FR3090384A1 - Pro-vitamin effect of an extract of a probiotic microorganism - Google Patents

Abstract

The present invention relates to the use of at least one extract of a probiotic microorganism, one of its fractions and / or one of its metabolites, in a composition for topical application to keratin materials, as a provitamin agent.

Description

Description

Title of the invention: Pro-vitamin effect of an extract of a probiotic microorganism

Technical area

The present invention relates to the use of an extract of a probiotic microorganism in a composition for topical application, as an agent with provitamin effect.

Prior art

It is well known that the quality of nutrition is reflected in the health of the skin and that certain skin disorders are associated with deficiencies, in particular in certain micronutrients (Park K. Role of micronutrients in skin health and function. Biomol .Ther. 2015; 23; 207-217).

The micro-nutritional balance of the skin can indeed be affected by external and / or internal factors, such as, for example, fatigue, stress, oxidizing effects, UV exposure, and / or senescence cells.

Micronutrients are essential for the development and formation of the skin (organ in constant evolution), the physiological renewal of the epidermis and the adaptation of the skin to its environment (role of protective envelope). Vitamins in particular participate in particular:

- antioxidant protection (vitamins A, E, C)

- lipid, carbohydrate or protein metabolism (vitamins B3, B5, C for collagen synthesis), energy production (vitamin B2),

- normal blood clotting (vitamin K) and calcium absorption for good bone health (vitamin K).

The use of vitamins (A, E, C) is known in the formulation of cosmetic compositions intended for the skin, in particular for their protective and / or nutritional effect, but there remains the need for new compounds making it possible to maintain and / or promote the nutritional balance of the skin.

Unexpectedly, the Applicant has demonstrated on skin cells in vitro that an extract of a probiotic microorganism, in particular of the species Lactobacillus rhamnosus (CNCM I-5313), has an action on the vitamin transport and vitamin receptors (activation of 12 genes) of group B vitamins, vitamin A (retinol), vitamins K and vitamins C and E. This extract from a probiotic microorganism also has a stimulating effect on transport of energy sugars (GLUT-1 = SLC2A1, GLUT-3 = SLC2A3).

The use of such an extract in a composition for topical application to keratin materials therefore makes it possible to impart a nutritional and pro-vitamin effect to keratin materials, in particular the skin, resulting from the stimulation of several carriers and receptors vitamins in the skin cells and which are directly involved in the beneficial effects of these vitamins for the skin, and an anti-fatigue and toning effect (energy production, transport of energy sugars and pro-vitamin action).

It can also be advantageous to combine the extract of a microorganism according to the invention with other vitamins or extracts containing it, administered as a topical or oral supplement (food supplement).

Statement of the invention

A first object of the invention is therefore the use of at least one extract of a probiotic microorganism, of one of its fractions and / or of one of its metabolites, in a composition for application topically on keratin materials, as a provitamin agent, capable in particular of promoting and / or improving the nutrition of keratin materials and / or vitamin efficiency.

According to a preferred embodiment, the probiotic microorganism extract used in the invention is a Lactobacillus rhamnosus lysate, in particular a Lactobacillus rhamnosus lysate (CNCM 1-5313).

The present invention also relates to a non-therapeutic cosmetic process intended to promote and / or improve the nutrition of keratin materials, comprising the application to keratin materials, in particular the skin and / or its integuments and / or the lips , at least one extract of a probiotic microorganism according to the invention, preferably a Lactobacillus rhamnosus lysate (CNCM 1-5313), or a cosmetic composition for topical application containing it.

In particular, the method is intended in particular for tired, aged, dull skin, impacted by an unfavorable or unbalanced nutritional context, emotional stress or lack of sleep.

Another object of the invention is a composition for topical application to keratin materials comprising, in a physiologically acceptable medium, at least one extract of a probiotic microorganism as defined in the invention, preferably a lysate of Lactobacillus rhamnosus (CNCM 1-5313), and at least one vitamin or extract containing it, in particular vitamin A, C, E and their mixtures.

Another object of the invention is a cosmetic assembly comprising at least a first composition for topical application comprising an extract of a probiotic microorganism as defined in the invention, preferably a lactate of Lactobacillus rhamnosus (CNCM 1- 5313), and at least a second composition for topical application or oral administration, preferably a food supplement, comprising a vitamin or a plant extract containing it, in particular a vitamin A, C, E and their mixtures.

Description of the embodiments

The invention therefore relates in particular to the use of at least one extract of a probiotic microorganism, one of its fractions and / or one of its metabolites, in a composition for topical application to keratin materials, as a provitamin agent, capable in particular of promoting and / or improving the nutrition of keratin materials and / or vitamin efficiency.

By "keratin materials" is meant according to the invention the skin, the mucous membranes (in particular the lips), and the integuments. According to a particular embodiment, the composition is intended for topical application to the skin, the lips or the integuments, preferably the skin.

