active ingredient according to the invention preferably consists of Ginkgo biloba extract as previously described and at least one excipient such as a spray-drying support selected from maltodextrin, gum arabic, or soy lecithin. It may also include a stabilizer and / or preservative, and / or an antioxidant in addition to the support. In a particularly suitable embodiment, the extract represents at least 10% by weight of the active ingredient and the support at most 90% by weight of the active ingredient.

[0062] In the case of a solid form in which the active ingredient is associated with a support, the levels of proteins, sugars, cyclitols, uronic acids, organic acids and mineral ash in the active ingredient are modified, the support generally being made up mainly of sugars.

[0063] The active ingredient according to the invention can also be presented in the form of a film. In this case, the Ginkgo biloba extract preferably represents at least 0.1% by weight of the film.

[0064] When presented in film form, the active ingredient comprises: - at least the Ginkgo biloba extract according to the invention, - at least one mineral charge, and - at least one polymer of natural origin, and - at least one plasticizer, and - at least one surfactant.

[0065] The polymer of natural origin can be chosen from: Pectin, Tamarind gum, Alginate, Pullulan, Psyllium, Xanthan gum, Guar gum, Tara gum, Carob gum, Agar, Gum arabic, Gellan gum, Dextran, Carrageenan, Cellulose, Konjac and Chitosan.

[0066] The plasticizer can be chosen from: Glycerol, Sorbitol, Sucrose, Erythritol, Urea, Propylene glycol and Butylene glycol.

[0067] The mineral filler can be chosen from: Calcium carbonate, green clay, kaolin, perlite, talc, magnesium silicate, mica, diatomaceous sericite, silica, calcium sulfate, calcium chloride, potassium chloride, iron oxide and zinc oxide.

[0068] The active ingredient in film form may further comprise a pigment for coloring said film.

[0069] According to a particularly preferred embodiment, the extract of the cosmetic active ingredient according to the invention can be obtained by an extraction process comprising the following steps: a. solubilization of at least 50 g / L of Ginkgo biloba in water, b. Enzymatic hydrolysis, c. Separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, d. Enzymatic inactivation, and e. possibly concentration and sterilizing filtration.

[0070] Cosmetic composition according to the invention

[0071] The active ingredient according to the invention can optionally be incorporated into a cosmetic composition, in particular a composition comprising at least 0.1% by weight of said active ingredient and a physiologically acceptable medium, preferably a cosmetically acceptable medium.

[0072] These compositions may be in the form of oil-in-water emulsions, water-in-oil emulsions, multiple emulsions (Water / Oil / Water or Oil / Water / Oil), which may be microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersions, aqueous gels, powders, foundation, or films. They may be more or less fluid and have the appearance of creams, emulsions, gels, masks, or any other type of cosmetic product for the care of healthy skin.

[0073] These may be compositions comprising at least 0.1% of the liquid active ingredient according to the invention, preferably between 0.1 and 10%.

[0074] These compositions comprise, in addition to the active ingredient, a physiologically acceptable medium such as a cosmetically acceptable medium, that is to say, one which does not cause any discomfort to the user such as redness, tightness or tingling.

[0075] The compositions according to the invention may contain as an adjuvant at least one composed chosen from: - oils, which can be chosen in particular from among silicone oils, linear or cyclic, volatile or non-volatile, - waxes, such as ozokerite, polyethylene wax, beeswax or camauba wax, - silicone elastomers, - surfactants, preferably emulsifying, whether non-ionic, anionic, cationic or amphoteric, - co-surfactants, such as linear fatty alcohols, - thickeners and / or gelling agents, - humectants, such as polyols like glycerin, - colorings, preservatives, fillers, - the tensors, - the sequestrators, - perfumes, - and their mixtures, without this list being exhaustive.

[0076] Examples of such adjuvants are cited in particular in the CTFA Dictionary (International Cosmetic Ingredient Dictionary and Handbook published by the Personal Care Product Council).

[0077] Of course, a person skilled in the art will take care to choose any additional compounds, active or inactive, and their quantity, in such a way that the advantageous properties of the mixture are not, or substantially not, altered by the envisaged addition.

