Cosmetic, nutraceutical and / or dermatological use of a strain of Lactobacillus crispatus and / or a composition comprising it

Abstract

The present invention relates to the cosmetic, nutraceutical use of a strain of Lactobacillus crispatus, in particular that filed under number CNCM I-5579, and / or of a composition comprising it, to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of the skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on the skin and / or mucous membranes and / or detoxify the skin and / or mucous membranes.It also concerns the strain of Lactobacillus crispatus, or a dermatological composition comprising it, for its use, advantageously by topical route, for the treatment and / or prevention of pathologies of the skin and / or mucous membranes associated with an increase in pigmentation of the skin and / or mucous membranes, in particular to depigment and / or prevent hyperpigmentation of pathological skin and / or pathological mucous membranes.

Description

Title of the invention: Cosmetic, nutraceutical and / or dermatological use of a strain of Lactobacillus crispatus and / or a composition comprising it

[0001] The present invention relates to the cosmetic, nutraceutical and / or pharmaceutical use, particularly dermatological, of a strain of Lactobacillus crispatus and / or a composition comprising it to decrease pigmentation of the skin and / or mucous membranes and / or prevent the increase of pigmentation of the skin and / or mucous membranes.

[0002] The structure and appearance of the skin are modified with age and / or under the influence of extrinsic and / or intrinsic factors. These modifications are numerous and include changes in the pigmentation of the skin and / or mucous membranes. The skin becomes dull, less luminous, less radiant, its luminosity diminishes, and the complexion becomes less even, with the appearance of age spots.

[0003] With chronological aging or accelerated aging, particularly under the influence of radiation, typically ultraviolet radiation or pollution, the appearance of these changes, such as age spots, is accelerated. They are linked to the presence of melanin and lipofuscin, the excess and abnormal distribution of which in the skin cause the appearance of brown spots, also known as age spots or pigment spots. These can appear as a marker of accelerated melanogenesis and are often considered unsightly. They give the complexion an uneven and dull appearance.

[0004] Melanogenesis and oxidative stress are the subject of numerous studies and innovations in the field of cosmetics for the development and improvement of cosmetic active ingredients aimed at reducing and / or preventing the increase of pigmentation of healthy skin and / or healthy mucous membranes.

[0005] There is a need in the field of cosmetics and dermatology to develop alternative natural active ingredients capable of reducing pigmentation of the skin and / or mucous membranes and / or preventing the increase of pigmentation of the skin and / or mucous membranes, in particular to preserve and / or improve the complexion of the skin and / or mucous membranes, in particular its radiance and / or luminosity and / or homogeneity, particularly in a population more prone to pigment spots, in particular skin of phototype IV to VI as defined by the Fitzpatrick classification method.

[0006] There is a permanent need to identify agents having one or more of these properties, in particular having proven efficacy and safety.

[0007] In a particularly surprising way, the Applicant discovered that Lacto-bacillus crispatus, a beneficial commensal bacterium of the skin and / or mucous membranes, has the property of reducing pigmentation of the skin and / or mucous membranes and / or preventing its increase, in particular to preserve and / or improve the complexion of the skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on the skin and / or mucous membranes.

[0008] The Applicant also unexpectedly discovered that this bacterium also has the property of detoxifying the skin and / or mucous membranes.

[0009] The Applicant's invention is all the more interesting because the bacterium Lacto-bacillus crispatus, and in particular that filed under number CNCM 1-5579, has the advantage of being naturally present on young skin, unlike the bacteria used until now in cosmetic and / or dermatological treatments.

[0010] This invention is even more interesting because the Applicant has demonstrated that with age and / or under the influence of intrinsic and / or extrinsic factors, the bacterium Lactobacillus crispatus is less present and less abundant on the skin. Thus, the object of the present invention has the advantage of rebalancing and / or supplementing the skin and / or mucous membrane microbiota, in particular with a bacterium of the skin flora that diminishes and becomes less abundant over time and / or with exposure to intrinsic and / or extrinsic factors.

[0011] The object of the present invention is therefore particularly suitable for the treatment of so-called mature skin, especially of people over 50 years of age.

[0012] The strain according to the invention also has the advantage of being an ingredient that is easy to formulate and can be easily manufactured on an industrial scale. It is a topically acceptable active ingredient for the skin and mucous membranes, and does not present a risk of allergy.

[0013] Applications have already described the use of Lactobacillus crispatus in cosmetics. Application WO9822082 thus discloses the use of sphingomyelinase that can be extracted from lactic acid bacteria for topical application to increase ceramide levels in the skin and mucous membranes, for the prevention or therapeutic treatment of several pathologies.

[0014] Application WO2009031099 discloses the use of a probiotic to prevent the onset and / or to treat the manifestation of sensations of discomfort and / or skin signs associated with a surface skin treatment or an invasive treatment, for aesthetic purposes.

[0015] Application WO2019111189 describes the use including a probiotic in par Lactobacillus paracasei specifically for the prevention of inflammatory damage induced by UV radiation.

[0016] However, none of these applications discloses or suggests the cosmetic, nutraceutical and / or pharmaceutical, particularly dermatological, use of a strain of Lactobacillus crispatus and / or a composition comprising it to reduce pigmentation of the skin and / or mucous membranes, and / or to prevent its increase, in particular to preserve and / or improve the complexion of the skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on the skin and / or mucous membranes.

[0017] Furthermore, none of these applications discloses or suggests the use of a strain of Lactobacillus crispatus and / or a composition comprising it to detoxify the skin and / or mucous membranes.

[0018] The present invention has as its primary object the cosmetic and / or nutraceutical use of a strain of Lactobacillus crispatus, preferably the Lactobacillus crispatus strain filed under number CNCM 1-5579, to reduce pigmentation of the skin and / or mucous membranes and / or to prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or healthy mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigmentary spots on healthy skin and / or healthy mucous membranes, preferably brown spots and / or age spots, and / or to detoxify healthy skin and / or healthy mucous membranes, in particular skin of phototype IV to VI as defined by the Fitzpatrick classification method.

[0019] According to a particular embodiment of this use, the strain according to the invention decreases and / or inhibits melanogenesis, in particular by acting on the decrease and / or inhibition of melanin, preferentially on the decrease and / or inhibition of the synthesis and / or quantity of melanin, and / or by decreasing and / or inhibiting the activity of tyrosinase, and / or by detoxifying healthy skin and / or healthy mucous membranes.

[0020] According to one aspect of the invention, the strain according to the invention can be used to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes. The strain according to the invention can also be used to preserve and / or improve the radiance and / or luminosity and / or evenness of the complexion of healthy skin and / or healthy mucous membranes. The strain according to the invention can further be used to prevent the appearance and / or reduce the presence of skin pigmentation spots healthy skin and / or mucous membranes, preferably brown spots and / or age spots. The strain according to the invention can also be used to detoxify healthy skin and / or mucous membranes, in particular through antioxidant action and / or action on lipid peroxidation and / or by increasing and / or stimulating glutathione synthesis induced by oxidative stress.

[0021] It also relates to a cosmetic treatment method characterized in that it comprises the topical application to at least one area of ​​healthy skin and / or healthy mucous membrane of a strain of Lactobacillus crispatus according to the invention, preferably the Lactobacillus crispatus strain CNCM 1-5579, or of a cosmetic composition comprising it, to reduce pigmentation of the skin and / or mucous membranes and / or to prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or healthy mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on healthy skin and / or healthy mucous membranes, and / or to detoxify the skin and / or mucous membranes.

[0022] The invention further relates to a strain of Lactobacillus crispatus according to the invention, preferably the Lactobacillus crispatus strain CNCM 1-5579, or to a pharmaceutical composition, in particular a dermatological one comprising it, for the prevention and / or treatment of skin and / or mucous membrane pathologies associated with increased pigmentation of the skin and / or mucous membranes, in particular to depigment and / or prevent hyperpigmentation of pathological skin and / or mucous membranes, to decrease pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of pathological skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of pathological skin and / or mucous membranes,and / or to prevent the appearance and / or reduce the presence of pigment spots on pathological skin and / or pathological mucous membranes and / or to detoxify pathological skin and / or pathological mucous membranes.

[0023] For the purposes of the present invention, "cosmetic use" means a non-pharmaceutical use, that is to say, one which is not intended for therapeutic use and is applied to a part of the body said to be healthy, in particular to an area of ​​the skin and / or mucous membranes said to be healthy.

[0024] For the purposes of the present invention, "nutraceutical use" means a use intended for non-pharmaceutical oral use, i.e., one which does not require therapeutic treatment.

[0025] The term "skin" means any area of ​​skin on any part of the body and / or face, including the scalp.

[0026] For the purposes of the present invention, "phototype IV to VI skin" refers to specific skin types classified according to the Fitzpatrick scale, which classifies individuals based on the reaction of their skin and hair to UV radiation, and / or classified as such by a dermatologist based on skin tone, which depends in particular on the level of melanocyte activity, the creation of these cells, and their grouping. Preferably, these dark skin types of phototype IV to VI originate from characteristic ethnic backgrounds, respectively from India, Asia, and / or Africa. In particular,

[0027] - Phototype IV (Mediterranean type) corresponds to a fair, matte skin and Brown or black hair and eyes. Sunburns are rare, skin tans well, a dark brown;

[0028] - Phototype V (Latino type) corresponds to fair, matte skin, hair and black eyes. Sunburns are rare, skin tans well, a dark brown;

[0029] - Phototype VI (African type) corresponds to very dark skin, hair and Dark eyes. Sunburns are very rare.

[0030] According to the invention, the term "mucous membrane(s)" refers to the ocular mucous membrane, the nasal mucous membrane, the auricular mucous membrane, the vaginal mucous membrane, the urogenital mucous membrane, and / or the oral mucosa, particularly the buccal, labial, and / or gingival mucosa, preferably the ocular and / or oral mucosa, and even more preferably the labial and / or ocular mucosa. Advantageously, it does not include the vaginal mucosa.

[0031] For the purposes of the present invention, "healthy skin" or "healthy mucous membrane" means an area of ​​skin or mucous membrane on which the strain according to the invention is applied and which is said to be "non-pathological" by a dermatologist, that is to say, not showing any infection, scar, disease, inflammation or skin condition such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, or any wounds or injuries.

