METHOD FOR OBTAINING PLANT EXTRACTS COMPRISING AN AUTOFERMENTATION STEP, COMPOSITIONS COMPRISING SUCH EXTRACTS AND THEIR COSMETIC USES

Abstract

The invention relates to methods for preparing plant extracts comprising an auto-fermentation step and optionally extraction steps enabling the enrichment of the extracts with low molecular weight RNA. The invention further relates to compositions comprising such extracts and their cosmetic uses to combat the signs of skin aging, loss of firmness, improve barrier function and hydration, or brighten the skin. Figure for the abstract: [Fig. 3]

Description

Title of the invention: METHOD FOR OBTAINING PLANT EXTRACTS COMPRISING A SELF-FERMENTATION STAGE, COMPOSITIONS COMPRISING SUCH EXTRACTS AND THEIR COSMETIC USES technical field

[0001] The invention relates to the field of cosmetics and more particularly to the preparation of active extracts of plant origin used in cosmetic formulations to combat the signs of skin aging, loss of firmness, improve barrier function and hydration or even lighten the skin. Technical background of the invention

[0002] The preparation of plant extracts usable in cosmetics uses all the purification techniques known in chemistry and phytochemistry, such as extraction methods using polar or non-polar organic solvents (EP3682865).

[0003] However, there is a growing desire among consumers to turn to natural products containing as few synthetic ingredients as possible. To meet this new demand, the extracts described in this application are 100% natural.

[0004] One approach involves using microorganisms to orchestrate the biotransformation of plants. Numerous processes using selected or modified microorganisms are described in the literature and are used industrially. The most widespread are based on fermentation, known for millennia as a food preservation method. The main advantages of fermentation are preservation, improved taste, and enhanced nutritional quality of food, due to the presence of a higher quantity of easily assimilated bioactive molecules.

[0005] In the field of cosmetics, microorganisms are widely used for their direct beneficial effects on the skin or to produce fermented or biotransformed plant extracts. Examples include documents EP3744339A1 and CN110680774A, which describe plant fermentation processes, specifically for Citrus aurantium and jasmine flowers respectively, involving inoculation with yeasts of the species Saccharomyces cerevisiae, or the document CN101243897A which describes the use of lactic acid bacteria to produce cosmetic or food flower extracts.

[0006] In the same field, document FR3103826 describes the preparation of a consortium of microorganisms composed of at least one lactic acid bacterium, one yeast and one acetic acid bacterium, to prepare plant extracts rich in active compounds, suitable for use in the pharmaceutical, cosmetic and food fields.

[0007] Biotechnological processes using microorganisms on synthetic nutrient media adapted to produce compounds of interest such as vitamins, citric acid, lactic acid or vanillin (US 20060269632) are also described.

[0008] However, the processes mentioned above involve the exogenous supply of microorganisms or the supply of specific nutrients to stimulate the growth of microorganisms which will ensure the biotransformation of plant matter.

[0009] Fermentation can also rely on the endogenous microflora present on the surface or in the internal structures of plants. This is a community of mutualistic or symbiotic bacterial and fungal microorganisms called phytobiota, hosted by virtually all plants or parts of plants (fruits, leaves, flowers, stems, roots, seeds, fungi, algae). This is then referred to as spontaneous fermentation or autofermentation.

[0010] Thus, document US20060269632A1 describes the preparation of an effective allergy drug obtained by mixing pine shoots with water and sugars and allowing spontaneous fermentation to develop for several months under anaerobic conditions.

[0011] One problem that the invention proposes to solve is to provide a new process for preparing natural plant extracts, simple to implement, without the addition of any living microorganism, any enzyme, or any exogenous nutrient, and yet exhibiting remarkable biological efficiency and an absence of toxicity.

[0012] To this end, the inventors have developed a process comprising a controlled self-fermentation step carried out essentially from the phytobiota of plant materials, including flowers, fruits, leaves, roots and fungi, without any exogenous input.

[0013] Under these conditions, a significant amount of organic acids and phenolic acids are generated. The resulting plant extracts then contain a wide range of phytomolecules, giving them proven biological efficacy.

[0014] The extracts thus obtained can be used in cosmetics for skin care and more particularly to combat the signs of skin aging, loss of firmness or tone, improve the barrier function and hydration, and brighten the complexion. skin or even improve the skin's innate immune defenses and in vivo well-being. Summary of the invention

[0015] The invention relates to a process for obtaining plant extracts enriched in organic acids and phenolic acids, comprising the following steps:

[0016] a) the plant material is brought into contact with water,

[0017] b) the pH is adjusted if necessary to a value between 4 and 9,

[0018] c) the mixture is kept under gentle stirring for a time between 6 hours and 48 hours, at a temperature between 20 and 60°C, at a pH between 4 and 7, to allow the self-fermentation process, in an enclosure allowing gas exchange with the atmosphere.

[0019] Optionally continued by the following steps:

[0020] d) the mixture obtained in c) is purified to remove residual solid plant matter and collect the liquid portion,

[0021] e) at least one filtration of the liquid portion obtained in the previous step is carried out,

[0022] f) the pH of the filtrate is checked and readjusted, if necessary, to a value between 4 and 8,

[0023] g) the filtrate is sterilized,

[0024] h) the extract is diluted with a physiologically acceptable solvent.

[0025] The invention further relates to a process for preparing extracts enriched in organic acids and phenolic compounds, obtained from jasmine flowers of the species Jasminum grandiflorum or violet flowers of the species Viola odorata, comprising the following preceding steps a) and c) followed by the following steps:

[0026] To the mixture obtained in step c) phytic acid is added at a concentration between 1 and 5 mM, at a pH between 10 and 11, the pH of the mixture obtained is adjusted to a value between 6 and 8, the residual plant matter is then separated, the liquid fraction obtained is purified by successive filtrations to clarify the extract and optionally, the extract is diluted with a physiologically acceptable solvent.

[0027] The invention further relates to diluted plant extracts obtained by one of the preceding processes, having 2 to 40 g / kg of dry weight; 0.2 to 30 g / kg of sugars; 10 to 4000 mg / kg of organic acids; 10 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / L of amino acids.

[0028] The invention further relates to a composition comprising an effective quantity of extract obtained by one of the preceding processes and a physiological medium.

[0029] The invention also relates to the cosmetic use of the previous composition to combat the signs of skin aging, loss of firmness, and improve the barrier function and hydration, brightening the skin, mitigating the age-related decline in the number or activity of mechanosensory touch receptors (Piezol) and oxytocin receptors (OXTR), or improving the skin's innate immune defenses and in vivo well-being. Brief description of the drawings

[0030] [Fig. 1] Protein profiles of jasmine extracts by HPLC

[0031] [Fig.2] Evolution of phenolic compounds in jasmine extract by HPLC

[0032] [Fig. 3] Analysis of shikimic acid in jasmine extract by HPLC

[0033] [Fig.4] Analysis of the total sugar content of the violet extract

[0034] [Fig. 5] Analysis of organic acids in violet extract by HPLC

[0035] [Fig.6] Evaluation of jasmine extract on the expression of the piezolytic receptor in reconstructed epidermis

[0036] [Fig.7] Evaluation of jasmine extract on the expression of the oxytocin receptor (OXTR) on human skin biopsies

[0037] [Fig.8] Evaluation of jasmine extract on the expression of oxytocin receptor (OXTR) messenger RNA in cultured keratinocytes

[0038] [Fig.9] Evaluation of jasmine extract on viperin expression in cultured keratinocytes

[0039] [Fig. 10] Evaluation of jasmine extract on the level of E-cadherin expression, on human skin biopsies pretreated with a piezolytic activity blocker (Dookul)

[0040] [Fig. 11] Evaluation of jasmine extract on the expression level of messenger RNA of the enzyme 11[3-HSD1] in cultured keratinocytes. Detailed description of the invention. Definitions.

[0041] All terms used in this description have their most widely understood meanings unless otherwise stated. For the purposes of the invention, the following terms are defined as follows:

[0042] The term “extract” means the result of all aqueous extraction processes of plant material.

[0043] By "phytobiota" we mean all the microorganisms present on the surface or inside plant matter.

[0044] In the present description, "plant matter" or "plant" means a living organism belonging to the plant kingdom and possessing a phytobiota, including, among others, plants, fungi, mosses, lichens and algae.