[0078] Extraction method according to the invention

[0079] The extract constituting or contained in the active ingredient according to the invention can be obtained by any known means allowing the production of an extract rich in cyclitols.

[0080] According to a preferred embodiment, the extraction process includes at least one solubilization step of Ginkgo biloba leaves, a hydrolysis step, preferably an enzymatic hydrolysis, to increase the cyclitol content and a filtration step.

[0081] Prior to the extraction process itself, Ginkgo biloba leaves are harvested from Ginkgo biloba cultivated using environmentally friendly methods that combine intercropping, permanent ground cover, the judicious use of plant-friendly green manure, and the planting of wild hedgerows. This cultivation method protects the Ginkgo biloba crops from external pesticide contamination. Preferably, the leaves are harvested manually and selectively so as to collect only the young green leaves before they acquire their characteristic golden-yellow color.

[0082] According to a particularly suitable embodiment, the active ingredient according to the invention is obtained by carrying out the following steps: a. Solubilization of at least 50 g / L of Ginkgo biloba in water, preferably green Ginkgo biloba leaves, b. Hydrolysis, preferably enzymatic hydrolysis, c. Separation of soluble and insoluble phases by filtration and recovery of the soluble phase, d. Enzyme inactivation, and e. possibly concentration and sterilizing filtration.

[0083] The hydrolysis conditions are chosen to obtain an extract enriched in cyclitols, preferably at least 5% by weight of the total dry matter weight.

[0084] The separation of the soluble and insoluble phases is carried out by filtration. The separation of the soluble and insoluble phases is carried out to recover the soluble phase containing, among other things, cyclitols, i.e., the filtered hydrolysate.

[0085] Optionally, an additional heat treatment step may be carried out to inactivate the enzyme used. This inactivation is then carried out according to the recommendations of the supplier of the enzyme used.

[0086] Optionally, the process includes a filtration step after recovery of the soluble phase to remove any remaining suspended particles. This filtration step purifies the recovered soluble phase and is performed to select the active molecules.

[0087] The hydrolysate obtained at this stage can optionally be further concentrated and / or purified, preferably by successive ultrafiltration steps through membranes of different porosity, retaining the active molecules at each step and / or by a chromatographic method.

[0088] The hydrolysate obtained after hydrolysis and filtration, before or after concentration and sterilizing filtration, is a hydrolysate of Ginkgo biloba, and constitutes a first form of the active principle according to the invention, then being in liquid form.

[0089] The resulting hydrolysate can then be dried and, with or without an excipient or support, to obtain, for example, a solid form. This phase can be carried out by performing the following steps: - an atomization support, preferably maltodextrin, is added to the Ginkgo biloba hydrolysate, at most 90% (by mass / volume); - this solution is then concentrated under vacuum; - any bacteria that may be present are eliminated by heat treatment; - Atomization allows us to obtain a powder.

[0090] The steps of the processes described above, taken individually, are usual in the field of extracting active ingredients from natural raw materials and the person skilled in the art is able to adjust the reaction parameters based on their general knowledge.

[0091] Cosmetic use

[0092] The active ingredient according to the invention or the composition according to the invention is intended to be used on healthy skin, in particular mature healthy skin, especially as an anti-aging cosmetic active ingredient.

[0093] Preferably, the active ingredient according to the invention is intended for topical application to the skin to reduce wrinkles and / or improve the evenness of the complexion, and / or revive the radiance of the complexion.

[0094] To achieve this, the active principle according to the invention restores the natural elimination of senescent fibroblasts, or senolysis, by immune cells.

[0095] The active ingredient thus increases cell renewal and / or dermal density, and / or dermal elasticity.

[0096] The extract according to the invention protects the cellular environment from aging. Although few senescent cells are present in the tissue, a single senescent cell can affect many others due to the paracrine signaling that exists between different cell types. Thus, senescent fibroblasts that escape elimination by NK cells accumulate and secrete SASP inflammatory markers in abundance and without interruption, accelerating skin aging.