[0032] For the purposes of the present invention, "Lactobacillus crispatus strain" means the bacterium Lactobacillus crispatus in whole or partial form, in particular as a lysate, fraction, or metabolite. Advantageously, the Lactobacillus crispatus strain is not genetically modified.

[0033] The Lactobacillus crispatus strain according to the invention may be derived from any known species of Lactobacillus crispatus. Preferably, the Lactobacillus crispatus strain is the species deposited under the Budapest Treaty with the Pasteur Institute (28 rue du Docteur Roux, F-75024 Paris cedex 15) on 09 / 09 / 2020 under the designation CNCM 1-5579.

[0034] The advantage of this particular strain of Lactobacillus crispatus is that it is the first species of Lactobacillus crispatus found naturally on the skin.

[0035] For the purposes of the present invention, "reducing skin and / or mucous membrane pigmentation and / or preventing its increase" means depigmenting, whitening, lightening, preventing the increase, maintaining and / or preserving and / or improving the complexion and / or skin tone of the skin and / or mucous membranes, and preferably reducing and / or preventing the increase of melanogenesis, and even more preferably reducing and / or totally or partially inhibiting gene and / or protein expression and / or tyrosinase activity and / or reducing and / or inhibiting the synthesis and / or quantity of melanin.

[0036] The skin pigmentation process is also called "melanogenesis" and comprises numerous steps, from the synthesis of melanin in melanocytes to its transport to the keratinocytes of the epidermis via melanosomes. Several proteins are involved in this process. Tyrosinase is the enzyme that plays a role in key steps of melanin synthesis; it cooperates with other proteins to transform melanin into eumelanin and pheomelanin, the melanins responsible for the pigmentation process.

[0037] “Decrease and / or inhibit melanogenesis” means to reduce and / or inhibit totally or partially gene and / or protein expression and / or tyrosinase activity and / or to reduce and / or inhibit totally or partially the synthesis and / or quantity of melanin.

[0038] According to the invention, "reducing and / or totally or partially inhibiting tyrosinase activity" means causing a decrease of at least 25%, preferably at least 50%, and even more preferably at least 75% of the tyrosinase activity in the presence of the Lactobacillus crispatus strain according to the invention, compared to the level of activity measured in the absence of the extract.

[0039] Preferably, the measurement of tyrosinase activity is carried out on tyrosinase extracted from so-called normal human melanocytes, more preferably by measuring melanin production by photometrically measuring the absorbance at 540 nm from tyrosinase extracted from so-called normal human melanocytes, more preferably in the presence of the Lactobacillus crispatus strain prepared according to Example Le, advantageously under the conditions described in Example 3.

[0040] According to the invention, "totally or partially reducing and / or inhibiting the synthesis and / or quantity of melanin" means a decrease in melanin of at least 10%, preferably at least 20%, and even more preferably at least 30% of the synthesis and / or quantity of melanin in the presence of the Lactobacillus crispatus strain according to the invention, compared to the synthesis and / or quantity of melanin detected in the absence of the strain. In a preferred embodiment of The invention relates to a reduction in the amount of melanin in the presence of the Lactobacillus crispatus strain according to the invention prepared according to example 1a or according to example 2. Preferably, the amount of melanin is measured by optical density at 475 nm from B16 melanocytes cultured in vitro in a culture medium comprising fetal calf serum, advantageously under the conditions described in example 2. In another preferred method, the amount of melanin is measured by optical density at 475 nm from a co-culture of melanocytes and so-called normal human keratinocytes, cultured in vitro in a culture medium comprising growth factors, advantageously under the conditions described in example 4.

[0041] For the purposes of the present invention, "preserve and / or improve the complexion" means to maintain and / or prevent the decrease and / or increase the brightness and / or luminosity and / or radiance and / or brilliance of the complexion and / or the homogeneity of the complexion and / or decrease and / or reduce the dull and / or earthy complexion of the skin and / or mucous membranes.

[0042] The measurement of skin tone radiance, luminosity, and evenness of the complexion of the skin and / or mucous membranes can be studied using measurement techniques known to those skilled in the art, in particular, in vivo, by chromametry, or by image analysis. This latter in vivo measurement method consists of taking high-resolution cross-polarized photographs of the faces of volunteers at a 45° angle before and after application of the product being tested. Based on these digital photographs, image analysis allows the extraction and quantification of specific parameters (e.g., L*, a*, b*, C, h°) related to skin color, radiance, and evenness.

[0043] For the purposes of the present invention, "preserve and / or improve the radiance of the complexion" means to maintain and / or prevent the decrease and / or increase the brightness and / or radiance and / or brilliance and / or clarity, and / or decrease and / or reduce the dull and / or earthy complexion of the skin and / or mucous membranes.

[0044] For the purposes of the present invention, "preserving and / or improving the homogeneity of the complexion" means maintaining and / or preventing the increase and / or decrease in the quantity and / or extent of imperfections and / or irregularities in the color of the complexion, in particular chosen from among redness, pigment spots, comedones, freckles and / or visible pores.

[0045] The homogeneity of skin tone and the amount of melanin in the skin can be measured by mexametry. This in vivo measurement method uses the principle of absorption / reflection to derive a melanin index that reflects the amount of melanin present in the skin.

[0046] Advantageously, the measurement of complexion improvement is carried out on a population of women with pigment spots on the face, preferably tiellement by application of a cream containing the strain of Lactobacillus crispatus prepared according to example 1, under the conditions described in example 7.

[0047] "Maintaining and / or preventing the increase and / or decrease of imperfections and / or irregularities in skin color" means preventing the increase and / or decrease of the amount of melanin in the skin by at least 2%, advantageously by at least 3% in the presence of the strain according to the invention during a 28-day application on the skin compared to the amount of melanin detected in the absence of the strain and / or by at least 2%, advantageously by at least 5% in the presence of the strain according to the invention during a 56-day application on the skin compared to the amount of melanin detected in the absence of the strain.

[0048] "Preventing the appearance and / or reducing the presence of pigment spots" means maintaining and / or preventing the increase and / or reducing the pigmentation of localized pigment spots, in particular the number and / or intensity of pigment spots typically on areas of the face, décolletage, neck, back, shoulders and / or hands, particularly spots on the tops of the hands.

[0049] Pigment spots include, in particular, brown and / or dark and / or white spots and / or "aged spots" when located on a part of the body exposed to UV radiation and / or related to chronological or UV-induced aging, for example, during sun exposure. Pigment spots also include freckles and / or moles and / or post-inflammatory spots and / or spots that appear in response to aggressions, and / or of hormonal origin, particularly in the context of chloasma or melasma, also known as the mask of pregnancy, and / or of drug origin, and / or to prevent and / or treat unsightly pigmentary manifestations of the skin and / or mucous membranes accompanying a non-cutaneous pathology.

[0050] The Applicant also discovered that the strain of Lactobacillus crispatus, in particular that filed under the designation CNCM 1-5579, can be used to detoxify the skin and / or mucous membranes, including healthy skin and / or mucous membranes.

[0051] "Detoxifying the skin and / or mucous membranes" means the prevention and / or total and / or partial inhibition of oxidative stress, in particular by the prevention and / or total and / or partial inhibition of the formation of free radicals, and / or lipid peroxidation and / or by increasing and / or stimulating the synthesis of glutathione, which has antioxidant and detoxifying properties.

[0052] Detoxifying helps to protect the skin against aggressions, both external and internal, which cause oxidative stress likely to impair its proper functioning and appearance, in particular a means capable of stimulating the natural systems of protection and repair of skin cells and limiting the induction of cellular damage.

[0053] “Oxidative stress” refers to the formation of excessive reactive species, y including reactive oxygen species and reactive nitrogen species, which are formed when the skin is subjected to oxidative stress caused by various factors such as solar ultraviolet, infrared and visible light, environmental pollution, including ozone and particulate matter, and psychological stress.

[0054] In addition to impacting the effectiveness of skin detoxification systems, oxidative stress can also exacerbate pigmentation, notably by inducing uneven skin tone or pigmentary disorders.

[0055] In a preferred embodiment of the invention, it is a matter of reducing and / or inhibiting the formation of DPPH° (2,2-diphenyl-l-picrylhydrazyl) free radicals by at least 10%, preferably by at least 30%, in the presence of the Lacto-bacillus crispatus strain according to the invention compared to the formation of DPPH° free radicals detected in the absence of the strain, and / or reducing and / or inhibiting the release of intracellular free radicals induced by oxidative stress, preferably by UVA, by at least 20%, preferably by at least 50%, in the presence of the Lactobacillus crispatus strain according to the invention compared to the release of intracellular free radicals detected in the absence of the strain.

[0056] In a preferred embodiment of the invention, the aim is to reduce and / or inhibit the spontaneous formation of DPPH° free radicals, in the presence of the Lactobacillus crispatus strain according to the invention prepared according to example 1a and / or 1a

[0057] Preferably still, the formation of DPPH° free radicals is measured in tubo by optical density at 530 nm, under the conditions described in example 5.a.

[0058] In a preferred embodiment of the invention, the aim is to reduce and / or inhibit the release of intracellular free radicals induced by oxidative stress, preferably by UVA, in the presence of the Lactobacillus crispatus strain according to the invention prepared according to the example.

[0059] Preferably still, the release of intracellular free radicals induced by oxidative stress, preferably by UVA, is measured by fluorescence emitted by the formation of DCF (2',7-dichlorofluorescein) from the DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) probe at excitation wavelengths of 485 nm and emission wavelengths of 538 nm from normal human fibroblasts, under the conditions described in Example 5.b.

[0060] In another embodiment, it is a matter of "preventing and / or inhibiting the peroxidation of membrane lipids", that is to say, of totally or partially preventing and / or inhibiting lipid oxidation induced by oxidative stress, preferably by UVA of at least 10%, preferably of at least 30% in the presence of the Lactobacillus crispatus strain according to the invention compared to the lipid peroxidation detected in the absence of the strain.

