WO2014096084A2 - Use of probiotic microorganisms as an agent for promoting melanin synthesis - Google Patents

Abstract

The invention relates to the oral use of Bifidobacterium longum as a pro-pigmenting agent, in particular in the prevention and/or treatment of alterations in the homogeneity of the colour of the skin and/or hair.

Description

 Use of probiotic microorganisms as an agent promoting the synthesis of melanin

The present invention relates to the cosmetic or therapeutic treatment, orally, by induction of pigmentation, alterations in the homogeneity of the color of the skin and / or the hair, resulting in particular from a regulation imbalance of the skin. homeostasis of melanocytes.

The color of human hair and skin is a function of various factors, such as seasons of the year, ethnicity, gender, and age. It is mainly determined by the concentration and distribution in keratinocytes of melanin produced by melanocytes. Melanin, a natural pigment known for its anti-radical and absorbing properties of solar radiation, is a physiological protective agent of the skin, existing in two major forms: eumelanin and pheo-melanin.

 Melanocytes are the specialized cells that synthesize melanin and then distribute it to keratinocytes through particular organelles, melanosomes.

 In the epidermis, the melanocyte is involved in the epidermal melanic unit, forming a distribution network between melanocytes and keratinocytes, which consists of a melanocyte surrounded by about 36 neighboring keratinocytes. Melanocytes account for approximately 5 to 10% of basal layer cells of the epidermis. All individuals, regardless of phototype, have approximately the same number of melanocytes for a given skin area. Differences in pigmentation between individuals are not due to the number of melanocytes, but to the nature of the synthesized melanin and the biochemical and functional properties of melanosomes.

Melanosomes are highly specialized organelles whose sole function is the synthesis and transfer of melanin. They arise from the endoplasmic reticulum in the form of spherical vacuoles called pre-melanosomes that contain an amorphous protein substrate, but no melanogenic enzymes. During the maturation of the pre-melanosome, the amorphous substrate is organized into a fibrillar structure oriented in the longitudinal axis of the melanosome. There are four stages of development of melanosome corresponding to the intensity of melanization. Melanin is uniformly deposited on the inner fibrillar network of the melanosome and the opacity of the organelle increases to saturation. As melanosomes are synthesized in melanosomes, they move from the perinuclear region to the end of the dendrites of melanocytes. By phagocytosis, the end of the dendrites is captured by the keratinocytes, the membranes are degraded and the melanosome content of the melanosomes is redistributed in the keratinocytes. The melanin is thus distributed in the epidermis, ensuring its browning and protection.

 In terms of hair and hair color, it is based on the presence in varying amounts and ratios of two groups of melanins: eumelanins (brown to black pigments) and pheo-melanins (red to yellow pigments). The pigmentation of hair and hair requires the presence of melanocytes in the bulb of the hair follicle. The hair follicle is a tubular invagination of the epidermis that penetrates to the deep layers of the dermis. The inner part of the bulb is a zone of cellular proliferation where the precursors of the keratinized cells constituting the hair are found. The melanocytes in the bulb of the hair follicle are in an active state, that is, they synthesize melanin. These pigments are transmitted to the keratinocytes intended to form the hair shaft, which will lead to the growth of a pigmented hair or hair.

The mechanisms leading to the pigmentation of the skin and keratinous fibers, especially hair and hair, are highly regulated mechanisms, under the influence of multiple hormonal or cellular factors.

 The homeostasis of melanocytes, finely regulated by these hormonal or cellular factors, makes it possible to confer on skin and keratin fibers a natural, homogeneous, more or less intense pigmentation, imparting an attractive aesthetic and cosmetic appearance.

 However, in most populations, obtaining a brown color of the skin and maintaining a constant color of the hair are important aspirations. The maintenance of a tanned complexion and a naturally pigmented hair are factors strongly involved in obtaining an attractive cosmetic appearance.

 In addition, changes in the physiology of the epidermis and hair follicle, particularly related to age, can alter the pigmentation of the skin, hair and hair. In particular, these modifications may result in dysregulation of melanocyte homeostasis, manifested by disorders of proliferation, maturation or survival, possibly accompanied by an abnormality or an imbalance of the melanogenesis.

On the other hand there are diseases of pigmentation such as vitiligo which is an autoimmune disease characterized by the appearance on the skin of white patches related to a lack of pigmentation. There is therefore a real need for a product that facilitates and / or improves the pigmentation of the skin and / or the hair and / or hair, and can be used orally.

 Numerous solutions have been proposed in the field of artificial coloring by adding exogenous dyes, such as DHA, which are supposed to give the skin and / or the hair and / or the hair the coloring as close as possible to what it is. is naturally or, in the field of natural staining, by stimulation of the natural pigmentation pathways, for example using active agents stimulating melanogenesis with or without UV action.

 It has thus been proposed to use compositions containing a phosphodiesterase inhibitor (WO 95/17161), prostaglandins (WO 95/1 1003), DNA fragments (WO 95/01773), DNA encoding MSH receptors (WO 94/04674), diacylglycerols (WO 94/04122), tyrosine derivatives (EP 0585018), ballot extracts (WO 93/10804), or xanthine extracts (WO 91/07945) . These different compounds act, directly or indirectly via Ι'α-MSH or prostaglandins, so as to stimulate the biosynthesis of melanin, with or without UV action.

 Excellent results are certainly obtained by the solutions proposed in the state of the art, but the compounds used often have significant side effects or are complex mixtures that do not have specificity.

 The discovery of alternative substances having an effect on the pigmentation of the skin and / or hair and / or hair therefore remains a major objective of the research.

 In particular, there is still a need for alternative assets, in particular cosmetics, which can preferably be administered orally, which physiologically induce pigmentation.

 There is still a need for alternative assets that promote melanogenesis.

