FR2998174A1 - Preparing topical cosmetic/dermatological active ingredient used in composition for treating acne, comprises culturing microorganism on culture medium as it promotes the formation of supernatant resulting from metabolism of microorganism - Google Patents

Abstract

Preparing a topical cosmetic or dermatological active ingredient containing a supernatant of culture of microorganism comprises: culturing the microorganism on a culture medium as it promotes the formation of the supernatant of a mixture of compounds resulting from metabolism of the microorganism, where the mixture has an microbiocide or microbiostatic activity on human skin microorganisms responsible for nuisances such as body odors, films and/or of film and/or disorders of skin, nails, hair and/or mucous membranes; and recovering the mixture to obtain the active ingredient. Preparing a topical cosmetic or dermatological active ingredient containing a supernatant of culture of microorganism including bacteria, mushrooms, yeasts and microalgae, comprises: culturing the microorganism on a culture medium as it promotes the formation of the supernatant of a mixture of compounds resulting from metabolism of the microorganism, where the mixture has an microbiocide or microbiostatic activity on human skin microorganisms responsible for nuisances such as body odors, films and/or of film and/or disorders of skin, nails, hair and/or mucous membranes; and recovering the mixture to obtain the active ingredient. An independent claim is included for a cosmetic or dermatological composition including the active ingredients. ACTIVITY : Antiseborrheic; Dermatological. MECHANISM OF ACTION : None given.

Description

The invention relates to the use of a mixture of compounds derived from the metabolism of microorganisms, having a microbiocidal or microbiostatic activity, in cosmetics or dermatology. More specifically, it relates to a process for obtaining an active ingredient for the treatment of various conditions of the skin, hair, nails, mucous membranes and hairs, or inconveniences such as body odor and dandruff, the origin of which is linked to the presence of a microbial agent, said active ingredient being derived from a culture of at least one microorganism, in particular bacterial.

Cosmetic or dermatological compositions comprising an active principle of bacterial origin are known. Thus, WO2010 / 013179A1 describes a composition for treating oily skin and related disorders such as acne, this composition comprising a lactic bacterium of the genus Lactobacillus or Bifidobacterium, or a metabolite of this bacterium having the same function. The authors describe its stimulatory activity of skin defense mechanisms, promoting the synthesis of proteins, such as enzymes, involved in these mechanisms. According to W02011 / 073437A1, there is described a deodorant composition, anti-dandruff, care or cleansing of the skin, including oily skin, including acne, whose active ingredient is the combination of a bacteriocin and one or more oligosaccharides or prebiotic polysaccharides. The above compositions contain an active ingredient of bacterial origin, which is either the microorganism itself or its fractions playing the same role, or one or more peptide compounds synthesized naturally by bacteria and used in the isolated state. . The authors of the present invention have discovered that the mixture of the extracellular products of the metabolism of a microorganism, when it is cultivated on a suitable medium, has a direct microbiocidal or microbiostatic activity with respect to certain microorganisms of the microflora. Transient or permanent human skin. This product which constitutes an active principle of the invention has many advantages. First, it does not include the living source microorganism, so it is a stable product; then, it does not require any complex step of treatment and isolation of an active compound; and finally, it acts directly on the target microbial population, so its effect is immediate.

