FR3046539A1 - ACTIVE PRINCIPLE OF PICHIA HEEDII AND COSMETIC USE IN PARTICULAR TO FIGHT THE EFFECTS OF POLLUTION ON SKIN - Google Patents

Abstract

The object of the invention is the cosmetic use of Pichia Heedii. The invention is directed in particular to an active ingredient of Pichia Heedii, in particular an active ingredient comprising glucomannans with molar masses of between 360 and 4000 Da, compositions including it and its cosmetic compositions, in particular anti-pollution.

Description

ACTIVE PRINCIPLE OF PICHIA HEEDDI AND COSMETIC USE IN PARTICULAR TO FIGHT THE EFFECTS OF POLLUTION ON THE SKIN

The present invention relates to the use in cosmetics of Pichia Heedii or an active ingredient of Pichia Heedii for its anti-pollution effect. The invention also relates to the active ingredients of Pichia Heedii, to cosmetic compositions including Pichia Heedii or an active ingredient of Pichia Heedii, and to a cosmetic treatment method for combating the effects of pollution on the skin.

As the number of urban dwellers continues to grow, many are looking for solutions to protect themselves from the impact of pollution. Indeed, beyond deleterious effects on health in general, it is now established that it is also harmful to the skin. Pollution is associated with skin aging, the appearance of pigmentary disorders and the emergence of pathologies such as atopic dermatitis or allergies (Krutmann J., Liu W., Li L., Pan X., Crawford M. , Sore G., Seite S. Pollution and skin: From epidemiological and mechanistic clinical studies, Journalof Dermatoiogicai Science, 76, 163-168 (2014), Vierkterter A., Schikowski T., Ranft U., Sugiri D., Matsui M., Kramer U., Krutmann J. Airborne Particle Exposure and Extrinsic Skin Aging, Journal of Investigative Dermatology, 130, 2719-2726 (2010)). Although this problem is universal, people in the Asia-Pacific region are particularly vulnerable. Pollutants are organic and non-organic substances. These include ozone, particulate matter (PM), polycyclic aromatic hydrocarbons (PAH), heavy metals, and cigarette smoke. Of a lipophilic nature, certain pollutants are able to penetrate inside the cells where they will be located at the level of the mitochondria. Their mechanism of action on the skin is not yet clearly elucidated. However, several potential pathways are suspected: - Alteration of the cutaneous bacterial flora: exposure to high concentrations of ozone results in a 50% reduction of the resident flora suggesting that it exerts a bactericidal effect. In addition, studies have shown that bacteria on the surface of the skin can partially metabolize pollutants. This incomplete degradation gives rise to toxic metabolites for the skin: - The induction of the inflammatory cascade: various pollutants have been described as associated with the production of inflammatory mediators. Among these mediators, there may be mentioned interleukins 1β, 8 and 6, and GM-CSF ("granulocyte macrophage colony-stimulating factor" or granulocyte-macrophage colony stimulating factor). This uncontrolled production in turn causes an alteration of epidermal differentiation. - Activation of the Aryl-hydrocarbon Aryl-hydrocarbon Receptor (AhR) receptor: The Aryl-hydrocarbon receptor is a cytoplasmic transcription factor that is able to detect the presence of pollutants. This receptor regulates several physiological functions among which cell proliferation, inflammation and melanogenesis. - Disturbance of the mitokines pathway (prohibitines): This still unexplored axis of research in cosmetics is dedicated to the sentinel function provided by the mitochondria, which allows the skin to remain connected to its environment. The mitochondrion captures the danger signal associated with a pollution episode and then transmits it to its partner organists through protein mediators: the mitokines. By this mechanism of novel intracellular communication, mitokines, especially prohibitin, allow the tissue to trigger an optimal antistress response. The objective of the invention is to prevent and combat the deleterious effects of pollution on the skin, by acting on several of these pathways, in particular on the Aryl-hydrocarcone receptor and the mitokines pathway, which constitutes a unpublished axis of action still unexplored in cosmetics.

To meet this objective, the invention proposes to use a particular yeast Pichia Heedii or an active ingredient of this yeast.

Yeast Pichia Heedii belongs to the Saccharomycetaceae family. It was found in Carnegiea gigantea cactus branches and trunks of the Pachycereus genus, family Cactaceae, in the Sonoran Desert in the southwestern United States in 1978.

Yeast Pichia Heedii is identified in the collections as ATCC numbers 34940, MUCL31256, CBS 6934; CCRC 21370; IFO 10019; UCD 76-503A.

Colonies are white to creamy, butyrous. They are convex, shiny to semi-glossy, and usually have a textured or irregular surface. After 3 days of culture, the cells are ovoid of 1.8-3.5 μm on 3.0-7.0pm. Ascospores are formed after 2-3 weeks of growth in the malt-agar medium. Pichia Heedii lives in strict aerobics.

Surprisingly, Pichia Heedii presents an "absolute" action against the deleterious effects of pollution on the skin. The use of this yeast or of an active ingredient derived from this yeast on the skin advantageously makes it possible to regulate at least two anti-pollution routes: the inactivation of the Aryl-hydrocarbon receptor in the face of pollutants; the restoration of the synthesis of mitokines, especially prohibitin, and the restoration of communication between the different cellular elements. The invention thus aims to use Pichia Heedii or an active ingredient of Pichia Heedii as a cosmetic active ingredient intended to be applied to the skin topically in a cosmetic composition, in particular to combat the effects of pollution on the skin. Advantageously, this use makes it possible to improve the barrier function as well as the color and luster of the skin. The invention also relates to a cosmetic active ingredient characterized in that it is an active ingredient of Pichia Heedii, in particular an active ingredient comprising glucomannans of molar masses between 360 and 4000 Da.

Finally, the invention also relates to a cosmetic composition including Pichia Heedii or an active ingredient of Pichia Heedii and a cosmetic treatment method. Other features and advantages will emerge from the detailed description of the invention which follows, with reference to the appended figures in which: FIG. 1 schematizes the activation or the inactivation of the Aryl hydrocarbon receptor (AhR) by a ligand and nuclear translocation leading to the regulation of target genes, - Figure 2 schematizes the subcellular locations of prohibitines in the absence of stress or in the presence of moderate stress, - Figure 3 represents the HPLC chromatographic profile of the active principle of the example 1, - Figure 4 represents the effect of an active ingredient according to the invention on the thickness of reconstructed epidermis transfected with siRNA PHB (corresponding to Table 9),

DEFINITIONS

For the purposes of the invention, the term "cosmetic active agent" or "cosmetic active ingredient" means at least one molecule, preferentially a set of molecules having an effect on the cells of the skin.