 There is still a need for alternative active ingredients to regulate the synthesis or accumulation of melanin in the epidermis.

 There is still a need for alternative assets to increase melanin deposits in hair and / or hair.

 There is still a need for alternative assets to facilitate and / or improve the pigmentation of the skin and / or hair and / or hair in the field of dermatology and cosmetics.

There is still a need for alternative assets useful to prevent and / or limit and / or stop the development of canities, or even maintain the natural pigmentation of hair and / or gray or white hairs. The present invention is intended to meet these needs.

Probiotic microorganisms, microorganisms that can have a positive effect on the health of the subject who ingests them, are widely used in the field of cosmetics.

 Thus, application FR 2 756 181 describes the use of cosmetic compositions based on inactivated culture of bacteria of the Bifidobacterium type, mint oil and an acid to eliminate pigment spots.

 The application EP 1 1 10 555 describes the use of a bacterial agent, for example an extract of Bifidobacterium bacteria, to stabilize and / or regenerate the cutaneous ecosystem of mammals, and treat, inter alia, pityriasis versicolor.

 Application FR 2 912 917 describes the use of a conditioned culture medium, obtained by contact with peripheral blood cells stimulated by probiotics, to treat signs of inflammation and / or immune disorders, such as inflammatory hyperpigmentations, vitiligo or canitia.

 WO 2008/015343 describes the use of yeast extracts to increase melanin synthesis, and treat vitiligo and canitie.

 The application WO 2007/073122 describes the use of a fermented rice bran with a lactic acid bacteria to whiten the skin by preventing the synthesis of melanin.

 US 2002/0168388 discloses the use of compositions comprising inactivated bacteria and extracellular plant matrix extract to counter UV radiation-induced damage.

 The application WO 02/28402 describes the use of compositions comprising probiotics for balancing the immune function of the skin after exposure to stress conditions.

 The application FR 2 834 718 describes the use of active substances obtained by fermentation of vegetable seeds or fruits with Lactobacillus, Lactococcus or Leuconostoc bacteria to protect the skin from the harmful effects of UV rays and to regulate melanogenesis in the skin and skin. hair.

 The application FR 2 920 307 describes the use of probiotics to treat the manifestations of discomfort and / or cutaneous signs associated with a surface skin treatment, among which the bleaching of the skin.

The application FR 2 920 306 describes the use of a Bifidobacterium lysate to prevent a decrease in and / or enhance the skin barrier function, and thus prevent and / or reduce skin irritations or signs of skin aging. The application FR 2 920 300 describes the use of hesperidin in combination with a probiotic to enhance the barrier function of the skin and thus treat the cutaneous signs of aging.

 The use of probiotic Bifidobacterium as a pro-pigmenting agent, however, has not been suggested.

 However, the inventors have surprisingly shown that Bifidobacterium longum bacteria significantly induce cutaneous or capillary pigmentation. In particular, this specific probiotic strain makes it possible to induce a greater pigmentation than other types of known probiotics such as strains of Propionibacterium freudenreichii or Bacillus coagulans.

 The present invention therefore relates to the non-therapeutic cosmetic use of at least one strain of probiotic microorganism Bifidobacterium longum as agent inducing cutaneous and / or capillary pigmentation, to homogenize the color of the skin and / or hair, said at least one a strain of probiotic microorganism Bifidobacterium longum being administered orally and said at least one strain of Bifidobacterium longum not being the strain Bifidobacterium longum subsp. longum filed on 29 January 2001 with the CNCM (Paris, France) under the number 1-2618.

 It also relates to a strain of probiotic microorganism Bifidobacterium longum for use as an agent inducing cutaneous and / or capillary pigmentation, for the prevention and / or treatment of an alteration of the homogeneity of the skin color and / or hair of pathological origin, said strain of probiotic microorganism Bifidobacterium longum being administered orally and said strain of Bifidobacterium longum not being the strain Bifidobacterium longum subsp. longum filed on 29 January 2001 with the CNCM (Paris, France) under the number 1-2618.

Detailed description of the invention

Bifidobacterium longum

 The bacterial strain Bifidobacterium longum used in the context of the present invention is a probiotic microorganism.

 "Probiotic microorganism" means a living microorganism which, when administered in an adequate amount, has a positive effect on the health of its host

("Joint FAOAA HO Expert 10 Consultation on Evaluation of Health and Nutritional Properties of Probiotic in Food Including Powder Milk with Live Lactic Acid Bacteria, October 6, 2001"). For the purposes of the present invention, the microorganism envisaged may be used live, or inactivated, unable to replicate. It is also possible to use a fraction of it or one of the components of this microorganism.

 According to a variant of the invention, the probiotic microorganism Bifidobacterium longum used in the context of the invention may be used in an isolated or purified form, that is to say unmixed with one or more compound (s) likely to be associated with it in its environment of origin or in its environment of propagation.

 According to a variant of the invention, the probiotic microorganism Bifidobacterium longum used in the context of the invention can be implemented in a living, semi-active, inactivated or dead form.

 In particular, the probiotic microorganism Bifidobacterium longum used in the context of the invention can be used in a living or inactivated form.

 According to an advantageous embodiment of the invention, the probiotic microorganism Bifidobacterium longum can be used in inactivated or dead form.

 For the purposes of the invention, an "inactivated" microorganism is a microorganism that is no longer capable, temporarily or permanently, of forming colonies in culture.

 For the purposes of the invention, a "dead" microorganism is a microorganism that is no longer capable, definitively, of forming colonies in culture.

 Dead or inactivated microorganisms may have intact or broken cell membranes. Thus, the term "inactivated" also refers to extracts and lysates of microorganisms as detailed below. Obtaining dead or inactivated microorganisms can be carried out by any method known to those skilled in the art.