[0045] The term "fresh plant material" means that the plant material used in the extraction process has not undergone any chemical or mechanical treatment likely to alter its phytobiota. For example, the plant material was harvested shortly before being used in the process, or it was frozen quickly after harvesting, or it was dried under conditions that allow for good preservation of the phytobiota, for example at low temperature. Fresh plant material may include plant residues obtained after processing.

[0046] "Auto-fermentation" means a fermentation process resulting from the plant material naturally present in or on the surface of the plant material involved in the extraction process described in this application.

[0047] By "proteins" we mean large molecules composed of chains of amino acids, but also polypeptides and peptides smaller than 20 kDa (or 20 kg / mol).

[0048] By "small RNAs" or "small molecular weight RNAs" is meant a mixture of non-coding RNAs (ribonucleic acids), of small molecular weight, of a length of no more than 150 nucleotides, such as all types of small non-messenger RNAs, single and / or double stranded, for example microRNAs, interfering RNAs, introns, small nuclear RNAs or any RNA fragment.

[0049] By "organic acids" is meant derivatives of the catabolism of amino acids, fatty acids and sugars comprising an acid function. This group includes α-hydroxylated carboxylic acids (AHAs) or polyhydroxylated carboxylic acids such as glycolic, lactic, malic, citric, tartaric, shikimic acids, carboxylic acids derived from fruit sugars or from any other parts of plants such as uronic acids, or diacids such as succinic acid.

[0050] By "phenolic compounds" we mean all molecules possessing one or more aromatic rings themselves bearing one or more hydroxyl groups, such as phenolic acids, flavonoids or their derivatives, tannins or any other polyphenols.

[0051] By "phenolic acids" we mean phenolic compounds derived from benzoic acid and cinnamic acid, molecules possessing a single aromatic ring.

[0052] By "flavonoids" we mean phenolic compounds all sharing the same basic structure formed by two aromatic rings linked by three carbons.

[0053] By "glycosylated flavonoids" we mean flavonoids linked to one or more sugars.

[0054] By "sugars" we mean all carbohydrates such as mono- and disaccharides as well as oligo- and polysaccharides contained in the extract.

[0055] By "phytomolecules of interest", we mean all the molecules present in the extracts of the invention and in particular, proteins, sugars, phenolic compounds, organic acids, amino acids, small RNAs of a maximum length of 150 nucleotides.

[0056] When a range of values ​​is described, the bounds of that range must be understood as explicitly including all intermediate values ​​in the range. For example, a range of values ​​between 1% and 10% must be understood as including 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, as well as all decimal values ​​between 1% and 10%.

[0057] The numerical values ​​in percentage are percentages by weight, that is to say the weight of a compound in relation to the total weight of the mixture envisaged, unless otherwise specified.

[0058] The compositions described in this application may “include”, “consist of” or “consist essentially of” the essential compounds or optional ingredients.

[0059] “Consist essentially of” means that the composition or component may include additional ingredients, but only if the additional ingredients do not alter the basic characteristics or new characteristics of the composition or use described in this application.

[0060] A "physiologically acceptable medium" means a vehicle suitable for contact with the outer layers of the skin or mucous membranes, without causing toxicity, irritation, undue allergic response and similar or intolerance reaction, and proportionate to a reasonable benefit / risk ratio.

[0061] “Skin” means the skin of the face and body including the scalp (including hair follicles and inter-follicular skin spaces) and skin appendages (hair, body hair and nails).

[0062] By "effective quantity" is meant the minimum quantity of extract according to the invention which is necessary to obtain at least one of the desired biological activities, without this quantity being toxic.

[0063] By "skin hydration" is meant the content and distribution of water in the upper layers of the epidermis.

[0064] By "signs of skin aging" we mean all changes in the external appearance of the skin due to aging such as, for example, wrinkles and fine lines, creases, bags under the eyes, dark circles, sagging, loss of elasticity, firmness and / or tone of the skin, pigmentary disorders such as senile lentigo or solar lentigo, but also all internal changes in the skin which do not systematically result in a changed external appearance such as, for example, the decrease in the number or activity of mechanosensory touch receptors (Piezol) and oxytocin receptors (OXTR), thinning of the skin, or all internal degradation of the skin resulting from environmental stresses such as pollution and solar radiation including UV.

[0065] By "in vivo well-being" we mean the emotional state of a person, the subjective feeling of well-being, the physiological production of oxytocin and the measurement of behavior or emotional expression.

[0066] It is understood that the invention relates to mammals and more particularly to human beings. Autofermentation and extraction processes

[0067] The invention primarily relates to a process for obtaining plant extracts enriched in organic acids and phenolic acids, comprising the following steps:

[0068] a) the plant material is brought into contact with water,

[0069] b) the pH is adjusted if necessary to a value between 4 and 9,

[0070] c) the mixture is kept under gentle stirring for a time between 6 hours and 48 hours, at a temperature between 20 and 60°C, at a pH between 4 and 7, to allow the self-fermentation process, in an enclosure allowing gas exchange with the atmosphere.

[0071] It is the fresh plant material placed in the presence of water that provides the nutrients necessary for the growth of the phytobiota. The nutrient supply is sufficient to allow the development and survival of the phytobiota for several days.

[0072] The process of the invention is implemented without the addition of exogenous nutrients and does not use detergents or solvents that are potentially toxic in cosmetics. It therefore has a reduced impact on the environment.

[0073] Moderate agitation helps to keep the microorganisms of the phytobiota in suspension in the mixture.

[0074] The self-fermentation process according to the invention is carried out in an open environment to allow gas exchange with the atmosphere, without any particular constraint to ensure sterility, but can alternatively be implemented in a fermenter in order to control gas exchange.

[0075] Auto-fermentation takes place when the microorganisms of the phytobiota, through their enzymes, metabolize nutrients of plant origin, such as sugars, proteins, polyphenols to produce an extract rich in various families of simpler and more easily assimilated compounds such as amino acids, organic acids, peptides and phenolic acids.

[0076] Analytical tests, carried out in parallel with a study of the kinetics of microbial growth, made it possible to observe a significant modification of the environment during self-fermentation such as a decrease in the concentration of flavonoids, a decrease in the concentration of sugars, and the appearance of fermentation markers such as phenolic and organic acids attesting to the fermentative activity of the microorganisms.

[0077] In a particular embodiment, the plant materials used are plants from the Oleaceae, Poaceae, Fabaceae families or fungi from the Tuberaceae family.

[0078] In another particular embodiment, the plant materials used are plants of the Oleaceae family such as jasmine of the species Jasminum grandiflorum, and of the Violaceae family such as the species Viola odorata.

[0079] In a particular embodiment, the plant parts used are chosen from among the flowers, fruits, leaves and roots.

[0080] In a preferred embodiment, the plant parts used are the flowers.

[0081] In another embodiment, the plant materials used are Fungi of the Tuberaceae family and preferably the plant material used are chosen from the species Tuber melanosporum and Tuber magnatum. In this case, the parts of the fungi used are chosen from the mycelium and the fruiting bodies, and more preferably the fruiting bodies.

[0082] In another embodiment, the plant materials used are algae.

[0083] In another particular embodiment, the plant material is a residue of plant obtained after processing such as oilseed cakes or spent grains.

[0084] In step a) distilled, demineralized or mineral-rich and / or trace-element water is used. Distilled water is preferred.

[0085] The plant material used in step a) may be whole or mechanically ground using a blade mill, for example, to reduce the particle size from 0.5 mm to a few centimeters without affecting the viability of the microorganisms. Grinding may be carried out dry or in water.

[0086] When the plant material is composed of flowers, it is preferably used whole, that is to say, uncrushed. Indeed, comparative extractions carried out with and without crushing have shown that the passive diffusion of the compounds contained in the flowers is sufficiently high to make crushing unnecessary.

[0087] On the other hand, when the starting plant material is too thick or composed of large fragments, for example the fruiting part of a mushroom, a whole alga or a root, it is preferably ground beforehand before step a).

[0088] At step a) the ratio of plant matter to water is preferably between 3 and 30% by weight / weight, more preferably between 3 and 15%, even more preferably it is 3%, 5% or 15%.

[0089] At the beginning of step b) the pH is checked and adjusted to a value between 4 and 9, preferably between 6 and 8 and even more preferably to pH 7, by the addition of hydrochloric acid (HCl), citric acid or sodium hydroxide (NaOH). The pH is then checked regularly throughout step b) and readjusted if necessary.