By "active principle of Pichia Heedii" within the meaning of the invention is meant any molecule or set of molecules derived from the structure of the Pichio Heedii ferment. These may be native ferment molecules or ferment molecules obtained by any type of transformation of the native molecules of the ferment, for example by hydrolysis. Preferably, it is a hydrolyzate of Pichia Heedii. The term "active principle" within the meaning of the invention excludes molecules produced by fermentation of Pichia Heedii.

By "hydrolyzate" is meant any active ingredient derived from Pichia Heedii, obtained by a process comprising at least one enzymatic or chemical hydrolysis step of Pichia Heedii. By "effects of pollution on the skin" within the meaning of the invention means the deleterious effects of pollution on the skin from a cosmetic point of view, namely in particular dehydration of the skin, a desquamation disorder. , development of acne, dull and gray complexion, premature aging and skin irritation.

"Pollution" means "environmental pollution", that is to say, all primary pollutants, directly from stationary sources, such as industry, heating, and mobile sources such as cars and airplanes, and secondary pollutants such as Ozone obtained from the transformation of primary pollutants under the action of solar rays and heat.

DETAILED DESCRIPTION OF THE INVENTION The invention therefore aims at the use of Pichia Heedii or an active ingredient of Pichia Heedii as a cosmetic active ingredient intended to be applied to the skin by topical application within a cosmetic composition. that is to say the cosmetic use of Pichia Heedii on the skin.

In particular, the invention relates to the use of Pichio Heedii or an active ingredient of Pichia Heedii to prevent and / or fight against the effects of pollution on the skin. To fight against the deleterious effects of pollution on the skin, Pichio Heedii or an active ingredient of Pichia Heedii acts mainly on two routes: - the Aryl-hydrocarbon route, and - the mitokine route.

With regard to the Aryl-hydrocarbon pathway, the Aryl hydrocarbon receptor (AhR) is able to detect the presence of pollutants and to reflect their biochemical and toxic effects.

It is activated by toxic molecules or particles emanating from cigarette smoke, incomplete incineration of waste or road traffic such as polycyclic aromatic hydrocarbons (PAHs), dioxins and its derivatives. Once activated, AhR is translocated from the cytoplasm to the nucleus where it regulates the expression of target genes including CYP1A1 (or Aryl-hydrocarbon hydroxylase). It is the prolonged induction of the AhR - CYP1A1 pathway through daily exposure to environmental pollutants that results in the deleterious effects of pollution. These effects result from the continuous and inappropriate production of pro-inflammatory mediators including cytokines, proteases and ROS, factors well known to cause the alteration of the different cellular components: DNA damage, lipid peroxidation and protein carbonylation (Kitamura M. , Kasai A. Cigarette smoke as a trigger for the dioxin receptor-mediated signaling pathway, Cancer Letters, 252, 184 - 194 (2007)). The pattern of activation of the Aryl hydrocarbon receptor (AhR) by ligand and nuclear translocation leading to the regulation of target genes is shown in Figure 1. The use of Pichia Heedii or an active ingredient of Pichia Heedii on the skin according to the invention makes it possible to deactivate the Aryl-hydrocarbon receptor and consequently to limit and fight against the deleterious effects of the pollution which result from the activation of this receptor. As regards the mitokine pathway, the invention acts in particular on prohibitines. Prohibitines are a family of proteins consisting of two homologs: prohibitin (prohibitin 1 or PHB1) and prohibitone (prohibitin 2 or PHB2). Their destinies are closely linked: the absence of one leading to the degradation of the other. In humans, although detectable in the bloodstream, prohibitines are mainly localized inside the cells, particularly in the inner membrane of the mitochondria where they form a large molecular complex. They play a key role in the development of mitochondria and their sustainability within the cell. They maintain the structure of the mitochondria, regulate its function, protect freshly synthesized respiratory chain proteins from uncontrolled degradation, and maintain the organization of mitochondrial DNA (mtDNA).

In response to stress, prohibitins are mobilized to other organelles in the cell to modulate various aspects of cell physiology. They are thus found at the level of the nucleus or the plasma membrane. In the nucleus, by interacting with various transcription factors such as NRF2 or PGCla, they will activate the anti-oxidant response and mitochondrial biogenesis, respectively, to protect the cell and enhance mitochondrial activity. Localized at the level of the cell membrane, they will maintain the structure of the lipid double-leaflet. Moreover, by interacting with cellular signaling proteins (MAPK, Raf / ERK, PI3K / Akt, etc.), they regulate the biological pathways involved in metabolism, migration, adhesion, proliferation and cell differentiation ( Mishra S., Ande SR, Nyomba BLG The role of prohibitin in cell signaling FEBS Journal, 277, 3937 - 3946 (2010) Zhou TB, Qin YH Signaling Pathways of Prohibition and Its Role in Diseases Journal of Receptors and Signal Transduction , 33, 28-36 (2013), Rajalingam K., Rudel T. Ras-Raf Signaling Needs Prohibitin Cell Cycle, 4,1503-1505 (2005)).

Prohibitines thus play the role of messenger by transmitting to the various elements of the cell the information relating to a potential danger. Because of their mobility, they are able to modulate the activity of the various organelles and on a larger scale the cell activity so that it is better equipped to respond to stress. Figure 2 shows the subcellular locations of prohibitines in the absence of stress or in the presence of moderate stress.

Thanks to the intracellular communication that they initiate in response to stress, prohibitines actively participate in the mechanisms of adaptation to adverse conditions: their cytoprotective role is undeniable.

According to the present invention, exposure of normal human keratinocytes to a polluted environment (lowering of oxygen content, exposure to benzo [a] pyrene or "particle matter") results in a significant reduction of prohibitin-1 synthesis. The use of Pichia Heedii or an active ingredient of Pichia Heedii on the skin according to the invention makes it possible to increase or restore the rate of prohibitines, in particular of prohibitin-1, in the cells of the skin, and allows thus to recreate an environment favorable to the communication between the organelles: essential element to the initiation of the machinery to fight against the stress created on the cells of the skin by the pollution. The invention therefore makes it possible to limit and combat the deleterious effects of pollution which result from the reduction of the rate of mitokines, in particular of prohibitin in cutaneous cells.

Thus, by renormalizing the synthesis of prohibitin and inactivating the Aryl-hydrocarbon receptor, Pichia Heedii or an active ingredient of Pichia Heedii has an antipollution effect. By regulating these two routes, the invention makes it possible in particular to: improve the color and brightness of the skin (increase of the radiation of the skin, reduction of the state of fatigue of the eyes, reduction of the synthesis especially the melanin of skin cells), - to improve the barrier function of the skin and to preserve the fragile skin of the pollution, in particular by improving the epidermic differentiation and reducing the insensible loss in water of the skin.