 According to an advantageous embodiment, the probiotic microorganisms Bifidobacterium longum used according to the invention are at least partly inactivated or dead.

"At least partially inactivated probiotic microorganisms Bifidobacterium longum" means a preparation of probiotic microorganisms Bifidobacterium longum according to the invention comprising at least 80%, in particular at least 85%, more particularly at least 90%, or at least 95%, at least 99%, or at least 99.99% of inactivated Bifidobacterium longum probiotic microorganisms expressed in colony forming units (cfu) relative to all of the non-inactivated live Bifidobacterium longum probiotic microorganisms contained in the initial preparation prior to be subjected to an inactivation process. The inactivation rate obtained depends on the conditions of application of the inactivation method which are adjusted by those skilled in the art according to the inactivation rate to be obtained. According to one embodiment, the invention comprises the use of a preparation comprising a maximum, preferably 100%, of inactivated Bifidobacterium longum probiotic microorganisms.

 An inactivated Bifidobacterium longum probiotic microorganism suitable for the invention may be prepared by irradiation, heat treatment or, under certain conditions, lyophilization of a preparation of probiotic microorganisms Bifidobacterium longum. These methods are known to those skilled in the art. Other known techniques among which high pressure treatment or extrusion can also be envisaged by the skilled person.

 In particular, the inactivation of probiotic microorganisms by irradiation may include the use of gamma rays, X-rays, or UV exposure. The type of radiation, the intensity, the dose and the exposure time are adjusted by those skilled in the art according to the quantity and the nature of the probiotic Bifidobacterium longum microorganisms to be inactivated.

 Thermal inactivation can be accomplished by incubating the probiotic Bifidobacterium longum microorganisms of the invention for a given period of time, for example from 10 s to 90 min at a temperature of 100 to 150 ° C. Depending on the microorganism to be inactivated, it is possible to envisage a longer treatment, for example 2 hours, at 170 ° C.

 Thermal inactivation can also be performed by autoclaving by subjecting the probiotic Bifidobacterium longum microorganisms of the invention to a temperature of 121 ° C, for at least 20 minutes and at an atmospheric pressure of 2 bar.

 Alternatively, the thermal inactivation can be carried out by subjecting the Bifidobacterium longum probiotic microorganisms to a freezing temperature, for a prolonged period of time, or to a series of freeze-thaw cycles.

 The inactivation by lyophilization can be carried out by any method known in the art. Advantageously, probiotic microorganisms Bifidobacterium longum inactivated by lyophilization can be returned to culture.

 Preferably, a probiotic microorganism Bifidobacterium longum suitable for the invention is used in an inactivated form obtained by irradiation, in particular by gamma irradiation.

A probiotic microorganism Bifidobacterium longum according to the invention may be used in whole form, that is to say substantially in its native form, or in the form of extracts or lysates comprising fractions and / or metabolites of this microorganism. For the purposes of the invention, the term "fraction" refers to a fragment or component of said microorganism having an induction efficiency of melanin synthesis by analogy with said whole microorganism.

 For the purposes of the invention, the term "metabolite" refers to any substance derived from the metabolism of microorganisms, and in particular secreted by the microorganisms Bifidobacterium longum considered according to the invention and also having an induction efficiency of melanin synthesis.

 For the purposes of the invention, the term "extract" means a preparation containing inactivated microorganisms whose cell structure has not been destroyed or dissolved. Biological cells are generally not fragmented.

 An extract or a lysate suitable for the invention may be prepared from Bifidobacterium longum bacteria at the end of the growth phase.

 According to a preferred embodiment, a probiotic microorganism Bifidobacterium longum suitable for the invention can be prepared in the form of a lysate.

 A lysate in the sense of the invention commonly refers to a material obtained at the end of the destruction or dissolution of biological cells by a so-called cell lysis phenomenon thus causing the release of the intracellular biological constituents naturally contained in the cells of the microorganism under consideration and of fragments of cell membrane components. For the purposes of the present invention, the term "lysate" is used interchangeably to designate the entirety of the lysate obtained by inactivation of the microorganism concerned or only a fraction thereof. The lysate used is therefore formed in all or part of the intracellular biological constituents and constituents of the cell walls and membranes of the probiotic microorganism Bifidobacterium longum of interest.

 It is advantageous to use in the context of the invention:

 an extract containing inactivated microorganisms of the relevant microorganism strain, or

 a lysate obtained by lysis or inactivation of the microorganism concerned. This cellular inactivation for obtaining an extract or a lysate can be accomplished by various technologies, such as, for example, osmotic shock, thermal shock, ultrasound, or else under mechanical stress of centrifugation type, or combinations of these different technologies.

 More particularly, this lysate can be obtained according to the technology described in US Pat. No. 4,464,362, and in particular according to the following protocol.

A probiotic microorganism considered is cultured anaerobically in a suitable culture medium, for example according to the conditions described in US 4,464,362. and EP 0 043 128. When the stationary phase of development is reached, the culture medium can be inactivated by pasteurization, for example at a temperature of 60 to 65 ° C for 30 min. The microorganisms may be collected by a conventional separation technique, for example membrane filtration, centrifuged and resuspended in a sterile solution of NaCl at a physiological concentration. In the context of a lysate, it can then be obtained by ultrasonic disintegration of such a medium in order to release the cytoplasmic fractions, the cell wall fragments and certain products derived from the metabolism of these microorganisms. Then all components in their natural distribution can be stabilized in a weakly acidic aqueous solution.

 It is thus possible to obtain a lysate having a concentration of about 0.1 to 50%, in particular from 1 to 20% and in particular about 5% by weight of active material (s) relative to its total weight. .

 According to a possible alternative to the technology described above, the centrifugation can be applied at the end of fermentation (at the beginning of the stationary phase) before the heat treatment.

