Cosmetic composition for inducing melanin synthesis in individual or animal, method of preparation, cosmetic method and use

Abstract

The present invention describes a composition for inducing melanin synthesis in an individual or animal, including the use for preventing and / or reversing the hair grey process.

Description

Technical Field

[0001] The present invention describes a new composition for inducing melanogenesis in an individual or an animal, including its use for preventing and / or reversing the process of hair graying. The present invention relates to the fields of cosmetics, dermatology and / or pharmaceutical treatment. Background Art

[0002] Hair color, like skin color, is the result of the quantity and type of melanin produced by melanocytes and transferred to keratinocytes. Pigmentation of the hair follicle involves the melanogenic activity of follicular melanocytes, the transfer of melanin granules to cortical and medullary keratinocytes, and the formation of the pigmented hair shaft (SLOMINSKI et al., 2005).

[0003] Hair graying is one of the most obvious indicators of human aging. The regulation of the graying process stems from multiple factors, such as genetics, oxidation, psychological emotions, metabolism, endocrine and nutrition, etc. (O'SULLIVAN et al., 2021).

[0004] More specifically, the appearance of white hair is a phenomenon associated with a decrease in the number of follicular melanocytes and a reduction in melanin content (SARIN, ARTANDI, 2007). Regarding the density of follicular melanocytes, several mechanisms interfere with this factor: the absence or failure of differentiation of melanocyte stem cells, defects in melanocyte migration, melanocyte apoptosis, and malfunction of the pigmentation mechanism (TOBIN, 2011).

[0005] It must be emphasized that, like melanocyte density, the regulation of melanogenesis plays an equally important role. Melanin in the skin and hair follicles is produced within organelles called melanosomes found in the cytoplasm of melanocytes. Melanocytes are derived from immature melanoblasts. During embryonic development, these cells migrate from the neural crest (a transient component of the ectoderm located between the neural tube and the epidermis) to the basal layer of the epidermis and then into the developing hair follicles (SLOMINSKI et al., 2005; TOBIN, 2008). The factors that cause these subsets of neural crest cells to become melanocytes are still poorly understood, but several factors are known to be involved, including MITF (microphthalmia-associated transcription factor), SOX10 transcription factor, Pax3 protein, KIT, FGF-2 (fibroblast growth factor-2), endothelin 3, etc. (TOBIN, 2008).

[0006] In the hair follicle, the melanin produced is transferred from the melanocytes to the keratinocytes of the hair bulb (SLOMINSKI et al., 2005). Melanin is produced in a complex pathway that involves a series of chemical and enzymatic reactions known as melanogenesis. The color of an individual's hair is the result of tyrosinase (TYR) activity, and thus its regulation and activity are crucial during melanin synthesis (TOBIN, 2008). In addition to tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2), also known as dopachrome tautomerase (DCT), are also important mediators of this process (PILLAIYAR et al., 2017).

[0007] Melanogenesis begins with the oxidation of tyrosine to dopa and the oxidation of dopa to dopaquinone through the action of TYR in the presence of molecular oxygen. This first step is the rate-limiting step of melanin synthesis because the remaining reactions can occur spontaneously at physiological pH (PILLAIYAR et al., 2017). From this moment on, the presence or absence of cysteine will determine which pigment (eumelanin or pheomelanin) is formed.

[0008] In the absence of cysteine, dopaquinone is converted to leucodopachrome and then to dopachrome by auto-oxidation. Then, dopachrome can produce 5,6-dihydroxyindole (DHI) by spontaneous decarboxylation, and DHI rapidly oxidizes and polymerizes to form an insoluble, dark brown-black high molecular weight polymer, which belongs to eumelanin. However, if TRP-2 / DCT is available, dopachrome can produce 5,6-dihydroxy-2-carboxylic acid (DHICA) through the action of this enzyme, and DHICA will also form eumelanin through the action of TRP-1. In the presence of cysteine, dopaquinone reacts with this molecule to form cysteine-dopa, which then oxidizes and polymerizes to produce a soluble red-yellow pigment, collectively known as pheomelanin (PILLAIYAR et al., 2017).

[0009] Melanogenesis can be regulated through multiple pathways. It is important to emphasize that the stimulation of this process is mainly caused by proopiomelanocortin peptide (POMC) in keratinocytes. After the production and subsequent cleavage of this molecule, different products are generated, including melanocyte-stimulating hormone (α-MSH), adrenocorticotropic hormone (ACTH), β-lipotropin (β-LPH), β-endorphin (β-END), as well as β-MSH and γ-MSH. ACTH and α-MSH are considered to be some of the main stimulators of melanogenesis (PILLAIYAR et al., 2017; SCHERER et al., 2010).