[0061] In a preferred embodiment of the invention, this involves preventing and / or inhibit membrane lipid peroxidation induced by oxidative stress, preferably by UVA, in the presence of the Lactobacillus crispatus strain according to the invention prepared according to example the

[0062] Preferably still, the peroxidation of membrane lipids induced by oxidative stress, preferably by UVA, is measured by fluorescence of the MalonDiAldehde (MDA) formation at excitation wavelengths of 532 nm and emission wavelengths of 560 nm from normal human fibroblasts, under the conditions described in Example 5.c.

[0063] According to another embodiment, it is a matter of maintaining and / or stimulating glutathione synthesis, that is to say, preventing the decrease and / or increasing the expression and / or quantity of glutathione in the presence of oxidative stress, preferably induced by UVA, of at least 20%, preferably of at least 50%, in the presence of the Lactobacillus crispatus strain according to the invention compared to the glutathione synthesis detected in the absence of the strain.

[0064] In a preferred embodiment of the invention, the aim is to maintain and / or stimulate glutathione synthesis induced by oxidative stress, preferably by UVA, in the presence of the Lactobacillus crispatus strain according to the invention prepared according to the example.

[0065] Preferably still, glutathione synthesis is measured in vitro by fluorescence at excitation wavelengths of 355 nm and emission wavelengths of 430 nm from normal human fibroblasts, under the conditions described in Example 6.

[0066] According to the present invention, the Lactobacillus crispatus strain can be used in whole form, in particular viable and / or inactivated, in particular dead, and / or in lysate form, and / or in the form of one or more of its fractions and / or in the form of one or more of its metabolites, preferably from its secretome.

[0067] For each of its forms, the strain according to the invention can be isolated or associated with its culture medium.

[0068] Preferably, the strain according to the invention is used associated with its culture medium, i.e. contained in its culture medium.

[0069] For the purposes of the present invention, "culture medium" means a support containing the nutrients enabling the culture and / or multiplication of the Lactobacillus crispatus strain according to the invention. Preferably, this is an MRS medium (Man, Rogosa and Sharp agar).

[0070] For the purposes of the present invention, "isolated" means not mixed with one or more compound(s) that may be associated with it in its culture medium.

[0071] For the purposes of the present invention, "whole form" means its native form, a form in which the bacterial envelope is intact, as opposed to a lysed form. When used in whole form, the Lactobacillus crispatus strain can be viable and / or inactivated and / or dead.

[0072] For the purposes of the present invention, "viable" means a strain of Lacto-bacillus crispatus according to the present invention capable of forming colonies in culture.

[0073] The production of the viable whole Lactobacillus crispatus strain may be carried out according to any method classically known to those skilled in the art. Advantageously, it will be carried out according to the protocol as described, for example, in Example La.

[0074] For the purposes of the invention, "inactivated" means a strain of Lactobacillus crispatus according to the present invention which is no longer able, temporarily or permanently, to form colonies in culture.

[0075] The inactivation of the Lactobacillus crispatus strain can be carried out using any method conventionally known to those skilled in the art. In particular, it can be inactivated by irradiation, heat treatment or, under certain conditions, by high-pressure treatment or extrusion. Advantageously, it will be carried out by heat treatment, again advantageously according to the protocol as described, for example, in example 1b

[0076] Thermal inactivation can be achieved by incubating the Lactobacillus crispatus strain for a given period of time, advantageously from about 10 seconds to 90 minutes, preferably from about 15 minutes to one hour, and at a temperature advantageously from about 60°C to 150°C. Preferably, it will be incubated for about 30 minutes at about 80°C.

[0077] For the purposes of the invention, "dead" means a strain of Lactobacillus crispatus according to the present invention which is no longer capable, permanently, of forming colonies in culture.

[0078] For the purposes of this invention, "lysate" means a material obtained following the destruction or dissolution of biological cells by a process known as cell lysis, thereby releasing the intracellular biological constituents naturally contained in the cells of the microorganism in question and fragments of cell membrane components. The lysate used is therefore composed of all the intracellular biological constituents and the constituents of the cell walls and membranes of the Lactobacillus crispatus strain according to the invention.

[0079] The lysate can be obtained by any method classically known to those skilled in the art. In particular, it can be obtained by osmotic shock, thermal shock, ultrasound, enzymatic lysis, tyndallization, or under mechanical stress such as centrifugation, or by increasing the pressure or combinations of these different technologies.

[0080] Preferably, the lysate will be obtained by a combination of mechanical action and of increased pressure. Preferably, the lysate will be obtained according to the protocol as described, for example, in example lc

[0081] In a preferred embodiment of the invention, the lysate will be associated with the culture medium of the bacteria.

[0082] For the purposes of the present invention, "fractions" means a fragment of the Lactobacillus crispatus strain according to the present invention, having efficacy in reducing pigmentation of the skin and / or mucous membranes and / or preventing its increase, in particular in preserving and / or improving the complexion of the skin and / or mucous membranes, preferably in preserving and / or improving the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or in preventing the appearance and / or reducing the presence of pigment spots on the skin and / or mucous membranes and / or detoxifying the skin and / or mucous membranes. This fraction corresponds to a non-total portion of the intracellular biological constituents and the constituents of the cell walls and membranes obtained by lysis of the Lactobacillus crispatus strain according to the invention.

[0083] According to a particular embodiment of the invention, the fractions may be the protoplasm of the bacterium. According to a particular embodiment of the invention, the fraction corresponds to all the intracellular biological constituents, but does not contain constituents of the cell walls and membranes of the Lactobacillus crispatus strain.

[0084] In a preferred embodiment of the invention, the fractions will be associated with the culture medium of the bacteria.

[0085] For the purposes of the present invention, "metabolites" means one or more organic and / or inorganic molecules resulting from the metabolism of the Lactobacillus crispatus strain according to the present invention and also possessing efficacy in reducing pigmentation of the skin and / or mucous membranes and / or preventing its increase, in particular in preserving and / or improving the complexion of the skin and / or mucous membranes, preferably in preserving and / or improving the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or in preventing the appearance and / or reducing the presence of pigment spots on the skin and / or mucous membranes and / or detoxifying the skin and / or mucous membranes. Preferably, this refers to the secretome of the Lactobacillus crispatus strain according to the invention.

[0086] In a particular embodiment of the invention, the metabolites will be associated with the culture medium of the bacteria.

[0087] For the purposes of the present invention, "secretome" means the set of organic and / or inorganic molecules secreted by the Lactobacillus crispatus strain according to the present invention, which has the efficacy of reducing pigmentation of the skin and / or mucous membranes and / or preventing its increase, in particular to preserve and / or improve the complexion of the skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of the skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on the skin and / or mucous membranes and / or detoxify the skin and / or mucous membranes.

[0088] The secretome can be obtained by any method classically known to those skilled in the art. Advantageously, it will be obtained according to the protocol as described, for example, in Example Le.

[0089] In a preferred embodiment of the invention, the secretome will be associated with the culture medium of the bacteria.

[0090] According to a preferred embodiment of the invention, the Lactobacillus crispatus strain according to the invention will be used in whole viable form and / or in the form of one or more of its metabolites, preferably its secretome.

[0091] According to an advantageous embodiment, the Lactobacillus crispatus strain according to the invention does not contain sphingomyelinase.

[0092] According to an advantageous embodiment of the invention, the Lactobacillus crispatus strain according to the invention is applied topically to healthy skin and / or healthy mucous membranes. Preferably, the skin and / or mucous membranes are of phototype IV to VI. Phototypes IV to VI are as defined by the Fitzpatrick classification method.

[0093] For the purposes of the present invention, "topical route" means the direct local application and / or vaporization of the Lactobacillus crispatus strain according to the invention or of the composition comprising it according to the invention on the surface of the area of ​​skin and / or mucous membrane concerned.

[0094] According to the invention, the Lactobacillus crispatus strain according to the invention can be used alone, as an active ingredient and / or in a cosmetic and / or nutraceutical, and / or pharmaceutical composition, in particular dermatological, preferably intended for topical and / or oral application, again preferably for topical application.

[0095] When used in whole live form according to the invention, and when used alone, as an active ingredient, the strain according to the invention can be in dry form, i.e. in powder form, advantageously in maltodextrin, preferably in a strain content of 10% to 80% (w / w) by weight of the strain, preferably from 30% to 70% (w / w), further advantageously from 40% to 60% (w / w) by weight of the strain relative to the total weight of powder.

[0096] Alternatively, the whole live form according to the invention can be used dispersed and / or diluted in a solvent. Preferably, this solvent contains less than 20% (v / v) by volume of water, more preferably less than 5% (v / v) by volume of water, and more preferably the solvent contains no water.

[0097] According to an alternative embodiment, this solvent contains less than 20% (w / w) by weight of water, more preferably less than 5% (w / w) by weight of water, and more preferably the solvent contains no water. Most preferably, this solvent is a cosmetically acceptable oil.

[0098] When used in whole inactivated form, in particular dead, and / or in lysate form, and / or in the form of one or more of its fractions and / or in the form of one or more of its metabolites, in particular its secretome, and when used alone as an active ingredient, the strain according to the invention may be in dry form, i.e. in powder form, advantageously in maltodextrin, preferably in a strain content of 1% to 80% (w / w) by weight of the strain, preferably of 3% to 50% (w / w) by weight of the strain, most preferably of about 5% or 20% (w / w) by weight of the strain relative to the total weight of powder, again preferably about 10% (w / w), again preferably 10% (w / w) by weight of the strain relative to the total weight of powder.

[0099] Alternatively, the inactivated whole forms, in particular dead, and / or the lysates, and / or the fractions, and / or the metabolites, in particular the secretome according to the invention, may be used soluble and / or diluted and / or dispersed in a solvent, in particular polar, such as water, an alcohol, a polyol, a glycol such as pentylene glycol and / or hexylene glycol and / or caprylyl glycol and / or butylene glycol, or a mixture thereof, preferably a hydroglycolic or hydroalcoholic mixture, more preferably comprising a glycol selected from hexylene glycol, caprylyl glycol and mixtures thereof.

[0100] In another embodiment, the strain according to the invention can be incorporated into a cosmetic and / or nutraceutical composition comprising at least one cosmetically acceptable and / or nutraceutically acceptable excipient. Preferably, it is incorporated into a cosmetic composition comprising at least one cosmetically acceptable excipient.