By "physiologically acceptable medium" is meant a medium compatible with keratin materials, in particular the skin and / or the lips.

By 'pro vitamin agent' or 'pro vitamin effect' according to the invention, it is meant that the extract of a probiotic microorganism according to the invention has an effect on (i) the activation of vitamin transporters (ex : ABCA1, SLC19A3, SLC2A3 and SLC52A1) and / or on (ii) activation of vitamin receptors.

The activation of vitamin transporters allows better bioavailability in the skin of the vitamins concerned and their use by the skin cells and the activation of vitamin receptors facilitates the biological action of these vitamins on the skin cells.

Among the vitamins concerned by the stimulating action (at the level of the genes) of the extract of an L rhamnosus microorganism according to the invention, on their transporters and receptors, the Applicant has identified:

- Vitamin B1 (SLC19A3)

- Vitamin B2 (SLC52A1),

- Vitamin B12 (CUBN, TCN1 and 2)

- Vitamin B9 (FOLR3),

- Vitamin B3 (HCAR2 and 3),

- Vitamin A / retinol (RBP1, STRA6),

- vitamins E and K (SCARB1),

- vitamins K (ABCA1),

- Vitamin C (SLC2A3)

Thus, the extract of a probiotic microorganism according to the invention, preferably the Lactobacillus rhamnosus lysate (CNCM 1-5313) is capable in particular of promoting and / or improving the nutrition of keratin materials and the vitamin efficiency . Probiotic microorganism extract

According to the definitions used by international scientific associations for probiotics and prebiotics, the term "probiotics" means living microorganisms which, when administered in adequate quantity, have beneficial effects on the health of host. The term "prebiotics" means food ingredients resistant to digestion which induce specific changes in the composition and / or activity of the intestinal microbiota, thus producing a beneficial effect on the health of the host. By "symbiotics" is meant products that contain both probiotics and prebiotics.

Or the microorganism (s) little (Fri) t be included in a composition according to the invention in a living form, semi-active or inactivated, dead. They can be included as fractions of cellular components or as metabolites. The microorganism (s), metabolite (s) or fraction (s) can also be introduced in the form of a lyophilized powder, a culture supernatant and / or the if necessary in a concentrated form.

According to a particular embodiment, these microorganisms are used in an inactivated or even dead form, and more particularly in the form of a lysate.

Thus, according to a particular and preferred mode, the extract of a probiotic microorganism according to the invention is a lysate of probiotic microorganism (not living), one of its fractions and / or one of its metabolites.

We will talk indifferently in the description 'probiotic extract' or 'probiotic microorganism extract' or 'probiotic bacterial strain extract' or 'probiotic lysate' or 'microorganism lysate probiotic 'or' probiotic bacterial strain lysate '.

By "lysate" is meant in particular a material obtained by cell lysis of the cells of the microorganism under consideration thus causing the release of the intracellular biological constituents naturally contained in said cells. The lysate used is formed in whole or in part of the intracellular biological constituents and of the constituents of the cell walls and membranes. The present invention therefore relates to the use of a probiotic lysate and / or one of its fractions.

Within the meaning of the invention, the term "fraction" more particularly designates a fragment of said microorganism endowed with provitaminic efficacy by analogy to said whole microorganism.

In particular, the lysate according to the invention contains the cellular cytoplasmic fraction containing the enzymes (ex: dehydrogenases, phosphatases ...) and / or the constituents of the cell walls (ex: peptidoglycans ...) and / or the constituents of cell membranes (e.g. glycerophospholipids ...).

Cell lysis can be carried out by various technologies well known to those skilled in the art, such as for example osmotic shock, thermal shock, by ultrasound, or even under mechanical stress (eg centrifugation).

According to a preferred embodiment, the lysate is obtained by centrifugation and osmotic shock (ex: addition of sodium hydroxide).

According to a particular mode, the microorganism is cultured anaerobically in an adequate culture medium. 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 are then collected by a conventional separation technique, for example membrane filtration, centrifugation and resuspended in a sterile physiological NaCl solution. The lysate can be obtained by ultrasonic disintegration of such a medium in order to release the cytoplasmic fractions, the cell wall fragments and the products derived from metabolism. Then all the components in their natural distribution are then stabilized in a weakly acidic aqueous solution.

The lysate thus obtained generally has a concentration of the order of 0.1 to 10%, in particular from 0.5 to 5% and in particular from 1 to 2% by weight in dry matter (s) (active) compared to its total weight.

The lysate can be used in different forms, in the form of a solution or in the form of a powder.