[0010] Different strategies have been developed to reverse the process of graying.

[0011] It has been reported that iron supplementation in anemic patients reverses follicular hypopigmentation associated with certain nutrient deficiencies (BHAT et al., 2013). Topical treatments are available on the market and may contain various compounds (p-aminobenzoic acid, phytic acid, Capixyl, calcium pantothenate, etc.), but the results are often inconsistent and only temporary (SEHRAWAT et al., 2017). The basis of the treatment is usually to disguise gray hair with dyes, and some studies have shown that hair dyes can protect hair from light damage (SEHRAWAT et al., 2017). More promising treatments include anti-aging or antioxidant compounds, recombinant human growth hormone treatment, and the use of α-melanocyte-stimulating hormone (α-MSH) agonists such as Greyverse TM (glycerol + water + palmitoyl tetrapeptide-20 amide-IFF Lucas Meyer) - patent document AU2008271875A1. As a result of activating the MC1R receptor, Greyverse TM increases the synthesis of the receptor itself, indicating the core mechanism of action of this active ingredient.

[0012] Another product example is the combination of dihydroquercetin glucoside and acetyl tyrosine (Darkenyl TM - Givaudan) - patent document WO2020089216A1. According to the results presented, this product induces the gene expression of genes such as MC1R and POMC in a statistically significant manner. Another commercially available topical cosmetic for combating graying is EUK-134 (ethylbisiminomethyl guaiacol manganese chloride - Lucas Meyer Cosmetics). The latter consists of mimetics of two natural skin enzymes (superoxide dismutase and catalase), which play an important antioxidant role and thus more precisely combat oxidative stress, but do not act on stimulating melanin synthesis.

[0013] Other patent documents can be cited as describing inventions aimed at reversing graying.

[0014] CN102988235 describes a combination of antler, red ginseng (Radix Ginseng Rubra), fleeceflower root (Radix Polygoni Multiflori), black soybean (Semen sojae atricolor), and Brassica Campestris oil that can promote natural hair repigmentation.

[0015] KR102265875 describes a composition aimed at using exosomes derived from stem cells to darken hair and prevent graying.

[0016] WO2014096086 describes the topical use of the lysate form of the microorganism Bifidobacterium longum for pro-pigmentation effects.

[0017] PI0916690 (WO2010013182) discloses the oral cosmetic use of the probiotic microorganism (i.e., live microorganism) or its lysate of Lactobacillus paracasei for the cosmetic treatment of oily scalp diseases (especially dandruff).

[0018] KR102570587 further describes the use of a product obtained by fermenting a microorganism of the genus Lactobacillus for preventing hair graying or improving gray hair, said product being a mixture obtained by fermenting a culture medium containing an energy source (such as skim milk powder) with a Lactobacillus. None of the above documents describe the specific species of the microorganism tested and the species of the microorganism that produced the experimental results, only contenting themselves with saying that it is a culture selected from a total of 4,000 species of Lactobacillus (Example 1). In addition, it generally describes that the strain can be one of Lactobacillus pentosus, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus acidophilus. According to the patent disclosure, the fermentation product must contain a large amount of carboxyl groups, which react with the amino acids in the hair strand at room temperature, changing the color of the hair strand through a reaction outside the fiber. Therefore, its product is not only different in nature from the present invention, but also a reagent that chemically acts on the hair strand externally.

[0019] In addition, according to a search of the prior art, several examples of products with antimelanogenic effects produced using compositions and / or extracts of Lactobacillus were found. For example, as described by Lim, HY et al. in "Antiwrinkle and Antimelanogenesis Effects of Tyndallized Lactobacillus acidophilus KCCM12625P", Int. J. Mol. Sci., 2020, 21, 1620, which used Lactobacillus acidophilus; Choi, WS et al., "Whitening Effects of Lactobacillus rhamnosus Associated with Its Antioxidative Activities", Korean J. Microbiol. Biotechnol. (2013), 41(2), 183–189, which used Lactobacillus rhamnosus and / or Lactobacillus casei; Bajpai, VK et al., "Partially Purified Exo-Polysaccharide from Lactobacillus Sakei Probio65 with Antioxidant, α-Glucosidase and Tyrosinase Inhibitory Potential", J. Food Biochemistry, 40(3), (2016), 264-274, which used Lactobacillus sakei; Kim HR et al., "Lipoteichoic acid isolated from Lactobacillus plantarum inhibits melanogenesis in B16F10 mouse melanoma cells", Mol Cells, 2015, 38(2):163-70, which used Lactobacillus plantarum.