[0097] Furthermore, fibroblasts produce certain structural components of the extracellular matrix (ECM), such as collagen. However, the accumulation of senescent fibroblasts leads to an imbalance between matrix synthesis and degradation. On the one hand, the prolonged secretion of MMP-type collagenases, particularly MMP-1 and MMP-3, results in the degradation of the existing ECM. On the other hand, the SASP (synthetic angiotensin systemic sclerosis) of senescent fibroblasts also disrupts the metabolism of surrounding fibroblasts, which then struggle to renew the matrix. The dense and firm network of collagen fibers becomes disorganized, ultimately leading to a loss of the skin's biomechanical properties and the formation of wrinkles.

[0098] However, the extract according to the invention limits these harmful effects, resulting in a significant improvement in dermal density and elasticity of mature skin. Consequently, the extract according to the invention significantly reduces wrinkles.

[0099] Senescent fibroblasts persisting in the tissue also weaken epidermal-dermal interactions, thus leading to an alteration in the epidermal renewal process. Excessive secretion of inflammatory molecules from the SASP induces the degradation of protein structures that anchor the epidermis to the dermis. This secretome, which is harmful to the tissue, notably causes the loss of an essential marker for maintaining epidermal and skin homeostasis: collagen XVII.

[0100] The extract according to the invention also limits the deleterious effects of senescent fibroblasts on the production of collagen XVII by keratinocytes, resulting in a significant improvement in cell renewal on mature skin and therefore a more even and revived complexion.

[0101] Thus, the extract according to the invention, through its cross-cutting action, boosts epidermal renewal, improves dermal density, and enhances its elasticity, reducing wrinkles, and also enhances the radiance and evenness of the complexion. The invention also relates to the use of the active ingredient to restore the natural elimination of senescent fibroblasts, or senolysis, by immune cells.

[0102] The invention is now illustrated by non-limiting examples of compositions according to the invention and by results. Examples

[0103] Example 1: Active ingredient according to the invention (AI)

[0104] The active ingredient of Example 1 is obtained from the leaves of Ginkgo biloba. The active ingredient of Example 1 is obtained by the following process: - Solubilization of at least 50 g / L of Ginkgo biloba leaves in water, - Enzymatic hydrolysis, - Separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, i.e., the filtered hydrolysate, - Thermal treatment - Purification and concentration of the soluble phase, - Filtration and sterilizing filtration.

[0105] The active ingredient obtained has the following characteristics: - Dry matter content = 31.4 g / L - Cyclitol content = 2.4 g / L, or 8%, by weight relative to dry matter, - Total Sugar Content (according to the Dubois method) = 6.7 g / L, or 21%, in weight relative to dry matter, - Galacturonic acid content = 2g / l, or 6%, by weight relative to dry matter, - Clear, yellow liquid, faint odor

[0106] Example 2: Active ingredient outside the invention (HI1)

[0107] The active ingredient of Example 2 is obtained from Ginkgo biloba leaves. The active ingredient of Example 2 is obtained by the following process: - Solubilization of at least 50 g / L of Ginkgo biloba in water, - Separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, - Thermal treatment - Purification and concentration of the soluble phase, - Filtration and sterilizing filtration.

[0108] The active ingredient obtained has the following characteristics: - Dry matter content = 28.8 g / L - Cyclitol content = 1.74 g / L, or 6%, by weight relative to dry matter, - Total Sugar Content (according to the Dubois method) = 6.4 g / L, or 22%, by weight relative to dry matter, - Galacturonic acid content = 0.84g / l, or 3%, by weight relative to dry matter, - Clear, very pale yellow liquid, faint odor

[0109] Example 3: Active ingredient outside the scope of invention (fHI2)

[0110] The active ingredient of Example 3 is obtained from ash of the species Fraxinus excelsior. This active ingredient was produced from a plant species different from Ginkgo biloba but rich in cyclitols, according to the process described in Example 1.

[0111] The active ingredient obtained has the following characteristics: - Dry matter content = 22.6 g / L - Cyclitol content = 1.53 g / L, or 7%, by weight relative to dry matter, - Total Sugar Content (according to the Dubois method) = 5.8 g / L, or 26%, by weight relative to dry matter, - Galacturonic acid content = 2.4g / l, or 11%, by weight relative to dry matter, - Clear, yellow liquid, faint odor

[0112] Example 4: Composition according to the invention

[0113] An example of a formulation of a smooth cream comprising the active ingredient according to the invention is shown in Table 1 below.