[0101] For the purposes of the present invention, a "cosmetically acceptable" excipient means a compound and / or solvent that is topically acceptable, i.e., does not induce an allergic response on contact with the skin, including the human scalp, and / or mucous membranes, and is non-toxic, non-unstable, and chemically.

[0102] In a preferred embodiment of the invention, the strain according to the invention is present in the cosmetic composition, at a concentration of 1x10 4% to 10% (w / w) by weight, preferably between 1x10 4% and 5% (w / w) by weight, more advantageously between 1x10 3% and 0.5% (w / w) by weight, relative to the total weight of the composition, said composition further comprising at least one cosmetically acceptable excipient.

[0103] Particularly advantageously, when the strain according to the invention is used in the form of one or more of its metabolites, in particular its secretome, it is present in the cosmetic composition, at a concentration of 0.05 to 0.5% (w / w) by weight, or preferably at a concentration of about 0.1% (w / w) by weight, relative to the total weight of the composition, said composition also comprising at least one cosmetically acceptable excipient.

[0104] Particularly advantageously, when the strain according to the invention is used in whole viable form, it is present in the cosmetic composition, at a concentration of 0.01 to 0.1% (w / w) by weight, or even more preferably at a concentration of about 0.025% (w / w) by weight, relative to the total weight of the composition, said composition further comprising at least one cosmetically acceptable excipient.

[0105] The cosmetic composition of the invention can be selected from a suspension or solution, aqueous or oily, an aqueous cream or gel or an oily gel, in particular a shower gel, a shampoo; a milk; an emulsion, a microemulsion or a nanoemulsion, in particular oil-in-water or water-in-oil or multiple or silicone; a mask; a serum; a lotion; a liquid soap; a dermatological bar; an ointment; a balm; a butter; a mousse; a patch; an anhydrous product, preferably liquid, pasty or solid, for example in the form of makeup powders, sticks or tablets, in particular in the form of lipstick.

[0106] It may also be a makeup product or a makeup removal product.

[0107] Advantageously, the Lactobacillus crispatus strain or the composition of the invention is intended to be applied to healthy skin and / or healthy mucous membranes of all or part of the body and / or face and / or scalp, preferably the legs, thighs, arms, belly, décolleté, neck, armpits, lips, again preferably all or part of the face, and preferably the cheeks, forehead, chin, lips, eye contour.

[0108] Advantageously, the strain and / or the composition comprising it is intended to be applied to an area of ​​healthy skin with a dull complexion and / or an area of ​​non-homogeneous healthy skin and / or an area of ​​healthy skin with the appearance of pigment spots.

[0109] Preferably, the Lactobacillus crispatus strain according to the invention is particularly suitable for the formulation of a so-called neutral and gentle composition for respecting the sebaceous gland, in particular the skin including the scalp and / or mucous membranes.

[0110] Alternatively, the Lactobacillus crispatus strain according to the invention can be presented in all the galenic forms classically used for oral application, in particular as a nutraceutical active ingredient and / or nutraceutical composition.

[0111] The compositions according to the invention may contain any suitable solvent and / or any suitable vehicle and / or any suitable excipient, possibly in combination with other compounds of interest.

[0112] Therefore, for these compositions, the excipient contains, for example, at least one compound chosen from the group including preservatives, emollients, emulsifiers, surfactants, moisturizers, thickeners, conditioners, mattifying agents, stabilizers, antioxidants, texturizing agents, gloss enhancers, film-forming agents, solubilizers, pigments, colorants, perfumes and sunscreens. These excipients are preferably chosen from the group consisting of amino acids and their derivatives, polyglycerols, esters, polymers and cellulose derivatives, lanolin derivatives, phospholipids, lactoferrins, lactoperoxidases, sucrose-based stabilizers, vitamin E and its derivatives, natural and synthetic waxes, vegetable oils, triglycerides, unsaponifiables, phytosterols, vegetable esters, silicones and their derivatives, protein hydrolysates,Jojoba oil and its derivatives, lipo / water-soluble esters, betaines, aminoxides, plant extracts, sucrose esters, titanium dioxide, glycines, and parabens, and preferably from the group consisting of butylene glycol, steareth-2, steareth-21, glycol-15 stearyl ether, cetearyl alcohol, phenoxyethanol, methylparaben, ethylparaben, propylparaben, butylparaben, butylene glycol, natural tocopherols, glycerin, dihydroxycetyl sodium phosphate, isopropyl hydroxycetyl ether, glycol stearate, triisononanoin, octyl cocoate, polyacrylamide, isoparaffin, laureth-7, a carbomer, propylene glycol, glycerol, bisabolol, a dimethicone, sodium hydroxide, PEG 30-dipolyhydroxysterate, caprylic / capric triglycerides, cetearyl octanoate, dibutyl adipate, grapeseed oil, jojoba oil, magnesium sulfate, EDTA, a cyclomethicone,Xanthan gum, citric acid, sodium lauryl sulfate, mineral waxes and oils, isostearyl isostearate, propylene glycol dipelargonate, propylene glycol isostearate, PEG-8, beeswax, hydrogenated palm kernel oil glycerides, hydrogenated palm oil glycerides, lanolin oil, sesame oil, cetyl lactate, lanolin alcohol, castor oil, titanium dioxide, lactose, sucrose, low-density polyethylene, isotonic saline solution, and mixtures thereof.

[0113] Many cosmetically active ingredients are known to those skilled in the art to improve the health and / or physical appearance of the skin (including the scalp) and / or mucous membranes. Those skilled in the art know how to formulate cosmetic, nutraceutical, or dermatological compositions to obtain the best effects. Moreover, these compounds can have a synergistic effect when combined with each other.to others. These combinations are also covered by the present invention. The CTFA Cosmetic Ingredient Handbook, Second Edition (1992) describes various cosmetic and pharmaceutical ingredients commonly used in the cosmetic and pharmaceutical industries, which are particularly suitable for topical use. Examples of these ingredient classes include, but are not limited to, the following compounds: abrasives, absorbents, and compounds for aesthetic purposes such as perfumes, pigments, colorants, essential oils, astringents, etc.(e.g., clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate), anti-acne agents, anti-flocculants, antifoaming agents, antimicrobial agents (e.g., iodopropyl butylcarbamate), antioxidants, binders, biological additives, buffering agents, swelling agents, chelating agents, additives, biocidal agents, denaturants, thickeners, and vitamins, and derivatives or equivalents thereof, film-forming materials, polymers, opacifying agents, pH adjusters, reducing agents, depigmenting or lightening agents (e.g., hydroxyquinone, kojic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl glucosamine), conditioning agents (for example: humectants).

[0114] The cosmetic composition may also include other cosmetic and / or nutraceutical agents having the same properties and inducing a synergistic or non-synergistic effect with the Lactobacillus crispatus strain according to the invention, or cosmetic agents with complementary effects.

[0115] These may include, for example, depigmenting and / or brightening ingredients and / or ingredients intended to reduce skin pigment spots, such as niacinamide or vitamin B3, arbutin, azelaic acid, ascorbic acid or its derivatives, a combination of plant extracts Saxifraga sarmentosa, Psidium guajava and Carica papaya, marketed by BASF Beauty Care Solutions France under the name Dermawhite™ WF, a combination of sulfites and extracts of Camellia sinensis, Scutellaria baicalensis, Cucumis sativus, Pyrus malus, marketed under the name Phytolight™ BG, a standardized extract of Lansium domesticum leaves marketed under the name DN-Aura™, a combination of a pea extract and sucrose dilaurate marketed under the name Actiwhite™, derivatives of acid 4-Hydroxyphenoxy acetic acid, in particular 2-(4-hydroxyphenoxy)-propionic acid marketed under the name Radianskin™ by BASF Beauty Care Solutions France,or a purified polysaccharide extract from Cassia angustifolia seeds marketed under the name Hyalurosmooth™, a stabilized protease extract isolated from Carica papaya latex marketed under the name X-Pressin™, a cationic liposome encapsulating ferulic acid marketed under the name Cytovector ferulic™, or a glycolic acid and L-arginine complex marketed, under the name AH-care™ by BASF Beauty Care Solutions France.

[0116] Among other active ingredients that may be combined with the strain according to the invention, these may include plant extracts possessing similar properties of preventing the appearance of pigmentation and / or increasing skin radiance, in particular an extract of the mushroom Inonotus obliquus marketed under the name Inolixir™ by the Applicant, an extract of Argania spinosa oil marketed under the name Arganyl™ by the Applicant, an extract of Moringa oleifera seeds marketed under the name Purisoft™ by the Applicant, a combination of an extract of Salvia miltiorrhiza and niacinamide marketed under the name CollRepair™ by the Applicant, an extract of Achillea millefolium marketed under the name Neurobiox™ by the Applicant, an extract of Cassia alata leaves marketed under the name DN-Age™ and / or an extract of lychee marketed under the Litchiderm™ is known for its antioxidant properties.of a chicory extract marketed under the name Lox-Age™, a yeast extract marketed under the name Vitacell™, a Polygonum bistorta extract marketed under the name Perlaura™, a galangal extract marketed under the name Hyalufix™, a corn extract marketed under the name Deliner™ or a Voandzeia subterranea extract marketed under the name Epigenist™ by the applicant or even active ingredients promoting skin firmness such as a synthetic tetrapeptide marketed under the name Dermican™, a Hibiscus abelmoschus extract marketed under the name Linefactor™, a purified pea extract marketed under the name Proteasyl™,An extract of Manilkara multinervis marketed under the name Elestan™ by the applicant. An extract of the plant Origanum majorana marketed under the name Dermagenist™ and / or an extract of Khaya senegalensis marketed under the name Collalift®18 and / or an extract of Eperua falcata marketed under the name Eperuline™ or an extract comprising phytosterols, in particular 3-sitosterol and / or campesterol and / or brassicasterol from rapeseed oil and marketed by the applicant under the name Phytosoothe™, may also be added to the cosmetic composition.