The subject of the invention is thus a process for the preparation of a cosmetic or dermatological active ingredient for topical use, based on a culture supernatant of at least one microorganism, comprising the following steps: a microorganism is available; The microorganism is cultivated on a culture medium such that it promotes the extracellular formation in the culture supernatant of a mixture of compounds derived from the metabolism of the microorganism, said mixture having a directly microbiocidal or microbiostatic activity on at least one microorganism of the cutaneous human microflora causing discomfort such as body odor, dandruff and / or disorders of the skin, nails, hair, hair and / or mucous membranes; and recovering said mixture to obtain said active ingredient. Before discussing the invention in more detail, certain concepts and certain terms employed are hereinafter specified or defined. In the present description, the invention is more oriented towards the use of a bacterium, but of course its scope is not restricted, and, by microorganisms, includes fungi, yeasts, microalgae, which The microbiocidal or microbiostatic activity can be evaluated in the same way as that of a bacterium, since the desired effect is identical. The directly microbiocidal or microbiostatic activity of a compound or a mixture of compounds, as understood according to the invention, means an activity involving an action on the development of the target microorganisms, either via a direct effect compounds on the microbial cells inducing a deterioration of their integrity and / or the inhibition of certain molecular mechanisms, either via a deterioration of the microbial environment leading to the death of the microorganism or by preventing its multiplication, for example by modifying the pH by depleting the medium to an essential component ... This action is mainly dependent on the concentration of the mixture of compounds. By mixing compounds with microbiocidal or microbiostatic activity constituting the active ingredient of the invention, it is understood that this activity is that of the mixture, and not necessarily that resulting from the addition of each of the microbiocidal or microbiostatic activities of the compounds. Thus, this mixture may comprise one or more compounds which, in the isolated state, have little or no microbiocidal or microbiostatic activity, but which within the mixture may exhibit such activity and / or generate or stimulate the microbiocidal activity. or microbiostatic other compounds of the mixture.

This mixture is the product of the metabolism of the microorganism used, that is to say of the microbiological transformation of the constituents of the culture medium selected into secreted compounds in the culture medium and participating in the desired effect. It therefore comprises compounds of low molecular weight and in particular compounds of protein origin, organic acids, nucleic acids, amino acids, sugars, peptides, lipids, and, more generally, compounds derived from primary and secondary metabolism of the microorganism. In an advantageous embodiment of the invention, the microbiocidal or microbiostatic microorganism is chosen from microorganisms classified as QPS (Qualified Presumption of Safety), the QPS classification being a recognition of the harmlessness of QPS microorganisms for humans, by the European Food Safety Authority (European Food Safety Authority Journal 2011; 9 (12): 2497 (1-81). Among the QPS microorganisms, the following microbial genera are advantageously retained: Bifidobacterium, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Pro pionibacterium, Streptococcus, Debaryomyces, Kluyveromyces, Saccharomyces, Hanseniaspora, Schizosaccharomyces, Xantophyllomyces, and more preferably, it is selected among microorganisms classified QPS belonging to the group of so-called lactic bacteria which include the following genera: Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Streptococcus as well as to the genera Propionibacterium and Bifidoba Among these, the bacterial genera Lactobacillus, Lactococcus, Propionibacterium and Bifidobacterium are still preferred.

The active principle of the invention relates to the treatment of skin conditions and inconveniences found directly or indirectly a microbial origin; it can also be effective for the treatment of disorders affecting the mucous membranes and integuments, in particular nails, hair and hair. Thus, the target microorganism (s) are those belonging to the transient or permanent cutaneous flora. The invention is of particular interest when it targets the following species: species of the genus Corynebacterium, in particular Corynebacterium xerosis and Corynebacterium jeikium, species of the genus Brevibacterium, in particular Brevibacterium epidermidis, Brevibacterium casei, Brevibacterium linens, the species of the genus Staphylococcus, in particular Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, the species of the genus Micrococcus, in particular Micrococcus luteus and Micrococcus lylae, the species of the genus Propionibacterium, in particular Propionibacterium acnes, Propionibacterium granulosum, Propionibacterium avidum, species of the genus Malassezia, and in particular Malassezia furfur, - as well as species of the genera Dermabacter, Streptococcus, Acinetobacter, Escherichia, Pseudomonas and Candida. The microbiocidal or microbiostatic effect of the mixture obtained according to the invention results from the metabolism of the microorganism in the culture medium in which it is developed. Culture media suitable for the growth of a microorganism, for example a bacterium, generally comprise yeast extracts, peptones, salts, inorganic or organic sources of phosphate, nitrogen and potassium, etc. . as well as sugars; these commercially available media, as well as the growth conditions of a microorganism (pH, temperature, aeration, agitation, redox potential, duration), and in particular of a bacterium, are well-known notions of the human of the job. According to the invention, such a medium can be developed by those skilled in the art, it can be used as commercially available or can be modified, most often by modifying the concentration and / or the nature of the aforementioned ingredients, to obtain a medium that will promote the production by the microorganism of a mixture of microbiocidal or microbiostatic compounds, secreted in the culture medium. Those skilled in the art are able to determine if the effect is obtained, and therefore able to select a culture medium to produce the desired supernatant. In one of the following examples, a protocol is described which a person skilled in the art can use to evaluate the bactericidal action of a mixture present in a culture supernatant of a bacterium and resulting from the metabolism of the latter, in the culture medium.