 It is also possible to recover the microorganisms and the supernatant and subsequently carry out the heat treatment, the assembly can then be dried.

 The extract or lysate may be used in a variety of forms, such as a solution or in a powder form, preferably in the form of a solution. Storage in liquid form may require the application of a stabilizing treatment such as ultra-high temperature (UHT) treatment. Any method of stabilizing liquids known to those skilled in the art may be considered for the solution or suspension of Bifidobacterium longum.

 The lysate of Bifidobacterium longum used in the context of the invention may advantageously be the lysate registered under the name INCI: Bifidat ferment Lysate, under the name EINECS: Bifidobacterium longum, under the EINECS No.: No. 306-168-4 and under the CAS N °: 96507-89-0.

The product sold under the name Repair Complex CLR ^® by the company K. RICHTER GmbH and that is formed of an inactivated lysate of the species Bifidobacterium longum is particularly suitable for the implementation of the invention.

Probiotic microorganisms Bifidobacterium longum include subspecies Bifidobacterium longum subsp. infantis, Bifidobacterium longum subsp. longum, and Bifidobacterium longum subsp. am. Examples of suitable Bifidobacterium longum strains include Bifidobacterium longum subsp. longum BOR1, Bifidobacterium longum subsp. longum DJ010A, Bifidobacterium longum subsp. longum DM301, Bifidobacterium longum subsp. longum M58739, Bifidobacterium longum subsp. infantis 157F-NC, Bifidobacterium longum subsp. Infantis ATCC 15697, Bifidobacterium longum subsp. Infantis ATCC 55813, Bifidobacterium longum subsp. Infantis CCUG 52486, Bifidobacterium longum subsp. infantis JCM1217, Bifidobacterium longum subsp. infantis JCM1222, Bifidobacterium longum ATCC BAA-999, Bifidobacterium longum FERM BP-7787, Bifidobacterium longum ATCC 15707, Bifidobacterium longum ATCC 15708, Bifidobacterium longum ATCC 55817, Bifidobacterium longum FERM P-6548, Bifidobacterium longum CNCM 1-1228, strain Bifidobacterium longum filed under the Budapest Treaty to the National Collection of Cultures of Microorganisms (CNCM, Institut Pasteur, 28 rue du Dr Roux, 75724 Paris Cedex 15, France) under the number CNCM 1-2170, and the strain Bifidobacterium longum BB536.

 Preferably, the strain of Bifidobacterium longum used in the context of the invention is not the strain Bifidobacterium longum subsp. longum filed on 29 January 2001 with the CNCM (Paris, France) under the number 1-2618.

 Preferably, the strain of Bifidobacterium longum used in the context of the invention is the strain Bifidobacterium longum BB536.

 A probiotic microorganism Bifidobacterium longum used in the context of the invention may be formulated in a composition at a rate of at least 0.0001% expressed by dry weight, in particular at a rate of 0.0001 to 20% and more particularly at a rate of from 0.001 to 15%, in particular from 0.01 to 10%, and especially from 0.1 to 2% relative to the total dry weight of the composition containing it.

 A probiotic microorganism Bifidobacterium longum used in the context of the invention is in particular formulated in a composition intended to be administered orally.

In general, a composition according to the invention intended to be administered orally may comprise, for living Bifidobacterium longum microorganisms, from 10 ³ to 10 ¹⁵ cfu / g, in particular from 10 ⁵ to 10 ¹⁵ cfu / g and more. particularly from 10 ⁷ to 10 ¹² cfu / g live Bifidobacterium longum microorganisms per gram of support or equivalent doses calculated for inactivated or dead Bifidobacterium longum microorganisms or for Bifidobacterium longum microorganism fractions or for metabolites produced. In particular, in an orally administered composition, the concentration of microorganism and / or fraction and / or corresponding metabolite may be adjusted to correspond to doses, expressed in microorganism equivalent, ranging from 5x10 ⁵ to 10 ¹³ cfu / day (d) and in particular from 10 ⁸ to 10 ¹¹ cfu / d.

 It may be advantageous to use Bifidobacterium longum microorganisms in inactivated or even dead form, in particular in the form of a lysate or a cell extract, typically containing cell fragments and metabolites.

 A Bifidobacterium longum microorganism may also be included in a composition as fractions of cellular components or as metabolites, particularly as an extract or as a lysate. The microorganism (s) Bifidobacterium longum, metabolite (s) or fraction (s) can also be introduced in the form of a freeze-dried powder, a culture supernatant and / or or where appropriate in a concentrated form.

When a composition comprises metabolites, the contents of metabolites in the compositions correspond substantially to the contents that may be produced by the equivalent October ³ to October ¹⁵ cfu, particularly October ⁵ to October ¹⁵ cfu, more particularly ¹⁰ July at 10 ¹² cfu live Bifidobacterium longum microorganisms per gram of support.

 The expression of the amount of metabolites or fractions of an "ufc" microorganism, or of dead microorganisms, is intended to designate the quantity of this microorganism necessary for the production of said quantity of metabolites or fractions of microorganisms.

Cosmetic composition

 In a particular embodiment of the invention, said at least one strain of Bifidobacterium longum, as defined in the section "Bifidobacterium longum" above, is formulated in a cosmetic composition suitable for oral administration, comprising in particular besides an unmanageable vehicle.

 By "unmanageable vehicle" is meant here a medium suitable for oral administration which does not cause undesirable effects during or following its administration, in particular during or following ingestion by an individual. .

 The ingestible vehicle will be adapted to the form in which the composition is intended to be packaged, especially solid or fluid at room temperature and atmospheric pressure.

The cosmetic composition used in the context of the invention may be in any of the galenical forms normally used for the oral route of administration. A cosmetic composition used in the context of the invention may constitute a composition for treating or caring for the skin or the scalp, keratinous fibers such as the hair, the eyelashes or the eyebrows, or a sun protection or tanning composition. artificial. Preferably, the cosmetic composition used in the context of the invention is a sun protection or artificial tanning composition.