[0090] When citric acid is used at this stage, this quantity of citric acid is removed in the final calculations of organic acid concentration of the extract.

[0091] In step c), the mixture is advantageously stirred moderately. The temperature is adjusted and maintained between 20 and 60°C, more preferably between 20 and 40°C, and even more preferably between 30 and 40°C. These temperature ranges are ideal for ensuring optimal microorganism development; indeed, below 20°C microbial growth is slowed and above 60°C the viability of microorganisms decreases.

[0092] Step c) must be carried out at an optimal pH and temperature to enrich the aqueous extraction solution with phytomolecules and create the growth conditions for the phytobiota present in the mixture. During this step, the pH must not fall below 5.

[0093] Step c) of auto-fermentation takes place over a period of between 1 and 48 hours, preferably between 6 and 30 hours, and more preferably between 15 and 30 hours. A study of microbial growth kinetics demonstrated that growth is strong, under the process conditions, for a period of at least 30 hours, then reaches a plateau.

[0094] Optionally, at the end of step c) a step c') of grinding the residual solid matter can be carried out, in order to release the cellular contents of the microorganisms and thus increase the yield of phytomolecule extraction.

[0095] Optionally, the process can be continued with the following steps:

[0096] d) the mixture obtained in c) is purified to remove residual solid plant matter and collect the liquid portion,

[0097] e) at least one filtration of the liquid portion obtained in the previous step is carried out,

[0098] f) the pH of the filtrate is checked and readjusted, if necessary, to a value between 4 and 8, and preferably to a value between 4 and 7.

[0099] g) The filtrate is sterilized.

[0100] h) The crude extract thus obtained can be diluted in a solvent physiologically acceptable for cosmetic use.

[0101] Step d) of purifying the mixture obtained at the end of step c) removes the residual solid matter and collects the liquid portion. Any method known to those skilled in the art may be used. The mixture obtained in c) may, for example, be filtered through filters with a porosity greater than 30 µm. Preferably, the mixture obtained in c) is centrifuged at low speed, for example for at least 10 min at 4000 g.

[0102] In step e), the liquid portion obtained in step d) is preferably subjected to successive filtrations by lowering the filtration threshold from 30 to 1 µm in order to clarify the solution. Preferably, at least 4 filtrations are carried out with filters of porosity ranging from 30pm, 4pm, 2 and Ipm.

[0103] In step f), the pH of the mixture obtained in the previous step is adjusted to a value between 4 and 8, and preferably between 4 and 7. The pH can be adjusted by adding a solution of sodium hydroxide (NaOH) or hydrochloric acid (HCl), or any other equivalent acid compatible with cosmetic use, such as citric acid. This step is essential to prevent the precipitation of phytomolecules of interest, such as sugars, phenolic compounds, organic acids, proteins, and amino acids, and thus to obtain a stable extract.

[0104] At step g), the filtrate can be sterilized by any method known to those skilled in the art, for example by steaming or preferably by sterilizing filtration through a 0.2 to 0.45 pM filter.

[0105] At the end of step g) a concentrated crude extract is obtained.

[0106] The auto-fermentation process allows the enrichment of the extract in phenolic acids from the transformation of polyphenols and in organic acids such as lactic, succinic and shikhnic acid.

[0107] Preferably, the concentrated crude extracts obtained by the process described above contain at least 300 mg / kg of phenolic compounds and at least 300 mg / kg of organic acids.

[0108] Using the process of the invention according to steps a) to g), concentrated crude extracts are obtained having a dry weight of 4 to 60 g / kg, containing 0.5 to 40 g / kg of sugars, 50 to 5000 g / kg of organic acids, 50 to 5000 mg / kg of phenolic compounds, and 30 to 3000 mg / L of amino acids. However, having undergone various treatments, the extracts obtained may exhibit significant variability depending on factors such as the location or year of harvest, the season, climatic conditions, biotic stress, etc.

[0109] In step h), among the physiologically acceptable solvents, one can mention water, glycerol, ethanol, propanediol and its natural version derived from maize, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycols, cyclic polyols, or any mixture of these solvents. One of the advantages of such dilution, besides obtaining precisely the desired phytomolecule concentrations, is to improve the stability and preservation of the self-fermented plant extract.

[0110] Preferably, the concentrated crude extract is diluted with butylene glycol, propanediol, or glycerin, and even more preferably diluted by the addition of 30% propanediol to ensure its stability and preservation over time by preventing contamination. The crude extract can also be diluted by the addition of 30% glycerin combined with any type of water-soluble preservative such as sodium benzoate or potassium sorbate at a final concentration of 0.5%, or phenoxyethanol at a final concentration of 1.5%.

[0111] This so-called diluted extract comprises, by weight of the total weight of the extract, 2 to 40 g / kg of dry extract, 0.2 to 30 g / kg of sugars, 10 to 4000 mg / kg of organic acids, 10 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / L of amino acids.

[0112] In addition, between step c) or c') and step g), further extraction steps may be carried out according to any process known to the person skilled in the art.

[0113] Among these additional extraction steps, we can mention an extraction process allowing enrichment in small molecular weight RNA described in patents FR 1670672 and US 11,021,505.

[0114] To carry out this process, the self-fermented plant material obtained in step c), i.e. comprising the liquid fraction and the ground plant residues, is brought together with phytic acid at a concentration between 1 and 5 mM, at a pH between 10 and 11, the pH of the resulting mixture is adjusted to a value between 6 and 8. The residual plant material is separated and the liquid fraction obtained is purified by successive filtrations to clarify the extract.

[0115] The extract can then be sterilized and diluted according to steps g) and h) identical to those of the process described above.

[0116] In this particular embodiment, the plant material is advantageously a flower such as the jasmine flower of the species Jasminum grandiflorum or the violet of the species Viola odorata.

[0117] One of the advantages of combining the autofermentation process with a complementary extraction process that allows for enrichment in small RNAs is to increase the extraction yield by a factor of 2 or 3 compared to an extract obtained by autofermentation. The concentration of proteins, sugars, organic acids, and total phenolic compounds is also increased by at least a factor of 2.

[0118] Thus the concentrated crude extracts obtained by the process described above have a dry weight at least 2 times greater than the dry weight of the extracts obtained by simple autofermentation and contain at least 400 mg / kg of phenolic compounds and at least 500 mg / kg of organic acids.

[0119] Among these additional extraction steps, we can also mention one of the many extraction processes known to those skilled in the art involving one or more successive enzymatic hydrolyses. Plant extracts obtained

[0120] The invention has as its second object a plant extract obtained by the auto-fermentation process comprising steps a) to g) and comprising at least 300 mg / kg of phenolic compounds.

[0121] The invention also relates to a concentrated plant extract, characterized in that it comprises, by weight of the total weight of the extract, 4 to 20 g / kg of dry extract, 0.5 to 40 g / kg of sugars, 50 to 4000 mg / kg of organic acids, 50 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / kg of amino acids, which can be obtained by the process described above.

[0122] The invention further relates to a plant extract, characterized in that it comprises, by weight of the total weight of the extract, 4 to 20 g / kg of dry extract, 0.5 to 40 g / kg of sugars, 50 to 4000 mg / kg of organic acids, 50 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / kg of amino acids, obtained by the process described above.

[0123] The extract of the invention thus comprises a wide range of phytomolecules that may have beneficial effects on the skin, without presenting a risk of skin irritation or other harm to health.

[0124] In a particular embodiment, the extract of the invention contains at least 300 mg / kg of organic acids.

[0125] In a particular embodiment, the extract of the invention contains at least 100 mg / kg of specific fermentation acids, selected from lactic acid and shikimic acid.

[0126] In particular, the extract of the invention contains at least 300 mg / kg of phenolic compounds.

[0127] In a particular embodiment, the invention relates to a diluted extract comprising, by weight of the total weight of the extract, 2 to 40 g / kg of dry extract, 0.2 to 30 g / kg of sugars, 10 to 4000 mg / kg of organic acids, 10 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / L of amino acids.

[0128] The extract of the invention does not contain ethanol since the process of the invention is not an alcoholic type fermentation.

[0129] In a particular embodiment, the invention relates to a plant extract obtained by the self-fermentation process comprising steps a) to c) coupled with additional extraction steps to allow enrichment in small molecular weight RNA, but also to increase the extraction yield by a factor of 2 or 3.