Of course, one skilled in the art can evaluate this action by any other method. For optimum production of the microbiocidal or microbiostatic mixture, the microorganism, for example the bacterium, is advantageously cultivated up to an advanced stage of the exponential phase of the microbial growth, preferably in the stationary phase, where the efficiency of the mixture is the stronger, then we recover the mixture. The recovery step of the mixture is conventionally carried out and the routine techniques in microbiology are appropriate. Thus, the supernatant can be separated from the culture medium by filtration or centrifugation, after which the supernatant is treated in order to obtain the active principle, by any suitable technique, notably using concentration, ultrafiltration, drying, lyophilization, atomization, etc. The invention also relates to a cosmetic or dermatological active ingredient for topical use, obtainable by the method of the invention as described above, and a cosmetic or dermatological composition for topical use, comprising one or more such active ingredient. Advantageously, it comprises an active ingredient obtained by cultivation of the Lactococcus lactis strain deposited on November 15, 2012 in accordance with the Budapest Treaty of the National Collection of Microorganisms Culture (CNCM), under the number CNCM I-4693, and is deodorant. or prevents acne: as is demonstrated in the following examples, the cultivation of this strain on a medium MRS or M20, modified or not, produces a supernatant having a bactericidal effect vis-à-vis Brevibacterium epidermis and casei, im plicated in the unpleasant odors that emerge from the feet as well as with respect to Propionibacterium species involved in the development of acne. Another subject of the invention is a cosmetic or dermatological use for topical use of one or more of these active principles. As described in detail in the examples, the authors identified strains, and in particular a strain of the species Lactococcus lactis, and selected a culture medium for producing an active mixture having a bactericidal effect against of bacteria of the cutaneous human microflora, and more precisely with respect to Brevibacterium bacteria responsible for the body odors and Propionibacterium involved in the development of acne. This strain of Lactococcus lactis was deposited at the National Collection of Microorganism Cultures (CNCM) of the Pasteur Institute, depositary authority under the Budapest Treaty and under the accession number of the deposit CNCM 1-4693. A medium that is particularly suitable for obtaining an active active ingredient that is effective against Brevibacterium and in particular Brevibacterium epidermidis is chosen from the commercially available M20 and MRS media, as well as those mediums whose sugar, namely glucose, has been replaced by another. sugar, or whose concentration has been increased. A preferred glucose replacement sugar is sucrose. Another object of the invention is therefore a deodorant composition or acne control or prevention composition comprising an active ingredient obtained by culturing the strain Lactococcus lactis CNCM 1-4693. The compositions according to the present invention may be formulated in any dosage form suitable for their administration. The compositions according to the present invention can thus be formulated as a cream, gel, lotion, milk, oil-in-water emulsion or water-in-oil, solution, ointment, spray, body oil, aftershave, soap, protective baton. lipstick, stick and pencil for make-up, aerosol, roll-on, stick, ball, powder, wipe, incorporation into liposome-type vectors, glycospheres, cyclodextrins, in chylomicrons, macro-, micro-, nano-particles as well as as macro-, micro- and nanocapsules and also adsorption on organic powdery polymers, talc, bentonites and other mineral supports. The compositions according to the present invention may also contain additives or adjuvants customary in cosmetology, such as, for example, other antimicrobial agents or perfumes, but also extraction or synthetic lipids, gelling and viscosifying polymers, tensio- active agents and emulsifiers, hydro- or liposoluble active ingredients, plant extracts, tissue extracts, marine extracts, synthetic actives. The compositions according to the present invention may also comprise other complementary active ingredients chosen for their action, for example for the slimming effect, the anti-cellulite effect, the firming effect, the moisturizing effect, the anti-cellulite effect or the anti-cellulite effect. age, antimicrobial activity, antioxidant activity, anti-radical activity, healing effect, tensor effect, anti-wrinkle effect, chelating activity, complexing and sequestering activity, the effect soothing, anti-dark circles effect, anti-redness effect, emollient activity, hair conditioning effect, anti-dandruff activity, stimulating effect of hair regrowth, anti-hair loss effect of hair, capillary gaining effect, depilatory activity, hair regrowth limiting activity, the activity involved in cell renewal, the activity modulating the inflammatory response, the activity involved in maintaining the oval of the hair. face, but also sunscreen, the anti-irritant activity, cellular nutrition, cellular respiration, antiseborrheic treatments, cutaneous tonicity, protection of the hair. When the compositions according to the present invention contain additional active ingredients, these are generally present in the composition at a sufficiently high concentration so that they can exert their activity. The compositions according to the present invention are preferably used daily and applied one or more times per day. The compositions according to the present invention are very well tolerated, they have no toxicity and their application to the skin, for prolonged periods of time, does not imply any systemic effect. The present invention is now illustrated, but not limited to, by the following examples in support of Figs. 1-6, in which: Fig. 1 is representative photographs of the overlay test of the tested bacterial strains of which Brevibacterium casei is the target. A denotes the control box without streaks of bacteria tested and seeded with an inoculum of Brevibacterium casei; B denotes the box with streaks of a strain of Bifidobacterium adolescentis and seeded with an inoculum of Brevibacterium casei (category -); C denotes the box with streaks of a strain of Propionibacterium freudenreichii and seeded with an inoculum of Brevibacterium casei (category +); D denotes the box with streaks of a strain of Propionibacterium acidopropionici and seeded with an inoculum of Brevibacterium casei (category ++); E denotes the box with streaks of a strain of Lactobacillus plantarum and seeded with an inoculum of Brevibacterium casei (category +++). Figure 2 corresponds to representative photographs of the direct confrontation test on TS medium of which Brevibacterium casei is the target.