More particularly, the probiotic microorganism comes from the group of lactic acid bacteria, such as in particular Lactobacillus and / or Bifidobacterium. By way of illustration of these lactic bacteria, mention may be made more particularly of Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei or Bifidobacterium bifidum Bifidobidium Bifidifidumidumid Bacteridobidium Bifidobidium Bifidobidium Bifidobidium Bifidobidium Bifidobidium pseudocatenulatum, and mixtures thereof.

In a particular mode, the probiotic microorganism belongs to the genus Lactobacillus, to the genus Bifidobacterium, or their mixture, preferably to the genus Lactobacillus chosen from the group consisting of: Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus casei, Lactobacillus rhamnosus, , and Lactobacillus johnsonii.

The most suitable species are Lactobacillus johnsonii, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei, Bifidobacterium adolescentis, Bifidobacterium longum and respectively deposited according to the Budapest Treaty with the Pasteur Institute (28 rue du Docteur Roux, F-750 Paris cedex 15) on 06/30/92, 01/12/99, 04/15/99 and 04/15/99 under the following designations CNCM I6

1225 (Lactobacillus johnsonii), CNCM 1-2116 (Lactobacillus paracasei), CNCM I2168 (Bifidobacterium) and CNCM 1-2170 (Bifidobacterium longum) and the genus Bifidobacterium lactis (Bb 12) (ATCC27536) on Bifidobacterium longum (BB5) information described in application FR2938429.

According to a particular mode, the lysate of Bifidobacterium longum perhaps that registered under the name INCI: Bifidat ferment Lysate, under the name EINECS: Bifidobacterium longum, under the N ° EINECS: N ° 306-168-4 and under CAS number: No. 96507-89-0.

According to another particular and preferred embodiment, a microorganism of the genus Lactobacillus sp. Is used, in particular chosen from the species Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus casei, and their mixtures.

According to a particular embodiment, one can use a Lactobacillus paracasei, such as the strain Lactobacillus paracasei ST11 deposited according to the Budapest Treaty with the Institut Pasteur (28 rue du Docteur Roux, F-75024 Paris cedex 15) 01/12/99 under the designation CNCM 1-2116.

According to another particular and preferred embodiment, the probiotic microorganism according to the invention belongs to the species Lactobacillus rhamnosus (ATCC 7469, CCM 1825, NCDO 243, NCIB 6375 or even WDCM 00101 according to the collections in which it has been deposited), preferably the strain Lactobacillus rhamnosus CNCM 1-5313, deposited according to the Budapest Treaty with the Institut Pasteur (28 rue du Docteur Roux, F-75024 Paris cedex 15) on March 8, 2018 under the designation CNCM 1 -5313, and / or one of its fractions and / or one of its metabolites.

A lysate of this L. rhamnosus strain is obtained according to the following protocol from the supplier Greentech:

1. Culture of the L. rhamnosus strain in a suitable culture medium;

2. Biomass recovery by centrifugation;

3. Cell lysis (addition of soda);

4. Adjustment of the pH (addition of citric acid) and of the dry matter (addition of sterile water) to obtain an active ingredient content ranging from 0.1% to 10% by weight of dry matter (active ingredient) per relative to the total weight of said lysate;

5. Addition of preservative (phenoxyethanol).

Such a L. rhamnosus lysate is called Probiophyte LB by the supplier Greentech, and defined in INCI name under the name WATER and LACTOBACILLUS FERMENT LYSATE and PHENOXYETHANOL and SODIUM HYDROXIDE and CITRIC ACID, phenoxyethanol can be replaced by another preservative .

This lysate comprises a content of Lactobacillus rhamnosus ranging from 0.1% to 10%, in particular from 0.5 to 5% by weight and in particular from 1 to 2% by weight of dry matter (active material) relative the total weight of said lysate.

The microorganism lysate of the invention can be formulated in a composition in an amount of at least 0.0001% (expressed in dry weight or weight of active material), in particular in an amount of 0.0001 to 10% and more particularly at a rate of 0.0001 to 5% by weight, in particular from 0.0001 to 2% by weight, and in particular from 0.0001% to 1%, or even from 0.0001 to 0.05% by weight of active material (dry matter) relative to the total weight of the composition.

The probiotic microorganism lysate according to the invention can be used alone or in combination with other probiotic microorganisms, prebiotic ingredients, symbiotic ingredients or their mixture.

According to another particular mode, the lysate of probiotic microorganism according to the invention can be used in combination with vitamins or extracts containing it, said vitamins possibly being present in the composition of the invention or a separate composition, for a topical or oral application (food supplement), administered simultaneously, sequentially or delayed over time, as described below.