Pichia Heedii or an active ingredient in Pichia Heedii acts as a shield against the deleterious effects of environmental pollution. The invention makes it possible to obtain a skin that is more resistant, purified and a complexion that is fresh and soothed.

Preferably, the invention relates to the use of an active ingredient of Pichia Heedii, in particular of an active ingredient as described below.

Indeed, according to a second aspect, the invention relates to a cosmetic active ingredient characterized in that it is an active ingredient of Pichia Heedii.

The cosmetic active ingredient according to the invention comprises carbohydrates. These carbohydrates preferably represent at least 30% by weight relative to the total weight of dry matter of the active ingredient.

The carbohydrates of the active ingredient of Pichia Heedii according to the invention preferably have an average molar mass of between 360 and 4000 Da, more precisely between 500 and 100 ° C., preferentially 550 Da.

In a particularly suitable manner, the active ingredient is an active ingredient comprising in particular glucomannans with molar masses of between 360 and 4000 Da. Glucomannans are particular sugars, consisting of glucose and mannose. These sugars, with this particular size (molar masses between 360 and 4000 Da), are not all naturally present in Pichia Heedii and their production may require hydrolysis of Pichia Heedii.

Preferably, glucomannans with molar masses of between 360 and 4000 Da represent more than 50% by weight of the total carbohydrates of the active ingredient, more preferably more than 60%.

The determination of the molar masses is preferably carried out by steric exclusion chromatography. This method of liquid chromatography, with detection of refractive index (IR), makes it possible to separate the macromolecules as a function of their hydrodynamic volume.

The average molar mass corresponds to the average of the molar masses weighted by the intensity of the signals obtained for each molar mass.

The active ingredient may contain other molecules, including proteins and ashes. According to a particularly suitable variant, the active ingredient according to the invention is a hydrolyzate of Pichia Heedii, preferably an enzymatic hydrolyzate.

According to a suitable embodiment, the active ingredient useful according to the invention is in liquid form and has at least one of the following characteristics, preferably all: a solids content of between 13 and 57 g / l, preferably between and 38 g / l, - a content of carbohydrates of between 6 and 27 g / l, preferably between 12 and 18 g / l, that is between 30% and 70% by weight of dry matter,

Carbohydrates consist of glucose and mannose in simple form and in the form of glucomannans,

These carbohydrates are at least 50% by weight in the form of glucomannans with molar masses of between 360 and 4000 Da, the presence of peptides of less than 15% by weight relative to the weight of dry matter, an ash content of between 10 and 35% by weight relative to the weight of dry matter.

The solids content can be measured by passing the oven at 105 ° C. of a sample until a constant weight is obtained.

The overall content of carbohydrates can be determined by the DUBOIS method (Dubois M. et al., Analytical Chemistry, 28, 3, 350-356, 1956). The characterization of the molar mass of the carbohydrates present in the active principle of the present invention is carried out by HPLC method. The operating conditions are preferably the following:

Column: Polymer Laboratories (Varian) PLaquagel-OH 60, aquagel-OH 40 and aquagel-OH 30 size 300 x 7.5 mm (in series) with pre-column of the same characteristics Solvent: 0.3 M NaNO 3 buffer and NaH 2 PO 4 -2H20 0.01 M, pH7 in isocratic mode Detector: RI (Refractive Index)

The molar masses of the carbohydrates are evaluated by comparing the retention times of the peaks detected in the samples of the active ingredient with the retention times of previously injected standards.

The determination of simple sugars is carried out by ionic liquid chromatography.

The content of peptide compounds is determined by the method of LOWRY (Lowry et al., Protein measurement with the reagent reagent, J. Biol Chem, 193, 265, 1951).

The raw ash content can be determined by weighing the residues from the incineration of the samples of the active ingredient at 550 ° C in an electric muffle furnace. Instead of a liquid form, the active ingredient may also be in the form of atomized powder, for example.

The active ingredient according to the invention can be obtained by any process suitable for the transformation of Pichia Heedii molecules. Preferably, it is obtained by a process comprising a hydrolysis step, in particular enzymatic hydrolysis. According to a particularly suitable embodiment, the active principle is obtained by carrying out the following steps: Solubilization of Pichia Heedii powder in water, preferably at a rate of 20 g / l, Chemical and / or enzymatic hydrolysis , in order to hydrolyze carbohydrates, - Enzymatic inactivation by thermal treatment, - Separation of soluble and insoluble phases to recover the soluble phase, - Purification to eliminate molecules having a very high molar mass, - Concentration,

The parameters of the different steps must be adjusted in order to obtain active ingredients having the characteristics of the invention, in particular the presence of glucomannans having a molecular weight between 360 and 4000 Da. Some steps can be added in order to obtain a particular active ingredient, such as a step of: Clarification aimed at removing the compounds made insoluble during the heat treatment, this clarification can, for example, be carried out by filtration on a screen, Filtration final and sterilizing filtration intended to eliminate microorganisms that may be present in the product.

These steps taken individually are customary in the field of active extractions from plants or yeasts and the skilled person is able to adjust the reaction parameters on the basis of his general knowledge.

The active ingredient according to the invention or Pichia Heedii is preferably used in a composition, this composition comprising a cosmetically acceptable medium. These are compositions in different galenic forms, suitable for topical administration to the skin.

These compositions may especially 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 optionally microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersions, aqueous gels or powders. They can be more or less fluid and have the appearance of a cream, a lotion, a milk, a serum, an ointment, a gel, a paste or a foam, or in solid form.

It may be compositions comprising at least 0.25% of Pichia Heedii or of an active ingredient of Pichia Heedii according to the present invention, preferably between 0.25 and 10%. These compositions comprise, in addition to the active agent, a physiologically acceptable and preferably cosmetically acceptable medium, that is to say which does not cause unacceptable sensations of discomfort for the user such as redness, tightness or tingling.

The compositions according to the invention may contain as adjuvant at least one compound chosen from: oils, which may be chosen in particular from silicone oils, linear or cyclic, volatile or non-volatile; waxes, such as ozokerite, polyethylene wax, beeswax or carnauba wax, silicone elastomers, surfactants, preferably emulsifiers, whether they are nonionic, anionic or cationic or amphoteric, co-surfactants, such as linear fatty alcohols, thickeners and / or gelling agents, humectants, such as polyols such as glycerine, organic filters, inorganic filters, dyes, preservatives, fillers, tensors, sequestering agents, perfumes, and their mixtures, without this list being limiting.

