FR2973699A1 - NOVEL ACTIVE INGREDIENT TO STIMULATE THE PRODUCTION OF ANTI-MICROBIAL PEPTIDES IN SCALP, USEFUL IN PREVENTING AND / OR TREATING SCALPLET FILM CONDITIONS - Google Patents

Abstract

The present invention relates to the cosmetic use of a lysate of bacteria (s) in a complete fermentation medium, said lysate in its complete fermentation medium being capable of inducing or stimulating the production of antimicrobial peptides in the scalp, as an active ingredient for preventing and / or treating dandruff conditions of the scalp, including dandruff scalp conditions associated with a prevalence of pathogenic microorganisms on the scalp and / or ecofloral imbalance of the scalp. .

Description

The invention relates to the cosmetic use of a bacterial lysate (s) in a complete fermentation medium, said lysate in its complete fermentation medium being capable of inducing or stimulating the production of at least one antimicrobial peptide at the level of the scalp, as an active ingredient to prevent and / or treat dandruff conditions of the scalp, including dandruff conditions associated with a proliferation of pathogenic microorganisms on the scalp and / or an imbalance of the ecoflora of the scalp. The skin is a tissue whose cells are contiguous and integral with each other. The cutaneous tissue forms an outer covering comprising sebaceous or sweat glands, and hair follicles. The skin, and especially the scalp, are epithelia with continual renewal. Renewal, or desquamation, is a coordinated and finely regulated process resulting in the elimination of surface cells, insensitively and nonvisibly. However, abnormal or irregular desquamation of the stratum corneum cells, for various reasons, can lead to the formation of large, thick, visible to the naked eye cells called "dander" or "dandruff" as part of the scalp, or in other situations, thinning of the stratum corneum. The desquamation disorders, resulting from abnormal or irregular desquamation, can lead to a fragility, even a defect of the barrier properties of the epidermis. As examples of factors promoting the development of dander or dandruff, mention may be made of stress, the winter period, an excess of sebum, a lack of hydration, or the colonization of the skin or hair follicles by the yeast Malassezia sp. These factors have, in particular, as a common feature of causing and / or promoting an inflammatory skin condition. Such inflammation enhances the appearance or even increases the presence of dander or dandruff. In particular, yeasts of the Malassezia sp. Type, constituting a part of the commensal flora normal to the surface of the scalp in subjects without film, see their proportion increase substantially in case of dandruff, or in case of associated seborrheic dermatitis. The imbalance of the ecoflora of the scalp is a factor favoring or even reinforcing the presence of dandruff. The presence of dander or dandruff can be chronic, frequent, recurrent, and socially disabling conditions because of their obvious unsightly nature. Moreover, the dandruff condition of the scalp or the abnormal desquamation of the skin can result in an alteration of the barrier function of the epidermis, or generate sensations of itching or pruritus, leading to scraping behaviors amplifying the phenomenon the appearance of scales or dandruff and, in turn, irritation of the scalp or skin. The dandruff conditions of the scalp may be of the fatty type or of the dry type. The dry dandruff conditions of the scalp are more frequently manifested, and are amplified, during disorders of the hydration of the skin, and in particular during severe dryness of the epidermis of the scalp. Also, since the scalp is rich in sebaceous glands, a dandruff condition can develop more easily in the presence of excessive sebum and be more easily pruriginous.Thus, excessive secretion of sebum, or hyperseborrhoea, promotes the appearance of a condition. Fat dandruff of the scalp, or fatty dandruff, generally associated with discomfort, sensations of discomfort, aesthetic disorders, even skin pathology.The dandruff states generally respond to different local or systemic treatments. effectiveness of these treatments is only suspensive and involves a rigorous follow-up on the part of the user (frequency of use and sufficient time of application), but a daily and long-term use of these treatments can cause a phenomenon addiction reducing their effectiveness, habituation usually being associated with a rebound phenomenon occurring on the This phenomenon is manifested by hyperseborrhoea, aggravating the dandruff condition and altering the barrier function of the scalp. Furthermore, the aggressiveness of certain antidandruff active against epidermal cells or ecoflora of the scalp can also affect the barrier functions of the latter and cause worsening of the dandruff condition. Finally, the effectiveness of anti-dandruff treatments is often slow to develop and requires a rigorous application in the long term. This latency often leads to a failure to monitor treatment. As a result, many failures occur in the implementation of these treatments. Many epidermal factors, whose expression, biological activity or maturation are altered, diminished or increased, are known to be involved, directly or indirectly, in the process of renewal, or desquamation of the scalp.

These factors can be used as biomarkers of the scalp, as screening targets, or even as cosmetic assets. However, there are still many unknowns as to the intimate mechanism and factors involved in desquamation of the skin, and in particular in the appearance of dandruff.

There is still a need for new assets or new treatments to prevent and / or treat a disorder of skin desquamation, and more particularly a dandruff condition of the scalp.

There is also a need for new cosmetic treatments to prevent, reduce and / or treat scalp dandruff conditions that are effective and have no side effects that may affect good compliance. There is still a need for a cosmetic treatment of dandruff conditions of the scalp does not affect the ecoflora of the scalp, or even strengthening the presence of a healthy ecoflora. There is a need to have a cosmetic treatment of the dandruff states able to maintain or even strengthen the barrier properties of the scalp. There is a need for cosmetic treatments of dandruff states which are free of the aforementioned side effects, and in particular which do not induce hyperseborrhea, seborrheic dermatitis or itchy conditions. There is also a need for a cosmetic treatment of the dandruff states which does not induce inflammatory conditions. The object of the present invention is to satisfy these needs. The present invention aims to meet these needs and aims in particular to provide a new asset meeting the above requirements and demonstrating in particular an efficiency with regard to disorders of the barrier function of the scalp facing a pathogenic microflora. As described in detail below, the new active ingredient of the invention is advantageously used in the implementation of cosmetic treatment processes of dandruff conditions of the scalp.

More specifically, the present invention relates to the cosmetic use of a bacterial lysate (s) in a complete fermentation medium, said lysate in its complete fermentation medium being capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp, as an active ingredient for preventing and / or treating dandruff conditions of the scalp, particularly associated with an imbalance of the ecoflora of the scalp. The prevention and / or treatment of dandruff states includes prevention and / or treatment of dandruff and / or prevention and / or treatment of scalp itching related to the presence of dandruff.

Collectively, the antimicrobial peptides consist of peptides, many of which are already known in the art, possessing directed activity against various microorganisms, including bacteria, fungi and protozoa. In general, the antimicrobial peptides have both a hydrophilic and a hydrophobic moiety, which peptides are therefore soluble in an aqueous environment and are simultaneously capable of penetrating through the lipid membranes. The ability of these peptides to penetrate through the lipid membranes gives them the ability to cross the microbial membrane walls (including bacterial, fungal and protozoan) to exert their microbicidal activity.

Antimicrobial peptides are produced in the scalp, both in the healthy scalp and in the scalp affected by a physiological disorder, said physiological disorder may be due to endogenous causes (eg imbalance or metabolic deficit of the cells scalp) or exogenous causes (mechanical injury (eg injury or surgical treatment), overexposure to UV radiation, burning, abrasion of the dermis by exposure to chemical compounds, trauma caused by regenerative treatment at the same time). using a laser, etc.). It is generally accepted that the presence of antimicrobial peptides in the cutaneous tissue, including the scalp, contributes to the barrier function of said cutaneous tissue, in addition to the mechanical contribution made by the striatum corneum.

In fact, numerous scientific reports have now established that the barrier function of cutaneous tissue, including scalp, is composite and also includes, in addition to the mechanical contribution of the striatum corneum and the chemical contribution of the antimicrobial peptides, also a contribution immune to both nonspecific (macrophages, polynuclear neutrophils) and specific (antibody production).

The normal presence of antimicrobial peptides in the scalp thus contributes to the maintenance of a healthy scalp, and in particular a scalp free of dandruff. As already indicated above, it has been shown according to the invention that the cosmetic use of an active ingredient derived from a bacterium, said active agent being capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp, can be used successfully for the prevention and / or treatment of dandruff conditions of the scalp.