[0020] This indicates a high degree of inherent unpredictability in this technical field, where seemingly similar products may not necessarily be equally effective.

[0021] Therefore, this patent application aims to propose a new method to solve the problems of the prior art. It can be inferred from the studied literature that no document predicts or implies the teachings of the present invention. Summary of the Invention

[0023] Therefore, the present invention aims to solve the problems in the prior art through a new composition that surprisingly has a melanogenic effect.

[0024] In addition to reducing the expression of MC1R, the composition of the present invention also shows the result of inhibiting the expression of the POM gene. Both the MC1R and POM genes are important biomarkers in the process of melanogenesis and thus also important biomarkers for hair pigmentation. Furthermore, the composition of the present invention can actively regulate the expression of other genes (such as TYR, DCT, TYRP-1, MITF, MYO5A, PMEL, MLPH), indicating that its mechanism of action is different from that of the prior art.

[0025] Therefore, a new cosmetic composition is provided for inducing melanogenesis in an individual or an animal, and the cosmetic composition contains at least 0.0001 wt% of the lysate of the microorganism Lacticaseibacillus paracasei.

[0026] In one embodiment of the present invention, the composition contains 0.001 - 20 wt% of the lysate.

[0027] In one embodiment of the present invention, the composition contains 0.001 - 5 wt% of the lysate.

[0028] In one embodiment of the present invention, the composition is used for preventing and / or reversing the graying process of hair filaments.

[0029] In one embodiment of the present invention, the composition is used for inducing melanogenesis in hair follicles.

[0030] In one embodiment of the present invention, the composition is a topical hair cosmetic composition.

[0031] A new method for producing the above composition is also proposed, and the method includes:

[0032] - Anaerobically culturing the microorganism Lacticaseibacillus paracasei in a culture medium until the stationary phase is reached;

[0033] - Inactivating and lysing the microorganism completely or partially by placing the culture medium at a temperature of 55 - 80 °C for at least 30 minutes;

[0034] - Filtering and centrifuging;

[0035] - Mix the resulting lysate with at least one suitable cosmetic vehicle.

[0036] There is also provided a cosmetic method for inducing melanogenesis in an individual or animal to prevent and / or reverse the process of hair graying, the method comprising topically applying to hair follicles a composition as described above in a cosmetically suitable amount that allows for said induction in the individual or animal.

[0037] There is also provided the use of a composition as described above in the preparation of a cosmetic composition for inducing melanogenesis in an individual or animal to prevent and / or reverse the process of hair graying.

[0038] In one embodiment of the invention, the composition is for application to a skin area comprising hair follicles.

[0039] These and other objects of the invention will be directly recognized by those skilled in the art, and a sufficiently detailed description will be provided below to enable those skilled in the art to reproduce it.

[0040] Brief Description of the Drawings

[0041] In order to better define and clarify the content of this patent application, the following figures are given:

[0042] Figure 1 : Representative images of the effect of Lacticaseibacillus paracasei lysate on melanogenesis in ex vivo hair follicles - Warthin Starry staining (objective x40), compared with the same preparation and a reference composition without the lysate (placebo). Detailed Description of the Invention

[0044] The detailed description of the invention establishes some non-limiting definitions of the main terms and technical features used in this patent application and provides examples of some embodiments of the invention so that those skilled in the art can reproduce it.

[0045] Thus, unexpectedly, the inventors observed that certain microorganisms of the species Lacticaseibacillus paracasei are particularly effective in preventing the process of hair graying and reversing its effects.

[0046] Thus, based on the data provided in the examples, the inventors characterized the ability of Lacticaseibacillus paracasei lysate to stimulate the expression of a large number of genes related to melanogenesis in hair follicles, and in addition, directly demonstrated the stimulation of melanin production in hair follicle cultures.