 Such a composition may optionally contain additional cosmetic active agents, especially as indicated below.

 In the case of a cosmetic composition for oral administration, and in particular comprising a probiotic microorganism Bifidobacterium longum as defined in the section "Bifidobacterium longum" above, the use of an ingestible support is preferred.

 The unmanageable support may be of various nature depending on the type of composition considered.

 Particularly suitable as food carriers are tablets, capsules or lozenges, oral supplements in dry form and oral supplements in liquid form.

 It may for example be food supplements, the formulation of which may be carried out by the usual methods for producing in particular an oral solution, dragees, capsules, gels, emulsions, swallowing or chewable tablets, capsules , including soft or hard capsules, granules for dissolving, syrups, solid or liquid foods and hydrogels for controlled release.

 According to one embodiment, a cosmetic composition, implemented in the context of the invention and administered orally, may be formulated in the form of dragees, capsules, gels, emulsions, tablets, capsules, of hydrogels, food bars, powders, compacted or not, suspensions or liquid solutions, confectionery, fermented milks, fermented cheeses, chewing gum, toothpaste or spray solutions.

When the composition is an emulsion, the proportion of the fatty phase can range from 5 to 80% by weight, and preferably from 5 to 50% by weight relative to the total weight of the composition. The oils, emulsifiers and coemulsifiers used in the composition in emulsion form are chosen from those conventionally used in the cosmetics and / or dermatological field. The emulsifier and the coemulsifier may be present in the composition in a proportion ranging from 0.3 to 30% by weight, and preferably from 0.5 to 20% by weight relative to the total weight of the composition. When the oral composition used in the context of the invention is an oily solution or gel, the fatty phase may represent more than 80% of the total weight of the composition.

 In particular, the probiotic microorganism Bifidobacterium longum used in the context of the invention can be incorporated into any form of food supplements or fortified foods, for example food bars or powders, compacted or not. The powders can be diluted in water, soda, dairy products or soy derivatives, or incorporated into food bars.

 A probiotic Bifidobacterium longum microorganism of the invention, as defined in the "Bifidobacterium longum" section above, may be furthermore formulated with the usual excipients and components for such oral compositions or food supplements, namely in particular fatty components. and / or aqueous, humectants, thickeners, preservatives, texture, flavor and / or coating agents, antioxidants, preservatives and dyes customary in the field of food.

 The formulating agents and excipients for oral composition, and especially for food supplements, are known in the art and are not the subject of a detailed description here.

 Milk, yoghurt, cheese, fermented milk, fermented milk products, ice cream, cereal-based products or fermented cereal products, powders containing milk, formulas for children and infants, food products of the confectionery type, chocolate, cereals, especially pet food, tablets, capsules or tablets, suspensions of liquid bacteria, oral supplements in dry form and supplements oral in liquid form.

 In a particularly preferred embodiment of the invention, the cosmetic composition as defined above further comprises an additional cosmetic active.

 Advantageously, such additional asset may be intended to exert a cosmetic or caring effect on the skin, the hair, the eyelashes, the hairs and / or the scalp.

 The additional active agents are chosen by those skilled in the art so that they do not interfere with the effect of the probiotic Bifidobacterium longum microorganisms of the invention.

According to one variant embodiment, a probiotic Bifidobacterium longum microorganism, as defined above in the "Bifidobacterium longum" section, can advantageously be used with, as additional active ingredient, a non-probiotic microorganism, in particular the strain Vitreoscilla filiformis. Such a non-probiotic microorganism can be implemented in living, semi-active, inactivated or dead form, in particular as defined in the "Bifidobacterium longum" section above. In particular, a microorganism of the Vitreoscilla filiformis type, as described in particular in the patent application FR 0 625 782, may be used in a living form or in a fractionated or complete lysate form with its culture medium after evaporation. concentration of the H ₂ 0 composition.

In known manner, the dosage forms dedicated to oral administration may also contain adjuvants which are customary in the cosmetic, pharmaceutical and / or dermatological field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, perfumes, fillers, filters, bactericides and dyes. The amounts of these various adjuvants are those conventionally used in the field under consideration, and for example from 0.01 to 20% of the total weight of the composition. These adjuvants, depending on their nature, can be introduced into the fatty phase and / or into the aqueous phase.

 Fats which can be used in the invention include mineral oils such as, for example, hydrogenated polyisobutene and liquid petroleum jelly, vegetable oils such as, for example, a liquid fraction of shea butter, sunflower oil and almonds. apricot, animal oils such as perhydrosqualene, synthetic oils including Purcellin oil, isopropyl myristate and ethyl hexyl palmitate, unsaturated fatty acids and fluorinated oils such as perfluoropolyethers. It is also possible to use fatty alcohols, fatty acids such as stearic acid and, for example, waxes such as paraffin, carnauba and beeswax. It is also possible to use silicone compounds such as silicone oils and, for example, cyclomethicone and dimethicone, waxes, resins and silicone gums.

As emulsifiers that can be used in the invention, mention may be made, for example, of glycerol stearate, polysorbate 60, cetylstearyl alcohol / cetylstearyl alcohol mixture oxyethylenated with 33 moles of ethylene oxide sold under the name Sinnowax AO® by Henkel. the mixture of PEG-6 / PEG-32 / glycol stearate sold under the name Tefose® 63 by Gattefosse, PPG-3 myristyl ether, silicone emulsifiers such as cetyldimethicone copolyol and sorbitan mono- or tristearate, PEG-40 stearate, oxyethylenated sorbitan monostearate (20OE). As solvents that can be used in the invention, mention may be made of lower alcohols, in particular ethanol and isopropanol, and propylene glycol.