[0117] These may also include anti-aging active ingredients and / or tightening agents for a synergistic effect with the Lactobacillus crispatus strain. Advantageously, these include active ingredients that increase the gene and / or protein expression of collagen or active ingredients that prevent collagen degradation, such as retinol, vitamin C, an extract of Davilla rugosa marketed under the name Collguard™ by BASF Beauty Care Solutions, an extract of Hibiscus abelmoschus seed marketed under the name Linefactor™, a soy protein hydrolysate extract marketed under the name Phytokine™, a peptide marketed under the name Dermican™ by the applicant, or products marketed under the names Matrixyl™, Matrixyl 3000™ and Regestril™ by Sederma or Neuroguard by Codif, or Etemaline™ by Silab.

[0118] The tensor agents usable in the invention can be chosen from synthetic polymers, such as polyurethane latex or acrylic latex, polymers of natural origin, in particular polyholosides in the form of starch or in the form of carrageenans, alginates, agars, gellans, cellulosic polymers and pectins; plant proteins and protein hydrolysates; mixed silicates; wax microparticles; colloidal particles of inorganic filler chosen for example from silica, silica-alumina composites; as well as mixtures thereof.

[0119] Advantageously, the Lactobacillus crispatus strain according to the invention can be combined with a cosmetic agent selected from benzoic acid and / or its salts, levulinic acid and / or its salts such as sodium levulinate, sorbic acid and / or its salts, anisic acid and / or its salts, and mixtures thereof.

[0120] These may also include cosmetic ingredients such as antimicrobial agents, free radical scavengers, soothing, calming or relaxing agents, agents that act on microcirculation to improve skin radiance, particularly on the face, healing agents or slimming agents. Advantageously, the cosmetic and / or dermatological composition of the present invention also contains one or more tightening agents and / or one or more antimicrobial agents and / or one or more free radical scavengers and / or one or more soothing agents and / or one or more slimming agents and / or one or more agents active on microcirculation.

[0121] Among the antimicrobial agents associated with the non-viable form of the Lactobacillus crispatus strain in a preferred embodiment of the present invention, mention may be made of 2,4,4'-trichloro-2'-hydroxy diphenyl ether (or triclosan), 3,4,4'-trichlorobanilide, phenoxyethanol, phenoxypropanol, phenoxyisopropanol, hexamidine isethionate, metronidazole and its salts, miconazole and its salts, itraconazole, terconazole, econazole, ketoconazole, saperconazole, fluconazole, clotrimazole, butoconazole, oxiconazole, sulfaconazole, sulconazole, terbinafine, undecylenic acid and its salts, benzoyl peroxide, 3-Hydroxybenzoic acid, 4-Hydroxybenzoic acid, phytic acid, N-acetyl-L-cysteine, lipoic acid, azelaic acid and its salts, arachidonic acid, resorcinol, octoxyglycerin, octanoylglycine, caprylyl glycol, 10-hydroxy-2-decanoic acid, farnesol,Phytosphingosines and their mixtures.

[0122] Antioxidant agents may include vitamin C and its derivatives, including ascorbyl glucoside; phenols and polyphenols, in particular tannins, ellagic acid and tannic acid; epigallocatechin and natural extracts containing it, in particular green tea extracts; anthocyanins; phenolic acids; stilbenes; active ingredients that trap mono- or polycyclic aromatic compounds, tannins such as ellagic acid and indole derivatives and / or active ingredients that trap heavy metals such as EDTA, anti-radical active ingredients such as vitamin E and its derivatives such as tocopheryl acetate; bioflavonoids; coenzyme Q10 or ubiquinone.

[0123] As soothing agents included in the composition of the invention, pentacyclic triterpenes, ursolic acid and its salts, oleanolic acid and its salts, betulinic acid and its salts, salts of salicylic acid and in particular zinc salicylate, bisabolol, allantoin, omega-3 unsaturated oils, cortisone, hydrocortisone, indomethacin and beta methasone, anti-inflammatory agents, and in particular those described in application FR2847267, in particular Pueraria lobata root extract marketed under the name Inhipase™ by the applicant, Theobroma cacao extracts may be used.

[0124] The active ingredients acting on microcirculation, vasoprotectors or vasodilators, can be chosen from flavonoids, ruscogenins, nicotinates, essential oils.

[0125] Slimming agents may be chosen in particular from lipoprotein lipase inhibitors such as those described in US patent 2003086949 (Coletica) and in particular an extract of Peruvian liana (Uncaria tomentosa); draining agents, in particular hesperitine laurate (Flavagrum™), or quercetin caprylate (Flavenger™); phosphodiesterase enzyme inhibitors, adenylate cyclase activators, cAMP and / or agents capable of trapping spermine and / or spermidine. Examples include Coleus Forskohlii root extract, Cecropia obtusa extract, Uva lactuca extract, caffeine, forskolin, theophylline, theobromine and / or their derivatives, a hydrolyzed kappa carrageenan product called Slhnexcess™ marketed by the applicant and / or mixtures thereof.

[0126] The present invention also relates to a cosmetic treatment method comprising the topical application to at least one area of ​​healthy skin and / or healthy mucous membrane of a strain of Lactobacillus crispatus according to the invention, or of a cosmetic composition comprising it, to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or healthy mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on healthy skin and / or healthy mucous membranes and / or detoxify the skin and / or mucous membranes.

[0127] Advantageously, the Lactobacillus crispatus strain according to the invention, preferably in the form of a cosmetic composition according to the invention, is used in regular topical application and preferably at least once per day, advantageously twice a day, for at least 10 days, preferably for 20 days, and even more preferably for at least 28 days.

[0128] Advantageously, the topical application of a strain of Lacto-bacillus crispatus according to the invention or of a cosmetic composition comprising it, is carried out on the healthy skin and / or healthy mucous membranes of all or part of the body and / or face and / or scalp, preferably the legs, thighs, arms, belly, décolleté, neck, armpits, lips, again preferably all or part of the face, and preferably the cheeks, forehead, chin, lips, eye contour.

[0129] In an advantageous embodiment of the cosmetic process according to the invention, the cosmetic composition comprises the Lactobacillus crispatus strain according to the invention at a concentration of between 1x10⁴% and 10% (w / w) by weight, preferably between 1x10⁴% and 5% (w / w) by weight, further advantageously between 1x10³% and 0.5% (w / w) by weight, relative to the total weight of the composition, said composition further comprising a cosmetically acceptable excipient.

[0130] The cosmetic treatment process according to the invention aims to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on healthy skin and / or mucous membranes, advantageously comprises the following steps:

[0131] - The identification on the individual of an area of ​​healthy skin and / or healthy mucous membrane, for which one wishes to reduce the pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or mucous membranes, and / or to prevent the appearance and / or reduce the presence of pigment spots on healthy skin and / or mucous membranes, and

[0132] - Topical application to this area of ​​healthy skin and / or healthy mucous membrane, of a cosmetic composition comprising the Lactobacillus crispatus strain according to the invention, in an amount effective for reducing pigmentation of the skin and / or mucous membranes and / or preventing its increase, in particular for preserving and / or improving the complexion of healthy skin and / or mucous membranes, preferably for preserving and / or improving the radiance and / or luminosity and / or evenness of the complexion of healthy skin and / or mucous membranes, and / or for preventing the appearance and / or reducing the presence of pigment spots on healthy skin and / or mucous membranes healthy, namely in a Lactobacillus crispatus strain content advantageously between 1x104% and 10% (w / w) by weight, preferably between 1x104% and 5% (w / w) by weight, even more advantageously between 1x103% and 0.5% (w / w) by weight, relative to the total weight of the composition.

[0133] For the purposes of the present invention, "topically and / or orally acceptable" means an ingredient suitable for application by topical and / or oral route respectively, non-toxic, non-irritating to the skin including the scalp and not inducing an allergic response, and which is not chemically unstable.

[0134] The invention also relates to the Lactobacillus crispatus strain according to the invention, alone or in a pharmaceutical composition, preferably dermatological, comprising it, for its use, advantageously by topical route for the treatment and / or prevention of pathologies of the skin and / or mucous membranes involving hyperpigmentation and / or an increase in pigmentation, in particular a decrease in the radiance and / or luminosity and / or homogeneity of the complexion of the pathological skin and / or pathological mucous membranes, and / or an increase in the appearance of pigment spots of the pathological skin and / or pathological mucous membranes, and / or an increase in the activity of melanogenesis, in particular by way of an increase in melanin and / or tyrosinase activity, and / or an increase in oxidative stress of the pathological skin and / or pathological mucous membranes.

[0135] In particular, the invention relates to the Lactobacillus crispatus strain according to the invention or a pharmaceutical composition, preferably dermatological comprising it, for its use, advantageously by topical route, for the treatment and / or prevention of skin and / or mucous membrane pathologies associated with an increase in pigmentation of the skin and / or mucous membranes, in particular to depigment and / or prevent hyperpigmentation of pathological skin and / or pathological mucous membranes, and / or for the treatment and / or prevention of skin and / or mucous membrane pathologies associated with oxidative stress.

[0136] For the purposes of the present invention, from a dermatological point of view, "treatment and / or prevention of skin and / or mucous membrane pathologies to depigment and / or prevent hyperpigmentation" means a pigmentary disorder under the effect of skin and / or mucous membrane pathologies, such as Addison's disease, liver failure, purpura, melanoma, chloasma, dermatosis papulosa nigra, which is described as pathological hyperpigmentation, or even acne, eczema, atopic dermatitis, rosacea, couperose, redness.

[0137] For the purposes of the present invention, from a dermatological point of view, "treatment and / or prevention of pathologies of the skin and / or associated mucous membranes" means to oxidative stress”, the treatment and / or prevention of many skin diseases linked to excessive oxidative stress, including psoriasis, certain eczemas, acne, vitiligo, or even certain skin cancers.

[0138] The Lactobacillus crispatus strain according to the invention may be in the form of a pharmaceutical composition, preferably dermatological, comprising at least one pharmaceutically and / or dermatologically acceptable excipient. In one embodiment of the invention, said composition is applied topically and / or orally, preferably topically.