[0047] The lysate of Lacticaseibacillus paracasei can stimulate the gene expression of tyrosinase (TYR), dopachrome tautomerase (DCT), tyrosinase-related protein-1 (TYRP-1), melanogenesis-associated transcription factor (MITF), myosin 5A (MYO5A), premelanosome protein (PMEL), and melanophilin (MLPH), and all these genes are related to the pigmentation process in hair follicles and skin. In addition, in the hair follicle culture experiment, the inventors of the present invention demonstrated that the lysate of Lacticaseibacillus paracasei can significantly increase melanogenesis in the hair bulb.

[0048] Therefore, the application of the cosmetic, dermatological or pharmaceutical composition of the present invention promotes the prevention and / or reversal of the graying process of hair.

[0049] According to another aspect of the present invention, the present invention relates to the use of Lacticaseibacillus paracasei lysate or its fraction (part) in the preparation of a pharmaceutical and / or dermatological composition, and the composition is intended for the prevention and / or reversal of the graying process of hair.

[0050] In the context of the present invention, the term "prevention" refers to reducing the risk of the occurrence of the phenomenon under discussion.

[0051] According to another aspect of the present invention, the present invention relates to a cosmetic method for treating and / or preventing the graying process, which includes at least one step of administering Lacticaseibacillus paracasei lysate or its fraction to an individual in an amount effective for the treatment and / or prevention under discussion.

[0052] According to another aspect of the present invention, the lysate is topically administered to an area containing hair follicles.

[0053] Microorganism

[0054] As previously mentioned, the Lacticaseibacillus paracasei microorganism used as the active composition proposed in this patent application (also known as Lactobacillus paracasei, the previous taxonomic definition of this microorganism, and updated according to the systematic classification of the genus Lactobacillus described by Zheng et al. in the professional journal "International Journal of Systematic and Evolutionary Microbiology" in 2020) is used in the form of a lysate.

[0055] A lysate generally refers to a substance obtained by disrupting or lysing biological cells through a phenomenon called cell lysis, resulting in the release of intracellular biological components naturally contained in the cells of the microorganism under discussion.

[0056] In the context of the present invention, the term "lysate" is used to denote the whole lysate or only a fraction (part) thereof obtained by lysis of the microorganism in question. Thus, the present invention relates to the use of a lysate of Lactobacillus paracasei and / or a fraction thereof for the purposes described herein.

[0057] Thus, the lysate used is formed entirely or in part by intracellular biological components and / or components of the cell wall and cell membrane. Such cell lysis can be carried out by various techniques, such as osmotic shock, heat shock, sonication or under mechanical stress such as centrifugation.

[0058] According to a preferred embodiment of the present invention, the lysate is obtained by carrying out cell lysis at 55 - 80 °C and centrifuging at the end of the process.

[0059] In one example, the microorganism Lactobacillus paracasei is grown anaerobically in a suitable medium. When the stationary phase of development is reached, the medium is inactivated and lysed, for example by heating to a temperature of 55 - 80 °C (optionally to 60 - 65 °C) for 30 minutes by a pasteurization process. The microorganisms are then removed by conventional separation techniques (such as membrane filtration, centrifugation), and the supernatant is subsequently used as the product.

[0060] The active ingredients that form the lysate and belong to Lactobacillus paracasei can be formulated into a composition at a concentration of at least 0.0001% (expressed as dry weight), in particular 0.001 - 20%, more particularly 0.01 - 5%, based on the dry weight of the active substance, relative to the total weight of the vehicle or relative to the total weight of the composition containing it.

[0061] The compositions of the present invention can be present in all galenic forms that are generally suitable for the adopted administration method.

[0062] Depending on the type of composition considered, the carrier / vehicle can have different properties.

[0063] More specifically, for compositions for topical administration, these can be aqueous, water - alcoholic or oily solutions, solution - type dispersions or emulsion - type or serum - type dispersions, emulsions of liquid or semi - liquid consistency obtained by dispersing the oil phase in the water phase (O / W) or emulsions of liquid or semi - liquid consistency obtained by dispersing the water phase in the oil phase (W / O), or suspensions or emulsions of soft, semi - solid or solid consistency, creams of aqueous or anhydrous gel type, or microemulsions, microcapsules, microparticles or vesicle dispersions of ionic and / or non - ionic type.

[0064] In one embodiment of the present invention, the product formulation can be benzyl alcohol: 0.80%, ethylhexylglycerol: 0.20%, and lysate of Lactobacillus paracasei: q.s. to 100%.