A denotes the box seeded with an inoculum of Brevibacterium casei with streaks of Lactococcus lactis CNCM 1-4693; B denotes the box seeded with an inoculum of Brevibacterium casei with streaks of the Lactobacillus plantarum strain above. Figure 3 is representative photographs of the Brevibacterium casei inhibition test with culture supernatants. A denotes the box seeded with an inoculum of Brevibacterium casei with a supernatant of Lactococcus lactis CNCM 1-4693; B denotes the box seeded with an inoculum of Brevibacterium casei with a supernatant of the Lactobacillus plantarum strain above.

Figure 4 shows the comparison of the growth curve of Lactococcus lactis CNCM I-4693 in MRS medium with the inhibitory power of its supernatant on Brevibacterium casei and Brevibacterium epidermidis, where - corresponds to the evolution of the inhibitory power of the Lactococcus supernatant lactis CNCM 1-4693 on Brevibacterium epidermidis, corresponds to the evolution of the inhibitory power of the supernatant of Lactococcus lactis CNCM I-4693 on Brevibacterium casei, and * corresponds to the growth curve of Lactococcus lactis CNCM I-4693 on MRS medium. 5 shows the evolution of the inhibitory power of Lactococcus lactis supernatants CNCM I-4693 on Brevibacterium epidermidis, for different concentrations of the medium in glucose, with - corresponds to the evolution of the inhibitory power of the Lactococcus lactis supernatant CNCM I-4693 grown in M20 medium at 5 g / L of initial glucose, corresponds to the evolution of the inhibitory power of the Lactococcus lactis supernatant CNCM I-4693 grown in M20 medium at 20 g / L of initial glucose, and * corresponds to evolution of the inhibitory power of Lactococcus lactis supernatant CNCM 1-4693 grown in M20 medium at 40 g / l of initial glucose.

The inhibitory activity is expressed in% relative to a reference value. This is the maximum inhibition obtained with the supernatants of the culture of Lactococcus lactis CNCM I-4693 in M20 medium at 5 g / L of glucose (100%).