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).

Of course, those skilled in the art will take care to choose any additional compounds, active or non-active, and their amount, so that the advantageous properties of the mixture are not, or not substantially, impaired by the addition envisaged.

These compositions are especially intended to be used to combat the deleterious effects of pollution on the skin. The invention also specifically relates to a cosmetic skin care method for combating the deleterious effects of pollution on the skin, in particular for increasing the effectiveness of the barrier function of the skin and / or improving the color and the skin. radiance of polluted skin, which consists in applying at least once a day to the skin of the face a composition comprising at least 0.10% by weight of dry matter of Pichia Heedii or of a cosmetic active ingredient of Pichia Heedii.

In order to illustrate these cosmetic effects on the skin, the following examples with their test results are presented.

EXAMPLES

Example 1 Active Principle According to the Invention

An example of a process for obtaining an active ingredient according to the invention comprises the implementation of the following steps: solubilization of Pichia Heedii powder in water at a rate of 20 g / l; enzymatic hydrolysis, in order to to hydrolyze carbohydrates, - Enzymatic inactivation by heat treatment for 15 minutes, - Separation of soluble and insoluble phases by decantation and recovery of the soluble phase, - Purification to remove substances having a molar mass greater than 10 kDa, - Filtration sterilizing on 0.22 μm.

The active ingredient obtained is a clear pale yellow liquid having a solids content of 32 g / l, a pH of 5 and a carbohydrate content (Dubois method) of 15 g / l, ie 47% of the dry matter. It also contains proteins. 68% of the carbohydrates of the active principle are glucomannans with molar masses of between 360 and 4000 Da. The determination of the molar masses is carried out by HPLC with RI detection. The HPLC / RI chromatographic profile of the carbohydrate moiety of this active ingredient is shown in Figure 3.

The carbohydrates are composed of 77% glucose and 23% mannose.

Example 2 Active Principle According to the Invention

An example of a process for obtaining an active ingredient according to the invention comprises the implementation of the following steps: solubilization of Pichia Heedii powder in water at a rate of 20 g / l, chemical hydrolysis, in order to to hydrolyze the carbohydrates, - Separation of the soluble and insoluble phases and recovery of the soluble phase, - Purification to remove substances having a molar mass greater than 10 kDa, - Sterilizing filtration over 0.22 pm

The active ingredient obtained is a clear pale yellow liquid having a solids content of 31 g / l, a pH of 5 and a sugar content (Dubois method) of 17 g / l, ie 55% of the dry matter. 60% of the carbohydrates of the active ingredient are glucomannans with molar masses of between 360 and 4000 Da.

Example 3 Use of an Active Principle According to the Invention in a Cream The composition has the following formulation:

This emulsion is white, supple shiny with a slight odor.

It has easy application, fast penetration and a dry finish. EVALUATION OF COSMETIC EFFICIENCY ACCORDING TO THE INVENTION Tests in vitro o Effect of an active ingredient of Pichia Heedii on the anti-pollution pathways Study of the mitokines pathway - 2D model The objective of this study is to evaluate the capacity of an active ingredient of Pichia Heedii to restore the mitokine pathway in a 2D model subjected to conditions mimicking a polluted environment.

For this, the action of an active ingredient of Pichia Heedii (example 1) on the synthesis of prohibitin was studied on normal human keratinocytes exposed: - to the causes of pollution, by applying two components of atmospheric pollution: benzo [a] pyrene (BaP) and fine particles (PM 2.5); - the consequences of the pollution on the oxygen content of the air, miming a depletion with a treatment with cobalt chloride.

Prohibitin synthesis was measured by Western blotting.

The operating protocol is described following.

Normal human keratinocytes are seeded in a suitable medium. The pollution with BaP is carried out by replacing the culture medium with a solution of benzo [a] pyrene (BaP).

To mimic pollution with fine particles, the culture medium is removed and the cells are attacked either with a solution of PM 2.5 fine particles or with a solution of cobalt. After several days, the cells are rinsed and then treated with the active ingredient of Pichia Heedii (Example 1) at 0.25%, 0.50% or 1.00%.

Then, the cell extracts are recovered and then stored at -80 °. The Western blot electrophoresis is carried out on precast TGX gel (Biorad).

The proteins are then transferred to PVDF membrane (Biorad). Immunolabeling is performed with the primary antibodies (anti-prohibitin antibody and anti-actin antibody), the secondary antibody and the revelation system.

The bands are visualized, semi-quantified by densitometry and then analyzed using the software.

The results obtained are presented in Tables 1 to 3.

The ability to restore prohibitines is calculated as follows:

Ability to restore Ratio test chemical compound - control ratio with chemical compound prohibitines = - x 100

Untreated control ratio - control ratio with chemical compound

Causes of pollution: aggression with benzofajpyrene:

*: significant result according to Student's t test / unpolluted control (p <0.05) **: significant result according to Student's t test / polluted control (p <0.10) *: significant result according to the t test Student / polluted control (p <0.05)

Table 1

Causes of pollution: aggression with fine particles:

*: significant result according to the Student t test / unpolluted control (p <0.05) **: significant result according to the Student t test / polluted control (p <0.10) A: significant result according to the t test Student / polluted control (p <0.05)

Table 2

Consequences of pollution: Reduction of oxygen content - Aggression with cobalt:

*: significant result according to the Student t test / unpolluted control (p <0.05) *: significant results according to the Student t test / polluted control (p <0.05)

Table 3

In a polluted environment, keratinocytes lose their ability to synthesize prohibitines (-12% with benzo [a] pyrene pollution, -15% with PM pollution, -37% with cobalt aggression).

An active ingredient of Pichia Heedii according to the invention is capable of restoring the mitokine pathway and consecutively allows the maintenance of intracellular communication: this phenomenon is essential for the initiation of an appropriate antistress response.

Tested at 1% on human keratinocytes subjected to a polluted environment, the active ingredient according to the invention significantly restores the synthesis of prohibitins by: - 67% against a benzo [a] pyrene attack; - 101% in the face of aggression to fine particles; - 68% against lowering of the oxygen content (cobalt).

Study of the Mitokine Pathway - 3D Model The objective of this study is to evaluate the ability of an active ingredient of Pichia Heedii according to the invention to restore the mitokine pathway in a 3D model subjected to conditions mimicking an environment polluted.

For this, the action of an active ingredient according to the invention (Example 1) on the synthesis of prohibitin was studied in reconstructed epidermis exposed to the consequences of pollution on the oxygen content of the air, mimicking depletion with cobalt chloride treatment.