According to the invention, an active agent capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp is capable of maintaining or restoring the production of said at least one antimicrobial peptide in the scalp. For the purposes of the present invention, the term "antimicrobial peptide" includes any peptide having antibacterial, antifungal or antiprotozoal activity produced in the scalp. The antimicrobial peptides include defensins, cathelicidines and dermicidine. Preferably, the antimicrobial peptides whose production induction or induction of production is targeted by the invention are defensins. Defensins consist of cationic peptides containing conserved cysteine-rich units. There are three defensin subfamilies, alpha defensins, beta defensins and theta circular defensins, respectively. Alpha and beta defensins are distinguished by the location of disulfide bridges. Even more particularly, the antimicrobial peptides whose production induction or induction of production is targeted by the invention are beta defensins. Human beta defensins 1 to 4 are expressed in keratinocytes. Among the antimicrobial peptides whose induction of production or the induction of production is sought by the invention, preference is given to defensin beta 2/4 and Rnase 7. Within the meaning of the invention, an active agent capable of in inducing or stimulating the production of at least one antimicrobial peptide, which is derived from a bacterium, includes (i) the active agents inducing a detectable increase in the expression of at least one gene encoding an antimicrobial peptide. Thus, the active agents capable of inducing or stimulating the production of at least one antimicrobial peptide include the active agents inducing a detectable increase in the expression of a gene selected from (i) the gene encoding the beta 2/4 defensin and ii) the gene encoding Rnase7. In order to determine whether a given active agent is capable of inducing or stimulating the expression of at least one antimicrobial peptide in the scalp, a person skilled in the art can test the ability of said active agent to increase the expression of at least one coding gene. an antimicrobial peptide, in particular the gene encoding beta4 defensin or the gene encoding Rnase7, after bringing said active agent into contact with human keratinocytes in culture, as illustrated in the examples, to determine the level of expression of a gene coding for an antimicrobial peptide, a quantitative PCR amplification technique (Q-PCR) is preferably used, as illustrated in Example 3. According to the invention, the induction of the expression of a gene coding for an antimicrobial peptide is detectable when the signal generated due to the presence of cDNA corresponding to the messenger RNA transcribed from said gene is greater than four times the background noise. According to the invention, the stimulation of the expression of a gene encoding an antimicrobial peptide is detectable when the signal generated due to the presence of cDNA corresponding to the messenger RNA transcribed from said gene, in an individual to whom administered an active ingredient according to the invention, is at least 30% greater than the corresponding signal generated in an individual who has not received said active agent according to the invention, or in the same individual before administration of said active agent. of the invention, said bacterial-derived active agents consist of a lysate in its complete fermentation medium, which is capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp. For the purposes of the present invention, the expression "lysate in a complete fermentation medium" means that the lysate is used and is present in the cosmetic or dermatological composition containing it, formulated in its complete culture medium. origin as defined below, which complete culture medium is the medium in which the bacteria were cultured until after the microbial growth phase having led to the use of nutrient substrates initially present in the culture medium. Within the meaning of the present invention, the expression "complete fermentation medium" is intended to mean a medium resulting from the culture process that has been used for the growth and cell lysis of the microorganism, said medium having also undergone no additional manipulation. to separate and / or remove all or part of its non-aqueous components. For the purposes of the present invention, the term "preventing" means completely eliminating or partially reducing the risk of manifestation of a given phenomenon, that is to say in the present invention the manifestation of dandruff states. scalp.

The partial decrease implies that the risk remains but to a lesser degree than before the implementation of the invention. A composition containing the active agent according to the invention can be administered topically.

Application of an extract of non-photosynthetic non-fruiting filamentous bacteria to the image of bacteria of the genus Vitreoscilla sp. (Especially species Vitreoscilla fil formis) as active to modulate and preferably inhibit the adhesion and / or proliferation of pathogenic microorganisms on the skin and scalp has already been proposed in document FR 2 879 452.

However, the extract considered in this document consists of either the supernatant of the fermentation medium of said bacterium, or the biomass obtained after culturing said bacteria, or the envelopes or fractions of envelopes or extracts obtained following a complementary treatment of the biomass. Consequently, all the above-mentioned extracts are obtained only after a complementary operation such as, for example, filtration or centrifugation, imposed on the fermentation medium in order to separate the extract under consideration from the other constituents of this fermentation medium. The active agent according to the invention is therefore totally distinct from the extract under consideration and described in the document FR 2 879 452. In contrast to the extract described in FR 2 879 452, the active agent according to the invention is constituted of all the components present in the fermentation medium. The surprising effect of the use according to the invention therefore results from the use, as an active ingredient, for the first time by the inventors of a microorganism lysate in its complete fermentation medium. Indeed, and as is apparent from the examples below, the inventors have found that the active ingredient according to the invention has a regulating activity of the microflora of the scalp, greater than that observed for a biomass type extract of the same bacterium. . In particular, the inventors have observed that at the end of a treatment of the invention, the balance of the microflora and the barrier properties of the scalp were reinforced. On the other hand, it has been found by the inventors a beneficial effect of the active agent according to the invention on the stimulation of antimicrobial peptides. As shown by the examples below, an active agent in accordance with the invention obviously stimulates the expression of the Pdef 4 gene. This effect has notably been verified by qRT-PCR from a reconstructed Realskin® skin model. Without being bound by what follows, this efficiency gain could be the result of a synergistic effect between constituents of the bacterium, usually separated from each other, for example its water-soluble metabolites, generated during its proliferation in its fermentation medium and conventionally present in the aqueous supernatant and its components such as envelopes or non-water-soluble cell shell fractions constituting all or part of the biomass of its culture medium or even its isolated lysate. It has thus been shown in the examples that a cosmetic composition comprising an active ingredient according to the invention (a lysate of bacteria belonging to the genus Vitreoscilla sp., In a complete fermentation medium) has equal reduction properties of the scalp dandruff conditions. or even superior to known antidandruff active agents, such as octopyrox or zinc pyrythione (ZnPT). In particular, it has been shown in the examples that a cosmetic formulation comprising an active ingredient of the invention is active against dandruff scalp conditions at doses much lower than the conventional doses of anti-dandruff active ingredients required to obtain the same effects on the dandruff of the scalp. It has also been shown that an active agent according to the invention also has scalp itching properties caused by the dandruff states. Unexpectedly, the active ingredient in question according to the invention seems on the other hand devoid of inhibitory activity with regard to the various strains of malassezia sp studied: globosa and furfur, as shown in the examples. According to another embodiment, the present invention relates to a cosmetic method for preventing and / or treating dandruff conditions of the scalp, including dandruff scalp conditions associated with a prevalence of pathogenic microorganisms on the scalp and / or an imbalance of ecoflora of the scalp in an individual, comprising at least one step of administering to said individual an effective amount of at least one bacterial lysate (s) in a complete fermentation medium, said lysate in its environment; complete fermentation being capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp. For the purposes of the present invention, the term "effective amount" means a sufficient and necessary amount of the compound in question to obtain the expected effect. Such an amount can be determined by any method known to those skilled in the art, for example by means of preliminary experimental tests. According to another of its aspects, the present invention relates to a rinsed or non-rinsed cosmetic and / or dermatological composition containing in a physiologically acceptable medium at least one lysate of microorganism (s) in a complete fermentation medium, said lysate in its medium complete fermentation being capable of inducing or stimulating the production of at least one antimicrobial peptide in the scalp. Within the meaning of the present invention, the strains of microorganisms concerned by the invention and whose use in the form of a lysate in the complete fermentation medium is sought, are in particular strains of "probiotic microorganism". These microorganisms that are suitable for the invention may be chosen in particular from ascomycetes such as Saccharomyces, Yarrowia, Kluyveromyces, Torulaspora, Schizosaccharomyces, Debaromyces, Candida, Pichia, Aspergillus and Penicillium, bacteria of the genus Bifidobacterium, Bacteroides, Fusobacterium, Melissococcus, Propionibacterium. , Enterococcus, Lactococcus, Staphylococcus, Streptococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus and mixtures thereof. Particularly suitable as probiotic ascomycetes for the present invention are Yarrowia lipolitica and Kluyveromyces lattis, as well as Saccharomyces cereviseae, Torulaspora spp., Schizosaccharamyces pombe, Candida spp. and Pichia spp. Specific examples of probiotic batteries are Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus casei subsp. Casei, Lactobacillus casei Shirota, Lactobacillus paracasei, Lactobacillus curvatus, Lactobacillus delbruck, subsp. Lactis, Lactobacillus gasseri, Lactobacillus johnsonum, Lactobacillus reuteri, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus sake, Lactococcus lattis, Streptococcus thermophilus, Staphylococcus carnosus, and Staphylococcus xylosus and mixtures thereof. More particularly, it is probiotic microorganisms from the group of lactic acid bacteria, such as Lactobacillus and / or Bifidobacterium. As an illustration of these lactic acid bacteria, mention may be made more particularly of Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei or in the genus Bifidobacterium species, among which: Bifidobacterium longuet, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium animes , Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium adolescentis or Bifidobacterium pseudocatenulatum, and mixtures thereof. In strains of probiotic bacteria, the most suitable species are Lactobacillus johnsonii, Bifidobacterium adolescentis, Bifidobacterium longum respectively deposited according to the Budapest Treaty with the Pasteur Institute (28 rue du Docteir Roux, F-75024 Paris cedex 15) under the designations CNCM I-1225, CNCM I-2168 and CNCM I-2170, and the genus Bifidobacterium lactis (Bb 12) (ATCC27536). The strain of Bifidobacterium lactis (ATCC27536) can be obtained from Hansen (Hansen AIS Chr., 10-12 Boege Alle, PO Box 407, DK-2970 Hoersholm, Denmark).