[0114] [Tables 1] Ingredients % Al Water qs 100 Preservative qs Butylene Glycol 3.00 A2 Glycerin 1.00 Xanthan Gum 0.20 B PEG-100 Stearate & Glyceryl Stearate 3.00 Myristyl Alcohol & Myristyl Glucoside 2.00 Cetearyl Alcohol 2.00 Hydrogenated Rapeseed Oil 3.00 Ethylhexyl Polyhydroxy Stearate 2.00 Isodecyl Neopentanoate 8.00 PPG-3 Benzyl Ether Myristate 8.00 Dimethicone 2.00 C Polyacrylate-13 & Polyisobutene & Polysorbate 20 1.00 D Active Ingredient 2.00

[0115] The composition of example 4 can in particular be obtained by the following process: a. Add A2 to Al under moderate stirring, stir until homogeneous and heat to 80°C. b. Place B under stirring and heat to 80°C. c. Emulsify B in A under shear agitation for 10 minutes. d. At 30-40°C, under moderate stirring, add C then D.

[0116] The composition then takes the form of a white and glossy emulsion having a pH at 1 month equal to 5.4, a viscosity (C / 5rpm) equal to 118,000 cP.

[0117] Example 5: Composition according to the invention

[0118] An example of a formulation of a global anti-aging cream comprising the active ingredient according to the invention is shown in Table 2 below.

[0119] [Tables2] Ingredients % Al Purified Water qs 100 Preservative qs Butylene Glycol 3.00 A2 Glycerin 2.00 (Water) & Polyglycerin-3 4.00 Garcinia Indica Seed Butter 2.00 Oleyl Erucate 6.00 Diisopropyl Adipate 7.00 Prunus Amygdalus Dulcis (Sweet Almond) Oil 3.00 Dimethicone 1.50 Tocopherol & Helianthus Annuus (Sunflower) Seed Oil 0.05 C Hydroxyethyl Acrylate / Sodium Acryloyldimethyl Taurate Copolymer & Polyisobutene & PEG-7 Trimethylolpropane Coconut Ether 1.50 D ACTIVE INGREDIENT INVENTION 2.00

[0120] The composition of example 5 can in particular be obtained by the following process: a. Add A2 to Al under moderate stirring, stir until homogeneous and heat to 80°C. b. Place B under stirring and heat to 80°C. c. Emulsify B in A under shear agitation for 10 minutes. d. At 30-40°C under moderate stirring, add C then D.

[0121] The composition then takes the form of a soft and glossy off-white emulsion having a pH at 1 month of 6.1, a viscosity (C / 5rpm) of 48,600 cP.

[0122] Example 6: Composition according to the invention

[0123] An example of the formulation of an anti-wrinkle serum comprising the active ingredient according to the invention is shown in Table 3 below.

[0124] [Tables3] Ingredients % A1 Purified Water q.s. 100 Butylene Glycol 4.00 A2 Glycerin 2.00 Xanthan Gum 0.15 B Polyacrylate Crosspolymer-6 0.50 Polyacrylamide & C13-14 Isoparaffin & Laureth-7 2.00 Dimethicone 15.00 Dimethicone & Dimethicone Crosspolymer 7.00 C Preservative q.s. D CI 77891 (Titanium Dioxide) & Mica & CI 77491 (Iron Oxides) 0.50 E Active Ingredient 4.00 F Citric Acid Solution 10% q.s. pH

[0125] The composition of example 6 can in particular be obtained by the following process: a. Add A2 to Al under moderate stirring, stir until homogeneous, b. Homogenize B under stirring. c. Add B to A while stirring vigorously. Shake for 20 minutes. d. Under moderate stirring, add C then D and E. e. Adjust the pH to 4.8–5.2 with F

[0126] The composition is then presented in the form of a thick, shiny, pearly coral gel-cream having a pH at 1 month equal to 4.9, a viscosity (C / 5rpm) equal to 42,200 cP.

[0127] Example 7: Composition according to the invention

[0128] An example of a formulation of a smoothing foundation comprising the active ingredient according to the invention is shown in Table 4 below.