Prohibitin synthesis was measured by Western blotting.

The operating protocol is described following.

Human keratinocytes are cultured in a specific culture medium. At J-1, the cells are exposed to a cobalt solution. At 0, the human keratinocytes treated or not treated with cobalt are inoculated on inserts and then incubated at 37 ° C.

The culture medium is changed every 2 days in the presence of the active ingredient of Example 1 at 0.10% and 0.25% (V / V).

After several days, the epidermal active principles are recovered and stored at -80 ° C. Electrophoresis is carried out on precast TGX gel (Biorad).

The proteins are then transferred to PVDF membrane (Biorad). Immunolabeling is performed with the primary antibodies (anti-prohibitin antibody and anti-actin antibody), the secondary antibody and the revelation system.

The results are given in Table 4.

The ability to restore prohibitines is calculated as follows:

*: significant result according to the Student t test / unpolluted control (p <0.05): significant result according to Student's t test / polluted treated control (p <0.10): significant result according to Student's t test / polluted treated control (p <0.05)

Table 4

Subject to an environment mimicking the consequences of pollution on the oxygen content of the air, the reconstructed epidermis show a synthesis of prohibitines significantly reduced by 28%.

These results show that in the face of a pollution episode, the active ingredient of Pichia Heedii according to the invention ensures the maintenance of intracellular communication and the triggering of a suitable antistress response by restoring the mitokines pathway. Tested at 0.25% on this model, the active principle of Example 1 significantly restores the synthesis of prohibitines by 58%.

Study of the Aryl-hydrocarbon Pathway The objective of this study is to evaluate the ability of an active ingredient of Pichia Heedii according to the invention to limit the activation of AhR following the exposure of human keratinocytes to an pollutant: benzo [a] pyrene (BaP). The effect on AhR activation has been studied by Western blotting.

The operating protocol is described following. At 0, normal human keratinocytes are seeded in a suitable medium.

After several days, the cells are attacked with a solution of benzo [a] pyrene (BaP) in the presence or absence of the active ingredient of Example 1 at 0.25%, 0.50% or 1.00%.

Then the cell extracts are fractionated and stored at -80 ° C.

Electrophoresis is carried out on precast TGX gel (Biorad).

The proteins are then transferred to PVDF membrane (Biorad). Immunostaining is performed with primary antibodies (anti-AhR antibodies and anti-TATA binding protein antibodies), secondary antibodies and a revelation system.

The bands are visualized, semi-quantified by densitometry and then analyzed using the software.

The results are shown in Table 5.

Each ratio is calculated according to the following formula:

*: significant result according to the Student t test / unpolluted control (p <0.05): significant results according to the Student t test / polluted control (p <0.05)

Table 5

In a polluted environment, the nuclear translocation of the AhR is significantly increased (+ 100%). Faced with a pollution episode, an active ingredient of Pichia Heedii according to the invention significantly reduces the activation of AhR.

Tested at 1%, the active principle of Example 1 significantly reduces this translocation by 105%. o Effect of an active ingredient of Pichia Heedii on the consequences of pollution on the skin: Study of the barrier function

In order to fight against the aggressions of the environment, the skin has a first line of defense, the epidermal barrier. This scaffold consists of keratinocytes coupled to a skeleton of proteinaceous nature. The objective of this study is to evaluate the ability of an active ingredient of Pichia Heedii according to the invention to promote the construction of a stratified and functional epidermis from human keratinocytes exposed to a depletion of the oxygen content ( consequences of pollution mimicked by cobalt treatment).

This epidermal construction was evaluated using: - Hematoxylin Eosin staining (HE) to measure the thickness of reconstructed epidermis obtained and evaluate their quality. - Differentiation markers: cytokeratin 10, filaggrin and loricrine to validate the functionality of reconstructed epidermis.

The operating protocol is described following.

Human keratinocytes are cultured in a specific culture medium. At J-1, the cells are exposed to a cobalt solution. At 0, normal human keratinocytes exposed to a cobalt solution are inoculated on inserts and then incubated at 37 ° C.

The culture medium is changed every 2 days in the presence of the active ingredient of Example 1 at 0.10% and 0.25% (V / V).

After several days, the reconstructed epidermis are recovered, fixed, dehydrated and included in paraffin. Sections (4pm) are then made using a microtome.

The Hematoxylin Eosin (HE) staining is then carried out.

Immunohistological markings of cytokeratin 10, filaggrin and loricrin are then made using primary antibodies (anti-cytokeratin 10 antibody, anti-filaggrin, anti-loricrin antibody), secondary antibodies.

Visualization of staining and immunostaining is performed using an image analysis system. The thickness of the epidermis was measured on the histological sections after HE staining. The syntheses of cytokeratin 10, filaggrin and loricrin are proportional to the intensity of the fluorescence (green color) present on the reconstructed epidermis.

Quantitative image analysis was performed using Matlab® software.

The results on the thickness of the reconstructed epidermis are presented in Table 6.

*: significant result according to Student's t test / unpolluted control (p <0.05) A: significant results according to Student's t test / polluted control (p <0.05)

Table 6

Faced with an episode of pollution, epidermal construction is significantly reduced by 56%.

Tested at 0.25% the active ingredient according to the invention restores the epidermal construction and allows the maintenance of a reconstructed functional epidermis.

The results on the syntheses of the differentiation markers are presented in Table 7.

Table 7

In a depleted oxygen environment, the reconstructed epidermis exhibits significantly decreased cytokeratin 10, filaggrin and loricrin synthesis (-68%, -99%, -68%).

These results show that the use of an active ingredient of Pichia Heedii makes it possible to compensate for the alteration of epidermal differentiation induced by environmental pollution and thus makes it possible to obtain a functional barrier.

Tested at 0.25% on this model, the active ingredient of Example 1 significantly restores the synthesis of these 3 proteins by 45%, 52% and 69% respectively. Characterization of the mechanism of action The objective of this study is the characterization of the mechanism of action of the active principle according to the invention and the evaluation of the role of prohibitin.

In order to answer this question, we have measured whether, in the absence of prohibitin (siRNA technology), an active ingredient of Pichia Heedii according to the invention (example 1) is capable of restoring the barrier function. The expression of prohibitin-1 (PHB) can be blocked using siRNA (siRNA-PHB) technology in a reconstructed epidermis model.

The siRNA-PHB human keratinocytes are made by transfection with siRNA-PHB. The reconstructed epidermis, negative control, siRNA-PHB and siRNA-PHB treated with the active principle according to the invention are obtained by seeding the human keratinocytes respectively negative control or siRNA-PHB. The culture medium is changed every 2 days in the presence or absence of the active ingredient of Example 1 at 0.25% (V / V).