FIG. 6 shows the evolution of the inhibitory power of Lactococcus lactis CNCM I-4693 supernatants on Brevibacterium epidermidis, for different sugars or sugar concentrations, with - corresponds to the evolution of the inhibitory power of the Lactococcus lactis supernatant CNCM I-4693 grown in M20 medium at 5 g / L of initial glucose, corresponds to the evolution of the inhibitory power of the Lactococcus lactis supernatant CNCM I-4693 grown in M20 medium at 20 g / l of initial glucose, and 30 * corresponds to the evolution of the inhibitory power of the Lactococcus lactis supernatant CNCM I-4693 grown in M20 medium at 20 g / l of initial sucrose. The inhibitory activity is expressed in% relative to a reference value. This is the maximum inhibition obtained with the supernatants of the culture of Lactococcus lactis CNCM I-4693 in M20 medium at 5 g / L of glucose (100%). In the following examples, the following commercially available culture media were used: MRS: Peptone 10g / L Meat Extract 10g / L Yeast Extract 5g / L 10 Glucose 20g / L Tween 80 1.08 g / L Dipotassium phosphate 2g / L Sodium acetate 5g / L Ammonium citrate 2g / L Magnesium sulphate 0.2g / L Manganese sulphate 0.05g / L M20: Tryptone USP 30g / L 20 Yeast extract 20g Glucose 5 g / L It is from these culture media used in the state or modified that bactericidal / bacteriostatic supernatants were obtained, in the examples which follow. EXAMPLE 1 Protocols for Evaluating the Bactericidal / Bacteriostatic Action of a Mixture of the Invention The minimum inhibitory activity is a reference measure for evaluating the efficacy of an antimicrobial agent. In sterile tubes, 3 ml of culture medium containing 105 CFU / ml of the target bacterium is contacted with 1 ml of PBS buffer solution, containing different concentrations of potentially bactericidal or bacteriostatic microorganism supernatant. Controls are carried out in the presence of PBS without addition of culture supernatant. These 35 negative controls make it possible to demonstrate that the conditions used for the test allow the growth of the target bacterium. The tubes are then incubated at a temperature suitable for growth of the target bacterium until the control tubes become turbid and then the absorbance at 600 nm is measured to detect or not the development of the target bacterium. If the culture medium of tubes containing potentially bactericidal or bacteriostatic microorganism supernatant is not cloudy and OD600 is unchanged between T = 0 and the end of the test, it is concluded that growth inhibition has occurred. of the target bacterium and therefore bactericidal or bacteriostatic action of the supernatants tested. The above method is hereinafter applied to the bactericidal evaluation of supernatants derived from culturing Lactococcus lactis CNCM I4693 on M20 medium with 20 g / L of sucrose with respect to Brevibacterium epidermidis. The culture medium used for Brevibacterium epidermidis was TS medium, the tubes were placed 72h at 37 ° C. The minimum activity of inhibition of the supernatant is 0.4-0.6% vis-à-vis Brevibacterium epidermidis.

This supernatant is also active against other harmful microorganisms of the skin, identified in Table 1 below. Table 1: Minimum inhibition activity of Lactococcus lactis culture supernatant CNCM 1-4693 with respect to different microbial species causing skin discomfort Target bacterium Disease / Disease Bactericidal action Brevibacterium epidermidis Unpleasant odor of feet 0 , 4 - 0.6% Propionibacterium acnes Acne vulgaris 0.2 - 0.4% Staphylococcus aureus Infection 6% Example 2: Screening strains of bacteria tested for their antibacterial activity 25 2.1. Inhibition by direct contact between Brevibacterium casei and the screened strains The overlay method used for this screening is one of the many methods for evaluating the inhibitory activities described in the literature (Toure, Kheadr, Lacroix, MoroniFliss: Production of antibacterial 30 substances by bifidobacterial isolates from active stool against Listeria monocytogenes, Journal of Applied Microbiology, 95, pp.1058-1069, 2003).

Fourteen bacterial strains listed in the following Table 2 and classified as QPS were screened according to this method with respect to their antibacterial activity against Brevibacterium casei, a target microorganism. Table 2: Summary of microorganisms screened.

Genus Species Lactobacillus Plantarum Acidophilus Johnsonii Rhamnosus Lactococcus Lactis Propionibacterium Acidipropionici Freudenreichii Bifidobacterium Breve Longum Bifidum Adolescentis Two parallel streaks of strains tested are performed on a suitable agar medium. After growth of the bacteria under anaerobic conditions, this agar is covered with a second medium inoculated with Brevibacterium casei.