With this vitamin intake, it is also possible to associate other cosmetic active ingredients chosen from oils rich in essential fatty acids or not, separate extracts from the probiotic extract according to the invention and capable of stimulating vitamin receptors and transporters. , prebiotic extracts (which facilitate the growth of certain beneficial bacterial species of the cutaneous microflora at the expense of undesirable species), other extracts of other strains of probiotic microorganisms or even symbiotic ingredients (probiotic extract + prebiotic extracts) , seboregulators (to favor the biodiversity of the flora) and firming agents (to limit the enlargement of pores), soothing agents, protective active agents in particular stimulators of antioxidant and detoxifying defenses of the skin, depigmenting agents, desquamating agents, antiaging active ingredients, moisturizing ingredients, immunomodulating active agents readers, and mixtures thereof.

Cosmetic process

The invention also relates to a non-therapeutic cosmetic process intended to promote and / or improve the nutrition of keratin materials, comprising the application to keratin materials, in particular the skin and / or its integuments and / or the lips , at least one extract of a probiotic microorganism according to the invention or of a cosmetic composition for topical application containing it.

According to a preferred embodiment, the extract of a probiotic microorganism according to the invention is a Lactobacillus rhamnosus lysate, in particular a Lactobacillus rhamnosus lysate (CNCM 1-5313).

According to a particular embodiment of the invention, one can associate with the probiotic microorganism extract according to the invention, in the same composition or in two separate compositions intended to be used simultaneously, sequentially or delayed over time , topically or orally (eg food supplement), at least one vitamin or extract (eg plant extract) containing it.

Thus, according to a particular embodiment of the invention, the cosmetic method of the invention further comprises the simultaneous, sequential or delayed administration, by topical or oral route, of at least one vitamin or one extract in container.

It can alternatively include:

i. the application of a composition for topical application to keratin materials comprising an extract of a probiotic microorganism as defined in the invention preferably a Lactobacillus rhamnosus lysate (CNCM 1-5313), and at least one vitamin or one extract in container; or ii. the application of a first composition for topical application to keratin materials comprising an extract of a probiotic microorganism as defined in the invention preferably a Lactobacillus rhamnosus lysate (CNCM 1-5313), and administration by the oral of a second composition, preferably in the form of a food supplement, comprising at least one vitamin or an extract containing it; or iii. the application of a first composition for topical application to keratin materials comprising an extract of a probiotic microorganism as defined in the invention preferably a Lactobacillus rhamnosus lysate (CNCM 1-5313), and of a second composition topical comprising at least one vitamin or an extract containing it; or iv. the application of a first composition for topical application to keratin materials comprising an extract of a probiotic microorganism as defined in the invention preferably a lactate of Lactobacillus rhamnosus (CNCM 1-5313), and at least one vitamin or an extract containing it, and the administration of a second oral composition, preferably in the form of a food supplement, comprising at least one vitamin or an extract containing it.

By "simultaneous administration" is meant that the extract of probiotic microorganism according to the invention and the vitamin or vitamins are administered to the subject of interest, at the same time (same time).

By 'sequential administration' is meant that the extract of probiotic microorganism according to the invention and the vitamin or vitamins are administered to the subject of interest in a defined order, the vitamin or vitamins can be administered before or after said extract of probiotic microorganism according to the invention.

By "delayed administration" means that the probiotic microorganism extract according to the invention and the vitamin or vitamins are administered to the subject of interest in a defined order and at different times, more or less distant from each other depending on the desired effect.

The vitamin or vitamins may be water-soluble or liposoluble vitamins depending on the phase in which they are incorporated.

Among the water-soluble vitamins, there may be mentioned in particular:

- Group B vitamins: Bl (Thiamine), B2 (Riboflavin), B3 or vitamin PP (Niacin), B5 (Pantothenic Acid), B6 (Pyridoxine), B8 or Vitamin H (Biotin), B9 (Acid Lolic) and B12 (Cyanocobalamin),

- Vitamin C (Ascorbic Acid) and its derivatives such as ascorbyl glucoside, ethyl ascorbic acid, magnesium ascorbyl phosphate.

Among the fat-soluble vitamins, there may be mentioned in particular:

- Vitamin A (Retinol) and its derivatives such as palmitate and vitamin A propionate

- Vitamin E (alpha, beta, gamma, delta-tocopherol) and its acetate and phosphate derivatives as well as tocotrienol (alpha, beta, gamma, delta-tocotrienol) and its derivatives.

One can also use any known extract, in particular plant extract, for its richness in these vitamins.

The extract of a probiotic microorganism is generally present in a content ranging from 0.0001 to 10% by weight of dry matter, in particular from 0.0001 to 5%, preferably from 0.0001 to 2% , and in particular from 0.0001% to 1%, or even from 0.0001 to 0.05% by weight of dry matter relative to the total weight of the composition.

The cosmetic method of the invention is particularly intended for tired, aged, dull skin, impacted by an unfavorable or unbalanced nutritional context, emotional stress or lack of sleep.

The composition used in the cosmetic process can be a care and / or makeup composition, in particular a care composition for the skin of the face and / or of the body.