After several days, the reconstructed epidermis are recovered, fixed, dehydrated and included in paraffin. Sections (4pm) are then made using a microtome. Expression of the prohibitin mRNA is determined. The RNAs were reverse transcripts and the resulting complementary DNAs were analyzed by the quantitative PCR technique. The mRNAs of the HPRT protein, an internal reference reference, were also analyzed in parallel with the prohibitin mRNA.

Quantification of fluorescence incorporation is measured continuously using a thermocycler.

The visualization is performed on a microscope coupled to an image analysis system.

The epidermal construction was evaluated using Hematoxylin Eosin (HE) staining, which makes it possible to measure the thickness of the reconstructed epidermis obtained and to evaluate their quality.

The results obtained on the study of prohibitin mRNA expression are presented in the following table:

Table 8

The reconstructed epidermis model siRNA-PHB is lacking synthesis of prohibitin.

The results of the effect on the thickness of reconstructed epidermis transfected with siRNA PHB are shown in Table 9 and Figure 4.

Table 9 Inhibition of prohibitin in human keratinocytes before epidermal construction leads to a decrease in the thickness of the reconstructed epidermis. These results show that in the absence of prohibitin, an active ingredient of Pichia Heedii according to the invention does not make it possible to reconstruct the barrier function. The effect of the active ingredient according to the invention is therefore prohibitin-dependent.

Effect of the active ingredient on the consequences of pollution on the skin: study on melanogenesis

Pollution has deleterious effects on the skin. It causes the appearance of pigmentary disorders that result from the activation of the pollutant receptor, the Aryl-hydrocarbon receptor (AhR). Indeed, AhR is expressed by melanocytes. Once activated by pollutants, it is translocated to the nuclear level where it will activate the genes involved in melanogenesis. The objective of this study is to evaluate the ability of an active ingredient of Pichia Heedii according to the invention to inhibit melanogenesis. The study was carried out on culture of melanocytes B16F1 or human melanocytes by measuring the level of melanin synthesized.

The operating protocol is described following. At 0, the melanocytes are seeded in a suitable medium.

The cells are then treated with the active principle of Example 1 at 0.25%, 0.50% and 1.00%.

The cells are trypsinized and counted.

The pellet is lysed with Triton-containing sodium phosphate buffer, centrifuged and recovered for melanin level determination.

The synthesized melanin is quantified by spectrophotometric reading at 490 nm, using a standard range of synthetic melanin.

The results obtained are shown in Table 10.

Table 10

It is found that an active ingredient of Pichia Heedii according to the invention reduces melanogenesis and can thus help the management of the deleterious effects of pollution on the pigmentation of the skin.

Indeed, tested at 1% on B16F1 melanocytes and human melanocytes, the active ingredient of Example 1 significantly reduces the melanin level by 38% and 43%.

In Vivo Tests o Description of the selected panel and the studies carried out The effect of an active ingredient of Pichia Heedii according to the invention formulated at 3% was evaluated on a Caucasian panel and on an Asian panel in comparison with a placebo on the following aspects: - Study of the effect on the preservation of the skin barrier - Study of the effect on the radiance of the complexion - Study of the lightening effect - Subjective evaluation

The Caucasian panel is composed of healthy volunteers, female, aged between 40 and 65 years old and selected according to a criteria of poisoned skin (high oxidized lipids, smoker and having a dull complexion).

The Asian panel is composed of healthy volunteers, female, aged between 40 and 65 years and presenting a dull complexion.

Studies on the Asian panel were conducted by the Engineering and Skin Biology Laboratory in Chengdu, Sichuan Province, China. The set of measurements carried out is: - Study of the insensible loss in water:

Measurements were made using a Tewameter® TM 300 (Courage & Khazaka). This device is equipped with a probe that measures the water vapor gradient between the skin surface and the ambient air, which gives information on the quality of the barrier function of the stratum corneum.

The measurements were performed on symmetrical areas in the cheeks.

A decrease of the PIE is characteristic of an improvement of the barrier function of the skin. - Study of the recovery of the barrier function:

The skin barrier at the forearms has been artificially attacked by successive applications of adhesive. These were applied to the surface of the skin, using an applicator delivering a constant pressure, and removed. The aggression was standardized at 15 g / hr / m2 which corresponds to a PIE of xerotic-skinned individuals (Berry N et al., Int J CosmetSci, 21, 241-252, 1999).

Water Insensitive Loss (PIE) measurements were performed using Tewameter® TM 300 (Courage & Khazaka), before (basal PIE) and immediately after aggression, as well as 1 day, 3 days, 4 days, 7 days, 10 days and 14 days after aggression. - Study of the radiance of the complexion: The evaluation of the radiance of the complexion on the Caucasian panel was carried out blind by two experts previously trained to judge different parameters representative of the radiance of the complexion. The evaluation is done from scales of scores (from 1 to 10) and the following parameters were retained:

The radiance of the skin, characteristic of a radiant complexion. The higher the intensity of the light hooks on the prominent areas of the face, the brighter the skin is. • The light pink color, which allows to characterize a radiant complexion. The more the complexion is pink, the more it is perceived as fresh. • The olive color is representative of the good-looking effect; if the olive color decreases the good-looking effect is more important. • The state of fatigue of the eyes, which allows to appreciate the general fatigue of the skin. The evaluation of these different parameters is done on the areas (cheekbones, forehead, chin, eyes).

The clinical scoring of skin color on the Asian panel was performed by an expert under standardized conditions, particularly in terms of volunteer position and ambient lighting.

During this evaluation, the expert scores the skin color according to 6 components (brown, light brown, beige, yellow, olive and light pink) evaluated as a percentage of saturation.

In addition, the expert evaluates the transparency, radiance and brightness of the skin on a scale of 0 to 100.

The transparency parameter corresponds to the degree of transmission of light through the skin surface. The higher this parameter, the more transparent the skin is. The brightness parameter evaluates the skin's ability to reflect light, the higher it is, the more the skin returns light.

The brightness parameter makes it possible to approach the notion of radiance of a healthy, healthy skin. The higher this parameter, the brighter the skin. - Evaluation of the clarity of the skin

Measurements of skin color were performed on the cheeks using a Chromameter® CM2600d (Konica Minolta). It converts colors in the range of human perception into a numeric code consisting of three parameters: • L *: represents clarity (from dark to pale) • a *: represents the range from green to red • b *: represents the range from blue to yellow a * and b * are chrominance parameters and L *, a luminance parameter.