[0139] Advantageously, it is present in the pharmaceutical composition, preferably dermatological, at a concentration of 1x10 4% to 10% (w / w) by weight, preferably between 1x10 4% and 5% (w / w) by weight, further advantageously between 1x10 3% and 0.5% (w / w) by weight relative to the total weight of the composition, said composition further comprising at least one pharmaceutically, preferably dermatologically acceptable excipient.

[0140] The invention will be better understood upon reading the description of the figures and examples that follow.

[0141] Examples referring to the description of the invention are given below. These examples are provided by way of illustration and shall in no way limit the scope of the invention. Each example is general in scope.

[0142] The examples form an integral part of the present invention and any feature which appears new in relation to any prior art from the description taken as a whole, including the examples, forms an integral part of the invention.

[0143] On the other hand, in the examples, all percentages are given by weight unless otherwise stated, temperature is expressed in degrees Celsius unless otherwise stated, and pressure is atmospheric pressure unless otherwise stated.

[0144] Examples

[0145] Example 1. Preparation of different forms of Lactobacillus crispatus

[0146] Example La: production of viable whole form of Lactobacillus crispatus.

[0147] The isolated Lactobacillus crispatus (CNCM 1-5579) bacteria were inoculated into MRS (Man, Rogosa, Sharpe) agar and incubated anaerobically at 37°C. The pH was maintained at 6, and the medium was incubated for 24 hours. At the end of the incubation period, the medium was centrifuged, and the cell concentrate was lyophilized in the presence of maltodextrin, resulting in a final maltodextrin concentration of 50% (w / w) by weight relative to the total weight of the ingredient.

[0148] Example lb: production of the inactivated whole form of Lactobacillus crispatus

[0149] The isolated Lactobacillus crispatus (CNCM 1-5579) bacteria were inoculated in an MRS-type culture medium, incubated at 37°C under anaerobic conditions. The pH was The medium was maintained at a pH of 6 and incubated for 24 hours. At the end of the incubation period, the medium was centrifuged, and the cell concentrate was inactivated by heating it at 80°C for 30 minutes. The cell concentrate was then resuspended in glycerin.

[0150] Example Le: production of the lysate of the bacterium Lactobacillus crispatus

[0151] The isolated Lactobacillus crispatus (CNCM 1-5579) bacteria were inoculated The culture medium was incubated at 37°C under anaerobic conditions. The pH was maintained at 6, and the medium was incubated for 24 hours. At the end of the incubation period, the mixture was passed through a high-pressure homogenizer. The lysate was then resuspended in glycerin.

[0152] Example ld: Production of the secretome of the bacterium Lactobacillus crispatus

[0153] The isolated Lactobacillus crispatus bacteria (CNCM 1-5579) were inoculated into an MRS-type culture medium and incubated at 37°C under anaerobic conditions. The pH was maintained at pH 6, and the medium was incubated for 24 hours. At the end of the incubation period, the medium was filtered to recover the bacterial secretome. The secretome was then diluted in glycerin.

[0154] Example Le: production of the secretome of the bacterium Lactobacillus crispatus in powder form

[0155] Isolated Lactobacillus crispatus (CNCM 1-5579) bacteria were inoculated into MRS-type culture medium and incubated anaerobically at 37°C. The pH was maintained at 6, and the medium was incubated for 24 hours. At the end of the incubation period, the medium was filtered to recover the bacterial secretome. The secretome was then atomized with maltodextrin, resulting in a final maltodextrin concentration of 90% (w / w) by weight relative to the total weight of the ingredient.

[0156] Example 2: Inhibition of melanin synthesis by melanocytes in the presence of the Lactobacillus crispatus strain according to the invention.

[0157] Melanocytes from a murine B-16 cell line were seeded in 96-well plates at a rate of 8000 cells / cm2, then cultured in MEM (Minimum Essential Medium) supplemented with 10% FBS (Fetal Bovine Serum) at 37°C, 5% CO2 and 95% Relative Humidity for 3 days.

[0158] The medium was then removed and replaced with 200qL of the previously described medium with the addition of IqM of NDP-a-MSH (melanotropic hormone analogue) used as a stimulator of melanin synthesis and in the presence of the Lactobacillus crispatus strain according to example La at a concentration of 0.1% (w / w), for 5 days.

[0159] The same culture medium without the addition of the strain according to the invention but with the addition of IpM of NDP-a-MSH or with the addition of 0.01% (w / w) kojic acid were used as manipulation controls, respectively as a melanin-stimulating control and a melanin-inhibiting control.

[0160] The same culture medium without the addition of the strain according to the invention but with ethanol was used as a control (Control).

[0161] After 5 days of incubation, the cells were lysed with a solution of sodium hydroxide (NaOH) or potassium hydroxide (KOH). The absorbance was measured at 475 nm.

[0162] The amount of melanin was deduced from a standard curve consisting of different concentrations of commercial synthetic melanin.

[0163] The absorbance results were normalized with respect to the absorbance obtained with the same cell culture medium in the absence of the Lactobacillus crispatus strain according to the invention (Control). The results presented correspond to the average of 6 replicates (n=6).

[0164] Results

[0165] [Tables 1] Control (ethanol) NDP-α-MSH IpM (Melanin simulation control) Kojic acid 0.01% (w / w) (Melanin inhibition control) Product of the invention Lactobacillus crispatus (Example La at 0.1% w / w) Mean melanin (%) 24 100 18 48 Standard deviation 4 8 4 5 Statistics / NDP-α-MSH (*) P<0.001 (Student T test) NA (*) P<0.001 (Student T test) (*) p<0.05 (One-way ANOVA)

[0166] The Lactobacillus crispatus strain according to the example shown in the product of the invention demonstrated a significant 52% inhibition of melanin synthesis by murine B-16 melanocytes cultured in vitro. It can therefore be used to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase.

[0167] Example 3: Inhibition of tyrosinase activity in the presence of the Lactobacillus crispatus strain according to the invention.

[0168] Tyrosinase is a key enzyme in the process of melanin synthesis, which is responsible for skin tone. The tyrosinase used is extracted from normal human melanocytes.

[0169] The Lactobacillus crispatus strain according to example Le at a concentration of 0.05% (w / w) or a reference compound (kojic acid) at a concentration ranging from 0.25 mM to 8 mM, were pre-incubated with the enzyme for 10 minutes on ice. Next, the tyrosinase substrate, L-Tyrosine at a final concentration of 1 mM, was added to the medium and the plates were incubated at 37°C for 24 hours.

[0170] The same enzymatic extract without the addition of the strain according to the invention but with ethanol was used as a control (Control).

[0171] Enzymatic activity was evaluated by measuring melanin production by optical density (OD) at 540 nm using a microplate reader.

[0172] Optical density measurements were also carried out without the addition of substrate (n=l) in order to detect any possible interference from the Lactobacillus crispatus strain according to example Le or from the reference compound for each concentration tested.

[0173] The percentage of inhibition of tyrosinase activity was calculated according to the following formula:

[0174] % inhibition: 100-[DO value / Average DO of control x 100]

[0175] All experimental conditions were carried out in n=3. Intergroup comparisons were performed using an unpaired Student's t-test.

[0176] [Tables2] Untreated control (ethanol) Product of the invention Lactobacillus crispatus (Example Le at 0.05% (w / w)) Mean human tyrosinase inhibition (%) 0 99 Deviation from mean 3 0 Statistic / control NA *** (p<0.001)

[0177] The Lactobacillus crispatus strain according to Example Le of the invention has shown significant inhibition of tyrosinase activity extracted from human melanocytes by 99%. Through this inhibition of tyrosinase activity, the Lactobacillus crispatus strain can therefore be used to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase.

[0178] Example 4: Inhibition of melanin synthesis in a co-culture of melanocytes and keratinocytes in the presence of the Lactobacillus crispatus strain according to the invention.

[0179] Normal human keratinocytes and newborn human melanocytes were seeded at 20,000 cells / cm2 and 10,000 cells / cm2 respectively, then cultured for 7 days at 37°C under 5% CO2 and 95% relative humidity in a 50% / 50% (v / v) mixture of melanocyte culture medium and keratinocyte culture medium, supplemented with growth factors.

[0180] After one week of culture, the culture medium was replaced with a mixture of DMEM (Dulbecco Modified Eagle Essential Minimum Medium) culture medium without antibiotic and a melanocyte growth factor-free medium, and supplemented with 40pM oleic acid.

[0181] This culture medium without the addition of the strain according to the invention was used as an untreated control (Control).

[0182] To this same medium, the secretome of the bacterium Lactobacillus crispatus obtained according to example Le was added at a concentration of 0.1% (w / w) relative to the total weight of the culture medium and the secretome.

[0183] The same culture medium without the strain, but in the presence of Rucinol at 0.002% (w / w) and was used as a positive control.

[0184] The cultures were continued for 72 h at 37°C under 5% CO2 and 95% relative humidity, and then the culture medium was renewed. After an additional 48 h of culture, the medium was discarded, and the cells were rinsed with PBS (phosphate buffer). Then, a sodium hydroxide (SOH) solution containing 10% dimethyl sulfoxide (DMSO) was added to each well of the plate. 200 pL from each well were transferred to a transparent plate. The optical density, reflecting the amount of melanin, was read at 475 nm.

[0185] The results were the mean of the trials performed ± the standard deviation, expressed as a percentage, relative to the untreated control normalized to 100%. Statistical analysis of the results was performed relative to the untreated control using the One-Way ANOVA test.

[0186] Results

[0187] [Tables3] Untreated control Rucinol 0.002% (w / w) (Positive control) Product of the invention L-Actobacillus crispatus at 0.1% (w / w) (Example Le) Mean amount of melanin (%) 100 72 68 Standard deviation 10 3 7% inhibition NA 28 32 Statistics / control NA PcO.Ol PcO.Ol

[0188] The Lactobacillus crispatus strain according to Example Le of the invention showed a significant 32% reduction in melanin synthesis by melanocytes co-cultured in vitro with human dermal keratinocytes. Thus, the Lactobacillus crispatus strain, and in particular its secretome, can be used to reduce pigmentation of the skin and / or mucous membranes and / or prevent its increase.

[0189] Example 5: Reduction of oxidative stress in the presence of the Lactobacillus crispatus strain according to the invention.