Complete fermentation medium As stated above, the term "complete fermentation medium" means a fermentation medium or culture having the same composition, at least in terms of non-aqueous components or in its entirety, as the fermentation medium in which the microorganism dedicated to forming the parallel required lysate according to the invention was successively fermented and lyzed. In other words, this medium has not undergone any other manipulation aimed at separating and / or eliminating all or part of its non-aqueous constituents. More particularly, the active agent according to the invention consists of the lysate of microorganisms and all or part, in terms of quantity, of the culture medium having served for the fermentation of said bacterium and in which it has proceeded consecutively to its cell lysis (ie complete fermentation medium). In view of the foregoing, it appears that the active compound according to the invention from the lysate and the so-called complete fermentation medium contains the cytoplasmic and cytosolic fractions, the cell wall fragments and the metabolites formed and / or released during cell lysis of said microorganism, the entirety of biological entities capable of being generated and released spontaneously by the bacterium during its fermentation process and therefore already present in the fermentation medium before cell lysis thereof. Consequently, an active agent according to the invention, that is to say formed from a lysate of bacteria in a complete fermentation medium according to the invention, is clearly different from the supernatant of a fermentation medium of said bacterium, as this is illustrated in the examples with a bacterium belonging to the genus Vitreoscilla sp. (Especially species: Vitreoscilla filiformis). Indeed, the active agent according to the invention, as opposed to the supernatant, contains cellular fragments of said bacterium represented by the lysate.

An active agent according to the invention, that is to say formed of a bacterial lysate belonging to the genus Vitreoscilla sp. (In particular species: Vitreoscilla filiformis) in a complete fermentation medium according to the invention, capable of inducing or stimulating the production of at least one antimicrobial peptide, is also different from the biomass or fraction of biomass, or even from a lysate or fraction of lysate isolated from a fermentation medium of said bacterium, as illustrated in the examples with a bacterium belonging to the genus Vitreoscilla sp. (Especially species: Vitreoscilla filiformis). In fact, the active ingredient according to the invention, as opposed to this biomass, or fraction of biomass, this lysate or lysate fraction contains a significant amount of water-soluble metabolites released naturally into the culture medium during the proliferation of said bacterium. For the purposes of the present invention, the expression "non-aqueous constituents" means that water, which is a major constituent of conventional fermentation media, is not one of the constituents that must remain as such, that is, say equal amount, in the complete culture medium according to the invention.

Thus, the term "complete medium" also extends to a form of complete medium which is said to be concentrated in view of the fact that it is obtained after partial evaporation of the water constituting a fermentation medium in which consecutively carried out the culture of the corresponding microorganism and the cell lysis thereof. Of course, this evaporation is conducted under operating conditions adjusted so as not to alter the integrity of the non-aqueous constituents forming this complete medium.

Fermentation medium composition By definition, a fermentation medium or culture is a support that allows the culture and therefore, as appropriate, the development of cells, bacteria, yeasts .. In principle, the cells found in this medium the essential components for their multiplication in large numbers quickly, but also sometimes elements that will favor the development of a specific bacterial genus or a particular family, in this case a bacterium whose lysate in its complete fermentation medium is capable of inducing or stimulating the production of at least one antimicrobial peptide, including a bacterium belonging to the genus Vitreoscilla sp. (Notably, species: Vitreoscilla filiformis).

Its composition must therefore meet the nutritional requirements of the microorganism considered and necessary for the proliferation thereof. More precisely, the composition of this culture medium must: - cover the needs for mineral ions, growth factors, provide the source of carbon and energy; - have a pH close to the optimal pH and; have an optimal ionic strength (the medium can be isotonic but it is not obligatory).

Thus, the composition of a fermentation medium suitable for the invention comprises at least one; a source of carbon and energy, generally represented by a sugar and advantageously glucose, a source of potassium and phosphorus, for example image of K 2 HPO 4, a source of nitrogen and sulfur which can be represented by compound (NH4) 2SO4; a source of magnesium such as, for example, MgCl 2, a source of calcium such as for example CaCl 2, a source of iron and more particularly iron citrate, the role of the citrate being to keep the iron in solution, source of trace elements chosen in particular from the salts of Cu, Zn, Co, Ni, B, - a source of water, generally sterile, essential for all life, - a pH buffer that can be represented by KH2PO4. In the absence of one of these components, the bacteria do not develop, because they can not by themselves make up for its absence. As an illustration of a fermentation medium suitable for the development of a microorganism according to the invention, mention may in particular be made of the medium shown in Example 1 below. An effective amount of the microorganism considered according to the invention is introduced therein and the whole is placed under conditions conducive to its proliferation. To obtain a lysate of bacteria concerned by the invention, for example a lysate of a bacterium belonging to the genus Vitreoscilla sp. in a complete fermentation medium, according to a process comprising a step of culturing said bacteria, one skilled in the art can refer in particular to Example 1. Advantageously, to obtain an active agent according to the invention, that is to say say a lysate of bacteria concerned by the invention, for example a lysate of bacteria belonging to the genus Vitreoscilla sp., in a complete fermentation medium, the biomass (bacterial cells after the growth phase, present in the medium in which they have cultivated) is frozen, for example at a temperature of -20 ° C, and is then sterilized, preferably by heat, in particular by subjecting the previously frozen biomass to a heating step at a temperature above 100 ° C. By way of illustration, the step of sterilizing the biomass may be carried out by autoclaving, for example at a temperature of 121 ° C. As is apparent from the foregoing, at the end of this culture of said microorganism that is transformed into the lysate state directly within the fermentation medium used for its culture.

Lysis of bacteria to obtain an active agent according to the invention A lysate commonly refers to a material obtained at the end of the destruction or dissolution of biological cells by a phenomenon called cell lysis thus causing the release of the intracellular biological constituents and naturally contained in the biological cells under consideration. For the purposes of the present invention, the term lysate refers to the entirety of the lysate obtained by lysis of the microorganism in question, namely a bacterium whose lysate in its complete fermentation medium is capable of inducing or stimulating the production of minus an antimicrobial peptide, including a bacterium belonging to the genus Vitreoscilla sp. (in particular species: Vitreoscilla filiformis). The lysate used is therefore formed of all of its biological constituents, intracellular in particular its metabolites and the constituents of the cell walls and membranes generated during its cell lysis. For the purposes of the invention, the term "metabolite" refers to any substance derived from the metabolism of the microorganism in question according to the invention. This cell lysis can be accomplished by a variety of technologies, such as, for example, osmotic shock, thermal shock, ultrasound, or mechanical stress centrifugation. More particularly, this lysate can be obtained according to the technology described in US Pat. No. 4,464,362, and in particular according to the following protocol. Ultrasonic disintegration of the fermentation medium used in the culture of the microorganism considered and thus containing said microorganism to release cytoplasmic and cytosolic fractions, cell wall fragments and metabolic products is carried out. of this microorganism. All these components are then preserved in their natural distribution in a stabilized form in the so-called complete fermentation medium. Accordingly, the active agent according to the invention can be obtained via a process consisting of the cultivation of at least one bacterium belonging to the genus Vitreoscilla sp. (Especially species: Vitreoscilla filiformis) on a fermentation medium under conditions conducive to the proliferation of said bacterium, and cell lysis of said bacteria in said fermentation medium.

Bacteria belonging to the genus Vitreoscilla sp. (In particular species: Vitreoscilla filiformis) As stated above, the microorganism considered according to the invention in the lysate state is a non-synthetic filamentous bacterium as defined according to the Bergey's Manual of Systematic Bacteriology classification (Vol. sections 22 and 23, 9th edition, 1989), and belonging to the genus Vitreoscilla sp. (Especially species: Vitreoscilla filiformis). As can be seen from the examples presented below, the inventors have unexpectedly discovered that such a bacterium used in the form of its lysate formulated in a complete fermentation medium within the meaning of the present invention has superior properties. term of efficiency to those of the biomass obtained from the same fermentation medium. More particularly, it is a bacterium belonging to the genus Beggiatoa, Vitreoscilla, Flexithrix or Leucothrix. Examples of suitable bacteria that may be mentioned include Vitreoscilla filiformis (ATCC 15551). According to a preferred variant of the invention, it is the bacterium Vitreoscilla filiformis.