Composition and cosmetic set

Another object of the invention is a composition for topical application to keratin materials comprising, in a physiologically acceptable medium, at least one extract of a probiotic microorganism as defined in the invention, and at least one vitamin or a plant extract containing it, in particular a vitamin A, C, E and their mixtures.

According to a preferred embodiment, the extract of a probiotic microorganism according to the invention is a Lactobacillus rhamnosus lysate, preferably a Lactobacillus rhamnosus lysate (CNCM 1-5313).

By "composition" according to the invention means a cosmetic or dermatological composition.

According to a particular mode, it will be a cosmetic composition.

According to another particular mode, it will be a dermatological composition.

The composition according to the invention can, for example, be used as a care and / or makeup product for the face and / or the body.

The compositions according to the invention are intended more particularly for topical application to keratin materials, in particular to the skin.

The compositions of the invention comprise a physiologically acceptable medium, that is to say compatible with the skin and the integuments. The compositions may have any cosmetic form, and in particular may be in the form of an oil-in-water or water-in-oil emulsion or multiple emulsion, a microemulsion, a solution, a suspension, '' a lotion, gel, cream, milk, serum, mist (sprayed fluid solution), stick or powder and suitable for application on the skin, the lips and / or the integuments.

According to a particular embodiment, the composition of the invention is intended for topical application to the skin and / or the lips and is in the form of a cosmetic composition for caring for and / or making up the skin and / or lips.

The cosmetic composition of the invention is advantageously in the form of an emulsion, a serum, or a mist.

The composition generally comprises at least one aqueous phase and at least one oily phase.

The aqueous phase generally represents from 1 to 99% by weight, relative to the total weight of said composition.

An oily phase according to the invention may include hydrocarbon oils, silicone oils, fluorinated or not, and their mixtures. These oils can be volatile or non-volatile, vegetable, mineral or synthetic.

Advantageously, use will be made of hydrocarbon oils, in particular of vegetable origin.

As volatile hydrocarbon oils, mention may in particular be made of branched C8-C16 alkanes, branched esters of C8-C16 and mixtures thereof.

As non-volatile hydrocarbon oils, mention may in particular be made of hydrocarbon oils, hydrocarbon oils of plant origin, synthetic ethers of C10-C40, synthetic esters of C10-C40, fatty alcohols of C12-C26 , higher C12-C22 fatty acids, and mixtures thereof.

The oils may be present in the composition of the invention in a content ranging from 1 to 95% by weight relative to the total weight of the composition.

The composition of the invention may also include any additive usually used in cosmetics such as UV filters, antioxidants, surfactants, gelling agents, fillers, coloring matters (eg pigments), preservatives, film-forming polymers, perfumes, active cosmetic agents, such as, for example, emollients, moisturizers, vitamins, anti-aging agents, lightening agents, and mixtures thereof.

The invention also relates to a cosmetic assembly comprising at least a first composition for topical application comprising an extract of a probiotic microorganism as defined in the invention, preferably a lactate of Lactobacillus rhamnosus (CNCM 1-5313) , and at least a second composition for topical application or oral administration, preferably a food supplement, comprising a vitamin or a plant extract containing it, in particular a vitamin A, B, C, K, E and their mixtures.

The invention will now be illustrated in the following nonlimiting examples.

Unless otherwise indicated, the percentages are weight percentages expressed by weight relative to the total weight of the composition.

Examples

The Applicant evaluated the effect of a lysate of L. rhamnosus probiotic bacterial strain: ATCC 7469, CCM 1825, NCDO 243, NCIB 6375 or WDCM 00101 according to the collections in which it was deposited.

In particular, the probiotic bacterial strain used was deposited according to the Budapest Treaty with the Pasteur Institute (28 rue du Docteur Roux, F-75024 Paris cedex 15) on March 8, 2018 under the designation CNCM 1-5313 .

A lysate of this L. rhamnosus strain is obtained according to the following protocol from the supplier Greentech:

1. Culture of the L. rhamnosus strain in a suitable culture medium;

2. Biomass recovery by centrifugation;

3. Cell lysis (addition of soda);

4. Adjustment of the pH (addition of citric acid) and of the dry matter (addition of sterile water) to obtain an active ingredient content ranging from 0.01 to 10% by weight of dry matter (active ingredient) relative to the total weight of said lysate;

5. Possible addition of preservative (ex: phenoxyethanol).

Such a L. rhamnosus lysate is called Probiophyte LB by the supplier Greentech, and defined in INCI name under the name WATER and LACTOBACILLUS FERMENT LYSATE and PHENOXYETHANOL and SODIUM HYDROXIDE and CITRIC ACID, phenoxyethanol which can be replaced by another preservative .

This lysate comprises a content of Lactobacillus rhamnosus ranging from 0.1 to 10%, in particular from 0.5 to 5% by weight and in particular from 1 to 2%, or even 1.75% by weight of dry matter (active material) relative to the total weight of said lysate.