[0190] Example 5.a: Decrease in the formation of DPPH° free radicals in tubo en presence of the Lactobacillus crispatus strain according to the invention.

[0191] The assay of the antiradical activity of the Lactobacillus crispatus strain was evaluated by a test using DPPH° (2,2-diphenyl-l-picrylhydrazyl).

[0192] The Lactobacillus crispatus strain according to Example La was mixed in PBS (Phosphate Buffer Saline) at a concentration of 0.05% (w / w), and according to Example Le at a concentration of 0.7% (w / w).

[0193] A negative control was made using the solvent for dissolving the strain, PBS, and a positive control was made with 1 mM vitamin C.

[0194] Each of the solutions containing the invention, the negative control or the positive control was mixed 50% / 50% (v / v) in a DPPH° solution, in 96-well plate wells.

[0195] Each of the solutions containing the invention, the negative control or the positive control was also mixed 50% / 50% (v / v) in an ethanol solution which is the solvent of DPPH°, in order to produce a blank, in the same plate.

[0196] The assembly was incubated for 30 minutes with shaking. At the end of incubation, the optical density (OD) was measured at a wavelength of 530 nm to estimate the amount of free radicals formed in each of the tested conditions.

[0197] The results of the DPPH° radical measurement to obtain the antiradical activity of the invention were the average of 6 trials (n=6) and were expressed as a mean percentage ± one standard deviation. The product of the invention was compared to its negative control (PBS alone in the absence of the Lactobacillus crispatus strain). The positive control, vitamin C, was compared to its negative control, water. Statistical analysis was performed using a one-way ANOVA for the invention and a Student's t-test for vitamin C.

[0198] [Tables4] Negative control (PBS) Vitamin C ImM Product of the invention (example at 0.7% (w / w)) Product of the invention Lactobacillus crispatus (Example at 0.05% (w / w)) Mean radicals (%) 100 0 68.81 66.19 Standard deviation 4.33 0 3.44 2.21 Statistic / control NA P<0.001 (Student's t test) Not tested P<0.001 (One-way ANOVA)

[0199] The Lactobacillus crispatus strain according to Example Ia of the invention showed significant in tubo free radical scavenging activity of 34%. The Lactobacillus crispatus strain according to Example Ia of the invention showed in tubo free radical scavenging activity of 31%. By reducing the production of free radicals, the Lactobacillus crispatus strain can therefore be used to detoxify the skin and / or mucous membranes, to reduce pigmentation of the skin and / or mucous membranes, and / or prevent its increase.

[0200] Example 5.b: Reduction in the release of cellular free radicals induced by UVs within fibroblasts in the presence of the Lactobacillus crispatus strain according to the invention.

[0201] Human fibroblasts described as "normal", i.e., not exhibiting any pathology, from a healthy donor were seeded in 24-well plates at a rate of 30,000 cells / cm2, then cultured in a DMEM (modified Eagle minimum essential medium) / Ham's F-12 medium, supplemented with 10% FBS and 0.5% antibiotic, for 96h at 37°C, under 5% CO2 and 95% relative humidity.

[0202] Then, 0.5 mL of the cell suspension was incubated for 72 h in the presence of the strain Lactobacillus crispatus according to example la at a concentration of 0.0125% (w / w), or of an antioxidant control (Vitamin E at 0.0003% (v / v)) in an EMEM medium (Eagle's minimum essential medium) supplemented with 1% S VF and 0.5% antibiotic.

[0203] This culture medium without the addition of the strain according to the invention was used as an untreated control (UVA only control - Oxidative stress control).

[0204] A DCFH-DA (2',7'-dichlorodihydrofluorescein diacetate) probe solubilized at 100 µM in PBS (phosphate buffer) was incubated with the cells. After the first hour of incubation at 37°C under 5% CO2 and 95% relative humidity, the cells were rinsed with PBS and irradiated with UVA at 20 J / cm² for 3 hours. An unirradiated control was also prepared. After irradiation, the irradiated medium was discarded, and the cells were rinsed with PBS.

[0205] The fluorescence reading on the cell mat related to the formation of DCF (2',7-dichlorofluorescein) from the probe (DCFH-DA) was read at 485 nm excitation and 538 nm emission.

[0206] The results were the mean of 12 replicates (n=12) and were expressed as mean ± one standard deviation. The results were compared to the UVA-only control normalized to 100% and statistically compared using the one-way ANOVA test.

[0207] [Tables5] UVA control only (Oxidative stress control) Vitamin E 0.0003% (v / v) Product of the invention Lactobacillus crispatus (Example at 0.0125% (w / w)) Mean radicals measured by DCF fluorescent probe (%) 100 71 44 Standard deviation 8 25 21 Statistic / control NA P<0.05 (Student's t-test) P<0.05 (Student's t-test)

[0208] The Lactobacillus crispatus strain according to Example La of the invention has shown a significant 56% reduction at the cellular level in the release of intracellular free radicals induced by UVA oxidative stress. By reducing the release of intracellular free radicals induced by UVA, the Lactobacillus crispatus strain can therefore be used to detoxify the skin and / or mucous membranes and / or preserve and / or improve the radiance and / or luminosity and / or evenness of the complexion. skin and / or mucous membranes, preferably to reduce the presence and / or prevent the appearance of pigment spots on the skin and / or mucous membranes, particularly when these are subjected to oxidative stress, preferably by UVA.

[0209] Example 5.c: Reduction of UV-induced membrane lipid peroxidation within fibroblasts in the presence of the Lactobacillus crispatus strain according to the invention.

[0210] Human fibroblasts described as "normal", i.e., not exhibiting any pathology, from a healthy donor were seeded in 24-well plates at a rate of 30,000 cells / cm2, then cultured in a DMEM / Ham's F-12 medium, supplemented with 10% FBS and 0.5% antibiotic, for 96h at 37°C, under 5% CO2 and 95% relative humidity.

[0211] Then, 0.5 mL of the cell suspension was incubated for 72 h in the presence of the Lactobacillus crispatus strain according to example 1a at a concentration of 0.00625% (w / w), or of an antioxidant control (Vitamin E at 0.0003% (v / v)) in an EMEM medium supplemented with 1% FBS and 0.5% antibiotic.

[0212] This culture medium without the addition of the strain according to the invention was used as an untreated control (after irradiation, constitutes the UVA only control - Oxidative stress control).

[0213] After the first hour of incubation at 37°C under 5% CO2 and 95% relative humidity, the cells were rinsed with PBS and irradiated with UVA at 20 J / cm² for 3 hours. A non-irradiated control was also prepared. After irradiation, the supernatants were collected and the cells were rinsed with PBS.

[0214] A 0.5 mL aliquot of the supernatants was transferred to a Pyrex glass tube to which a 40% trichloroacetic acid and 2% thiobarbituric acid solution was added. The resulting mixture was heated for 30 minutes at 100°C. Then, the formation of MDA (MalonDiAldehde), an oxidation product released following UVA-induced lipid peroxidation, was measured by fluorescence at 532 nm excitation and 560 nm emission.

[0215] The results were the mean of the trials performed and were expressed as mean ± one standard deviation. The results were compared to the UVA-only control normalized to 100% and statistically compared using the one-way ANOVA test.

[0216] [Tableauxô] UVA control alone (Oxidative stress control) Vitamin E 0.0003% (v / v) Product of the invention Lactobacillus crispatus (Example at 0.00625% w / w) Mean MDA released (%) 100 48 68 Standard deviation 11 10 12 Statistic / control NA P<0.001 (Student's t-test) P<0.001 (Student's t-test)

[0217] The Lactobacillus crispatus strain according to Example La of the invention has shown a significant 32% reduction at the cellular level in the formation of UVA-induced MDA. By reducing lipid peroxidation in the presence of UV stress, the Lactobacillus crispatus strain can therefore be used to detoxify the skin and / or mucous membranes, particularly when they are subjected to oxidative stress, especially UVA stress.

[0218] Example 6: Stimulation of cellular antioxidant defenses after UV irradiation within fibroblasts in the presence of the Lactobacillus crispatus strain according to the invention.

[0219] Human fibroblasts described as "normal", i.e., not exhibiting any pathology, from a healthy donor were seeded in 24-well plates at a rate of 30,000 cells / cm2, then cultured in a DMEM / Ham's F-12 medium, supplemented with 10% S VF and 0.5% antibiotic, for 96h at 37 °C, under 5% CO2 and 95% relative humidity.

[0220] This culture medium without the addition of the strain according to the invention was used as an untreated control (after irradiation, constitutes the UVA only control - Oxidative stress control).

[0221] Then, 0.5 mL of the cell suspension was incubated for 72 h with the Lactobacillus crispatus strain according to example La at a concentration of 0.0125% (w / w), or with an antioxidant control (Vitamin E at 0.0003% (v / v)) in an EMEM medium supplemented with 1% S VF and 0.5% antibiotic.

[0222] After the first hour of incubation at 37°C under 5% CO2 and 95% relative humidity, the cells were rinsed with PBS and irradiated with UVA at 20 J / cm² for 3 hours. A non-irradiated control was also prepared. After irradiation, the irradiated medium was removed, and the cells were rinsed with PBS.

[0223] Sodium hydroxide (NaOH)₂IN was added to each cupule, followed by a solution of o-Phthaldialdehyde (OPT) diluted 1 / 15 in GSH buffer (reduced L-glutathione). The mixture was incubated for 15 minutes at room temperature. The amount of glutathione was then measured by fluorescence at 355 nm excitation and 430 nm emission.

[0224] The results were the average of the tests performed and were expressed as a mean. + / - one standard deviation. The results were compared to the UVA-only control normalized to 100% and statistically compared using the One Way ANOVA test.

[0225] [Tables?] UVA control only (Oxidative stress control) Vitamin E 0.0003% (v / v) Product of the invention Lactobacillus crispatus (Example at 0.0125% (w / w)) Mean glutathione (%) 100 100 154 Standard deviation 11 13 41 Statistic / control NA NS P<0.05 (Student's t-test)

[0226] The Lactobacillus crispatus strain according to Example La of the invention showed a significant increase in cellular antioxidant defenses by an increase in glutathione of 54% after UVA irradiation.