[0130] Advantageously in this particular embodiment the extract is obtained from fresh jasmine flowers (Jasminum grandiflorum) or fresh violet flowers (Viola odorata), and has a dry weight at least 2 times greater than the dry weight of extracts obtained by simple autofermentation and comprises at least 400 mg / kg of phenolic compounds and at least 500 mg / kg of organic acids. cosmetic compositions

[0131] A third object of the invention is a cosmetic composition comprising an effective amount of at least one plant extract obtained according to the process described in this application.

[0132] Advantageously, the plant extracts of the invention are used in diluted form and are added to a physiologically acceptable medium at a concentration of 0.05 to 5% by weight relative to the total weight of the composition, preferably at a concentration of 0.1 to 2.5% by weight relative to the total weight of the composition.

[0133] The composition usable according to the invention is formulated to be applied by any suitable route, in particular oral, or external topical, and the formulation of the compositions will be adapted by a person skilled in the art.

[0134] Preferably, the compositions according to the invention are in a form suitable for topical application. These compositions must therefore contain a physiologically acceptable medium, i.e., one compatible with the skin and its appendages, without risk of discomfort during application, and cover all suitable cosmetic forms.

[0135] The compositions for implementing the invention may in particular be in the form of an aqueous, hydroalcoholic or oily solution, an oil-in-water emulsion, a water-in-oil emulsion or multiple emulsions; they may also be in the form of suspensions, or even powders, suitable for application on the skin, mucous membranes, lips and / or hair.

[0136] These compositions may be more or less fluid and may also have the appearance of a cream, lotion, milk, serum, ointment, gel, paste or mousse. They may also be in solid form, such as a stick, or be applied to the skin as an aerosol.

[0137] Examples of physiologically acceptable media commonly used in the intended field of application include formulation aids such as solvents, thickeners, diluents, antioxidants, colorants, sunscreens, self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0138] In all cases, a person skilled in the art shall ensure that these additives and their proportions are chosen in such a way as not to impair the desired advantageous properties of the composition according to the invention. These additives may, for example, represent 0.01 to 20% of the total weight of the composition. When the composition according to the invention is an emulsion, the oil phase may represent from 5 to 80% by weight, and preferably from 5 to 50% by weight, relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the composition are chosen from those commonly used in the relevant field. For example, they may be used in a proportion ranging from 0.3 to 30% by weight relative to the total weight of the composition.

[0139] According to another advantageous embodiment of the invention, the plant extracts of the invention can be encapsulated or included in a cosmetic vector such as liposomes or any other nano capsule or microcapsule used in the field of cosmetics or adsorbed on powdered organic polymers, mineral supports such as talcs and bentonites.

[0140] Advantageously, the composition according to the invention may include, in addition to the plant extract according to the invention, at least one other active agent having cosmetic effects similar and / or complementary to those of the invention.

[0141] For example, the additional active agent(s) may be chosen from: anti-aging, firming, brightening, moisturizing, draining, microcirculation-enhancing, exfoliating, desquamating, extracellular matrix-stimulating, energy-metabolizing, antibacterial, antifungal, soothing, free radical-scavenging, anti-UV, anti-acne, anti-inflammatory, anesthetic, warming, cooling, slimming agents.

[0142] Such additional active agents may be selected from groups including:

[0143] - vitamins (vitamin A and its derivatives; vitamins B3, B5, B6 and B12; vitamin C; vitamins E, F, H, K, PP or even coenzyme Q10);

[0144] - metalloproteinase inhibitors, or TIMP activators;

[0145] - DHEA, its precursors and derivatives;

[0146] - amino acids, natural or synthetic peptides,

[0147] - Artemia salina extract, marketed under the name GP4G™ (FR2817748, ASHLAND®);

[0148] - plant peptide extracts, yeast extracts, extracts of polyphenols;

[0149] - dehydroacetic acid (DHA);

[0150] - phytosterols of synthetic or natural origin;

[0151] - salicylic acid and its derivatives, alpha- and beta-hydroxy acids, silanols;

[0152] - amino sugars and polysaccharides;

[0153] - lipids such as ceramides or phospholipids;

[0154] - cyclic AMP and its derivatives, the methyl xanthines. cosmetic uses

[0155] The invention has as its fourth object the cosmetic use of a composition comprising the plant extracts of the invention for skin care, more particularly to combat the signs of skin aging, loss of firmness or tone, improve barrier function and hydration, lighten the skin, mitigate the age-related decrease in the number or activity of mechanosensory touch receptors (Piezol) and oxytocin receptors (OXTR), or improve the skin's innate immune defenses and in vivo well-being.

[0156] Preferably, the cosmetic uses according to the present invention relate to cosmetic treatment methods by topical applications on healthy skin.

[0157] In particular, the invention relates to the use of self-fermented extracts of fresh jasmine flowers (Jasminum grandiflorum) or fresh violet flowers (Viola odorata).

[0158] The plant extracts of the invention have been tested on biological markers associated with aging and hydration, such as the expression of collagen and hyaluronic acid, and have demonstrated superior efficacy to conventional control plant extracts.

[0159] The plant extracts of the invention have been tested on biological markers associated with skin firmness or tone, such as e-cadherin, and have demonstrated superior efficacy to conventional control plant extracts.

[0160] The plant extracts of the invention have been tested on tyrosinase, an enzyme associated with melanin synthesis, and therefore with skin pigmentation, and have demonstrated superior tyrosinase inhibitory activity compared to conventional control plant extracts.

[0161] In a particular embodiment, the invention relates to the cosmetic use of a composition comprising a self-fermented extract of fresh jasmine flowers (Jasminum grandiflorum) obtained by a self-fermentation process coupled with an extraction process allowing specific enrichment in small molecular weight RNA (process described in patents FR 1670672 and US 11,021,505).

[0162] In this particular embodiment, the self-fermented extract then extracted from fresh jasmine flowers (Jasminum grandiflorum) is used to mitigate the age-related decline in the number or activity of mechanosensory touch receptors (Piezol) and oxytocin receptors (OXTR), improve the skin's innate immune defenses and in vivo well-being.

[0163] In this particular embodiment, jasmine flower extracts have demonstrated their ability to positively modulate touch receptors present in the skin (Piezol).

[0164] In this particular embodiment, jasmine flower extracts have demonstrated their ability to stimulate the expression of the oxytocin receptor, a molecule involved in preventing skin aging.

[0165] In this particular embodiment, jasmine flower extracts have demonstrated their ability to stimulate the expression of viperine.

[0166] Indeed, keratinocytes are the body's first line of defense, initiating an innate immune response by recognizing pathogens. The extract The jasmine of the invention was tested on the expression of viperin, which is one of those factors of innate immune defense synthesized by keratinocytes.

[0167] In this particular embodiment, a composition comprising 2% jasmine flower extracts, applied topically by healthy volunteers, produced a positive effect on well-being in vivo. In this test, emotional state was assessed through three distinct components: the subjective feeling of well-being, salivary oxytocin production, and the measurement of emotional behavior or expression (Don Hockenbury and Sandra E. Hockenbury, Discovering Psychology, 5th edition, Page 344, Chapter 8, Worth Publishers). Examples

[0168] By way of illustration, examples of embodiments of the process according to the invention are described below.

[0169] Example 1: Preparation of an extract from self-fermented fresh jasmine (Jasminum grandiflorum) flowers

[0170] The species Jasminum grandiflorum belongs to the genus Jasminum and is widely used in industry as a fragrance or aroma.

[0171] In a first step a), 200 g of whole fresh jasmine flowers are mixed with 1800 g of distilled water, i.e. 10% of raw material involved in the process and 90% water for a total weight of 2 kg.

[0172] b) The pH is adjusted to 7.

[0173] c) The mixture is maintained at 30°C in an open beaker to maintain an aerobic atmosphere, favorable to the development of microorganisms for 24 hours. During this step, the pH is monitored regularly, showing an acidification of the medium linked to the development of microorganisms, without it falling below 5.

[0174] d) The mixture obtained is centrifuged for 10 min at 4000 g, so as to sediment the residual plant matter in the pellet and collect the supernatant.

[0175] e) The resulting mixture is then subjected to successive filtrations by lowering the filtration threshold from 30 to 0.2 pm.

[0176] f) The pH of the filtrate obtained is adjusted to 6.3 using 10% concentrated citric acid.

[0177] g) The filtrate is sterilized by filtration.