Applications As previously indicated, the active agent according to the present invention is particularly interesting with regard to its effectiveness with regard to certain cosmetic disorders related to dandruff conditions of the scalp, associated with a prevalence of pathogenic microorganisms on the scalp and / or an imbalance of the ecoflora of the scalp. An active agent according to the invention is advantageously used for the purpose of treating and / or preventing the dandruff states and the associated scalp disorders. Thus, a scalp with excessive dryness or excessive secretion of sebum may show a dandruff condition, which, as the case may be, may be characterized by the presence of dry or greasy dandruff, or even pruritus and / or inflammation of the skin. 'epidermis. The dry dandruff states translate xerosis of the scalp, possibly associated with an excessively rapid renewal of its stratum corneum. Dry dandruff is usually small in size, white or gray, and spreads over the scalp and clothing that creates an unsightly visual effect. As for the fatty dandruff states, they appear in one of the sub-pathological forms of seborrheic dermatitis.

In the dandruff states of the scalp, the skin barrier is unbalanced, its integrity and hydration are altered and its ecoflora disturbed. The skin of the scalp is irritated and itchy, fragile, less hydrated and susceptible to infections. As can be seen from the examples below, the present invention advantageously makes it possible to have a new active that is particularly effective with regard to the dandruff states. Advantageously, an active agent according to the invention is capable of reducing the risk of occurrence of side effects. The active agent according to the invention still advantageously makes it possible to restore a healthy scalp in perfect homeostasis and to restore a balanced ecoflor. Consequently, the cosmetic uses, cosmetic processes and compositions according to the invention prove to be particularly effective. to improve hygiene and / or care of the scalp, and in particular to prevent and / or treat dandruff conditions, whether dry or greasy, of the scalp, for preventing and / or treating disorders, particularly aesthetic, scalp associated with excessive excretion and / or secretion of sebum, - to impart a feeling of well-being to the scalp, 20 to improve the comfort of the scalp, to improve and / or restore the defenses Endogenous antimicrobial scalp, to preserve and / or enhance the integrity of barrier functions of the scalp, 25 to restore a balanced ecoflora of leather hairy, to prevent and / or treat pruritus and itching of seborrheic origin associated with the dandruff condition of the scalp.

In general, the present invention is directed to a composition comprising, as a sole anti-dandruff active agent, a lysate of bacterium (s) belonging to the genus Vitreoscilla sp. In a complete fermentation medium However, the invention also provides a composition comprising, besides a lysate according to the invention in a complete fermentation medium, at least one additional dandruff active agent and / or at least one distinct cosmetic active.

Additional cosmetic active A method according to the invention may comprise, in addition to the administration of an active agent according to the invention in a form or not associated with an anti-dandruff active agent, the administration of at least one third cosmetic active agent. Advantageously, such an additional cosmetic active may be intended to exert a cosmetic effect, care or hygiene on the scalp, or even be intended to strengthen the cutaneous barrier of the scalp. It may especially be a probiotic microorganism, and / or one of its fractions, and / or one of its metabolites, distinct from said bacterium considered in the active according to the invention and / or the complete culture of this bacterium. microorganism. Probiotic microorganism For the purposes of the present invention, the term "probiotic microorganism" means a living microorganism which, when consumed in an adequate quantity, has a positive effect on the health of its host ("Joint FAO / WHO Expert Consultation" In particular, the present invention can improve the intestinal microbial equilibrium. According to one embodiment, a probiotic microorganism that is suitable for the invention may be chosen from Lactobacillus sp., Bifidobacterium sp., Cocci, yeasts, sporulated bacteria, and mixtures thereof. According to one embodiment, a microorganism that is suitable for the invention is preferentially chosen from lactic acid bacteria: which produce by fermentation sugar of lactic acid. According to their morphology they are divided into two groups: Lactobacillus species: Lactobacillus acidophilus, amylovorus, casei, rhamnosus, brevis, crispatus, delbrueckii (subsp bulgaricus, lactis), fermentum, helveticus, 29.73699 gallinarum, gasseri, johnsonii, plantarum, reuteri salivarius, alimentarius, curvatus, casei subsp. casei, sake, and Cocci: Enterococcus (faecalis, faecium), Lactococcus lactis (subsp lactis or cremoris), Leuconostoc mesenteroides subsp dextranicum, Pediococcus acidilactici, Sporolactobacillus inulinus, Streptococcus salvarius subsp. thermophilus, Streptococcus thermophilus, Staphylococcus carnosus, Staphylococcus xylosus, - bifidobacteria or Bifidobacterium species: Bifidobacterium adolescentis, animalis, bifidum, breve, lactis, longet, infantis, pseudocatenulatum, - yeasts: Saccharomyces (cerevisiae or boulardii), - the others Spore-forming Bacillus bacteria (Cereus var toyo or subtilis), Bacillus coagulans, Bacillus licheniformis, Escherichia coli strain nissle, Propionibacterium freudenreichii, and mixtures thereof. A microorganism that is suitable for the invention may be chosen especially from ascomycetes such as Saccharomyces, Yarrowia, Kluyveromyces, Torulaspora, Schizosaccharomyces pombe, Debaromyces, Candida, Pichia, Aspergillus and Penicillium, bacteria of the genus Bifidobacterium, Bacteroides, Fusobacterium, Melissococcus, Propionibacterium. , Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus, and mixtures thereof. As another example of probiotic microorganisms that are suitable for the invention, mention may be made of Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium lactis, Bifidobacterium longet, Bifidobacterium infantis, Bifidobacterium pseudocatenulatum and Lactobacillus acidophilus NCFB 1748; Lactobacillus amylovorus, Lactobacillus casei (Shirota), Lactobacillus rhamnosus strain GG, Lactobacillus brevis, Lactobacillus crispatus, bulgaricus, Lactobacillus delbrueckii subsp., Lactis, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus johnsonii CNCM I-1225, Lactobacillus plantarum , Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus alimentarius, Lactobacillus curvatus, Lactobacillus casei subsp. Casein, Lactobacillus sake, Lactococcus lactis, Enterococcus faecalis, Enterococcus faecium, Lactococcus lactis subsp lactis, Lactococcus lactis subsp cremoris, Leuconostoc. 2973699 Mesenteroides subsp dextranicum, Pediococcus acidilactici, Sporolactobacillus inulinus, Streptococcus salvarius subsp. thermophilus, Streptococcus thermophilus, Staphylococcus carnosus, Staphylococcus xylosus, Saccharomyces cerevisiae, Saccharomyces boulardum, Bacillus cereus var toyo, Bacillus cereus var subtilis, Bacillus coagulans, Bacillus licheniformis, Escherichia coli strain nissle, Propionibacterium freudenreichii, and mixtures thereof. More particularly, it may be a probiotic microorganism selected from Lactobacillus sp., Sporolactobacillus sp., Enterococcus sp., Lactococcus sp., Bacillus sp., Streptococcus sp., Pediococcus sp., Leuconostoc sp. or Bififobacterium sp., and in particular selected from Lactobacillus sp. and Bifidobacterium sp., and mixtures thereof. As an illustration of these probiotic microorganisms, mention may be made more particularly of Lactobacillus johnsonum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Bifidobacterium animes, Bifidobacterium lactis, Bifidobacterium infantis and Bifidobacterium adolescentis. , Bifidobacterium pseudocatenulatum, and mixtures thereof. The most suitable species are Lactobacillus johnsonum, Lactobacillus paracasei, Bifidobacterium adolescentis, Bifidobacterium longum respectively deposited according to the Budapest Treaty with the Institut Pasteur (28 rue du 20 Dr. Roux, F-75024 Paris cedex 15) on 30/06/92 , 12/01/99, 15/04/99 and 15/04/99 under the following designations CNCM I-1225, CNCM I-2116, CNCM I-2168 and CNCM I-2170, and the genus Bifidobacterium lactis (Bb 12) (ATCC27536) or Bifidobacterium longum (BB536). The strain of Bifidobacterium lactis (ATCC27536) can be obtained from Hansen (Hansen A / S Chr., 10-12 Boege Alle, PO Box 407, DK-2970 Hoersholm, Denmark). Advantageously, a microorganism that is suitable for the invention, as an additional active ingredient, may be a probiotic microorganism with lactic acid. According to a preferred embodiment, a probiotic microorganism that is suitable for the invention may in particular be a microorganism of the genus Lactobacillus sp.