Such a lysate (but without phenoxyethanol preservative) is tested at a content of 0.1 and 0.2% in tests on cell cultures of skin cells and used in cosmetic compositions for topical application illustrated below at contents ranging from 1 to 2% by weight of raw material.

It is indifferently named LB Probiophyte or Lactobacillus rhamnosus lysate in the following examples.

Example 1: Pro-vitamin effect of the probiotic extract of L. rhamnosus on cultures of keratinocytes, melanocytes and fibroblasts

The test used uses a TLD A methodology (Taqman Low Density Array) which makes it possible to observe the stimulation or the inhibition or even the absence of stimulation of a large number of genes involved in skin processes of the skin. care of keratin materials, in particular on keratinocytes, fibroblasts and melanocytes.

It is a question here of evaluating the effect of the probiotic extract according to the invention on a targeted repertoire of genes corresponding to vitamin transporters and receptors.

[0108] Normal human keratinocytes (KHN), normal human fibroblasts (FHN) and normal human melanocytes (MHN) are cultured and then treated with the probiotic extract of L. rhamnosus described above.

After treatment of the KHNs, a study by Taqman Low Density Array (TLDA) on the genes studied is carried out using the cDNAs obtained after reverse transcription of the total RNAs extracted.

Cells used:

[0110] Cells: Normal human epidermal keratinocytes at the ^5th passage (KHN 12889), normal human fibroblasts of the dermis to the ^6th passage (HLF) and human normal melanocytes to ^7th passage (161 HLM) Test compounds:

Probiophyte LB without preservative: extract of Lactobacillus rhamnosus CNCM I5313

Doses tested: 0.1 and 0.2%

Processing time: 24h

N = 3 cultures by experimental conditions

Analyzes performed:

Measurement of gene expression by RT-qPCR from the messenger RNAs extracted from untreated cell mats (controls) and treated with the different concentrations of Probiophyte LB.

TLDA technology (TaaMan Low Density Array) • Obtaining total RNA

The cell culture medium is eliminated, and 250 μL of RLT lysis buffer (supplied in the Nucleospin RNA trace kit, Macherey-Nagel) are added. The cells are scraped using a Cell Scraper and the cell lysate is collected in a 1.2 mL deepwell (supplied in the Nucleospin RNA kit). Total RNAs are extracted according to defined protocols.

The total RNA solutions obtained are assayed, and their quality verified, using a microplate reader, spectrostarNANO (BMG Labtech) coupled to MicrolabSTAR. This device is connected to the computer controlling the Robotics platform and has specific software for analyzing the results (MARS software). The technique requires a 384-well micro-plate (LoBase), a positive control (RNA 250, AM7155, Thermofisher) to validate the pipetting performed by the robot as well as the values generated by the spectrostarNANO reader.

• Synthesis of complementary DNAs

The reverse transcription kit (RT) used is the High Capacity Reverse Transcription Kit (Thermo Lisher). It was used according to the protocol provided. 500 ng of total RNA are diluted in water for a final volume of 25 pL. They are then incubated for 10 minutes at 25 ° C and then 2 hours at 37 ° C in the presence of 25 μL of reaction mixture of High Capacity Reverse Transcription Kit 2X previously prepared as indicated below. The different incubations are done within TRobot (Biométra). [Tables 1]

Reagents RT buffer dNTP Primer RNase OUT RT H2O Volume 5 pL 2 pL 5 pL 0.5 pL 2.5 pL 10 pL

High Capacity Reverse Transcription Kit 2X reaction mixture for 1 reaction • PCR-TaqMan Low Density Array

15 μL of each RT are mixed with 60 μl of water and then 75 μL of TaqMan Gene Expression master mix (ThermoEisher), containing the DNA polymerase, are added. After homogenization, 100 μL are deposited on the microfluidic cards containing the probes corresponding to the genes tested, the latter are centrifuged and then sealed. The CD Rom corresponding to the profile of the genes deposited on the plates is loaded into the SDS 2.3 software, specifying the location of each gene on the map. The control gene (or “endogeneous gene”) to be used for the normalization of the results is to be indicated before the launch of the PCR. The latter is carried out according to the protocol provided by Applied Biosystems in the ABI Prism 7900HT Sequence detection system. The qPCR steps are 2 min at 50 ° C, 10 min at 94.5 ° C then 30 s at 97 ° C and 1 min at 59.7 ° C for 40 cycles.

Statistical analyzes

The quantitative real-time PCR can be exploited if its efficiency is between 90% and 110%. For each sample, the number of cycles at which the signal appears is determined by the SDS 2.3 software. For the same test, the expression levels of the transcripts of interest obtained are normalized with respect to the value obtained for the household gene Beta-2-microglobulin. This gene, the expression of which is constitutive and invariant, makes it possible to overcome any variations induced during the experiment (assay of total RNAs, pipetting, reverse transcription step, PCR in the apparatus).