The Petri dishes are then aerobically incubated to allow the growth of the target microorganism and then examined to evaluate the presence or absence of growth inhibition zones of Brevibacterium casei around the streaks. After 72 hours of incubation of control boxes without streaking strains tested, we observe a homogeneous development of Brevibacterium casei throughout the agar medium (Figure 1A). Some boxes containing streaks also show a homogeneous development of the target strain indicating a lack of inhibitory activity (Figure 1B). The boxes containing streaks of Lactobacillus plantarum show no development of B. casei. This strain seems to totally inhibit the growth of the target microorganism (Figure 1 E). On the other boxes, Brevibacterium casei inhibition zones centered on the two parallel streaks were observed. These zones are more or less extensive, indicating a power of inhibition of the strains tested different. A note of inhibitory power as a function of the size of the non-growth zone has therefore been attributed (Figure 1). Photographs of the boxes representative of the different categories of inhibition are presented in FIGS. 1B to 1E. Table 3 summarizes all the results obtained.

Table 3: Inhibitions of Brevibacterium casei with the overlay test. Strain Name Intensity of Inhibition Lactobacillus planta rum ++ Lactobacillus acidophilus + Lactobacillus johnsonii ++ Lactobacillus planta rum +++ Lactobacillus rhamnosus +++ Lactococcus lactis + Lactococcus lactis 8270 ++ Lactococcus lactis ++ Bifidobacterium breve ++ Bifidobacterium longum ++ Bifidobacterium bifidum Bifidobacterium adolescentis Propionibacterium freudenreichii + Propionibacterium acidipropionici ++ In order to refine the selection, confrontation tests on the same medium between target microorganisms and inhibitor are carried out. For these trials, both strains are grown simultaneously on the same medium, so there is direct competition between the two organisms for their growth. This test is carried out on TS medium inoculated with a culture of Brevibacterium casei and on which two streaks of a culture of QPS strain were made. This test was not possible with Bifidobacterium strains because they are strict anaerobic, a condition that does not allow the growth of Brevibacterium casei. This test proved to be much more selective than the overlay test since only the Lactococcus lactis CNCM I-4693 strain showed an inhibition zone around the two streaks (FIG. 2A). In the other boxes, the development of Brevibacterium casei is comparable to the control box without streaking and no growth of the strains tested is observed (FIG. 2B). 2.2 Inhibition by the culture supernatants In order to evaluate the efficacy of the culture supernatants of the inhibitory strains selected during the first screening, the diffusion method from the wells was used. For this test, the culture supernatants in stationary phase of strains tested are placed in wells made in an agar inoculated with B. casei. After incubation under conditions adapted to the growth of B. casei, the inhibition diameters around the wells are measured. The tests are carried out four times from the same supernatant. This test proved to be very selective since a zone of no growth of Brevibacterium casei was observed only around the wells containing the supernatants of Lactococcus lactis CNCM 1-4693. This inhibition halo is of a diameter of 1.1 cm (Figure 3). These results confirm those obtained with the overlay and confrontation tests on TS medium. The other supernatants tested did not induce any inhibition halo.

The fact that an inhibition halo around the wells containing the Lactococcus lactis CNCM 1-4693 supernatant is observed, suggests that this strain synthesizes and secretes one or more specific antimicrobial compounds.