[0129] [Tables4] Ingredients % A Propanediol Dicaprylate 6.00 Magnesium Stearate 0.80 Polyglyceryl-6 Polyricinoleate & Polyglyceryl-3 Diisostearate & Disteardimonium Hectorite 6.00 Undecane & Tridecane 7.00 Tocopherol & Helianthus Annuus (Sunflower) Seed Oil 0.10 B Pentaerythrityl Tetraisostearate 2.00 Isocetyl Stearoyl Stearate 5.00 Pigments 8.15 C Silica 3.00 DI Purified Water qs 100 Sodium Gluconate 0.20 Preservative qs D2 Anhydrous Citric Acid 0.05 Glycerin 3.00 Pentylene Glycol 3.00 Sodium Chloride 1.00 D3 Active Ingredient 2.50

[0130] The composition of example 7 can in particular be obtained by the following process: a. Pass the mixture B through the three-cylinder. b. Homogenize A using a deflocculator then, under vigorous stirring, add B then C. c. Mix DI until solubilized. Add the MP from D2 and D3 while stirring. d. Slowly add D to ABC while stirring vigorously. Stir for 10 minutes then homogenize for 2 minutes in the rotor / stator.

[0131] The composition then takes the form of a soft and glossy light beige emulsion having a viscosity (C / 5rpm) equal to 19,500 cP.

[0132] In vitro evaluation of the efficacy of the active ingredient according to the invention

[0133] Test 1 - Effect of the active ingredient according to the invention on the secretome of immunoevasive senescent fibroblasts.

[0134] The objective of this study is to evaluate the ability of the active ingredient according to the invention (AI) to limit the secretion of S ASP proteins, in particular MMP-1, MMP-3 and 11-6, by immunoevasive senescent fibroblasts in order to limit inflammation as well as degradation of the cutaneous microenvironment.

[0135] Normal and senescent human fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2. On day 3, the immunoevasive phenotype is activated. Then, the senescent human fibroblasts are stressed daily to obtain a population of immunoevasive senescent fibroblasts. At the end of each stress cycle, the cells are treated or not with the active ingredient according to the invention at 0.25% and 0.50% (v / v). The secretion of the three SASP proteins (MMP-1, MMP-3, IL-6) is then evaluated by ELISA assay.

[0136] The results on the synthesis of MMP-1, MMP-3 and IL-6 by immunoevasive senescent fibroblasts are presented in Table 5 below.

[0137] [Tables5] MMP-1 Synthesis (ng / mg protein) Capacity to Limit MMP-1 Synthesis (%) MMP-3 Synthesis (ng / mg protein) Capacity to Limit MMP-3 Synthesis (%) IL-6 Synthesis (pg / mg protein) Capacity to Limit IL-6 Synthesis (%) Normal Fibroblasts Control 639 263 118 Senescent Fibroblasts Control 1597 714 186 Immunoevasive Senescent Fibroblasts Control 3299 2451 264 PA 0.25% 2691 +36 2168 +16 247 +22 PA 0.50% 2522 +46 1967 +28 196 +87 HI1 0.50% 3231 +4 2434 +1 - - HI2 0.50% 3197 +6 2434 +1 - -

[0138] Compared to senescent fibroblasts, immunoevasive senescent fibroblasts secrete significantly more MMP-1, MMP-3 and IL-6.

[0139] Tested at 0.5% on immunoevasive senescent human fibroblasts, the active ingredient according to the invention significantly limits the synthesis of MMP-1, MMP-3, and IL-6 of respectively 46%, 28% and 87%, thus limiting the secretion of deregulated SASP proteins. Whereas the two extracts outside the scope of this invention have no effect on the synthesis of MMP-1 and MMP-3.

[0140] Test 2 - Effect of the active ingredient according to the invention on the cleavage of the MICA ligand by immunoevasive senescent fibroblasts.

[0141] The objective of this study is to evaluate the ability of the active ingredient according to the invention (AI) to limit the cleavage of the immunogenic ligand MICA, responsible for the protein signal recognized by immune system cells allowing the elimination of senescent fibroblasts.

[0142] The protocol is identical to that of trial 1 except that the rate of soluble cleaved MICA in the culture medium is evaluated by ELISA assay.

[0143] The results are presented in Table 6 below.