[0227] By this action of increasing cellular antioxidant defenses in the presence of oxidative stress by UVAs, the Lactobacillus crispatus strain can therefore be used to detoxify the skin and / or mucous membranes, in particular when they are subjected to oxidative stress, in particular stress by UVAs.

[0228] Example 7: In vivo measurement of the decrease in the amount of melanin in the skin in the presence of the Lactobacillus crispatus strain according to the invention.

[0229] A population of 30 Chinese women of phototype II to IV, aged 40 to 50 years with pigment spots on the face applied to half of their face a cream in the form of an emulsion comprising a final concentration by weight of the Lactobacillus crispatus strain according to example Le of 1% (w / w) relative to the total weight of the cream (Formulation example 8.b), or without said extract replaced by water (placebo), at a rate of 2 applications per day for 2 months.

[0230] The evaluation of the skin's melanin content was carried out using a device called a Mexameter, which uses the principle of absorption / reflection. From the Mexameter measurements, a melanin index is deduced. A decrease in this index corresponds to a decrease in melanin.

[0231] The efficacy of the composition containing the Lactobacillus crispatus strain according to the example was compared with that of the so-called placebo emulsion.

[0232] The melanin index was measured at the beginning of the study (DO) after one month of application (D28) and finally after 2 months of application (D56).

[0233] The efficacy of the formulation containing the Lactobacillus crispatus strain according to example Le and of the placebo formula were expressed as mean and were compared with the baseline values ​​(DO) and compared between the formulation containing the Lactobacillus crispatus strain with that containing the placebo, using the Student's t-test or the Wilcoxon test.

[0234] [Tables8] Product of the invention: Lactobacillus crispatus (Example at 1% (w / w)) Placebo Melanin index at D28 (in % vs D0) -3.1% (p<0.001 vs D0; p<0.001 vs placebo) -0.1% Melanin index at D56 (in % vs D0) -5.5% (p<0.01 vs D0; p<0.01 vs placebo) +1%

[0235] The formulation of the Lactobacillus crispatus strain according to example Le significantly decreased the amount of cutaneous melanin measured via the melanin index using a Mexameter.

[0236] These results show the beneficial effect of the Lactobacillus crispatus strain formulated to preserve and / or improve the radiance and / or luminosity and / or homogeneity of healthy skin and / or healthy mucous membranes.

[0237] Example 8: Example of a cosmetic composition comprising the Lactobacillus crispatus strain according to the invention

[0238] Example 8.a: Example of a cosmetic composition comprising the Lactobacillus crispatus strain according to example La

[0239] The Lactobacillus crispatus strain obtained according to Example La was added just before application to the skin in a formulation as defined below according to Table 8, at a final concentration of 0.05% (w / w) relative to the total weight of the formulation.

[0240] [Tables9] Product Name Quantity (% by total weight) Phase A Cetearyl alcohol, lecithin, sodium cetearyl sulfate, olive oil 3.00 Hydrogenated vegetable glycerides 2.50 Caprylyl caprylate / caprate 6.50 Dicaprylyl carbonate 4.00 Phase B Water 66.35 Butylene glycol 10.00 Sodium benzoate 0.25 Phase C Glycerin 5.00 Xanthan gum 1.00 Phase D Glycerin, water, sodium levulinate, sodium anisate 1.00 Citric acid 0.40

[0241] Example 8.b: Example of a cosmetic composition comprising the Lactobacillus crispatus strain according to example le

[0242] Proportions are expressed as % and names in capital letters correspond to the INCI names of the ingredients.

[0243] [TableauxlO] Phase A Glycerin 5.00 Butylene Glycol 10.00 Xanthan Gum 2.00 Phase B Water 58.95 Sodium Benzoate 0.25 Glycerin, Water, Sodium Levulinate, Sodium Anisate 1.00 Phase C Polyglyceryl-2 Dipolyhydroxystearate 1.00 Dipropylheptyl Carbonate 3.00 Dicaprylyl Carbonate 3.00 Caprylyl Caprylate / Caprate 10.00 Laureth-7 Citrate 0.50 Phase D Citric Acid 0.30 Phase E Lactobacillus crispatus Extract obtained according to example 1.00 Water 4.00

[0244] The different phases A, B, and C are mixed together at room temperature using methods known to those skilled in the art, by adding B to A while stirring, and then C to AB. The pH is adjusted to 4.9 with D. Then phase E is added. Finally, the mixture is homogenized using Ultra Turrax to prepare a composition according to the present invention.

Claims

Demands

1. Cosmetic and / or nutraceutical use of a strain of Lactobacillus crispatus, preferably the Lactobacillus crispatus strain being the species registered under the designation CNCM 1-5579, to decrease pigmentation of the skin and / or mucous membranes and / or prevent its increase.

2. Use according to claim 1, wherein the strain decreases and / or inhibits melanogenesis, in particular by acting on the decrease and / or inhibition of melanin, preferentially on the decrease and / or inhibition of the synthesis and / or quantity of melanin, and / or by decreasing and / or inhibiting the activity of tyrosinase, and / or by detoxifying healthy skin and / or healthy mucous membranes.

3. Use according to any of the preceding claims to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes.

4. Use according to any one of the preceding claims, to preserve and / or improve the radiance and / or luminosity and / or evenness of the complexion, of healthy skin and / or healthy mucous membranes.

5. Use according to any one of the preceding claims, to prevent the appearance and / or reduce the presence of pigment spots on healthy skin and / or healthy mucous membranes, preferably brown spots and / or age spots.

6. Use according to any one of the preceding claims, to detoxify healthy skin and / or healthy mucous membranes, in particular by an antioxidant action and / or on lipid peroxidation and / or by an increase and / or stimulation of glutathione synthesis induced by oxidative stress.

7. Use according to any one of the preceding claims, wherein the strain is used in whole form, in particular viable and / or inactivated, in particular dead, and / or in lysate form, and / or in the form of one or more of its fractions, and / or in the form of one or more of its metabolites, preferably its secretome.

8. Use according to any one of the preceding claims, wherein the Lactobacillus crispatus strain is that filed under number CNCM 1-5579.

9. Use according to any one of the preceding claims, wherein the Lactobacillus crispatus strain is used in viable whole form and / or in the form of one or more of its metabolites, preferably from its secretome.

10. Use according to any one of the preceding claims, wherein the Lactobacillus crispatus strain is associated with its culture medium.

11. Use according to any one of the preceding claims, wherein the Lactobacillus crispatus strain is applied topically to healthy skin and / or healthy mucous membranes.

12. Use according to claim 11 wherein the skin is phototype IV to VI.

13. Use according to any one of the preceding claims, such that the Lactobacillus crispatus strain is present in a cosmetic composition at a concentration of 1x104% to 10% by weight, preferably between 1x104% and 5% by weight, further advantageously between 1x103% and 0.5% by weight, relative to the total weight of the composition, said composition further comprising at least one cosmetically acceptable excipient.

14. A cosmetic treatment method characterized in that it comprises the topical application to at least one area of ​​healthy skin and / or healthy mucous membrane of a strain of Lactobacillus crispatus as defined according to any one of claims 7 to 10, or of a cosmetic composition according to claim 13, to decrease pigmentation of the skin and / or mucous membranes and / or prevent its increase, in particular to preserve and / or improve the complexion of healthy skin and / or healthy mucous membranes, preferably to preserve and / or improve the radiance and / or luminosity and / or homogeneity of the complexion of healthy skin and / or healthy mucous membranes, and / or to prevent the appearance and / or decrease the presence of pigment spots on healthy skin and / or healthy mucous membranes and / or detoxify the skin and / or mucous membranes.

15. A cosmetic treatment method according to claim 14, wherein the topical application of a strain of Lactobacillus crispatus as defined in any one of claims 7 to 10, or of a cosmetic composition according to claim 13, is carried out on healthy skin and / or healthy mucous membranes of all or part of the body and / or face and / or scalp, preferably the legs, thighs, arms, abdomen, décolleté, neck, armpits, lips, again preferably all or part of the face, and preferably the cheeks, forehead, chin, lips, eye contour.

16. Cosmetic treatment method according to any one of the claims 14 to 15, wherein the cosmetic composition comprises the Lactobacillus crispatus strain as defined according to any one of claims 7 to 10 at a concentration of 1x10⁴% to 10% by weight, preferably between 1x10⁴% and 5% by weight, further advantageously between 1x10⁳% and 0.5% by weight, relative to the total weight of the composition, said composition further comprising at least one cosmetically acceptable excipient.

17. A strain of Lactobacillus crispatus, or a pharmaceutical composition, preferably dermatological, comprising it, for use, advantageously by topical route, for the treatment and / or prevention of skin and / or mucous membrane diseases to depigment and / or prevent hyperpigmentation of pathological skin and / or mucous membranes, said pathology being selected from Addison's disease, liver failure, purpura, melanoma, chloasma, dermatosis papulosa nigra, acne, atopic dermatitis, rosacea, redness, and / or for the treatment and / or prevention of skin and / or mucous membrane diseases associated with oxidative stress, said pathology being vitiligo.

18. Strain according to claim 17 wherein the strain is used in whole form, in particular viable and / or inactivated, in particular dead, and / or in lysate form, and / or in the form of one or more of its fractions, and / or in the form of one or more of its metabolites, preferably its secretome, and / or wherein the strain is associated with its culture medium.

19. A strain of Lactobacillus crispatus for use according to any one of claims 17 to 18, characterized in that it is in the form of a pharmaceutical composition, preferably dermatological, comprising at least one dermatologically acceptable excipient.

20. A strain of Lactobacillus crispatus for use according to any one of claims 17 to 19, characterized in that it is present in the pharmaceutical composition, preferably dermatological, at a concentration of 1x10⁴% to 10% by weight, preferably between 1x10⁴% and 5% by weight, further advantageously between 1x10⁳% and 0.5% by weight, relative to the total weight of the composition, said composition further comprising at least one pharmaceutically, preferably dermatologically, acceptable excipient.

21. Lactobacillus crispatus strain for its use according to one any of claims 17 to 20, characterized in that the strain is as defined according to any one of claims 7 to 10.