Example 3 Optimization of the culture conditions of Lactococcus lactis CNCM 1-4693 3.1 Choice of standard culture medium The culture medium and the growth phase can have a significant influence on the amount of antibacterial compounds synthesized by a strain. Also, the antimicrobial activity of Lactococcus lactis CNCM 1-4693 during its growth on MRS medium was measured and the impact of media MRS, M20, M17 and TBSY on this same activity was evaluated. Several species of Brevibacterium are the source of bad odors; also, it was decided to verify, in parallel with these optimization tests, that the Lactococcus lactis CNCM 1-4693 supernatant inhibits another dominant species of Brevibacterium in the skin. The inhibitory power of culture supernatants was tested against Brevibacterium casei and Brevibacterium epidermidis. As shown in Figure 4, assays with culture supernatants show that the size of the Brevibacterium casei and Brevibacterium epidermidis inhibition zones increases during the exponential growth phase of Lactococcus lactis CNCM 1-4693 and remains constant during the stationary phase. This indicates that the inhibitory molecule (s) are synthesized during the exponential phase. To compare the effect of different media, culture supernatants must be recovered in the stationary phase since the inhibitory activity is maximal and stable at this stage. For the inhibition of Brevibacterium casei, at T = 11 h, the culture supernatants in M17 and TBSY medium induced inhibition halos of 0.77 and 1.08 cm which are smaller than those observed with 10 supernatants. culture in M20 medium and MRS (1.17 and 1.13 cm respectively). The difference in size of the inhibition zones as a function of the culture medium was confirmed in a second series of tests carried out with Brevibacterium epidermidis as target microorganism. Two media seem more suitable for the expression of inhibitory activity: M20 and MRS media. M20 medium still shows greater inhibitory activity. Table 4: Inhibition of Brevibacteria (inhibition halo diameter in cm) due to Lactococcus lactis CNCM I-4693 supernatants obtained after growth of the strain in various media after 11 hours of culture. Medium MRS M20 M17 TBSY Brevibacterium casei 1.13 1.17 0.77 1.08 Brevibacterium epidermidis 1.45 1.47 1.12 1.42 3.2. Influence of the nature and concentration of sugars in the culture medium In order to further increase the expression of the antibacterial activity of this strain, the impacts of the concentration and the nature of the sugars on this activity were evaluated. . Influence of the concentration of sugars In this part, three glucose concentrations in the culture medium were tested. The first test is the reference test: it is the culture of Lactococcus lactis CNCM I-4693 in Erlenmeyer flask in M20 medium at 5 g / l of glucose. The other two experiments are carried out in a fermenter with M20 medium at 20 or 40 g / l of glucose. Samples were taken regularly to measure absorbance at 600 nm, dry matter, sugar concentration and to evaluate antibacterial activity against Brevibacterium epidermidis. The supernatants were used to perform the inhibition test on Petri dishes (FIG. 5). To compare the samples with each other (and evaluate our gain in activity during the optimization steps), it was chosen to express the inhibitory activity in% with respect to a reference value. This is the maximum inhibition obtained with the supernatants of the culture of Lactococcus lactis CNCM 1-4693 in M20 medium at 5 g / L of glucose (100%).

The culture supernatants at 20 g / L glucose taken at T = 6 h 30 and T = 8 h 30 (stationary phase) have 125.8 and 127.3% activities. Similar values (128.8 and 125.8%) were obtained with the supernatants taken at the same time in the fermenter initially containing 40 g / L of glucose. The addition of 20 g / L of glucose and the fermentation of the fermentor culture significantly increase the inhibitory activity present in the supernatants while a further increase in the sugar content (40 g / L) has more effect on the production of inhibitory activity or on biomass. Influence of the nature of sugars These tests were carried out in Erlenmeyer flasks in M20 medium, the nature of the sugar of which was modified. The sugars tested are mannose, sucrose, lactose, lactulose and glucose at a concentration of 5 g / L. The results obtained are given in Table 5.

TABLE 5 Influence of the nature of sugar in culture media of Lactococcus lactis CNCM I-4693 on the% inhibition of Brevibacterium epidermidis. Sugar Glucose Mannose Sucrose Lactose Lactulose% inhibition 100.0 102.2 102.2 90.3 95.7 The supernatants that provide the most important Brevibacterium epidermidis inhibition are those obtained after culturing Lactococcus lactis CNCM I-4693 in M20 medium at 5 g / L of mannose (102.5%) and at 5 g / L of sucrose (102.2%). The supernatants obtained after culturing Lactococcus lactis CNCM I-4693 in M20 medium at 5 g / L lactulose or lactose, provide lower inhibitions to the other sugars (95.7 and 90.3% respectively). Mannose, glucose and sucrose turn out to be sugars of choice to obtain the desired activity.