The parameter ITA (Individual Typological Angle) allows to evaluate the variation of the brilliant aspect of the skin. This parameter defines the degree of pigmentation of the skin by integrating the clarity (L *) and the melanization parameter (b *) according to the following formula: ITA ° = [Arc tan ((L * -50) / b *)] x 180 / π The higher the ITA parameter, the more the skin appears clear and luminous.

The parameters L * and ITA have been retained to study the effect of an active ingredient according to the invention on the lightening of the skin. Subjective evaluation The subjective evaluation by the Caucasian panel of the sensations observed after 28 days of twice-daily treatment with a composition containing the active ingredient according to the invention and the placebo was evaluated using self-evaluation questionnaires containing the closed questions:

The subjective evaluation by the Asian panel of the sensations felt after 28 days of twice-daily treatment with a composition containing the active ingredient according to the invention and the placebo was evaluated using self-evaluation questionnaires containing the so-called closed questions. :

The results of the closed questions were analyzed by combining the "somewhat agree" and "agree" responses. o Efficiency tests

Study of the effect of an active ingredient of Pichia Heedii to reinforce the cutaneous barrier. Measure of the PIE. The objective of this study was to evaluate, in vivo, the restructuring effect of an active ingredient according to the invention formulated at 3% in emulsion on the barrier function of the skin.

This study was conducted on 20 Caucasian volunteers, healthy, female, middle age 53 + 2 years, selected.

Measurements of the insensitive loss in water of the face were made before and after 14 and 28 days of twice-daily applications in the hemi-face of the placebo and of the active principle (example 3) according to the invention. The average of the results obtained is shown in Table 11.

Table 11

It is found that under the conditions of this study, from 14 days of twice-daily applications and in comparison with placebo, an active ingredient of Pichia Heedii according to the invention formulated at 3% in emulsion, significantly reduces the insensible loss of water in the skin. 8.4% (p = 0.0396).

This effect continues after 28 days of twice-daily application to achieve a significant decrease in water-insensitive loss of 11.2% (p = 0.0012) on artificially unimpaired skin.

This effect was observed in 80% of the volunteers.

An active ingredient of Pichia Heedii according to the invention thus improves the barrier function of the skin.

Effect of the active substance of Pichia Heedii on the capacity of recovery of the cutaneous barrier after artificial alteration (strippings). Measure of the PIE. The objective of the study was to evaluate, in vivo, the effect of the active ingredient of Pichia Heedii according to the invention formulated at 3% gel-emulsified on the ability to recover the barrier function of the skin after a single attack by stripping.

This study was conducted on 20 Caucasian volunteers, healthy, female, middle age 56 + 2 years. The effect of an active ingredient according to the invention on the skin barrier recovery capacity following a single attack by stripping was studied after a pretreatment of 28 days of twice-daily placebo applications and an active ingredient of Pichia Heedii (example 3), at the inner side of the forearms.

A summary of the results is presented in Table 12.

Table 12

Under the conditions of this study, after a pretreatment of 28 days in twice-daily applications and in comparison with placebo, an active ingredient of Pichia Heedii according to the invention formulated at 3% gel-emulsified allows the skin a recovery of the barrier function faster following a single attack by stripping.

Indeed, after a single aggression, the recovery of the barrier function is practically total with the invention (91%) in only 4 days (96h), unlike the placebo treated area which required 7 days before observing a total recovery. The recovery on the zone treated with the invention is significantly greater than that on the placebo zone at 24h and 72h (respectively p = 0.0584 and p = 0.0142).

An active ingredient of Pichia Heedii according to the invention formulated at 3% thus improves the recovery capacity of the barrier function of the skin.

Study of the effect of Pichia Heedii on the radiance of the complexion • Study on a Caucasian panel The objective of this study was to evaluate, in vivo, the effect of an active ingredient of Pichia Heedii according to the invention Formulated to 3% emulsion on the radiance of the complexion of the skin against placebo. This study was performed on 20 Caucasian volunteers, healthy, female, average age 53 + 2 years. The evaluation of the radiance of the facial complexion was performed blindly by two trained experts before and after 14 and 28 days of twice-daily applications of the placebo and the invention (Example 3).

A summary of the results is presented in Table 13.

Table 13

It is found that under the conditions of this study, from 14 days of twice-daily applications and in comparison with placebo, an active ingredient of Pichia Heedii formulated at 3% in emulsion: significantly improves the radiation of the skin by 7% (p = 0.0184) and the pink color, characteristic of a fresh complexion, of 14% (p = 0.0019). significantly decreases the olive color by 7% (p = 0.0430) and the eye fatigue state by 7% (p = 0.0378).

This effect continues after 28 days of applications, in fact an active ingredient of Pichia Heedii according to the invention: significantly increases the radiation of the skin by 12% (p = 0.0092, effect validated in 80% of the volunteers) as well as the pink color of 17% (p = 0.0002, validated effect in 80% of volunteers). - allows a significant decrease in the olive color of 15% (p = 0.0011) and the eye fatigue state of 14% (p = 0.0047). This effect was observed in respectively 65% and 80% of the volunteers. • Study on an Asian panel The objective of this study was to evaluate, in vivo, the effect of an active ingredient of Pichia Heedii formulated at 3% in emulsion on the radiance of the complexion of Asian subjects. 28 healthy female volunteers, mean age 50 + 5 years were included in this study. The evaluation of the radiance of the complexion of the face was performed blind, by a trained expert, before and after 14 and 28 days of twice-daily applications of the placebo and the invention (example 3) in half-face.

A summary of the results is presented in Table 14.

Table 14

Under the conditions of this study, after 28 days of twice-daily application and in comparison with placebo, we find that an active ingredient of Pichia Heedii formulated at 3% in emulsion: * Significantly decreases the colors of the skin of people of origin Asian clinically evaluated. Indeed, the invention decreases: - the brown color of 9% (p = 0.031), this effect was observed in 61% of volunteers; - the light brown color of 8% (p = 0.013), this effect was observed in 68% of volunteers; - the olive color of 7% (p = 0.064), this effect was observed in 61% of the volunteers; - the yellow color of 12% (p = 0.006), this effect was observed in 68% of volunteers. In addition, the invention significantly improves the light pink color by 24% (p <0.001). Finally, the invention improves the parameters relating to the reflection of light, by increasing: the brightness of the skin by 7% (p = 0.250); skin radiation of 6% (p = 0.264); - the transparency of the skin of 6% (p = 0.115).

Thus, an active ingredient of Pichia Heedii according to the invention improves the radiance of the complexion of people of Asian origin.