[0178] The undiluted extract has a dry weight of 11.7 g / kg and contains a concentration of 5.2 g / kg of total sugars, 3.7 g / kg of protein, 411.1 mg / kg of total organic acids, 630 mg / kg of total phenolic compounds.

[0179] h) The extract is diluted with plant-derived glycerin to obtain a final concentration of 30% glycerin and 70% self-fermented jasmine flower extract.

[0180] The diluted extract has a dry weight of 8.1 g / kg, and exhibits a concentration of 3.6 g / kg of total sugars, 2.6 g / kg of proteins, 287.7 mg / kg of total organic acids, 441 mg / kg of total phenolic compounds.

[0181] Example 2: Preparation of an extract of self-fermented fresh jasmine flowers (Jasminum grandiflorum) by self-fermentation coupled with a second extraction process allowing specific enrichment in small RNAs

[0182] Once the self-fermentation has been carried out according to steps a) to c) of example 1, 2g / L or 3 mM of phytic acid is added to this self-fermented medium.

[0183] The pH is adjusted to pH 11 to allow enrichment of the extract in small molecular weight RNA as well as in various phytomolecules.

[0184] The mixture is heated for 2 hours at 80°C under stirring.

[0185] The mixture is then filtered using 30 µm porosity filters to separate the solid matter of the filtrate.

[0186] Sequential filtrations on filters of decreasing porosity are then carried out in order to clarify the self-fermented plant extract up to a filtration at 1 pm of porosity.

[0187] The pH is adjusted to 6.3 with a citric acid solution.

[0188] At this stage, the extract has a dry weight of 17.5 g / kg, and exhibits a concentration of 6.4 g / kg of total sugars, 6.6 g / kg of protein, 651.8 mg / kg of total organic acids, 1301 mg / kg of total phenolic compounds and 45 mg / kg of low molecular weight RNA and is devoid of DNA.

[0189] The absence of DNA was demonstrated by a DNase test, an enzyme that specifically degrades DNA and not RNA. The electrophoretic profile after DNase action is not altered, demonstrating that the nucleic acid present in the extract is not sensitive to DNase and is therefore not DNA.

[0190] The extract is then diluted with plant-derived glycerin to obtain a final concentration of 30% glycerin and 70% self-fermented jasmine flower extract.

[0191] The diluted extract has a dry weight of 11.7 g / kg, and exhibits a concentration of 4.1 g / kg of total sugars, 4.6 g / kg of protein, 413.7 mg / kg of total organic acids, 874 mg / kg of total phenolic compounds and 31 mg / kg of small molecular weight RNA.

[0192] Example 3: Preparation of an extract of self-fermented fresh violet flowers (viola odorata)

[0193] Plants of the genus Viola are herbaceous plants belonging to the family Violaceae. The species Viola odorata is known to be the only one of the genus Viola that is fragrant and is therefore widely used in the perfume industry, in confectionery, and in cooking. This species also has recognized medicinal properties in herbalism.

[0194] In a first step a), 200 g of whole and fresh violet flowers are mixed with 1800 g of distilled water, i.e. 10% of raw material involved in the process and 90% water for a total weight of 2 kg.

[0195] Steps b) to g) are strictly identical to example 1.

[0196] The extract obtained has a dry weight of 7.1 g / kg, and a concentration of 1.0 g / kg of total sugars, 1278.0 mg / kg of total organic acids, 444.5 mg / kg of total phenolic compounds and 2.1 g / kg of protein.

[0197] The extract is then diluted with plant-derived glycerin to obtain a final concentration of 30% glycerin and 70% self-fermented violet flower extract.

[0198] The extract thus diluted has a dry weight of 5.0 g / kg, and exhibits a concentration of 0.7 g / kg of total sugars, 894.6 mg / kg of total organic acids, 344.4 mg / kg of total phenolic compounds and 1.5 g / kg of proteins.

[0199] Example 4: Preparation of an extract of fresh violet flowers (Viola odorata) by autofermentation coupled with a second extraction process allowing specific enrichment in small RNAs

[0200] Once the auto-fermentation has been carried out according to steps a) to c) of example 3, 2g / L or 3 mM of phytic acid is added to this phytofermented medium.

[0201] The pH is adjusted to pH 11 to allow enrichment of the extract in small molecular weight RNA as well as in various phytomolecules.

[0202] The mixture is heated for 2 hours at 80°C under stirring.

[0203] Sequential filtrations on filters of decreasing porosity are then carried out in order to clarify the self-fermented plant extract up to a filtration at 1 pm of porosity.

[0204] The pH is adjusted to 6.3 with a citric acid solution.

[0205] At this stage, the extract has a dry weight of 14.8 g / kg, and a concentration of 2.45 g / kg of total sugars, 1336.9 mg / kg of total organic acids, 492.2 mg / kg of total phenolic compounds, 3.95 g / kg of protein and 26 mg / kg of small molecular weight RNA and does not contain DNA.

[0206] The extract is then diluted with plant-derived propanediol to obtain a final concentration of 30% propanediol and 70% self-fermented violet flower extract.

[0207] The extract thus diluted has a dry weight of 10.36 g / kg, and a concentration of 1.7 g / kg of total sugars, 935.8 mg / kg of total organic acids, 344.4 mg / kg of total phenolic compounds, 2.8 g / Kg of protein and 18 mg / kg of small molecular weight RNA and does not contain DNA.

[0208] Example 5: Preparation of conventional extracts of fresh flowers as comparators

[0209] For comparative purposes, conventional extracts were made using the same quantity of whole, fresh flowers as in examples 1 to 4, i.e. 10% of plant raw material in distilled water.

[0210] The mixture is then heated for 1 hour at 25°C, and then filtered by first filtration through large porosity filters of 30 µm to remove residual solid plant matter from the liquid portion. Sequential filtrations through filters of decreasing porosity are then carried out to clarify the plant extract until a sterile filtration at 0.2 µm porosity is achieved.

[0211] The liquid portion obtained in the previous step constitutes a conventional control extract.

[0212] Comparative analytical data could thus be generated, as illustrated in figures [Fig.1], [Fig.2], [Fig.3], [Fig.4], [Fig.5] and examples 6, 7, 9 and 10.

[0213] Example 6: Analysis of jasmine flower extracts from Examples 1 and 2 versus a conventional extract

[0214] The analyses were carried out on the extracts of examples 1 and 2 before dilution and on a conventional extract of example 5.

[0215] Methodologies

[0216] The total phenolic content of the extracts was measured by spectrophotometry at 760 nm after reduction of the Folin-Ciocalteu reagent by the phenols. Quantification was performed using a gallic acid standard curve; the results are expressed as gallic acid equivalents.

[0217] The total protein content was measured by spectrophotometry at 550 nm after colorimetric reaction of Biuret combined with Folin-Ciocalteu reagent. Quantification was performed using a standard BSA (Bovine Serum Albumin) curve.

[0218] The detailed composition of phenolic compounds was determined by liquid chromatography coupled with a UV detector fixed at 254 nm. The samples were separated on an UPTISPHERE CS EVOLUTION C18-AQ column using an Agilent 1200 HPLC system (Agilent Technologies). The mobile phases consisted of a 0.1% formic acid solution and methanol.

[0219] The molecular weight distribution profile of the proteins was determined by size exclusion chromatography coupled with a UV detector fixed at 254 nm. The samples were separated on a YMC Pack Diol 60 column (DL06503-2543WT). by an Agient 1200 HPLC system (Agilent Technologies). The mobile phase consisted of an aqueous solution of 0.05% NaN3, 0.2 M NaCl, 0.1 M phosphate buffer at pH 7.

[0220] The characterization and quantification of organic acids was performed by high-performance liquid chromatography coupled with mass spectrometry (Acquity Qda, Waters) equipped with a negative-mode electrospray ion source. Samples were separated on an EC 150 / 4.6 Nucleoshell RP 18plus-5pm column (Macherey Nagel: 763236.46) using an Agilent 1260 HPLC system (Agilent Technologies). The flow rate was 0.3 mL / min. The mobile phases consisted of a 0.01% formic acid solution and acetonitrile.

[0221] Results

[0222] The protein profile of the extracts is illustrated in Figure [Fig. 1]. The molecular weights of the conventional extract range from 7200 to 170 g / mol. In comparison, the autofermentation process of Example 1 broadens the molecular weight distribution of the extract proteins, with masses ranging from 15800 to 170 g / mol. Overall quantification analyses and comparisons of protein profiles thus highlight the increase in the amount of protein during the fermentation process compared to a conventional extract, particularly for masses above 1000 g / mol.