[0065] These compositions can particularly form creams for cleaning, protecting, treating or caring for the scalp, lotions, gels or mousses for caring for the scalp, such as cleansing or disinfecting lotions or bath compositions. They can also be in the form of solutions, creams, gels, lotions, mousses or in the form of aerosol compositions also containing a pressurized propellant for use on the scalp. Specifically, they can be in the form of hair conditioners, hair rinses, leave-in products, shampoos, etc.

[0066] The above ingredients with anti-gray hair effects have the characteristics that make them suitable for different types of hair care products with different compositions, such as:

[0067] Alcohol-free hair conditioner, a composition containing: 0.10% xylitol sesquicaprylate, 2% PPG-5-ceteth 20, 10% Lactobacillus paracasei lysate, and water q.s. to 100%.

[0068] Alcohol-containing hair conditioner, a composition containing: 0.10% xylitol sesquicaprylate, 20% ethanol 96%, 2% PPG-5-ceteth 20, 10% Lactobacillus paracasei lysate, and water q.s. to 100%.

[0069] Hair rinse, a composition containing: 0.10% disodium EDTA, 1% glycerin, 0.50% cetyltrimethylammonium chloride, 0.22% lactic acid 85%, 3.00% cetearyl alcohol 30 / 70, 1.00% cetyl alcohol, 0.20% stearamidopropyl dimethylamine, 0.50% xylitol sesquicaprylate, 2% Lactobacillus paracasei lysate, and water q.s. to 100%.

[0070] Leave-in product, a composition containing: 0.10% disodium EDTA, 1% glycerin, 0.80% cetyltrimethylammonium chloride, 0.10% citric acid 50%, 0.22% lactic acid 85%, 3.00% cetearyl alcohol 30 / 70, 0.70% cetyl alcohol, 0.30% ethoxylated cetearyl alcohol 20EO, 0.50% xylitol sesquicaprylate, 2% Lactobacillus paracasei lysate, and water q.s. to 100%.

[0071] Shampoo, a composition containing: 4% cocamidopropyl betaine, 27% sodium lauryl ether sulfate, 3% sodium lauroyl sarcosinate, 1% lauryl glucoside, 4% cocamide DEA, 0.50% xylitol sesquicaprylate, appropriate amount of sodium chloride, 50% citric acid solution q.s. to pH = 6.00 - 7.00, 2% Lactobacillus paracasei lysate, and water q.s. to 100%.

[0072] It is well known that Galenic forms for topical application can also contain additives commonly used in the cosmetic, pharmaceutical and / or dermatological fields, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active ingredients, preservatives, antioxidants, solvents, fragrances, fillers, filters, odor absorbers and colorants. The amounts of these various additives are those commonly used in the fields in question, for example 0.01 - 20% of the total weight of the composition. Depending on their nature, these additives can be introduced into the fatty / oil phase and / or the aqueous phase.

[0073] The products can be used as cosmetic, dermatological or pharmaceutical compositions for topical use, which are capable of regulating the production of melanin in hair follicles. These compositions can in particular constitute creams, lotions, gels, shampoos, hair conditioners or mousses for the care of the scalp, such as cleansing or disinfecting lotions or bath compositions.

[0074] They can also be used on the scalp in the form of solutions, creams, gels, lotions, mousses or in the form of aerosol compositions which also contain a pressurized propellant.

[0075] Example 1: Evaluation of the regulation of gene expression in human skin fragments

[0076] Materials and methods

[0077] Skin explants (n = 6) obtained from surgical operations carried out in an ophthalmic plastic surgery clinic were cut into 0.4 cm 2 . The fragments obtained were transferred to cell culture plates containing a medium and treated with a preparation containing Lactobacillus paracasei lysate (3% w / w) and the same preparation without said lysate (placebo). After treatment, the fragments were stored for 24 hours in a humid atmosphere at 37 °C, 5% CO 2 and then collected in RNAlaterTM solution (Qiagen).

[0078] After culturing with the preparation for 24 hours and storing in RNAlaterTM solution (Qiagen), the ex vivo tissues were mechanically macerated and then RNA was extracted, quantified and analyzed for its integrity using a commercially available kit.

[0079] Using a High-Capacity cDNA Reverse A kit (Applied Biosystems) was used for cDNA synthesis on a StepOnePlus device (Applied Biosystems). Subsequently, the cDNA was quantified on a Nanodrop Lite device (Thermo Scientific).