[0144] [Tableauxô] MICA cleaved rate (ng / mg protein) Capacity to limit MICA cleavage (%) Normal fibroblasts Control 17 Senescent fibroblasts Control 19 Immunoevasive senescent fibroblasts Control 70 PA 0.25% 61 +18 PA 0.50% 54 +31

[0145] Immunoevasive senescent fibroblasts exhibit a significantly higher MICA cleavage rate than senescent fibroblasts. Tested at 0.5% on immunoevasive senescent human fibroblasts, the active ingredient according to the invention (AI) significantly limits MICA cleavage by 31%, thus allowing senescent cells to retain their recognition ligand for the immune system and therefore their elimination.

[0146] Test 3 - Effect of the active ingredient according to the invention on the lysis of immunoevasive senescent fibroblasts.

[0147] The objective of this study is to evaluate the ability of the active ingredient according to the invention (AI) to restore the physiological lysis of immunoevasive senescent fibroblasts by Natural Killer (NK) cells through recognition of the MICA ligand enabling their fixation and lysis of senescent fibroblasts by the action of cytotoxic molecules released by NK cells.

[0148] The protocol is identical to that of trial 1. Once treated with the active ingredient according to the invention, treated or untreated senescent or immunoevasive senescent human fibroblasts are labeled with a whole-cell fluorescent probe and then brought into contact with pre-activated NK cells. Fluorescence is measured. The study thus makes it possible to monitor the morphological evolution of previously labeled senescent or immunoevasive senescent human fibroblasts brought into contact with NK cells.

[0149] The results are presented in Table 7 below.

[0150] [Tables?] NK cell lysis (%) Ability to restore lysis of immunoevasive senescent cells (%) Senescent fibroblasts Control 14.7 Immunoevasive senescent fibroblasts Control 5.1 PA 0.5% 14.8 +101

[0151] Compared to senescent fibroblasts, immunoevasive senescent fibroblasts are significantly less lysed by NK cells. Tested at 0.5% on immunoevasive senescent human fibroblasts, the active ingredient according to the invention significantly restores their lysis by NK cells by 102%, and thus re-establishes the elimination of resistant senescent cells by NK cells.

[0152] Test 4 - Effect of the active ingredient according to the invention on the extracellular matrix.

[0153] The objective of this study is to evaluate the effect of the secretome of immunoevasive senescent fibroblasts pre-treated with the active ingredient according to the invention (AI) on matrix deposition by aged fibroblasts.

[0154] The deposition and structuring of the dermal extracellular matrix (ECM) is one of the main functions of fibroblasts. With age, the structure of the collagen and elastic networks of the dermis is altered, notably due to the accumulation of senescent fibroblasts and the increased secretion of MMP-1 and MMP-3 primarily.

[0155] Aged human fibroblasts are seeded and incubated at 37°C in an atmosphere containing 5% CO2. Then, the aged human fibroblasts are treated either with a senescent fibroblast secretome (SFS), or with an immunoevasive senescent fibroblast secretome (SFSI), treated or untreated with the active ingredient at 0.5% (V / V). The cells are then incubated in an atmosphere containing 5% CO2 at 37°C. Finally, the collagen I network is analyzed by immunocytofluorescence.

[0156] The results are presented in Table 8 below.

[0157] [Tables8] Collagen I network synthesis (xO4 UA) Capacity to preserve collagen I network synthesis (%) Aged fibroblasts SFS Control 1125 SFSI Control 802 SFSI + PA 0.5% 1010 +64

[0158] Collagen I network synthesis by aged human fibroblasts treated with the secretome of immunoevasive senescent fibroblasts is significantly reduced. When applied to aged human fibroblasts, the secretome derived from immunoevasive senescent fibroblasts pretreated with the active ingredient according to the invention at 0.5% significantly preserves collagen I network synthesis by 64%. Thus, by regulating the composition of the SASP secreted by immunoevasive senescent fibroblasts, the matrix environment is preserved.

[0159] Test 5 - Effect of the active ingredient according to the invention on keratin-nocvtary renewal.

[0160] The objective of this study is to evaluate the effect of the secretome of immunoevasive senescent fibroblasts treated with the active ingredient according to the invention (AI) on the renewal of aged keratinocytes.