Preferably, a microorganism of the genus Lactobacillus sp. suitable for the invention may be chosen from the species Lactobacillus johnson, Lactobacillus reuteri, Lactobacillus paracasei, Lactobacillus casei, and mixtures thereof. According to a preferred embodiment, a microorganism suitable for the invention may be a Lactobacillus paracasei. A microorganism that is suitable for the invention may in particular be the strain Lactobacillus paracasei ST11 deposited according to the Budapest Treaty at the Institut Pasteur (28 rue du Docteur Roux, F-75024 Paris cedex 15) on 12/01/99 under the designation CNCM I-2116, and / or a fraction thereof and / or a metabolite thereof. According to another preferred embodiment, a probiotic microorganism that is suitable for the invention may in particular be a microorganism of the genus Bifidbacterium sp., And in particular Bifidobacterium longum, in particular Bifidobacterium longum (BB536). Such a microorganism may be formulated in a composition in an amount of at least 0.0001%, expressed in dry weight, in particular in the range of 0.0001 to 20% and more particularly in the proportion of 0.001 to 15% by weight, in in particular from 0.01 to 10% by weight, and especially from 0.1% to 2% by weight relative to the total weight of the composition containing it.

Galenic Formulation A composition containing the active ingredient according to the invention may be administered orally or topically. A composition according to the invention advantageously comprises a quantity of bacterial lysate (s) belonging to the genus Vitreoscilla sp. in a complete fermentation medium ranging from 0.001% to 10% by weight, based on the total weight of dry extract of said composition. In certain embodiments, a composition according to the invention advantageously comprises a quantity of bacterial lysate (s) belonging to the genus Vitreoscilla sp. in a complete fermentation medium ranging from 0.01% to 5% by weight, relative to the total weight of dry extract of said composition. In other embodiments, a composition according to the invention advantageously comprises a quantity of bacterial lysate (s) belonging to the genus Vitreoscilla sp. in a complete fermentation medium ranging from 0.1% to 1% by weight, relative to the total weight of dry extract of said composition. It can therefore be in any galenic form normally available for the selected mode of administration.

The support may be of a different nature depending on the type of composition considered. As regards more particularly the compositions intended for external topical administration, they may be aqueous, hydroalcoholic or oily solutions, solutions or dispersions of the lotion or serum type, emulsions of liquid or semi-liquid consistency. of the milk type, obtained by dispersion of a fatty phase in an aqueous phase (O / W) or conversely (W / O), or suspensions or emulsions, of soft, semi-solid or solid consistency, of the cream type, of aqueous or anhydrous gel, microemulsions, microcapsules, microparticles, or vesicular dispersions of ionic and / or nonionic type. These compositions are prepared according to the usual methods.

These compositions may include creams for cleaning, protection, treatment or care of lotions, gels or foams for the care of the scalp, such as cleaning lotions or disinfection of the scalp. They can be used for the scalp in the form of solutions, creams, gels, emulsions, foams or in the form of aerosol compositions also containing a propellant under pressure. A composition topically according to the invention may advantageously be formulated in any dosage form suitable for hair care, especially in the form of a hair lotion, a shampoo, especially anti-dandruff, a conditioner, a detangler , a cream or a hair gel, a hairspray, a styling lotion, a treatment lotion, a dyeing composition (in particular oxidation) optionally in the form of a shampoo dye, a restructuring hair lotion, a permanent composition, a lotion or a fall protection gel, a pest control shampoo, or a treatment shampoo, in particular anti-seborrhoeic, of a care product scalp especially anti-irritant, anti-aging, restructuring, or 'activator of the blood circulation. When a composition of the invention is an emulsion, the proportion of the fatty phase can range from 5 to 80% by weight, and preferably from 10 to 50% by weight relative to the total weight of the composition. The oils, emulsifiers and coemulsifiers used in the composition in emulsion form are chosen from those conventionally used in the cosmetics and / or dermatological field. The emulsifier and the coemulsifier may be present in the composition in a proportion ranging from 0.3 to 30% by weight, and preferably from 0.5 to 20% by weight relative to the total weight of the composition. When the composition of the invention is a solution or an oily gel, the fatty phase may represent more than 90% of the total weight of the composition. In known manner, the dosage forms dedicated to topical administration may also contain adjuvants which are usual in the cosmetic, pharmaceutical and / or dermatological field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, odor absorbers and dyestuffs. The amounts of these various adjuvants are those conventionally used in the field under consideration, and for example from 0.01 to 20% of the total weight of the composition. These adjuvants, depending on their nature, can be introduced into the fatty phase and / or into the aqueous phase. Fats which can be used in the invention include mineral oils such as, for example, hydrogenated polyisobutene and petrolatum oil, vegetable oils such as, for example, a liquid fraction of shea butter, sunflower oil and apricot kernels, animal oils such as perhydrosqualene, synthetic oils including Purcellin oil, isopropyl myristate and ethyl hexyl palmitate, unsaturated fatty acids and fluorinated oils, for example perfluoropolyethers. It is also possible to use fatty alcohols, fatty acids such as, for example, stearic acid and, for example, waxes such as paraffin wax, carnauba wax and beeswax. It is also possible to use silicone compounds such as silicone oils and, for example, cyclomethicone and dimethicone, waxes, resins and silicone gums. As emulsifiers that can be used in the invention, mention may be made, for example, of glycerol stearate, polysorbate 60, the cetylstearyl alcohol / cetylstearyl alcohol mixture oxyethylenated with 33 moles of ethylene oxide sold under the name Sinnowax AOC by the company HENKEL, the mixture of PEG-6 / PEG-32 / glycol stearate sold under the name Tefose® 63 by Gattefosse, PPG-3 myristyl ether, silicone emulsifiers such as cetyldimethicone copolyol and sorbitan mono- or tristearate, PEG-40 stearate, oxyethylenated sorbitan monostearate (200E). As solvents that can be used in the invention, mention may be made of lower alcohols, especially ethanol and isopropanol, and propylene glycol. The composition of the invention may also advantageously contain a thermal and / or mineral water, in particular chosen from Vittel water, the waters of the Vichy basin and Roche Posay water. As hydrophilic gelling agents, mention may be made of carboxylic polymers such as carbomer, acrylic copolymers such as copolymers of acrylates / alkyl acrylates, polyacrylamides and in particular the mixture of polyacrylamide, C13-14-Isoparaffin and Laureth-7 sold under the Sepigel 305® by the company SEPPIC, polysaccharides such as cellulose derivatives such as hydroxyalkylcelluloses and in particular hydroxypropylcellulose and hydroxyethylcellulose, natural gums such as guar, carob and xanthan and clays. As lipophilic gelling agents, mention may be made of modified clays such as bentones, metal salts of fatty acids such as aluminum stearates and hydrophobic silica, or else ethylcellulose and polyethylene. Cosmetic treatment method As indicated above, a process according to the invention may be carried out topically, in particular by application to the scalp of at least one bacterial lysate concerned by the invention, including a bacterium belonging to the genus Vitreoscilla sp. (In particular, species: Vitreoscilla filiformis) in a complete fermentation medium as an active ingredient for preventing and / or treating body hygiene disorders, particularly associated with a proliferation of pathogenic microorganisms on the scalp and / or an imbalance of the body. ecoflore of the scalp, and more particularly of a cosmetic composition as defined above. Advantageously, a method of the invention topically can comprise the application of a composition according to the invention, for example in the form of cleaning creams, lotions, or treatment or care gels for the scalp, gels or mousses for the care of the scalp, such as cleaning or disinfecting lotions or shampoos. According to an alternative embodiment, the method of the invention may comprise the topical application to the scalp of at least one active ingredient according to the invention, for example in the form of shampoo, gels, serums or lotions. A topical cosmetic method according to the invention may be used daily for example, for example for example a single administration per day or a twice daily administration, for example once in the morning and once the evening. A topical cosmetic process according to the invention may be carried out over a period of time varying from one week to several weeks, or even several months, this period being able to be repeated after periods of non-treatment, for several months, even several years. By way of example, topical administration of a compound according to the invention may be repeated, for example, 2 to 3 times a day, or more, and generally over an extended period of at least 4 weeks. or 4 to 15 weeks, possibly with one or more periods of interruption. In the following description and examples, unless otherwise indicated, percentages are percentages by weight and ranges of values in the form "between: .. and ..." include the specified lower and upper bounds. The following examples are presented by way of illustration and not limitation of the field of the invention.