[0080] High-throughput RT-qPCR was performed using the Biomark HD system (Fluidigm, San Francisco, CA) associated with the IFC controller HX that supports the IFC 96.96 Dynamic Array TM Finally, the cycle threshold was determined specifically for each assay using Fluidigm software, and the expression results were analyzed using the delta-delta Ct method. TM

[0081] Results

[0082] The expression of the same genes in skin fragments treated with Lactobacillus paracasei lysate (3% w / w) was compared with those treated only with the formulation without the lysate (placebo).

[0083] Comparison between Lactobacillus paracasei lysate and placebo group

[0084]

[0085] Table 1: Percentage regulation of relative gene expression in skin fragments treated with a formulation containing Lactobacillus paracasei lysate (3%) compared to the placebo group

[0086] Example 2: Evaluation of the induction of melanogenesis in hair follicle culture

[0087] Materials and methods

[0088] Hair follicles were dissected and inoculated in a 24-well plate (1 hair follicle / well) in a medium containing Lactobacillus paracasei lysate (0.002% w / w) or without Lactobacillus paracasei lysate (control) and a comparative composition (benchmark = glycerol, water, and palmitoyl tetrapeptide-20-amide 0.002% w / w). For each test condition, an excess of hair follicles was prepared to reach the target value of 12 high-quality hair follicles for each condition at the end of the experiment. The hair follicles were cultured for 7 days, and the treatment was renewed by removing and replacing the medium at 3 and 5 days of culture.

[0089] At the end of the culture, the hair follicles were fixed with a formaldehyde solution. Then the hair follicles were dehydrated and subsequently embedded in paraffin. Cross-sections (5 μm thick) were made using a microtome.

[0090] ​The cross-sections were dewaxed and then subjected to Warthin Starry staining. Finally, the cross-sections were observed under a microscope (40x objective lens) and images were recorded. The amount of melanin was evaluated by measuring the labeled area of melanin in the hair follicle bulb normalized by the hair follicle bulb area (semi-quantitative image analysis method).

[0091] Results

[0092] Compared with the control group (untreated), the Lactobacillus paracasei lysate increased the melanin level by 43%. Under the same conditions, the reference product increased the melanin level in the hair follicles by 14% ( Figure 1 ).

[0093] The Lactobacillus paracasei lysate was able to strongly stimulate melanin synthesis in the hair follicle bulb. Quantification of the pigmented area showed a 43% increase compared to the control group, while under the same experimental conditions, the reference was able to stimulate 14% pigmentation.

[0094] The embodiments disclosed herein are intended to illustrate only some of the various implementations and usage forms of the present invention, without limiting the interpretation of its scope and breadth, as well as possible alternative forms and configuration changes defined based on the claims presented herein.

Claims

1. A cosmetic composition for inducing melanin synthesis in an individual or an animal, characterized in that, the composition comprises at least 0.0001 wt% of a lysate of the microorganism Lactobacillus paracasei.

2. The composition according to claim 1, characterized in that, the composition comprises 0.001 - 20 wt% of the lysate.

3. The composition according to claim 1, characterized in that, the composition comprises 0.001 - 5 wt% of the lysate.

4. The composition according to claim 1, 2 or 3, characterized in that, the composition is for preventing and / or reversing the graying process of hair filaments.

5. The composition according to any one of the preceding claims, characterized in that, the induction of melanin synthesis is carried out in hair follicles.

6. The composition according to any one of the preceding claims, characterized in that, the composition is a topical cosmetic composition.

7. A method for preparing the composition according to any one of the preceding claims, characterized in that, the method comprises: - anaerobically culturing the microorganism Lactobacillus paracasei in a culture medium until the stationary phase is reached; - completely or partially inactivating and lysing the microorganism by placing the culture medium at a temperature of 55 - 80 °C for at least 30 minutes; - filtering and centrifuging; - mixing the resulting lysate with at least one suitable cosmetic vehicle.

8. A beauty method for inducing melanin synthesis in an individual or an animal, characterized in that, the method comprises topically applying, in a cosmetically suitable amount, the composition according to any one of claims 1 - 6 to allow said induction in the individual or the animal.

9. The method according to claim 8, characterized in that, the method is for preventing and / or reversing the graying process of hair filaments, and the topical application is carried out on hair follicles.

10. Use of the composition according to any one of claims 1 - 6, characterized in that, for preparing a cosmetic composition for inducing melanin synthesis in an individual or an animal.

11. The use according to claim 10, characterized in that, the composition is for preventing and / or reversing the graying process of hair filaments, and for application to a skin area containing hair follicles.

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