[0223] The extract from example 2 (auto-fermentation coupled with extraction) increases the complexity of the extract with the appearance of proteins with a molecular weight of around 19,000 g / mol and an increase in the signal for small molecules with a molecular weight of around 1000 g / mol.

[0224] The analysis highlights a decrease in the quantity of phenolic compounds during the self-fermentation process as illustrated in Tables [Table 1] and [Table 2].

[0225] The extract in Example 2 (autofermentation coupled with extraction) contains more phenolic compounds than a conventional extract (Example 5). Figure [Fig. 2] shows targeted consumption of complex phenolic compounds (flavonoids or glycosylated flavonoids) and the appearance of simple phenolic compounds such as phenolic acids during fermentation. Figure [Fig. 2] indicates a 54% increase in the presence of phenolic acids and an 87% decrease in the presence of flavonoids and glycosylated flavonoids in the autofermented jasmine extract compared to a conventional extract.

[0226] The self-fermented extract followed by an extraction (according to example 2) shows a 41% increase in phenolic acids than a self-fermented extract and 78% less flavonoids and glycosylated flavonoids than a self-fermented extract.

[0227] Analysis of the organic acids contained in the extracts, as illustrated in Figure [Fig. 3], shows a 42% increase in succinic acid and a 44% increase in the concentration of shikimic acid, which are specific compounds resulting from fermentation by microorganisms in self-fermented and self-fermented extracts followed by extraction versus a conventional extract.

[0228] [Table 1]: Analysis of compounds in jasmine flower extracts Conventional Extract Autofermented Extract Autofermented Extract + Extraction Phenolic Compounds (mg / kg) 783 630 1301 Protein (g / kg) 4.7 3.7 6.6 Shikimibic Acid (mg / kg) 135.7 243.2 241.4 Succinic Acid (mg / kg) 23.4 40.2 50.7

[0229] [Table 2]: Analysis of phenolic compounds from jasmine flower extracts (area under the curve) Conventional extract Autofermented extract Autofermented extract + Extraction Phenolic acids (mAU) 2304 5078 8749 Flavonoids and glycolicious flavonoids (mAU) 2606 329 71

[0230] Example 7: Analysis of violet flower extracts from examples 3 and 4 versus a conventional extract (example 5)

[0231] The analyses were carried out on the extracts of examples 3 and 4 before dilution and on a conventional extract of example 5.

[0232] Methodology

[0233] The total sugar content of the extract was determined by spectrophotometric assay. At 490 nm, colored complexes formed by sugars dissolved in sulfuric acid were reacted with phenol. The sugar content was determined using a standard glucose curve.

[0234] The detailed identification and quantification of the sugars present was carried out by high-performance liquid chromatography coupled to an ACQUITY Qda (WATERS) mass spectrometer equipped with an electrospray probe in negative mode. The samples were separated on a Luna Omega SUGAR column. 100A (Phenomenex: H21-180323) was analyzed using an Agient 1260 HPLC system (Agilent Technologies). The flow rate was 0.8 ml / min. The mobile phases consisted of a 20 mmol aqueous solution of ammonium acetate and 95 / 5 (ACN / H2O) 20 mmol of ammonium acetate.

[0235] The characterization and quantification of organic acids was carried out according to the method described in Example 6.

[0236] Results

[0237] The analyses highlight a consumption of 67% of the total sugars (illustrated [Fig.4]) and in particular the consumption of all the sugars involved in the fermentation processes (fructose and glucose) in the extracts of the invention (examples 3 and 4) compared to a conventional extract (example 5).

[0238] Analysis of the organic acids contained in the different extracts (illustrated [Fig. 5]) shows a 97% increase in the concentration of lactic and succinic acids and a 99% increase in propionic acid in the self-fermented violet extract compared to a conventional extract. The self-fermented extract followed by an extraction contains 11% more succinic acid and 15% more propionic acid than a self-fermented extract.

[0239] [Table 3]: Analysis of sugars and organic acids in extracts of violet flowers Conventional extract Autofermented extract Autofermented extract + Extract ion Total sugars (g / kg) 3.1 1.0 2.5 Fructose (mg / kg) 132.0 <LD <LD Glucose (mg / Kg) 243,0 <LD <LQ Acide lactique (mg / Kg) 19,2 552,7 544,8 Acide succinique ( mg / Kg) 6,3 235,1 263,6 Acide propionique (mg / Kg) <LQ 241,5 286,4

[0240] Example 8: Demonstration of the role of phytobiota in the self-fermentation of jasmine flowers (Jasminum grandiflorum)

[0241] Fresh jasmine flowers were sterilized by autoclaving for 10 minutes at 121°C to kill microorganisms. This plant material, devoid of phytobiota, was then used in the process described in Example 1 to constitute a control extract.

[0242] Physico-chemical analysis shows that the levels of phenolic compounds and proteins decrease sharply during the auto-fermentation process, whereas they are stable in the control condition.

[0243] These results demonstrate that the microorganisms of the phytobiota are directly responsible for the metabolic consumption of phenolic compounds and proteins during the self-fermentation process described in this application and in particular in Example 1.

[0244] [Table 4]: Comparison of the consumption of phenolic compounds and proteins in self-fermentation or control conditions (phytobiota killed) Extract of self-fermented jasmine flowers Extract of sterilized jasmine flowers Start of fermentation End of fermentation Start of fermentation End of fermentation Phenolic compounds (mg / kg) 912 486 1046 980 Protein (g / kg) 4.1 2.9 5 5.1

[0245] Example 9: In vitro evaluation of the hyaluronidase inhibitory capacity of fresh jasmine flower extracts obtained according to examples 1 and 5

[0246] Hyaluronidase catalyzes the degradation of hyaluronic acid (a glycosaminoglycan highly present in the dermis of the skin with hydrating and anti-aging properties) into mono or disaccharides as well as into smaller fragments of hyaluronic acid.

[0247] Hyaluronic acid has the ability to cause turbidity in the presence of an acidic albumin solution, which can be measured by spectrophotometer at 600 nm.

[0248] Protocol: The enzyme is incubated with the extracts obtained according to examples 1 and 5. The chosen concentration (vol / vol dilution) is prepared at a temperature of 37°C. The enzyme substrate, hyaluronic acid, is added to the mixture. The mixture is slowly homogenized and then incubated for exactly 45 minutes at 37°C. The hyaluronic acid remaining in the mixture is then contacted with an acidic albumin solution for 10 minutes before the transmittance is measured at 600 nM using a spectrophotometer.

[0249] The extract of self-fermented jasmine flowers prepared according to Example 1 exhibits a significantly stronger enzyme inhibitory power than the prepared extract conventionally according to example 5. The results are presented in [Table 5].

[0250] [Table 5]: Comparison of the percentages of hyaluronidase inhibition obtained in vitro between conventional and self-fermented jasmine extracts Conventional extract Self-fermented extract Extract concentration 0.5% 1% 2% 0.5% 1% 2% Hyaluronidase inhibition percentage 4 1 15 57.4 86.6 100

[0251] Example 10: In vitro evaluation of the tyrosinase inhibitory power of fresh violet flower extracts obtained according to examples 3 and 5

[0252] Tyrosinase is an enzyme involved in the synthesis of melanin produced by melanocytes in the skin. Its inhibition leads to a decrease in melanin production in the skin.

[0253] Protocol: The enzyme is incubated with the extracts obtained according to Examples 3 and 5 at a chosen concentration (vol / vol dilution) for 25 minutes at room temperature in a neutral pH buffer. Just before measuring fluorescence by spectrophotometry at 490 nm, the enzyme substrate, 2.5 mM L-tyrosine, is added to the mixture. The kinetics of the fluorescence emitted by the undegraded substrate are monitored for 45 minutes at 490 nm.

[0254] The self-fermented violet flower extract prepared according to Example 3 exhibits a significantly stronger inhibitory effect on the enzyme than the conventionally prepared extract according to Example 5, which shows no inhibitory effect on tyrosinase in vitro. Results are presented in [Table 6].