[0161] With age, epidermal homeostasis is disrupted and the skin struggles to renew itself. Collagen XVII, expressed by basal keratinocytes, plays a crucial role in the proliferation of these cells; its decrease is correlated with skin aging.

[0162] The protocol is identical to that of trial 4 except that the cells are keratinocytes instead of fibroblasts. Collagen XVII synthesis is analyzed by Western blot.

[0163] The results are presented in Table 9 below.

[0164] [Tables9] Collagen XVII Synthesis (UA) Capacity to Preserve Collagen XVII Synthesis (%) Aged Keratinocytes SFS Control 4.1 SFSI Control 2.8 SFSI + PA 0.25% 3.6 +62 SFSI + PA 0.50% 4.0 +92

[0165] Collagen XVII synthesis by aged human keratinocytes treated with the secretome of immunoevasive senescent fibroblasts is significantly reduced. When applied to aged human keratinocytes, the secretome derived from immunoevasive senescent fibroblasts pre-treated with the active ingredient according to the invention at 0.5% significantly preserves collagen XVII synthesis by 95%.

[0166] Thus, by regulating the composition of S ASP secreted by immunoevasive senescent fibroblasts, the active principle according to the invention preserves the renewal of aged human keratinocytes.

[0167] In vivo evaluation of the biological activity of the active ingredient according to the invention

[0168] The objective of the following tests is to evaluate in vivo the biological activities of the active ingredient according to the invention formulated at 2% in emulsion, compared to a placebo formula, on mature Caucasian skin.

[0169] The panel consisted of 18 healthy female volunteers aged 61 to 79 years. The effect was assessed on the face after 28 days of twice-daily application by analyzing dermal density using high-frequency ultrasound, as well as the biomechanical properties of the skin using a Cutometer. Epidermal renewal dynamics were also determined on the forearms of 16 of the volunteers after 28 days of twice-daily application using a dermatoscope.

[0170] Test 6 - Effect of the active ingredient according to the invention on dermal density

[0171] The results of the dermal matrix density after 28 days of biquo application tidienne are presented in table 10 below.

[0172] [TableauxlO] Variation / D0 (%) D28 Placebo -2.8 PA (2%) 8.6 Variation / Placebo (%) 11.4

[0173] Thus, from 28 days of treatment, on mature Caucasian skin, the active ingredient according to the invention formulated at 2% in emulsion significantly improves the density of the dermal matrix by 11.4% compared to placebo.

[0174] Test 7 - Effect of the active ingredient according to the invention on skin elasticity

[0175] The results of skin elasticity after 28 days of twice-daily application are presented in Table 11 below.

[0176] [Tableauxll] Variation / D0 (%) D28 Placebo -0.5 PA (2%) 5.7 Variation / Placebo (%) 6.2

[0177]

[0178] Thus, after just 28 days of treatment, the active ingredient according to the invention, formulated at 2% in emulsion, significantly boosts the elasticity of mature Caucasian skin by 6.2% compared to placebo. This effect was observed in 83% of subjects.

[0179] Test 7 - Effect of the active ingredient according to the invention on epidermal renewal

[0180] The results on epidermal renewal after 28 days of twice-daily application are presented in Table 12 below.

[0181] [Tables l2] Cell renewal (%) Day 2 Day 4 Day 7 Day 9 Day 11 Day 14 Placebo 7 19 38 50 57 70 PA (2%) 9 22 45 57 64 77 Variation / Placebo (%) 22 17 16 15 11 9

[0182] Thus, from 7 days after application of DHA, the active ingredient according to the invention formulated at 2% in emulsion promotes epidermal renewal of Caucasian volunteers by significantly increasing the ability of their skin to eliminate DHA compared to placebo.

[0183] Thus, the active principle according to the invention, formulated at 2% in emulsion, presents rapid and significant effects on the matrix and epidermal dynamics of mature Caucasian volunteers.

[0184] From 28 days of treatment, the active ingredient according to the invention makes it possible to significantly counteract the effects of age on mature skin: - the density of the dermal matrix is ​​increased; - skin elasticity is improved; - Epidermal renewal is accelerated.