 The composition of the invention may also advantageously contain a thermal and / or mineral water, in particular chosen from Vittel water, the waters of the Vichy basin and Roche Posay water.

 As hydrophilic gelling agents, mention may be made of carboxylic polymers such as carbomer, acrylic copolymers such as acrylate / alkylacrylate copolymers, polyacrylamides and in particular the polyacrylamide, C13-14-lsoparaffin and Laureth-7 mixture sold under the name Sepigel 305® by the company SEPPIC, polysaccharides such as cellulose derivatives such as hydroxyalkylcelluloses and in particular hydroxypropylcellulose and hydroxyethylcellulose, natural gums such as guar, carob and xanthan and clays.

 As lipophilic gelling agents, mention may be made of modified clays such as bentones, metal salts of fatty acids such as aluminum stearates and hydrophobic silica, or else ethylcellulose and polyethylene.

Of course, the topical compositions according to the invention may furthermore contain several other active ingredients.

 As assets conventionally used, mention may be made of vitamins B3, B5, B6, B8, C, E, or PP, niacin, carotenoids, polyphenols and minerals such as zinc, calcium and magnesium. ..

 In particular, it is possible to use an antioxidant complex comprising vitamins C and E, and at least one carotenoid, in particular a carotenoid chosen from β-carotene, lycopene, astaxanthin, zeaxanthin and lutein, flavonoids such as as catechins, hesperidin, proanthocyanidins and anthocyanins.

 It can also be at least one prebiotic or a mixture of prebiotics. More particularly, these prebiotics can be chosen from oligosaccharides, produced from glucose, galactose, xylose, maltose, sucrose, lactose, starch, xylan, hemicellulose, inulin, acacia-type gums, for example, or of their mixtures. More particularly, the oligosaccharide comprises at least one fructooligosaccharide. More particularly, this prebiotic may comprise a mixture of fructo-oligosaccharide and inulin.

With regard to lipophilic active agents, it is possible to use retinol (vitamin A) and its derivatives, tocopherol (vitamin E) and its derivatives, ceramides, essential oils and unsaponifiables (tocotrienol, sesamin, gamma oryzanol, phytosterols, squalens, waxes, terpenes). Other active agents that may be more particularly associated with the lysate in an oral dosage form may also be all the commonly used and / or authorized ingredients.

 By way of illustration, mention may be made of vitamins, minerals, essential lipids, trace elements, polyphenols, flavonoids, phytoestrogens, antioxidants such as lipoic acid and coenzyme Q10, carotenoids, probiotics, prebiotics , proteins and amino acids, mono- and polysaccharides, amino-sugars, phytosterols and triterpene alcohols of vegetable origin.

 These are, in particular, vitamins A, C, D, E, PP and group B. Of the carotenoids, beta-carotene, lycopene, lutein, zeazanthin and astaxanthin are preferably selected. . The minerals and trace elements particularly used are zinc, calcium, magnesium, copper, iron, iodine, manganese, selenium, chromium (III). Among the polyphenols, polyphenols of grape, tea, olive, cocoa, coffee, apple, blueberry, elderberry, strawberry, cranberry, and onion are also particularly preferred. Preferably, among the phytoestrogens, isoflavones are retained in the free or glycosylated form, such as genistein, daidzein, glycitein or lignans, in particular those of flax and schizandra chinensis. Probiotics are preferably selected from the group consisting of lactobacilli and bifidobacteria. Amino acids or peptides and proteins containing them, such as taurine, threonine, cysteine, tryptophan, methionine. The lipids preferably belong to the group of oils containing monounsaturated and polyunsaturated fatty acids such as oleic, linoleic, alpha-linolenic, gamma-linolenic, stearidonic, long-chain omega-3 fatty acids such as ΙΈΡΑ and the like. DHA, conjugated fatty acids derived from plants or animals such as CLA (Conjugated Linoleic Acid).

Non-therapeutic cosmetic use

 Surprisingly, the inventors have observed that the administration of probiotic microorganisms of the Bifidobacterium longum type regulates melanocyte homeostasis by promoting the synthesis of melanin, more effectively than other probiotic microorganisms such as Propionibacterium freudenreichii or Bacillus coagulans. . Thus the natural pigmentation of the skin, hair and hair is strengthened. The cutaneous or capillary pigmentation thus obtained is advantageously homogeneous.

The present invention relates more particularly to the non-therapeutic cosmetic use of a strain of Bifidobacterium longum, as defined in section "Bifidobacterium longum" above, optionally formulated in a cosmetic composition as defined in the section "Cosmetic composition" above, as an agent inducing cutaneous and / or capillary pigmentation, for physiologically inducing pigmentation and thus in particular preventing and and / or treating an alteration of the homogeneity of the color of the skin and / or the hair, said at least one strain of Bifidobacterium longum being administered orally.

 The cosmetic composition used in the context of the invention can in particular be used in healthy subjects, typically subjects that do not have any alterations in the homogeneity of the skin color and / or the hair of origin. pathological.

 It can, especially in healthy subjects, be used as a tanning agent, advantageously as a tanning agent independent of sunlight and / or UV rays, or to accelerate and / or intensify the tan.

 It can also, especially in these healthy subjects, be used to make the complexion and / or the color of the complexion more homogeneous.

 The cosmetic composition used in the context of the invention may also be used to prevent and / or cosmetically treat a cutaneous and / or capillary pigmentation defect resulting from an imbalance in melanocyte homeostasis.

 The imbalance of melanocyte homeostasis may be an imbalance in proliferation and / or maturation and / or survival of melanocytes.