[0255] [Table 6]: Comparison of the percentages of tyrosinase inhibition obtained in vitro between conventional and self-fermented violet extracts Conventional extract Self-fermented extract 0.5% 1% 2% 0.5% 1% 2% Tyrosinase inhibition percentage 0 0 0 24.8 56.9 92.7

[0256] Example 11: Formula for a rich cream

[0257] [Tables?] Ingrédients (Nom de marque) INCI % w / w Phase A Eau purifiée Aqua Qsp 100 100 Optiphen™ Plus préservât ive Phenoxyethanol (and) Caprylyl Glycol (and) Sorbic Acid 1,50 Phase B Stabileze™ QM polymer PVM / MA Decadiene Crosspolymer 0,15 Phase C ProLipid™ 141 lamellargel Glyceryl Stéarate (and) Behenyl Alcohol (a nd) Palmitic Acid (and) Stearic Acid (and) Lecithin (and) Lauryl Alcohol (and) Myris tyl Alcohol (and) Cetyl Alcohol 5,00 Ceraphyl™ 494 ester Isocetyl Stéarate 4,00 Ceraphyl™ SLK ester Isodecyl Neopentanoate 4,00 DC580 Wax Stearoxytrimethylsilane (and) Stearyl Alco hol 2,00 Emulsynt™ GDL ester Glyceryl Dilaurate 3,00 Phase D Gransil DM-5 Dimethicone (and) Polysilicone-11 3,00 Phase E Hydroxyde de sodium Sodium Hydroxide 0,04 Eau purifiée Aqua 0,50 Phase F PF Bois Précieux Parfum / Fragrance 0,30 Unipure* Red LC 381 ADT-C CI 77491 (Iron oxides) (and) Isopropyl Tit anium Triisostearate (and) Bis-Hydroxyet hoxypropyl Dimethicone (and) PEG-2-Soy amine (and) Isophorone Diisocyanate 0,03 Phase G Extract according to example 2 propanediol (and) Jasminum Grandiflorum Flower Extract 2.00, Ronaflair Balance Gold CI 77891 (Titanium Dioxide) (and) Mica (and) Tin Oxide 0.30 Covabead Velvet 10 Polymethyl Methacrylate 1.00 Ronaflair Balance Red CI 77891 (Titanium Dioxide) (and) Mica (and) Tin Oxide 1.20 Phase H Aqua Purified Water 15.00 Natrosol™ Plus 330 CS HM HEC Cetyl Hydroxyethylcellulose 0.50

[0258] Preparation process

[0259] 1. Homogenize phase A in the main container and begin heating at 75-80°C;

[0260] 2. At 30°C, sprinkle into Phase B and homogenize while heating;

[0261] 3. In a separate beaker, prepare phase C, heat to 75-80°C until homogeneity;

[0262] 4. At 75°C, add phase C to the main container and homogenize for 10 minutes ;

[0263] 5. Allow the temperature to cool and add phase D at 65°C. Mix well to mix for 10 minutes;

[0264] 6. Premix phase E before adding it to the main container;

[0265] 7. Add phase E at 60°C. Mix well to homogenize for 10 minutes ;

[0266] 8. At 35°C, premix phase F before adding it and mix well;

[0267] 9. Premix phase G before adding it to the main container;

[0268] 10. Add phase G at 35°C. Mix well to homogenize;

[0269] 11. In a separate beaker, prepare phase H: sprinkle Natrosol™ into the water room temperature and homogenize while heating to 60°C;

[0270] 12. Add phase H at 30°C. Mix well to homogenize;

[0271] 13. Stop at 25°C.

[0272] The composition is thus presented in the form of a pink buttercream, with a pH between 4.90 and 5.40 and a viscosity (D0) of 160000 - 210000 cps (Brookfield RVT / Spindle D / 5 RPM / 1 minute / 25°C).

[0273] Example 12: Evaluation of the jasmine extract obtained according to example 2 on the expression of the piezolytic receptor in reconstructed epidermis

[0274] The proteins piezo2 and piezo2 have been identified as ion channels mediating mechanosensory transduction in mammalian cells (Coste, Bertrand et al. “Piezol and Piezo2 are essential components of distinct mechanically activated cation channels.” Science (New York, NY) vol. 330, 6000 (2010): 55-60). In the skin, keratinocytes participate in mediating tactile sensation by detecting and encoding this information for sensory neurons. Piezol is the main mechanotransducer of keratinocytes (Holt, Jesse R et al. Spatiotemporal dynamics of PIEZO1 localization Controls keratinocyte migration during wound healing. eLife vol. 10 e65415. 27 Sep. 2021).

[0275] Principle: The jasmine extract obtained according to Example 2 was evaluated for its ability to modulate the expression of the piezolytic mechanoreceptor in human skin ex vivo.

[0276] Protocol: Piezol receptor expression is assessed by indirect immunofluorescence on sections of reconstructed epidermis, pre-treated by topical application of the jasmine extract from Example 2 diluted to 2% (v / v) for 48 hours (once daily). Control reconstructed epidermis incubated in parallel under the same conditions receives placebo (Phosphate Buffer Saline, PBS). At the end of incubation, the reconstructed epidermis is fixed and paraffin-embedded for histological sectioning. Piezol receptor detection is performed by incubation with a primary anti-piezol antibody (Proteintech). After 1.5 hours of incubation followed by rinsing, the sections are incubated with a secondary anti-rabbit antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope).The expression of the piezolytic receptor is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0277] Results: As illustrated by Figure [Fig.6], when biopsies were treated with 2% jasmine extract, piezolytic receptor expression increased by 39%.

[0278] Conclusion: Jasmine extract showed a positive effect on piezolytic receptor expression.

[0279] Example 13: Evaluation of the jasmine extract obtained according to Example 2 on the expression of the oxytocin receptor (OXTR) on human skin biopsies

[0280] Principle: The aim of this experiment is to demonstrate an effect of jasmine extract on the synthesis of the oxytocin receptor in cultured human skin biopsies.

[0281] Protocol: Oxytocin receptor (OXTR) expression is assessed by indirect immunofluorescence on skin biopsies pre-treated by topical application of the jasmine extract from Example 2 diluted to 2% (v / v) for 48 hours (twice daily). Control biopsies incubated in parallel under the same conditions receive placebo (Phosphate Buffer Saline, PBS). At the end of incubation, the biopsies are fixed and paraffin-embedded for the performance of Histological sections. OXTR receptor detection is performed by incubation with anti-OXTR antibody (Proteintech). After 1.5 hours of incubation followed by rinsing, the sections are incubated with a secondary anti-rabbit antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). OXTR receptor expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0282] Results: As illustrated by Figure [Fig.7], when biopsies were treated with 2% jasmine extract, OXTR receptor expression increased by 63%.

[0283] Conclusion: Jasmine extract showed a positive effect on the expression of the olfactory receptor to oxytocin OXTR.

[0284] Example 14: Evaluation of the jasmine extract obtained according to Example 2 on the expression of oxytocin receptor (OXTR) messenger RNA in cultured keratinocytes

[0285] Oxytocin is a natural peptide that controls a wide range of specific actions in its target tissues, from cell growth and differentiation to reproduction and social behavior (Carter, C. Sue et al. “Is Oxytocin?” Nature's Medicine Pharmacological Reviews vol. 72, 4 (2020): 829–861). A large body of scientific literature supports the beneficial role of oxytocin in skin physiology and aging (Hayre, Nicole. “Oxytocin Levels Inversely Correlate with Skin Age Score and Solar Damage.” Journal of Drugs in Dermatology: JDD vol. 19, 12 (2020): 1146–1148). Other research also highlights the importance of the oxytocin receptor in preventing skin aging (Cho, SY et al. “Oxytocin alleviates cellular senescence through oxytocin receptor-mediated extracellular signal-regulated kinase / Nrf2 signalling.” The British journal of dermatology vol. 181,6 (2019): 1216-1225).

[0286] Principle: The aim of this experiment is to demonstrate an effect of jasmine extract on the synthesis of messenger RNA of the oxytocin receptor in human skin keratinocytes.

[0287] Protocol: The expression level of OXTR oxytocin receptor messenger RNA was assessed by qPCR (quantitative polymerase chain reaction) on cultured skin keratinocytes, pre-treated with jasmine extract obtained according to Example 2 for 48 hours at 1% (v / v dilution) in the culture medium (once daily). At the end of the culture period, the cells were lysed, the total RNA was extracted, and then converted to complementary DNA (cDNA) by reverse transcription. Quantification of OXTR receptor cDNA was performed by qPCR using a TaqMan probe for detection. The method of Quantification of delta-delta Ct allowed comparison of the difference in expression (ACt) between the gene of interest (OXTR) and the reference gene (GADPH).