Claims (16)

REVENDICATIONS1. Process for the preparation of a cosmetic or dermatological active ingredient for topical use, based on culture supernatant of at least one microorganism, characterized in that it comprises the following steps: a microorganism chosen from bacteria is available, mushrooms, yeasts and microalgae; The said microorganism is cultivated on a culture medium such that it promotes the extracellular formation in the culture supernatant of a mixture of compounds derived from the metabolism of the microorganism, said mixture having a directly microbiocidal or microbiostatic activity with respect to at least one microorganism of the cutaneous human microflora responsible for inconveniences such as body odor, dandruff and / or disorders of the skin, nails, hair, hair and / or mucous membranes; and recovering said mixture to obtain said active ingredient. 2. Method according to claim 1, characterized in that the microbiocidal or microbiostatic microorganism is selected from microbial species classified QPS (Qualified Presumption of Safety). 3. Process according to claim 2, characterized in that the microbiocidal or microbiostatic microorganism is chosen from the species of the genera Bifidobacterium, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Pro pionibacterium, Streptococcus, Debaryomyces, Kluyveromyces, Saccharomyces, Hanseniaspora and Schizosaccharomyces. Xantophyllomyces. 4. Process according to claim 3, characterized in that the microbiocidal or microbiostatic microorganism is chosen from the species of the genera Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus, Propionibacterium and Bifidobacterium. 5. Process according to claim 4, characterized in that the microbiocidal or microbiostatic microorganism is chosen from the species of the genera Lactobacillus, Lactococcus, Propionibacterium and Bifidobacterium. 6. Process according to any one of claims 1 to 5, characterized in that the microorganism is cultured to the stationary phase and then the mixture is recovered. 7. Process according to any one of Claims 1 to 6, characterized in that the microorganisms of the cutaneous human microflora are chosen from: the species of the genus Corynebacterium, in particular Corynebacterium xerosis and Corynebacterium jeikium, the species of the genus Brevibacterium , in particular Brevibacterium epidermidis, Brevibacterium casei, Brevibacterium linens, species of the genus Staphylococcus, in particular Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, species of the genus Micrococcus, in particular Micrococcus luteus and Micrococcus lylae, species of the genus Propionibacterium. , in particular Propionibacterium acnes, Propionibacterium granulosum, Propionibacterium avidum, species of the genus Malassezia, and in particular Malassezia furfur, as well as species of the genus Dermabacter, Streptococcus, Acinetobacter, Escherichia, Pseudomonas and Candida. 8. Process according to any one of Claims 1 to 6, characterized in that the microorganisms of the cutaneous human microflora are chosen from: the species of the genus Corynebacterium, in particular Corynebacterium xerosis and Corynebacterium jeikium, the species of Brevibacterium, Brevibacterium linens, - species of the genus Staphylococcus, in particular Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, - species of the genus Propionibacterium, in particular Propionibacterium acnes, Propionibacterium granulosum, Propionibacterium avidum, - species of the genus Malassezia, and in particular Malassezia furfur, 9. Process according to any one of Claims 1 to 6, characterized in that the microorganisms of the cutaneous human microflora are chosen from: species of the genus Brevibacterium, in particular Brevibacterium epidermidis, Brevibacterium casei, Brevibacterium linons, species of the genus Propionibacterium, especially Propionibacterium acnes, Propionibacterium granulosum, Propionibacterium avidum, 10. Method according to any one of claims 1 to 9, characterized in that the bacterium is a strain Lactococcus lactis CNCM I10 4693. 11. Method according to any one of claims 1 to 10, characterized in that the culture medium comprises soya peptones (30 g / L), yeast extract (20 g / L) and sucrose ( 20 g / L) 12. Cosmetic or dermatological active ingredient for topical use, obtainable by the method according to any one of claims 1 to 11. 13. Cosmetic or dermatological composition for topical use, comprising one or more active ingredients according to claim 12. 14. A composition according to claim 13, characterized in that it comprises an active principle obtained by culturing the strain Lactococcus lactis CNCM 1-4693, and in that it is deodorant by inhibiting the growth of species of the genus Brevibacterium. 15. A composition according to claim 13, characterized in that it comprises an active ingredient obtained by culturing the strain Lactococcus lactis CNCM 1-4693, and in that it fights or prevents acne by inhibiting the growth of the species. of the genus Propionibacterium. 16. Cosmetic use of one or more active ingredients according to claim 12.