 Proliferation, maturation and survival concern the different life stages of a cell in a tissue of an organism, ie its division from a stem cell or a mother cell, its differentiation allowing it to acquisition of characters specific to the tissue in which it fits, such as the expression of the different enzymes and proteins necessary for melanogenesis, and its maintenance until its senescence and apoptosis. Dysregulation of these steps may result in an imbalance in the distribution of melanocytes in the epidermis or in the follicle, or an imbalance in melanogenesis affecting, for example, the natural color of the epidermis, hair or hair.

 In a particular embodiment, the pigmentation defects concerned by the invention result from an imbalance of melanogenesis.

The imbalance of melanocyte homeostasis resulting in the pigmentation defects concerned by the invention may also be associated with skin aging or scalp, and in particular be aggravated by this aging. The pigmentation disorders concerned by the invention may relate to the epidermis, the scalp, the hair, the hairs or the eyelashes.

 By way of cosmetic hair pigmentation defects referred to by the invention, mention may in particular be made of canities.

 Canitia is a natural hair whitening that occurs with age, and is primarily associated with a decrease in melanin in the hair shaft. More specifically, canities are linked to a specific and progressive rarefaction of hair melanocytes, affecting both the hair bulb melanocytes and the melanocyte precursor cells. Advantageously, the implementation of the invention makes it possible to reduce or even to slow down or delay the appearance of the white hair and the graying of the hair. Advantageously, the present invention makes it possible to stimulate, or even reactivate, melanogenesis, in order to delay, reduce or even reduce canities.

 The composition used in the context of the invention may also be used in subjects having alterations in the homogeneity of the color of the skin and / or the hair, in particular subjects with skin pigmentation defects and / or or cosmetic capillary related to an accumulation, in particular heterogeneous melanin, to homogenize the color of the skin and / or hair.

 Such cutaneous and / or capillary pigmentation defects related to a heterogeneous accumulation of melanin include melasma, acne pigment sequelae, post-inflammatory pigmentation, depigmentations due to burns or surgery and benign facial dyschromias.

 In these applications, the compositions used in the context of the invention make it possible to reduce the observed dyschromias and to homogenize the color of the skin, in particular by increasing or improving the pigmentation of the skin around the hyperpigmented areas and / or by increasing or improving the pigmentation of the skin in hypopigmented areas.

 Preferably, the alteration of the homogeneity of the skin and / or hair targeted by the present invention is selected from the group consisting of canities, melasma, acne pigment sequelae, post-inflammatory pigmentation, depigmentation due to burns or surgery and benign facial dyschromias.

The present invention also relates to a non-therapeutic cosmetic treatment method for physiologically inducing the pigmentation of the skin and / or the hair, and thus to homogenize the color of the skin and / or the hair, and in particular to prevent and / or treat a alteration of the homogeneity of the skin color and / or hair, in which a cosmetically effective amount of a strain of Bifidobacterium longum, as defined in the section "Bifidobacterium longum" above, optionally formulated in a cosmetic composition as defined in the section, is administered orally to a subject "Cosmetic composition" above.

 By "cosmetically effective amount" is meant here a sufficient amount of agents used in the context of the invention to treat and / or prevent said cosmetic disorder, which does not generate unacceptable side effects for the user.

 This strain of Bifidobacterium longum, optionally formulated in a cosmetic composition, is administered orally.

 Preferably, the administration of the cosmetic composition according to the invention is in the form of a dietary supplement.

Therapeutic applications

 The present invention also relates to a method for preventing and / or treating an alteration of the homogeneity of the color of the skin and / or the hair of pathological origin in a subject, comprising the administration, orally, in a subject in need of a therapeutically effective amount of a probiotic microorganism strain Bifidobacterium longum as defined in the section "Bifidobacterium longum" above.

 The present invention also relates to the use of a strain of Bifidobacterium longum as defined in the "Bifidobacterium longum" section above, for the manufacture of an orally administrable pharmaceutical composition intended for the prevention and / or treatment of an alteration of the homogeneity of the color of the skin and / or hair of pathological origin.

 As used in the context of the embodiments of the invention associated with a therapeutic use of the probiotic microorganisms Bifidobacterium longum defined in the section "Bifidobacterium longum" above, the term "alteration of the homogeneity of the color of the skin and / or hair of pathological origin "refers to an alteration of the homogeneity of the skin and / or hair color observed in dermatological conditions resulting from a deficiency or absence of melanin or a heterogeneous distribution of melanin, in particular dyspigmentation.

By "dyspigmentation" is meant an abnormality of pathological origin in the formation or distribution of melanin at the cutaneous or capillary level. The cutaneous and / or capillary diseases of pigmentation deviate from the cosmetic field by the degree of severity and the intensity of the symptoms by which they manifest themselves.

 In the case of alterations resulting from a deficit or absence of melanin, the probiotic microorganisms Bifidobacterium longum defined above will make it possible to homogenize the color of the skin, in particular by increasing and / or improving the pigmentation of the skin. skin in depigmented or hypopigmented areas.

 Such alterations include in particular the alterations in the homogeneity of the skin color associated with vitiligo.

 In the case of alterations resulting from an excess of melanin, the probiotic microorganisms Bifidobacterium longum defined above will make it possible to homogenize the color of the skin, in particular by increasing and / or improving the pigmentation of the skin around the hyperpigmented zones. .

 Such alterations include, in particular, alterations in the homogeneity of the skin color associated with actinic lentigines or with the dermatitis of the meadows.

 According to one embodiment, the alterations in the homogeneity of the color of the skin and / or hair of pathological origin are chosen from actinic lentigo, meadow dermatitis and vitiligo. In the following description and examples, unless otherwise indicated, percentages are percentages by weight and ranges of values in the form "between ... and ..." include the specified lower and upper bounds.

 The ingredients are mixed before being shaped in the order and under conditions easily determined by those skilled in the art.

 The content and the nature of the ingredients used in the compositions of the invention are adjusted by those skilled in the art so as not to substantially affect the properties required for the compositions of the invention.