In the TLDA RT-PCR method, the quantification is carried out using the AACt comparative method. The relative quantization values (RQ) obtained correspond to the level of amplitude (x times more or less than the control) of the expression compared to our control here the non-irradiated. The RQ is obtained by the following calculation where the control is equal to 1:

[0125] [Math.l]

RQ = 2-ΔΔ (Ξΐ = 2- (ACt treated - ΔΟΐ not treated)

[0126] ACt treated = Ct target gene treated - Ct household gene treated

Untreated ACt Ct Untreated Target Gene - Ct Untreated Household Gene

In order to assess variations in statistically significant transcriptional activity, we will use Student's t test. Each condition is performed in triplicate (3 untreated and 3 treated under the same conditions). Eischer's E-test is first applied by comparing the two data matrices. When the value is greater than a = 0.05 then the variance for Student's t-test is 2, when the Eischer's E-test is less than a = 0.05 then the variance will be equal to 3. Transcriptional variations retained will be those which have a Student's t test less than a = 0.05.

The results are presented on average over n = 3. Student's t-test was used to compare the effect between treated and untreated cells.

The results are considered to be significant for p <0.05 (*) or p <0.01 (**).

The results presented in Table 2 below show the stimulating effect of the LB probiophyte on several genes of vitamin receptors and transporters in cultures of keratinocytes, melanocytes and fibroblasts.

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Fibroblasts Keratinocyte s Melanocytes Category es of genes Genoa Symbols Refer this NCBI Probi ophyt eLB 0.1% Probi ophyt eLB 0.2% Probi ophyt eLB 0.1% Probi ophyt eLB 0.2% Probi ophyt eLB 0.1% Probi ophyt eLB 0.2% Vitamin receptors Cubilin CUBN 8029 2.128 2,321 - - - - Folate receptor 3 F0LR3 2352 - - - 1,380 - - Hydroxycarb oxylic acid receptor 2 HCAR 2 338442 2,314 2.927 - - - - Hydroxycarb oxylic acid receptor 3 HCAR 3 8843 - 3,497 - - - - Retinol binding protein 1 RBP1 5947 - - 1,273 - - 4,112 Scavenger receptor class B member 1 SCAR Bl 949 - - 1.239 - - - Stimulated by retinoic acid STRA6 64220 3,288 2,591 - - - - Transcobala min 1 TCN1 6947 - - 1,764 2.309 - - Transcobala min 2 TCN2 6948 2.502 2.406 - - - - Vitamin transporters ATP binding cassette subfamily A ABCA 1 19 3,631 3.962 1,258 - - -

member 1 Solute carrier family 19 member 3 SLC19 A3 80704 - - 1,490 - 1.652 1,924 Solute carrier family 2 member 3 SLC2A 3 6515 - - 1,320 1,500 - - Solute carrier family 52 member 1 SLC52 Al 55065 - 3,488 - - - -

These results show that all cell types are affected by the stimulating effects of the LB Probiophyte on vitamin transporters and receptors with, however, more reduced stimulation for melanocytes.

The vitamins concerned by stimulation by the Probiophyte LB, of the genes of their transporters are:

[0134] - vitamins K (ABCA1 / LHN and KHN)

[0135] - Vitamin B1 (SLC19A3 / KHN and MHN)

[0136] - Vitamin C or more exactly dehydroascorbic acid (SLC2A3 / KHN):

note that this transporter is better known for transporting glucose and other monosaccharides, galactose, mannose, xylose and fucose, and

[0137] - vitamin B2 (SLC52A1 / LHN),

The vitamins concerned by stimulation by the LB Probiophyte, of the genes of their receptors are:

[0139] - Vitamin B12 (CUBN / LHN; TCN1 / KHN and TNC LHN)

- Vitamin B9 (LOLR3 / KHN)

[0141] - vitamin B3 or niacin (HCAR2 and 3 / LHN),

[0142] - vitamin A / retinol (RBP1 / KHN, MHN; STRA6 / LHN),

[0143] - vitamins E and K (SCARB1 / KHN)

All of these results show that the extract of a microorganism according to the invention allows better bioavailability in the skin of the vitamins concerned and their use by skin cells via (i) activation of transporters vitamins (ABCA1, SLC19A3, SLC2A3 and SLC52A1), and facilitates the biological action of these vitamins on skin cells via (ii) activation of vitamin 17 receptors.

Example 2: Cosmetic formulations for topical application

The following formulations are prepared according to the conventional methods of formulations in the cosmetic field.