[0185] Test 8 - Effect of the active ingredient according to the invention for its anti-wrinkle action

[0186] The results are presented in Table 13 below.

[0187] [Tables 13] Variation / D0 (%) D14 D28 Placebo 0.0 -0.7 PA (2.5%) -7.6 -11.0 Variation / Placebo (%) -7.6 -10.3

[0188] After just 14 days of treatment, the active ingredient according to the invention, formulated at 2% in emulsion, significantly reduces the stage of crow's feet wrinkles in mature Caucasian skin, compared to placebo by 7.6%. This effect is maintained after 28 days of treatment with a significant reduction of 10.3%.

[0189] Test 9 - Effect of the active ingredient according to the invention to enhance the radiance of the complexion

[0190] The results are presented in Table 14 below.

[0191] [Tables 14] Variation / JO (%) J14 J28 Radiation 6.4 6.7 Pink color 12.8 15.7 Olive color -7.7 -12.2 Eye fatigue -5.5 -6.9

[0192] Compared to placebo, the active ingredient according to the invention rapidly and significantly improves the characteristic parameters of complexion radiance in mature Caucasian skin. Indeed, after just 14 days of treatment, the active ingredient according to the invention, formulated at 2% in emulsion, makes the complexion brighter and fresher, and improves the appearance of a healthy glow by reducing oliveness and eye fatigue.

[0193] These effects intensify after 28 days of twice-daily application, with a 6.7% increase in skin radiance and a 15.7% increase in pink color in 89% and 78% of volunteers, respectively. A 12.2% decrease in olive color and a 6.9% reduction in eye fatigue were also observed in 78% of volunteers.

Claims

Demands

1. Cosmetic active ingredient comprising at least one hydrolyzed extract of Ginkgo biloba, characterized in that the extract comprises cyclitols.

2. Cosmetic active ingredient according to the preceding claim, characterized in that the extract is a filtered hydrolysate of Ginkgo biloba.

3. Cosmetic active ingredient according to any one of the preceding claims characterized in that the extract is an enzymatic hydrolysate.

4. Cosmetic active ingredient according to any one of the preceding claims, characterized in that the active ingredient is an extract of Ginkgo biloba leaves.

5. Cosmetic active ingredient according to any one of the preceding claims, characterized in that cyclitols represent at least 5% by weight of the total dry weight of the extract.

6. Cosmetic active ingredient according to any one of the preceding claims, characterized in that the extract comprises carbohydrates.

7. Cosmetic active ingredient according to the preceding claim, characterized in that 20% by weight of the total weight of carbohydrates are carbohydrates having a degree of polymerization between 2 and 34.

8. Cosmetic active ingredient according to any one of the preceding claims, characterized in that it is capable of being obtained by a process comprising the following steps: a. solubilization of at least 50 g / L of Ginkgo biloba in water, and filtration, b. enzymatic hydrolysis, c. separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, d. enzymatic inactivation, and e. optionally concentration and sterilizing filtration.

9. Cosmetic active ingredient according to any one of the preceding claims, characterized in that the active ingredient comprises at least one excipient selected from a preservative, an antioxidant, a stabilizer, an atomizing support, and / or a combination thereof.

10. Cosmetic active ingredient according to any one of the preceding claims for its use in restoring the natural elimination of senescent fibroblasts, or senolysis, by immune cells.

11. Cosmetic use of an active ingredient according to any one of claims 1 to 10, intended for topical application to the skin to reduce wrinkles and / or improve the evenness of the complexion, and / or revive the radiance of the complexion.

12. Cosmetic use according to claim 11 to increase cell renewal and / or dermal density, and / or dermal elasticity.

13. Cosmetic composition for topical application comprising at least one active ingredient according to one of claims 1 to 10 and a physiologically acceptable medium.

14. Cosmetic composition according to the preceding claim, wherein the active ingredient represents at least 0.1% by weight of the total weight of the composition.

15. A process for preparing an active ingredient according to any one of claims 1 to 10 comprising the following steps: a. solubilization of at least 50 g / L of Ginkgo biloba in water, b. enzymatic hydrolysis, c. separation of the soluble and insoluble phases by filtration and recovery of the soluble phase, d. enzymatic inactivation, and e. optionally concentration and sterilizing filtration.