Effect of Pichia Heedii on the lightening of the skin of Asian subjects. Colorimetric study. The objective of this study was to evaluate, in vivo, the effect of the active ingredient according to the invention formulated at 3% in emulsion against placebo on the lightening of the skin of Asian subjects. 28 healthy, female volunteers, mean age 50 + 5 years, were included in this study. The measurements were performed blindly to the face using a Chromameter® before and after 14 and 28 days of twice-daily application of the placebo and the active ingredient (example 3) in half-face.

A summary of the results is presented in Table 15.

Table 15

Under the conditions of this study, after 28 days of twice-daily applications and in comparison with placebo, the active ingredient according to the invention formulated at 3% emulsion tends to lighten the skin of people of Asian origin.

Indeed, a significant increase in L * and ITA ° parameters, characteristic of lighter and brighter skin, is observed after 28 days of twice daily application of the active principle.

Subjective evaluation of the effect of the active principle according to the invention Pichia Heedii • Study on Caucasian panel The objective of this study was to quantify the sensations observed during the twice-daily use of the active ingredient according to the invention formulated at 3% in emulsion (example 3) and tested against placebo for 28 days as a twice-daily treatment in a half-face.

This study was performed on 20 Caucasian volunteers, healthy, female, average age 53 + 2 years, selected according to the criteria described.

The results of the closed questions are shown in Table 16.

Table 16

Under the conditions of this study, after 28 days of twice-daily applications and in comparison with placebo: • the active ingredient makes the skin more radiant for 94% of the volunteers, compared to 74% for the placebo (p = 0.1065). • 94% of them find that the active ingredient makes their complexion look cooler, compared to 74% for placebo (p = 0.0459). • For respectively 83% and 74% of volunteers, the active ingredient purifies the skin and provides a revitalizing effect, compared to 63% for placebo (respectively p = 0.0337 and p = 0.1003). • In addition, for 68% of volunteers, the active ingredient makes the skin more resistant, 61% make the same finding after 28 days of placebo applications (p = 0.0484). • Finally, all volunteers found that with the active ingredient, their complexion is sobered, against 84% for placebo (p = 0.0821).

After 28 days of twice-daily application, the active ingredient formulated at 3% emulsion is generally perceived as more effective than placebo.

In fact, most of the volunteers found their skin more resistant and purified, as well as their complexion, which was cooler and softer. • Asian panel study The aim of this study was to quantify the sensations observed during the twice-daily use of the active ingredient formulated at 3% emulsion (example 3) and tested against placebo for 28 days in twice-daily face. 28 Asian volunteers, healthy, female, mean age 50 ± 5 years, selected according to defined criteria. The results of the closed questions are shown in Table 17.

Table 17 • The active ingredient-treated side makes the skin brighter and refines the skin texture for 93% of the volunteers, compared to 79% and 89% respectively for the placebo-treated side (respectively p = 0.1335 and p = 0.6527). • 89% of them agree that with the active ingredient the complexion appears to be sobered and the skin revitalized, compared to 86% and 79% respectively for the side treated with the placebo (respectively p = 0.6993 and p. = 0.2851). • For 96% of volunteers, the active ingredient purifies the skin, and evenifies the complexion. They are 82% to make the same finding on the side treated with the placebo (p = 0.0896). • 96% find that the active ingredient reduces signs of fatigue compared to 86% for placebo (p = 0.1686). • In addition, all volunteers find that with the active ingredient their skin looks smoother. 89% report the same after 28 days of placebo (p = 0.0813). • Finally, for 97% of volunteers, the active ingredient reduces imperfections, compared with 82% for placebo (p = 0.2352).

After 28 days of twice-daily application, the active ingredient formulated at 3% emulsion is generally perceived as more effective than placebo.

Indeed, the volunteers of Asian origin found mostly their complexion more uniform, their skin more luminous and smooth.

Claims (22)

1. Cosmetic active ingredient characterized in that it is an active ingredient of Pichia Heedii. 2. Cosmetic active ingredient according to claim 1, characterized in that it comprises at least 30% of carbohydrates by weight relative to the total weight of dry matter of the active ingredient. 3. Cosmetic active ingredient according to claim 1 or 2, characterized in that it comprises glucomannans of molar masses between 360 and 4000 Da. 4. Cosmetic active ingredient according to the preceding claim, characterized in that the glucomannans of molar masses between 360 and 4000 Da represent at least 50% by weight of the carbohydrates of the active principle. 5. Active ingredient according to one of the preceding claims, characterized in that it has a solids content of between 13 and 57g / l. 6. Active ingredient according to one of the preceding claims, characterized in that it has a solids content of between 26 and 38g / l. 7. Active ingredient according to one of the preceding claims, characterized in that it is a hydrolyzate Pichia Heedii. 8. Active ingredient according to one of claims 1 to 7 for use in combating the deleterious effects of pollution on the skin by renormalizing the synthesis of prohibitin-1. 9. Active ingredient according to one of claims 1 to 7 for use in combating the deleterious effects of pollution on the skin by inactivating the Aryl-hydrocarbon receptor (AhR). 10. Use of Pichia Heedii or an active ingredient of Pichia Heedii as a cosmetic active ingredient for application to the skin for topical application in a cosmetic composition. 11. Use according to claim 10, for a cosmetic anti-pollution effect on the skin. 12. Use according to one of claims 10 or 11, to combat the deleterious effects of pollution on the skin. 13. Use according to one of claims 10 to 12, to improve the color and luster of the skin. 14. Use according to the preceding claim, for improving the color of the skin by reducing the melanin synthesis of the skin cells. 15. Use according to one of claims 10 to 14 for improving the barrier function of the skin. 16. Use according to the preceding claim, for improving the barrier function of the skin by improving epidermal differentiation. 17. Use according to the preceding claim, for improving the barrier function of the skin by reducing the insensible loss of water in the skin. 18. Use according to one of claims 10 to 17, characterized in that the active ingredient of Pichia Heedii is an active ingredient according to one of claims 2 to 7. 19. Cosmetic composition, characterized in that it comprises at least 0.25% by weight of dry matter of Pichia Heedii or an active ingredient of Pichia Heedii. 20. Cosmetic composition according to claim 19, characterized in that the active ingredient of Pichia Heedii is a cosmetic active ingredient according to one of claims 2 to 7. 21. A cosmetic skin care method for combating the deleterious effects of pollution on the skin, characterized in that it consists in applying at least once a day to the skin of the face a composition according to one of the claims. 19 to 20, 22. Cosmetic skin care method according to claim 21, for increasing the effectiveness of the barrier function of the skin and / or improving the color and luster of polluted skin.