EXAMPLES EXAMPLE 1 Preparation of an Active Ingredient According to the Invention The production of the complete fermentation medium is obtained by means of a culture of the Vitreoscilla filiformis strain, in its complete culture medium. The initial culture medium for obtaining the complete fermentation medium is of composition described in Table 1 below. Table 1 Chemical name [c] Autolytic yeast extract 4 g / 1 Soya papainic peptone F 3 g / 1 Glucose - Roferose 3 g / 1 KH2PO4 0.088 g / 1 CaCl2 0.050 g / 1 CuSO4, 5 H2O 60 gg / 1 MnSO4.1 H2O 152 μg / 1 KI 20 μg / l ZnSO4, 7H20 200 μg / 1 AlC13, 6H2O 100 μg / 1 osmosis water qs To obtain a cosmetic active agent according to the invention, that is to say in In this example, a lysate of Vitreoscilla filiformis bacteria in a complete fermentation medium was carried out as described below. The Vitreoscilla filiformis strain was obtained from ATCC (strain 15551). This strain is cultured in a particular culture medium 2BHG2, the composition of which is given above. Biomass is obtained by continuous cultivation in a 3000 liter efficient bioreactor. A growth rate of about 70% of the pmax (μ = 0.12 H-1) is recorded during the continuous production phase. During this step, we control pH (7.00), temperature (26 ° C) and dissolved oxygen (0/5%). The extraction and separation of the cells is obtained by centrifugation (10,000 g / 20 min). The biomass is frozen at 20 ° C. and then the biomass is packaged in bags (rupture of sterility) and is stabilized by sterilization at 121 ° C. for 30 minutes. The biomass is then called Vfe. The biomass specification analyzes are as follows: - Fixed residue at 105 ° C (g / 100g): 4.0 to 4.5% - Total nitrogen content in MA: 10.0 to 14.0% - Microbiology: 0 germ / g - Content in 3-hydroxybutyric acid: 2 to 10 g / l (<10 g / l) pH of the solution in state: 4 to 5. The fermentation medium is the complete culture, obtained during the continuous fermentation. The glucose of the starting medium was consumed by the g-organisms (μ-controlled by the carbon source), as well as various elements of the peptones and yeast extract brought to the start. The fermentation medium currently being tested is taken directly from the fermentor and then undergoes the sterilization scale. MF, which is the complete non-concentrated culture (0.7 to 0.9% DM) is autoclaved (30 min 121 ° C), as well as the lysate (4.0 to 4.5% DM).

In the examples which follow, the active agent according to the invention (the lysate of Vitreoscilla filiformis bacteria in a complete fermentation medium, also called "MF") as well as a comparative active agent consisting of the isolated bacterial lysate in the absence of the medium. For complete fermentation, clinical head tests were performed to check their anti-dandruff activity compared to a standard anti-dandruff shampoo formulation. In particular, two reference anti-dandruff shampoo formulations were tested, respectively a first reference shampoo formulation comprising octopyrox and a second reference shampoo formulation comprising ZnPT.

EXAMPLE 2 In Example 2, the effects on the dandruff conditions of the scalp (i) of a shampoo formulation comprising an anti-dandruff active agent according to the invention (lysate of Vitreoscilla filiformis bacteria in a fermentation medium were compared. complete, prepared according to Example 1) and (ii) a standard anti-dandruff shampoo formulation, free of the anti-dandruff active according to the invention, comprising a conventional anti-dandruff active (ZnPT). The lysate of bacteria in a complete fermentation medium (anti-dandruff active according to the invention) as obtained in Example 1 is therefore used as an anti-dandruff active in a shampoo formulation whose basic composition is described in Table 2 below. Several clinical studies have been performed, - a clinical study 1, corresponding to Figure 1, - a clinical study 2, corresponding to Figure 2, - a clinical study 3, corresponding to Figures 3 to I1.

Table 2 Ingredients Concentration (% by weight based on total weight) Chemical name INCI name (US) INCI name (LIE) Vitreoscilla 2 filimormis lysate in their complete medium (complete culture at 160g / 1) Sodium laureth Sodium laureth Lauryl ether sulfate laureth 20.1 sodium (2.2 0E) sulphate sulfate aqueous solution Cocoyl amidopropyl Cocamidopropyl Cocamidopropyl 6.3 betaine solution betaine aqueous betaine Water QSP The shampoo formulation, used in Clinic 3, Table 2 comprises 2% by weight of l anti-dandruff active according to the invention (lysate of bacteria in a complete fermentation medium, at a concentration of 160 g / l) relative to the total weight of the composition, that is to say 0.32% by weight of said anti-dandruff active, based on the total weight of dry extract of said composition. Equivalently, the shampoo formulation used in Clinic 3 comprises 1% by weight of the anti-dandruff active according to the invention (lysate of bacteria in a complete fermentation medium, at a concentration of 160 g / l) relative to total weight of the composition, that is to say 0.16% by weight of said anti-dandruff active agent, relative to the total weight of dry extract of said composition. Equivalently, the shampoo formulation used in Clinic 1 comprises 2.5% and 5% by weight of the bacterial lysate active (bacterial lysate free of the complete fermentation medium) relative to the total weight of the composition; that is to say respectively 0.1075% and 0.215% by weight of said active agent, relative to the total weight of dry extract of said composition. Equivalently, the shampoo formulation used in Clinic 2 comprises 20% by weight of the anti-dandruff active agent according to the invention (lysate of bacteria in a complete fermentation medium, at a concentration of 7.5 g / l). relative to the total weight of the composition, that is to say 0.14% by weight of said anti-dandruff active agent, relative to the total weight of dry extract of said composition. It is specified that the amount in% by weight of the lysate in its complete fermentation medium varies from 0.1% to 0.5% o in dry extract.

By way of comparison, the reference shampoo formulation described in Table 3 below, which comprises the zinc pyrythione anti-dandruff active agent (ZnPT) present at 2% by weight, relative to the total weight of the composition, was tested. , that is to say 1% of dry weight by weight zinc pyrythione, relative to the total weight of dry extract of said composition. Table 3 Ingredients Concentration (% by weight based on total weight) Chemical Name INCI Name (US) INCI Name (EU) Zy Pyrythione Zinc pyrythione Zinc pyrythione 2.083, which in dispersion is equivalent to 1% aqueous by weight Lauryl ether sulphate Sodium laureth Sodium laureth 20.1 sodium (2.2 0E) sulphate sulphate in aqueous solution Cocoyl amidopropyl Cocamidopropyl Cocamidopropyl 6.3 betaine in solution betaine aqueous betaine Water QSP Protocol The treatment consists in applying topically the formula tested during four weeks.

Clinical study 1 This study was initiated to compare (i) a shampoo comprising an isolated bacterial lysate, free of complete fermentation medium, used at 2.5% by weight (corresponding to 0.1075% of dry extract), (ü) a shampoo comprising an isolated bacterial lysate, free of complete fermentation medium, used at 5% by weight (corresponding to 0.215% solids), and (iii) the standard anti-dandruff shampoo formula. This study was conducted in 60 adult male subjects between the ages of 18 and 60 who were identified following a clinical assessment of their moderate to severe dermal condition with scores greater than or equal to 5 (on a scale of 0 to 9) in the presence of adherent scales on at least two quarters of the head, and their erythema of the scalp. The 60 subjects were divided into 3 parallel groups of 20 subjects, with each group being treated with only one of the two compositions + a control group

Clinical Study 2 This study was initiated to compare (i) a shampoo comprising the active ingredient of the invention at 20% by weight (corresponding to 0.14% solids), and (ii) the shampoo formula reference. The active ingredient of the invention is in a form at 7.5 g / l. This study was conducted on 42 adult male subjects aged between 18 and 60 years who were identified following a clinical evaluation of their moderate to severe dermal condition with scores greater than or equal to 5 (on a scale of 0 to 9) in the presence of adherent scales on at least two quarters of the head, and their erythema of the scalp. The 42 subjects were divided into 2 parallel groups of 21 subjects, with one group being treated with a composition presenting the active ingredient of the invention + a control group with the reference anti-dandruff active.

Clinical Study 3 This study was initiated to compare (i) a shampoo comprising the active ingredient of the invention with 0.16% dry extract, (ii) a shampoo comprising the active ingredient of the invention at 0.32% extract sec and (iii) the reference shampoo formula, comprising zinc pyrythione at 1% dry extract. The active ingredient of the invention is in a form at 160 g / l. This study was conducted in 67 adult male subjects between the ages of 18 and 60 who were identified following a clinical assessment of their moderate to severe dermal condition with scores greater than or equal to 5 (on a scale of 0 to 9) in the presence of adherent scales on at least two quarters of the head, and their erythema of the scalp. The 67 subjects were divided into 3 parallel groups of 22 subjects, each group being treated with only one of the two compositions + a control group

Clinical evaluation of the dandruff condition of the scalp (scores) The effects on the films were tested and evaluated by comparing the groups on D2, D7, D15, D21 and D28 through clinical evaluations carried out on the following 25 parameters: free dandruff, adherent dander and erythema, and the overall score was calculated. These evaluations were carried out by qualified dermatologists according to the techniques usually used in the field. At each visit, the dandruff condition, free films and adherent scales, was scored for each item by the investigator using a scale of 0-9, a score of 0 being assigned in the absence of one. dandruff condition and a score of 9 being attributed for a very severe dandruff condition.