The following examples are presented by way of illustration and not limitation of the field of the invention. Brief description of the figure

The Figure is a set of histograms showing the melanin level, in percent relative to the control, quantified in co-cultures of primary keratinocytes (NHEK) and primary melanocytes (NHEM) incubated for 72 h with culture medium alone ( control), from ΙΊΒΜΧ to 200 μΜ (IBMX) as a positive control, a lysate of Bifidobacterium longum (B. longum CLR 1%), a lysate of Propionibacterium freudenreichii (P. freundenreichii 0.2%) and a lysate of Bacillus coagulans ( B. coagulants 0.2 mg / ml).

Examples

Example 1 Induction of melanin synthesis by Bifidobacterium longum

This example demonstrates the pro-pigmenting activity of the probiotic microorganism Bifidobacterium longum.

Material and methods

Primary human melanocytes (NHEM) were inoculated into 24-well plates in the presence (co-culture) of normal human epidermal keratinocytes (NHEK) and then cultured for 24 h. The cells were then treated with nothing (negative control), IBMX (positive control, 200 μΜ), an extract of Bifidobacterium longum (Repair Complex CLR ^® 1%), an extract from Propionibacterium freudenreichii (0.2%) or an extract Bacillus coagulans (2 mg / ml).

 After treatment, the cells were incubated for 72 h, all experimental conditions having been performed in triplicate. After 72 hours of incubation, the cells were lysed with a 0.5 N NaOH solution in order to extract the melanin. The optical density of the samples was measured at 405 nm, with reference to an exogenous melanin range (standard melanin curve 0.39 to 100 μg / ml).

 The results are expressed as a percentage of relative stimulation compared with the control grown in standard co-culture medium. They were analyzed with Student's statistical test:

^* : 0.01 <p <0.05 significant;

* ^* : 0.001 <p <0.01 very significant;

^*** : p <0.001 extremely significant Results

 The inventors have thus shown that the incubation of NHEK / NHEM co-cultures with lysates of probiotic microorganisms made it possible to significantly increase melanin production by melanocytes compared with untreated cocultures (see FIG. .

 They have also, surprisingly, shown that the specific use of lysates of strains of Bifidobacterium longum makes it possible to increase even more significantly the production of melanin compared to the use of lysates of other probiotics such as P freudenreichii or B. coagulans. Indeed, incubation of NHEK / NHEM co-cultures with B. longum CLR lysates induces an increase in melanin production of about 1, 4-fold compared to untreated control, whereas lysates of P. freudenreichii and B. coagulans only achieve an increase of about 1, 1 time. It should be noted that the increase observed with B. longum is even greater than that obtained with the IBMX positive control.

 Thus these results demonstrate that the probiotic microorganism B. longum can be used very effectively as a pro-pigmenting agent.

Example 2 Compositions according to the invention for the oral route

Example 2A: Stick powder

Bifidobacterium longum 10 ⁰ ufc

 Calcium citrate 50 mg

 Xanthan gum 0.8 mg

 Sodium benzoate 0.2 mg

 Maltodextrin qs 30 g

 You can take a stick a day.

Example 2B: Capsule

Bifidobacterium longum 10 ⁹ ufc

Magnesium gluconate 150 mg / capsule

Vitamin C 60 mg / capsule

 Magnesium stearate 0.02 mg / capsule

 One to three of these capsules can be taken daily.

Claims

1. Non-therapeutic cosmetic use of at least one strain of probiotic microorganism Bifidobacterium longum as agent inducing cutaneous and / or capillary pigmentation, for homogenizing the color of the skin and / or the hair, said at least one strain of probiotic microorganism Bifidobacterium longum being administered orally and said at least one strain of Bifidobacterium longum not being the strain Bifidobacterium longum subsp. longum filed on 29 January 2001 with the CNCM (Paris, France), under the number 1-2618. 2. Cosmetic use according to claim 1, wherein said strain of Bifidobacterium longum is in the form of a lysate. 3. The cosmetic use according to claim 2, wherein said lysate is lysate sold under the name Repair Complex CLR ^® by the company K. RICHTER GmbH. 4. Cosmetic use according to any one of claims 1 to 3, for preventing and / or treating an alteration of the homogeneity of the color of the skin and / or hair. 5. Cosmetic use according to claim 4, wherein the alteration of the homogeneity of the color of the skin and / or hair is selected from the group consisting of canities, melasma, acne pigment sequelae, post-inflammatory pigmentation, depigmentation due to burns or surgery and benign facial dyschromias. 6. Cosmetic use according to any one of claims 1 to 5, wherein said strain of Bifidobacterium longum is formulated in a cosmetic composition further comprising an ingestible vehicle. The cosmetic use of claim 6, wherein the cosmetic composition further comprises an additional cosmetic active. 8. Bifidobacterium longum probiotic microorganism strain for its use as a skin and / or capillary pigmentation inducing agent, for the prevention and / or treatment of an alteration of the homogeneity of the color of the skin and / or hair of pathological origin, said strain of probiotic microorganism Bifidobacterium longum being administered orally and said strain of Bifidobacterium longum n ' being not the strain Bifidobacterium longum subsp. longum filed on 29 January 2001 with the CNCM (Paris, France) under the number 1-2618. 9. Strain of Bifidobacterium longum for its use according to claim 8, said alteration of the homogeneity of the color of the skin and / or the hair of pathological origin being selected from the group consisting of actinic lentigo, of the dermatitis of the meadows. and vitiligo. 10. Bifidobacterium longum strain for use according to claim 8 or 9, said strain of Bifidobacterium longum being in the form of a lysate. 11. Bifidobacterium longum strain for use according to claim 10 wherein the lysate is a lysate sold under the name Repair Complex CLR ^® by the company K. RICHTER GmbH.