Anti-fatigue cream

[0147] Triclyceride of caprylic / capric acids 10%

7% glycerin

[0149] Cetearyl alcohol 3%

2% isostearyl hydroxy stearate

Isotridecyl isononanoate 2%

[0152] Steareth-21: 1%

1% sodium polyacrylate

0.5% xanthan gum

0.5% phenoxyethanol

0.5% sodium hyaluronate

Tocopherol acetate (vitamin E) 0.05%

[0158] L. rhamnosus lysate 1.5%

0.1% Vitamin A Palmitate

Ascorbyl glucoside (vitamin CG) 2%

Water qs 100%

Applied to the skin, the composition makes it possible to impart a moisturizing effect, good looks, and a radiant complexion.

Vitamin serum

4% glycerol

[0164] Butylene glycol 3%

Carbomer 0.5%

1% lactic acid

[0167] Lysate of L. rhamnosus 2%

0.7% sodium hydroxide

0.1% xanthan gum

0.2% acrylic gelling agent

Alcohol 3%

[0172] Ascorbyl glucoside (vitamin CG) 0.5%

0.5% phenoxyethanol

Dyes qs%

Eauqspl00%

Applied to the skin, the composition makes it possible to improve the firmness of the skin and give it a healthy glow effect.

Mist

2% glycerol

4% glycols

2% gelling agents

Lysate of L. rhamnosus 1%

0.25% caprylic / capric acid triclyceride

Emulsifiers 0.6%

0.5% phenoxyethanol

Tocopherol (vitamin E) 0.02%

Water qs 100%

Applied to the skin, the composition makes it possible to promote and / or improve the nutrition of keratin materials and give it a healthy glow effect, a radiant and homogeneous complexion.

In a particular mode, the topical formulations illustrated above can be combined with an oral administration of vitamin C or commercial multivitamin cocktail.

Claims (1)

Claims [Claim 1] Use of at least one extract of a probiotic microorganism, one of its fractions and / or one of its metabolites, in a composition for topical application to keratin materials, as a provitamin agent, capable in particular of promoting and / or improving the nutrition of keratin materials and / or vitamin efficiency. [Claim 2] Use according to claim 1, characterized in that the extract of a probiotic microorganism is a lysate of probiotic microorganism, of one of its fractions and / or of one of its metabolites. [Claim 3] Use according to claim 1 or claim 2, characterized in that the probiotic microorganism belongs to the genus Lactobacillus, to the genus Bifidobacterium, or to their mixture, preferably to the genus Lactobacillus chosen from the group consisting of: Lactobacillus acidophilus, Lactobacillus paracasei, Lactobacillus casei , Lactobacillus rhamnosus, Lactobacillus thermophylus, and Lactobacillus johnsonii. [Claim 4] Use according to claim 3, characterized in that the probiotic microorganism belongs to the species Lactobacillus rhamnosus, in particular the strain Lactobacillus rhamnosus (CNCM 1-5313). [Claim 5] Non-therapeutic cosmetic method intended to promote and / or improve the nutrition of keratin materials, comprising the application to keratin materials, in particular the skin and / or its integuments and / or the lips, of at least one extract of a probiotic microorganism as defined in any one of claims 1 to 4, preferably a lactate of Lactobacillus rhamnosus (CNCM 1-5313), or of a cosmetic composition for topical application containing it. [Claim 6] Cosmetic method according to claim 5, characterized in that it further comprises the simultaneous, sequential or delayed administration, by topical or oral route, of at least one vitamin or an extract containing it. [Claim 7] Cosmetic method according to either claim 5 or claim 6, characterized in that the extract of a probiotic microorganism is present in a content ranging from 0.0001 to 10%, in particular from 0.0001 to 5%, in particular from 0.0001 to 2% by weight, and in particular from 0.0001% to 1%, or even from 0.0001 to 0.05% by weight of active material (dry matter) relative to the total weight of the composition. [Claim 8] Cosmetic method according to any one of Claims 5 to 7, characterized in that it is intended in particular for tired, aged, dull skin, impacted by an unfavorable or unbalanced nutritional context, emotional stress or lack of sleep. [Claim 9] Composition for topical application to keratin materials comprising, in a physiologically acceptable medium, at least one extract of a probiotic microorganism as defined in any one of claims 1 to 4, preferably a lactate of Lactobacillus rhamnosus (CNCM 1- 5313), and at least one vitamin or an extract containing it, in particular a vitamin A, B, C, K, E and their mixtures. [Claim 10] Cosmetic assembly comprising at least a first composition for topical application comprising an extract of a probiotic microorganism as defined in any one of claims 1 to 4, preferably a lactate of Lactobacillus rhamnosus (CNCM 1-5313), and at least a second composition for topical application or oral administration, preferably a food supplement, comprising a vitamin or a plant extract containing it, in particular a vitamin A, B, C, K, E and their mixtures. FRENCH REPUBLIC