The results are expressed as the difference between (i) the value of the score before the start of treatment and (ii) the value of the score at the time of each measurement.

Self evaluation of dandruff condition of the scalp by subjects. The effects on dandruff were tested and evaluated by comparing the groups on D2, D7, D15, D21 and D28 through clinical self-evaluations carried out on the subjects themselves. These self-assessments represent "Topic Reviews". The results are expressed by the value of the skin score attributed by the subject himself. Clinical evaluation of scalp itching The effects on scalp itching were tested and assessed by comparison of groups on days 2, 7, 15, 21 and 28 by conventional clinical evaluations. These evaluations were carried out by qualified dermatologists according to the techniques usually employed in the field. At each visit, itching of the scalp was scored by the investigator using a scale of 0 - 9, a score of 0 is assigned in the absence of itching and a score of 9 being awarded in the presence of very severe itching. . The results are expressed as the difference between (i) the score value before the start of treatment and (ii) the score value at the time of each measurement.

Results The results obtained are illustrated in FIGS. 1 to 4. FIG. 1 illustrates the results of the comparison of the effects on the films of (i) a shampoo comprising an isolated bacterial lysate, free from complete fermentation medium, used at 2, 5% by weight (corresponding to 0.1075% solids), (ii) a shampoo comprising an isolated bacterial lysate, free from complete fermentation medium, used at 5% by weight (corresponding to 0.215% solids), and (iii) the standard anti-dandruff shampoo formula. From the comparison of the results of Figure 1, it appears that the two shampoo formulations comprising an isolated bacterial lysate, used at 0.1075% and 0.215% dry extract, each provide an overall score significantly lower than the overall score obtained with the formulation of reference shampoo. therefore, the results of Figure 1 show that shampoo formulations comprising an isolated bacterial lysate, free from the fermentation medium, exert a very weak, even virtually no anti-dandruff effect, in comparison with the reference shampoo formulation . FIG. 2 illustrates the results of the comparison of the effects on the films of (i) a shampoo comprising the active ingredient of the invention at -20% by weight (corresponding to 0.14% of solids), and (ii) the shampoo formula of reference. The results of FIG. 2 show that the shampoo formulation according to the invention has anti-dandruff activity equivalent to that of the reference formula and this with a solids equivalent of 0: 14%, that is, that is, at an active concentration for which a low effect or no effect is observed with comparative shampoo formulations comprising an isolated bacterial lysate, free of complete fermentation medium (see results in Figure 1) . FIG. 3 illustrates the results of the comparison of the effects on the films of (i) a shampoo comprising the active ingredient of the invention at 0.16% solids content, (ü) a shampoo comprising the active ingredient of the invention at 0.32 % dry extract and (iii) the reference shampoo formula, comprising zinc pyrythione at 1% dry extract. The results of FIG. 3 show that a shampoo formulation comprising 0.16% of active solids according to the invention produces an anti-film effect equivalent to the reference shampoo formulation which comprises 1% by weight of active ingredient. conventional (see also statistical study of EXAMPLE 6). . FIG. 4 illustrates the results of the comparison of the effects on the itch of the scalp of (i) a shampoo comprising the active ingredient of the invention at 0.16% dry extract, (ü) a shampoo comprising the active ingredient of the invention at 0.32% dry extract and (iii) the reference shampoo formula, comprising zinc pyrythione at 1% by weight The results of FIG. 4 show that a shampoo formulation comprising 0.16% and 0.32% of extract dry asset according to the invention produces an effect against the itchy scalp equivalent to the formulation of reference shampoo which comprises 1% by weight of conventional active.

EXAMPLE 3: Test to evaluate the maintenance / restoration of innate defenses: Effect on the release of antimicrobial peptides by a monolayer test of keratinocytes: the effects of the products on the expression of markers Pdef 2,4; S100A7, LL37 and RNase7 were measured at the expression level of messenger RNA (mRNA) by the RT-qPCR technique.

10 - Biological model - cell type: normal human epidermal keratinocytes (NHK) culture conditions: 37 ° C, 5% CO2 - culture medium: keratinocyte-SFM supplemented with Epidermal Growth Factor (EGF) 0.25 ng / ml + pituitary extract ( EP) 25 μg / ml + Gentamycin 25 μg / ml - test medium: keratinocyte-SFM supplemented with Gentamycin 25 μg / ml

Culture and Treatment Normal human epidermal keratinocyte cells (NHEK) were cultivated in culture medium for 30 minutes with the various dilutions tested. After incubation, the cells were rinsed 4 times in PBS and then incubated in a test medium for 72 hours. At the end of the incubation, the culture supernatants were removed, the cell mats were rinsed with PBS solution and immediately frozen at -80 ° C.

Test References The reference tested for BDEF4, Si 00A7 and RNAse 7 is a mixture of cytokines (OSM, TNF-α, IL-17), 30 μg / ml stock solution. The reference tested for CAMP is vitamin D3 (Sigma D1530), 10 -4M.

Quantitative PCR Polymerase chain reaction (PCR) reactions were performed by quantitative PCR with the "Lie Cycler" system (Roche Molecular Systems Inc.). This analysis system makes it possible to carry out fast and efficient PCR reactions, a preliminary development of the conditions of analysis of the different primers. It consists of two main components optimized thermocycler: allowing extremely fast heat transfer. a fluorimeter: to measure continuously the fluorescence intensity incorporated in the DNA (detection at 521 nm). Paired probes specific for the genes studied were used allowing the amplification of the following specific fragments, described in Table 4 below. Table 4 Gene name (protein Abbreviation Gene Bank cDNA encoded pb) Glyceraldehyde-3-phosphate GAPDH NM 002046 269 dehydrogenase - Cathelicidin antimicrobial peptide CAMP NM 004345 251 (LL37) - S100 calcium binding protein A7 SIOOA7 NM 002963 226 Ribonuclease, RNase A family The reaction mixture (10 μg final) for each sample is as follows: 2.5 μg of cDNA - Primers of the different markers used - Reaction mixture (Roche) containing the reaction mixture (10 μg) Taq DNA polymerase enzyme, SR Green I tag (fluorophore that intercalates in double-stranded DNA, during the elongation step) and MgCl 2.

Treatment of Quantitative PCR Data: Incorporation of fluorescence into the amplified DNA is measured continuously during PCR cycles. These measurements make it possible to obtain fluorescence intensity curves as a function of PCR cycles and thus to evaluate a relative expression value for each marker. The number of cycles is determined from the exit points of the fluorescence curves. For the same marker analyzed, the more the sample leaves "late", the lower the initial number of copies of the mRNA. The value of ER (relative expression) is expressed in arbitrary units according to the formula (1/2 number of cycles) x 106. -Standardization of the observed effects on gene expression For a standardized interpretation, the following classification table has been retained 10 (see Table 5 below). Table 5 Classification table% Relative expression versus Class of effect observed in control> 300% Strong stimulation> 200% and <300% Net stimulation> 150% and <200% Moderate stimulation> 50% and <65% Moderate inhibition> 30% and <50% Net inhibition> 150% and <200% Strong inhibition The expressions relating to the markers are presented in Table 1 below. The results are related to the amount of GAPGH messenger (reference gene) and expressed as a% of the untreated control.

Assets tested Complete culture at 7.5 g / l obtained according to Example 1, the complete culture concentrated (150 g / l). A preliminary cytotoxicity test is performed. A preliminary cytotoxicity study by an MTT assay (living cell counting method, well known to those skilled in the art, using the reagent 3- (4,5-dimethylthiazol-2-yl) -2,5-bromide diphenyl tetrazolium) was performed to define the non-cytotoxic test concentrations. Results The effect of the compound on the relative expression of Bdef and RNase7 markers in normal human epidermal keratinocytes. The results are shown in Tables 6 and 7 below.

Table 6 active concentration j3def 4 Control 100 reference: mixture of 10 ng / ml 788321 cytokines Complete culture at 7.5g / 1% (ES = 0.0075%) 234 Table 7 active concentration (3def 4 RNase7% / control% / control Control 100 100 Cytokine mixture 3 ng / ml 607791 407 Complete culture 0.133% 1500 162 concentrated 150g11 (ES = 0.02%) 1726 188 0.4% (ES-0.06%) The products cause a net to strong stimulation of the expression of fidef 4 and RNASE 7.

EXAMPLE 5 MIC negative data to show that the effect is not antifungal Contact the product to be tested and the Malassezia suspension. Place the mixture on the surface of the agar medium. Spread and recover the excess before incubation. Incubate for at least 5 days at 30 ° C.