[0288] Results: As illustrated by Figure [Fig.8], when cultured keratinocytes were treated with the jasmine extract obtained in Example 2 at 1%, the expression of OXTR receptor messenger RNAs was increased by 26%.

[0289] Conclusion: Jasmine extract showed a positive effect on the expression of OXTR receptor messenger RNAs.

[0290] Example 15: Evaluation of the jasmine extract obtained according to Example 2 on the expression of viperin in cultured keratinocytes

[0291] Viperin is one of the innate immune defense factors synthesized by keratinocytes (Garcia, Magali et al. “Innate Immune Response of Primary Human Keratinocytes to West Nile Virus Infection and Its Modulation by Mosquito Saliva.” Frontiers in cellular and infection microbiology vol. 8 387. 2 Nov. 2018). Viperin has been shown to catalyze the conversion of cytidine triphosphate (CTP) to 3'-deoxy-3',4'-didehydro-CTP (ddhCTP), a previously unknown ribonucleotide analog. The incorporation of ddhCTP causes the premature termination of RNA synthesis in some viruses (Gizzi, Anthony S et al. “A naturally occurring antiviral ribonucleotide encoded by the human genome.” Nature vol. 558,7711 (2018): 610-614.).

[0292] Principle: The aim of this experiment is to demonstrate an effect of jasmine extract on the level of viperin expression in cultured keratinocytes.

[0293] Protocol: Viperin expression was evaluated by indirect immunofluorescence on keratinocytes. Cultured keratinocytes were treated with jasmine extract obtained according to Example 2 at 1% (vol / vol dilution) for 48 hours (once daily). At the end of the culture period, the cells were fixed. Viperin detection was performed by incubation with an anti-viperin antibody (Sigma). After 1.5 hours of incubation followed by rinsing, the cells were incubated with a secondary anti-mouse antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The cells were then examined by epifluorescence microscopy (Zeiss Axiovert 200M microscope). Viperin expression was then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0294] Results: As illustrated by Figure [Fig. 11], the jasmine extract obtained according to Example 2 showed a positive effect of +40% on viperine expression in cultured keratinocytes.

[0295] Conclusion: Jasmine extract showed a positive effect on viperine expression.

[0296] Example 16: Evaluation of the jasmine extract obtained according to example 2 on the level of E-cadherin expression, on human skin biopsies pretreated with a blocker of piezolytic activity (Dookul)

[0297] Principle: The aim of this experiment is to demonstrate an effect of jasmine extract on the level of expression of E-cadherin, a molecule involved in tissue tone, in biopsies of cultured human skin.

[0298] Protocol: E-cadherin expression is assessed by indirect immunofluorescence on skin biopsies pretreated with a piezolytic blocker (Dookul), then treated by topical application of jasmine extract obtained according to Example 2 for 48 hours (twice daily), or with a piezolytic activator (Jedil) for 48 hours (once daily). Control biopsies incubated in parallel under the same conditions receive placebo (Phosphate Buffer Saline, PBS). At the end of incubation, the biopsies are fixed and paraffin-embedded for histological sectioning. E-cadherin detection is performed by incubation with anti-E-cadherin antibodies (Abcam). After an hour and a half of incubation, followed by rinsing, the sections are incubated in the presence of the anti-rabbit secondary antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen).The sections are then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). OXTR receptor expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0299] Results: As illustrated in Figure 10, when biopsies were pretreated with a piezolytic activity blocker (Dookul), a 44% decrease in E-cadherin expression was observed. When biopsies were treated with the piezolytic activator (Jedil), E-cadherin expression increased by 23%. When biopsies were treated with jasmine extract, E-cadherin expression increased by 95%.

[0300] Conclusion: The jasmine extract obtained according to Example 2 showed a positive effect on E-cadherin expression, mimicking the effect of the piezolytic receptor activator (Jedil).

[0301] Example 17: Evaluation of the jasmine extract obtained according to Example 2 on the level of expression of messenger RNA of the enzyme 11p-HSD1, in cultured keratinocytes

[0302] Principle: The aim of this experiment is to demonstrate an effect of jasmine extract on the level of expression of the messenger RNA of the lip-HSDl enzyme, also known as cortisone reductase, in cultured human skin keratinocytes.

[0303] Protocol: The expression level of messenger RNA of the enzyme 11[3-HSD1] is evaluated by qPCR (quantitative polymerase chain reaction) on Cultured human skin keratinocytes were pre-treated with jasmine extract obtained according to Example 2 for 48 hours at 1% in the culture medium (once daily). At the end of the culture period, the cells were lysed, and the total RNA was extracted and then converted to complementary DNA (cDNA) by reverse transcription. Quantification of the cDNA of the 11[3-HSD1] enzyme was performed by quantitative polymerase chain reaction using a TaqMan probe for detection. The delta-ΔCt quantification method allowed for comparison of the difference in expression (ACt) between the gene of interest (11(3-HSD1)) and the reference gene (GADPH).

[0304] Results: As illustrated by Figure [Fig. 11], when cultured keratinocytes were treated with the jasmine extract obtained according to Example 2 at 1%, the level of expression of the enzyme 11[3-HSD1] mRNA was decreased by -24%.

[0305] Conclusion: Jasmine extract showed an inhibitory effect on the expression level of the enzyme 11[3-HSD1.

Claims

Demands

1. A composition comprising an effective amount of fresh jasmine flower extract of the species Jasminum grandiflorum for its use in mitigating age-related decline in touch receptors (Piezol) and oxytocin receptors (OXTR), and improving in vivo well-being, wherein said extract is obtained by the following process: a) the plant material is mixed with water, b) the pH is adjusted, if necessary, to a value between 4 and 9, c) the mixture is kept under gentle stirring for a time between 15 and 30 hours, at a temperature between 30 and 40°C, at a pH between 4 and 7, to allow the auto-fermentation process, in a chamber allowing gas exchange with the atmosphere, to the auto-fermented plant material obtained in step c) phytic acid is added at a concentration between 1 and 5 mM, at a pH between 10 and 11,The pH of the resulting mixture is adjusted to a value between 6 and 8. The residual plant matter is then separated, the resulting liquid fraction is purified by successive filtrations to clarify the extract, and the extract is diluted with a physiologically acceptable solvent to obtain a diluted plant extract comprising, by weight of the total weight of the extract, 2 to 40 g / kg of dry extract, 0.2 to 30 g / kg of sugars, 10 to 4000 mg / kg of organic acids, 10 to 4000 mg / kg of phenolic compounds, and 10 to 2000 mg / L of amino acids.

2. Composition according to the preceding claim, wherein the composition comprises 0.05 to 5% of fresh jasmine flower extract of the species Jasminum grandiflorum, preferably 0.1 to 2.5 of said extract by weight relative to the total weight of the composition, and a physiologically acceptable medium.

3. A cosmetic composition comprising an effective amount of fresh jasmine flower extract of the species Jasminum grandiflorum for its use in improving the skin's immune defenses, wherein said extract is obtained according to the process comprising the following steps: a) the plant material is brought into contact with water, b) the pH is adjusted, if necessary, to a value between 4 and 9, (c) The mixture is kept under gentle stirring for 15 to 30 hours at a temperature of 30 to 40°C and a pH of 4 to 7 to allow for auto-fermentation, in a chamber allowing gas exchange with the atmosphere. To the auto-fermented plant material obtained in step (c), phytic acid is added at a concentration of 1 to 5 mM and a pH of 10 to 11. The pH of the resulting mixture is adjusted to a value of 6 to 8. The residual plant material is then separated. The resulting liquid fraction is purified by successive filtrations to clarify the extract and diluted with a physiologically acceptable solvent to obtain a diluted plant extract comprising, by weight of the total weight of the extract, 2 to 40 g / kg of dry extract, 0.2 to 30 g / kg of sugars, and 10 to 4000 mg / kg of organic acids, 10 to 4000 mg / kg of phenolic compounds and 10 to 2000 mg / L of amino acids.

4. A cosmetic composition for use according to claim 3, wherein the composition comprises 0.05 to 5% of fresh jasmine flower extract of the species Jasminum grandiflorum, preferably 0.1 to 2.5 of said extract by weight relative to the total weight of the composition, and a physiologically acceptable medium.

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