The products are dissolved in liquid modified Leeming and Notman (LNm) and the tests are performed in duplicate. The complete culture being at 7.5g / l or 150g / l (concentrated version) in dry matter. The complete culture was tested at 10% (for concentration at 7.5g / l) and at 1% (for concentration at 150g / l). The positive reference is pyrithione zinc 1%. The product solutions are twice as concentrated to account for the% 2 dilution when placed in contact with the Malassezia suspension. Table 8 Strain OD Concentrations 550nm cfu / ml Malassezia globosa CBS 7708 1,369 7500 Malassezia restricta CBS 7708 1,137,600 Strains Malassezia globosa and restricta were received on agar slopes and were transplanted and incubated at 30 ° until testing. The suspensions are made in 15 ml of a buffered solution (ref AEB611294, AES). The density of the suspensions is taken at 550 nm. The antifungal effect of the reference product is evaluated by the absence of growth of the strain Malassezia tested. This inhibition is evaluated with respect to the growth control. Inhibitions are noted from 3 to 0 by assessing the culture density on the agar surface in comparison with the growth control of the strain. TABLE 9 RATING INTERPRETATION 3 No growth 2 Growth <in the control box 1 Growth < in the control box 0 Growth comparable to the control box Table 10 Malassezia Malassezia restricta globosa Growth control Extended culture Extended culture suitable density suitable density Pyrithione zinc indicator 3 3 Total inhibition Total inhibition TESTS (Duplicate) B1 B2 B1 B2 TRIS 0 , 1M 50% 0 0 0 0 Complete culture at 10% 0 0 0 0 (concentration 7.5g / 1) Or an ES = 0.075% Complete culture at 1% 0 0 0 0 (concentration 150g / 1) Or an ES = 0.15 The products show no antifungal inhibition on Malassezia restricta and Malassezia globosa. EXAMPLE 6 Effect on the dandruff condition: Opinion on the overall dandruff score and the opinion of the dandruff subjects In this example, the anti-dandruff effect of a shampoo formulation according to the present invention was evaluated. invention, compared with the reference shampoo formulation (comprising zinc pyrythione active). The lysate of Vitreoscilla filiformis bacteria in the complete fermentation medium was used in evapoconcentrated form at lysate solids concentrations of 0.16% and 0.32% ES. Clinical Evaluation of the Dandruff Condition of the Scalp (Scores) The effects on the films were tested and evaluated by comparing the groups at D0, D7, D15, D21 and D28 through clinical evaluations carried out on the following parameters: free dandruff, adherent dander and erythema, and the overall score was calculated. These evaluations were carried out by qualified dermatologists according to the techniques usually employed in the field. At each visit, the dandruff condition, free films and adherent scales, was scored, for each item, by the investigator using a scale of 0 - 9, a score of 0 being assigned in the absence of a state. dandruff and a score of 9 being attributed for a very severe dandruff condition. Self-evaluations were also requested from the subjects of the different groups. These self-evaluations are tréportées under the name "Opinions of the subjects". The results are shown in FIGS. 5 to 9. In FIGS. 5, 6, 8 and 9, the results are expressed in absolute values of the score at each measurement. In Fig. 7, the results are expressed as the difference between (i) the score value before the start of treatment and (ii) the score value at the time of each measurement. FIG. 5 illustrates the comparative overall score results obtained using a shampoo formulation according to the invention comprising 0.16% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), relative to the total weight of the dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of ZnPT active, relative to the total weight of the dry extract. FIG. 6 illustrates the comparative overall score results obtained using a shampoo formulation according to the invention comprising 0.32% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), relative to the total weight of the dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of ZnPT active, relative to the total weight of the dry extract. FIG. 8 illustrates the comparative results of the opinions of the treated patients obtained by using a shampoo formulation according to the invention comprising 0.16% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), relative to the weight total dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of ZnPT active, relative to the total weight of the dry extract. FIG. 9 illustrates the comparative results of the opinions of the treated patients obtained by using a shampoo formulation according to the invention comprising 0.32% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), relative to the weight total dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of ZnPT active, relative to the total weight of the dry extract. FIG. 7 illustrates the comparative results of the overall score differences obtained by using (i) using a shampoo formulation according to the invention comprising 0.16% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), relative to the total weight of the dry extract of the formulation and (ii) using a shampoo formulation according to the invention comprising 0.32% by weight of the active ingredient (bacterial lysate in the complete fermentation medium), by relative to the total weight of the dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of active ingredient, relative to the total weight of the dry extract. All the results of Example 6 show that a shampoo formulation according to the invention, which comprises 0.16% of anti-dandruff active, has an activity at least identical, and even slightly greater, at the same time. reference shampoo formulation which comprises 1% of a conventional anti-dandruff active All the results of example 6 show that a shampoo formulation according to the invention, which comprises 0.32% of anti-dandruff active ingredient. dandruff, has an activity at least identical, and even slightly superior, to the formulation of reference shampoo which includes 1% of a conventional anti-dandruff active Statistical analysis The complete culture at 0.32% ES is never significantly different from ZnPt at 1% ES. Once adjusted to take into account the multiplicity of tests (3 comparisons), the complete culture at 0.16% ES is never significantly different from ZnPt at 1% ES, the two complete culture formulations at 0.32% ES and 0.16% ES. are not significantly different.

EXAMPLE 7 Effect on the dandruff condition: notice of the subjects on the itching In this example, the anti-dandruff effect of a shampoo formulation according to the invention was evaluated in comparison with the reference shampoo formulation. (including zinc pyrythione active). The lysate of Vitreoscilla filiformis bacteria in the complete fermentation medium was used in evapoconcentrated form at lysate solids concentrations of 0.16% and 0.32% ES.

The effects on itchy scalp were tested and assessed by comparison of groups on D2, D7, D15, D21 and D28 through conventional clinical evaluations. These evaluations were carried out by qualified dermatologists according to the techniques usually employed in the field. The results are shown in FIGS. 10 and 11. FIG. 10 illustrates the comparative results of scalp itch effects (view of the subjects) obtained using a shampoo formulation according to the invention comprising 0.32% by weight. weight of the active ingredient (bacterial lysate in the complete fermentation medium), based on the total weight of the dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of active ingredient, relative to the total weight of the dry extract. FIG. 11 illustrates the comparative results of scalp itch effects (view of subjects) obtained using a shampoo formulation according to the invention comprising 0.16% by weight of the active ingredient (bacterial lysate in the body). complete fermentation medium), relative to the total weight of the dry extract of the formulation. The comparative comparative formulation comprises 1% by weight of active ingredient, relative to the total weight of the dry extract.

Claims (10)

REVENDICATIONS1. Cosmetic use of a bacterial lysate (s) in CLAIMS1. Cosmetic use of a lysate of bacterium (s) in a complete fermentation medium, said lysate in its complete fermentation medium being able to induce or stimulate the production of antimicrobial peptides in the scalp, as an active ingredient for preventing and and / or treat the dandruff conditions of the scalp in particular associated with a proliferation of pathogenic microorganisms on the scalp and / or an imbalance of the ecoflora of the scalp. 2. Use according to claim 1 wherein the complete fermentation medium is all or part, in terms of quantity, of the fermentation medium used for the fermentation of said bacterium and in which it was carried out consecutively to its cell lysis. Use according to any one of the preceding claims wherein the lysate mixture and complete fermentation medium contain the cytoplasmic fractions, cell wall fragments and metabolites formed and / or released upon cell lysis of said bacterium and metabolites. soluble and spontaneously released by the bacterium during its fermentation process. 4. Use according to any one of the preceding claims wherein said bacterium is the strain Vitreoscilla filiformis 5. Use according to any one of the preceding claims wherein said bacterium is the strain Vitreoscilla filiformis (ATCC 15551). 6. Cosmetic and / or dermatological composition comprising in a physiologically acceptable medium at least one lysate of bacterium (s) belonging to the genus Vitreoscilla sp. (Especially species Vitreoscilla filiformis), in a complete fermentation medium, said lysate in the complete medium being included in said cosmetic composition in an amount ranging from 0.001% to 0% by weight, relative to the total weight of dry extract of said composition. 7. Composition according to claim 6 being in the form of a hair lotion, a shampoo or a conditioner. 8. Cosmetic process for preventing and / or treating the dandruff conditions of the scalp in particular associated with a proliferation of pathogenic microorganisms on the scalp and / or an imbalance of the ecoflora of the scalp. in an individual, comprising at least one step of administering to said individual at least one lysate of bacterium (s) .2773699 43 belonging to the genus Vitreoscilla sp. (in particular species: Vitreoscilla filiformis or Vitreoscilla beggiatoe) in a complete fermentation medium. 9. Method according to the preceding claim wherein said bacterium is as defined in claims 4 and 5: 5 10. Method according to one of claims 8 and 9 implemented topically.